Professional Documents
Culture Documents
Module 5-Breeding For Disease Resistance
Module 5-Breeding For Disease Resistance
Module 5-Breeding For Disease Resistance
Introduction
Some Concepts of Plant Pathology
a. Bacterial pathogens
b. Fungal pathogens
c. Nematode pathogens
d. Viral pathogens
e. Phytoplasm pathogens
f. Parasitic plants
g. Insect pathogens
III. Genetics of Resistance
IV. Breeding for Resistance to Rusts
V. Breeding for Resistance to Fusarium Head Blight
VI. Breeding for Resistance to Septoria tritici Blotch
VII. Breeding for Resistance to Karnal Bunt
VIII. Breeding for Resistance to Nematodes
-1-
I. Introduction
Early civilizations were well aware that plants were attacked by diseases. There
are references in the Bible to blights, blasts, and mildews (Haggai 2:17, 2 Chronicles
6:28, Amos 4:9). Aristotle wrote about plant diseases in 350 B.C. Theophrastus, the
father of botany (372-287 B.C.), in his Historia Plantarum (History of Plants),
described rusts on grain crops, in meticulous details. During the Middle Ages in Europe,
ergot fungus infected grain and Shakespeare mentions wheat mildew in one of his plays
("He Mildews the White Wheat": King Lear). In India in about the year 500, the
situation arose in which plants were thought to be suffering from human ailments. Thus,
trees and other plants were thought to suffer from wind, bile, phlegm, jaundice and
indigestion. Mathieu Tillet, in 1755, while observing the smuts on wheat, discovered
that there were two types of smut: la carie or what the English called "Common Bunt" or
"Stinking Smut"; and the second type of smut le charbon or what the English called
"loose smut". The distinction between the two types of smut was verified a century later,
in 1847, by Louis and Charles Tulasne. To honor Tillet, they named the "Stinking Smut"
Tilletia caries. In 1807 Isaac Benedict Prevost was the first to provide definitive
evidence that the bunt disease of wheat is caused by a microorganism, smut fungus. He
completed detailed experiments on the germination of bunt spores and demonstrated by
direct inoculation that they could infect wheat. In 1853 Heinrich deBary published a
comprehensive paper that clearly implicated smut and rust fungi as causal organisms of
diseases affecting cereal crops.
In this and the last century, plant breeders have developed cultivars with genetic
resistance to possibly devastating plant pathogens. As this effort has curbed the
widespread famine formerly caused by wheat pathogens it can be regarded among the
most important contributions to agriculture. However, agriculture is a dynamic trade.
Changing agronomic practices and the evolution of new virulent races of pathogens,
requires a persistent and continuous effort in disease management.
An example of this needed vigilance is occurring now (2007) as a new form of
stem rust, has jumped from eastern Africa and is now infecting wheat in Yemen in the
Arabian Peninsula. Researchers with the Global Rust Initiative (GRI) and the
Agricultural Research Service of the United States Department of Agriculture (USDAARS) have confirmed conclusively the existence of the disease in Yemen. There is also
-2-
evidence that the disease has spread into Sudan but more tests are needed to confirm the
finding. Until this discovery, this new strain of stem rust, known as Ug99, had only been
seen in Uganda, Kenya and Ethiopia.
There is precedence for this, from a virulent strain of another wheat disease,
called yellow rust, which emerged in eastern Africa in the late 1980s. Once it appeared in
Yemen, it took just four years to reach wheat fields of South Asia. On its way, this new
strain of yellow rust caused major wheat losses in Egypt, Syria, Turkey, Iran, Iraq,
Afghanistan, and Pakistan, exceeding USD 1 billion in value.
There is every reason to believe the new Ug99 strain of stem rust represents a
much greater risk to world wheat production. Annual losses of as much as $3 billion U.S.
in Africa, the Middle East and south Asia alone are possible.
In order to properly develop a defense against a pathogen, the wheat breeder must know
the attacker. The breeder must understand the pathogens life cycle (inoculation,
infection, proliferation, spread, and latency), its virulence during different
environmental conditions and varying stages of wheat growth, along with its
epidemiology. The breeder must be able to spot signs of the presence of a pathogen and
the breeder must be able to recognize, distinguish, and describe the symptoms that the
wheat plant is displaying. Finally, the breeder must understand the economic impacts of
a particular pathogen in order to determine the amount of resources that should be
directed to resolving the problem.
Inoculation - The introduction of a pathogen to a host.
Infection - The pathogen penetrates and establishes a parasitic relationship with the host.
Proliferation - The reproduction and growth of the pathogen.
Spread - The process by which the pathogen moves within a single host plant or the
process of moving from one infected plant to another.
Latency - A time of inactivity of the pathogen (dormancy in advance of proper
environmental conditions).
Virulence - The degree of pathogenicity of a given pathogen.
Epidemiology - The study of factors influencing the initiation, development, and spread of
infectious disease; the study of disease in populations of plants
-3-
II a) Bacterial pathogens
Bacterial plant pathogens are unicellular microorganisms typically 1 to 3 m in
length. They have neither a well defined nucleus, nor a nuclear membrane. Bacteria are
spread by insects, air currents, splashing rain, and by mechanical means. Free moisture is
usually necessary for infection, and penetration of host tissue occurs through wounds
(created by hail, blowing soil particles, insects, or mechanical injury) or leaf openings
(stomata and hydathodes). Bacteria reproduce chiefly by binary fission, or cell division
yielding two identical daughter cells.
Symptoms caused by bacterial diseases may vary greatly, depending on the
pathogens involved. A bacterial origin is suggested by water-soaked spots or watersoaked margins around lesions consisting of water-congested green tissue in the early
stages of infection. The lesions are greasy and translucent in appearance and may
produce exudates. An exudate consists of droplets of bacterial slime emerging from the
leaf surface through natural openings (stomata, hydathodes).
Fig. 1 Example of a bacterial disease cycle
-4-
II b) Fungal pathogens
Fungal pathogens of wheat are diverse and constitute the most problematic and costly of
the biotic stresses of wheat. Fungi are eukaryotic, non-vascular, saprophytes (taking
nourishment from non-living material), or parasites (removing nutrients in a harmful way
to a living host). Fungi reproduce by means of spores, both asexually and sexually
depending on the species and environmental conditions. Fungal spores spread from plant
to plant through wind, water splashing, and by mechanical means. Fungi infect their host
when a germ tube growing from a spore enters a plant stomata; or by piercing the cuticle
and epidermal cell wall by physical and/or enzymatic means. Within the host, fungi
spread by filamentous growth of hyphae forming a mycelium. The lifecycles of the
different fungal pathogens can be extremely different from one another; and in order to
develop a proper genetic defense against them, a solid understanding of the life cycle of
each pertinent fungal pathogen is important.
Fig. 2 Example of a fungal pathogens life cycle
Further information can be found on a number of fungal pathogens of wheat in the CIMMYT
publications: Bunt and Smut Diseases of Wheat: Concepts and Methods of Disease
Management. Edited by: R. Wilcoxson, and E. Saari; The Septoria Diseases of Wheat:
Concepts and Methods of Disease Management. Edited by: G. Hettel; Septoria and
Stagonospora Diseases of Cereals: A Compilation of Global Research. Edited by: M. van
Ginkel, A. McNab, J. Krupinsky. Available in PDF on line at: http://www.cimmyt.org/ ; Rust
Diseases of Wheat: Concepts and Methods of Disease Management. Edited by: G.P. Hettel.
-5-
II c) Nematode pathogens
Nematodes are a diverse group of worm-like animals. They are found in virtually
every environment, both as parasites and as free-living organisms. They are generally
minute, but some species can reach several meters in length. Plant parasitic nematodes
can be aerial parasites (those feeding on above-ground plant parts) or root and tuber
parasites (those feeding on below ground parts). Nematodes can further be classified as
migratory endoparasites (mobile, feeding inside the plant), sedentary endoparasites (cease
moving once a feeding location inside the plant has been reached), or ectoparasites (feed
on the outside of the plant). The nematode life cycle is typically divided into six stages:
the egg, four juvenile stages, and the adult. The duration of these stages and of the
complete life cycle differs for different species and depends on environmental factors.
Fig. 3 Example of a Typical Nematode Disease Cycle.
feeding on host
cells and tissue
Juvenile 3
Juvenile 4
Second stage
Juveniles hatch
and are attracted
to host root
exudates
Juvenile 2
Adult
Juvenile nematodes
develop inside the
egg
male and female
nematodes mate
and eggs are laid in
the soil
-6-
II d) Viral pathogens
Viruses are sub-microscopic particles consisting of genetic material (either DNA
or RNA) contained within a protein coat known as the capsid. Viruses are obligate
parasites as they can replicate themselves only by the use of another organisms cellular
machinery and intermediate products. Viral infection occurs through wounds, generally
being caused by an insect or nematode vector. Viruses can also be spread by fungi, by
mechanical means or by being carried on in the seed of infected plants.
Fig. 4 Example of Viral Disease Cycle
Spring
Summer
Mites
Infected
winter wheat
Virus
Fall
Mites
Virus
Mites
winter wheat
seedlings
volunteer
wheat
Winter
Virus
Overwintering
winter
wheat
crop and
other
hosts
Arrows indicate
virus movement
Figure adapted from Nebraska Cooperative Extension
-7-
Insect feeds on
annual or perennial
plants, Phytoplasm
is incubating and is
not yet transmitted
Phytoplasm present in
salivary glands in
large numbers and is
injected into plant
while insect is feeding
Phytoplasm spreads by
phloem throughout the
plant
Picture by: S. Mahr; Drawings from: USDA-ARS; Information adapted from: Gianna Sassi Plant Pathology 401 Cornell University
-8-
II f) Parasitic Plants
Parasitic plants are flowering plants that gain some or all of their sustenance from another
plant. They do this by penetrating host plant vascular tissue through modified roots
called haustoria. Striga spp. are obligate root parasites that infect many of the worlds
principal grain and legume crops. Although endemic to Africa, Striga spp. occur in over
40 countries worldwide, ranging from Asia to North America. In sub-Saharan Africa,
more than two-thirds of the 73 million ha of land used for cereal cultivation is severely
infested. Over time, the impact of Striga spp. has increased and it has been described as
one of the most serious biological constraints to food production in Africa (Vasey et al.
2005).
Fig. 6 Example of a
Parasitic Weed Disease
Cycle
Weeks 4-7
Striga emerges
above ground
Week 9-12
Flowering of
Striga
Week 11+
Capsules form
on Striga
Week 11+
Dissemination
of Striga seeds
Host
plant
Aerial Phase
Underground phase
Germination
Days 1-2
Attachment to
host root
Day 2-6
-9-
II g) Insect pathogens
The number of insect pests that feed on wheat is numerous. Insects cause injury
by feeding, and predispose weakened plants to disease. They produce wounds that serve
as infection sites, and often carry pathogens with them from plant to plant. Insects are by
far the most numerous and most efficient vectors of viruses and phytoplasmas. Insects
with chewing mouth parts remove tissue or cut off young plants. Insect larvae feed on,
above and below ground plant parts as well as inhabiting and feeding off of the inside of
the plant. Insects can also be problematic, feeding on the grain both pre- and postharvest.
Fig. 7 Example of an Insect Pathogen Disease Cycle
The pupae overwinter/oversummer
within puparia, the hardened skin of
the last instar larvae. These puaria,
known as the flaxseed stage are
located just below the surface near
the crown of the plant.
Maggots hatch from the eggs in 3 to 7 days, crawl down the leaves,
and feed at the crown or joints along the stem. The maggots develop
through three instars over a 25 to 30 day period.
Source: http://www.oznet.ksu.edu/hessianfly/
- 10 -
- 11 -
Apoplast
Cytoplasm
Host defenses
are suppressed
and disease
occurs
(a)
Signal
transduction
Defenses
activated
(b)
Signal
transduction
Defenses
activated
R
Adapted from McDowell and Woffenden 2003
(c)
Fig. 7 R-Avr Interaction. A pathogen (grey box) has come into contact with a potential host and is
expressing virulence proteins (red). Once inside host cytoplasm, the virulence genes target host defense
response proteins (green). (a) The plant cell does not hold an R-protein capable of recognizing any
virulence protein, and disease results. (b) Pathogen Avr-protein is recognized by a host R-protein,
activating a signal cascade and a defense response. (c) The guard hypothesis: a host defense response
protein has been altered by a virulence protein, this is detected by the host R-protein and a signal
cascade followed by defense mechanisms being activated soon follows.
Plant defense starts with non-self recognition and/or cellular intactness. During
host invasion, pathogens release exogenous as well as endogenous elicitors (Vorwerk et
al. 2006; Huckelhoven 2007). Elicitors are molecules produced by either the host or the
pathogen that in turn induce a response by either the host or the pathogen. Fungal
pathogen elicitors include enzymes which break down the cuticle or cell wall
polysaccharides. Chitin, the major constituent of fungal cell walls is also known to be an
elicitor which triggers a defense response in plants. The bacterial protein, flagellin, is also
an elicitor of host defense. Cuticle and cell wall fragments are examples of endogenous
elicitors of host defense. As these tissues are under physical or enzymatic attack,
released fragments are detected by R-proteins stationed in the cell membrane and a
defense response is initiated.
Within 15 minutes of pathogen recognition, host cells begin producing new
proteins in reaction to the attack (Dangl and Jones 2001). This first response to invasion
is known as a basal defense, and includes production of proteins that inhibit pathogen
enzymes, structural and chemical remodeling of the host cell wall at penetration sites, and
production of antimicrobial agents that will kill the intruder. A second, more extreme,
line of defense is known as the hypersensitive response (HR), or programmed cell
death (PCD) (Greenberg and Yao 2004; Lam et al. 2001; Day and Graham 2007;
- 12 -
Glazebrook 2005). The hyper sensitive response is a strategy of scorched earth where
the plant cell introduces toxic molecules into the surroundings; creating a localized
environment that is incapable of sustaining life for both the pathogen and the cell itself.
The HR is an efficient means of defense against many pathogens, but is ineffective
against necrotrophic fungi.
Breeders have successfully developed lines resistant to diseases by integrating Rgenes into their cultivars for many years; but a durable (sometimes called Horizontal
Resistance, Race non-specific resistance, or Qualitative Resistance), long lasting
resistance in many cases has been difficult to achieve as pathogens quickly evolve and
develop counter resistance genes that circumvent the host cultivars resistance. Breeders
often spot this breakdown in resistance and hurriedly integrate a newly found effective Rgene into their populations. In time, the new R-gene loses its effectiveness and the boombust and induced co-evolution between crop and pathogen continues. An example of this
comes from the United States where in Indiana, 1955; a cultivar was released that held on
R-gene conferring resistance to Hessian fly (Meyetiola destructor) attack. Just six years
later the Hessian fly population had developed a substantial amount of counter-resistance.
By 1964, the Indiana breeders had introduced a second R-gene into their cultivars. This
new R-gene provided resistance for eight years before counter-resistance developed. A
third R-gene was introduced and released in cultivars in 1971; and again was overcome
by the pathogen, this time in a period of ten years (Rausher 2001).
Two methods are available to plant breeders in order to increase the durability of
their resistant cultivars. The first is known as the High-Dose/Refuge or Multiline
strategy (Rausher 2001; Pink 2002). Recall that a hosts resistance to a disease is
conferred by the interaction of its R-gene with the pathogens Avr-gene. A change due to
a mutation of the pathogens allele for an Avr-gene will allow the pathogen to circumvent
or suppress host defenses and cause disease. Returning to the concepts of population
genetics and selection, it is understandable how single R-genes held by the host can be
quickly overcome. Suppose p represents a pathogen populations allele frequency for an
Avr-gene that is recognized by a host plant R-gene, while q represents the allele
frequency of a new recessive allele formed due to mutation:
A1 A1 : P11 = 0.99 p = 0.99 + 0.005 = 0.995
A1 A2 : P12 = 0.01
A2 A2 : P22 = 0.00 q = 0.00 + 0.005 = 0.005
and the selection coefficients for each genotype are:
s11 = 0.90
s12 = 0.70 .
s22 = 0.00
Following the equation for allele frequencies after selection:
q 1 ( ps12 + qs22 )
q1 =
,
1 ( p 2 s11 + 2 pqs12 + q 2 s22 )
one can see that the rare mutant allele becomes predominant in just a small number of
generations:
- 13 -
A multiline or refuge will reduce the selection intensity against the A1A1 and A1A2
genotypes by providing an acceptable host for the pathogen; and maintaining (for an
extended period) a higher frequency of the Avr-gene, recognizable by the wheat cultivars
R-gene, within the pathogen population. Suppose 10 to 20 % of the wheat field contains
susceptible cultivars to the pathogen A1A1 and A1A2 genotypes; the selection intensities
are decreased and the number of generations necessary for the new allele to become
predominant increases dramatically:
The multiline strategy requires that some level of disease is acceptable and that
the pathogen reproduces by sexual means. Also, multilines may not hold a necessary
uniformity that many cropping systems require.
- 14 -
A second option for the plant breeder in generating a cultivar with durable
resistance is know as gene pyramiding. In this strategy several R-genes are deployed in
the same cultivar (McDowell and Woffenden 2003; Pink 2002). In theory, pyramiding
several undefeated R-genes into a single cultivar will provide a more durable resistance
as several mutations would need to take place, one at each of the pathogens
corresponding Avr-loci. With modern molecular biology techniques (See the module:
Biotechnology and Wheat Breeding) it is possible to use markers and probes to track the
introgression of several R-genes into a single cultivar from various sources during a
crossing program.
Although many disease resistances often follows the gene-for-gene model some
follow a race-non-specific, or qualitative mode of resistance (also known as horizontal
resistance) where resistance is controlled by genes with minor to intermediate and
additive effects. Combinations of 3 to 5 of these minor genes can result in a high level of
resistance (Singh et al. 2000).
A third type of resistance is known as partial resistance and most commonly is
associated with slow rusting cultivars. Slow rusting as defined by Caldwell (1968) is a
type of resistance where disease progresses at a retarded rate, resulting in intermediate to
low disease levels against all pathotypes of a pathogen. Partial resistance is a form of
incomplete resistance characterized by a reduced rate of epidemic development despite a
high susceptible infection type. The components that cause slow rusting of a cultivar are
longer latent period, low receptivity or infection frequency, as well as smaller uredial size
and reduced duration and quantity of spore production. All of which can affect disease
progress in the field. Slow rusting resistance has dominated in CIMMYTs bread wheat
improvement program for more than 25 years.
Wheat cultivars susceptible to a disease may in fact show no significant yield
reduction (Zuckerman et al. 2006). Protection of yield, or tolerance, in infected plants
was defined by Caldwell and Shafer (1958) as the ability of a crop to endure severe
epidemics by the pathogen while sustaining only insignificant yield losses as compare
with an infected non-tolerant cultivar. The tolerance that some cultivars demonstrate
over others is not yet well understood, but it has been proposed that storage of
carbohydrates and their mobilization to the sink under stress conditions; or that tolerant
plants have the ability to compensate for the loss of photosynthetic leaf area by increasing
the photosynthetic capacity of the unaffected leaf area contribute to the tolerance to
disease (Zuckerman et al. 1996).
- 15 -
- 16 -
is currently sown to cultivars either derived directly from CIMMYT germplasm or from
CIMMYT germplasm used as parents. For more than 30 years, a major proportion of the
CIMMYT wheat germplasm and germplasm developed by other breeding programs have
remained resistant to stem rust. Resistance gene Sr31, located on rye translocation 1B.1R
contributed to high level of resistance in several wheat cultivars developed worldwide in
recent years. Consequently stem rust disease is often not considered important and in
many countries wheat breeding is currently done in the absence of stem rust and research
in this area in the last two decades also declined substantially.
Detection in 1999 of Pgt race Ug99 in Uganda with broad virulence, including the
virulence for Sr31, and its migration to Kenya and Ethiopia has been recognized as a
highly significant event and led to the launch of Global Rust Initiative during 2005. All
major cultivars currently grown in North Africa, the Middle East and Asia are moderately
or highly susceptible to this race. Predominant wind patterns or human errors are likely
to introduce this race to above regions and beyond. One of the major challenge to wheat
breeding is identifying or developing and diffusing adapted resistant cultivars before this
migration occurs. Although some race-specific resistance genes, mostly of alien origin,
viz., Sr22, 24, 25, 26, 27, 29, 32, 33, 35, 36, 39, 40, 44, R (1A.1R translocation) and
Tmp, can provide effective control; not all can be used in developing cultivars as some of
these alien translocations are associated with negative effects on grain yield or quality.
Shortening of these alien segments could help their utilization. Improved wheat
germplasm that carry Sr24, Sr25, Sr26, SrTmp and SrR genes is already identified and
can be used in breeding. Gene Sr24 currently occurs with a relatively high frequency in
wheat cultivars and has become ineffective in India and South Africa soon after the
release of cultivars that carried this resistance gene. The best strategy therefore is to
reconstitute durable adult-plant resistance that once protected the green revolution and
subsequent wheat cultivars. If race-specific genes need to be used, they must be
deployed in combination to enhance their longevity.
Durable stem-rust resistance of some older US, Australian and CIMMYT spring
wheats is believed to be due to the deployment of Sr2 in conjunction with other unknown
minor, additive genes. McFadden transferred Gene Sr2 to hexaploid wheat in the 1920s
from tetraploid emmer wheat cultivar Yaroslav. The slow rusting gene Sr2 confers by
itself only moderate levels of resistance. Its presence can be detected through its complete
linkage with the pseudo-black chaff phenotype. A large number of wheat lines were
evaluated during 2005 in Kenya under the stem rust epidemic caused by Ug99.
Genotypes with pseudo-black chaff phenotype showed varying degrees of disease
severity with a maximum severity reaching to about 60% compared with 100% severity
for highly susceptible materials. Moreover, the host reaction for these genotypes on the
same internode varied from moderately resistant to susceptible. These observations
clearly indicated that although slow rusting resistance gene Sr2 continues to confer at
least some resistance, the level of resistance was not sufficient when this gene is present
alone under high disease pressure in Kenya. Sr2 was detected in several highly resistant
old, tall Kenyan cultivars, including Kenya Plume (Singh and McIntosh 1986), and
CIMMYT-derived semidwarf cultivar Pavon 76. These cultivars have shown a
maximum disease score of 15MR (15% disease severity with moderately resistant
reaction). Because Pavon 76 is susceptible to race Ug99 at the seedling stage, its
resistance, as speculated earlier (Rajaram et al. 1988), is based on multiple additive genes
- 17 -
where Sr2 is an important component. Wide testing of improved wheat germplasm also
has helped in identifying additional sources of adult-plant resistance. These sources are
being used at CIMMYT to incorporate durable stem-rust resistance into high yielding,
widely adapted wheat cultivars using the methodology described below.
Breeding for durable resistance based on minor additive genes has been
challenging and often slow, for several reasons: 1) a sufficient number of minor genes
may not be present in a single source genotype, 2) a source genotype may be poorly
adapted, 3) there may be confounding effects from the segregation of both major and
minor genes in the population, 4) crossing and selection schemes and population sizes are
more suitable for selecting major genes, 5) reliable molecular markers for several minor
genes are unavailable, and 6) the cost associated with identifying and utilizing multiple
markers is high. One suggested approach is to use recurrent selection schemes to
accumulate several minor genes in a single genetic background. Such selection schemes
have often been more of a scientific interest than actually being applied in breeding.
Selection for resistance alone will not generate important popular cultivars, unless it is
simultaneously combined with other traits, such as high yield and quality. However, such
germplasm carrying combinations of minor genes should be very useful in transferring
these genes to modern cultivars.
A successful example of breeding for resistance based on minor genes is the
resistance to leaf and stripe rusts in wheat, which took about 30 years of continuous effort
at CIMMYT. In the early 1970s, S. Rajaram, influenced by the concept of slow-rusting
resistance in wheat proposed by R. Caldwell and partial resistance to late blight of potato
put forth by J. Niederhauser, made a strategic decision: to initiate selection for slowrusting resistance to leaf rust in CIMMYT spring wheat germplasm. In the early phase of
breeding he maintained plants and lines in segregating populations that would show 2030% rust severity with compatible infection type. This strategy led to the release of
several successful wheat cultivars, such as Pavon 76 and Nacozari 76, in Mexico and
other countries. These slow-rusting lines were used heavily in the crossing program and
resulted in the wide distribution of minor genes within CIMMYT spring wheat
germplasm.
The genetic basis of such resistance started to become clear in the early 1990s.
High-yielding lines that combine four or five additive, minor genes for both leaf and
stripe rusts and show near-immune levels of resistance were developed in the 1990s
(Singh et al. 2000). Three or four lines carrying different minor genes were crossed (3way and 4-way crosses), and plants in large segregating populations were selected under
artificially created rust epidemics. Races of pathogens that have virulence for racespecific resistance genes present in the parents were used to create the epidemics. The
resulting highly resistant lines are now being used in a planned manner to transfer these
minor resistance genes to well adapted, farmers choice cultivars that are currently
grown across large areas but have become susceptible to rust races in Mexico. Based on
genetic information on the number of additive, minor genes that must be transferred to
achieve the desired level of resistance, the crossing and selection scheme described below
was developed and applied. This strategy has allowed simultaneous transfer not only of
resistance genes but also other minor genes with small effects that increase the yield
potential or improve the grain quality of an adapted cultivar.
- 18 -
- 19 -
- 20 -
In 2006 CIMMYT modified their FHB screening system for greater screening
capabilities, accuracy and precision. These changes have included the following:
Changing the primary screening site from Toluca to El Batan (headquarters),
Mexico
Implementing an automated, programmable misting system
Using precision CO2 sprayers for liquid inoculum application
Figure 11: CIMMYT Primary FHB Screening Site, El Batan, Mexico: A) Micro-sprinkler of misting
system, B) CO2 backpack sprayer used for inoculations C) Examples of symptoms observed.
Fine-Misting System
Approximately 1.9 hectares were placed under a programmable misting system to
provide uniform humidity conditions for favorable FHB conditions. This misting system
includes about 1,600 DAN modular micro-sprinklers (Figure 1A) spaced at distances of 3
x 4 meters, and operated automatically by a programmable timer to provide humid
conditions 24 hours a day. The system is programmed for misting both day and night,
with misting intervals and duration varying depending on the time of day (more frequent
misting during the driest periods of the day).
CO2 Sprayers
CO2 backpack sprayers were purchased from R&D Sprayers with an 8002VS flat
fan nozzle for precise application of inoculum (Figure 1B). Liquid inoculum was applied
at a concentration of 100,000 macroconidia/ml of a mixture of three isolates of F.
graminearum at a pressure of 40psi and a rate of 39ml of inoculum per meter. Wheat
plots were first sprayed at anthesis, and barley plots were first sprayed at heading.
Inoculum was re-applied at the same rate three days after the first inoculation. Grain
spawn inoculum (colonized maize grains) was also spread in the FHB evaluation plots to
ensure adequate levels of the pathogen for the generation of the disease.
By screening germplasm using the method above, recommendations were made to
CIMMYT breeders on germplasm for further use of these materials by their programs
and development of new FHB resistant wheat (Table 1).
- 22 -
Table 1 Accessions That Have Been Recommended for Further Use Based on
Recent (2006) FHB Trials in a Disease Nursery at CIMMYT
Cross
Selection History
SHANGHAI
-33GH-0M-0Y-0Y-0SCM-0FGR-0FGR-0FGR
GONDO/TNMU
CMSS92M01425S-015M-0Y-0Y-050M-16Y-2M-0Y-2SJ-0Y 0FGR0FGR
IVAN/6/SABUF/5/BCN/4/RABI//GS/CR
A/3/AE.SQUARROSA (190)
CMSS00M00007S-030M-1Y-4SCM-010Y-0FGR
IVAN/6/SABUF/5/BCN/4/RABI//GS/CR
A/3/AE.SQUARROSA (190)
CMSS00M00007S-030M-1Y-8SCM-010Y-0FGR
IVAN/6/SABUF/5/BCN/4/RABI//GS/CR
A/3/AE.SQUARROSA (190)
CMSS00M00007S-030M-1Y-24SCM-2Y-0FGR
GONDO/FINSI
CMSS00Y02909S-030Y-030M-030Y-2M-1M-0Y
TNMU/6/CEP80111/CEP81165/5/MRN
G/4/YKT406/3/AG/ASN//ATR
CMBW91Y01692S-13Y-2AL-3AL-010Y-3M-0Y-3PZ-0Y
MAYOOR//TK SN1081/AE.SQUARROSA
(222)/4/CS/LE.RA//CS/3/PVN/5/PRINIA
CSSS00B00011S-2Y-10M-2Y-8FGR-1Y-0FGR-0BI
80456/YANGMAI 5//SHA5/WEAVER/3/PRINIA
CMSS98M00896T-040Y-0100M-040Y-040M-030Y-53M-1Y-0M
SUM3/3/CS/LE.RA//CS/4/YANGMAI 158
-0FGR
WUH1/VEE#5//CBRD
CMSS92M01863S-015M-0Y-050M-0Y-13M-0Y-0FGR-0FGR-0FGR
CMSS97M01288S-030M-020Y-030M-015Y-29M-2Y-2M-0Y020SCM
CMSS97M01295S-040M-020Y-030M-015Y-40M-1Y-2M-0Y020SCM
CMSS97M01318S-040M-20Y-010M-010Y-5M-2Y-2M-0Y-020S
CMSS97M01333S-030M-100Y-010M-010Y-6M-1Y-2M-0Y020SCM
CMSS98M00761T-040Y-0100M-040Y-040M-030Y-15M-2Y-0M
80456/YANGMAI 5//SHA5/WEAVER
NG8675/CBRD//SHA5/WEAVER
GONDO/CBRD
BAU/MILAN//CBRD
EMB16/CBRD//CBRD
80456/YANGMAI
5/3/PF70354/BOW//DUCULA/4/DULUS
CMSS99Y03242T-040M-040Y-040M-030Y-030M-17Y-2M
YANGMAI 5*2/4/MOR/VEE#5//DUCULA/3/DUCULA
CMSS99Y03149F-040M-040Y-040M-030Y-030M-15Y-2M-2M-0Y
SHA4/CHIL/4/CAR422/ANA//TRAP#1/3/STAR
CMSS99M02404S-040M-030Y-030M-2Y-1M-2M-0Y
SHA3/SERI//G.C.W1/SERI/3/SHA3/SERI//YANG87142
CMSS00Y02958S-030Y-030M-030Y-5M-1M-0Y
NG8675/CBRD//MILAN/3/NG8675/CBRD
CMSS95M01814T-050Y-0100M-050Y-050M-040Y-030M-3Y-0M0SCM-0Y-0FGR-0FGR
-0CHN-0SCM-0Y-0SCM-0Y-0FGR-0FGR
TINAMOU
CM81812-12Y-06PZ-5Y-5M-0Y-3AL-0Y-2M-010Y-0M-3PZ-0Y
TRAP#1/BOW//TAIGU
DERIVATIVE
CMSS94Y01180S-0300M-0100Y-050Y-050M-41Y-030M-3PZ0Y-3M-0Y-0SCM-0Y-0FGR-0FGR
TRAP#1/BOW//TAIGU
DERIVATIVE
CMSS94Y01180S-0300M-0100Y-050Y-050M-41Y-030M-1SJ0Y-2M-0Y-0SCM-0Y-0FGR-0FGR
SHA3/CBRD
-0SHG-1GH-0FGR-0FGR-0SCM-0FGR-0FGR-0FGR
SUM3/3/CS/LE.RA//CS/4/YANGMAI 158
-0FGR-0FGR-0FGR
YANGMAI 5
-0CHN-0SCM-0Y-0SCM-0FGR-0FGR-0FGR
TRAP#1/BOW//TAIGUDERIVATIVE
CMSS94Y01180S-0300M-0100Y-050Y-050M-41Y-030M-3PZ0Y-0FGR-0FGR
EMB27/KLORI
F37023-901F-904F-902F-903F-901F-453F-901F-900F
BR23/EMB27
-7TSB-0Y-0SCM-0Y-0SCM-0Y-0FGR-0FGR
- 23 -
- 24 -
Wheat seedlings are evenly sprayed with spore suspension to run off.
Leaves are left to dry for 30 min.
Three 5 cm sections cut from the middle of the primary leaves.
Water agar is prepared (10g/L) containing 100mg/L benzimidazole (used to
retard senescence)
50 mL dispensed into an appropriate number of 8x12x2 cm clear polystyrene
boxes.
Rectangular sections (3x9 cm) are cut from the center of the agar.
8 to 10 Seedling leaf sections per box are laid, top surface up, across the
gap(created by cutting out of the rectangular sections) so that the cut ends rest
on the agar.
Strips of agar are laid over the cut edges of the leaves.
The boxes are closed and covered with black plastic or foil.
The boxes are incubated at 20C for 48 h in darkness.
The boxes, then, are uncovered and left under white phosphorescent light for
12 days at 20C.
Boxes are then moved to conditions with near-ultraviolet light at 15C to
promote sporulation (14 days after inoculation)
The percentage of leaf area covered by lesions bearing pycnidia is scored 4 to
5 times at intervals of 2-4 days for 19-28 days after inoculation. (Assessments
carried out under a 40x dissecting microscope).
New computer tools may lend themselves useful in scoring of diseases. Image analysis
software can be a faster and more reliable method for scoring of diseases than that which
can be done with the eye which is more subjective and can tire. For further information
on how image analysis software can help quantify disease incidence see Mirik et al.
(2006).
In field studies at CIMMYT disease is rated using a double digit scale:
After anthesis, spot blotch severity is evaluated using the double-digit scale (00-99)
developed as a modification of Saari and Prescotts scale for assessing severity of foliar
diseases of wheat (Saari and Prescott, 1975; Eyal at al., 1987). The first digit (D1)
indicates disease progress in canopy height from the ground level, while the second digit
(D2) refers to severity measured based on diseased leaf area. Both D1 and D2 are scored
on a scale of 1 to 9. Since the rate of disease progress in the field can be extremely high
in some regions, it is often needed to take repeated scores to properly assess the level of
resistance (Dubin et al., 1998; Duveiller et al., 1998). It is recommended to take several
individual disease scores per plot at 3- to 7-day intervals over a 3- to 4-week period
between anthesis and dough stage depending on seeding date. For each score, percent
disease severity is estimated based on the following formula:
% severity = ((D1/9) x (D2/9) x 100)
The area under the disease progress curve (AUDPC) is calculated using the
percent severity estimates corresponding to the three to four ratings shown below:
,
- 25 -
where, xi = severity on the ith date, ti = ith day, and n = number of dates on which disease
is recorded. The AUDPC (%-day) measures, the amount of disease as well as the rate of
progress.
It may be appropriate to standardize the AUDPC to take into account different
epidemics situations and planting dates. In this case AUDPC should be divided by the
total number of days in the evaluation period (AUDPC/day) to better compare genotypes
or among epidemics.
A good example of how to identify sources of race-specific resistance is
underway at the CIMMYT station in Toluca. Ten nurseries, each containing the same 30
wheat cultivars replicated twice in each nursery have been planted. Each nursery has been
inoculated with a different strain of Mycosphaerella graminicola, and disease has been
scored as described above four times over the season. By doing this, the breeder can
identify which cultivars hold R-genes for specific pathogen isolates. Cultivars holding
resistance to different races of the pathogen can then be put into a double cross scheme in
an attempt to introgress a number of R-genes into a single cultivar.
A number of cultivars that can be used as a source for resistance are listed in table
2 taken from a study done by Chartrain et al. (2004).
- 26 -
Table 2 Sources of Resistance to STB and Percentage of Leaf Area Covered by Lesions Bearing Pycnidia of 12 Different
Mycosphaerella graminicola isolates
Wheat line
Origin of line
CA30
USA
IPO001
NLANDS
IPO323
NLANDS
IPO87019
URUGUAY
IPO88004
ETHIOPIA
IPO89011
NLANDS
IPO90004
MEXICO
IPO90012
MEXICO
IPO92006
PORTUGAL
IPO94269
NLANDS
ISR398
ISRAEL
ISR8036
ISRAEL
SENAT
GENE
TE9111
MILAN
ISRAEL493
CHAUCER
EQUINOX
ARINA
TONIC
MENTANA
FLAME
VERNOPOLIS
REAPER
OLAF
RIBAND
LONGBOW
BULGARIA88
FRONTANA
CATBIRD
BALDUS
SHAFIR
CHINESE
SPRING
KK (L6.A.4)
COURTOT
Isolate mean
Denmark
USA
Potugal
CIMMYT
Israel
UK
UK
Switzerland
UK
Italy
UK
Brazil
UK
USA
UK
UK
Bulgaria
Brazil
CIMMYT
Netherlands
Israel
1
6
17
9
6
18
3
45
25
20
25
7
16
29
49
23
21
35
3/
71
18
10
8
10
48
24
7
18
37
24
29
59
3
45
44
34
50
38
4
51
64
0
0
0
0
7
71
65
0
63
39
3
0
64
1
71
57
5
53
1
30
2
1
20
0
10
21
1
17
37
31
49
37
9
22
30
34
27
51
39
3
44
66
0
53
4
77
0
0
22
7
8
42
30
0
54
3
0
0
44
71
13
2
95
6
1
33
1
12
23
36
8
6
5
59
58
41
47
38
38
37
16
43
63
55
17
20
50
12
7
27
24
8
23
38
33
3
36
0
30
42
36
37
81
54
58
7
37
1
19
6
6
28
82
84
47
66
3
48
1
56
72
57
67
89
78
87
3
39
17
25
44
5
61
78
59
44
64
53
69
52
59
54
65
69
80
66
59
25
0
12
9
16
30
30
44
24
3
27
17
34
15
32
35
28
5
55
55
24
6
10
0
34
6
30
28
19
14
29
15
18
41
18
24
42
18
35
0
26
44
2
0
1
29
30
2
10
6
9
41
2
29
9
24
13
24
37
17
49
9
46
China
38
44
15
96
55
90
99
57
42
16
39
CIMMYT
France
68
59
20
10
88
33
1
81
55
0
34
26
78
76
44
35
4
26
78
89
39
93
100
64
79
79
53
13
63
29
0
66
24
4
73
14
Grey shaded figures indicate significance for specific interactions of wheat line with M. graminicola isolates Wheat lines in bold are those with best resistance.
- 27 -
It has been reported that selection for resistance to Karnal bunt is exceptionally difficult
as environmental conditions play a significant role in this pathogens virulence and
nurseries are both expensive and difficult to maintain (Datta et al. 1999; SukhwinderSingh et al. 2003).
VIII. Nematodes
It has been estimated that plant-parasitic nematodes account for 10% of
worldwide crop loss. Cereal Cyst Nematodes are a complex group of 12 described and
several undescribed species with Heterodera avenae being the most economically
important. Yield losses from H. avenae vary from 15-20% in Pakistan to 40-92% in
Saudi Arabia. The damage threshold is dependent on a number of variables including
wheat cultivar, soil type, the pathotype and ecotype of the nematode, and climactic
conditions of the geographical region. Potential damage from nematodes is increased
where wheat is grown in stressful environments such as poor soil nutrition, temperature
stress, water stress, or where there is pressure from other pathogens (Nicol et al. 2003). .
Identification of Cereal Cyst and Lesion nematodes is difficult, and has
historically been accomplished through comparative morphology. A key to the
identification of nematodes can be found at: http://nematode.unl.edu/nemakey.htm and
may be useful in identifying a specific nematode pest. More recently laboratory
techniques including AFLP markers have been developed to identify and discriminate
both cyst and lesion nematodes (Andrs et al. 2001; Mokabli et al. 2001; Orui and
Mizukubo 1999; Curran 2002).
A handful of Cyst resistance genes (Cre) have been identified in wheat and have
been deployed to provide effective resistance in parts of the world. In Europe and North
Africa, Cre1, provides good resistance against H. avenae; but has proven ineffective
against those races specific to Asiatic and Australian wheat growing regions. Greater
regional usability is offered by Cre2 and Cre4 which come from Aegilops spp. Table 2
contains a number of sources that could be used to introgress nematode resistance genes
into a breeding population. Host resistance to nematodes is still poorly understood and
collaboration between research institutions like CIMMYT and country programs will be
necessary to improve and develop more reliable host resistance.
Table 3 suggests that both horizontal and vertical resistance to nematodes is
available in existing germplasm. If nematode resistance is a part of a programs breeding
objectives, cultivars with proven resistance should be chosen as a donor parent in a
backcross strategy with a locally adapted recurrent parent. Stated earlier, nematode
pressure depends on many variables and successfully screening and selecting for
cultivars holding resistance genes may be difficult as nematode pressure will vary from
site to site and from year to year. Establishing a disease nursery for nematode resistance
selection would be helpful. In order to keep nematode populations at a maximum an
understanding of the problem nematodes preferred environment is necessary. By
maintaining an optimal soil ecosystem for the nematodes, and offering them susceptible
hosts (also useful as checks) season after season in the same location; a large population
of nematodes will be maintained, offering better disease pressure and an opportunity for
more effective selection.
- 28 -
Table 3. Principal sources of genes used for breeding resistance to Heterodera avenae in cerealsb
Cereal species
Cultivar or line
Origin
Remarksc
Genetic information
Used
Wheat
Triticum aestivum
-, Australia
Katyil
Australia
Ccn1
S, India
Australia
pR in cv Molineux
Australia
Festiguay
AUS4930
48'
T. durum
Australia
=
'Iraq Iraq
Psathias
S, to some pathotypes
also pR
7654,
Sansome,
7655, -
S, to some pathotypes
Khapli
France
also pR
CPI 110813
Central Asia
Ae. tauschii
(T. tauschii)
AUS 18913
1 dominant gene on chromosome 2DL, R, Australia (Ha13) and several other Australia advanced breeding lines
Cre3
countries
T. variabilie
West Asia
Gene Rkn-mn1 on chromosome 3U or R, to various pathotypes and France, Algeria, Spain, India, Syria
Meloidogyne naasi and H. latipons
3Sv
T. longissimum
18
T. ovatum
79
Mediterranean
basin
T. triunciale
(Ae. triuncialis)
TR-353
Spain
1 dominant
CreAet)
T. geniculata
(Ae. geniculata)
Spain, Bulgaria,
T. ventricosum
(Ae. ventricosa)
VPM 1
Jordan, Tunisia
On chromosome 2AS, Cre5 (formerly populations and H. latipons R, to France, Australia (under evaluation)
CreX)
French pathotype (Ha12)
T. ventricosum
(Ae. ventricosa)
Mediterranean
basin
T. ventricosum
(Ae. ventricosa)
gene,
Cre7
R, Australia (Ha 13) and several other Australia synthetic hexaploid lines
countries
1 dominant gene on chromosome 5Nv, R, to Australian pathotype (Ha13), not Spain, Australia (under evaluation)
Cre6
effective against Spanish (Ha71)
- 29 -
- 30 -
References:
Anderson, J.A., R.W. Stack, S. Liu, B.L. Waldron, A.D. Fjeld, C. Coyne, B. Moreno-Sevilla, F.J. Mitchell,
Q.J. Song, P.B. Cregan, and R.C. Frohberg. 2001. DNA markers for Fusarium head blight
resistance QTLs in two wheat populations. Theor. Appl. Genet. 102: 1164-1168.
Andrs M.F., M.D. Romero, M.J. Montes, and A. Delibes. 2001. Genetic relationships and isozyme
variability in the Heterodera avenae complex determined by isoelectric focusing. Plant Pathology
50: 270-279.
Arraiano, L.S., and J.K.M. Brown. 2006. Identification of isolate-specific and partial resistance to
Septoria tritici blotch in 238 European wheat cultvars and breeding lines. Plant Pathology 55: 726738.
Buerstmayr, H., M. Lemmens, L. Hartl, L. Doldi, B. Steiner, M. Stierschneider, and P. Ruckenbauer. 2002.
Molecular mapping of QTLs for Fusarium head blight resistance in spring wheat. I. Resistance to
fungal spread (type II resistance). Theor. Appl. Genet. 104: 84-91.
Caldwell, R.M. 1968. Breeding for General and/or Specific Plant Disease Resistance. Proc. 3rd Int.
Wheat Genetics Symp Canberra, Australia. Pp. 263-272..
Caldwell, R.M., and J.E. Shafer. 1958. Tolerance to cereal leaf rust. Science 128: 714-715.
Chartrain, L., P.A. Brading, J.C. Makepeace, and J.K.M. Brown. 2004. Sources of resistance to Septoria
tritici blotch and implications for wheat breeding. Plant Pathology 53: 454-460.
Chartrain, L., S.T. Berry, and J.K.M. Brown. 2005. Resistance of line Kavkav.K400 L.6.A.4 to Septoria
tritici blotch controlled by isolate-specific resistance genes. Phytopathology 95: 664-671.
Curran, J. 2002. Quantifying nematodes and other root diseases in the Australian wheat belt. Fourth
International Congress of Nematology, 8-13th June 2002, Tenerife, Spain. Nematology 4(2), 126.
[Abstr.]
Dangl, J.L., and J.D.G. Jones. 2001. Plant Pathogens and Integrated Defense Responses to Infection.
Nature 411:826-833.
Datta, R., Singh Harjit, V.S. Gupta, P.K. Ranjekar, and H.S. Dhaliwal. 1999. Gene-for-gene relationship
for resistance in wheat to isolates of Karnal bunt, Neovossia indica. Plant Breeding 118: 362-364.
Day, B., T. Graham. 2007. The Plant Host-Pathogen Interface: Cell Wall and Membrane Dynamics of
Pathogen Induced Responses. Annals of the New York Academy of Sciences. On line
doi:10.1196/annals.1391.029, from the Annals volume Stress Responses in Biology and Medicine.
Dubin, H.J., Arun, B., Begum, S.N., Bhatta, M, Dhari, R., Goel, L.B., Joshi, A.K., Khanna, B.M., Malaker,
P.K., Pokhrel, D.R., Rahman, M.M., Saha, N.K., Shaheed, M.A., Sharma, R.C., Singh, A.K., Singh,
R.M., Singh, R.V., Vergas, M., Verma, P.C., 1998. Result of South Asian regional
Helminthosporium leaf blight and yield experiment 1993-1994. In: Duveiller, E., Dubin, H.J.,
Reeves, J., McNab, A. (Eds.), Helminthosporium blights of wheat: Spot blotch and tan spot.
CIMMYT, Mexico , D.F., pp. 182-187.
Duveiller, E., C. Bragard and H. Maraite. 1997. Bacterial Leaf Streak and Black Chaff Caused by
Xanthomonas translucens. In E. Duveiller, L. Fucikovsky, and K. Rudolph (eds.), The Bacterial
Diseases of Wheat: Concepts and Methods of Disease Management, D.F.: CIMMYT.
Duveiller, E., Garca, I., Toledo , J., Franco, J., Crossa, J. and Lopez, F. 1998. Evaluating spot blotch
resistance of wheat: improving disease assessment under controlled conditions and in the field.
Proceedings of the International Workshop on Helminthosporium Diseases of Wheat: Spot Blotch
and Tan Spot, CIMMYT, El Batn, Feb. 9-14, 1997, Mexico , 171-181.
Duveiller, E., R.P. Singh, and J.M. Nicol. 2007. The challenges of maintaining wheat productivity: pests,
diseases, and potential epidemics. Euphytica 157: 417-430.
Eyal, Z., A.L. Scharen, J.M. Prescott, and M. van Ginkel. 1987. The Septoria disease of wheat: concepts
and methods of disease management. Mexico , D.F.: CIMMYT.
Fuentes- Davila, G., and S. Rajaram. 1994. Sources of resistance to Tilletia indica in wheat (Triticum
aestivum). Crop Protect 13:2024.
Fuentes-Davila, G., S. Rajaram, and G. Singh. 1995. Inheritance of resistance to Karnal Bunt (Tilletia
indica Mitra) in bread wheat (Triticum aestivum L.). Plant Breeding 114: 250-252.
- 31 -
- 32 -
Singh, R.P. 1992b. Genetic association of leaf rust resistance gene Lr34 with adult plant resistance to stripe
rust in bread wheat. Phytopathology 82: 835-838.
Singh, R.P. and S. Rajaram. 1994. Genetics of adult plant resistance to stripe rust in ten spring bread
wheats. Euphytica 72: 1-7.
Singh, R.P. and S. Rajaram. 2002. Breeding for disease resistance in wheat. In B.C. Curtis, S. Rajaram
and H. Gomez Macpherson (eds.), Bread Wheat Improvement and Production. FAO, Rome.
Singh, R.P. and S. Rajaram. 1992. Genetics of adult-plant resistance to leaf rust in Frontana and three
CIMMYT wheats. Genome 35: 24-31.
Singh, R.P., A. Kazi-Mujeeb, and J. Huerta-Espino. 1998. Lr46: a gene conferring slow rusting resistance
to leaf rust in wheat. Phytopathology. 88: 890-894.
Singh, R.P., and R.A. McIntosh. 1986. Genetics of resistance to Puccinia graminis tritici and Puccinia
recondita tritici in Kenya plume wheat. Euphytica 35: 245-256.
Singh, R.P., H. Ma, and S. Rajaram. 1995. Genetic analysis of resistance to scab in spring wheat cultivar
Frontana. Plant Dis. 79: 238-240.
Stinger, L.C., C. Twyman, and D.S.G. Thomas. 2007. Learning to reduce degradation on Swazilands
arable land: Enhancing understanding of Striga asiatica. Land Degrad. Develop. 18: 163-177.
Sukhwinder-Singh, G.L. Brown-Guedira, H.S. Dhaliwal, J.C. Nelson, and H. Singh. 2003. Mapping of a
resistance gene effective against Karnal bunt pathogen of wheat. Theoretical and Applied Genetics
106: 287-292.
Van Ginkel, M., I. Ortiz-Monasterio, R.M. Trethowan, and E. Hernandez. 2001. Methodology for
selecting segregating populations for improved N-use efficiency in bread wheat. Euphytica 119: 223
230.
Vasey, R.A., J.D. Scholes, and M.C. Press. 2005 Wheat (Triticum aestivum) Is Susceptible to the Parasitic
Angiosperm Striga hermonthica, a Major Cereal Pathogen in Africa. Phytopathology 95: 12941300.
Villareal, R.L., A. Mujeeb-Kazi, G. Fuentes- Davila, and S. Rajaram. 1996 Registration of four synthetic
hexaploids germplasm lines derived from Triticum turgidum T. tauschii crosses and resistant to
Karnal bunt. Crop Sci 36: 218.
Vorwerk, S., S. Sommerville, and C. Somerville. 2004 The Role of Plant Cell Wall Polysaccharide
Composition in Disease Resistance. Trends in Plant Science 9: 203-209.
William, M., R.P. Singh, J. Huerta-Espino, S. Ortiz Islas, and D. Hoisington. 2003b. Molecular marker
mapping of leaf rust resistance gene Lr46 and its association with stripe rust resistance gene Yr29 in
wheat. Phytopathology 93: 153-159.
Zadoks, J.C. 2003. Two wheat Septoria; two emerging diseases from the past. In: G. Kema, M. van
Ginkel, M. Harrabi (eds.), Global insights into the Septoria and stagonospora diseases of cereals.
Proceedings of the 6th international symposium on Septoria and stagonospora diseases of cereals,
Dec 8-18, 2003, Tunis, Tunisia, Pp. 1-12.
Zuckerman, E., A. Eschel, and Z. Eyal. 1996. Physiological aspects related to tolerance of spring wheat
cultivars to Septoria tritici blotch. Phytopathology 87: 60-65.
- 33 -