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Deliverable 2.5: TECHNICAL DATASHEETS IN INNOVATIVE SOLUTIONS TO CONTROL

GRAPEVINE TRUNK DISEASE AND FLAVESCENCE DORÉE
INSTITUTO GALEGO DE CALIDADE ALIMENTARIA- INGACAL-
MARÍA JESÚS FANJUL
02/05/2017
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Network for the exchange and transfer of innovative
knowledge between European wine-growing regions to
increase the productivity and sustainability of the sector
This project has received funding from the European Union’s Horizon 2020 research and
innovation programme under grant agreement No 652601
DELIVERABLE 2.5– VERSION 1
WORK PACKAGE N° 2
Nature of the deliverable
R Document, report (excluding the periodic and final reports) X

DEM Demonstrator, pilot, prototype, plan designs
DEC Websites, patents filing, press & media actions, videos, etc.
OTHER Software, technical diagram, etc.
Dissemination Level
PU Public, fully open, e.g. web X

CO Confidential, restricted under conditions set out in Model Grant Agreement
CI Classified, information as referred to in Commission Decision 2001/844/EC
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ACKNOWLEDGMENTS
This report forms is part of the deliverables from a project called "Winetwork" which has received
funding from the European Union’s Horizon 2020 research and innovation programme under grant
agreement No 652601.
Winetwork project aims at to create a Network for the exchange and transfer of innovative
knowledge between European wine-growing regions to increase productivity and sustainability of
the sector. For 30 months, 11 partners of 7 European countries, representing more than 90% of the
EU wine production, will exchange on their knowledge on two important diseases in vineyard: the
Grapevine Trunk Diseases and Flavesence Dorée. These diseases are well-known in many vineyards
and have been extending for several years in different European countries. This network will
promote interactions between scientists and practitioners to gather and share experiences and
knowledge of different actors from the main wine producing European regions.
More information on the project can be found at http://www.winetwork.eu.
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TABLE OF CONTENTS
1. INTRODUCTION ............................................................................................................................... 5
2. METHODOLOGY ............................................................................................................................... 5
2.1. Data sheet template definition .................................................................................................... 5
2.2. Topic Definition ............................................................................................................................ 6
2.3. Working Procedure. ..................................................................................................................... 7
3. RESULTS ........................................................................................................................................... 8
3.1. Technical data sheets on Good Practices ..................................................................................... 8
3.2. Specific Data Sheets ..................................................................................................................... 9
4. CONCLUSION ................................................................................................................................. 10
5. APPENDICES ................................................................................................................................... 11
5.1. APPENDIX 1: GOOD PRUNING PRACTICES .................................................................................. 11
5.2. APPENDIX 2: GLOBAL VINEYARD STRATEGY TO PREVENT GTDs INFECTION .............................. 11
5.3. APPENDIX 3: TRICHODERMA APPLICATION FOR PROTECTING GRAPEVINE PRUNING WOUNDS

11
5.4. APPENDIX 4: A GUIDE OF GOOD PRACTICES FOR REGIONS WITHOUT FLAVESCENCE DORÉE ... 11
5.5. APPENDIX 5: FLAVESCENCE DORÉE: HOW TO MANAGE THE DISEASE WITH MORE PRECISION 11
5.6. APPENDIX 6: TRUNK RENEWAL .................................................................................................. 11
5.7. APPENDIX 7: INTRODUCTION OF SMALL WOOD STICKS INOCULATED WITH TRICHODERMA ... 11
5.8. APPENDIX 8: PRUNING WITH REGARD TO SAP FLUX ................................................................. 11
5.9. APPENDIX 9: TRUNK CLEANING .................................................................................................. 11
5.10. APPENDIX 10: PEROXYDE HYDROGEN INJECTION .................................................................... 11
5.11. APPENDIX 11: USE OF MYCORRHIZAE IN FIELD........................................................................ 11
5.12. APPENDIX 12: COPPER NANOPARTICULES INJECTIONS ........................................................... 11
5.13. APPENDIX 13: ORANGE OIL APPLICATION ............................................................................... 11
5.14. APPENDIX 14: HOT WATER TREATMENT ................................................................................. 11
5.15. APPENDIX 15: WILD VINES MANAGEMENT ............................................................................. 11
5.16. APPENDIX 16: MATING DISRUPTION........................................................................................ 11
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1. INTRODUCTION
One of the aims of this Project is to synthesize practical and scientific knowledge about all means and
practices used for controlling and management of Grapevine Trunk Diseases and Flavescence Dorée
in winegrowing regions within Winetwork project. The final goal is to spread, in an adapted way,
among specific target groups (technicians, winegrowers and institutions involved in innovation
transfer and support) relevant information obtained in order to control and mitigate serious
damages produced by these 2 diseases in European vineyards.
The synthesis of this knowledge is reflected in some Technical Datasheet. Those involve as much
scientific, technical and practical information as possible about means and practices observed in
regions within Winetwork project
Therefore, the goal of this deliverable is to comprise the content of Technical Datasheets elaborated
by Facilitator Agents in the 10 regions involved in Winetwork project, which synthesize this scientific-
practical knowledge and will be the basis to elaborate material to be disseminated.
In this sense, technical sheets about good practices have been elaborated for management and
control of EMV and FD. They can either be innovative or not, can have a prophylactic or preventive
character and count on a scientific basis that which can be verified before its implementation.
Moreover, they have collected some other datasheets containing innovative solutions carried in
some European regions, which in some cases do not count on verifiable scientific bases , but whose
results are promising for the winegrowers applying them and can be interesting for winegrowers in
other region where they are unknown and also as study data or backing for investigation
2. METHODOLOGY
Every FA in their regions has worked in a coordinated and homogeneous way on the basis of two
pillars: a Data Sheet template and a working procedure previously agreed between all FA in the
meeting held on the 15th-16th December 2016 in DLR Neustadt and. Weinstraße.
2.1. Data sheet template definition
A model template with potential contents was initially forwarded to all FA for their analysis and
improvements proposals. Later on, in a 2nd meeting held by the FA in Germany, those contents were
discussed and they jointly agreed its structure and main sections to be included, as they are defined
here after.
Practice Description
Application area (when relevant, not relevant in the case that the practice is widely used)
Scientific data with links to scientific publication
Background
Outcomes of the practices
Key points for success and risks (conditions of success, benefits/disadvantages)
Innovative aspects (why is it innovative?)
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Other requirements (adaptability, training needed, costs, regulation, …)
Specific cases (illustrate the practice with a concrete case of a winegrower)
More information
Bibliography
2.2. Topic Definition
In order to define the main topic to be covered in data sheets it has been taken into account:
- All scientific and practical knowledge about GTDs and FD gathered in former stages of the project
by both FA and SWG members.
- For GTDs control, given the wealth of practices observed in the field, it has been taken into
account previous evaluation by SWG about which practices are scientifically backed and which of
them would require more information and investigation for its efficiency to be proved.
- Analysis with TWG members of every region about practices detected both in their own region
and in other regions within the project which can be interesting for their region
Therefore, after a common agreement reached in their meeting in Germany, the 10 FA decided to
reflect all the knowledge to that moment in the following main topics:.
5 subjects gathering Good Practices for GTD and FD control and management on themes
indicated in Table 1.
They are preventive practices, innovative or not and with verifiable scientific results
Table 1. Subject of Data Sheet on Good Practices
TOPICS OF TECHNICAL DATA SHEET
Good pruning practices

Grapevine
Trunk Global vineyard strategy to prevent GTDs infection
Disease
Trichoderma application by spraying
For the regions where FD is not present yet: Presentation of the

Flavescence disease and its risks
Dorée
For the regions where FD is already present: how to improve FD
management
Apart from these themes, there are some other innovative practices specific of one or more
regions. They are considered relevant by FA and TWGs in their regions and even thought
most of them do not have verifiable scientific data available, they have shown satisfactory
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results for the winegrowers applying them. Table 2 shows selected practices of which
technical data sheets will also be available.
Table 2. TOPICS of Technical Data Sheet on specific region practices
Type of Disease PARTICULAR TECHNICAL DATA SHEET
Trunk renewal
Introduction of small wood sticks
inoculated with Trichoderma
Pruning with regard sap flux

Grapevine Trunk
Disease Trunk cleaning
Peroxide Hydrogen injection
Use of mycorrhizae in field
Copper nanoparticules injections

Orange oil application
Hot water treatment
Flavescence Dorée Wild vines management
Mating disruption
2.3. Working Procedure.
a) Division of topics in groups :
The Facilitator agents were divided into groups according to the topics to be worked in each
Technical Datasheet.
In relation to the preventive practices which are of common interest for all regions, three
working groups were organized for GTDs and two for FD depending in the latter, there are
regions with this disease present and others not.
In the event of innovative or atypical practices, some of which, have not been scientifically
validated, some working groups were created, participating in every group the AF of the
regions where the practices had been considered relevant. The FA have also worked together
in order to supply all available information about those practices, specifying those aspects
which are distinctive of the region where they were detected
b) Appointing a coordinator: each group has a coordinator responsible for organizing tasks
provided by each member, unifying and giving a final format to technical documents They
will also transfer the results to project and specific task coordinators.
c) Revision by SWG : Once Technical Data Sheets were finished , they were sent to SWG
members for revision of their contents.
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d) Development of a final version. This version will serve as the basis for elaborating the end-
use flyers. Moreover, it will also be part of the Knowledge Reservoir in the project’s web in
the languages mentioned in the following section.
3. RESULTS
Topics referred to in point 2, have generated to kind of technical data sheets : those collecting good
practices and those collecting specific practices
3.1. Technical data sheets on Good Practices
40 Technical data sheets have been elaborated on Good Practices for control and managements on
GTDs and FD, whose content is presented in appendixes 1 to 5 of this document.
24 regarding GTD. Correspond to the elaboration of the 3 main topics for control and
management of GTDS in English language and the languages of the 7 regions within
Winetwork Project.
16 Regarding FD. Correspond to the elaboration of the 2 main topics for control and
management of FD in English language and the languages of the 7 regions within Winetwork
Project.
Table 3 Data Sheet on Good Practices
Languages of Technical Data Sheet

Subject of Technical Data Sheet
English-Spanish-Portuguese-Italian-German-Croatian-Hungarian
Good pruning Practices
Grapevine Trunk Global vineyard strategy to prevent GTDs infection

Disease
Trichoderma application by spraying
Regions where FD is not present yet: Presentation of

the disease and its risks
Flavescence Dorée
Regions where FD is already present: how to improve FD
management?
These Technical Data Sheets provide the knowledge basis for designing and elaborating end- user
flyers on each region.
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3.2. Specific Data Sheets
25 Specific Data Sheets were elaborated (16 for GTDs and 9 for FD), produced in the languages
of the regions specified in Tables 4-5 and also in English when more than one region is involved.
Its content is gathered in appendixes 6 to 16 in this document
Since some of these Data Sheets are not scientifically validated, they will not be used for its
dissemination by end-users flyers. However, they can be an important documentary source for
technicians, members of the TWGs and ISS, providing information on practices that might be of
interest to the region or on areas that require further research to verify their feasibility. Those
practices will be collected in a booklet about innovative and atypical practices detected during
the project.
Table 4 Data Sheet on specific practices to manage GTD

English-French-Spanish-Portuguese-Italian-German-Croatian-Hungarian
Trunk renewal
Introduction of small wood sticks
inoculated with Trichoderma
Pruning in regard with sap flux
Trunk cleaning
Peroxyde Hydrogen injection

Use of mycorrhizae in field
Copper nanoparticules injections
Table 5 Data Sheet on specific practices to FD

English-French-Spanish-Portuguese-Italian-German-Croatian-Hungarian
Orange oil application
Mating disruption
Hot water treatment
Wild vines management
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4. CONCLUSION
Through technical data sheets we have managed to collect in the same documents the most updated
scientific knowledge and practical experience.
The actions taken by the participants in the Project (TWG, FA and SGW) plus the interactions within
every region and between the different regions has contributed to improve the knowledge about
these diseases and to generate accessible documentation for specific target groups.
This documentation could be used for its diffusion in the Knowledge Reservoir and through the
disclosure actions organized by the members according to their dissemination plans.
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5. APPENDICES
5.1. APPENDIX 1: GOOD PRUNING PRACTICES

5.2. APPENDIX 2: GLOBAL VINEYARD STRATEGY TO PREVENT GTDs INFECTION
5.3. APPENDIX 3: TRICHODERMA APPLICATION FOR PROTECTING GRAPEVINE
PRUNING WOUNDS
5.4. APPENDIX 4: A GUIDE OF GOOD PRACTICES FOR REGIONS WITHOUT
FLAVESCENCE DORÉE
5.5. APPENDIX 5: FLAVESCENCE DORÉE: HOW TO MANAGE THE DISEASE WITH MORE
PRECISION
5.6. APPENDIX 6: TRUNK RENEWAL
5.7. APPENDIX 7: INTRODUCTION OF SMALL WOOD STICKS INOCULATED WITH
TRICHODERMA
5.8. APPENDIX 8: PRUNING WITH REGARD TO SAP FLUX
5.9. APPENDIX 9: TRUNK CLEANING
5.10. APPENDIX 10: PEROXYDE HYDROGEN INJECTION
5.11. APPENDIX 11: USE OF MYCORRHIZAE IN FIELD
5.12. APPENDIX 12: COPPER NANOPARTICULES INJECTIONS
5.13. APPENDIX 13: ORANGE OIL APPLICATION
5.14. APPENDIX 14: HOT WATER TREATMENT
5.15. APPENDIX 15: WILD VINES MANAGEMENT
5.16. APPENDIX 16: MATING DISRUPTION
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APPENDIX 1: GOOD PRUNING PRACTICES
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TECHNICAL DATA SHEET
GOOD PRUNING PRACTICES
Pruning wounds represent a significant point of entry to grapevine trunk diseases fungi, while pruning debris and
symptomatic vines are a source of fungal inoculum. Implementation of preventative control strategies needs to be
adopted early after vineyard establishment but winegrowers mainly start to conduct control strategies after grape-
vine trunk diseases’ leaf symptoms appearance.

knowledge between European wine growing regions
This project has received funding from the European Union’s Horizon 2020
research and innovation programme under grant agreement No 652601
depending on the training system, corresponding to 15-20%

Factors related to pruning on Guyot double, 10-25% on Guyot simple, 0-5% gobelet
that may influence grapevine or Robat, and 0-1% on cordon training system. Moreover,
changes in cultural practices in Tuscany, such as replacement
trunk diseases development of cordon with Guyot training system, led to an increase of
Esca disease. A correlation of Esca leaf symptoms incidence
Factors related to pruning such as training system, weather with cane length on Guyot training system was evaluated
condition during pruning period, number and size of pruning in the winegrowing area of Bordeaux, results indicate that
wounds, location and aggregation of pruning wounds, cane symptoms incidence was higher on Guyot-trained
and spur length, wound protection, wound age, late seaso- vines with shorter cane length. Development of Eutypa die-
nal pruning, pruning debris management contribute to the back foliar symptoms is higher on spur pruned vines in com-
incidence of wound infections and grapevine trunk diseases parison to cane pruned, but lower death rate than on cane
(GTD) development. pruned vines (Fig. 1). On Fig. 1 can be observed that cane
Impact of training system on GTD incidence and se- pruned vines have numerous grouped wounds on upper part
verity has been observed in different winegrowing regions of grapevine trunk, while spur pruned vines have a greater
but the available information is partial or contradic- total wound surface.
tory. Some authors state that different training systems and
pruning methods increase the risk of inner wood necrosis Pruning wounds represent a point of entry to vascular grape-
development and perennial wood infection with fungi related vine pathogens, such as fungi implicated in GTD, which are
to these diseases, but the research was conducted in diffe- able to overcome grapevine defence mechanism due to their
rent vineyards and these conditions would have an impact on virulence characteristics. Large and numerous pruning
research results. Differences in Esca incidence are observed wounds, usually frequent on older vines or vines that had
Figure 1: Correlation of grapevine training systems with GTD infection. (Sosnowski, 2016)
been retrained in a different training system, provide an lity is higher when vines are pruned early in dormant season
important point of entry to fungi due to greater total and lower when are pruned later in the dormant season.
wound surface area where spores can land and induce in-
fection. Esca complex, a GTD caused by numerous fungi that be-
Spread pattern of GTD fungi in vineyard is connected with long to different taxonomical classification, has a lifecycle
the distribution of infected vines, where newly symptomatic that differs depending on the fungi species present within
vines are usually located close to previously infected vines. the vineyard. Spore release of Phaeomoniella chlamydospora
Some fungal inoculum is transmitted to pruning wounds is correlated to rainfall, while for Phaeoacremonium mi-
with pruning shears from infected vines, but the inoculum nimum occurs during the vegetative period without any link
concentration transmitted with pruning shears is insigni- to rainfall. The infection of pruning wounds by Pa. chlamy-
ficant. dospora decreased from 75% to 10% when inoculation oc-
curred 12 weeks after pruning.
Pruning period: wheather
condition Pruning grapevines during dry weather is critical because
fungal airborne inoculum is significantly lower at that period.
Late pruning in the dormant season (as close as possible to
To choose the most appropriate pruning period it is necessa- bud break) is a recommended cultural practice since pruning
ry to consider different factors such as: specific climatic wounds heal faster with high degree-day temperatures.
conditions in the concerned winegrowing region, different Recent studies indicate that the rate of natural infection of
life cycles of GTD pathogens, spore release and wound pruning wounds is lower following early pruning (autumn)
infection susceptibility depending on the weather conditions, than following late pruning (winter). The susceptibility of the
pathogen virulence. wounds is mainly influenced by the relative humidity and
rainfall periods.
It has been reported that incidence and type of symptoms
of different GTD vary greatly between regions. This indicates
that rainfall and temperature influence not only the
distribution of pathogens but also the symptoma-
tology of the pathogens in a climatic region. Furthermore, Weather conditions are significant for release
it was observed that the pathogens overlapped in terms of and dispersal of fungal species implied in grape-
symptomatology, making symptom-based diagnosis of these vine trunk diseases, therefore pruning should be
diseases and their causal organisms unreliable. Therefore, conducted during dry periods.
management strategies for the different pathogens in a spe-
cific region should be aimed at the whole complex of trunk
disease pathogens.
Botryosphaeria dieback, a GTD caused by numerous

fungi belonging to Botryosphaeriaceae family, is spread wit-
hin the vineyard by airborne inoculum, especially during
rainfall or during overhead sprinkler irrigation. Aerial inocu-
lum was observed during winter in California, while it was
mainly detected during the vegetative period in France. Be-
cause of this, in California, the wound susceptibility is higher
when vines are pruned in dormant season and lower when
vines are pruned in early March. On the contrary, in France, it
was found that the wound susceptibility is higher after blee-
ding (mean temperature > 10°C).
Eutypa dieback, a GTD caused mainly by E. lata, is fre-

quently found in vineyards that receive more than 250 mm
of rainfall per year, due to spore release throughout the entire
year and spore dissemination with each rainfall > 0.5 mm.
Spores are released in the frame of 2-3 h after the onset of
rain and stops 24 h after the rain stops. The fungi penetrates
into the plant through pruning wounds (spore germinate IFV Alsace
into the wound), and it was found that the wound susceptibi-
Management of pruning Protection of pruning

debris and other sources of wounds
fungal inoculum
The source of inoculum of GTD fungi can be found on vines Adoption of preventative control methods oriented to GTD
that display wood and/or foliar symptoms and other crops management early after vineyard establishment is critical.
like orchards cultivated near vineyards. Fungal inoculum Infection rate in a longer period of time is significantly lower
can be found on necrotic stems, leaves, desiccated if a control strategy, with control efficiency of 75% pruning
bunches, under the bark of perennial wood (trunk, wounds, is regularly applied 3 to 5 years after vineyard es-
cordon), dead wood and pruning debris of grapevine and tablishment. Results presented in Fig. 2 indicate potential
it represents a potential source of new infections in vineyard. efficiency of practices such as late pruning, double pruning
In order to remove the source of infection different practices and pruning wound protection (mastic or spray application).
are applied in European vineyards, the most common applied Preventive disease management, if implemented early after
by winegrowers are mechanical grinding followed by incor- vineyard establishment will minimize disease development
poration in soil, burning, mechanical grinding followed by and additional costs of cultural practices like trunk renewal
composting, extirpation of symptomatic and/or dead vines. or replacing vines later in production.
Often, there are concerns on the impact of those practices in
fungal eradication and prevention of fungal dispersal.
It is estimated that pruning debris is a potential source

of Botryosphaeria dieback fungi for 42 months, but
infective inoculum decreased significantly after 24 months
and spores viability was reduced to 44%.
Pruning debris and other grapevine fragments may be rein-
troduced in vineyard after a process of mechanical grinding
and composting since this procedures eliminate the GTD fun-
gi, if applied adequately, and do not pose a risk of vineyard
recontamination with Eutypa dieback, Esca or Botryosphae-
ria dieback.
Mechanical grinding and composting on 40 – 50°C

for a period of six months eradicated successfully GTD
fungi (compost made from 140 m3 pruned and grounded
vine material, 125 m3 sheep manure, 60 m3 of stalks and
garden residues like grass and leaves). Additionally, some
GTD fungi that induce Esca disease (Pa. chlamydospora
and P. aleophilum) were not isolated from grapevine
fragments after grinding, the authors assume that grin-
ding favoured the activity of saprophyte fungi that grow more
rapidly than those GTD species, but precise scientific data
that confirms this finding is still missing.
Fig. 2 A) Infection rate with no action and 75% disease control effi-
ciency. B) Yield per acre for healthy, no action implemented, and 75%
disease control efficiency (Baumgartner et al., 2014).
It is significant to remember that wounds remain potential wound protection is achieved with sprayed formulations
infection pathways to fungi for a long period of time and or paintbrush applications of fungicides. Sprayed for-
protection of both new and old pruning wounds is re- mulations are more practical, time and cost effective but they
quired to limit disease establishment on yearly basis. Pruning are easily washed off by rainfall.
wound protection is oriented to biological or chemical fun-
gicides, both applied as prevention, posing different critical Biological control agents (e.g. Trichoderma spp.) and
points to consider in disease management. natural molecules (e.g. chitosan) have been reported
efficient in pruning wound protection, in addition biologi-
cal agents (BCA) are able to actively colonize pruning
As a preventive measure applied in grapevine trunk
wounds up to 8 months. Treatments with BCA 6 hours
diseases management in vineyards, protection of
after pruning, either early or late pruning, resulted in high
pruning wounds is an essential step.
wound colonization with Trichoderma spp. even if weather
conditions and physiological state of grapevine were diffe-
rent at that vegetative state.
One of the major limits of chemical fungicides is the nar- Once the disease is established it is difficult to implement
row period of residual activity. Pruning is usually conducted a successful eradication as a result of limited efficiency of
early in the season since organization of work and weather available control strategies. The development of some GTD
conditions require a longer period of time and it is unlikely to can have two forms – chronic and apoplectic. Consequently,
achieve efficiency that lasts from 1 to 2 months. Some che- even if infections primarily occur through pruning wounds on
mical fungicides have been reported to be efficient even after upper parts of grapevine, GTD fungi progress overtime leads
3 weeks post treatment application and when needed more to colonization of more distant perennial parts of grapevine,
than one treatment may be applied. Application of pruning such as trunk (Fig. 3).
Figure 3: Potential progression of GTD fungi from cordon into basal parts of the trunk.
(1-3: symptomatic wood, 4: asymptomatic wood), (Sosnowski, 2016).
sap route from one year to another with pruning that po-
Innovative/alternative sitions wounds only on the upper part of the cordon (Fig. 5).
aspects Guyot-Poussard pruning contributes to the small size and
low number of pruning wounds. Some pruning systems re-
1- Pruning Guyot-Poussard quire retraining and a return cut, common in older vineyards,
which could be avoided with this method of pruning. Mo-
The diameter of the pruning wound may be correlated with reover, wounds on older wood, common on retrained vines,
a 1.5 times longer necrosis on pruned spur or cane located are reported to be less resistant to GTD fungi infection than
near perennial wood (Fig. 4). Large wounds near perennial wounds on 1-year old wood. Impact of Guyot-Poussard
parts of grapevine cordon and/or trunk induce wood necrosis on reduction of GTD incidence and severity needs
that potentially lead to higher infection rates and dete- to be scientifically validated and current information is
rioration of sap flux. Moreover, deterioration of grapevine based only on hypothesis.
sap flux increases disease negative impact due to higher
stress in grapevine physiology. Symptoms incidence and se- 2- Double pruning
verity is increased, and apoplectic forms may be more fre-
quent on those vines. Double pruning is a modification of late pruning and recent-
ly it has been implemented in preventative management of
GTD in spur-pruned vineyards. This practice is not applicable
in cane pruned vineyards, but in spur pruned vineyards is an
efficient practice to delay pruning until March and reduce
infection rate of GTD pathogens.
Double pruning involves two operations that may be di-
vided into pre-pruning and pruning. Pre-pruning consists of
non-selective mechanical pruning, on uniform height
of approximately 30 - 45 cm above spur, a second cut to
form the desired pruning system is conducted later in
the season, usually as close as possible to bud brake (Fig. 6).
Pruning techniques that preserve longer length of two-year
Figure 4: Correlation between pruning wounds and necrosis develop- old cane above the upper winter bud reduce the infection of
ment. (Crespy, 2006)
grapevine perennial wood located on cordon and/or trunk,
due to limited yearly progress of GTD fungi. An economic
Pruning in regard with sap flux was adopted by Lafon (1927) evaluation estimates double pruning as highly cost practice
from a pruning system used in France, and later named in comparison with late pruning, while the efficiency of both
Guyot-Poussard according to its developer. The main prin- practices is similar.
ciple of this pruning system is maintenance of the same
Figure 5: Grapevine pruning system Guyot-Poussard. (http://simonitesirch.com)

{ {
Figure 6: Mechanical pre-pruning (left), manual spur pruning (right) (IFV South-West)
3- Minimal pruning
Minimal pruning consists of almost no pruning and recently it

has been considered as a cultural practice with a potential to
reduce the infection rate of pruning wounds with GTD fungi.
While this system reduces labour costs of pruning, it is also
related with high productions and lower grape quality.
Minimally pruned vines, in comparison with spur-pruned
vines, have lower: wood necrosis, Esca disease incidence
(leaf symptoms), variability in fungal community and inci-
dence of virulent fungal trunk pathogens. A research related
to the impact of pruning systems on Eutypa dieback indi-
cated that disease incidence and severity are lower on mi-
nimally pruned vines when compared to spur pruned vines.
Minimal pruning- IFV South-West
Summary - Critical points • Minimize the number of wounds on grapevine in

general (damage due to mechanical harvest, mechani-
cal sucker removal, mechanical pruning, freeze damage,
Reduce fungal infective inoculum etc.)
• Remove sources of infection prior pruning (extir- • Minimize the number and size of new pruning
pation of symptomatic and dead vines) wounds
• Prune grapevines during dry weather • “Return cut” if needed should be done with the help of a
longer 2-year-old cane to prevent large wounds
• Remove pruning debris as soon as possible
(pruning debris mulching, compost, etc.) • Increase spur/cane length of pruned vines in order
to minimize fungal penetration into perennial wood
• Avoid depositing of pruning debris and/or dead vines in
areas approximate to the vineyard • Consecutive pruning of symptomatic and asymptomatic
vines is allowed due to irrelevant fungal inoculum trans-
Minimize new infections mission with pruning shears
• Pruning shears disinfection is a good hygiene practice,
• Preventative disease management, implemented but not of key importance to limit GTD spread
prior symptom development, is essential for a long-term • Implement double pruning, if not applicable replace with
productive vineyard early/late pruning
• Fungicides (biological and chemical) are efficient only as • Implement early/late pruning to minimize new
preventative treatments that limit new infections
• Coordination of work to minimize new infections - short Collective disease management

period of time between pruning and pruning
wound protection is crucial • Application of a single control method in GTD manage-
• Preventive protection of pruning wounds (physi- ment has only partial efficiency, implementation of
cal, biological, chemical) in a short interval after pruning more methods in disease management is essential.
• Adjustment of nozzles on pruning wound zone to achieve
better cover with fungicides (biological/chemical) Potential limitations
• Application of fungicides (biological and chemical) on • Technical knowledge
pruning wounds with high volumes of water
• Lack of equipment that contributes to high efficiency
• Clean properly tank of the sprayer prior application of (compost facility, trimmer for mechanical pruning, etc.)
Trichoderma spp. in order to avoid residual impact of
chemical fungicides on this microorganism (remember: • Availability of mastics and fungicides (biological and
Trichoderma are a group of fungi and chemical fungi- chemical) on the national market
cides have a negative impact on their activity!) • Cost-efficiency related to practice efficiency and pro-
duct value
Good pruning Wrong pruning
IPTPO (K. Diklić) IPTPO (K. Diklić)

Large wounds and
pruning cuts near pe-
rennial parts of grapevine
cordon and/or trunk induce
wood necrosis and poten-
tially lead to higher in-
fection rates with some
species of GTD fungi.
IPTPO (K. Diklić) IPTPO (K. Diklić)

Edwards, J., Laukart, N., Pascoe, I.G., (2001). In situ sporulation of
More information Phaeomoniella chlamydospora in the vineyard. Phytopathologia Mediter-
ranea, 40, 61–6.
KNOWLEDGE RESERVOIR: www.winetwork-data.eu
Elena, G., Luque, J. (2016). Pruning debris of grapevine as a potential
inoculum source of Diplodia seriata, causal agent of Botryosphaeria die-
VIDEO CLIPS: back. Eur. J. Plant Pathol., 144, 803-810.
1.Grapevine trunk diseases epidemiology and symptoms
(presentation by V. Mondello) Elena, G., Luque, J. (2016). Seasonal Susceptibility of Grapevine Pruning
Wounds and Cane Colonization in Catalonia, Spain Following Artificial
2.Pruning wound protection – experiences in Trichoderma
Infection with Diplodia seriata and Phaeomoniella chlamydospora. Plant
application Disease, 100(8), 1651-1659.
3.Double pruning
Geoffrion, R., Renaudin, I. (2002). Anti-esca pruning. A useful measure
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des Végétaux (France).
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Eutypa dieback and performance of ‘Cabernet Sauvignon’ grapevines. J.
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evaluation of their potential for spread of infection. Plant Dis., 99, 976- practices: the case of trunk diseases in California winegrape vineyards.
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touly, J.-P., Rey., P., Guérin-Dubrana, L. (2011). Impact of biotic and abio-
Bertsch, C., Ramírez-Suero, M., Magnin-Robert, M., Larignon, P., Chong, tic factors on the development of Esca decline disease. Integrated pro-
J., Abou-Mansour, E., Spagnolo, A., Clément, C., Fontaine, F. (2013). tection and production in viticulture, IOBC bulletin, 67(2011), 171-180.
Grapevine trunk diseases: complex and still poorly understood. Plant Pa-
thology, 62, 243-265. Lecomte, P., Louvet, G., Vacher, B., Guilbaud, P. (2006). Survival of fun-
gi associated with grapevine decline in pruned wood after composting.
Cahurel, J.-Y. (2009). Influence of training systems on wood diseases. IFV Phytopathol.Mediterr., 45, S127-S130.
Pôle Beaujolais, Bourgogne, Jura, Savoie.
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France. Plant Pathology, 47(4), 463-472.
Li, S., Bonneu, F., Chadoeuf, J., Picart, D., Gégout-Petit, A., Guérin-Du-
Cloete, M., Fourie, P.H., Ulrike, D.A.M.M., Crous, P.W., Mostert, L. (2011). brana, L. (2017). Spatial and temporal pattern analyses of esca grape-
Fungi associated with die-back symptoms of apple and pear trees, a vine disease in vineyards in France. Phytopathology, 107(1), 59-69.
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wood-streaking: two old and elusive diseases of grapevines. Plant di-
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L., Parrini, C. (2004). On the factors that may have influenced the esca
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(2012). Evaluation of fungicides for the management of Botryosphaeria
canker of grapevines. Plant Disease, 96(9), 1303-1308. Travadon, R., Lecomte, P., Diarra, B., Lawrence, D.P., Renault, D., Oje-
da, H., Rey, P., Baumgartner, K. (2016). Grapevine pruning systems and
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Pouzoulet, J., Pivovaroff, A.L., Santiago, L.S., Rolshausen, P. (2014). Can riaceae species isolated from grapevine cankers in California. Plant Di-
vessel dimension explain tolerance toward fungal vascular wilt diseases sease, 93(6), 584-592.
in woody plants? Lessons from Dutch elm disease and esca disease in
Van Niekerk, J.M., Halleen, F., Fourie, P.H. (2011). Temporal susceptibility
grapevine. Front. Plant Sci., 5, 253.
of grapevine pruning wounds to trunk pathogen infection in South African
grapevines. Phytopathol. Mediterr., 50(4), 139-150.
Ravaz, L. (1922). Le court-noué. Progres Agricole et Viticole, 76, 56.
Weber, E., Trouillas, F., Gubler, D. (2007). Double pruning of grapevines: a
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gninia minima perithecia in Esca-affected vineyards in California. Plant Enology and Viticulture. 58(1), 61-66.
Disease, 89, 867–71.
Work realized in common by the facili-

tators agents of Winetwork project. Data
came from practice through the help
of 219 interviews and from a review of
scientific litterature.
Thanks to Dr. Vincenzo Mondello and
Prof. Florence Fontaine who participated
to the development of this datasheet.
APPENDIX 2: GLOBAL VINEYARD STRATEGY TO PREVENT
GTDs INFECTION
www.winetwork.eu
13/27
TECHNICAL DATA SHEET1
GLOBAL VINEYARD STRATEGY TO PREVENT GTDs
Vineyard management practices to prevent GTDs

Technical data sheet
PLANTING PHASE
1. Planting a new vineyard
The first step for a healthy vineyard is to choose varieties that are less susceptible to develop trunk
diseases. Several studies demonstrated that the incidence of Esca is lower in Montepulciano and
Merlot and higher in Cabernet, Sangiovese, Cabernet sauvignon, Chardonnay, Sauvignon Blanc,
Riesling, Semillon, Trebbiano.
Figure 1: sensitivity degree of most common grape varieties
Rootstocks as well plays a role in susceptibility to GTDs: Vitis riparia 039-16 and Freedom have a
good degree of tolerance, like Vitis riparia X Vitis berlandieri. Similar degree of susceptibility is
reported between not grafted and grafted vines, on European or American rootstocks. Some authors
refer the positive influence of certain rootstocks, such as Rupestris, in the resistance to Esca, probably
due to the high tannin content in the plant that reduces the potential of infection. Since 2016 in Galicia
the susceptibility of autochthonous cultivars to Esca, Eytypa dieback and Botrysphaeria dieback is
under study. A 4-year study made in France assessed the different rootstocks impact on Esca foliar
symptoms expression: Riparia Gloire de Montpellier was the rootstock leading to less Esca foliar
symptoms. Other rootstocks, as the “101-14”, the “3309” and “Gravesac” tended to be more
sensitive, but effects could be reversed depending on meteorological conditions. Evaluation trials of
grapevine rootstocks against GTDs soilborne pathogens attested that the rootstock 110R was the most
susceptible to both black-foot and Petri diseases while the 161-49C rootstock resulted the most
tolerant to Petri disease infection.
Choice of the location, which will condition all subsequent vineyard grow and development, is also
relevant to minimize the damage caused by the GTDs. So, whenever possible, we must proceed the
grading that get slopes lower than 10%. If the slopes are higher, between 10% and 20%
earthmoving works would be important, so we must think about the possibility of planting along the
contour lines. With slopes higher than 20%, we already have to consider the need to make terraces.
Page 1
Overall the sites where vines grow easier, meaning exposition to South, preference to higher part of
the hills where winds keep moisture level low, have a positive impact on vine health status and
consequently reduce GTDs risk.
The planting period must be chosen carefully avoiding too late planting. The best period of time is
from late Autumn to early Spring, during the dormant plant season. In areas with cold winters, March
planting is preferable.
How to handle vines is also crucial: avoid leaving roots soaked in water over 24 hours before
planting and carefully water newly planted cuttings in order to avoid water stress (in both senses). At
planting pay attention not to break the roots and be careful that the root system is not be folded but
well stretched in all the available space in order to facilitate the best root development. In general is
important to avoid soil compaction. For this purpose don’t entry in the field with heavy machine when
the soil is soaked and prefer a double-layer cultivation.
It is shown an example of a tool designed by Ribeiro's own grower, consisting of a metal tube with a
channel for introducing the plant and thus being able to conduct it in depth without bending the roots
and in a straight way.
Figure 2: Planting with special homemade tool (courtesy of Ángel González of Beade winery in Ribeiro D.O. Galicia, Spain)
At planting time the inoculation with Trichoderma-based products (T. harzianum, T. atroviride, T.
asperellum, T. gamsii) could be recommended. Before planting, vines roots are soaked for an hour in a
solution containing Trichoderma. Trichoderma improves root growth and stress resistance in colonized
plants, which would possibly make plants less sensitive to wood diseases. Vines can also be soaked for
50 minutes in a water solution containing cyprodinil and fludioxonil or metiram and pyraclostrobin.
These mixtures reduces incidence and severity of black foot disease and Botryosphaeria dieback.
The best period of the year for field grafting is spring, or at the flowering time for the Northern
regions, when the vines are starting to push new growth. Try to graft when the vines are dry (days with
no rain) to reduce the risk of disease’s infection.
The choice of the training system should avoid over simplification and favor a physiologically sound
pruning, allowing a smooth sap circulation. It is to prefer less severe pruning, with small pruning
Page 2
wounds and a reduced interference with vessels development. Long pruning systems should be
preferred and several evidences support that “alberello” and Guyot systems should be preferred.
When planting a new vineyard it is extremely important to promote a right vertical growing of the
trunk, binding the new plant to a good stake: a vertical trunk is less sensitive to machinery damage
used for weed-control under-row.
Pruning in the formation phase

It is important to adopt pruning preventive measures since the beginning of the vineyard life. In
general, all practices that cause stress in plants should be avoided. A reasonable pace to establish the
definitive structure of the plant, avoiding big pruning wounds, and respecting sap flow paths is
essential.
High density systems increase GTDs risks, so the balance among production, quality and health needs
a compromise, considering the forecasted life span of the vineyard.
2. Soil preparation
The first areas of the vineyard where symptoms of GTDs appear are identified as particularly dry or
areas where water-logging persists for long periods, combined with poor nutritional conditions. Before
planting it is advisable to make soil analyses to know the mineral and organic substance content and
then compare the data with the mean values of the territory. Improvement of general soil conditions,
starting from the physical structure down to nutrients availability it is recommended if poor nutritional
conditions exist. Moreover, it must be verified that the soil is not infected by analysis in an authorized
laboratory. The fundamental problems in soil come mainly from the fungi that cause root rot such as
Armillaria mellea and Rosellinia necatrix and from the Nematodes.
Before planting it is important to remove all the previous pruning debris, especially if GTDs were
detected. In the case of previously existing plantations, it is advisable to wait before replanting at
least 3-4 year, (better 6 or 7). A useful practice, supporting the residues degradation and the related
pathogens is the green manuring, as it adds organic matter that facilitates microbial activity.
Furthermore, the green manure roots development helps the soil structuring. Specific crops may be used,
such as rape (Brassica napus L.) or mustard (Brassica juncea L.), that produce allelopatic substances
(volatile isothiocyanates) able to suppress/reduce pathogenic fungi. As an alternative, mustard meal
can be incorporated into the soil, leading to comparable results.
Planting is also the phase in which future management possibilities are set and their potential to reduce
wood diseases out-break should be taken into account.
Page 3
Figure 3: Use of brassica rapa in the vineyard (IFV South-West)
These practices are:
- soil erosion reduction through appropriate orientation of the vineyard rows and soil protection by
cover crops
- an effective drainage system
- improvement of soil structure by increasing or maintaining a good level organic matter through
green manuring, addition of compost and other organic materials
- increase of phosphorus and potassium availability, linked to the previous point and to soil
structure.
If the previous use was shrub must be carried out a deep ploughing or subsoiling preferably that will
fragment the horizons of the soil in a vertical way facilitating water’s drainage and allowing the
proliferation of roots of depth vines.
3. How to check and to handle nursery materials
The use of certified planting materials is always recommended. A healthy vine has a higher potential
to react to infections, granting an easier start of the crop and a more sustainable management of the
vineyard. When receiving the vines from nursery, a visual check for necrosis in the wood is advisable
and, if found, a microbiological analysis is recommended. However, when rooted vines show very large
necrosis, even if they are not colonized by pathogenic fungi, they have unpredictable performances
over time, for instance irregular sprouting and/or a stunted vigor. In young vineyards wood decay
frequently starts from the grafting point, especially if grafted in the splitting of green shoots and the
graft is just above the ground-level. The omega grafting is the most extensively used but it leads to a
30-50% of dead wood. The best choice for GTDs prevention is the bud grafting. Be aware that
grafting cause wounds that increase the possibility for the pathogenic fungi to enter into the plant.
Wounds should be protected with sprayed or painted formulations.
Page 4
GROWING PHASE
1. Pruning
Pruning time is important since it could influence the vine sensitivity to pathogens and abiotic
disorders. Pruning should be trimmed according to vine physiology and be implemented during dry
and non-windy periods. Regarding the best pruning moment there is no agreement. Late pruning in
the dormant season (as close as possible to bud-break) was a recommended cultural practice, since the
wounds heal faster with high degree-day temperatures. Nevertheless, recent studies revealed that the
rate of natural infection of pruning wounds was lower in early pruning (Autumn) than in late pruning
(Winter). Nevertheless, experiences report different outcomes on the topic and in some areas early
pruning is preferred. The susceptibility of the wound is mostly influenced by relative humidity and
rainfall. Double pruning or pre-pruning is enhanced by growers to speed up final pruning and to
reduce disease incidence in spur-pruned vineyards, also considering that spores need only up to 5
hours of wetness (rain) to infect wounds. Sanitation methods are often complemented with pruning
wounds protection from frost or biotic attacks through the application of fungicides, biological
formulations or both in alternation.
Figure 5. The best pruning technique assure a continuity of the flow and respect of the vascular system. With kind
permission of «I preparatory d’uva» www.simonitesirch.it
Some studies demonstrated that the infections after pruning can be significantly reduced by using a
single paste application with a mixture of benomyl, pyraclostrobin, tebuconazole or thiophanate-
methyl. Anyway, in order to be effective, the products must be applied directly onto the wounds. Some
other studies demonstrate that paste application were no effective for Esca or Botrysphaeria species
but only for Eutypa.
Cuts close to the perennial wood (clear-cuts) (usually produced by electric pruning scissors) should be
avoided in order to reduce the formation of wood drying up cones. Pruning systems that allow a
better sap flow circulation over the years (e.g. Guyot-Poussard) could be considered should be
preferred.
Page 5
Mechanization vs hand-made operations
The excessive simplification of training system (mechanical pruning, harvest, etc.) is probably, at
present, one of the most harmful reasons that involve GTDs, as each operation risks to open wounds
and give way to infections.
For the sake of plant health, hand-made operations should be preferred. Nevertheless, economic and
organizational aspects should be taken into account, leading to prefer a shorter vineyard life-span
than a longer and healthier one.
2. Weed control in the under-row: respect the trunk
The increasing use of equipment for under-row tillage, replacing herbicides, may cause injury to the
trunk if it is not carried out with the right attention, facilitating GTD’s infections. In order to reduce this
risk, the right equipment should be chosen and carefully tuned to the specific vineyard. The feeler
sensitivity commanding the movement back from the row must be quite high: it’s better to have some
grass left back than an injured trunk. In the case of working absence of a feeler system (circular or
multi-petal working heads) profiles that could cause injury should be avoided. For the same reasons,
de-suckering machines should not peel the vines bark (decorticate).
Topping, canopy thinning, leaf removal machines (green canopy management) do not seem to impact
as operating on still green shoots, while the risk may arise in case of too late interventions on woody
branches. Anyway, there is currently no evidence about it.
3. Soil management
Plant stress caused by unbalanced nutrition, poor drainage, soil compaction, heavy crop loads on
young plants, planting of vines in poorly prepared soil and improper plant holes play an important
part in the development of GTDs especially on the foliar symptoms expression. Since the soil is the
main source of inoculum for soilborne fungi, disease management practices based on soil correct
management is key in the prevention of GTDs diseases.
In general a well-structured soil, where air and water easily circulate and is never water logged or
saturated is a key aspect in prevention.
In the first five years it is essential to prevent an excessive plant vigor, allowing the plant to exploit
most of its resources for the development of a good and deep root and vascular system. As a
consequence, application of high doses of nitrogen should be avoided. A high C/N ratio of soil
organic matter reduces plant vigor and favors secondary metabolism, increasing the production,
among others, of polyphenols that increase plant natural resilience towards pathogens. A reduced
vigor (and the consequent delayed production), allowing a more balanced vine growth, could help the
plant to be less susceptible to GTDs.
In general the use of cover crops (not only leguminous species) and, even better, permanent coverage
of the area between the rows with herbaceous species balances nutrients availability and improves
soil structure. In the choice of the mixture composing the coverage it is important to include deep
rooting species (i.e. lucerne) as it improves lower layers structure and air circulation. Moreover, lucerne
Page 6
increases phosphorus availability and it avoids soil compaction due to mechanical means circulation. In
case of lack of nitrogen, green manures rich in leguminous species can be preferred for a period of
time, since balance is set.
In any case, a good availability of organic matter should always be a goal. Besides green manures,
compost and organic fertilizers should be added to balance yearly mineralization. Compost can be
produced with manure, mowed grass, wine-making resides and pruning residues. In the latter case,
care should be payed in avoiding infected materials or to run a long fermentation process able to
inactivate pathogens.
Soil tillage must avoid soil compaction. Over-watering and water-logging should be avoided as
well as water stress, which in particular conditions can lead to a wood efficiency and functionality
impairment and to an increased susceptibility to fungal infections. Moreover, ground fractures just
below the surface due to drought periods, can cause root breaking and desiccation but also fresh
wounds that are the main infection routes.
4. Water management and irrigation

High humidity level in the soil (and in the air), together with warm climate, provides the optimal
condition for fungi propagation and development, especially for soil-borne pathogens.
On the other side, severe water stress could cause a higher expression level of GTD symptoms.
Drip irrigation must be regulated to avoid both critical conditions. The daytime is more advisable
than night-time in Summer irrigation. The so called “mild stress”, applied to promote a good ripening is
reasonable, but too extreme in“thirst” of the vineyard may lead to the GTDs symptoms appearance in
affected vineyards.
In case of excess vigor and/or an excess yield, due to high nitrogen fertilization, use of vigorous
rootstocks and continuous soil tilling is always dangerous for GTD risk but mostly in case of dry-land
farming, because in a dry season this excess increases the water stress establishment and the explosion
of GTD symptoms.
5. Biological control to prevent infections

Trichoderma exhibits antagonistic activity towards other microorganisms, especially soil-borne ones.
Healthy vines can be inoculated with these fungi to colonize the woody tissues of the cordon and the
trunk up to few centimeter under the treated wounds. Trichoderma is applicate on the vegetation and
the fungus colonize the plant competing for the same ecological niche with the other fungal pathogens
stimulating a better defense response towards GTDs pathogens. The use of various species of the
fungal genera Trichoderma (T. harzianum, T. gamsii, T. atroviride, T. asperellum) to protect pruning
wounds came into use with various commercial preparations. The fungus colonizes the pruning wounds
forming a barrier to the pathogens penetration. Its action is only preventive and it is linked to a
various number of variables affecting the biological habit of the fungus and its capacity for
colonization. Complete colonization require a certain time, during which the grapevine is sensible to
infections and Trichoderma may be washed off in case of rainfall. The ideal is to spray Trichoderma as
soon as possible after pruning.
Recommendation: prune by plots or sectors and as soon as one plot is completed, spray
Trichoderma.
Page 7
PRODUCTION PHASE
1. Annual pruning
Same concepts reported for the growing phase worth true in the production phase.
2. Debris management
In order to reduce diseases spreading, infected wood should be removed from the field, either
burned or composted. It is particularly important in the case of old wood (branches, trunks), while one-
year wood is less dangerous.
Chopping and burying the pruning wood could create a dangerous inoculum in the soil.
Some practices to reduce the inoculum are suggested:
• Remove dead vines, diseased vines or dead parts of alive vines (dead shoots).
• Remove or burn them or protect them away from rain. GTDs inocula are found at surface of
the wood in all part of grapevine (arms, trunk).
• Remove debris before pruning, better if together with observe diseased vines at stage of
8-12 leaves for Eutypa dieback and before harvest for Esca and Botryosphaeria dieback
• Consider that some pathogens as Botryosphaeriaceae could resist in pruning debris for
more than 3 years.
Another suggested practice is the composting, beneficial to soil organic matter increase. Its
management should consider the following:
• peak temperature of 64-70°C and 21 days of fermentation in general ensure the

reduction of pathogens below detection limits, but this practice is not tested on GTDs
pathogens
• compost of 3 years old made of pruning debris, sheep manure, leaves and grass residues
(temperatures around 50-60°C) allow to eradicate GTDs pathogens (Eutypa lata,
Phaeomoniella chlamydospora sp, Phaeoacremonium. minimum, Botryosphaeria sp.) limiting
mycelial development. Composted material can be reintroduced in vineyard with no risk of
contamination.
3. Fertilization
During the production phase a balanced nutrition to assure a balanced growth and a limited vigor is
the goal. Thus, incidence of wood diseases can be reduced by a moderate fertilization that grants the
plant the resources for production but also for self-defense. Indeed, an excessive vegetative growth
affects both the plant lignification and the ability of the plant to self- protect. Moreover, an increased
plant vigor needs more severe pruning that causes large wounds facilitates infections. A
moderate nitrogen availability and limited irrigation are advisable.
It is demonstrated that foliar applications of nutrients influence the development of GTDs foliar
symptoms. For example, foliar application of a mixture of calcium chloride, magnesium nitrate and
Fucales seaweed extract during three years led to a significant reduction of symptoms development in
Page 8
the vine treated with the full mixture. Both quantity and quality of grapes from the treated vines
increased, while no phytotoxic or other unwanted effects on grape growth were detected.
4. The use of Trichoderma and other biocontrol agents

For the prevention of GTDs, Trichoderma harzianum and T. atroviride treatments during the whole plant
life are recommended. Another possibility is the induction of grapevine self-defence systems using
other biocontrol agents. A scientific study ascertained that necrosis, produced by Pa. chlamydospora
(one of the Esca disease pathogens), were reduced up to 50% when Pythium oligandrum colonized the
root system of grapevine cuttings. A commercial product containing this biocontrol agent is currently
available.
Other products, based on a mix of arbuscular-mycorrhizal fungi, if inoculated in vines could reduce the
susceptibility to GTDs.
5. Soil management and Weed management in the under-row

The same concepts reported for the growing phase worth true in the production phase.
6. Water management and irrigation

It has been evidenced that a water-stressed vineyard in a warm, dry environment may be more
susceptible to infections through pruning wounds by Eutypa lata than vines receiving regular
irrigation. Internal wood symptoms are not related to foliar symptoms severity or to the temperature
and moisture combination. Water-stressed vines have significantly lower photosynthetic rates and
lower levels of stomatal conductance compared to those receiving optimal irrigation, indicating that
these plants experienced significantly higher levels of physiological stress. Fungal diseases which enter
the plants from the pruning wounds produced some external symptoms and the lesion length are
significantly longer in pruning wounds of stems from plants in the lowest irrigation regime, with lesion
length declining linearly with the increasing irrigation volume. These results clearly indicate that when a
grapevine is exposed to water stress, colonization and disease expression by Botryosphaeriaceae
spp. are much more severe.
Practical recommendations reported for the growing phase worth true also in the production phase.
7. Trunk renewal practices

The decision to replace vines is not only based on economic factors, since there are agronomic ones
(establishment of new vines, yield, uniformity quality of grape, etc.) which could affect the viability of
Page 9
the new plants. Some practices, such as trunk renewal, re-grafting and trunk cleaning (surgery) could
be also considered. An inspection to identify symptomatic vines in early stages is recommended.
The inspection time depends on the predominant trunk diseases in the vineyard. Foliar symptoms of
Eutypa dieback and Botryosphaeria dieback are visible in Spring while the Esca ones develop starting
approximately in mid-June. Dead spurs and stunted shoots are best observed later in the growing
season, when vegetative growth stops.
Symptomatic vines must be marked in order to evaluate the degree of infection in a plot, to prune
infected plants separately from others or to follow up and evaluate implemented actions.
The trunk renewal practice consists in the recovering of a diseased vine by replacing the infected trunk
with a new one, utilizing a sucker at its base. Scientific studies have shown successful results when
applied against Eutypa dieback and other trunk diseases.
The experience in many countries shows that the sooner the trunk renewal begins, the more
successful it will be in controlling the disease spread and yield loss. This practice allows recovering
the plant and taking advantage of the root system of the damaged vine, thus mitigating the losses
by damages of wood disease and maintaining the productivity of vineyard. Two suckers can be
used to form two trunks, a helpful insurance against new infections, or an extra possibility in case of
damage.
Figure 6: Suckers kept to renew the trunk (Photo courtesy of Lucía &
Manolo Vilerma.)
Page 10
Figure 7. A vine after the trunk renewal practice. (Photo courtesy of Lucía & Manolo
Vilerma.)
Trunk cleaning removes from trunk or arms the rotten wood that disturbs the circulation of sap. It
means to open the trunk or arms, to remove the affected wood, keeping only the external part of the
wood or cambium. The cut is always made above the graft point and about 20 cm below the any
wood staining. Early implementation, as soon as first symptoms appear, is recommended, if done in
June it allows to harvest in the current year.
Figure 8: Appereance of the trunk after trunk cleaning (IFV Alsace)
Regrafting on rootstock and overgrafting on existing cultivars are other procedures that can be used
to recover vines affected by GTDs, as soon as it can be assured that vine’s material to be recovered is
healthy.
Regrafting can be taken either from a sucker which has sprouted from a rootstock or from the
rootstock directly. In the first situation, it is usually applied when sap is flowing, while on vegetative
growth, before phenological stage BBCH 75 with a pea’s sized grape, either by a bark grafting or by
Page 11
a whip grafting. In the second situation it is taken at the end of the dormant period and before bud
break, normally by a cleft grafting.
The other method of overgrafting is made using the same techniques and periods described above on
regrafting, but in this case it is always applied on the cultivar’s trunk.
Both situations take advantage of vine´s good rooting system, which implies lower harvest loss and
faster reaching of full production. Through grafting, vines can reach adult’s production level in three
year’s time and with the same grape’s quality. However, it must also be taken into account that these
techniques take an important amount of time and work and there can also be a number of vines in
which grafts are not welded.
S O U R C E O F I N F O R M AT I O N S :
• Almeida F., 2007. Technical notes 2 “Grapevine wood diseases. Eutypa dieback and Esca”. ADVID
Technical notes, 14 pp.
• Biribent M., 2015. L’innesto in campo e la longevità dei vigneti. Progetto SALVE: Ruolo del
materiale di propagazione per la salvaguardia del patrimonio viticolo campano. Comune di Lapio
(AV), May 6th, 2015.
• Bongiovanni S, Marzocchi L., 2013. Prevenzione integrata del mal dell’esca. Terre&Vita, 15, 46-
50.
• Bottura M., Aldrighetti C., 2003. Mal dell’esca della vite: malattia da non sottovalutare. Terra
Trentina, 4, 35-37.
• Calzarano F., Di Marco S., 2007. Wood discoloration and decay in grapevines with esca proper
and their relationship with foliar symptoms. Phytopatologia mediterranea, 46, 96-101.
• Calzarano F., Di Marco S., D’Agostino V., Schiff S., Mugnai L., 2014. Grapevine leaf stripe disease
symptoms (esca complex) are reduced by a nutrients and seaweed mixture. Phytopathologia
Mediterranea (2014) 53,3, 543-558.
• Corti G., Agnelli A., Cuniglio R., Ricci F., Panichi M., 2004. Suolo e mal dell’esca della vite: il punto
di inizio delle indagini. L’Informatore Agrario, 12, 79-84.
• Curti G, Cuniglio R., 1999. Vite: caratteristiche del suolo e incidenza del mal dell’esca.
L’Informatore agrario, 40, 64- 67.
• Di Marco S., 2009. Esca e materiale di propagazione della vite: aggiornamento sulle recenti
acquisizioni scientifiche. Convegno Vitis, Rauscedo, November 20th, 2009.
• Fontaine F., Gramaje D., Armengol J., Smart R., Nagy Z.A., Borgo M., Rego C., Corio-Costet M.-F.,
2016. Grapevine Trunk diseases. A review. OIV, 24 pp.
• Gramaje D., García-Jimenéz J., Armengol J., 2010. Grapevine rootstock susceptibility to fungi
associated with Petri disease and esca under field conditions. Am. J. Enol. Vitic., 61, 512-520.
• Gramaje D., Di Marco S., 2015. Identify practices likely to have impacts on grapevine trunk
diseases infections: a European nursey survey. Phyopatologia mediterranea, 54 (2), 313-324.
• Gramaje D., Alaniz S., Abad-Campos P., García-Jiménez J., Armengol J. 2016. Evaluation of
grapevine rootstocks against soilborne pathogens associated with trunk diseases. Acta
Horticulturae, 1136: 245-249.
• Groupe national maladies du bois, 2007. Note nationale maladies du bois, 5pp.
• Gubler W.D., Baumgartner K., Browne G.T., Eskalen A., Rooney-Latham S., Petit E., Bayramian L.A.,
2004. Root diseases of grapevines in California and their control. Australasian Plant Pathology, 33,
157-165.
• Larignon P., 2004. La constitution d’un groupe international de travail sur les maladies du bois et
les premiers résultats des experimentations menées par l’ITV en laboratoire et en pépinières. Les
maladies du bois en Midi-Pyrénées, 24-27.
• Lecomte P., Louvet G., Vacher B., Guilbaud P., 2006. Survival of fungi associated with grapevine
decline in pruned wood after composting. Phtyopathologia Mediterranea 45, S127-S130.
Page 12
• Lecomte P;, Diarra B., Chevrier C., 2015. Role de la conduit sur le dévveloppement des maladies
du bois de la vigne. Compte rendu des journées nationales maladies du nois, 17 et 18 november
2015, Université de Haute Alsace, 57p, 20-21.
• Niekerk J.M., Strever A.E., du Toit P.G., Halleen F., 2011. Influence of water stress on
Botryosphaeriacea disease expression in grapevines. Phytopathologia Mediterranea (2011) 50,
S151-S165
• Noble R. and Robets S.J., 2004. Eradication of plant pathogens and nematodes during
composting : a review. Plant Pathology 53, 548-568
• Mugnai L., 1999. Il mal dell’esca della vite. L’Informatore agrario, 15, 77-81.
• Mugnai L., 2016. Available tools and approaches for GTD control in the vineyard. Convegno
Winetwork “Scienza & Pratica vs Malattie del legno & Flavescenza Dorata, November 9th, 2016,
Conegliano (TV).
• Quaglia M., Covarelli L., Zazzerini A., 2009. Epidemiological survey on esca disease in Umbria,
central Italy. Phytopatologia mediterranea, 48, 84-91.
• Rego C., Nascimento T., Cabral A., Silva M.-J., Oliveira H., 2009. Control of grapevine wood fungi
in commercial nurseries. Phytopatologia Meditteranea, 48, 128-135.
• Roby J.P., Mary S., Lecomte P., Laveau C., 2015. Rootstock impact on foliar symptom expression of
esca on Vitis vinifera cv. Cabernet sauvignon. 5pp.
• Serra S., Peretto R., 2015. Le malattie del legno di origine fungina. Agenzia Laore e Università di
Sassari, 40 pp.
• Sosnowski M.R., Luque J., Loschiavo A.P., Martos S., Garcia-Figueres F., Wicks T., Scott E.S., 2011.
Studies on the effect of water and temperature stress on grapevine inoculated with Eutypa lata.
Phytopathologia Mediterranea (2011) 50, S127-138.
• Sportelli G.F., 2008. Il mal dell’esca aggredisce anche le viti giovani. Terra&Vita, 14, 54-58.
• Viret O., 2014. Malattie del legno, l’unica soluzione è prevenirle. L’Informatore Agrario, Suppl. 13,
11-13.
• Whitelaw-Weckert M., Rahman L., Cappello J., Bartrop K., 2014. Preliminary findings on the
grapevine yield response to Brassica biofumigation soil treatments. Phytopatologia mediterranea,
53(3), 587.
• Yacoub A., Gerbore J., Magnin N., Vallance J., Grizard D., Guyoneaud R., P. Rey P., 2014. Induction
of grapevine defence systems using the oomycete Phytium oligandrum against a pathogenic fungus
involved in Esca. Pahytopatologia Mediterranea, 53(3), 574-575.
• http://www.maladie-du-bois-vigne.fr/Maladies-du-bois/L-esca/Protection-au-vignoble
Page 13
APPENDIX 3: TRICHODERMA APPLICATION FOR PROTECTING
GRAPEVINE PRUNING WOUNDS
www.winetwork.eu
14/27
Trichoderma application
for protecting grapevine pruning wounds

Introduction Practical application
Trunk diseases significantly limit the productivity and longe- The different strains of Trichoderma spp. are able to colonise
vity of vineyards in most of the winegrowing regions all over about 1-2 cm of the pruning wounds and prevent
the world. Trunk diseases attack the permanent woody the penetration (into the wood) of pathogens associated
structure of the vine, including the trunk, cordons and spurs with GTDs. The colonisation of grapevine pruning wounds by
(Baumgartner, 2013). The pathogens - a set of taxonomi- the Trichoderma spp. depends on the physiological state
cally-unrelated Ascomycete fungi - associated with of the vines as well as the weather conditions at pruning.
grapevine trunk diseases are able to infect healthy vines The pruning season coincides with the period of pathogen
mainly by pruning wounds and these wounds can remain spore release which usually originates from infected wood.
susceptible for several months. It is important to highlight Wounds may remain susceptible for a long time (up
that there are no curative methods to control GTDs; the only to 4 months or more, according to the GTD), but the most
way is to prevent or limit the infection of wood di- critical time for infection ranges from 2 to 8 weeks af-
seases using different cultural practices. The pre- ter pruning (Eskalen et al. 2007, Van Niekerk et al. 2011b).
vention of wound infection applying biocontrol agents is
one of an alternative technique to control trunk diseases.
Species of the genus Trichoderma (an ascomycete fun- 1- Time of application
gus, originally present in the soil) have been investigated
several times as a potential biocontrol agent by spatial and
nutritive competition. Normally, Trichoderma spp. are not limited by climatic condi-
tions, being able to start wound colonization at 10° C, but the
time of the treatment could improve its efficiency in coloni-
Application area zing wounds and thus, its protection capability. The correct
timing is above 0°C temperature, though some Trichoderma
species require higher temperature (exceeds 10 °C). It is
Use of trichoderma to protect pruning wounds is very popular important to highlight that Trichoderma spp. as a biocontrol
in Europe and implement on the field by many winegrowers. agent is susceptible to the frost. The best timing could be as
soon as possible after pruning, to limit the wound sus-
ceptibility period to new GTDs infections. Different studies
attested that better colonization results could be achieved
with treatments done within 5 or 6 hours after pruning (Har-
vey et al., 2006, Mutawila et al., 2016).
Some producers recommend distributing Trichoderma pro-

ducts during bleeding, since the sap presence helps the an-
tagonist in colonizing the wounds faster. At the same time it
is important to check the weather forecast before the appli-
cation because heavy rain can interfere with the beginning
of colonization, washing away the spores.
Scientists recommend to plant vines that have been inocu-

lated with Trichoderma spp in nursery during the propagation
process and repeat field treatment 2 or 3 years after planting.
Figure 1: European wine-growing areas where Trichoderma Then it is highly recommended to repeat the application
application is applied, red dot (result from Winetwork inter-
views). White dots are showing project’s partners .
each year thereafter (Sosnowski, 2016). Both small and
large wounds should be treated with the biocontrol agent
either by spraying or painting, according to the economic Outcomes
possibilities or the value of the vineyard.
Among Trichoderma species and strains, several are used
2- Mode of application in European countries in pruning wound protection: Tri-
choderma atroviride SC1 and I1237, Trichoderma asperel-
Preventive wound protection practices should start lum ICC012, Trichoderma gamsii ICC 080 and Trichoderma
in 1-year-old grapevines following the first pruning and harzianum ICC012 :
continue each year thereafter (Sosnowski, 2016). Both
the small and the large wounds should be treated with the • Trichoderma atroviride SC1 has been isolated from dead
biocontrol agent, using a canopy sprayer with nozzles targe- hazelnut wood and selected for its high colonization capabi-
ting the cordon (Sosnowski, 2016). When canopy sprayers lity and its high productivity of Lytic enzimes
are used, maximum coverage of wounds can be achie- (chitinases, proteases and cellulases). Trichoderma atrovi-
ved by turning off fans (no air), applying high water rates ride SC1 is highly competitive and efficiency antagonizes
at low pressure, selecting spray nozzles that produce large Phaeoacremonium minimum and Phaeomoniella chlamy-
droplet size and focussing nozzles towards the pruning dospora so is able to reduce the yearly infections on the pa-
wound zone. thogens associated to esca disease (D’Enjoy et al., 2016.)
• Trichoderma atroviride I1237 has the ability to fast colo-
Different canopy sprayers (modified weed sprayer, recy-
nize pruning wounds, to compete with pathogenic fungi for
cle sprayer, tangential sprayer and air-shear sprayer) have
nutrients and space and properties of antibiosis and myco-
been tested in cordon- and in cane pruned vines varying the
parasitism.
amount of water volume. According to the obtained results,
it is important to select adequate sprayer and the required • Trichoderma asperellum and Trichoderma gamsii ICC 080
amount of water to achieve the maximum coverage of the can have an effect on GTDs pathogens (especially on Phaeo-
vines. moniella chlamydospora) at 10°C and 15°C respectively.
When preparing for the treatment it is highly recommended Both species remain viable at 5°C.
to clean carefully the tank from previous fungicide resi-
dues in order to not ‘disactivate’ Trichoderma. For future practical applications, experimental trials should
be carried out to confirm its efficacy with a wide com-
One of the most important obstacles for the use and diffu- bination of application conditions.
sion of Trichoderma is often related to the variable results
observed by winegrowers. Indeed, numerous factors could The effectiveness of protection based on Trichoderma spp.
influence the biocontrol capability of a Trichoderma-based treatments depends on the ability of these fungi to colonize
product, namely the Trichoderma species utilized, the me- grapevine pruning wounds (John et al., 2008). For a com-
thod used for its distribution, the phenological stage of the plete colonization of the wound, trichoderma species usually
vines, the time between pruning and the Trichoderma treat- need some time, and during which grapevine is susceptible
ment, the interaction of the antagonist with host plant and, to infection from GTDs pathogens and to washing off by rain-
least but not last, with environmental factors (Di Marco et fall. However, more field tests are needed and necessary to
al., 2004.). Furthermore, the biocontrol activity could vary conclude on their effect on the short and long term and to
according to the different cultivar, (Mutawila et al. 2011a). All determine how it could be optimied by a combination of other
these factors, if not properly managed or not take in consi- management strategies (such as combination with other bio-
deration, could lead to unsatisfactory results. logical or chemical products, remedial surgery, reducing the
Thus it is important to not consider a Trichoderma treatment number and size of pruning wounds and application of sani-
similar to a chemical one. tation methods) (Bertsch et al., 2013).
Treatment can also be applied with backpack sprayer and
the spray need to be directed on the wounds surface and
well cover the wounds (Fig.3) on the entire grapevine.
Trichoderma atroviride SC1 (DLR Rheinpfalz)

Some scientific elements
One way to control grapevine trunk diseases is to protect

pruning wounds with fungicide applications, which can be
problematic because of the limited number of registered
products (not authorized in all european countries), the dif-
ficulty for these products to control numerous taxonomical-
ly unrelated organisms, the challenge of these products to
protect during the entire period of wound susceptibility and
the difficulties and costs associated with hand application of
protection treatments (Rolshausen et al., 2010).
The integration of fungicide and biological wound protection
could provide better control, but is limited by the susceptibi-
lity of the biocontrol agents to the fungicides.
The major way of managing trunk diseases in field grape-
vines is to prevent pathogen entry through pruning wounds.
Product for wound protection should be effective against the
whole range of trunk pathogens while also protecting the
wound for the whole period of wound susceptibility. Gene-
rally, the goal for pruning wound treatments is to inhibit my-
celial growth on the wound itself and/or physically seal the
wood to prevent infection (Newsome, 2012.). Trichoderma
are well known as fungi that exhibit antagonistic activity and
hyper-parasitism in regard to other microorganisms and it is
used for biological control against several diseases. Although
their mode of action is not fully understood, they seem to
be associated with mycoparasitism, the production of inhi-
bitory compounds, competition for nutrients and space with
pathogenic fungi, stimulation of plant growth and enhanced
host resistance (Di Marco et al., 2004). Since 2000s, several
trials were conducted in order to evaluate the efficacy of Tri-
choderma spp to control GTDs pathogens (table 1). Results
of these studies globally showed that Trichoderma spp. have
a partial efficiency according to assessment methods used
in controlling the main GTDs pathogens on both pruning
wounds in the field and cuttings at nursery, avoiding new in-
fections. Furthermore, thanks to its broad spectrum activity,
Trichoderma is able to delay infections of a wide range of
GTDs pathogens, staying viable in the woody tissues below
wound up to 1 year. Being a “living” product, its efficiency
Figure 3: Treatment of pruning wounds in a vineyard damaged by could be influenced by the environment. In particular wound
Esca (Eszterházy Károly University, N. Burghardt)
colonization capability and persistence of the Trichoderma
species may depend on intrinsic wound factors and hence
may vary between cultivars and on the vine physiological
stage in which Trichoderma is applied (Bruez et al, 2014; Di
Botryosphaeria dieback Eutypa dieback Esca complex
T. harzianum, T. atroviride, and Trichoderma spp Trichoderma spp
Benzimidazole-resistant mutant T. harzianum, T. atroviride, Benzimi- T. harzianum, T. atroviride, T. longibra-
strain dazole-resistant mutant strain chiatum
TESTED: pruning wound pro- TESTED: for Eutypa toxic metabo- and Benzimidazole-resistant mutant
tection lites degradation activity strain
for pruning wound protection TESTED: pruning wound protection
Bacillus subtilis EE isolate Bacillus subtilis EE isolate Bacillus subtilis EE isolate

TESTED: pruning wound protection TESTED: pruning wound protec- TESTED: pruning wound protection
tion
Pythium oligandrum
TESTED: induced resistance by root
colonization
Table 1: BCAs used to control GTDs (Esca, Botryosphaeria and Eutypa dieback)
Marco, 2007). Furthermore, the Trichoderma wound protec-

tion effect also depends on its interaction with the grapevine,
since it is not only due to the direct suppressive effect of
Trichoderma on pathogens, as reported by some researchers
(Mutawila et al, 2011).
A recent study (Aloi et al., 2014) showed the ability of Tri-
choderma gamsii+Trichoderma asperellum in reducing the
incidence of Esca symptoms when applied as wound pro-
tectant.
Key points for success

Trichoderma have a preventive effect on the infection of grape-
vine trunk diseases pathogens, to maximise its preventive effect,
several conditions need to be respected:
• Application of Trichoderma-based product as soon as Trichoderma species (DLR Rheinpfalz)

possible after pruning
• Application can be done either by sprayer (canopy sprayer or
backpack sprayer) and paintbrush.
• Respect Trichoderma strain characteristics (tempe- More information on
rature during application) and if possible apply the product
on dry conditions and before rain. www.winetwork-data.eu
• to maximise preventive action, start the application of Tric-
hderma on the first year and renew each winter at the
Technical datasheets: Good pruning practices
pruning period
Global vineyard strategy to manage GTDs
Video seminars:
• Scientific overview of grapevine trunk diseases (Dr. Vincenzo Mon-
dello, URCA)
• Symptomatology and epidemiology of grapevine trunk diseases
(Dr. Vincenzo Mondello, URCA)
Source of information
Aloi C., G. Bigot G., P.P. Bortolotti P.P., M. Cotromino M., S. Di Marco S., Eutypa dieback) Winetwork project SWG meeting minutes (2016)
F. Faccini F., A. Montermini A., L. Mugnai L., R. Nannini R., F. Osti F., F.
Mugnai L. What preventative measures could growers take to prevent
Reggiori F., 2014. Remedier® (Trichoderma Asperellum e Trichoder-
the entry of GTD agents into a vineyard? –Presentation at Wineskills
ma Gamsii): nuova opportunità di contenimento del complesso del mal
Masterclass on Grapevine Trunk Disease (2012)
dell’Esca della vite. Risultati di quattro anni di sperimentazione in Italia.
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methods of grapevine pruning wound protection BioControl (2015)
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vine wood from treated pruning wounds, New Zealand Plant Protec-
grapevine trunk diseases NZ wine project outcome (2016)
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John S., Wicks TJ, Hunt JS, Scott ES, Colonisation of grapevine wood
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Mondello V. BCAs used to control GTDs (Esca, Botryosphaeria and entomology.cornell.edu/pathogens/trichoderma.php
Work realized in common by the faci-

litators agents of Winetwork project.
Data came from practice through the
help of 219 interviews and from a
review of scientific litterature.
APPENDIX 4: A GUIDE OF GOOD PRACTICES FOR REGIONS
WITHOUT FLAVESCENCE DORÉE
www.winetwork.eu
15/27
1
Vineyard infected with ‘Flavescence dorée’ phytoplasma
A GUIDE OF GOOD PRACTICES FOR REGIONS WITHOUT

FLAVESCENCE DORÉE
1
Picture on the front page : Vineyard infected with Flavescence dorée (Institute of Agriculture and Tourism – abbrev. IPTPO)
F L AV E S C E N C E D O R É E
EPIDEMIOLOGY – A BRIEF SCIENTIFIC OVERVIEW
Flavescence dorée is a quarantine grapevine disease indexed on A2 EPPO list (2000/29/EC)2

and present in numerous winegrowing regions of Europe with a tendency of further spreading.
Flavescence dorée (hereafter: FD) is a complex disease that integrates three essential elements
present either in a vineyard or in the surrounding environment: causal agent – phytoplasma Ca.
Phytoplasma vitis (hereafter: FDp), insect vectors that transmit the disease among host plants
(Scaphoideus titanus, Dictyophara europaea, Oncopsis alni, Orientus ishidae) and host plants that
serve as a phytoplasma reservoir (Vitis spp., Alnus glutinosa, Clematis vitalba) (Fig. 1) (Schvester et
al., 1963; Caudwell et al. 1994; Maixner et al., 2000; Filippin et al., 2009; Lessio et al., 2016). The
number of newly infected vines in a current year may be significantly different as a result of the
vector-host specimen present in the winegrowing area and not all previously listed vectors and hosts
plants are able to induce FD outbreaks according to EFSA evaluation (EFSA PHL, 2016).
Highly Only
significant! occasional.
Collective
management of S.
Significant for
epidemical disease
titanus and
spread in infected grapevines
winegrowing areas. is fundamental!
Fig. 1: FD epidemiology – vectors and host plants. (Graphics: K. Diklić, IPTPO).
2 Directive 2000/29/EC will be repealed on 14 December 2019 and will be replaced by Regulation (EU) 2016/2031 of the European
Parliament and of the Council concerning protective measures against pests of plants.
Page 1
FD phytoplasma may be transmitted from several host plants on grapevine with vectors other than
S. titanus, that are present in the surrounding environment but these transmissions are not relevant
for FD outbreaks. Why other vectors and hosts are not involved in epidemic spread?
The frequency of FDp transmission from Alnus glutinosa, Clematis vitalba, Ailanthus altissima with
vector species Dictyophara europaea and Oncopsis alni to grapevine is low and occasional because
these vectors are not as frequent feeders on grapevine as S. titanus (Maixner et al., 2000; Arnaud et
al., 2007; Filippin et al., 2009). The first introduction of FDp in the vineyard with one of the
listed ‘secondary’ vectors, in winegrowing areas where previous FD established on grapevine were
not recorded, may be the first step of further epidemical FD spread by S. titanus (Fig. 2).
Interaction grapevine – FDp – S. titanus is essential for FD outbreaks, why? Outbreaks
of FD in producing vineyards are related to the presence of both FDp in Vitis spp. and high
populations of vector S. titanus. The insect vector S. titanus has a feeding preference for Vitis species
where may be present from larval (May) to adult development stage (October) and transmit the FDp
by feeding from larval L4 to adult development stage (Chuche et al., 2014) (Fig. 2). Current
experiences with FD management indicate that an increase in disease incidence from 10-fold (Smith
et al., 1997) up to 40-fold (Prezelj et al., 2012) may occur if no control measures had been
implemented in S. titanus control. Other potential vectors in FD outbreaks? Lately it has
been evidenced that Orientus ishidae is rapidly spreading in Europe (Lessio et al., 2016) and a
hypothesis that it may have a potential impact on FD outbreaks in some areas is introduced but
more profound scientific data are required.
S. titanus acquires FDp

from the infected
grapevine and
transmits it further.
S. titanus transmits
FDp to grapevine.
No visible
Symptoms development.
symptoms.
Fig. 2: Scaphoideus titanus and FDp – inoculation and transmission (Year n – Inoculation of FDp and
lack of visible symptoms; Year n+1 – symptoms development and potential transmission of FDp to
new vines). (Chuche, 2010)
Page 2
DISTRIBUTION IN EUROPE
DISTRIBUTION ACCORDING TO EFSA3
Flavescence dorée phytoplasma is widely present in winegrowing regions of France, Italy, Slovenia
and Serbia. Restricted FD distribution is present in Austria, Croatia, Hungary, Portugal, Spain and
Switzerland (Fig. 3).
Fig. 3: Distribution of FD and S. titanus in Europe (Abbreviations: S. titanus , FD presence ,

Vineyards Corine Landov 2012 ). (EFSA PHL, 2016)
3EFSA Panel on Plant Health (PLH), Michael Jeger, Claude Bragard, David Caffier, Thierry Candresse, Elisavet Chatzivassiliou, Katharina
Dehnen-Schmutz, Gianni Gilioli, Josep Anton Jaques Miret, Alan MacLeod, Maria Navajas Navarro, Björn Niere, Stephen Parnell, Roel
Potting, Trond Rafoss, Vittorio Rossi, Gregor Urek, Ariena Van Bruggen, Wopke Van Der Werf, Jonathan West, Stephan Winter,
Domenico Bosco, Xavier Foissac, Gudrun Strauss, Gabor Hollo, Olaf Mosbach-Schulz, Jean-Claude Grégoire (2016). Risk to plant health
of Flavescence dorée for the EU territory. EFSA Journal 2016;14(12):4603, 83 pp.
http://onlinelibrary.wiley.com/doi/10.2903/j.efsa.2016.4603/full
Page 3
SYMPTOMS
Key symptoms are described according to experiences from Piemonte region (Italy)
(www.regione.piemonte.it) and Istria region (Croatia). Monitoring of symptoms development may
be conducted during different parts of vegetation but later in summer, typical symptoms are more
easily visible. Symptom monitoring and labelling of symptomatic vines needs to be done prior
harvest, since grapes and leaves in bunch zone are removed during this operation.
SPRING:
(http://www.regione.piemonte.it)
- Shoots on the fruiting cane have a reduced growth (reduced number of internodes) (Fig. 4),
- Shoots on the fruiting cane develop shortened internodes with zig-zag growth,
- Leaf surface is reduced (Fig. 4),
- Blistering of leaf blade as a consequence of reduced leaf veins development,
- Desiccations development on shoots from apical toward basal part (Fig. 5),
- Slight leaf roll downwards (Fig. 5),
- Insertion of leaf blade on petiole is more ‘sharp’,
- Premature leaf fall,
- Browning of the inner part of the cortex on symptomatic fruiting canes of diseased vines.
A B C
Fig. 4: Reduced shoot and leaf area growth (A, B) and leaf yellowing (C) on cv Istrian Malvasia (K.
Diklić, IPTPO)
A B C
Fig. 5: Symptomatic shoots of cv Cortese (A) (M. Gily, SIVE), apical shoot necrosis (B) and leaf colour
aberration on cv Istrian Malvasia (C) (K. Diklić, IPTPO)
Page 4
SUMMER:
(www.regione.piemonte.it)
a) Early summer
- Shoots with reduced growth (stunted growth during spring),
- Leaf colour aberrations – reddening (red varieties) or yellowing (white varieties) : near leaf
veins, leaf sections delimitated by veins or complete leaf chlorosis (Fig. 6),
- Leaf roll downwards (visible only in some cultivars, example Chardonnay),
- Premature leaf fall (‘defoliation’ caused by FDp) due to leaf detachment from petiole
(detachment of leaf blade without petiole, the petioles sometimes remain on the shoot) (Fig.
6),
- Desiccation of inflorescences approximately after flowering,
- Desiccation of berries approximately after fruit set or later during berry softening (Fig. 6),
- Complete lack of grape bunches on grapevine (100% yield loss).
b) Late summer
- Shoot gummosis and lack of lignification on several/all shoots developed on the fruiting cane
(Fig. 6),
- Shoot growth is not straight due to gummosis consistency, vines assemble “umbrella-like”
growth,
- Thickening of leaf blade due to sugar accumulation – symptomatic leaf is brittle and cracks
when folded in hand.
Fig. 6: Symptoms on Istrian Malvasia: lack of cane lignification, grape bunches desiccation at fruit
set, leaves yellowing and ‘defoliation’ caused by FDp. (K. Diklić, IPTPO)
Page 5
L E A F SY M P TO M S – W H I T E G R A P E V I N E VA R I E T I E S
A B C D
Fig. 7: Leaf symptoms on different varieties: A – Chardonnay (K. Diklić, IPTPO), B – Chardonnay (IFV, South-West), C – Pinot blanc (K. Diklić, IPTPO), D – Muscat blanc (K. Diklić, IPTPO)
A B C D E
Fig. 8: Leaf symptoms on different varieties: A, B – Istrian Malvasia (K. Diklić, IPTPO), C – Len de l'El (IFV, South-West), D - Sauvignon blanc (IFV, South-West), E - Muscadelle (IFV, South-West)
A B C D
Fig. 9: A – Leaf symptoms Moscato (M. Gily, SIVE), B, C – premature leaf fall – detachment of blade and petiole (K. Diklić, IPTPO), D – premature leaf fall on a shoot (K. Diklić, IPTPO)
L E A F SY M P TO M S – R E D G R A P E V I N E VA R I E T I E S
A B C D E
Fig. 10: Leaf symptoms on varieties: A – Cabernet Sauvignon (K. Diklić,IPTPO),B – Duras (IFV, South-West),C – Syrah (IFV, South-West),D – Gamay (IFV, South-West),E – Plavina (K. Diklić,IPTPO)
Fig. 11: Different colour aberrations on leaves of variety Teran (K. Diklić, IPTPO)
A B C D E
Fig. 12: Leaf symptoms on varieties: A – Barbera (M. Gily, SIVE), B – Fer Servadou (IFV, South-West), C – Gringolino (M. Gily, SIVE), D – Merlot (ERSA), E – Carmenere (ERSA)
Page 7
SYMPTOMS ON GRAPEVINE INFLORESCENCES AND BERRIES
A B C
Fig. 13: Desiccation of grapevine inflorescence on cv Istrian Malvasia (A, B) (K. Diklić, IPTPO), desiccation of grapevine bunch post verasion on cv Moscato (C) (M. Gily, SIVE)
A B C D
Fig. 14: Desiccation of grapevine bunch: after fruit setting (A, B), after verasion (C, D) on cv Istrian Malvasia (K. Diklić, IPTPO)
Page 8
S Y M P T O M S - D I S E A S E I M PA C T O N Y I E L D
In the first years of FD establishment in a new area winegrowers misinterpret FD symptoms, but significant yield losses (Fig. 15) are always a sign that
actions must be taken!
Fig. 15: Variety Muscat blanc – asymptomatic (blue circle) and symptomatic (red circle) vines
Symptomatic: complete lack of bunches, leaves colour aberrations (yellowing), leaf roll downwards, shortened shoot growth, vigour significantly reduced (K. Diklić, IPTPO)
Page 9
FRUITING CANE SYMPTOMS
A B C D
Fig. 16: Symptoms on fruiting canes of different varieties: A – ‘curled’ cane growth due to cane gummosis, B – not lignified cane, C - D black pustules on cane (K. Diklić, IPTPO)
A B C D
Fig. 17: Symptoms on fruiting canes of different varieties: A – C ‘curled’ cane growth due to cane gummosis consistence, D – not lignified canes (K. Diklić, IPTPO)
Page 10
S Y M P T O M S S I M I L A R T O F L AV E S C E N C E D O R É E
SPRING
Shoots reduced growth in spring may be a consequence of early spring eriophid bud mite activity, which is more usual in colder springs when vegetation is
developing slower than usual. If shoot growth is reduced due to mite activity, there are visible scare tissues on shoots and leaves. Shoot growth may be
reduced also as a result of vine imbalance (over cropping), boron or zinc deficiency, frost damage, poor herbicide application (Walton et al., 2009).
A B C D
Fig. 18: Eriophid bud mites damage: A – B stunted shoot growth in spring; C – D symptom similar to leaf blistering caused by FDp, but in the case of mites on
the leaf underside (below the blistering) there are white mite colonies (K. Diklić, IPTPO)
Fig. 19 (Left): Herbicide applications during late autumn or early spring may cause
leaves distortions and stunted shoot growth if the treatment is not applied correctly (K.
Diklić, IPTPO)
Fig. 20 (Right): Leaves discoloration

on variety Barbera due to frost
damage during spring. (M. Gily, SIVE)
Page 11
SUMMER
Fig. 21 (Left): Magnesium deficiency

on cv Chardonnay (A) and Istrian
Malvasia (B), iron deficiency on cv
Istrian Malvasia (C) may be confused
with foliar discolorations caused by
FDp. (K. Diklić, IPTPO)
A B C
Fig. 22: Grapevine bunch – physiological disorders on inflorescences of different varieties: poor fruit set due to unfavourable conditions during flowering (K. Diklić, IPTPO)
Fig. 23: Grapevine bunch – physiological disorders: bunch stem necrosis of different varieties (K. Diklić, IPTPO)
Page 12
A B C D
Fig. 24: Grapevine bunch: A – B grapevine mites on bunch during fruit setting (K. Diklić, IPTPO), C – D grapevine bunch infected with Plasmopara viticola (C – Terra e vita, D – ARSIA)
A B C D
Fig. 25: Leaf symptoms of grapevine viruses that belong to the complex of Grapevine leafroll (A – C: red varieties, D – Chardonnay) (K. Diklić, IPTPO)
A B C D
Fig. 26: Leaf symptoms of Nepovirus grapevine virus species (A – chromogenic virus strain on cv Moscato giallo; B – distorting virus strain on cv Pagadebit) and Grapevine Pinot gris virus (C, D
– cv Refošk) prior flowering (K. Diklić, IPTPO)
Page 13
Fig. 27: Esca disease symptoms on cv Istrian Malvasia (left) and Teran (right): not lignified Fig. 28: Stictocephala bisonia damage on grapevine shoot leads to leaf discolorations
shoots, leaf symptoms typical for Esca, grape desiccation (K. Diklić, IPTPO) (ERSA)
S Y M P T O M S O F F L AV E S C E N C E D O R É E O N O T H E R H O S T S
A B C D
Fig. 29: A - FD symptomatic (left) and asymptomatic rootstock leaves (K. Diklić, IPTPO), B – FD infected but asymptomatic rootstock (K. Diklić, IPTPO), C - wild symptomatic Vitis spp. (K. Diklić,
IPTPO), D - SO4 rootstock with symptoms (IFV, South-West)
Page 14
V E C T O R S C A P H O I D E U S T I TA N U S
Vector S. titanus induces no significant symptoms on grapevine when feeding on leaves, therefore vector presence is observed by monitoring larvae on the
downside part of grapevine leaves or adult stages on yellow sticky traps positioned within the vineyard (Fig. 30 – 31). S. titanus larvae have five development stages
of a size range from 1.8 (L1) to 5.2 mm (L5) and they are mainly located on basal shoot leaves on the fruiting canes or leaves of trunk suckers (Cara et al., 2013;
Trivellone et al., 2015). Distribution of S. titanus larval stages is observed predominantly in grapevine canopy and inter-row vegetation with different density
populations during the vegetative season, but different plant species present in the inter-row area or area close to the vineyard plot may influence the larval spatio-
temporal distribution (Trivellone et al., 2013). Larval development stages don’t have wings and presumably are not able to disperse on long distances like adults,
while according to some researches adult development stages of S. titanus disperse from wild grapevines to vineyards mainly within 30m, there are also evidences of
adult dispersal up to 330 m from areas where wild vines are present (Lessio et al., 2014).
A B C D
Fig. 30 S. titanus larvae monitoring: A - S. titanus larvae development stage L1 (IFV South-West), B - S. titanus larvae (N. Burghardt, EKU Eger), C - S. titanus larvae (P. Rózsahegyi, EKU Eger), D
– S. titanus larvae (K. Diklić, IPTPO)
A B C D
Fig. 31 S. titanus adult monitoring: A – Analysis of captured S. titanus on the yellow sticky trap (K. Diklić, IPTPO), B – S. titanus on yellow sticky trap (N. Burghardt, EKU Eger), C – monitoring
with entomological net (N. Burghardt, EKU Eger), D – S. titanus on fabric of the entomological net (N. Burghardt, EKU Eger)
Page 15
PROCEEDINGS IN CASE OF FD INTRODUCTION

WINEGROWERS
In cases of FD introduction in new areas, previously marked as uninfected, control and eradication
measures are applied in correspondence with European and National and/or regional laws. Report
of symptomatic plants potentially infected with FDp is obligatory in the winegrowing regions of EU
as a result of FD quarantine status. What should be done in the case of suspected
introduction of FD in a new area?
1. Notification of potential introduction of FDp in new areas needs to be communicated
to national or regional institutions included in the winegrowing sector:
a) Phytosanitary services,
b) Regional advisory services,
c) Research and/or technical institutes,
d) Organizations of winegrowers, etc.
2. Collection and analysis of grapevine samples for FDp presence in collaboration with
the national or regional authorized organization.
3. Information of local producers in case of FD outbreak in a new area in order to signal all
potential foci in the area and organize synchronized disease eradication is implemented
by official organizations.
4. Education of wine industry stakeholders. Request of information on measures

obligatory to implement in case of FD outbreak in a new area (defined by national or regional
law) in order to eradicate the disease and prevent the further spread.
CRUCIAL PRACTICES IN FD PREVENTION

ACTIVITIES IN AREAS WITHOUT FD
NURSERY PRODUCERS
Production of planting material according to EPPO recommendations 4 (for further information see:
EPPO – PP2/023(1) Grapevine) in nursery production with special focus on:
1. Control of symptoms development in mother plants (remember: rootstocks mainly don’t
develop FD symptoms),
2. Implementation of diagnostic tools to control potential FDp infections (for further
information see: EPPO4 - PM 7/079 (2) Grapevine flavescence dorée phytoplasma),
3. Import of scion and rootstock material from areas with present FDp for production of
planting material must be avoided or when realized additional measures like Hot water
treatment are advised,
4. Implementation of Hot water treatment protocols that limit the spread of FDp (EPPO4 –
PM10/018 (1) Hot water treatment of grapevine to control Grapevine flavescence dorée
phytoplasma),
5. Monitoring and control of Scaphoideus titanus larval and adult stages is fundamental as a
preventive measure even in areas where FDp is not present.
WINEGROWERS
Implementation of practices in viticultural production in order to avoid FDp introduction needs to be
oriented especially on (EPPO – PP2/023(1) Grapevine; EPPO – Data sheet on Grapevine flavescence
dorée phytoplasma4):
1. Planting certified (healthy) planting material,
2. Control of symptoms development in Vitis vinifera varieties (remember: rootstocks mainly
don’t develop FD symptoms) with further molecular analysis of planting material for FDp
presence,
3. Monitoring and control of Scaphoideus titanus larval and adult stages is fundamental as a
preventive measure even in areas where FDp is not present. Highly significant in areas
approximate to winegrowing regions with confirmed FDp presence.
WHY PREVENTION IS CRUCIAL?
According to EFSA PHL (2016) there are three mechanisms of FDp spreading: (1) trade and
movement of infected propagative material, (2) infected vectors flying from adjacent spatial units,
transported on plants for planting or hitchhiking in vehicles, (3) transfer from the wild compartment
(infected Alnus and Clematis). Furthermore, experts evaluated that infected planting material
and S. titanus are significant for introduction of FDp in new areas (EFSA PHL, 2016) and
the previously listed practices are crucial in preventive disease management and limitation of
further FD spread.
4
Additional information is available on EPPO website : https://gd.eppo.int/taxon/PHYP64/documents
Page 17
WINEGROWERS AND NURSERY PRODUCERS:

AC T I V I T I ES TO I M P L E M E N T
COLLECTIVE DISEASE MANAGEMENT5
1. Monitoring of leafhopper S. titanus larvae during May – June is
fundamental if regional recommendations for S. titanus insecticide
treatments are not available.
Monitoring for determination of larval stage (L1 – L5) and distribution
of S. titanus in vineyard needs to be oriented to (Fig. A):
a) Vector monitoring with the beating tray or visual control of leaves
underside positioned on basal grapevine shoots internodes
(shoots that develop from fruiting canes) – approximately 100
A
leaves/ha,
b) Visual control of leaves underside located on grapevine suckers
(basal shoots on the trunk).
2. Activities prior insecticide treatments:
a) Removal of grapevine suckers prior insecticide treatment is
needed because if not implemented, insecticide treatments need
to cover the surface of grapevine suckers since S. titanus larvae are
found there also, B
b) Mulching of cover-crops if plants are flowering in order to prevent
bees poisoning,
c) Notification of bee keepers or associations prior insecticide
applications in order to prevent bees poisoning.
3. Insecticide treatment for S. titanus control (oriented to larvae)
prior potential FDp transmission (to be implemented in both
infected and « buffer » zones): C
a) Organic production: (1) treatment of L1-L2 larvae in the beginning
of June (prior flowering), (2) treatment of L3 larvae in mid-June
(post flowering),
b) Integrated production6: (1) treatment of L3 larvae in mid-June
(post flowering), (2) treatment 2-3 weeks after (adaptation
according to national regulations is needed).
Beware of pollinators’ populations by not applying insecticide
treatments during flowering and postponing insecticide
treatments later in the evening!
4. In the period from July to harvest (after the first treatment
oriented to larval development stages) elimination of parts of
symptomatic vines may be applied to prevent potential
transmission of FDp to new vines in eventual remaining vector
populations:
a) removal of shoots (Fig. C),
b) basal cut on the trunk.
5 Pictures: K. Diklić, Institute of Agriculture and Tourism (Croatia)
6 Directive 2009/128/EC
Page 18
Complete vine rouging usually is implemented post-harvest or

during winter. If this approach is not applicable, marking
symptomatic vines with spray or highly-visual tapes is possible
in order to recognize infected vines during winter.
5. Monitoring of S. titanus (adult development stage) with
yellow sticky traps (YST) from July until harvest to verify if
there are remaining vector populations. If adults of S. titanus
are recorded, an additional third insecticide treatment may be D
applied (Fig. D, E). Consider possible differences in the
population level of S. titanus on vineyard border and in the
central part of the vineyard (Fig F).
6. Application of a third insecticide treatment when required
(adults of S. titanus identified on YST), approximately 2 – 3
weeks after the second treatment, is later in the season and
potential residues in wine need to be considered and avoided.
Application of the last treatment needs to be done according to E
the PPPs recommendations.
7. Monitoring for eventual outbreaks of grapevine mites (visual
leaf monitoring) that may be present as a result of imbalance
due to broad spectrum insecticide treatments.
8. Activities in the period between post-harvest
(September/October) and May: F
a) Rouging of singular symptomatic vines with root to prevent
infected (and prevalently asymptomatic) rootstock re-growth G
(less than 20% symptomatic vines) (Fig. G, H),
b) Complete extirpation of symptomatic vineyards (more than 20%
symptomatic vines),
c) Pruning debris management (mulching, etc.) in order to reduce
vector populations: vector S. titanus deposits eggs under the bark
of the 2-year-old cane (egg hatching occurs usually not prior May
and this procedure may be implemented in any period prior that).7
REMEMBER: Registration of applied insecticide treatments and
recycling PPPs containers is obligatory.8
H
INSECTICIDE CHOICE: verify PPPs available on the national level for
S. titanus management in integrated or organic grapevine
production. Some PPPs applied for S. titanus control are also
efficient for Lobesia botrana control.
Collaboration and commitment is essential

in effective disease management!
7 Pictures: K. Diklić, Institute of Agriculture and Tourism (Croatia)

8 http://www.ecpa.eu/regulatory-policy-topics
Page 19
M O R E I N F O R M AT I O N
L I T E R AT U R E
1. Arnaud G., Malembic-Maher S., Salar P., Bonnet P., Maixner M., Marcone C., Boudon-Padieu
E., Foissac X. (2007). Multilocus sequence typing confirms the close genetic inter-relatedness
between three distinct flavescence dorée phytoplasma strain clusters and group 16SrV
phytoplasmas infecting grapevine and alder in Europe. Applied and Environmental
Microbiology, 73, 4001-4010.
2. Cara C., Trivellone V., Linder C., Junkert J., Jermini M. (2013). Influence de la gestion des
repousses du tronc et du bois de taille sur les densités de Scaphoideus titanus. Revue Suisse
Viticulture, Arboriculture, Horticulture, 45(2), 114-119.
3. Caudwell A., Larrue J., Tassart V. (1994). Ability of grapevine rootstocks varieties to transmit
flavescence doree. Study of the case of 3309 C and Fercal. Agronomie (France).
4. Chuche J. (2010). Comportement de Scaphoideus titanus, conséquences spatiales et
démographiques. Doctoral dissertation, Bordeaux 2, France.
5. Chuche J., Thiéry D. (2014). Biology and ecology of the Flavescence dorée vector Scaphoideus
titanus: a review. Agron. Sustain. Dev., 34, 381-403.
6. EFSA PHL (2016). Jeger M, Bragard C, Caffier D, Candresse T, Chatzivassiliou E, Dehnen-
Schmutz K, Gilioli G, Jaques Miret JA, MacLeod A, Navajas Navarro M, Niere B, Parnell S,
Potting R, Rafoss T, Urek G, Rossi V, Van Bruggen A, Van Der Werf W, West J, Winter S, Bosco
D, Foissac X, Strauss G, Hollo G, Mosbach-Schulz O and Grégoire J-C. Scientific opinion on the
risk to plant health of Flavescence dorée for the EU territory. EFSA Journal, 14(12):4603, 83
pp.
7. Filippin L., Jović J., Cvrković T., Forte V., Clair D. Toševski I., Boudon-Padieu E., Borgo M.,
Angelini E. (2009). Molecular characteristics of phytoplasmas associated with Flavescence
doree in clematis and grapevine and preliminary results on the role of Dictyophara europaea
as a vector. Plant Pathology, 58, 826–837.
8. Lessio F., Tota F., Alma A. (2014). Tracking the dispersion of Scaphoideus titanus Ball
(Hemiptera: Cicadellidae) from wild to cultivated grapevine: use of a novel mark-capture
technique. Bulletin of Entomological Research, 104, 432-443.
9. Lessio F., Picciau L., Gonella E., Mandrioli M., Tota F., Alma A. (2016). The mosaic leafhopper
Orientus ishidae: host plants, spatial distribution, infectivity, and transmission of 16SrV
phytoplasma to vines. Bulletin of Insectology, 69, 277-289.
10. Maixner M., Reinert W., Darimont H. (2000). Transmission of grapevine yellows by Oncopsis
alni (Schrank) (Auchenorrhyncha: Macropsinae). Vitis, 39, 83–84.
11. Papura D., Burban C., van Helden M., Giresse X., Nusillard B., Guillemaud T., Kerdelhue C.
(2012). Microsatellite and mitochondrial Data Provide Evidence for a Single Major
Introduction for the Neartic Leafhopper Scaphoideus titanus in Europe. PLoS ONE, 7(5),
e36882.
12. Prezelj N., Nikolić P., Gruden K., Ravnikar M., Dermastia M. (2012). Spatiotemporal
distribution of flavescence dorée phytoplasma in grapevine. Plant pathology, 62:4, 760-766.
13. Schvester D., Carle P., Moutous G. (1963). Transmission de la flavescence dorée de la vigne
par Scaphoideus littoralis Ball. Annales des Epiphyties, 14, 175–198.
14. Smith I.M., McNamara D.G., Scott P.R., Holderness M., eds. (1997). Quarantine Pests for
Europe, 2nd edition. CAB International, Wallingford, UK, pp. 1013–1021.
Page 20
15. Steffek R., Reisenzein H., Zeisner N. (2007). Analysis of the pest risk from Grapevine
flavescence dorée phytoplasma to Austrian viticulture. Bulletin EPPO 37, 191-203.
16. Trivellone V., Corrado C., Jermini M. (2015). Répartition spatio-temporelle de la cicadelle
Scaphoideus titanus Ball dans l’agroécosystème viticole. Revue suisse Viticulture,
Arboriculture, Horticulture, 47(4), 216-222.
17. Trivellone V., Jermini M., Linder C., Cara C., Delabays N., Baumgärtner J. (2013). Rôle de la
flore du vignoble sur la distribution de Scaphoideus titanus. Revue suisse Viticulture,
Arboriculture, Horticulture, 45(4), 222-228.
18. Walton V., Skinkis P., Dreves A., Kaiser C., Renquist S., Castagnoli S., Hilton R. (2009).
Grapevine growth distortions. A guide to identifying symptoms.
https://catalog.extension.oregonstate.edu/sites/catalog/files/project/pdf/em8975.pdf
19. http://agroambiente.info.arsia.toscana.it/arsia/arsia14?ae5Diagnosi=si&IDColtura=2&IDSche
daFito=1
20. http://www.ersa.fvg.it/istituzionale/servizio-fitosanitario-regionale/organismi/flavescenza-
dorata-e-altri-giallumi-della-vite/plonearticlemultipage.2007-05-30.4633965054/cicalina-
bufalo-stictocephala-bisonia
21. http://www.ersa.fvg.it/prova/guida-alla-diagnosi/flavescenza-dorata-e-altri-giallumi-della-
vite/sintomi-specifici-per-singola-varieta/vitigni-neri/plonearticlemultipage.2007-06-
05.2647684871/sintomi-sulle-foglie
22. http://www.regione.piemonte.it/agri/area_tecnico_scientifica/settore_fitosanitario/vigilanza
/flavescenza.htm
23. http://www.terraevita.it/vite-proteggere-il-grappolo/
EPPO
24. PM1/002(25) EPPO A1 and A2 Lists of pests recommended for regulation as quarantine
pests (2016)
25. PM10/018(1) Hot water treatment of grapevine to control Grapevine flavescence dorée
phytoplasma
26. PM4/008(2) Pathogen-tested material of grapevine varieties and rootstocks
27. PM7/079(2) Grapevine flavescence dorée phytoplasma
28. PP2/023(1) Grapevine
WINETWORK KNOWLEDGE RESERVOIR

http://www.winetwork-data.eu/en/hr/default.asp
Page 21
C O N TA C T
FACILITATOR AGENTS
Kristina Diklić - dkristina@iptpo.hr

Institute of Agriculture and Tourism (Croatia)
Fanny Prezman – fanny.prezman@vignevin.com

Institut Français de la Vigne et du Vin – South-West (France)
Céline Abidon - celine.abidon@vignevin.com

Institut Français de la Vigne et du Vin – Alsace (France)
Natasa Burghardt – burghardt.natasa@ektf.hu

Eszterházy Károly University of Applied Sciences (Hungary)
Tabitha Kellerer - tabitha.Kellerer@dlr.rlp.de

Dienstleistungszentren Ländlicher Raum (Germany)
Constanze Mesca - constanze.mesca@dlr.rlp.de

Dienstleistungszentren Ländlicher Raum (Germany)
Paula Aldeanueva Potel – paldeanueva@feuga.es

Fundación Empresa-Universidad Gallega (Spain)
María Jesús Fanjul Alonso – mjfanjul@xunta.es

The Instituto Galego da Calidade Alimentaria (Spain)
Diego López Llaria – dlopezllaria@gmail.com

Fundación Empresa-Universidad Gallega (Spain)
Igor Gonçalves – Igor.goncalves@advid.pt

Associação para o Desenvolvimento da Viticultura Duriense (Portugal)
Maurizio Gily – maurizio@gily.it

Società Italiana Viticoltura Enologia (Italy)
Cristina Micheloni – cristina.micheloni@gmail.com

Società Italiana Viticoltura Enologia (Italy)
Page 22
APPENDIX 5: FLAVESCENCE DORÉE: HOW TO MANAGE THE
DISEASE WITH MORE PRECISION
www.winetwork.eu
16/27
FLAVESCENCE DORÉE : HOW TO MANAGE

THE DISEASE WITH MORE PRECISION ?
In the regions already infected by Flavescence Dorée disease

F L AV E S C E N C E D O R É E : H O W T O M A N A G E W I T H
MORE PRECISION?
Flavescence dorée (FD) is the most important and destructive phytoplasma disease of grapevines. FD
induce severe impacts, including reduced vitality of vines, yield reductions and reduced wine quality.
Without control measures, the disease spreads rapidly, affecting up to the totality of vines in a few years.
Despite mandatory control in Europe for this disease, it is still spreading and need permanent monitoring to
detect new infected areas.
A good management of FD go through a combination of methods working both on the vector, Scaphoideus
titanus and on the disease, once grapevine are contaminated by the phytoplasma. In infected regions,
management of the vector and of the disease go hand in hand. The conscientious application of the
methods described in this datasheet (vector, vineyard and wild vines management and when needed, hot
water treatment) are essential to control FD in a territory and to limit its spread.
VECTOR MANAGEMENT
1 - T I M I N G F O R S P R AY I N G : A K E Y FA C T O R
Application time is the key of success in managing Scaphoideus titanus population in vineyard. The
leafhopper transmits FD phytoplasma from one grapevine to another very quickly. In order to slow
disease’s propagation, a good control of the vector and management of the vineyard at a large scale are
necessary.
In order to apply insecticides against the leafhopper at the good timing, several method exist to determine
the best application date:
1- Emerging cages: these cages are a very effective tool to determine when Scaphoideus titanus first
hatching appears. 2 years old wood, preferred for egg laying, from a plot where Scaphoideus titanus
presence was detected in the previous year are kept in a cage and hatching moment is monitored
every day. A sticky trap is put inside the cage in order to capture the larvae as soon as they hatch.
When date of first hatching is determined the effective date for insecticides treatments are positioned
one month later.
2- Modelling: decision support systems exists and are able to predict in vineyard (with its local conditions)
pests and diseases, including Scaphoideus titanus. One model currently in development can predict
when each stage of Scaphoideus titanus nymphs will appear according to specific climatic conditions
(this implies to have a weather station close to the plot). Model is based on observations made the
years before and historical and daily temperatures. In order to provide a more accurate possible real
observation data on stages need to be informed.
3- Monitoring of nymphal development stages, on leaves is requested to choose the right time for the first
treatment, but cannot usually be made by winegrowers and requires trained technicians. Data on
nymphs repartition can allow to divide a region into macroclimatic areas, monitor the insect and
suggest the best time for spraying in each area, and even suggesting a different timing depending on
the kind of product used. Visual control need to be done on leaf underside and on grapevine basal
shoots (suckers) and basal leaves from mid-May to August in the early morning avoiding to move
Page 1
excessively the vegetation. There are 5 successive nymphs stages from hatching to adult, nymphs are
identifiable thanks to two symmetrical black points in dorsolateral position at abdomen posterior end
(nymphs can be confused with Phlogottetix Cyclops who has also two black points on the second-last
segment of abdomen, see picture below). These points are visible to the naked eye from L2. The
nymph, when disturbed, show a typical behaviour: it tends to jump away. This behaviour can be used to
discriminate S.titanus nymphs to other leafhopper juvenile forms that should be present at the same time
on the grapevine leaves, such as Empoasca vitis (when disturbed show to move laterally on the leaf
surface) and Zygina rhamni (when disturbed show to move along a straight line on the leaf surface).
4- Monitoring adult with traps: yellow sticky traps can be set up in vine-plot and can be monitored by
growers themselves if properly instructed, or by specialists. A good tool to find again the traps for
people who control many of them is to use GPS localized shots. Traps will help to decide when apply
adulticide treatment. Adult flights have been often observed from the beginning of July to later than
expected (October). The late control of vector leafhopper population is important to decide the
opportunity of an additional insecticide against adults.
Sticky traps are yellow because Scaphoideus titanus adults are more attracted by this colour. Sticky
traps need to be set up when L4 to L5 nymphs are observed on the plot. Sticky traps can be used on
plots with a strong FD history in order to monitor the flight and on plots free from FD to prevent
infection. On the plot with a strong S.tianus populations history one sticky trap can usefully help to know
when adults are flying, for a plot free from FD vector, several traps should be hanged in the plot, with
a focus on the edges.
Traps need to be hanged inside the foliage, the as close as possible to vegetation, where leafhoppers
are present, usually they are placed on the height of 1,5 m between two wireS.the number of traps
per plot is usually different according to the size of the plot and number of surrounding vineyard or
areas with wild Vitis species. Usually, 5-6 sticky traps/ha are placed and checked every 7 days.
How to recognize the adult?

Another method to monitor adults is the beating method. It consist of the use of an entomological
umbrella made of white cloth which is positioned under the vine canopy to collect S.titanus individuals
that fall in after vine shaking (then they are counted). Efficiency of this method is limited but can
present better efficiency in the first two-three days of adult appearance because they are less mobile.
After these days the adults tend to fly away flying up to few kilometres. So, it is more probably that
they are attracted from the yellow colour of traps. Another alternative method is suckering of
leafhopper with D-Vac.
Scaphoideus titanus adult size range from 4,8 to 5,8 mm, has brown colour and stripes on the head
(from 1 to 3 according gender). Scaphoideus titanus adult can be mistaken with Phlogotetix.
Page 2
Mistakeable species
Phlogotettix cyclops nymph (INRA Bordeaux) Phlogotettix cyclops adult (IPTPO)
Hyalesthes obsoletus (hemiptera-databases.org) Oncopsis alni (INRA Bordeaux)
Empoasca vitis nymph (IFV South-West) Emposasca vitis adult (IFV South-West)
Dictyophara europaea (IPTPO)
Page 3
2- T R E A T M E N T S T R A T E G I E S A N D A P P L I C A T I O N
According to the region and even to the country, several treatment strategies exist. One example is given
here. The first treatment is the most important, eggs hatching need to be monitored (with one of the
methods described above) in order to know when positioning first treatment. The first treatment need to be
positioned one month after hatching because when nymphs first feed on an infected grapevine there is an
incubation time of one month for the nymphs begin infectious and can transmit the phytoplasma again.
Strategy with 3 treatments

T1: One month after hatching
T2: End of remanance of T1 product, objective to cover end of hatching
T3: On adults
Or
T1: One month after hatching or prior L4-L5
T2: On adults
T3: On adults late summer or after harvest depending on Sc.t population
Strategy with 2 treatments (low population of Sc. t. and no infected grapevines):

T1: On nymphs
T2: On adults
Strategy with extra treatments (high population of Sc.t in the previous years)
T0: Extra treatment on L1 in middle of May, before flowering
T1: On nymphs
T2: End of remanance of T1 product, objective to cover end of hatching
T3: On adults
Page 4
Insecticide treatment application need to cover all grapevine leaf area (fruiting canes and basal suckers),
from the basis to the top. Indeed, Scaphoideus titanus nymphs are often located close to the first leaves of
the trunk and on the suckers, mainly because eggs were laid under 2 years-old bark. Prior to treatment
application it is recommended to remove basal shoots and spray need to cover trunk and shoots.
Application need to be done respecting product’s recommendation and at the proper timing.
3- PRODUCTS (CONVENTIONAL AND ORGANIC)
CONVENTIONAL CHEMICAL CONTROL OF FD VECTOR SCAPHOIDEUS TITANUS
ACTIVE INGREDIENT USABLE IN NURSERY NBR OF MAX USE AUTHORIZED IN

TREATEMENT VINEYARD IN
/ YEAR
Acetamiprid Italy 3 Italy
Acrinathrin France, Italy, Portugal 1-3 France, Italy, Portugal
Alpha-Cypermethrin France, Italy, Hungary, 2-3 France, Italy, Hungary, Portual
Portugal
Alphamethrin France France
Azadirachtin Croatia 2 Italy, Croatia, Italy, Portugal

Beta-Cylfuthrin France, Italy 2-3 France, Italy, Hungary
Buprofezin Italy 1 Italy
Chlorantraniliprole-Thiamethoxam France, Hungary, Portugal 1 France, Hungary, Italy,
Portugal
Chlorpyrifos-ethyl France, Italy 3 France, Italy
Chlorpyrifos-Cypermethrin France, Croatia, Hungary, 1-2 France, Croatia, Hungary,
Italy, Portugal Portugal
Chlorpyrifos-methyl France, Italy, Croatia, 2-3 France, Italy, Croatia, Hungary
Hungary
Clorpyrifos-methyl-cypermethrin France, Hungary 1 France, Hungary
Cypermethrin France, Croatia, Italy, 2-3 France, Croatia, Italy, Portugal
Portugal
Deltamethrin France, Italy, Spain, 2-3 France, Italy, Croatia, Hungary,
Croatia, Portugal Spain, Portugal
Esfenvalerate France, Italy, Croatia 2-3 France, Italy, Croatia

Etofenprox France, Italy 1-3 France, Italy
Fenpyroximate Portugal 1 Portugal
Gamma-cyhalothrin France 3 France
Imidacloprid Portugal 2 Portugal
Indoxacarb Italy 1 Italy
Lambda-Cyhalothrin France, Italy, Hungary, 1-3 France, Italy, Hungary, Spain,
Spain, Portugal Portugal
Pyrethrum- Abamectin Italy Italy
Spinosyn Hungary Hungary

Spirotetramat Hungary Hungray
Tau-Fluvalinate France, Italy, Hungary 1-3 France, Italy, Hungary
Thiamethoxam France, Italy, Croatia, 1-3 Croatia, Portugal
Hungary, Portugal
Zeta-Cypermethrin France, Italy 1-3 France, Italy
Page 5
CONTROL OF FD VECTOR IN ORGANIC VINEYARDS
ACTIVE INGREDIENT USE AUTHORIZED IN USABLE IN NURSERY
Natural Pyrethrum France, Hungary In Hungary and Italy
Pyrethrum - Piperonyl Butoxide Italy Yes
Azadirachtin Italy, Portugal No
Pyrethrum-based products characteristics

Pyrethrum Pyrethrum
Name Dose/ha Regulatory status
content dose/ha
Use authorized in France and Italy against

Flavescence Dorée leafhopper
Use authorized in Hungary in emergency exemption

Pyrévert 20 g/l 1,5L/ha 30g/ha st th
between 1 July and 30 September on organic
vineyard and nurseries.
Maximum 3 application/year
Use authorized in Italy on grapevine against

PiretroNatura 40g/l 0,75 l/ha 30g/ha
Flavescence Dorée leafhopper
150-200
Biopiren plus 18,6 g/l Use authorized in Italy
mg/hl water
Pyrethrum Azadirachtin
Name Dose/ha Regulatory status
content dose/ha
Neemazal- T/S 10,6g/l 2-3l/ha 20-30 g/ha Use authorized in Italy one grapevine leafhoppers
In general, these active substances are more effective on juvenile forms of S.titanus. So, its use is
recommended when the insect population is constituted mainly of juvenile forms. Moreover, since the
persistence is not so high it is suggested to repeat treatment after one week. Products based on pyrethrum
are more effective along time if they are distributed in the evening or at early morning.
Conditions of pyrethrum application
Natural pyrethrum is a delicate molecule, sensitive to high temperature and UV radiation. Half-life time is
estimated to 10-12 minutes for a pyrethrum solution exposed to sun. Product has a “choc action” by contact
and act on insect nerve-conduction. In case of high infection by S.titanus of the plot, pyrethrum treatment
Page 6
need to be repeated one week after the first treatment. Application of natural pyrethrum can be done in
association with copper or sulphur application (Sudvinbio, 2013). Natural pyrethrum is effective on L1 to L3
stages of S.titanus. Unfortunately, efficiency of pyrethrum treatment is variable and serious monitoring of
vector population and observation of the plot need to be done regularly before and after treatment with
pyrethrum-based chemical specialities.
Recommendations for application
- Apply at the end of the day (low light and low temperatures)
- Water pH < 6.8 with an optimum between 6.0 and 6.5
- Apply quickly after mixture preparation
- Use a new can or one aged less than 6 months
- Maximum 3 application during vegetative season
Management of Flavescence Dorée in organic vineyards
Organic viticulture builds on prevention and on the implementation of a self-regulating system that
minimizes diseases and pest occurrence, increasing the functional biodiversity and, as a consequence,
minimizing the spaces for pest and diseases breakouts. For example, flora has an incidence on Scaphoideus
titanus populations, the vector can live both on grapevine and on flora between rows, with intensity of
population varying with time. Nymphs can develop, among other, on Trifolium repens and Ranunculus repens,
both common species used in vineyards (Trivellone et al, 2013). Moreover the presence of flowers
contributes to increase the population and diversity of several predators such as spiders that are founded
to reduce leafhopper population under cover cropped vineyards.
Biological control of S.titanus has been tried out either by releasing natural enemies from their native
habitat or by increasing local natural enemies’ populations but both strategies yielded poor results. Fitness
and efficacy of some biocontrol agents were assessed in relation to grape pest and insects. For instance,
the fungus Lecanicillium lecanii was used against S.titanus juvenile forms and proved to be virulent to the
second instar nymph stage.
Where prevention is not sufficient to keep FD under control, organic vine-growers in Italy use to spray
natural products allowed under EC Reg. 889/08, namely natural pyrethrum and Azadirachtin. As
explained above, they need to be applied with high precision and respecting certain specific requirements,
otherwise their efficacy (especially pyrethrum) is too limited.
A CARE FOR BEES AND POLLINATORS
It is important to remember to minimize the risk for pollinators while spraying insecticide on big territories
at the same time. Spraying must not take place at vine flowering time and they should not be localized on
flowers of vineyard’s inter-rows. Soil should be tilled or grass mowed before spraying. Moreover spraying
have to be performed in the evening after the sunset or in the early morning. In windy days spraying must
be avoided in order to reduce drift risk. In Italy the spraying period vary from year to another depending
to vine and vector phenological development. Generally the first treatment is performed after flowering
and at fruit set starting, to avoid damaging to bees.
Page 7
Be aware of Metcalfa or mealybug infestation, encouraging bees to visit the vineyard to pick up the
honeydew. In this case it’s better not to use a neonicotinoid for the second spray against adults; better to
use it eventually for the first treatment against nymphs, to control Metcalfa as well.
V I N E YA R D M A N A G E M E N T
1 - M O N I T O R I N G V I N E YA R D
Vineyard sanitation is a key point in FD management. Monitoring vineyard is one of the main key element
of FD management and must be done at scale of a territory and individual vineyard scale, often at
communal scale in order to have a collective implication in FD management. In order to be effective, the
better is that monitoring is organized and controlled by a dedicated organism but monitoring should be
done also individually by each winegrower on his own vineyard with respect of monitoring protocols.
Laboratory analysis can be done in case of doubt and is the only way to distinguish Flavescence Dorée
from Bois Noir.
2- INFECTED PLANTS MANAGEMENT

Uprooting or destruction of infected grapevine is ever suggested and mandatory in most of regions by
national decree. During period of leafhopper presence (from May to August) as soon as an infected vine is
detected it must be uprooted. The sooner infected vine is uprooted, the better is it, in this way Scaphoideus
titanus can’t feed on the infected vine, won’t become infectious and won’t participate to disease’s
propagation.
Uprooting need to be done rigorously avoiding any growth of suckers or rootstock. Rootstocks are healthy
carriers, they can host FD phytoplasma but not express any symptoms. Eradication of grapevine regrowth
need to be done in the vine-plot but also outside the plot. If a quick uprooting is not easy to do during the
growing season, it can be postponed after harvest, but the vine, or the symptomatic parts of the canopy,
even under suspicious, must be cut away as soon as possible.
Leaving infected grapevines in the plot will increase considerably infection level in the coming years.
DISEASE’S EVOLUTION
- Enlargement of disease area from grapevine to grapevine around initial grapevine

- Fast increase of diseased grapevines : amount of diseased grapevine can be multiplied by 10
each year (figure 3 and 4)
- After inoculation, delay in symptoms expression of one year.
Space-Time Point Pattern Analysis of Flavescence Dorée Epidemic in a Grapevine Field: Disease
Progression and Recovery
Page 8
3- WILD WINES MANAGEMENT
Wild vines host both the disease and the vector.

The various crosses of wild vines (Vitis rupestris, V. riparia, V. berlandieri etc.) and the ones between them
and European vines, used as rootstocks, are capable of hosting the FD phytoplasmas, although they
generally do not exhibit the typical symptoms of cultivated vine, or don’t show any symptoms at all:
therefore they are healthy carriers. The vector Scaphoideus titanus feeds indifferently on all species of the
genus Vitis, so it can acquire the phytoplasma and transmit the infection from wild to cultivated vines. In
addition, these areas can serve as refuges for the vector, therefore reducing the effectiveness of their
control.
THE FLIGHT OF SCAPHOIDEUS
The adult insect vector is able to make quite long flights and capable of dispersing from wild to cultivated
grapevine, (Alma, 2015; Lessio, 2014). However, the greater the distance of the "wild" outbreak from the
vineyard, the lower is the probability of a migration into the vineyard itself. The migration of insects from
outside towards inside a vineyard is also demonstrated by the greater frequency of the presence of
symptoms at the border of the vineyard. Most of insects move into the vineyard up to 20-30 meters from
hedgerows but in some cases it is possible that adults move up forward, wind allowing long distance
movments (Steffek et al, 2007)
HOW TO CLEAN UP?
It is not necessary to deforest. The vine is a sun-loving plant, looking for sunlight and then its canopy grows
mainly on the edge of forests, at the border of cultivated fields and roads. But if it is allowed to grow and
develop big canopies, it goes to ripen fruit, and at that point can also be propagated by seed, through the
birds. This should be avoided.
Cleaning in winter time Cleaning in springtime

After the harvest, since late autumn, until vine May-June: before treatment against the youth
budburst: cleaning of fallow stripes bordering the insects in the vineyard, destroying sprouts of
vineyard by paying particular attention to vines survived American vines is required. As an
climbing on trees. The debris must be removed and alternative, spray localized herbicides with a high
burnt, this step is very important because they can concentration of glyphosate* (from 2 to 5% solution
contain Scaphoideus eggs and could sprout from of the commercial product with 360g/L of
the cuttings and multiply. Where possible, eradicate glyphosate) in order to kill for starvation any young
the roots of the vine with an excavator. specimens of the vector.
*This protocol is not allowed in all winegrowing
regions and sometimes additional permits need to
be acquired due to potential environment pollution.
CAUTION: Do not destroy wild American vines in summer, because the adults may migrate from the
wild to the vineyard.
Page 9
The procedure must be repeated in subsequent years, trying to eliminate all American vinestocks.
Jan Feb Mar Apr May Jun Jul Aug Sept Oct Nov Dec
4 - I N N O VAT I V E A S P E C T S I N T H E M A N A G E M E N T
Alternative methods to chemical management of Flavescence Dorée vector are searched, mostly to be used
in organic production and in the objective to decrease pesticides use and protect the environment.
MATING DISRUPTION
Prevent Scaphoideus titanus population to grow from one year to another by disturbing its reproduction is
one innovative method in development. The method consists in disturbing signals emitted by males to attract
females and inhibit their reproduction. Adults communicate with vibrational signals, males have a “call and
fly” behaviour and emit a courtship song and females emits signals in responses to males (Mazzoni et al.,
2009). A mechanism broadcast vibrational signals through a supporting wire and the playback of
disruptive vibrational signals reduced the level of male calling and interrupted an established male-female
duet that consequently resulted in a significantly reduced number of copulations (Mazzoni et al, 2009).
PUSH PULL STRATEGY
“Push and pull” strategies involve manipulating insect behavior via the combined use of attractive and
repulsive items, including lures or plants capable of drawing pests into the area where they will be
destroyed. The use of this technique against vector of phytoplasma disease has already shown promising
results.
ORANGE OIL APPLICATION
Another alternative to chemical control of Scaphoideus titanus is the use of orange essential oil. This product
is used by some winegrowers in Europe to control S.titanus populations but complementary to chemical
treatment as essential oil is not registered as an insecticide. The active ingredient of orange oil is a
terpene, D-limonene and identified as a natural insecticide. The D-limonene as the properties to desiccate
nymphs and can be effective on the young stages of Scaphoideus titanus. Effectiveness of this product still
need to be demonstrated.
KAOLIN APPLICATION
The kaolin spraying has repellent function towards the leafhoppers; but some studies also prove mortality
of nymphs. It is mainly used in organic farming, where the only insecticide admitted and slightly effective is
the pyrethrum. It is not an alternative but a possible integration. Kaolin is more effective on early instars
than towards adults. Given the high cost of the product and it’s proven, even if partial, effectiveness, the
optimization of timing and doses should be further studied.
Page 10
BIOCONTROL AGENTS
Among other biological control agents, the use of endosymbionts, such as Wolbachia, seems to be
encouraging and also the symbiotic bacteria of genus Cardinium, which is responsible for impaired
reproduction and behaviour of the vector (Chuche et al, 2017). This bacteria specie was found in natural
populations of both S.titanus sexes with a high prevalence (94%) and in differents organs. Finally, S.titanus
also can host symbiotic bacteria of the genus Asaia, transmitted vertically by female as well by male
during mating, that can be at the same way during feeding. Some author have suggested using these
symbionts to decrease the vector’s capacity for S.titanus. Further studies should be performed.
5 - H O T WAT E R T R E AT M E N T : U S E I N N U R S E R I E S
France, Italy, Portugal, Croatia and Hungary have a national decree imposing mandatory treatments on
nurseries and according to the country they can be specific to some areas.
In some countries or regions there are specific rules regarding nurseries: mandatory hot water treatment, or
in some case prohibition to manage nursery activity in FD outbreak regions.
Hot-water treatment is proposed to cure dormant wood material from phytoplasmas and to supress surface
parasite and pests. Pathogenic agent, the phytoplasma is heat-sensitive. Time and temperature need to be
such as supress phytoplasma without affecting plant development capacity. The use of HWT should
intervene to complement preventive measures as insecticides treatments, suppression of contaminated
rootstocks and should supplement absence of chemical treatment against phytoplasmas. Propagating
material (either cuttings or grafted plants) are immerged into a bath of water at 50°C during 45 minutes.
These parameters allow to eliminate phytoplasma and to kill Scaphoideus titanus eggs (presents under
bark) (Caudwell et al, 1997).
Conclusion
A good knowledge on the 3 pillars involved in the FD fight, control of the vector, management of the
disease in vineyard and nurseries and regulation allow viticultural sector to be more efficient in FD
management. Being a complex diseases, all parameters are important, and a good knowledge of the
vector, its feeding behaviour and the evolution mechanisms of the diseases are essential to manage
precisely the intervention allowing to fight the disease. Furthermore, collaboration and commitment
between winegrowers, dedicated organisms, and other actors of FD management are indispensable in
effective disease management.
Page 11
Pictures
Picture 1: L1 nymph (IFV SOuth-West, IPTPO), L3 and L5 (INRA Bordeaux) (Fotó: Zsolnai Balázs /L1/, Varga
András /L2-L5/, Keresztes Balázs /imágó/)
Picture 2: Sticky trap (IFV South-West)
Picture 3 : Entomological umbrella (photo www.EntomolAlex-gr.com)
Picture 4: Scaphoideus titanus adult on a sticky trap (IFV South-West, IPTPO)

Picture 5: Wild vines (M. GIly, SIVE)
Picture 6: Mechanical system for mating disruption (Lucchi et al, 2013)
Picture 7: Kaolin application on leaves (ADVID)
Figures
Figure 2: Monitoring of S. titanus with yellow sticky traps in an area with several vineyard plots and wild Vitis
species – potential number and distribution of traps (white arrow: potential movement of S. titanus adult; red
dots: distribution of yellow sticky traps.
Figure 3: Insecticide treatment strategies. Adapted from « Jaunisses à phytoplasmes de la vigne, Groupe de
Travail National Flavescence Dorée, 2006 »
Fig.3: Flavescence Dorée spreading
Figure 4: Yearly maps of symptomatic Mt and newly symptomatic Mnt plants (left column), and dead Mj,
removed Mc and recovered Mr plants (right column). Maps are relative to years 2011-2015 during flavescene
dorée (FD) epidemic in the experimental site. Source : Federico Maggi, Domenico Bosco, Luciana Galetto,
Sabrina Palmano and Cristina Marzachì, 2016
Figure 5: Wild American vines management (D. Eberle, 2015)
REFERENCES
CONSULT WEBSITES:
http://pti.regione.sicilia.it/portal/page/portal/PIR_PORTALE/PIR_LaStrutturaRegionale/PIR_Assessorator
egionaledelleRisorseAgricoleeAlimentari/PIR_DipAgricoltura/PIR_AreeTematiche/PIR_ServizioFitosanitario
Regionale/PIR_Prodottidivulgativi/PIR_Allegati_Informazioneutenza/Link167_Cicaline.pdf
http://www.regione.piemonte.it/agri/area_tecnico_scientifica/settore_fitosanitario/vigilanza/dwd/15.pd
f
http://www.vignevin.com/fileadmin/users/ifv/publications/A_telecharger/JaunissesPhytoplasmesVigne.pd
f
http://hemiptera-databases.org/flow/?db=flow&page=home&lang=en
Page 12
APPENDIX 6: TRUNK RENEWAL
www.winetwork.eu
17/27
TRUNK RENEWAL
A practice applied in field to limit GTDs incidence

TRUNK RENEWAL
Introduction
Grapevine trunk diseases (GTD’s) are very old and present and anyway shows a very low vigour (Calzarano et al., 2004).
in vineyards since Antiquity. They cause damage all over the Results could be very much improved with the application of
world by attacking the vine stocks, going for more or less triazoles after the trunk renewal operation. Especially if the
long term, until the death of the vine. The symptoms are vi- renewal of the trunk has been carried out on a diseased vine
sible on the foliage, but also in the wood with the presence of and especially for the apoplectic forms. (Larignon P., Yobregat
several types of necroses. It is in the wood of the trunk that O., 2016.) (French National group of wood diseases, 2007)
the pathogenic fungi responsible for the GTD’s are concen- The renewal of the trunk only works if it is carried out early
trated. The technique of trunk renewal is consist to replace enough. Effectively on a vine expressing severe symptoms
the old trunk by a new one that is healthier. it is often too late. (SICAVAC, BIVC, 2015).
In nature, and for about 40 million years, Eurasian vines have However, many testimonials from winegrowers show that
been forest dwellers; vines have been multi-trunked and they are satisfied with the results. (Chamber of agricultu-
unpruned. It is probably in the past 5,000 years or so that re, Alsace, Burgundy, 2017) (DAL François, SICAVAC, San-
vineyards have been monocultures, with each vine trained cerre, 2017). Moreover, it can be seen that this practice is
to a single trunk. Multi-trunks are a practice used commer- very common in the regions of the Winetwork project, which
cially in places with severe winters to replace cold-damaged shows that it must provide some satisfaction.
trunks and it can be used to fight trunk diseases, too (Smart,
2015).
Description Application area

The trunk renewal technique is used to reform a new
trunk with a basal sucker. The advantage is to have
a new trunk, but with the well-developed root sys-
tem of a vine in production. Moreover, the renewed vine
will return to production faster than a young plant. The ef-
fectiveness of this technique is very good against
Eutypa (BNIC-INRA Bordeaux, 1989). The efficacy against
Esca and Botryosphaeria dieback is partial because
part of the renewed vines can express symptoms
again in the following years.
Scientific data
There is evidence that trunk renewal is effective in figh- Figure 1: Regions where trunk renewal is currently apply. Data from
interviews done during Winetwork project in 10 wine-growing
ting Eutypa by numerous scientific publications. The expe- regions
riments are sometimes old because Eutypa tends to decline
in most regions, unlike the Esca and BDA. (BNIC, Chamber Technique of trunk renewal is used in all regions of the Wi-
of Agriculture, 1989) (BNIC, INRA Bordeaux, 1989) Mette L. network project. It’s a really popular practice.
and al., 2004) (Sosnowski Mark R. and al., 2011) Several protocols are possible. Trunk renewal can be pre-
ventive, thus carried out before the appearance of symp-
On the contrary, there is less scientific data on the ef- toms of GTDs, or used when symptoms are observed.
fect of trunk renewal on the Esca and Botryosphaeria The two techniques can be complementary on the same plot.
diseases. The technique would seem less effective on The success of this method is related to the cultivar, disease,
these two diseases. Indeed, part of the vines whose trunk region and other parameters described later in this technical
was renewed express symptoms again in the following years data sheet.
Practical application
In the vineyard, there may be several scenarios were trunk in spring (figure 2).
renewal can be applied. The renewal of the trunk may be The important thing is to cut the trunk below GTD ne-
justified on a healthy vine that has risk factors for the de- crosis, to remove all the inoculum or at least (Calzarano et
velopment of GTD. We may have the case of a vine at the al., 2004) to apply effective fungicides after the treatment on
beginning of the disease that expresses the first symptoms the wound.
of GTD, or completely diseased vines. If trunk renewal is done in winter, the wound need to be pro-
tected to limit other infections into the wound (for example
There are also two types of trunk renewal, to be used ac- Eutypa lata fungi) (Larignon and Yobregat, 2016).
cording to the situation: “the classic renewal”, when a
vine shoot grows while keeping the whole vine in production,
or “the forced renewal” when the vine is decapitated to Explanation of the different steps:
force the development of suckers.
1. Make this technique on suitable grape varieties, or
1) Vines expressing GTDs symptoms: vines that produce enough suckers. (Look at the refe-
a forced trunk renewal rences at the end of the TDS)
It is possible to renew the trunk of a sick GTD vine and 2. Remove the symptomatic part of the trunk im-
therefore to replace an infected trunk with a healthy plementing a cut on the wood with a saw (or other ap-
wooden trunk. This helps improve yield and may slow the propriate tools like hydraulic vine shears). Protect the
spread of disease by removing potential inoculum sources pruning wound (with fungicide application, bio-control
from a vineyard. There is, however, no guarantee that agents…) and wait for the spring.
re-infection will not occur, and wounds made by cut 3. In spring, select a well-developed sucker from the
must be protected by fungicide application (Smart, 2015.), basal part of the trunk. Be careful; don’t take a develop
or bio-control agents. a sucker that belongs to the rootstock.
It is possible to decapitate the trunk before the harvest, as 4. Grow the sucker. If necessary, protect it against che-
soon as the first symptoms of GTD are observed. Indeed, the mical and mechanical weeding, for example with a plas-
technique seems more effective when the disease is tic grows tube.
not in apoplectic form but rather in slow form and at the 5. Disbudding of suckers on the stem, to stem training.
beginning of symptoms expression.
6. Prune the plant like a young vine. In order to form the
If the diseased vines have been identified, they can be cut desired training system.
during the winter. Different cultivars can react in a very diffe-
rent way to the winter cut as regards new shoots formation
Figure 2: Proceedings of trunk renewal (IFV Alsace).

TRUNK RENEWAL
Results : 2) Vines with no symptoms of GTDs but with factors

favoring the development of GTDs
This technique is very effective against Eutypa. For Esca
and BDA, this technique is more effective when the vines are This technique consists of reforming a new trunk be-
affected by the slow form. Indeed, for the vines affected by fore observing symptoms of GTDs. This is used to re-
apoplexy form, a part of new vines show symptoms juvenate the trunk to prevent GTD development.
again after 2 years. On the vine that were not apoplectic This can be done on a whole plot or on only vines depending
but with chronic form we observe a good recovery of the vine on several parameters.
but mortality after 4-5 years it possible. (Larignon P., Yobregat
O., 2016.) (French National group of wood diseases, 2007.) When a young plot (less than 15-20 years old) begins to
New trunks can be free of trunk disease infection if express symptoms of GTD, it is likely that the wood of a
located sufficiently low on the trunk, below wood majority of the vines is strongly necrotic and that the
necrosis due to the trunk pathogens. sap flux are altered. Invariably, the mortality rate will
increase in subsequent years. On these plots it is pos-
sible to restore the sap flux by renewing the trunks of all the
vines of the plot. To best renew the trunks, it is necessary to
choose a sucker as low as possible, to remove a maximum
of inoculum. (SICAVAC, BIVC, 2015).
In this situation, the old vine trunk is preserved and conti-

nues to produce with the double trunk method, it can
even serve as a tutor to attach the sucker (figure 3).
It is interesting to renew the trunks of a whole plot,
because the maintenance of vine leaves requires a speci-
fic manual work, almost similar to a young plantation. It is
easier to manage trunks renewal if it is done uniformly.
Suckers of 1 year old being lignificated after renewal of old trunk. One
selected sucker will form the new trunk (IFV South-West) It is also possible to renew preventively only a few vines, to
make timely trunk renewal when identifying vines with
Recommendation : problems that may favor GTDs, such as many or large size
of pruning wounds, lots of dead wood on the trunk, insuffi-
Renew as soon as first foliar symptoms appears. cient or disturbed sap flux.
Do not keep the old trunk as a stake, as necro-
sis are coming from the top of the trunk, where As in the previous case, the wound need to be protected
pruning wounds are located, and going down, new which will limit pathogens penetration into the wound.
trunk can be contaminated.
Wrong exemple of trunk renewal using the old affected trunk as a Trunk renewal in Istria, Croatia (IPTPO, K. Diklić)
stake. New trunk is affected now by GTDs (IFV South-West)
Figure 3 : Proceedings of preventive trunk renewal (IFV Alsace).
3) Timely trunk renewal Key points for succes

The technique was developped by Mr Richard Smart and The presence of suckers at the base of the trunk is a key
aims to retain health and recover yields before yield loss and point for success and depends on several factors:
manage GTDs in vineyard at an early stage of infection.
Protocol : - the cultivar: some cultivar are not suitable for trunk re-
newal because they have very low production rate of new
1-Assess GTDs infection by counting vines expressing foliar suckers on the base of the trunk. (Becker Arno, 2012)
symptoms (we can also have different level of assessment
accuracy, for visual rating to counting symptoms) and record (Examples of suitable cultivars: Colombard, Merlot,
also dead and missing vines. Timing of inspection depends Gewurztraminer, Auxerrois / no suitable: Ugni Blanc, Riesling,
on the predominant disease in the vineyard. For Eutypa it Portugieser…)
can be done in spring and for Esca and Botryosphaeriaceae - the mode of cultivation : remove suckers with chemi-
it can be done at end of summer, before harvest. cals is not favorable to the regrowth of suckers. In the same
2-Evaluate disease’s risk : according to cultivar there is diffe- way, a very rigorous remove suckers every year is not favo-
rent susceptibility to GTDs rable to the development of buds from the old wood.
3-Combining risk and infection to decide management - the age of the vines: the younger vine produce more
strategy. suckers. With the increasing age of the grapevine you have
less and less sleeping buds around the grafting point. You
According to the risk of the plot (cultivar susceptibility, global should do the trunk renewal not later than the age of 20
management) and the present infection rate of the plot 4 years of the vine to get positive results. (Becker Arno,2016)
management strategies were proposed by Richard Smart:
• S1 (low risk, low infection): remove dead vines, keep suc-

kers in the vine presenting the symptoms of GTDs, and re-
new them.
• S2 (low risk, medium infection; medium risk low infection):
pre-harvest inspection to identify early stage of symptomatic
vines, remove dead vines, trunk renew symptomatic vines
and adjacent vines if clumping and staining are evident
• S3 (low risk, high infection; medium risk, medium and high
infection; high risk, low and medium infection): same as S2
but all trunks of the plot will be replaced within one or two
years.
• S4 (high risk, high infection): for all vines Winter-prune very
hard, remove all trunks in spring and replace all trunks (or
replant) Renewed trunk in Eger, Hungary (Eszterhazy Karoly Unversity)
TRUNK RENEWAL
Help to bud burst the eyes of the base of the trunk : Practice cost
If buds are present at the base of the trunks, it is possible to GTDs cause significant economic damage. They can reduce
help them to budburst by eliminating the excess thickness the yield and quality of the grapes. This reduces the quantity
of dead bark and by making a superficial notch with the and quality of the wines produced and therefore the pro-
pruning shears just above the zone where a departure of fitability of the vineyard. For table grapes, symptoms with
sucker is desired (SICAVAC, BIVC, 2015) black punctures on the berries can make the production un-
marketable. Some of the economic losses can be offset by
techniques that limit damage.
Method : remove the old bark from the trunk and make a
notch two millimeters deep on 3-4 centimeters long at the Overall, the implementation of preventive practices
base of the trunk (figures 4 and 5). to fight against GTD’s is profitable. Indeed, using pre-
Other key point: Good knowledge of GTDs symptomatology ventive practices early makes it possible to lengthen the pe-
about foliar symptoms and internal symptoms. When cut is riod of profitability of the vineyard. In some cases it increases
done, it is important to know to identify necrosis, and if there profitability immediately. In any case, using preventive prac-
is one, to cut below. tices, before GTD’s appear is positive economically. (Kaplan
J. and al., 2016)
The cost of remediating vines by working or re-grafting will be
less than the cost of replacing vines. The sooner preventive
practices begin, the greater the future benefit. (Sosnowski M.
and Mundy D., 2016)
According to Becker (2012) profitability of the trunk renewal

depends on the expected price of the wine and on the remai-
ning useful life of the vineyard. The trunk renewal is eco-
nomically justified if the reaming useful life time of
the vineyard is ten years, wine price of 0,8 € per liter
and a success rate of the trunk renewal of 20%. When trunk
renewal is applied with 60% of success (symptomatic vines
were ‘converted’ into asymptomatic vines), the cost of trunk
renewal resulted in 3 € per vine. Replacement of a sympto-
Figure 4: Remove bark (SICAVAC- BIVC) matic vine by a young plant costs 9 €/vine.
Concrete example:
Quantification of a trunk renewal cost according to a sur-

vey of winegrowers in the South West of France, in the
Winetwork project:
In the case of a plot planted with 4500 vine / ha, and an
average of 250 vines to be renewed :
• Practice cost : personal cost around 15€/hour (gross
cost), around 10 hours of work/ha (to realize the trunk
renewal)
• Stake: 0.10€ in wood (bamboo) and 0.30€ in iron,
plastic protection: 0.20€
Total cost: 225€/ha/year maximum (with bamboo
stake) to 275 €/ha (with iron stake).
Figure 5: Making a notch (SICAVAC- BIVC)
Becker Arno, DLR Rheinhessen-Nahe-Hunrück, Oppenheim, Ger- Larignon P., Yobregat O., 2016. «Cahier pratique: comment lutter
many, 2016. «Alte Reben – Neues Leben», «Landwirtschaftliches contre les maladies du bois de la vigne?» «Practical notebook :
Wochenblatt LW 7/2016», page 38 to 41. how to fight against the diseases of the wood of the vine» IFV,
page 7.
Becker Arno, DLR Rheinhessen-Nahe-Hunrück, Oppenheim, Ger-
many, 2012. «Esca-Sanierung durch Stammrücknahme lohnt», Mette L., Creaser, Wicks Trevor J., 2004, «Short-term effects of
«Der Deutsche Weinbau, Nr.3», page 22 to 24. remedial surgery to restore productivity to Eutypa lata infected
vines», Phytopathologia Mediterranea, 2004, 43, 105-107.
BNIC-Chamber of agriculture, 1989. « Le recépage, ça marche ! »
« The trunk renewal, it works ! », experimental results. Richard Smart, 2015. Timely trunk renewal to overcome trunk di-
seases, practical winery and vineyard, oct 2015, p 64-70.
BNIC-INRA Bordeaux, 1989. « La pratique du recépage dans la
lutte contre l’eutypiose » « The practice of trunk renewal in the fight Sosnowski M. and Munday D., 2016. Sustaining vineyards through
against eutypa », Le paysan Français, n°861 november 1989. practical management of grapevine trunk diseases, NZ Winegrower,
August/september 2016, page 85 to 88.
Calzarano F., Di Marco S., Cesari A., 2004. Benefit of fungicide
treatment after trunk renewal of vines with different types of esca Sosnowski M., Wicks Trevor J. and Scott Eileen S., 2011, «Control
necrosis. Phytopathologia Mediterranea, 43(1), 116-124. of Eutypa dieback in grapevines using remedial surgery», Phyto-
pathologia Mediterranea, 2011, 50 (supplement), S277-S284.
Groupe National Maladies du bois ; French National group of wood
diseases (IFV, INRA, ENITAB, Minister of Agriculture, Viniflhor), SICAVAC, BIVC, 2015. «Manuel des pratiques viticoles contre les
2007. «Note nationale: les maladies du bois» ; «National note : maladies du bois», «Handbook of wine-growing practices against
Diseases of wood » . page 5. wood diseases», page 85 to 90.
Kaplan J, Travadon R., Coper M., Hillis V., Lubell M., Baumgartner
K., 2016. “Identifying economic hurdles to early adoption of pre-
ventative practices : the case of trunk diseases in California wine-
grape vineyards.”

Thanks to Prof. Laura Mugnai and
Dr. Vincenzo Mondello who partici-
pated to the development of this
datasheet.
APPENDIX 7: INTRODUCTION OF SMALL WOOD STICKS
INOCULATED WITH TRICHODERMA
www.winetwork.eu
18/27
Insertion of wood dowels inoculated with

Trichoderma sp inside grapevine trunk
It shows 2 samples of this type of practice in two wine Designations of Origin in Galicia
(Spain): Ribeiro and Valdeorras. The practice consists of inserting Trichoderma
inoculated wood dowels, into small drilled holes in the base of a trunk in order to fight
against Esca and Eutipiosis diseases.
T R I C H O D E R M A I N O C U L AT E D I N D O W E L S T O
I M P L A N T I N T RU N K O F V I N E
GENERAL PRINCIPLE
The technique consists of inserting into the trunk of a vine, small wood dowels inoculated with Trichoderma sp,
by drilling small holes into the trunk. Two examples of this practice in two wine Designations of Origin in Galicia
(Spain): Ribeiro and Valdeorras are described in this datasheet.
BACKGROUND
The dowels were manufactured by New Zealand company which has carried out research on on this kind of
treatment for a decade.
Instructions based on essays in both field and laboratory for using this product, indicate that these dowels are
working against Eutypa Lata and some species of Botryosphaeria. It is a preventive treatment, so it is
recommended to carry it out as soon as possible in healthy vines or in those where the affected part is
eliminated and the healthy part is treated.
In Galicia, there were two winegrowers applying this practice. Both cases seemed to have plants affected by
Esca and also Eutipiosis. Both of them used dowels that came from NZ commercial house
However, it must be taken into account that conditions of application of this treatment were not implemented
in accordance with the recommendations for this type of treatment. The cases which are showed below
occurred in very adult vineyards. Symptoms seemed to respond more to Esca and Eutipiosis and plants had also
showed symptoms for several years (8-10) before carrying out this practice.
A P P L I C AT I O N C A S E S
1ST CASE. APPLICATION IN A VINE AGED OVER 100 OF VARIETY DOÑA BLANCA WITHIN DENOMINATION OF
ORIGIN RIBEIRO
This is a particular vine of huge dimensions and high yield (50-70 kg per year), which does not seem to have
been grafted but planted directly.
This vine started showing the first symptoms in 2006 and these symptoms were a mix of Esca and Eutipiosis
(necrosis formed around the pruned arms, shoots with short knots, smaller leaves and sometimes with
necrosis on the edges plus absence of blooms and clusters). The rotten wood had a brownish or salmon color.
Page 1
Ilustration 1. Appearance of autochthonous cultivar Doña Blanca in 2006
Application area:
The application area is located in the South of Galicia, parish of Lebosende, Leiro within Denomination of Origin
Ribeiro. This area is characterized by an annual average temperature of 13,6 ºC and 17,8 ºC between April and
September , with a total rainfall of 1.182,7 mm and 220.5 mm during the vine’s growing period.
Specific application:
The application took place on the 26th April 2016. The phenological stage of grapevine was BBCH 11-13=1-3
leaves unfolded. The sap was flowing completely.
Before the drilling took place, dead bark was removed until healthy timber was reached. 3 wood dowels
inoculated with Trichoderma were inserted, 1 in the base of trunk below sucker which arised in 2015 and the
other 2 in the main arms.
Days after the insertion the rotten wood on each arm was cleaned out.
Ilustration 2. Three drilled holes. One in base of

trunk and two in the two main arms
Page 2
2º.CASE. APPLICATION IN A VINEYARD OF 0.5 HAS OF GODELLO CULTIVAR
Cultivar: Godello
Rootstock: Richer 110
Vine age: 29 years
Planting Density: 2,25x 1,2 m
Training system: double cordon Royat with 3 spur each side
Pruning system: Spur pruning (2 buds/spur and 12 buds/vine)
Yield: 8000 kg/Ha
Pruning debris:
Soil management:
Application area:
Area in SE Galicia within Designation of Origin Valdeorras. This area is characterized by an annual average
temperature of 13,9 ºC and a temperature of 18,8 ºC between April and September. A total rainfall of 1.118
mm and 274 mm during the vine’s growing period.
Soil : Loamy, deep, fertility medium-high, fresh. PH: 6.5
Symptoms of Esca appeared 8 or 10 years ago, starting with Chlorotic leaves , necrosis and dried canes and
shoots and ended up with the death of the plant.
Specific application:
In order to be able to adapt the treatment to time
availability, the plot of land was divided into 4
blocks, applying the treatment to each of them in
consecutive years. Treatment was applied into the
1st and 4th block in 2013- in October and April
respectively. It was applied into the 2nd block in April
2015 and into the 3rd block in May 2016.
Symptomatic plants were marked in summer in
each of those years, rating the severity of affection
from 1 to 5 as follows:
<25% : 1 point
Ilustration 3. Treated plot of Godello cultivar 25-50%: 1 point
50-75%: 2 points
75-<100: 3 points
=100: 5 points
Implantation in affected plants was made treating 3 out of 4 of them, leaving one for control. 2 wood dowels
were inserted into each vine in both sides of the base of trunk with differences of 1-2 cm of height.
Dowels used in 2013-2015 were from producer Vinemax (Agrimm). The same as the ones used in the first case.
The ones used in 2016 did not belong to the same producer although they were the same strain as the ones
used for the other treatments.
Implantation of Trichoderma dowels was made on the 26th May 2016, with vines in phenological BBCH
53=inflorescence visible
Page 3
Ilustration 4.- Test design
Ilustration 5. Symptomatic leaves
Ilustration 6. Drilling in Godello cultivar
SOME SCIENTIFIC ELEMENTS
It is no clear which is the mechanism of control that can reduce the incidence on treated vines. It is considered
that the main mechanism is not the bio-inoculant growing throughout the plant but rather the elicitor
response on the part of the vines. “Trichoderma has been shown to be capable of eliciting a systemic acquired
resistance (SAR) response in plants, which can raise the levels of resistance to diseases by stimulating
phytoalexins, which are natural plant defense chemicals ".
The researcher John S Hunt showed that Trichoderma can live in healthy grapevine tissues in association with
the pith parenchyma cells. F u r t h e r m o r e, other experiments performed by John have shown that a
particular T. harzianum strain, when inoculated into healthy vines, actively grows through the tissue over time.
It has been isolated up to 18cm from the site of inoculation 18 months later(6). These observations suggest
Trichoderma has the potential to be applied as a treatment to vines for a prolonged protective effect.
Page 4
OUTCOMES
It must be considered that treatment applied in very old vines (as it is presented in both cases, especially the
first), have low effectiveness.
To obtain statistically significant data, it is necessary to carry out the treatment in a high number of vines and
for more than 3 years since the symptomatology of this type of disease is erratic (some years vine is expressing
symptoms and others do not)
1º. CASE. APPLICATION IN AUTOCHTHONOUS VINE, DONA BLANCA OVER 100 YEARS
The same year of implantation, 2016, the vine showed symptoms of recovery, with shoots and fruit set in parts
that were affected.
Although the production obtained, approximately 3 kg, was very low, it was much greater than in the previous
year in which the vine produced practically nothing. This can be significant if we also take into account that that
summer there was a heat stroke, with high temperatures causing severe stress on vines.
The same treatment was carried out by the same winegrower in 2012 in 5 vines that showed the mainly
common symptoms of Esca. 3 were of Albariño cultivar and 2 were of Treixadura. One of the latest suffering
from severe attacks.
Nowadays, after 4 years, the 3 vines of Albariño have recovered and are giving normal productions. Regarding
the Treixadura, 1 is working regularly and the other is dead.
However, the number of treated vines is very low, so the results are not representative.
2º.CASE. APPLICATION IN A VINEYARD OF 0,5 HAS OF AUTOCHTHONOUS VINE GODELLO
There is plenty of data from the assessment of damages that has been taken on each block for 3 years, which
should be studied statistically. However, in a first general assessment it can be observed that:
1º.-In all the blocks, the number of affected vines has increased since first treatment. New affected vines appear
apart from those already registered.
2º.- Comparing registered assessment for 3 years, the incidence of damage on treated plants (sum of affected
plants) is much lower. But it occurs not only to treated vines but also to control plants. Therefore, it could be
said that the treatment is not having effect.
However, it has only been analyzed for 3 years and this type of diseases requires longer time for analyzing. So
registered data of plants treated last year and 2 years before should be analyzed in more detail, as well as the
treatment conditions (insertion was not always done in the vegetative rest of vine, level of affection of the
plant, age,…)
KEY POINTS FOR SUCCESS/ RISKS

1º.-Currently this kind of dowels seem to have effect against Eutypa Lata and Boryosphaeria stevensii
2º.- As it is considered a preventive treatment, it is more advisable to carry it out before the plant gets infected
- in order to be able to resist the attack of fungus.
Page 5
In case of healthy vines, it is advisable to drill a hole at a slight upwards angle 28 mm deep x 6 mm
diameters into the trunk below the cordon and insert one Bio-implant.
For a moderate incidence level which only affects the cordon: Cutting-off affected cordon wood,
then drilling into clean trunk wood below the cordon and make the insertion of one bio dowel per
vine.
For a high incidence which has reached the trunk it is recommended to replace by a suitable
sucker at base of trunk and drill it as close as possible to ground level or 25 to 50 mm below the
position of the sucker and to apply 1 Bio-implant per vine.
3º.- Indications of the commercial company state that it should be treated when sap does not flow. Avoid late
winter or early spring, when the sap starts flowing.
4º.- It ensures a protection for 4 to6 years. Then treatment should be repeated.
5º.- Since these implants contain living organisms, it is indicated to ensure there is a 24 hours period between
its applications and the use of any fungicidal spray applications.
I N N O VAT I V E A S P E C T S
This form of application has been tested in other fruit species besides vineyards for the control of other
diseases. Nevertheless the way to apply the Trichoderma by insertion of dowels is not usual and for what is has
been observed in essays, it seems to protect the vines for several years. It has the difficulty of the time spent
for its application and therefore its greater cost.
OTHER REQUIREMENTS
The time required for this application is about 2 minutes/ plant
Warning: In Spain Trichoderma sp is not registered and authorized for application against GTDs to grapevine, and
therefore this type of implants are not allowed either.
SOURCES OF INFORMATION
John S Hunt . (2004). Trichoderma and trunk disease fungi: prospects for new protective management
options. 2004. The Australian & New Zealand Grapegrower & Winemaker.
Mike Roberts (2012). Vinevax Bio-inoculant trichoderma treatment for Eutypa shows promise in Henschke
Vineyards. Grapegrower and Winemaker Issue 582, 2012
John, S., Scott, E.S., Wicks, T.J. and Hunt, J.S. (2004). Interactions between Eutypa lata and Trichoderma
harzianum. Phytopathologia Mediterranea 43, 95–104.
Mark Sosnowski. Dr Richard Lardner . Research Organisation: Cooperative Research Centre for viticulture.
July 2006. Diagnosis and management of eutypa dieback. FINAL REPORT TO GRAPE AND WINE RESEARCH &
DEVELOPMENT CORPORATION. Chief Investigator: Assoc Prof Eileen Scott.
http://winetitles.com.au/gwm/view/?action=view&id=930
MORE INFORMATION
FA of Winetwork project
Email: mjfanjul@xunta.es
Tel: +34 881997277
Page 6
APPENDIX 8: PRUNING WITH REGARD TO SAP FLUX
www.winetwork.eu
19/27
Pruning with regard to sap flux
Management of grapevine trunk diseases (GTDs) is oriented to preventive control strategies that reduce new infections and
disease spread. Winegrowers are applying new strategies in pruning with a belief that GTDs incidence and severity would be
reduced. Even if more detailed scientific results are still missing, this technical datasheet aims to present the current findings
and experiences in implementing such approach.

Description
1- Traditionnal pruning fection pathways of GTD fungi (Úrbez-Torres and Gubler,
2010). To reduce the risk of infections with fungi through
Traditional training systems and pruning methods pruning wounds, training systems that minimize the number,
had been primarily oriented to achieve adequate yield and size and accumulation of pruning wounds on grapevine pe-
good quality fruit, but impact of training systems on rennial trunk are preferred (Surico et al., 2008) (Fig. 1 - 2).
diseases, such as fungal trunk diseases, was neglected.
Factors related to pruning such as training system, weather Traditional training systems with numerous large and raze
condition during pruning period, number and size of pruning pruning wounds on the grapevine trunk potentially interrup-
wounds, location and accumulation of pruning wounds, cane ted grapevine sap flux and reduced grapevine longevity. If we
and spur length, wound protection, wound age, period of take into consideration that the pruning potentially induces
pruning, pruning debris management potentially contribute a 1.5 times larger necrosis in comparison to pruning wound
to the risk of infections with grapevine trunk diseases fungi size (Crespy, 2006), the trunk of the vine presented has a
(GTD). Pruning wounds are one of the fundamental in- significantly reduced sap flux.
Figure 1: Increased number of small and large wounds accumulated on upper part of grapevine trunk of cv Muscat blanc and Istrian Malvasia (in-
dicated by circles). Training system is Guyot-double. (K. Diklić, IPTPO)
Figure 2: Fig. 2: “Return cut” made in old vineyard to replace the cordon training system with Guyot-double training system (A, B) or to lower the
cordon (C) (indicated by circles and arrows), (K. Diklić, IPTPO).
2- Innovative/alternative pruning
Pruning in regard with sap routes (Guyot-Poussard,

‘modified’ spur-pruned cordon and other training systems) is
implemented in a more significant scale recently in European
winegrowing regions and many winegrowers expect to have
significant results with this practice in the upcoming years
(Fig. 3 - 5). The impact of this pruning approach on
grapevine trunk diseases, in comparison with traditional
training systems, still needs to be scientifically eva-
luated.
Currently, a diffused hypothesis is that pruning in regard
with sap routes reduces the probability for new infections
thanks to the small size and low number of pruning wounds.
Some training systems require retraining and a return cut,
common in older vineyards, which could be avoided with
this method of pruning. Moreover, large and raze wounds on
perennial wood, common on retrained and old vines found
in traditional vineyards, appear to be more sensitive to GTD
fungi infection than wounds on 1-year-old wood (Moller and
Kasimatis, 1980).
Figure 4: Pruning in regard with sap routes implemented in Istria

region on cv Teran, pruning made by Coronica winery. (up: prior
pruning, down: pruned vine). (K. Diklić, IPTPO)
Figure 5: Pruning in regard with sap routes implemented in Istria

region on cv Teran, pruning made by Coronica winery
Figure 3: Pruning in regard with sap routes, pruning made in the area
of north Italy. (up: Guyot-Poussard, down: ‘modified’ spur-pruned
cordon). (K. Diklić, IPTPO)
Technical and scientific data

The cut diameter of pruning wounds induces a 1.5
times longer necrosis on pruned spur or cane (Fig. 6, b)
(Crespy, 2006). Large wounds and raze cuts near perennial
parts of grapevine cordon and/or trunk induce wood necrosis
(Fig. 6, b), that potentially leads to higher infection rates
with GTD fungi (Úrbez-Torres and Gubler, 2010) and dete-
rioration of sap flux (Crespy, 2006). Moreover, there is
a hypothesis that deterioration of grapevine sap flux would
increase the negative impact of GTD due to higher stress in
grapevine physiology (Simonit & Sirch, 2010). Development Figure 6: Correlation between pruning wound and necrosis (cone of
of necrotic wood tissue may reduce water transport through desiccation) development (left: one year-old spur and cane; right: raze
xylem to leaves (Maher et al., 2012), and high transpiration wounds near perennial wood) (Crespy, 2006).
demand may lead to apoplectic forms (Surico et al., 2005).
It is important to prune correctly in order to minimize the

possibility of new infections with GTD fungi (Fig. 7, a) and
preserve a functional grapevine sap flux forming a comple-
mentary cone of desiccation (Fig. 7, b). Pruning techniques
that prioritise a “clean cut” (Fig. 7 c) are aesthetic and most
of the time lead to deterioration of grapevine sap flux and
formation of large wounds.
Pruning in regard with sap flux was adopted by Lafon (1927),

Figure 7: Recommended method of pruning (Crespy, 2006).
from a training system used in France, and later named
Guyot-Poussard according to its developer (Lecomte et
al., 2011). The main principle and difference of the traditio-
nal training systems (Fig. 8) and this training system is the
maintenance of the same sap flux from one year to another
with pruning that positions wounds only on the upper part of
the cordon (Fig. 9).
At our knowledge, pruning in regard with sap routes is imple- Figure 8: Traditional pruning: pruning wounds located on perennial
mented in training systems Guyot-Poussard and modified wood causing deterioration of grapevine sap flux (Simonit & Sirch).
spur-pruned cordon, but it is in development process
for training systems Gobelet and Pergola (Simonit & Sir-
ch, SICAVAC). Pruning in regard with sap routes has been
developed in some winegrowing areas in Europe, where
Guyot-Poussard is the most frequent training system that
respects sap flux but further implementation of this approach
in other training systems may be expected.
More detailed scientific research that will provide precise

answers on the efficiency of these training systems in GTD
preventive management are in progress and the impact
Figure 9: Guyot-Poussard: pruning oriented to preservation of grape-
of these training systems on GTD is not yet fully un- vine sap flux. (Abbreviations: black triangles - pruning wounds, red
derstood and needs to be scientifically evaluated! line – sap flux). (http://simonitesirch.com)
Guyot-Poussard training system
Application area
At the end of the first year after vineyard establishment a
Pruning in regard with sap routes is an innovative practice la-
fruiting cane, which follows the sap flux, is left and cut on a
tely implemented in winegrowing regions of Europe (Fig. 11).
2-bud spur. In the second year one developed fruiting cane
is left and cut on the height of the first wire (Fig. 10, a - d).
Formation of the central part of the vine – fruiting canes are

formed in a central part, one main fruiting cane is selected,
bended on the first wire while other are pruned on 2-bud
spurs (Fig. 10, e – h). In comparison with standard Guyot,
shoots are developed at the same horizontal level from the
1-year-old cane. “Ramification” is formed and horizontal bi-
lateral growth is achieved with perennial spurs that ensure
continuous sap flux (Fig. 10, i).
In the upcoming years, pruning cuts are always made
on the upper part of the perennial trunk. The pruning
technique that preserves a cone of desiccation that does not
alter the sap flux and fruiting cane developed on terminal
parts of the perennial structure ensure the longevity of this
Figure 11: Area of application Guyot-Poussard pruning (signaled by
training system (Fig. 10, l - m). red dot), Results from WINETWORK interviews.
Figure 10: Guyot-Poussard formation (Simonit & Sirch, 2010)

Innovative aspects Other requierements and costs

A lot has been done, but a lot more to do! The change from the traditional training system to the
pruning in regard with sap routes was conducted by Pet-
First partial scientific results from Ger- gen (2016 a, b) at the DLR Rheinpfalz in a seven-year-old
many – work in progress! vineyard with the grapevine variety Riesling. A higher amount
of work in the first years of the training system retraining
First results obtained from trials conducted in Germany (Pet- to Guyot-Poussard was observed. For the Guyot-Poussard
gen, 2016 a, b) indicate significant differences in the cut pruning there is a need of 37 h/ha whereas for the traditio-
surfaces between traditional pruning and pruning in regard nal pruning only 23 h/ha working time. These differences
with sap routes. Pruning in regard with sap routes results occurred especially because of the lack of long ex-
in increased cut surfaces in the annual wood, but the cut perience in adopting this training system, in comparison
surfaces of the perennial wood on central upper part of the with traditional pruning method. In the following years, after
trunk, common in Guyot pruning (see Fig. 13) were smaller a successful retraining, time saving is observed in the
than in the traditional pruning. future pruning.
Winegrower’s experience with pruning in The starting situation of a traditionally pruned vineyard plays
regard with sap flux an important role of the length and success of the change
to the Guyot-Poussard training system. Therefore the age,
Pruning in regard with sap routes, more precisely Guyot-Pous- variety and the growth of the grapevines plays a significant
sard training system, ensures a continuous sap flux and role. When there are different starting situations of retraining
horizontal growth along the first wire as a result of conti- to Guyot-Poussard, each situation requires an individual ap-
nuous horizontal development of adjacent perennial spurs. proach. There were no significant differences in the Botry-
In comparison with traditional Guyot pruning, where pruning tis bunch rot susceptibility and analysis of the grape-must
wounds are accumulated in the complete central-upper part between both pruning methods.
of the trunk (Fig. 1), Guyot-Poussard maintains the sap flux The study indicates that the change from a traditional-
because of the location of pruning wounds on upper part ly pruned vineyard to the Guyot-Poussard training
of the perennial wood (Fig. 13). This training system is im- system take several years. Guyot-Poussard is a de-
plemented in the production of high quality wines, and it is manding pruning method and has to be learned before
believed that contributes to more homogenate development and temporary workers need to be properly trained.
of phenological stages, balanced vegetative growth, more Because of the long incubation time of the Esca pathogens
balanced ripening (significant for production of red grapes at least ten years for the first observations and results on the
designated for structured red wines). Activities such as shoot impact of this training system are required.
removal are more extensive and important to maintain the
training system, but leaf removal at phenological stage of
flowering or fruit setting requires less man power.
More information
Technical data sheet «good pruning practices»
KNOWLEDGE RESERVOIR: www.winetwork-data.eu
Pruning in regard with sap routes implemented in Istria region on cv

Teran (K. Diklić, IPTPO)
Crespy, A. (2006). Manuel pratique de taille de la vigne. (Ed. Simonit & Sirch. http://www.aloislageder.eu/sites/default/
Oenoplurimedia). files/interview_marco_simonit_1.pdf
Geoffrion, R., Renaudin, I. (2002). Anti-esca pruning. A use- Simonit, M. (2014): Manuale di potatura della vite Guyot. Ed.
ful measure against outbreaks of this old grapevine disease. LÌnformatore Agrario, Verona.
Phytoma. La Défense des Végétaux (France). Simonit, M. (2016). Cordone speronato. Ed. L’Informatore
Lafon, R. (1927). Modifications à apporter à la taille de la Agrario, Verona.
vigne dans les Charentes. Taille Guyot-Poussard mixte et Simonit, M., Sirch, P. (2010). Il metodo Simonit&Sirch, Pre-
double. L’apoplexie, tratement préventif (Méthode Pous- paratori d’uva. Potatura ramificata per la logevita dei vigneti:
sard). Traitement curatif. Imp. Roumégous et Dahan, Monte- Osservazioni teoriche e guida pratica per Guyot e cordone
pellier, 1921. speronato. http://www.vitevinoqualita.it/files/2013/07/pota-
Maher, N., Piot, J., Bastein, S., Vallance, J., Rez, P., Gué- turaramificata_it.pdf
rin-Dubrava, L. (2012). Wood necrosis in Esca-affected Surico, G., Mugnai, L., Marchi, G. (2005). Older and more
vines: types, relationships and possible links with foliar recent observations on Esca: A critical overview. Phytopa-
symptom expression. J. Int. Sci. Vigne Vin., 46 (1), 15-27. thologia Mediterranea, 44, S68-S86.
Moller, W.J., Kasimatis, A.N. (1980). Protection of grapevine Surico, G., Mugnai, L., Marchi, G. (2008). The Esca disease
pruning wounds from Eutypa dieback. Plant Disease 64, complex. In: Integrated management of diseases caused by
278–280. fungi, phytoplasma and bacteria. (Ciancio A., Mukerji K.G.,
Petgen, M. (2016a). Sanfter Rebschnitt in der Umstellungs- eds.). Springer Science+Business Media B.V., 119-136.
phase. Das Deutsche Weinmagazin. Úrbez-Torres, J.R., Gubler, W.D. (2010). Susceptibility of
Petgen, M. (2016b). Erste Erfahrungen nach der Umstellung. grapevine pruning wounds to infection by Lasiodiplodia
Das Deutsche Weinmagazin. theobromae and Neofusicoccum parvum. Plant pathology,
SICAVAC http://www.vinopole.com/fileadmin/user_upload/ 60(2), 261 – 270.
fichiers_vinopole/Maladies_du_bois/Formation_taille_
guyot_resume.pdf
SICAVAC, BIVC (2014). Manuel des pratiques viticoles contre
les maladies du bois Remise à jour du «Guide pratique de la
taille Guyot»

Thanks to Dr Philippe Larignon and
Dr Vincenzo Mondello who parti-
cipated to the development of this
datasheet.
APPENDIX 9: TRUNK CLEANING
www.winetwork.eu
20/27
TRUNK CLEANING
A practice applied in field in to limit GTDs symptoms expression

TRUNK CLEANING
and show no efficacy for Eutypa (Thibault M, 2015, La-

Introduction on the principle rignon P., 2016). Trunk cleaning can be applied every time
Cleaning grapevine trunk is a technique developed to limit of the year.
the grapevine trunk diseases (GTDs). Grapevine trunk di- However, winter period is more convenient. In this case, di-
seases are an important cause of European vineyard decline, seased vine need to be identified and marked during
in the medium to long term GTDs lead to the death of grape- summer (September is the best year period to see the
vine. Grapevine trunk diseases are caused by fungi dama- symptoms). If trunk cleaning is made in summer harvest can
ging grapevine functional wood. be saved and the grapevine can lignified normally. Disadvan-
Trunk cleaning can be applied only on Esca and Botryos- tage at this period is the lack of time.
phaeria dieback, furthermore, more superficial are The technique can be applied on different grape varieties
the symptoms, more efficient is the technique. (such as Pinot noir, Sauvignon blanc, Chardonnay, Cabernet
The objective of the technique is to suppress white rot, franc and Cabernet sauvignon) and in different climates, al-
called «amadou» avoinding cutting functional sap flux. White titudes and terroirs.
rot need to be take-off entirely otherwise grapevine could
express again GTDs symptoms. White rot is the result of suc- Application area
cessive action of several fungi: Phaeomoniella chlamydos-
pora, Phaeocremonium minimum, Eutypa lata, Fomitiporia Trunk cleaning is an innovative practice applied in few re-
mediterranea that will deteriore the dead wood created by gions. In France and Italy winegrowers are doing it themsel-
the previous fungus. ves and sometimes are calling on specialised company who
realized the cleaning.
The trunk cleaning technique is a modern tree surgery,
also called «dendrosurgery» based on a 100-year-old tech-
nique to heal plants affected by the Esca disease, and to
remove the infected wood. This technique is, thus, not a new
one, but rather the recovery of an ancient practice known
since ancient times, as Ravaz and Lafon (1927) described,
and put into practice with new tools and knowledge. Pous-
sard employed it at the end of the 1800s with encouraging
results: 90-95% of success rate.
Trunk cleaning is a “surgical” technique experimented firstly

in France and then in Italy, which entails the removal of
the decay that forms beneath wounds caused by incorrect Figure 1: Areas where trunk cleaning is applied. Result from
pruning, the ones through which the fungi penetrate and at- Winetwork interviews.
tack the wood, compromising the integrity of the sap system
of the vine.
Practical application
Conditions of application Trunk cleaning consists in digging in the trunk to re-
move grapevine diseased parts. These damaged parts
The key point for applicability is the capacity of plant to reco- are often localized close to dead wood zones and below big
ver, that means it should not be too damaged and weak. In pruning wounds. First, the trunk need to be open where dead
general, practical experience recommends to avoid to apply wood has been detected, do not hesitate to wide open the
it on plants where the symptoms are too explicit and conso- trunk and take off dead wood. Then, detect diseased parts,
lidated. Disease wood symptoms need to be light, this recognizable by their spongy and toxins secretory form (also
technic is not effective against apoplexy. The earlier you called “amadou”). Once diseased part are detected, tissues
intervene, the better it is. Young vineyards show much bet- need to be removed by scraping using the side of the
ter results than old ones.This technique should be applied chainsaw in the grapevine axe and taking care to not cut-
on mature vines affected by Esca or Botryosphaeria dieback ting sap flow (Picture 1).
Other interesting point, there is no correlation between
severity of wood deterioration and severity of leaf
symptoms (Calzarano and Di Marco, 2007) and then on
the effect of taking-off white rot on symptoms foliar expres-
sion.
Some studies investigated which toxic metabolites were
produced by F. mediterranea, the basidiomycetous fungus
which is associated with grapevine white rot. These studies
reported that the culture filtrate of F. mediterranea produced
4-hydroxy-benzaldehyde, dihydroactinolide and a novel
chromanone, called 6-formyl-2,2-dimethyl-4-chromanone,
and biogenetically related to eutypine, in low concentrations
(ranging from 0.005 to 006 mg/L. (Tabacchi et al, 2000 ;
White , 2010). Not much is known about how these
metabolites act in the vine cells or tissues (Andolfi
et al, 2011).
Picture 1: Technique of trunk cleaning (IFV Alsace)
A trial made by the French Institute of Vine and Wine in
Recommendations :
Alsace in 2015 and 2016 on trunk cleaning showed that
Respect sap flux during pruning. Do not hesitate to well cleaning the trunk does not affect the vine response to wa-
clean out the wood in order to suppress of diseased ter stress and show no significant difference of yield (we
wood. If symptoms reappears, it is possible to realize a
second trunk cleaning during the year. can add that yield was a little bit higher for the cleaned
trunk than for control) and vigour is slightly better for the
cleaned trunks. Concerning GTDs expression, first results
Outcomes and empirical data on showed that cleaned modality (192 grapevines cleaned
effects and 178 grapevine of control) express less GTDs symptoms
than the control : 8.9% of symptomatic vines for the moda-
No more symptoms expression after the cleaning. The
lity with trunk cleaning and 15.7% of symptomatic vines in
cleaned grapevine is more vigourous than the non cleaned
the control (Gouttesoulard, 2016). These results need to be
vines and if the technique is done as soon as first symptoms
completed and the trial to be replicated in order to have a
appears (around July), harvest can be saved.
clear display of trunk cleaning effect on grapevine
trunk diseases expression.
Some scientific elements Another trial made in France by SICAVAC on trunk cleaning
since 2014 show good results, more than 600 grapevines
From the scientific point of view, basic element in pruning showing GTDs symptoms where cleaned and 700 grape-
techniques have solid evidence, nevertheless on the specific vines are used for control. In 2012, 8,7% of cleaned grape-
trunk cleaning concept there is still need for scien- vines express GTDs symptoms and 16% for the control, in
tific tests. What makes the assessment complicate is the 2015, 4,3% of cleaned grapevines express GTDS symptoms
need for long term trials, able to verify if the healing is and 14.2% for the control (Thibault M, 2015).
permanent or if symptoms appear again after a pe-
riod of time. Moreover, the physiological process the plants
undergoes after the cleaning is not clear.
Other requirements
We don’t know actually relationship between wood necrosis Time of cleaning: 100 to 200 grapevine for one day, around
and foliar symptoms expression. 5 minutes per grapevine ( 2 minutes for an expert).
We can ask the question: what is white rot and white rot Estimated cost: 2.5€/grapevine (Thibault M, 2015).
quantity incidence on foliar symptoms expression? Is there
one? White rot is created by Fomitiporia mediterranea that is When diseased vines are uprooted and substituted with new
deteriorating dead wood. shoots, the vineyard presents uneven quality in its grapes,
F. mediterranea cannot cause by itself white rot, it is a sapro- which affects the quality and the quantity of the grape pro-
phyte fungus. In order to cause this necrosis typical from duced. Tree surgery can have an economic consequences
esca, F. mediterranea need to colonise aleardy dead wood, for the producers, since it saves the costs of replantaing
for example healing cones, then necrosis is limited, or necro- (uprooting the diseased vines, digging the holes, implanting
sis created by other fungi as necrosis created by P. chlamy- the vine shoots, training), and does not imply a lack of pro-
dospora et P. aleophilum. duction from the new plants for at next three years.
TRUNK CLEANING
Concrete case: what happen on the field?

1- Trunk cleaning in south West
In South-West (France), this technique is not common, only

a very few winegrowers are applying it. Nevertheless, wine-
growers applying trunk cleaning are asking for more training
in order to be more efficient.
One example here of a winegrowers from Saint-Mont area in
Gers department.
Trunk cleaning is applied since 2014 on a plot of Cabernet
Sauvignon affected by esca and Botryosphaeria dieback.
Affected grapevine are identified in September, before har-
vest and marked. Then after harvest, grapevine is cutted
vertically with a small chainsaw in order to open the trunk.
Picture 2: Trunk cleaning made by a winegrower in South-West,
Once trunk is open, amadou (white rot) is localised and France (IFV South- West)
suppressed with the chainsaw. If the entire amadou is
not suppressed, grapevine can express the symptoms again.
It is then possible to clean the trunk a second time in order Estimated results:
to suppress all white rot (Pict. 2) As this is not a scientific trial, no clear numerous re-
Trunk cleaning also allow to identify pruning errors, to see sults are available. Nevertheless, winegrower impres-
sion is that after cleaning 90% of cleaned grapevine
dead tissus, sap flux and help the winegrower to better un- survived and doesn’t express symptoms anymore.
derstand pruning respecting sap flux. Winegrower add that it is too soon to conclude on the
According to the winegrower cleaning the trunk is taking real efficacy of this technique since it is used for two
around 5 minutes for one grapevine. years only.
2- Trunk cleaning in Italy
A small chainsaw is used to open the trunk and remove

the parts affected by the Esca disease, sometimes leaving
only the lateral parts to allow the sap to continue flowing.
Afterwards, with smaller saws the deteriorated wood under-
neath the cones of dryness is cleaned out (Pict. 3-4) . The
limited experience shows that after such operations more
than 80% of the plants did not show new symptoms of
necrotomy. The plant is then detoxed and strengthened and
can bear fruits at its full productivity.
Picture 4: How the surgery changes the plant (IPTPO, K. Diklić)
Estimated results:
Recent trials, performed by an Italian advisory com-
pany, gave promising results, leading to 97% of
asymptomatic plants in just one year. In 6 years 10,000
plants were treated in Italy and France. According to
the society, 90% of the plants that underwent treat-
ment have returned to full productivity. The plants are
under observation for the following years in order to
Picture 3: first part of the operation: opening the trunk and dertemrine long term results.
identify white rot (IPTPO, K. Diklić)
Scientific articles agronomique, de vinification et d’analyses du cente. Communication lors

des jounées nationales maladies du bois, 17 et 18 novembre 2015, Uni-
Andolfi A., Mugnai L., Luque J., Surico G., Cimmino A., Evidente A.,2011. versité de haute Alsace.
Phytotoxins produced by fungi associated with grapevine trunk diseases.
Toxins, Toxins, 3(12), 1569-1605.
Calzarano F., Di Marco S., 2007. Wood discoloration and decay in grape- More example in videos
vines with esca proper and their relationship with foliar symptoms. Phyto-
pathologia Mediterranea, 2007, 46, 96-101. http://www.winetwork-data.eu/fr/videos/aspects_pratiques_de_la_
lutte_contre_les_maladies_du_bois_de_la_vigne_sc_15728.htm
Gouttesoulard M., Experimental report France AgriMer, 2016
http://www.winetwork-data.eu/fr/videos/la_technique_du_curetage_
Lafon R., 1927. Modifications à apporter à la taille de la vigne dans les
pour_lutter_contre_les_maladies_du_bois_de_la_vigne_sc_15727.
Charentes. Taille Guyot-Poussard mixte et double. Roumégous et Dahan,
htm
Montpellier, 1921.
http://www.winetwork-data.eu/fr/videos/operation_curetage_sur_sau-
Larignon P., 2016. Etude des maladies cryptogamiques de la vigne:
vignon_blanc_pour_lutter_contre_les_maladies_du_bois_sc_15647.
symptomatology et agents pathogens. Institut français de la vigne et du
htm
vin, p 168.
Larignon P., Yobregat O., 2016. Cahier pratique: comment lutter contre
les maladies du bois de la vigne ? IFV, 7p.
Thibault M., 2015. Le Curetage. Service interprofessionnel de conseil

Thanks to Dr. Philippe Larignon and Dr.
Vincenzo Mondello who participated to
the develipment of this datasheet.
APPENDIX 10: PEROXYDE HYDROGEN INJECTION
www.winetwork.eu
21/27
HYDROGEN PEROXIDE INJECTION
Practice applied in 3 regions of Winetwork project, South

West of France, Galicia (Spain) and Douro (Portugal) using
different methods.
HYDROGEN PEROXIDE INJECTION
GENERAL PRINCIPLE
This techniques consist in one or more injection inside the trunk of grapevine of peroxide hydrogen in order to
limit symptoms expression. This technique is applied by winegrowers in South of France (Madiran), Spain
(Galicia) and Portugal (Douro). Application is easy and quick and results seems to be promising.
APPLICATION CASES
1- IN SOUTH-WEST
Injections were experimented in two plots characterized by the following:

Cultivar: Cabernet Sauvignon
Planting date: 2004
Rootstock: 3309 C
Pruning system: Guyot double
Yield: 60 hl/ha
Type of soil:clay and limestone
Pruning debris: pruning debris are crushed on the floor
Soil Management: natural grass cover between the rows. Chemical management of grass under rows.
No irrigation system.
Cultivar: Gros Manseng

Planting date: 2006
Rootstock: 3309 C
Pruning system: guyot
Yield: 30hl/ha
Type of soil: clay and limestone
Pruning debris: pruning debris are crushed on the floor
Soil Management: Chemical management of grass under rows, soil labour between the raw.
Application area:
In South West region in the area denominated “Madiran” close to Pyrenees mountains.
Climatic conditions are characterised by high annual rainfall around 1100 mm and 670 mm of rain during
vegetative period. Annual average temperature are comprised between 2°C minimum in winter and 25°C in
summer. The region also show a good sunshine of around 1400 hours of sun during vegetative season.
Page 1
Concrete application:
As soon as first foliar symptoms appears, diseased grapevine is marked. Rapidly the injection is done.
On the marked grapevine, make a drill with a driller, the best is to use a big wood wick. Drill need to be done
below vine arms (in the previous years some experiments were done on the location on the drill ; at the top of
the trunk, basis of the trunk, in one arm, below the arms ; the more effective appears to be below the arms).
In order to drill properly and to retain the liquid, give a inclination of 35 to 45° in the trunk axis. Once the drill
is made, inject with a serynge 3 to 4 ml of hydrogen peroxide (10%) in the drill. In average, winegrower spend
1 minute per grapevine to make the injection.
Winegrower is using this technique for 5 years on Cabernet-Sauvignon and for 2 years on Gros Manseng.
Over time, technique was refined and the most efficient entry point was found.
Page 2
2- IN GALICIA
The injection with H2O2 was applied in 3 vines of a plot of cultivar Albariño (white) with unless 15 % of
affection.
The plot is characterized by the following:

Cultivar: Albariño
Vine age: 30-40 years old
Non rootstock or also american rootstock of 1ª generation (V rupestris de Lot)
Training system: “Parral” (arbour)
Planting density: 5x4 m
Pruning system: Guyot.(Rod and spur pruning)
Yield: 10000 kg/Ha
Pruning debris: Infected pruning debris are taken out. Rest of them are crushed and left on the floor.
Soil Management: natural grass cover and mechanical management 3 times along vine cycle (winter, May and
pre- harvest)
Fertilization: Organic fertilizer, usually horse manure is distributed every year in winter with milling machine or
toothed cultivators (shanks).
Application area:
Area denominated “O Salnés” in SW Galicia close to sea cost and belongs to Rías Baixas Designation of Origin.
Atlantic climate: High annual rain fall: 1566 mm and 593 mm during vegetative period and 1400 hours of
sunshine.
Annual average temperature: 17.9 ºC. Between 4.7 ºC in winter and 27º C in summer and 1400 hours of sun
during vegetative period
Soil: clayey quite heavy.
Concrete application:
Previously vines were marked in summer, when foliar symptoms are clear
The application was done after pruning on third week of February in phenology stage 01 winter dormancy.
Each vine is 2 m high so it was made two holes with a drill (drill bit: 8) in two parts of vine: two at the base of
trunk and two at the top (1,8.m high).
It was injected the hydrogen peroxide with a syringe, and then he covered the hole with 2 cm3 of pure
eucalyptus oil. It was injected H2O2 30% p/v (110 vol) in three vines affected by Esca.
It was spent almost 3 minutes per vine including (drilling +injection both H2O2 and eucalyptus oil)
This practice was applied only one year and results are unknown so far.
The same winegrower will applied again this technique in other vines of the same plot.
Page 3
Ilustración 2 eucalyptus oil

Ilustración 1 Hydrogen peroxide
30% (p/v)
Ilustraction 3.- Drilling 8 mmm at the top and bottom of vine
Figure 1Injection Hydrogen peroxide 30%p/v (110 vol)
Page 4
SOME SCIENTIFIC ELEMENTS
First, it is known that H2O2 is a ROS (oxidative species of oxygen) that plant produces in responses to biotic or
abiotic actions. In the case of an attack of a pathogen, these oxidative species are fundamental in the defense
against fungal infection inducing enzymatic or non-enzymatic defense reactions of the plant.
Hydrogen peroxide can diffuse into the cell and activate the defense genes, leading to programmed cell death
(MCP) (Davison et al., 2002, Grant and Loake, 2000, Hammond-Kosack and Jones, 2000), which contributes to
limit the invasion of potential pathogens (Bokoch, 1994)
Correlation between ROS accumulation and the establishment of defenses suggests that ROS could have a
direct toxic function on the pathogens, or on the cells where they occur. The evidence also indicates that ROS
act as signals regulating the establishment of defense reaction at level of gene expression or the strengthening
of the cell wall by cross-linking of proteins from the wall (Apostol et al., 1989; Babior, 1992; Blechert et al.,
1995).
It has also been postulated that ROS participate in the induction of programmed cell death at the site of
infection, which contributes to limit invasion of potential pathogens (Bokoch, 1994). EROs may have opposite
functions in different plant / pathogen interactions, being in some cases positive regulators of defense and cell
death reactions, while in others they act as negative regulators (Montillet et al., 2005).
By the other hand, different trials to check several products against Esca, eutypa dieback and botryosphaeria
dieback in which grapevine rootstock and scion cuttings soaked in a product, like a Bio-Steriliser (hydrogen
peroxide) results were inconsistent. (Fourie & Halleen, 2006).
However, information about US patent “Method of protecting growing plants from the effects of plant
pathogens -US 6024986 A” which summary said “The direct introduction or injection of peroxy compounds
into a transpiration layer in a plant structure found inside the protective layer, bark or husk of a growing plant
is described as an effective method in treating plant disease and can achieve levels of protection unavailable in
the simple application of the materials to the leaf, stem, root or plant environment including air or soil”.
In the beginning of 2000’s some studies were conducted in order to evaluate the efficacy of fungicides in the
control of grapevine trunk diseases. Several fungicides were injected into the trunk with a high pressure pump
after drilling a hole with a drill. Injection were tested on several cultivars: Cabernet Sauvignon, Cabernet franc,
Riesling, Pinot, Gamay, Fer Servadou and Sauvignon. Any of the studies gave positive results, efficiency of
fungicides varying according to the region and cultivar, action seems to be limited in time, since as the second
or third year of observation percentage of diseased plants was similar to control (Sentenac et al., 2004,
Lecomte et al., 2006). Furthermore, the effect of an injected product and its mobility inside the trunk is
unknown.
Actually, other scientific elements are missing in order to understand and assess peroxide hydrogen efficiency
into the plant. We don’t know if sap is transporting peroxide hydrogen into plant vessels or if product stay at
injection point and either what is the response of the plant to this product and how product is affecting GTDs
pathogens.
Some winegrowers are using this technique on the field but are asking for further information and for more
research on this field in order to understand the effect of H2O2 on the plant and if H2O2 has a real efficiency
Page 5
on GTDs pathogens. Furthermore, questions on application are raised: what is the correct dose? What happen
if injected volume is higher? Does the plant die?
BACKGROUND
In Spain one winegrower of Galicia has applied this practice in 2016, in Castilla- León exists another
winegrower of Origin Designation Toro, who applies Hydrogen peroxide although with a different formulation -
HUWA SAN 50A (formulated on the basis of a synergistic mixture of hydrogen peroxide and colloidal ionic
silver). It has begun in 2015 with affected vines by Esca and results are being successful, although only one
year is few time to deduce favorable data..
The application is done immediately after pruning making an opening with a wedge, ax or with a drill, to reach
the heart of vine, and pour a 3% diluted solution (3L/100L water) into the opening of HUWA-SAN50A to
gradually penetrate into the open hole.Once the solution has been absorbed into the open holes, it is sprayed
on the whole plant and more specifically on the pruning wounds zone. In the following spring and when grapes
are on pea size (phenological state J-29 to K-31), it should be made a new directed spraying to canopy with this
compound at doses between 0.3 and 0.5% (300 to 500 cm3/ 100 l water). Through this new leaf spray, it is
intended to maintain the action of this product against Esca as well as preventing from other fungi and
bacteria
OUTCOMES
Injections made in South-West gave some results. It is important to remember here that it isn’t a scientific trial
but empirical approach. In 2015 the winegrower made the injection on 20 grapevines of Cabernet-Sauvignon,
the year after, none of the grapevine expressed GTDs foliar symptoms. In 2016 injections were made on 50
grapevines of Cabernet-Sauvignon. It appears that after injections symptoms are blocked and does not evolve
anymore and new healthy leaves appears. The grapevine seems to recover and don’t express the symptoms
the following years but more observations in the next years are needed in order to follow the treated plants
and see how they’ll evolve. On Gros Manseng, injections were done on 20 grapevines in 2016, and gave the
same results.
In Galicia, although the 3 vines showed symptoms on leaves and branches, only one of them seems to show
recovery because it had production although with smaller grains. In addition trunk and rest of branches show
wood in good condition once parts of dead wood have been removed.
It should be noted that these vines had already showed symptoms of Esca during the last 2-3 years and the
summer of 2016 has registered strong heat strokes with 38ºC of temperatures and vines have been bearing
water stress. The treatment will be repeated again in the vine that showed recovery and in more vines with
symptoms of the same plot in order to have a more representative number of treated vines.
More experimentation is needed in order to determine the real efficiency of hydrogen peroxide on symptoms
expression. In addition, residues analysis on leaves and clusters are necessary to determine if product is mobile
into the plant.
Page 6
Conditions of success Risks
AS soon as first foliar symptoms appears, Phytotoxicity
The sooner the better
Injection point below the arms Death of treated grapevine
Inclination of the hole Residues in clusters ?
On mature vines
INNOVATIVE ASPECTS
Only a few people are using this technique, probably mainly because there is no feedbacks on this technique
neither enough validated scientific elements. Nevertheless, it is noticed on field that more and more producers
are speaking of this technique, questioning, and some positive results are cited in wine press. On the field,
people applying injections of peroxide hydrogen are confidents even if some concerns are persisting.
OTHER REQUIRIMENTS
This practice can be appropriated easily, it can be done on all farm size. Of course, practice is time consuming,
so need to mobilize one person to realize the injections at a busy period. This technique don’t need any
particular training or background.
The bigger cost of this practice is labour cost (personal costs); in addition a driller is necessary, a bit and the
hydrogen peroxide that can be found in commercial shops.
W arning: using H2O2 is not accepted by the law, H2O2 is not a registered product for
grapevine and more particularly for grapevine trunk diseases.
SOURCE OF INFORMATION
Fourie, P.H. & Halleen, F, 2006. Chemical and biological protection of grapevine propagation material from
trunk disease pathogens. European Journal of Plant Pathology 116-255.
Foyer CH, Lopez-Delgado H, Dat JF, Scott IM, 1997. Hydrogen peroxide- and glutathione-associated
mechanisms of acclimatory stress tolerance and signalling. Physiologia Plantarum 100, 241–54.
Jochen Fischer, Stéphane Compant, Romain J. G. Pierron, Markus Gorfer, Alban Jacques, Eckhard Thines,
Harald Berger, 2026. Differing Alterations of Two Esca Associated Fungi, Phaeoacremonium
Page 7
aleophilum and Phaeomoniella chlamydospora on Transcriptomic Level, to Co-Cultured Vitis vinifera L. calli.
September 22, 2016, http://dx.doi.org/10.1371/journal.pone.0163344
Lecomte P., Darrieutort G., Liminana J.M, 2006. Maladies de dépérissement de la vigne : essais récents de lutte
au vignoble, méthodologie et résultats. 8ème conférence internationale sur les maladies des plantes, Tours, 5 et
6 décembre 2006, p876, 15-23
Sentenac G, Larignon P., Molot B., Viguès V., Kuntzmann P., 2004. Evaluation de l’éfficacité de fongicides et
d’agents biologiques utilisés contre les maladies du bois Esca et BDA. Premiers résultats d’expérimentations
menées sur le terrain. Les maladies du bois en Midi-Pyrénées, décembre 2004, 28-33.
Page 8
APPENDIX 11: USE OF MYCORRHIZAE IN FIELD
www.winetwork.eu
22/27
TECHNICAL
DATA SHEET
USE OF MYCORRHIZAE TO FIGHT AGAINST
GTDs
Mycorrhizae can be considered an essential factor in improving productivity

and plant health, since they contribute to improve the root system and,
therefore, the overall development of the plant.
There are some results that confirm the application of mycorrhiza as a
technology that favors the transplantation and subsequent development of
crops and provides a level of protection against adverse conditions.
Its effects could be especially interesting in a crop such as vine, which is
increasingly done with less phytochemical inputs and where quality and long-
term results prevail over the intensification of production.
In recent years experiences are being carried out using the application of
mycorrhiza as a support technique to improve the resistance of the vine plants
to the Grapevine Trunk Diseases, with non-definitive but hopeful results.

PRACTICE NAME
USE OF MYCORRHIZAE TO FIGHT AGAINST GTDs
DESCRIPTION
Mycorrhiza is the term used to describe the mutualistic association between the roots of the majority of
higher plants (both cultivated and wild) and certain microscopic soil fungi.
There are various types of mycorrhiza, however, we can say that arbuscular mycorrhiza are the most
important and widespread, since they are found in practically every climate, in all types of soil, in a large
number of plant species and formed by a greater number of fungal species. This type of mycorrhiza is
generally described using the abbreviation AM, although many countries use AMF.
The main benefits of mycorrhizae for the vineyard are:

• Increase of root system and volume of soil explored
• Increased efficiency in nutrient and water intake
• Higher resistance to water and saline stress.
• Increased tolerance to root pathogens (nematodes and fungi)
• Promotes balanced growth of grapevine
• Protects the root in its first phenological state
In the last years the use of mycorrhizae to help the plant to fight against GTDs is increasing. There are several
ways to use mycorrhizae, alone or in combination with Trichoderma species.
In this Technical Datasheet an experience carried out by one of the most important nursery in Spain, in
collaboration with several of its clients throughout the country, will be described.
Three ways of application of mycorrhizae are used:
Page 1
Gel:
The roots are introduced into the container until completely covered so that they are impregnated with the
product. A single application is enough for the entire life of the plant.
The mycorrhizal fungus germinates when the humidity and temperature conditions are adequate. Then the
fungus moves on the ground until finding the root and form the mycorrhiza. At this point the nutrient and
water exchange between both organisms is realized in a regulated and balanced way according to the needs
of the plant.
(Photo: Atens, SL)
Tablet:
Formulation (mycorrhizal fungus + Trichoderma spp. + NPK fertilizer) to apply in vines in manual plantations
and replenishments. The tablet is introduced into the transplanting pit at the time of planting. A single
application is enough for the entire life of the plant.
The fungus forming the mycorrhiza, and Trichoderma, germinates when the humidity and temperature
conditions are adequate. The mycorrhiza moves in the soil until finding the host root, where the exchange of
nutrients and water will be made between both organisms in a regulated and balanced way. In turn,
Trichoderma colonizes the rhizosphere of the plant displacing the possible pathogenic fungi improving the
health of the vine
Page 2
(Photo: Atens, SL)
Powder for irrigation:

Formulation to apply in vineyards by means of irrigation system in all type of lands. The product is poured
and dissolved in the irrigation tank. Can be applied in any vegetative state of the vineyard until two years
after its planting. A single application is enough for the entire life of the plant.
The mycorrhizal fungus germinates when the humidity and temperature conditions are adequate. Then the
fungus moves on the ground until finding the root and form the mycorrhiza. At this point the nutrient and
water exchange between both organisms is realized in a regulated and balanced way according to the needs
of the plant.
(photo: ICVV, LA Rioja (Spain))
The products used in this experience have the next composition:
Page 3
They are propagules of fungi that appear in the form of rootlets colonized by the fungus in addition to
microscopic structures of the same. Each gram of product contains a total of:
• Powder: 1400 spores of Glomus intrarradices and Glomus mosseae
• Tablet: 300 spores of Glomus intrarradicens, 1x106 conidia of Trichoderma koningii, 4x106
rhizosphere bacteria and NPK fertilizer: 5-5-3
• Gel: 100 spores of Glomus intrarradices and Glomus mosseae
The dose of employment of each formulation is:

• Gel: 1 kg / 150-250 grapevines
• Tablet: 1 tablet / grapevine
• Powder: 1 kg 2500 - 3000 grapevines
APPLICATION AREA
The vineyard where the experience has been carried out, is located in the town of Larraga (Navarra), within
the Denomination of Origin Navarra. The climate is semi-arid dry Mediterranean, with little precipitation
(approx 450-500 mm / year) distributed mainly in spring and autumn. The soils are varied, usually of
limestone type (marlstones, sandstones, ...) and often gravelly.
In addition to this experience, this type of products have been applied in more than 1000 hectares of
vineyard in Spain, through the collaborating clients of the nursery
RESULTS
Winegrowers who have tried this type of products report an improvement in budding and greater
homogeneity in the development of the plant. Users of tablets in replanting plants observe a higher rate of
growth.
Most admit that the effect is best when mycorrhizae are used in combination with Trichoderma.
However, these results are based only on practical observations, without sufficient support for scientific
results, which would require further studies..
SCIENTIFIC DATA
Page 4
Vineyard management protocols that can reduce the spread of grapevine trunk diseases are well covered in a
number of papers, including mycorrhizal additions to new plantings (Bleach et al. 2008), however this
technique has been shown to be inconsistent and other unpublished research suggests it has no value
beyond improving vine vigour in general (Jaspers, M 2011).
Arbuscular mycorrhizal fungi (Acaulospora laevis and Funneliformis mossea ) have been shown to increase
root dry weight and tolerance of grapevine rootstocks to black foot disease caused by Ilyonectria spp., and
changes in the function of the rhizosphere microbial community (Jones et al., 2014). Petit and Gubler (2006)
also indicated that grapevines inoculated with an arbuscular-mycorrhizal (AM) fungus, Glomus intraradices
were less susceptible to black-foot disease than non-mycorrhizal plants. This inoculation also reduced the
number of root lesions
INNOVATIVES ASPECTS
The use of mycorrhizae, although well known for some time, has not been used in general in Spain, and even
less linked to the fight against GTDs. It can’t be considered as a curative solution, but it could be included in a
program of preventive struggle, along with other cultural practices. In addition, by its characteristics, could be
suitable for an ecological production program. More experiences are needed to obtain more testable results.
OTHER REQUIRIMENTS
These techniques can be easily carried out on all types of farms, and have an approximate cost of:
• Gel format: € 105 / ha (product) + treatment cost (First year of planting)
• Powder for irrigation: 185 € / ha (product) + treatment cost (1-3 years after planting)
• Tablets: 0.10 € / vine (product) Replanting time
BIBLIOGRAPHY
Bleach, C. et al., 2008. Impact of mycorrhizal colonisation on grapevine establishment in cylindrocarpon

infested soil. New Zealand Plant Protection, 61, pp.311-316.
Gramaje, D. and Di Marco, S,. 2015. Identifying practices likely to have impacts on grapevine trunk disease
infections: a European nursery survey. Phytopathologia Mediterranea, 54(2), 313−324
Jaspers, M., 2011. Cylindrocarpon Black Foot Disease of Grapevines – Executive Summary (Personal
communication, 3rd October, 2011)
Jones, E.E., Hammond, S., Blond, C., Brown, D.S., & Ridgway, H.J. (2014). “Interaction between arbuscular
mycorrhizal fungi and rootstock cultivar on the susceptibility to infection by Ilyonectria species”.
Phytopathologia Mediterranea, 53(3), 582-583.
Page 5
Newsome, J., 2011. Grapevine Trunk Diseases: A review.
OIV, 2016. Grapevine Trunk Diseases: A review
Petit, E., & Gubler, W.D., 2006. Influence of Glomus intraradices on black foot disease caused by
Cylindrocarpon macrodidymum on Vitis rupestris under controlled conditions. Plant Disease, 90, 1481–1484.
Nogales, A., Aguirreolea, J., Santa María, E. et al., 2009. Response of mycorrhizal grapevine to Armillaria
mellea inoculation: Disease developement and polyamines. Plant and Soil, 317: 177-187
Nogales García, A., 2009. Estudio de la Interacción entre el hongo formador de micorrizas arbusculares
Glomus intraradicens Schenck y Smith y el hongo patógeno Armillaria mellea (Vahl:fr) P. Khun en vid. Tesis
doctoral. Universidad de Barcelona.
Nogales A., Camprubí Nieto A., Estaún Morell, V. Marfà V. y Calvet Pinós, M.V., 2010. In vitro interaction
studies between Glomus intraradices and Armillaria mellea in vines. Spanish journal of agricultural
research, ISSN 1695-971X, ISSN-e 2171-9292, Nº. Extra 1, 2010, págs. 62-68
MORE INFORMATION
www.winetwork-data.eu
Technical datasheet :
• Global vineyard strategy to prevent Grapevine Trunk diseases (GTDs)
Page 6
APPENDIX 12: COPPER NANOPARTICULES INJECTIONS
www.winetwork.eu
23/27
ñh
TECHNICAL DATA SHEET1
Cooper nanoparticles application
[Nanotechnology is already being used in agriculture for the treatment of

some plant diseases. Their use may increase the efficacy of commercial
pesticides and insecticides by reducing their amount of application at
significantly lower doses required for crops with the environmental
enhancement entailed.
The application of copper nanoparticles to fight against grapevine trunk
diseases (GTDs) is a practice that has been identified in a vineyard from Galicia,
in Spain. They are carrying out periodic applications (via trunk injection) of
aqueous solution of copper nanoparticles in 26 years old Albariño vines
damaged by Esca. They also perform foliar treatments, spraying of a mixture of
copper nanoparticles and an elicitor based on amino acids from porcine blood.
Results have not been observed yet, since 2016 has been the first year of
application. They will perform the applications at least two years more.]
1
6 or 8 pages In total including pictures
C O P P E R N A N O PA R T I C L E S A P P L I C AT I O N
DESCRIPTION
The application of copper nanoparticles (Cu NPs) to fight against GTDs is a practice that has been
identified as innovative, since it was detected in only one interview out of the 219 realized throughout the
10 regions that participate in the Winetwork project.
The application of Cu NPs is being carried out in vines of Albariño affected by Esca complex, and located
in a vineyard at Galicia (Spain).
Characteristics of the vineyard in which the practice is being carried out:
- Rootstock: R110 y 196-17 CL
- Cultivar: Albariño
- Age: 26 years
- Planting density: 1200 plants/ha
- Yield: 8000-9000 kg/ha
- Training system: parral (arbour)
- Pruning system: Guyot. Pruning is started in November and ended in February. Manual scissor is
used and each pruner carries a sprayer with diluted bleach (50:50). It is strictly ordered to
disinfect the scissors after pruning every vine. Pruning debris is grinded and removed. Pruning
wounds are protected with mastic immediately.
- All plants showing Esca symptoms are marked annually with rigorous control about their evolution
or possible uproot and renewal.
Figure 1. Foliar symptoms in vines cv. Albariño affected by Esca
Page 1
The practice is carried out as follows:

- 2 injections of copper nanoparticles. The first one at the bud break (April) and and the second one
at veraison (end of August): An aqueous solution of Cu NPs (15 mg/l) is directly injected (using a
syringe) in the trunk, doing previously a hole with a drill (bit 8). Then, the hole is covered with
cotton.
Figure 2. Injection of Cu NPs solution into a vine affected by Esca.
- During these 4 months, between April and August, 4 / 5 foliar treatments are performed by spraying
a mixture of Cu NPs (ratio 10:1000) and an elicitor based on amino acids from porcine blood (3:1000).
This practice was applied for the first time in 2016, and it is planned to continue for another two years, so no
results can be yet assessed.
APPLICATION AREA
The treated vineyard is located in Condado county, southwest of Galicia, in D.O. Rías Baixas. The region is
characterized by loam-sand soil with clay and river stones, coastal oceanic climate with mild temperatures
and abundant rainfall and no snow or frost.
SCIENTIFIC DATA
Copper has a special interest because, unlike other antimicrobial metals, it has a broad spectrum of action
against bacteria and fungi. It has been used extensively in the agricultural sector as a pesticide for thousands
of years. The efficacy of copper depends on environmental conditions, the concentration of copper ions and
the type of microorganisms. More in general, agrochemicals are usually applied by spraying. And sometimes,
Page 2
only a low quantity of the active ingredient reaches the target, due to leaching of chemicals, photolytic,
hydrolytic and microbial degradation. Therefore, to have an effective control, it is necessary to use a bigger
quantity of the active ingredient, with unfavorable effects such as water and soil contamination. The use of
nanotechnology in agriculture can increase the effectiveness of commercial pesticides and insecticides
leading to an environmental improvement. To date it is already used in the control of some plant diseases and
for the assimilation improvement fundamental nutrients by plants. In fact, nanoparticles can be used as novel
formulations of pesticides, insecticides and insect repellents by nanoemulsion or nanoencapsulation
techniques. Nanoparticles of silica, polyethylene glycol, silver, aluminum, zinc oxide and titanium dioxide have
been tested with promising results.For example, double-layer zinc-aluminum hydroxide compounds have
been used for the controlled release of chemical compounds that regulate plant growth. Yields have been
improved by the use of fertilizers incorporated in nanotubes and the release of nitrogen caused by the
hydrolysis of urea has been controlled through the insertion of urease enzymes into nanoporous silica
particles. Some publications indicate that zinc oxide nanoparticles used in a wide variety of crops such as
cucumber (Zhao et al., 2013), peanuts (Prasard et al., 2012), cauliflower, tomato (Singh et al.2013) helped to
increase the efficiency of zinc use in crops.
Cu NPs have attracted more attention in recent years, because of its physical, chemical, antimicrobial
properties, as well as for its abundance (Betancourt et al., 2013). In addition, copper has an important
biological role in the plant photosynthesis, although it not take part in chlorophyll composition. Therefore,
copper has great potential in the sustainable technological development. In relation to plant diseases, Cu NPs
were effective, in vitro, towards several fungi and yeasts pathogens such as Staphylococcus aureus,
Pseudomonas aeruginosa, Salmonella choleraesuis and Candida albicans, associated to serious diseases (Ren
et al., 2009) (Rupareli et al.,2009) (Ramyadevi et al., 2012)
The antimicrobial properties of copper is linked to its ability in accepting or donate electrons, that is to have
both high catalytic oxidation and reduction potential levels. In its oxidation state (Cu 2+), it is highly effective as
antimicrobial, due to the interaction with microbial nucleic acids, enzymatic active sites and cell membrane
components .
Scientific Literature:
• Begum, N., Sharma, B, Pandey, R.S. Evaluation of insecticidal efficacy of Calotropis procera and
Annona squamosa ethanol extracts against Musca Domestica. J. Biofertil. Biopestici. 2010, 1, 101-109.
• Betancourt, R. Reyes, P.Y., Puente, B., Ávila-Orta, C., Rodriguez, O., Cadenas, G., Lira-Saldivar, R.H.,
Synthesis of copper nanoparticles by thermal decomposition and their antimicrobial properties.
Journal of Nanomaterials. Volume 2013, Article ID 980545, 5 pages
http://dx.doi.org/10.1155/2013/980545.
• Cioffi, N., Torsi, L., Ditaranto, N., Tantillo, G., Ghibelli, L., Sabbatini, L., D’Alessio, M., Zambonin, P.G,
Traversa, E. Copper nanoparticle/polymer composites with antifungal and bacteriostatic properties.
Chem. Mater. 2005, 17, 5255-5262.
• Jeyaraman Ramyadevi, Kadarkaraithangam Jeyasubramanian, Arumugam Marikani, Govindasamy

Rajakumar, Abdul Abdul Rahuman, Synthesis and antimicrobial activity of copper nanoparticles.
Materials Letters 2012, 71, 114-116.
• Sadhucharan Mallick, Shilpa Sharma, Madhuchanda Banerjee, Siddhartha Sankar Ghosh, Arun
Chattopadhyay, and Anumita Paul, Iodine-Stabilized Cu Nanoparticle Chitosan Composite for
Antibacterial Applications 2012, 4, 1313−1323
Page 3
• Ren G, Hu D, Cheng EWC, Vargas-Reus MA, Reip P, Allaker RP. Characterisation of copper oxide
nanoparticles for antimicrobial applications. International Journal of Antimicrobial Agents. 2009, 33,
587–590.
• Rupareli JP, Chatterjee AK, Duttagupta SP, Mukherji S. Strain specificity in antimicrobial activity of
silver and copper nanoparticles. Acta Biomaterialia. 2008, 4, 707-771.
BACKGROUND
This practice is being carried out within the framework of a research project funded by national public
funding. The beneficiaries are the winery in which the practice is being carried out and a nanotechnology
company, which produces the Cu NPs and the elicitor under study.
OUTCOMES
The project started in 2016, so the trials have only been carried out for a year. The project will last for 3 years,
so it is still early to have results and make conclusions.

The key point of this practice is the putative ability of Cu NPs for moving through the plant vessels to stop the
GTDs.
In this practice there is a clear environmental benefit, due to the drastic reduction of copper quantity with the
use of nanoparticles.
Disadvantages:
- The current high price of the Cu NPs.
- Foliar treatments are not effective against other fungal diseases such as Powdery mildew and Downy
mildew.
Potential risks:
The potential risks that this practice could entail are the typical ones linked to any phytosanitary application,
since appropriate product dressing and proper application are required (dosage and cleaning of barrel). To
date, no adverse reactions have been observed in the plant after application, but it is convenient to be careful
with the dosage, as copper have phytotoxic effects.
INNOVATIVE ASPECTS
This practice is considered innovative because of the 219 interviews that were performed in the 10 European
wine regions, this type of practice was identified only in one winery.
The use of copper sulfate as a fungicide in viticulture is well known and widespread, but as we pointed out in
previous sections, the application of nanotechnology in agriculture is a more environmentally friendly
alternative for the specific case of insect and pest control than methods with synthetic agrochemicals, which
Page 4
have produced many environmental problems. Their use can increase the effectiveness of commercial
pesticides and insecticides by reducing their amount of application to the soil at significantly lower doses
required for the crops with the environmental improvement that implies. In addition to the lower
environmental impact, there are studies that have shown that MIC (minimum inhibitory concentration) of
bactericidal agent is lower for Cu NPs than for Cu2 + ions (Mallick et al., 2012), thus displaying a higher
antimicrobial efficacy of the Cu NPs against copper sulphate.
OTHER REQUIRIMENTS
This practice is equally applicable to other types of vineyards. There are no differences between varieties and
vineyards. The protocol is de same.
In terms of cost, it should be noted that the treatment with copper nanoparticles has a high cost. About 450
euros per hectare.
Doses of copper in the vineyard have a legal limitation, but nanoparticles allow to work with doses well below
those allowed (15 mg / l).
FA contact data
Paula Aldeanueva Potel
Facilitator Agent in Galicia (Spain)
email : paldeanueva@feuga.es
Phone: +34 986 469 110 - Ext 203
Mobile: +34 681 042 375
Page 5
APPENDIX 13: ORANGE OIL APPLICATION
www.winetwork.eu
24/27
Orange oil application on Scaphoideus
titanus young larval stages
Atypical practice applied in field to control Flavescence Dorée vector

Hot Water Treatment
Description Application area

Use of orange essential based product is a practice that was
During WINETWORK project 219 interviews were realized noticed during Winetwork interviews in two regions, Eger (Hun-
with winegrowers in 10 European winegrowing regions and gary) and South-West (France).
the facilitator agents - as an outcome of the interviews -
identified some cultural practices to fight Flavescence do-
rée (Candidatus Phytoplasma vitis). One of the innovative/
atypical practice is the application of orange essential oil
on the main vector of the disease, Scaphoideus titanus on
young larvae stages which is used in only a few WINETWORK
regions.
Principle
The principles of phytoplasma control are:
• the uprooting of infected vines
Figure 1: Area of application of orange oil as complementary
• the monitoring of vineyards and nurseries treatment against Scaphoideus titanus (signalled by red dot),
• insecticide control of the vector. Results from WINETWORK interviews
Treatment measures for vector control are highly recom-

mended, except for the nurseries and the infected areas Practical application
(under phytosanitary quarantine) where the applications are
compulsory.
Control measures can be a problem in organic vineyards and There are two registered products in Hungary containing
for wineries in the process of converting to organic agricul- cold pressed orange oil, called WETCIT and PREV-B2. The
ture and also in case of integrated pest management. From first one is originally registered as an adjuvant, which is a
the point of view of reducing pesticide use, an alternative highly effective surfactant designed for use with pesticides,
method to control the vector leafhopper - a natural com- plant growth regulators and nutrients requiring a wetting and
pound - the orange essential oil is used by winegrowers spreading agent. WETCIT assists in overcoming water repel-
as a substitute or a supplement to the insecticides. lence. At the same time PREV-B2 is a foliar fertilizer based
The main active ingredient of orange oil identified as a bota- on boron which is an essential micronutrient for plant growth
nical insecticide is the terpene D-limonene. and development and also orange extracts. In France several
products composed by orange essential oil are commercia-
lised as PREV-B2, PREV AM, LIMOCIDE and ESSEN’CIEL. The
Even if this practice is applied by some last 3 products are registered for grapevine use in control of
winegrowers there is no scientific evidence of the downy and powdery mildew.
efficiency of these ingredient on significantly
reducing Scaphoideus titanus populations. None of the mentioned products are registered for
grapevine insecticide use neither in Hungary nor in
France.
Though when applying these products according to wine-
growers’ observation (3 winegrowers are using this formula-
tion) have a side effect in the dehydration of the body of
the young larvae.
Recommendations when using orange oil based The fumigant action of peel essential oils of Citrus sinensis
products: var. pear (pear orange = PO) were evaluated by Ribeiro et
1. Start the applications at the beginning of May when al. against Bemisia tabaci biotype B. Regarding their effects
first larvae appears on oviposition, the Citrus oil showed concentration-response
dependence, reducing the number of eggs as the concentra-
2. Remove the suckers continuously because larvae pre- tion increased (Ribeiro et al., 2010.)
fer to stay on sucker shoots’ leaves The repellency and fumigant toxicities of the peel essential
3. Put product in tank mix in every application (fungi- oils of Citrus sinensis var. pear, C. sinensis var. mimo, and
cide, foliar fertilizer etc.) from April-May to the end of C. aurantium cultivated in northeast Brazil were evaluated
July against Tetranychus urticae and it was found that these es-
sential oils could be used to advantage for the control of T.
4. Increase the amount of water volume (from 400 l/
urticae (Araujo et al., 2010).
ha to 800-1000 l/ha) in connection with the develop-
ment of shoots (foliage)
The fumigant toxicity of some essential oils on adults of
5. Use 0,4 to 0,8 % dose rate stored-product pests, including Tribolium castaneum, Sito-
6. Accomplish canopy management (shoot positioning, philus granarius, Callosobruchus maculatus and Plodia in-
shoot thinning, hedging) as precise as possible terpunctella were investigated. According to this study the
essential oil of C. sinensis var. hamlin had good fumigant
7. Monitor the spread of leafhoppers positioning yel- toxicity on T. castaneum, S. granarius and C. maculatus
low sticky traps in the vineyards and if the number of S. (Mahmoudvand et al., 2011.)
titanus adult exceed the 4 adult/traps use insecticides
(conventional or organic farming). The essential oils of some aromatic plants and from peels of
lemon (Citrus limon) and orange (C. sinensis) were tested for
their insecticidal activity against the vine mealybug (Plano-
coccus ficus). According to this study no phytotoxic effects
Some scientific elements were observed on grape leaves treated with the citrus essen-
tial oils in laboratory conditions and moreover the essential
oils from citrus was more or equally toxic compared to the
It is important to highlight that there isn’t scientific confir- reference (paraffin oil) product (Karamaouna et al., 2013.)
mation about the efficacy of the two products on S. Fungitoxicity of the essential oil of Citrus sinensis on post-har-
titanus. But according to scientific studies the orange es- vest pathogens was investigated and it was found to be ab-
sential oil show a dehydration effect on different soft solute fungitoxic against the 10 post-harvest pathogens.
chitin shell insects as well as on some pathogens (downy Scanning electron microscopy was done to study the mode
and powdery mildew). of action of the oil in Aspergillus niger and it was observed
that treatment with the oil leads to distortion and thinning
Data on orange oil effect on other insects: of the hyphal wall and the reduction in hyphal diameter and
Citrus sinensis (sweet orange) essential oil was tested for its absence of conidiophores (Sharma et al., 2006).
potential for wheat protection against Oulema melanopus. ”As a plant-based natural product limonene and other mono-
The chemical composition of the essential oil was analysed terpenes might have use in pest and weed control in organic
and twenty-five compounds were identified in the oil among agriculture after phytotoxicity on crop plants and, effects on
which d-limonene was predominant component (89.49%). non-target soil animals and natural enemies of pest have
Direct contact toxicity assay showed the effectiveness of the been investigated.” (Ibrahim, 2001.)
essential oil against O. melanopus larvae causing the morta-
lity of 85% during 48 hours (Zarubova et al., 2014.).
Kumar et al. evaluated the insecticidal activity of essential oil It could be interesting for the future to evaluate the
of sweet orange (Citrus sinensis) against the larvae and pu- effect of D-Limonene on Scaphoideus titanus larvae
pae of housefly (Musca domestica). According to this study and if there is really one, research on how to take
significant activity of C. sinensis essential oil against larvae it into accound in a global strategy to fight Flaves-
and pupae of housefly was found, morover extreme dehydra- cence Dorée vector.
tion and surface distortion was observed on oil treated larvae
(Kumar et al., 2012.).
Araujo CP, da Camara CAG, Neves IA, Ribeiro ND, Gomes Ribeiro ND, da Camara CAG, Born FD, de Siqueira HAA In-
CA,de Moraes MM, Botelho PD Acaricidal activity against secticidal activity against Bemisia tabaci biotype B of peel
Tetranychus urticae and chemical composition of peel essential oil of Citrus sinensis var. pear and Citrus aurantium
essential oils of three citrus species cultivated in NE Bra- Cultivated in Northeast Brazil. Nat Prod Commun 5:1819-
zil. Nat Prod Commun 5:471-476. doi:10.1590/S1519 1822 (2010)
566X2011000300011(2010)
Sharma N, Tripathi A Fungitoxicity of the essential oil of Citrus
Ibrahim MA, Kainulainen P, Aflatuni A, Tiilikala K, Holopainen sinensis on post-harvest pathogens World J Microbiol Bio-
JK Insecticidal, repelnt, antimicrobial activity and phytotoxi- technol 22: 587. doi:10.1007/s11274-005-9075-3 (2006)
city of essential oils: with specials reference to limonene and
its suitability for control of insect pests. Agr Food Sci Finland Zarubova L, Kourimska L, Novy P, Zouhar M, Douda O, Sku-
10:243–259. doi:10.1603/029.102.0422 (2001) hrovec J Botanical pesticides and their human health safety
on the example of Citrus sinensis essential oil and Oulema
Karamaouna F, Kimbaris A, Michaelakis A, Papachristos D, melanopus under laboratory conditions Mendelnet (2014)
Polissiou M,Papatsakona P, Tsora E Insecticidal Activity of
Plant Essential Oils Against the Vine Mealybug, Planococcus
ficus J Insect Sci. 13: 142. doi: 10.1673/031.013.14201
(2013)
More information on Flavescence Dorée on
Kumar P, Mishra S, Malik A, Satya S Insecticidal evaluation
of essential oils of Citrus sinensis L. (Myrtales: Myrtaceae) www.winetwork-data.eu
against housefly, Musca domestica L. (Diptera: Muscidae). Technical datasheets :
Parasitol Res 110:1929-1936. doi:10.1007/s00436-011-
2719-3 (2012) • Guide of good practices in FD management
Mahmoudvand M, Abbasipour H, Basij M, Hosseinpour MH, • How to be more efficient in FD management
Rastegar F, Nasiri MB Fumigant toxicity of some essential oils
• Hot Water Treatment
on adults of some stored –product pests. Chilean J Agric Res
71:83-89 (2011)
Work realized in common by the facilitators

agents of Winetwork project. Data came from
practice, through the help of 219 interviews
and from a review of scientific litterature.
Thanks to, Dr. Mauro Jermini who participated
APPENDIX 14: HOT WATER TREATMENT
www.winetwork.eu
25/27
Hot Water Treatment (HWT)

Prevent from propagation of Flavescence Dorée phytoplasma

Hot Water Treatment
Introduction Conditions of application

Flavescence Dorée (FD) is a type of grapevine yellow and Duration of treatment
a serious disease caused by phytoplasma. FD is spread by Treatment duration is from 45 to 65 minutes according to
an insect vector, a leafhopper Scaphoideus titanus, and it temperature. According to the country, diferent times and
has been shown that it can be transmitted by using infected temperatures are used:
propagation material, scions and/or cuttings collected from
diseased plant during a period of symptom latency. Hot water • 45 min at 50°C (Mannini et al 2009; Caudwell et al,
treatment of propagation material could prevent from this 1990; Caudwell et al, 1997; Tassart-Subirats et al, 2003).
danger. These parameters allow to eliminate phytoplasma
and have a partial effect on Scaphoideus titanus
Principle eggs (presents under bark on the one year old plant
material) (Caudwell et al, 1997; Linder et al, 2010).
Hot water treatment is proposed to cure dormant wood
material from phytoplasmas and to supress surface • 65 min: 25-35°C during 10-20 minutes, then 50°C
parasite and pests. Pathogenic agent, the phytoplasma is during 45 minutes. The first bath allow to prepare
heat-sensitive. Time and temperature need to be such as wood cuttings by soaking tissues at the real treatment of
supress phytoplasma without affecting plant develop- 50°C during 45 minutes (Piano S. and Costa C., 2017).
ment capacity. Hot water treatment was proposed first in
1966 by Caudwell to cure dormant woody plant material
from phytoplasma (both FD and Bois Noir). Several scien- Operational process
tific experiences have demonstrated the effectiveness of Hot water treatment need to be realized at the step of sto-
this treatment against pathogens (Caudwell et al, 1990; rage in cold room, before grafting (for scions) or before plan-
Tassart-Subirats et al, 2003; Mannini et al, 2009). Soaking ting (for grafted rootlings).
plants into hot water is an essential additional treatment Material need to be prepared before treatment by accli-
to limit the distribution of FD (and Bois Noir) by gua- mation at room temperature 12 hours before, then put
ranteeing its sanitary quality. back in store room with high humidity.
Do not make fungicide treatment in the same time as hot
Objective water treatment. If necessary, insecticide treatment can be
done after the HWT.
Eliminate Flavescence Dorée phytoplasma from plan- If the treated material is transported, use aerated packaging
ting material in order to limit inoculum propagation without with good hydration and controlled temperature.
affecting vegetative development.
Recommendations:
Use adapted material and immerge dormant cuttings and

grafted rootlings into a hot bath for 45 minutes at 50°C.
Using other parameters of time and temperature
can induce symptoms expression, lack of vita-
lity of treated plants. Store cutting material before hot
water treatment in cold room at 5°C with high humidity
and right after treatment, store again in cold room (Piano
S. and Costa C., 2017; GTNFD, 2006). Control efficien-
cy of the equipment and in particular the possibility of a
pre-heating of the material prior to the proper HWT to
prevent termic shocks, and the continuous and regular
flowing of the hot water around the wood cuttings.
IFV South-West
and haven’t suffer from any desiccation or reserves losses.
Outcomes Pay attention to varietal sensitivity, (due probably to a greater
or lesser diameter of the shoots, the marrow more
Plants treated with hot water treatment at 50°C for 45’
or less expanded, different content of reserve substances).
showed after one year vitality up to 75% (Mannini et al,
2009) and no FD symptoms. On the contrary, treatment at
• Reliability of treatment material
52°C for 45’ can induce as a side effect almost 10% reduc-
tion of take (Mannini et al, 2009). Temperature inside the bath need to be kept constant and
Hot-water treatment has also an effect on Bois Noir phytoplas- homogenous (probes are controlling temperature in the bath)
ma (treatment at 52°C), partial effect on leafhopper eggs, with an accepted variation of +/- 0.5°C (GTNFD, 2006). It
thrips, bacterial diseases (Agrobacterium vitis and bacterical is important to renew frequently water, according to treat-
necrosis, Xylophilus ampelinus) (Hamilton R., 1997; GTNFD, ments frequency. A factor determining operation success is
2006) and allow to clean from many parasites (Piano S. and the control of time and temperature combination, monitored
Costa C., 2017). during treatment.
Effectiveness of HWT is linked to material age, effectiveness
is better on one year old planting material.
IFV South-West
• Operational process
Soaking plant into hot water has an effect of thermic choc
that can modify physiological state of propagating material.
The above operation need to be respected in order to avoid
any problems of bud-burst and mortality.
• Prohibition of circulation of phytoplasma-infected

material
FD is a quarantine disease and might be dangerous for other
phytoplasmas, since potential vectors may exists in the area
IFV South-West of introduction.
Key points for success / risks Purchasing Hot Water Treated nursery material is strongly
recommended to European growers, in particular for areas
HWT complete insecticides treatments action, sup- where FD is not present yet. In presence of the vector, a
press state standing carriers of contaminated rootstocks, single infected plant can generate a wide infection.
supplement absence of chemical treatment against
phytoplasmas. In order to insure treatment efficiency and Combination of time and temperature is the most
preserve propagating material several conditions need to be important factor in HWT efficiency. If combination of 50°C
respected: for 45 minutes is not respected, buds can present localized
cellular degeneration and total alteration up to 60°C.
• Quality of propagating material
Wood cuttings need to be lignified and present the best pos- HWT at 50°C does not no influence damage of the conduc-
sible reserves. Cuttings should have been stored in the best tive tissues, or to any significant disturbances of hydraulic
conditions of temperature and hygrometry after picking time conductivity degeneration are observed (Remolif et al, 2014).
Mannini F., Argamante N., Gambino G., Mollo A., 2009. Linder C., Schaub L., Klötzli-Estermann F., 2010. Efficacité
Phytoplasma diffusion through grapevine propagation ma- du traitement à l’eau chaude contre les œufs de Scaphoi-
terial and hot water treatment. Progrès agricole et viticole, deus titanus, vecteur de la flavescence dorée de la vigne.
2009, Hors série – Extended abstracts 16th meeting of ICGV, Revue suisse Viticulture, Arboriculture, Horticulture, vol 42
Dijon, France, 31 Aug-4 sept 2009, 182-183. (2), 132-135
Caudwell A., Larrue J., Boudon-Padieu E., McLean G.D.,
1997. Flavescence Dorée elimination from dormant wood
of grapevines by hot-water treatment. Australian Journal of More information on Flavescence Dorée on
Grape and Wine Research 3 (1), 21-25. www.winetwork-data.eu
Caudwell A., Larrue J., Valat C., Grenan S., 1990. Les traite-
ments à l’eau chaude des bois de vigne atteints de la Flaves-
cence Dorée. Progrès agricole et viticole 107 (12), 281-286
Piano S. and Costa C., 2017. La termoterapia in acqua calda

come Sistema di lotta al fitoplasmi della vite. “Manuale per la
lotta ai fitoplasmi della vite”, edito da Provincia di Asti, 2017.
Groupe de travail national Flavescence Dorée, 2006. Jau-
nisses à phytoplasmes de la vigne, rapport, 24p.
Remolif E., Zufferey V., Dubuis P.H., Voinesco F., Fendeleur O.,
Gindro K., 2014. Traitement des bois à l’eau chaude contre
la flavescence dorée : effet sur l’anatomie et l’intégrité des
tissus conducteurs. Revue Suisse de viticulture, Arboricul-
ture, Horticulture, vol 46 (5), 302-308.

Thanks to, Dr. Mauro Jermini who participated
APPENDIX 15: WILD VINES MANAGEMENT
www.winetwork.eu
26/27
Wild American vines management
Draft by Maurizio Gily, 19/01/2017
The problem of uncultivated land
The flavescence dorée disease is more

dangerous in the not-wealthy wine-
growing areas, where some vineyards
were left to wild in the past decades, and
where viticulture is not intensive but
share the land with forests and
uncultivated.
The reason for this "social" and
territorial character of the disease is
related, in part, with the presence of FD
outbreaks sites outside of cultivated
vineyards, which are in some cases
recently abandoned vineyards, where no
one implements the compulsory
prophylactic measures, or, more often,
areas that were vineyards in the past and
today are bush or forest.
In these areas the European vine don’t
usually survives, but the American vine
rootstock, resistant to fungi, has
developed and, according with its nature
of liana, climbed on trees up to great
height forming a dense canopy.
Picture 1: big wild American vine

canopies at the edge of a field
American vines (picture 1) host both the disease and the vector.
The various crosses of American vines (Vitis rupestris, riparia, berlandieri etc.) and the ones between them
and European vines, used as rootstocks, are capable of hosting the FD phytoplasmas, although they
generally do not exhibit the typical symptoms of cultivated vine, or don’t show any symptoms at all:
therefore they are healthy carriers.
The vector Scaphoideus titanus feeds indifferently on all species of the genus Vitis, so he can acquire the
phytoplasma and transmit the infection from wild to cultivated vines. This is definitively established.
On wild vine in the refuge areas, where the insect is not disturbed by insecticide treatments, higher
populations were found, compared with that in vineyards in the neighboring areas. And many of them are
infected.
Picture 2 and 3: Scaphoideus in the vineyard and in the wild, percentage of infected
adults. Marzachì and Bosco, 2016
The flight of Scaphoideus
It has been proved (Alma, Lessio) that the adult insect vector is able to make quite long flights. However,
the greater the distance of the "wild" outbreak from the vineyard, the lower is the probability of a
migration into the vineyard itself.
The migration of insects from outside towards inside a vineyard is also demonstrated by the greater
frequency of the presence of symptoms at the border of the vineyard.
Picture 4
Pavan at al., 2012
The owner's responsibility
It’s then established that the owner of a plot with wild American vines creates damage to nearby vineyards
and to the whole territory, encouraging the preservation and propagation of the disease.
The discipline to force mandatory control in various states provides that the owner be requested to destroy
the source of inoculum, i.e. of American vinestocks. In case of default there are orders, sanctions, until
forced uprooting.
However, laws protect private property, so no coercive step is easily applied: it requires long bureaucratic
procedures and expenses, that often the public administrations are not able to support.
This is a very serious problem that requires clever solutions on behalf of the states!
In any case, it is usually recommended that those who own vineyards nearby Scaphoideus refuge areas look
for friendly agreements with the owners, to carry out the remediation, and only if they haven’t success ask
for law protection.
How to clean up?
It is not necessary to deforest. The vine is a sun-loving plant, looking for sunlight and then its canopy grows
mainly on the edge of forests, at the border of cultivated fields and roads.
Going further into the bush often can be not found any more.
Picture 5. Wild American vines along a road

But If it is allowed to grow and develop big canopies, it
goes to ripen fruit, and at that point can also be
propagated by seed, through the birds. This should be
avoided.
Picture 6. Bunches of wild American vines.

Birds can propagate by seeds
Cleaning in winter time
After the harvest, since late autumn, when the adult insect does not fly any more, until vine budburst:
cleaning of fallow stripes bordering the vineyard.
Operate with mulchers with telescopic arm, forestry mulcher; pay particular attention to the cutting of wild
vines climbing on trees.
The vine is a sun-loving plant, after the first few meters from the perimeter of the bush often can not be
found anymore. But if there is the presence, and using mechanical equipment is not possible to preserve
the presence of tall trees, the American vines must be cut by hand, for a buffer zone as large as possible,
but at least some ten of meters.
The debries must be removed and shredded on site or left hanging from trees. Laying on the ground they
could sprout from the cuttings and multiply. They can contain Scaphoideus eggs. At the time of hatching
the larvae will look for young vine leaves. They must not find them!
Where possible, eradicate the roots of the vine with an excavator.
Cleaning in springtime
May-June: before treatment against the youth in the vineyard, destroying sprouts of survived American
vines is required. As an alternative, spray localized herbicides with a high concentration of glyphosate (from
2 to 5% solution of the commercial product with 360g/L of glyphosate) in order to kill for starvation any
young specimens of the vector.
CAUTION: Do not destroy wild American vines in summer, because the adults may migrate from the wild
to the vineyard.
The procedure must be repeated in subsequent years, trying to eliminate all American vinestocks.
APPENDIX 16: MATING DISRUPTION
www.winetwork.eu
27/27
Mating disruption by using vibrational

signals
A new method for Scaphoideus titanus management

Mating disruption
In Europe, Flavescence dorée is a quarantine disease and

Introductory note there are compulsory measures to manage vector popula-
tions and prevent the spread of the disease which include
During WINETWORK project, 219 interviews were conduc- large-scale insecticide treatments. To apply the mandatory
ted by the facilitator agents (FA’s) of the 10 European wine- control methods, growers may need to apply large amounts
growing regions involved on the project. The FA’s identified of pesticides, which have both environmental and econo-
several practices to fight “Flavescense Dorée” phytoplasm mic impacts. Considering that there is an increasing mar-
and/or its vector, the cicadellidae Scaphoideus titanus Ball, ket demand for pesticides-free products (organic market),
from which one was the Mating Disruption using Vibra- and large food retailers are imposing more stringent limits
tional Signals to disturb the behaviour of S. titanus. than those in current legislation on residues, the EU Directive
Unfortunately, this biotechnical method is not yet available 128/2009, for the sustainable use of pesticides, is moving
commercially for winegrower’s application. However, since in the direction of finding alternatives to chemicals, the de-
this is a very innovative and promising tool, and there is a velopment of environmentally friendly alternatives is crucial.
strong interest of the winegrowers and technicians on this Vibrational Mating Disruption (VMD) is an example of an
technique, the consortium decided to create a technical da- innovative method, since it exploits vibrations used by this
tasheet to disseminate this technique. insect, both for mating and rivalry. Pair formation in S. titanus
is characterized by a continuous duet between a male
Introduction and a female that starts with partner reciprocal identifica-
tion and proceeds with a location (search) stage before the
Scaphoideus titanus Ball (Hemiptera: Cicadellidae) is a final courtship that precedes mating. The mating duet is cha-
leafhopper, the main vector of the phytoplasma that causes racterized by specific temporal pattern which change during
“Flavescence Dorée”, the most threatening among European the process of pair formation. In the identification duet, male
grapevine yellow diseases. Despite its importance, very little pulses are delayed after female reply, while they are fully
is known about this species besides its general biology and synchronized during location and courtship duets. One hypo-
mechanism of disease transmission and communication. thesis is that VMD is more successfully if applied during
In order to find alternatives to conventional pesticides, ge- the identification stage when external interferences could
nerally used against this pest, our attention needs to focus result in loss of important information that is needed to
on some behavioral aspects, in particular, to the mating be- correctly identify the mating partner. However, given the
havior of such species. Mate recognition and localization in complexity of the mating duet structure, interference caused
Auchenorrhyncha (with the exception of most cicada) are by VMD could affect each stage of the mating process. The
mediated via acoustic signals, transmitted through the subs- key to prevent S. titanus mating is the interruption of the
trate. It has been estimated that 150 000 species use vibra- sound vibrational sexual communication by transmit-
tions to achieve mating and among them there are several ting suitable disrupting signals.
pests and important vectors of plant diseases.
Figure 1: S. titanus Ball (Cristina Carlos, ADVID and Flavescence Dorée symptoms (Eng. José Freitas, DRAPN)
Males call spontaneously, carrying out a specific “call and
Methodology and application fly” behavior followed by a well-structured courtship song.
Females emit signals only in response to males and rival
Current knowledge about the sexual behavior of S. titanus males compete for mating, producing a disruptive noise ai-
and VMD experiments (Mazzoni et al. 2009a; Eriksson et al. med at interrupting a duet in place between pairs.
2012; Polajnar et al. 2014) suggested that playback of vibra-
tional disturbance noise (VDN; based on vibrational signals
used by males during rivalry) should suffice to disrupt the
courtship behavior and prevent mating, thus represen- Scientific data and some
ting a promising avenue for developing a non-chemical ap- results
proach for controlling this invasive vector in Europe.
After initial studies on the specie’s mating behavior, attention
Principle of the mating disruption protection method was focused on the possibility of achieving mating disrup-
for S. titanus. tion by playback of vibrational signals. Efficacy of playback
An electromagnetic shaker was used to vibrate the wire with with sufficient amplitude was first demonstrated in labora-
VDN that was transmitted to the plants. It was possible by tory trials, and then in semifield conditions with insect pairs
testing transmission of male calling signals on different plant placed in cages in an experimental vineyard. The approach
parts, establishing the active space of mating signals in or- was to gather knowledge of basic reproductive biology
der to adjust the power of the mating disruption signals into first, which revealed a naturally occurring disturbance si-
effective species-specific masking signals (VDN) (Mazzoni et gnal emitted by rival males (VDN) that masked the temporal
al., 2009b; Polajnar et al., 2014). structure of mating calls in antagonistic interactions between
males. The use of VDN has a distinct advantage over synthe-
tically generated pure tones waveforms, because its features
have evolved for efficiency, so spectral features are expected
to be optimal for this function. Although S. titanus is one of
the few species known to use acoustic disruption, masking
the vibrational signals, which are important for mate recogni-
tion, should be effective in other species as well.
Mazzoni et. al, in 2009 found that the playback of VDN re-
duced the level of male calling and interrupted established
male-female duet that consequently resulted in a signifi-
cantly reduced number of copulations. These results indi-
cate that the vibrational communication channel is open to
interference both from abiotic environmental noise and from
Figure 2: Shaker designed to transfer disruptive noise to longer dis- signals produced by sexual competitors or even heteros-
tances. (Source: Lucchi et. al)
pecifics. The study also suggests that a detailed unders-
tanding of a leafhopper behavior is essential for trying
new approaches in the development of more environmentally
friendly control practices.
Figure 3: Male calling signal; Courtship song/Mating duet; Rivalry disturbance noise of S. titanus (Source: Mazzoni et. al, 2008)
Mating disruption
Eriksson et. al (2012), showed, for the first time, that effec- In 2013, the same investigators applied the methodology to
tive mating disruption based on substrateborne vibrational longer distances. They found that is still possible to achieve
signals can be achieved in the field. When disruptive vibra- 65% of mating disruption at 45 meters with 18 hours of
tional signals were applied to grapevine plants through a shaker operation. They succeed in 80% of cases. The results
supporting wire, mating frequency of the leafhopper pest S. indicate that the principles from which the mating disruption
titanus dropped to 9% in semifield conditions and to 4% in with vibrational signals was validated is applicable even at
a mature vineyard. The underlying mechanism of this envi- field level, on mature plants. The disruption has been ef-
ronmentally friendly pestcontrol tactic is a masking of the fective on more than 90% of tested pairs when some condi-
vibrational signals used in mate recognition and location. tions were respected. In particular, they found that mating
In 2012, Lucchi et. al, in open field conditions, found signi- is almost totally prevented, when the device is working for
ficant differences between vibrated and non-vibrated (C) periods of more than 19 hours. Indeed, they detected im-
grapevine plants treatments, concerning the number of vir- portant losses and dispersion of the signal, due to numerous
gin females, using an electromagnetic shaker to vibrate the points of contact between vibrating wires/plants/poles. This
wire with disruptive signal. limit must be eliminated or strongly reduced, for instance by
passing the signal or using dampers in correspondence of
such critical points.
Polajnar et al. (2016), also published about using artificial

vibrational noise to prevent mate recognition and localization
mediated by vibrational signals in the grapevine pest S. tita-
nus. Building on the proof of concept published previously,
mating trials were set up in laboratory to determine the am-
plitude threshold for playback efficacy and reveal the mecha-
nism of its function, while field trials were performed to va-
lidate this threshold and explore the possibility of reducing
energy use by exploiting the diel pattern of this species’ ma-
ting activity. The threshold obtained in laboratory trials—15
μm/s peak amplitude—was confirmed by measurements of
attenuation and insect mating in field cages at successive
Figure 4: Number of virgin females found on vibrated and non-vibrated (C) grapevine plants in a field trial with mature grapevine
plants in 2012 (Distance = distance to control (c) modality).
distances from the source. They also discovered that shut-
ting off the disruptive noise between 1000 and 1800 h did
not reduce efficacy of the method in the field, allowing ener-
gy saving in this period. The noise had an allor nothing effect
on S. titanus mating behavior, and were unable to ascertain
the exact mechanism of the communication breakdown, but
the approach appears robust enough to merit largescale tes-
ting in the future.
Key Points for successful Conclusions

implementation of the The researchers believe that the use of acoustic devices in a
MD method sustainable way for growers is still to come, but the biological
knowledge to make it work and the technology are already
The adoption of strategies based on acoustic tools would available. The lack of solutions would be overcome if more
enable medium to long-term reduction in the use of che- directed efforts were made to unify and optimize knowledge
mical pesticides, which fits well within the IPM and organic already available and to study and develop new solutions
concepts. However, in order to develop a technique that will for practical application, according to the peculiarities of
be adopted by the public, it must become accessible and any crop-pest system where an acoustic based approach is
commercially viable. Such tools should therefore be (eco- feasible. However, it should be emphasized that many open
nomically) competitive with other solutions already available questions remain to be answered, such as the struggles in
on the market, namely considering the cost of the device the energy supply in the vineyards, before the method is
(purchase + maintenance). Equipment costs, as deemed ready for implementation, both technical and strategic.
feasible by the industry, could be brought to within 300 €/ha, Aside from the above-mentioned issues of attenuation and
which is comparable to the cost of pesticide treatments if a effect of vibrations on plants, there is plenty of room for im-
5-year lifetime of a unit is assumed. Another feature of such proving design and usability of the devices, which are still in
a system is the suspension of standard wires along the rows the prototype stage of development. Dealing with different
in vineyards, which can be used to deliver vibrational energy trellising systems (including those without supporting wires)
to individual plants, without the need for elaborate technical and abandoned vineyards should be carefully considered
solutions. as well, although these are the issues largely shared with
However, it´s important to understand better the effect of conventional mating disruption methods, so starting from
different environmental conditions (rain, atmospheric pres- scratch will not be necessary. That said, the researchers be-
sure change), and also the collateral effects on beneficial lieve that this approach is worthy of being pursued further
insects or other pests. and has potential to replace or at least supplement chemical
measures for control of S. titanus in Europe.
A.Lucchi, A. Eriksson, G. Anfora, M. V. Doberlet, V. Mazzoni, 33, Issue 2, Pages 174–185.
2013. A ten-year research on vibrational communication in Valerio Mazzoni, Andrea Lucchi, Janez Prešern and Meta Vi-
Scaphoideus titanus: science fiction or future prospect? rant-Doberlet, 2008. Vibrational communication and other
Ard Nieuwenhuizen, PURE - Pesticide Use-and-risk Re- behavioural traits in Scaphoideus titanus. Bulletin of Insec-
duction in European farming systems with Integrated Pest tology 61 (1): 187-188.
Management, 2007-2013. Collaborative Project, SEVENTH Valerio Mazzoni, Janez Prešern, Andrea Lucchi and Meta Vi-
FRAMEWORK PROGRAMME. rant-Doberlet, 2009b. Reproductive strategy of the Nearctic
Eriksson, A, Gianfranco Anfora, Andrea Lucchi, Francesco leafhopper Scaphoideus titanus Ball (Hemiptera: Cicadelli-
Lanzo, Meta Virant-¬Doberlet, Valerio Mazzoni, 2012. Ex- dae). Bulletin of Entomological Research, Volume 99, Issue
ploitation of Insect Vibrational Signals Reveals a New Me- 4, pp. 401 -413.
thod of Pest Management. PLOS ONE 9(6): e100029. doi: Veronelli V., 2009. MATING DISRUPTION IN IPM: pheromones
10.1371/journal.pone.0100029. and beyond. Lucerne 19-20 October, ABIM 2009.
Eriksson, A., 2013. Mating disruption in Scaphoideus tita-
nus Ball (Hemiptera: Cicadellidae) by vibrational signals. PhD
Thesis. University of Pisa Fondazione Edmund Mach.
Jernej Polajnar, Anna Eriksson, Andrea Lucchi, Gianfranco More information on
Anfora, Meta VirantDoberlet, Valerio Mazzoni, 2016a. Mani- www.winetwork-data.eu
pulating behaviour with substrateborne vibrations – potential
for insect pest control. Pest Management Science. Volume Technical datasheets :
71 , Issue 1, Pages 15–23. • Guide of good practices in FD management
Jernej Polajnar, Anna Eriksson, Meta Virant-Doberlet, Valerio • How to manage with more precision FD
Mazzoni, 2016b. Mating disruption of a grapevine pest using • Hot Water Treatment
mechanical vibrations: from laboratory to the field. J Pest
Sci. Volume 89, Issue 4, pp 909–921. Video seminar: State of the art of scientific research on
Flavescence Dorée (François-Michel Bernard, IFV)
Valerio Mazzoni, Andrea Lucchi, Andrej Čokl, Janez Prešern,
Meta VirantDoberlet, 2009a. Disruption of the reproductive Experiences of collective management of FD in France
behaviour of Scaphoideus titanus by playback of vibrational (François-Michel Bernard, IFV)
signals. Entomologia Experimentalis et Applicata, Volume 1

Thanks to Valerio Mazzoni who participated to
the development of this datasheet.

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Deliverable 2.5: TECHNICAL DATASHEETS IN INNOVATIVE SOLUTIONS TO CONTROL

INSTITUTO GALEGO DE CALIDADE ALIMENTARIA- INGACAL-

MARÍA JESÚS FANJUL

DELIVERABLE 2.5– VERSION 1

Nature of the deliverable

R Document, report (excluding the periodic and final reports) X

PU Public, fully open, e.g. web X

More information on the project can be found at http://www.winetwork.eu.

2.1. Data sheet template definition .................................................................................................... 5

2.2. Topic Definition ............................................................................................................................ 6

2.3. Working Procedure. ..................................................................................................................... 7

3.1. Technical data sheets on Good Practices ..................................................................................... 8

3.2. Specific Data Sheets ..................................................................................................................... 9

5.1. APPENDIX 1: GOOD PRUNING PRACTICES .................................................................................. 11

5.2. APPENDIX 2: GLOBAL VINEYARD STRATEGY TO PREVENT GTDs INFECTION .............................. 11

5.3. APPENDIX 3: TRICHODERMA APPLICATION FOR PROTECTING GRAPEVINE PRUNING WOUNDS

5.6. APPENDIX 6: TRUNK RENEWAL .................................................................................................. 11

5.8. APPENDIX 8: PRUNING WITH REGARD TO SAP FLUX ................................................................. 11

5.9. APPENDIX 9: TRUNK CLEANING .................................................................................................. 11

5.10. APPENDIX 10: PEROXYDE HYDROGEN INJECTION .................................................................... 11

5.11. APPENDIX 11: USE OF MYCORRHIZAE IN FIELD........................................................................ 11

5.12. APPENDIX 12: COPPER NANOPARTICULES INJECTIONS ........................................................... 11

5.13. APPENDIX 13: ORANGE OIL APPLICATION ............................................................................... 11

5.14. APPENDIX 14: HOT WATER TREATMENT ................................................................................. 11

5.15. APPENDIX 15: WILD VINES MANAGEMENT ............................................................................. 11

5.16. APPENDIX 16: MATING DISRUPTION........................................................................................ 11

2.1. Data sheet template definition

2.2. Topic Definition

Table 1. Subject of Data Sheet on Good Practices

TOPICS OF TECHNICAL DATA SHEET

Good pruning practices

For the regions where FD is not present yet: Presentation of the

Table 2. TOPICS of Technical Data Sheet on specific region practices

Type of Disease PARTICULAR TECHNICAL DATA SHEET

Pruning with regard sap flux

Copper nanoparticules injections

2.3. Working Procedure.

a) Division of topics in groups :

3.1. Technical data sheets on Good Practices

Table 3 Data Sheet on Good Practices

Languages of Technical Data Sheet

Good pruning Practices

Grapevine Trunk Global vineyard strategy to prevent GTDs infection

Trichoderma application by spraying

Regions where FD is not present yet: Presentation of

Table 4 Data Sheet on specific practices to manage GTD

Languages of Technical Data Sheet

Peroxyde Hydrogen injection

Table 5 Data Sheet on specific practices to FD

Languages of Technical Data Sheet

Hot water treatment

Wild vines management

5.1. APPENDIX 1: GOOD PRUNING PRACTICES

Network for the exchange and transfer of innovative

depending on the training system, corresponding to 15-20%

Botryosphaeria dieback, a GTD caused by numerous

Eutypa dieback, a GTD caused mainly by E. lata, is fre-

Management of pruning Protection of pruning

It is estimated that pruning debris is a potential source

Mechanical grinding and composting on 40 – 50°C

Figure 5: Grapevine pruning system Guyot-Poussard. (http://simonitesirch.com)

Minimal pruning consists of almost no pruning and recently it

Summary - Critical points • Minimize the number of wounds on grapevine in