Journal of Economic Entomology, 116(2), 2023, 275–288

https://doi.org/10.1093/jee/toac154
Advance Access Publication Date: 4 January 2023
Research

Special Collection: Global Perspectives on Field-Evolved Resistance to

Transgenic Bt Crops

Monitoring Insect Resistance to Bt Maize in the European

Union: Update, Challenges, and Future Prospects
Matías García,* Carlos García-Benítez,* Félix Ortego, and Gema P. Farinós1,

Downloaded from https://academic.oup.com/jee/article/116/2/275/6968981 by guest on 30 April 2023

Laboratory of Applied Entomology for Human and Plant Health, Centro de Investigaciones Biológicas Margarita Salas, CSIC, 28040
Madrid, Spain and 1Corresponding author, e-mail: gpfarinos@cib.csic.es
*Both authors contributed equally to this work.

Subject Editor: Yidong Wu

Received 6 July 2022; Editorial decision 8 September 2022.

Abstract
Transgenic maize producing the Cry1Ab toxin of Bacillus thuringiensis (Bt maize) was approved for cultivation
in the European Union (EU) in 1998 to control the corn borers Sesamia nonagrioides (Lefèbvre) and Ostrinia
nubilalis (Hübner). In the EU since then, Cry1Ab is the only Bt toxin produced by Bt maize and Spain is the only
country where Bt maize has been planted every year. In 2021, about 100,000 hectares of Bt maize producing
Cry1Ab were cultivated in the EU, with Spain accounting for 96% and Portugal 4% of this area. In both coun-
tries, Bt maize represented less than 25% of all maize planted in 2021, with a maximum regional adoption of
64% Bt maize in northeastern Spain. Insect resistance management based on the high-dose/refuge strategy
has been implemented in the EU since 1998. This has been accompanied by monitoring to enable early detec-
tion of resistance. The monitoring data from laboratory bioassays show no decrease in susceptibility to Cry1Ab
had occurred in either pest as of 2021. Also, control failures have not been reported, confirming that Bt maize
producing Cry1Ab remains effective against both pests. Conditions in the EU preventing approval of new ge-
netically modified crops, including maize producing two or more Bt toxins targeting corn borers, may limit the
future effectiveness of resistance management strategies.

Resumen
El maíz transgénico que expresa la toxina Cry1Ab de Bacillus thuringiensis (maíz Bt) fue aprobado para su
cultivo en la Unión Europea (UE) en 1998 para el control de los taladros del maíz Sesamia nonagrioides
(Lefèbvre) y Ostrinia nubilalis (Hübner). Desde entonces en la UE sólo se ha sembrado maíz Bt que produce
la toxina Cry1Ab, y España es el único país donde se cultiva ininterrumpidamente desde 1998. En 2021 se
cultivaron en la UE unas 100.000 hectáreas de maíz Bt que produce Cry1Ab, de las cuales el 96% corresponde
a España y el 4% a Portugal. En ambos países, el maíz Bt representó menos del 25% de todo el maíz sembrado
en 2021, con una adopción regional máxima del 64% de maíz Bt en el noreste de España. El manejo de la
resistencia de insectos, basado en la estrategia dosis-alta/refugio se ha aplicado en la UE desde 1998. Esto
ha ido acompañado de un seguimiento que permite la detección temprana de la resistencia. Los resultados
del seguimiento a partir de bioensayos de laboratorio muestran que hasta 2021 no se ha producido una
disminución de la susceptibilidad a Cry1Ab en ninguna de estas plagas. Además no se han registrado fallos en
el control, confirmándose que el maíz Bt que produce Cry1Ab sigue siendo eficaz frente a ambas plagas. Las
condiciones de la UE que impiden la aprobación de nuevos cultivos genéticamente modificados, incluido el
maíz Bt que produce dos o más toxinas para el control de los taladros, puede reducir la eficacia futura de las
estrategias de manejo de la resistencia.

Key words: insect resistance management, Bt maize, Cry1Ab, corn borers, MON810

© The Author(s) 2023. Published by Oxford University Press on behalf of Entomological Society of America. 275
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/
by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
276 Journal of Economic Entomology, 2023, Vol. 116, No. 2

Current Status of Bt Maize Cultivation in the EU point: Spain, Portugal, France, Germany, Czech Republic, Slovakia,
Poland, and Romania. However, since 2017 only Spain and Portugal
The cultivation of genetically modified (GM) maize producing in-
have planted this crop and of these, only Spain has grown it contin-
secticidal proteins from the bacterium Bacillus thuringiensis (Bt
uously since 1998 (Fig. 1).
maize) has grown exponentially since the first varieties producing
The area under Bt maize cultivation in Spain has been gradually
the Cry1Ab toxin were used in 1996 in the United States and a
increasing to reach about 100,000–125,000 hectares since 2011 (Fig.
year later in Canada to control the European corn borer, Ostrinia
2A) (MAPA 2022a). Currently, it represents 96% of the GM maize
nubilalis (Hübner) (Lepidoptera: Crambidae), and the Southwestern
grown in the EU. The remaining 4% is grown in Portugal, where
corn borer, Diatraea grandiosella Dyar (Lepidoptera: Crambidae),
MON810 varieties have been used commercially since 2005 (DGAV
respectively (Buntin et al. 2004). So far, a total of 210 GM maize
2022). In Spain, it is possible to distinguish three different stages ac-
events conferring insect resistance (181 against Lepidoptera and
cording to the level of adoption: 1) <10% of the total (grain and
126 against Coleoptera) have been developed for commercialization
forage) maize cultivated in the country during 1998–2002; 2) be-
(ISAAA 2019, 2022). Bt maize has been widely cultivated in many
tween 10 and 20% from 2003 to 2010; and 3) between 20 and 30%
countries due to its efficacy against some harmful pests, which makes
from 2011 to 2021 (Fig. 2B) (MAPA 2022a, 2022b). The increase
Bt maize competitive from an agronomic point of view (Dively et
in adoption of Bt maize has been higher in regions where target
al. 2018, Marques et al. 2019) in addition to providing economic,

Downloaded from https://academic.oup.com/jee/article/116/2/275/6968981 by guest on 30 April 2023

pests cause severe and recurrent damage, as in Aragon, Catalonia,
environmental and social benefits (Sanahuja et al. 2011, Brookes
and Navarre in northeastern (NE) Spain (Fig. 2B and C). In Aragon,
2019, Ala-Kokko et al. 2021). In the European Union (EU), the im-
Bt maize adoption gradually increased from less than 30 to 70%,
portation of maize grain from 159 GM maize events producing dif-
between 2003 and 2014; afterwards the adoption rate oscillated
ferent insecticidal toxins is authorized for the placing on the market
between 50 and 60% (Fig. 2B). The increase in use was steeper in
of products containing, consisting of, or produced from this maize
Catalonia than in Aragon, rising from 40 to 70% between 2006 and
(European Commission 2022a, ISAAA 2022). However, only two of
2008, and fluctuating around 70% since 2009 (Fig. 2B). Finally in
these events, both producing Cry1Ab, have been authorized for cul-
Navarre, a slower increase in adoption rate has taken place, from less
tivation to date: Bt176 (Syngenta) from1998 to 2005, and MON810
than 20% in 2004 to around 45% in 2016 and the following years
(Monsanto) since 2003. Maize hybrids derived from event Bt176
(Fig. 2B). In Portugal, Bt maize is mostly grown in the Alentejo region
were withdrawn from the European market in 2006 because this
and its adoption has remained below 25% (Fig. 2C) (DGAV 2022).
event contained an ampicillin resistance gene as selectable marker
The main threat to the long-term success of insect-resistant
(Badosa et al. 2004, Devos et al. 2006). Therefore, since 2006 only
GM crops is the evolution of resistance in target pests (Tabashnik
one GM plant, MON810 maize, has been commercially grown in
et al. 2008, Tabashnik and Carrière 2019, Tabashnik et al. 2022).
the EU. From 2011 to 2021 it was grown on ca.100,000–130,000
Therefore, since the approval of Bt maize cultivation in the EU, Insect
ha annually, accounting for 0.05% of the world area of GM crops in
Resistance Management (IRM) programs have been implemented to
2019 (ISAAA 2019, DGAV 2022, MAPA 2022a).
prevent or delay the evolution of resistance (European Commission
Bt176 and MON810 produce the lepidopteran-specific insec-
2001). These IRM programs include resistance monitoring to assess
ticidal toxin Cry1Ab (Van Frankenhuyzen 2009, Pardo-Lopez
whether susceptibility of target pests to Cry1Ab maize may decline
et al. 2013). The cultivation of these Bt maize hybrids in Europe
over time. Whereas a previous review focused on IRM for Bt maize in
was initially intended for the control of O. nubilalis, but they
Spain, including data produced in our laboratory through 2019 and
were also shown to effectively control another key pest of maize
published without our consent (Álvarez-Alfageme et al. 2022), the
in the Mediterranean area, the Mediterranean corn borer Sesamia
present review updates this topic for the EU by including bioassay
nonagrioides (Lefèbvre) (Lepidoptera: Noctuidae) (González-Núñez
data from our laboratory through 2021 and addresses challenges
et al. 2000, Eizaguirre et al. 2006). Of the 27 countries that currently
and future prospects.
make up the European Union, eight have grown Bt maize at some

Fig. 1. European Union countries that have grown Bt maize (•) (Bt176 and/or MON810 events) since its release in 1998. Sources: Annual Briefs from the
International Service for the Acquisition of Agri-Biotech Applications (ISAAA) (https://www.isaaa.org/resources/publications/briefs/default.asp, accessed: 1 July
2022), MAPA (2022a), and DGAV (2022).
Journal of Economic Entomology, 2023, Vol. 116, No. 2
Spanish Autonomous Communities: 1 (Galicia),
2 (Asturias), 3 (Cantabria), 4 (Basque Country),
5 (Navarre), 6 (La Rioja), 7 (Aragon), 8 (Catalonia),
9 (Castilla-Leon), 10 (Community of Madrid),
11 (Extremadura), 12 (Castilla-La Mancha),
13 (Community of Valencia), 14 (Andalusia),
15 (Region of Murcia).
Portuguese Regions: 16 (Norte), 17 (Centro),
18 (Lisboa), 19 (Alentejo) and 20 (Algarve).
Fig. 2. Bt maize in the Iberian Peninsula (Spain and Portugal). (A) Area of Bt
maize (Bt176 and MON810) in Spain from its introduction in 1998 to 2021. (B)
Bt maize adoption rate in Spain and the three Autonomous Communities in
NE Spain: Aragon, Catalonia and Navarre. (C) Bt maize adoption rate per area
in the Iberian Peninsula in 2021. Sources: MAPA 2022a, b, DGAV 2022.
Target Pests of Bt Maize in the EU
Insect pests are a major cause of economic losses in global maize
production (Bradshaw et al. 2016, Manosathiyadevan et al. 2017),
with several lepidopteran and coleopteran key pests limiting the
yield of this crop (Edde 2022). In Europe, two of the most de-
structive pests of maize are the corn borers S. nonagrioides and
O. nubilalis, which are the target pests of Cry1Ab maize. Larvae
277
of both moths penetrate inside the maize stalks, where they exca-
vate galleries and feed on the plant stem. This causes the stalks to
break and the ears to fall, resulting in reduced yields. Stalk damage
from both species is very similar (Eizaguirre and Fantinou 2012),
but S. nonagrioides larvae are more voracious (Butrón et al. 1999,
Velasco et al. 1999). Yield reductions also result from O. nubilalis
larval feeding on kernels later in the season (Andreadis et al. 2008).
In addition, damage from both corn borers promotes mycotoxigenic
fungal growth, such as Fusarium spp. Link, which cause two of the
most common forms of maize ear rot in Europe, the red ear rot and
the pink ear rot (Logrieco et al. 2002). Mycotoxin contamination of
maize grain is an important problem associated with the presence of
corn borers and fungal growth, reducing crop quality and increasing
the risks associated with food and feed production (Pitt et al. 2013,
Arias-Martín et al. 2021). It has been estimated that between 2.25
Downloaded from https://academic.oup.com/jee/article/116/2/275/6968981 by guest on 30 April 2023
and 4 million hectares of maize are directly affected annually by corn
borers in Europe (Brookes 2019), causing annual average yield losses
that range from 5 to 30% (Meissle et al. 2010). Knowledge about
the biology and ecology of these target pests is essential to optimize
the use of Bt maize and to design efficient IRM plans to avoid rapid
evolution of resistance. However, there are still gaps in knowledge
to be filled, especially in the case of S. nonagrioides (Castañera et
al. 2016).
S. nonagrioides is distributed across the Mediterranean basin, in-
cluding Southern Europe, the Middle East, and countries in North
and Western Africa (Eizaguirre and Fantinou 2012), where it is one
of the most damaging pests of maize (Anglade 1972, Melamed-
Madjar and Tam 1980, Castañera 1986). It is a polyphagous spe-
cies with host plants in the Poaceae, Cyperaceae, and Thyphaceae
families (Kfir et al. 2002, Le Rü et al. 2006). Premating movement
is rare among females and mating occurs near adult emergence sites,
which might have important implications for Bt resistance manage-
ment (Eizaguirre et al. 2004). After mating, females deposit eggs
clutches between the leaf sheath and the stalk (López et al. 2003).
First instar larvae begin to excavate galleries towards the interior of
the plant, which makes larval control with conventional insecticides
difficult (Velasco et al. 1999). Larvae usually molt 5 times during
development (López et al. 2001), reaching 3.5–4 cm in their last in-
star (Anglade 1972). Movement among plants has been reported for
last instar larvae (Eizaguirre et al. 2004). The number of generations
per year depends on the climatic zone, latitude, and temperature.
Two to three generations have been recorded in Spain and France
(Anglade 1972, Galichet 1982, Eizaguirre et al. 2002) and up to four
in Morocco and Portugal (Hilal 1977, Figueiredo and Araújo 1996).
O. nubilalis currently occurs in Europe, North Africa, West Asia,
and North America. It is a highly polyphagous species feeding on
more than 200 wild plants. It colonized maize after this crop was
introduced to Europe about 500 years ago (Gould 1998, Marçon
et al. 1999a, Bethenod et al. 2005). Adults can disperse tens of km
in just a few nights (Showers et al. 2001). After emergence some
adults move locally but others enter long-range dispersal (Bourguet
et al. 2000a, b, Bailey et al. 2007, Gaspers 2009). Predispersal
mating likely occurs for both males and females (Dalecky et al.
2006, Dorhout et al. 2008, Hu and Andow 2011). Females lay
clutches of eggs on the underside of leaves and early larval instars
feed on developing leaves and whorl tissues (Gaspers 2009). Larvae
do not penetrate the stalk until the beginning of the third instar,
and once inside feed on the meristematic tissue of the stem, digging
galleries in the stalk (Mason et al. 1996, Mencarelli et al. 2013). In
Europe, univoltine populations have been reported in Germany and
the Netherlands, whereas in warmer regions of southern Europe,
multivoltine populations are present (Gaspers 2009, Keszthelyi
278
2010). Two pheromone races (E- and Z-) have been identified: the
E-race is found in Switzerland, Italy and Eastern North America; and
the Z-race predominates over most of Europe and North America
(Marçon et al. 1999a). Both races are present in Spain, although the
Z-race is predominant (Gaspers 2009). Interestingly, a study in the
USA has shown that susceptibility of this species to Cry1Ab does not
differ between pheromone races, voltinism ecotypes or geographical
areas (Marçon et al. 1999b).
Other noctuid moths are occasional pests of maize in Europe
but are not considered target pests of Cry1Ab maize. Two of the
most important are Mythimna unipuncta (Haworth), which can
cause severe defoliation and Helicoverpa armigera (Hübner), which
can damage silk and cob tips (Pérez-Hedo et al. 2012). Both species
have low susceptibility to Cry1Ab (Eizaguirre et al. 2010, González-
Cabrera et al. 2013, Pérez-Hedo et al. 2013).
Insect Resistance Management for Bt Maize in
the EU
Since commercialization of Bt maize in the EU, IRM plans have
been implemented to prevent or delay the evolution of resistance to
Cry1Ab (Head and Greenplate 2012). IRM plans are based on the
high-dose/refuge (HDR) strategy, which is commonly used world-
wide and mandatory in the EU (EFSA 2018). This strategy is based
on cultivation of Bt crops that produce a high concentration of in-
secticidal proteins effective against the targeted pests and the deploy-
ment of refuges (non-Bt crop host plants) near Bt crops (Siegfried
and Hellmich 2012, Tabashnik et al. 2013). In many countries this
strategy is currently based on use of pyramided Bt crops that pro-
duce more than one toxin effective against the same target pest
(Bates et al. 2005, Carriére et al. 2015, 2016, Van den Berg et al.
2022). However, this strategy cannot be implemented in the EU since
only MON810 varieties that produce Cry1Ab are approved for cul-
tivation (European Commission 2022a). An important part of IRM
plans is the implementation of a monitoring program that enables
early detection of resistance evolution, which may allow the applica-
tion of corrective measures in a timely manner if resistant populations
are detected (Andow and Ives 2002, Head and Greenplate 2012). All
European Bt maize is concentrated in the Iberian Peninsula, mostly
in Spain. Thus, the EU resistance monitoring plans for pests targeted
by Bt maize have focused primarily on this country (Farinós et al.
2018, Thieme et al. 2018, Bertho et al. 2020), particularly northeast
Spain since 2016 (EFSA 2017).
Success of the HDR strategy used to delay evolution of resist-
ance to single-toxin Bt crops can be improved by several conditions,
including: 1) recessive inheritance of resistance; 2) most matings
between the rare individuals that survive on the Bt crop should be
with susceptible individuals from refuges; and 3) resistance allele
frequencies in field populations should be low (Roush 1994, Bates et
al. 2005, Tabashnik et al. 2013). Violations of any of these conditions
can increase the likelihood of resistance evolution (Gassmann et al.
2011, Tabashnik et al. 2013, Tabashnik and Carrière 2019, Huang
2021).
Bt plants should produce a sufficient concentration of toxin
to kill heterozygote individuals carrying a single resistance allele
(Carrière et al. 2010). As a consequence, only homozygous resistant
individuals would survive on Bt plants, and mate with the more
abundant susceptible individuals produced in refuges (Andow 2008).
Their offspring would be heterozygote, and thus susceptible to Bt
plants (Bourguet et al. 2000c). Maize varieties carrying the event
MON810, which are the only GM varieties grown in Europe, pro-
duce high levels of Cry1Ab throughout their development (Székács
Journal of Economic Entomology, 2023, Vol. 116, No. 2
et al. 2010a,b), which is enough to kill 100% of susceptible larvae
of both O. nubilalis and S. nonagrioides (Farinós et al. 2018). Thus,
it is assumed that they fulfill the high dose requirement, though no
Cry1Ab resistant populations of O. nubilalis nor S. nonagrioides
have been detected to determine if resistance in these species is ef-
fectively recessive. By contrast, maize hybrids derived from event
Bt176 cultivated between 1998 and 2005 showed a decrease in
toxin concentration during plant development (Fearing et al. 1997).
This resulted in larvae being exposed to lower concentrations of the
toxin, which could have favored the evolution of resistance (Archer
et al. 2000, Magg et al. 2001).
Both in Portugal and Spain, mandated refuges must be at least
20% of the area planted with Bt maize. Conventional maize that
has a similar phenology as Bt maize should be used in refuges, since
maize is the main host of both O. nubilalis and S. nonagrioides
Downloaded from https://academic.oup.com/jee/article/116/2/275/6968981 by guest on 30 April 2023
(Bourguet et al. 2000b, Camargo et al. 2020). Refuges can be sown
in the same field or in adjacent fields at a maximum distance of
750 m, because of limited dispersal of S. nonagrioides short flight
patterns. In Spain, refuges are not required for growers who plant
less than 5 ha of Bt maize. In Portugal, planting refuges is always
mandatory. Refuge compliance in Spain has been above 80% since
2008 and above 90% since 2015, while in Portugal it has been
100% according to questionnaires sent to growers (Farinós et al.
2018, European Commission 2022b).
The frequency of alleles conferring resistance to MON810
maize was estimated in EU field populations of O. nubilalis and S.
nonagrioides between 2003 and 2005. Resistant allele frequencies
were 0.0001 (CI 95% 0.0–3.0 × 10−4) for O. nubilalis (Engels et
al. 2010) and 0.0015 (CI 95% 0.0–4.6 × 10−3) for S. nonagrioides
(Andreadis et al. 2007).
Resistance Monitoring Plans for Bt Maize in
the EU
Monitoring plans in the EU aim to detect declines in suscepti-
bility of the targeted pests to Cry1Ab in areas where Bt maize is
grown. These plans are mandatory since 2001, when the European
Parliament Directive 2001/18/CE of March 12th was published
(European Commission 2001). However, the Spanish Central
Administration started their own monitoring plan three years be-
fore this directive and this was implemented until 2011 (MITECO
2010). Complementary to this effort, seed companies have had
their own monitoring plan since 2004 (Farinós et al. 2018, Thieme
et al. 2018). The EU monitoring plans have focused mainly on the
Iberian Peninsula (Spain and Portugal) where adoption of Bt maize
has been higher. Monitoring was also carried out in some years be-
tween 2000 and 2010 in southwestern France (Midi-Pyrénées and
Poitout-Charentes) for S. nonagrioides and in the Czech Republic,
France, Germany, Italy, Hungary, Slovakia, Poland, and Romania for
O. nubilalis (Farinós et al. 2018, Thieme et al. 2018).
Results of the monitoring performed by the Spanish
Administration between 1999 and 2011 and by seed companies
between 2004 and 2015 have been published (Farinós et al. 2004,
2011, 2018; Castañera et al. 2016, Thieme et al. 2018, Bertho et al.
2020). In Spain and Portugal they focused on the northeast (Ebro
Valley, Spain), center (Madrid and Castilla-La Mancha, Spain), and
southwest (Alentejo, Portugal; and Extremadura and Andalusia,
Spain) (Fig. 2C). Sampling in these areas was carried out every other
year. Susceptibility assessment relied on concentration-response
diet-overlay bioassays with purified Cry1Ab to estimate lethal
concentrations (LC50) or molt inhibition concentrations (MIC50)
affecting 50% of the population. The results indicated no shifts
Journal of Economic Entomology, 2023, Vol. 116, No. 2
over time in the susceptibility of S. nonagrioides or O. nubilalis
to Cry1Ab. In France, where MON810 hybrids were cultivated in
2005–2007, S. nonagrioides variation in both MIC50 and LC50 was
in the range observed for Iberian populations (Farinós et al. 2018).
Likewise, only small variation in susceptibility (MIC50) to Cry1Ab
was observed for European O. nubilalis populations from other ge-
ographic regions (Thieme et al. 2018).
Update on the Status of the Monitoring Program
In 2016, the monitoring plan changed in accordance with the data
gathered for more than a decade and the recommendations of
the European Food Safety Authority (EFSA 2015, 2016a). Since
then, a stepwise approach has been followed (Fig. 3, approach for
S. nonagrioides). The aims of the changes were to strengthen the
efforts in areas where resistance is more likely to evolve and to
reach a minimum detection threshold for resistance allele frequency
of 3%. Thus, insect collection has been focused on hotspots, i.e.,
areas where adoption rate of Bt maize surpasses 60% and targeted
pests have two or more generations yearly (EFSA 2015, 2016a). In
the EU, this situation only occurs in NE Spain, where monitoring is
now carried out annually. Diagnostic concentration (DC) bioassays
with Cry1Ab, intended to cause ≥99% molting inhibition for first
instar larvae (MIC99), and plant bioassays with Cry1Ab maize are
used to measure susceptibility of both species. The bioassays for S.
nonagrioides are performed by our group whereas the laboratory
BTL GmbH Sagerheide (Germany) carries out bioassays for O.
nubilalis. An annual report is sent by the seed companies to EFSA
for evaluation, revision, and modification when necessary. These
reports have been available since 2009 at the EC website (European
Commission 2022b).
Between 2016 and 2021 no significant shifts were observed in
the susceptibility of NE Spain field populations of S. nonagrioides
to Cry1Ab in DC bioassays (Fig. 4A) or plant bioassays (Fig. 4B).
During the DC bioassays the susceptibility of the field-collected
larvae is checked against both a theoretical molt inhibition value
of 99% and the molt inhibition displayed by the reference strain.
Statistically significant differences were detected in 2017 and 2019
between the observed and the expected MIC99 values obtained in
DC bioassays (Fig. 4A). In 2017 the observed MIC99 value of the
field populations was significantly lower than that of the laboratory
strain (Fig. 4A). In plant bioassays, the mortality of first instar larvae
from field collected populations was 99.9% in 2017, the first year
this assay was performed, and 100% in subsequent years (Fig. 4B).
Throughout these years, no clear trends are observed, suggesting
that the differences noticed between some of the years could be at-
tributable to natural variation in the field populations. Likewise,
no decrease in the susceptibility to Cry1Ab has been detected in O.
nubilalis (European Commission 2022b). These results confirm that
MON810 maize is still effective against both species and there are
no signs of field evolved resistance.
Assessment of the Implementation of the Current
Monitoring Plan
A critical reassessment of the monitoring plan carried out in
2004–2015 has already been accomplished (Farinós et al. 2018).
Recommendations implemented since 2016 (EFSA 2015) include fo-
cusing on target pest populations from hotspots and reducing the
detection limit for resistance allele frequency from 5 to near 3%.
However, the experience accumulated through the implementation
of this new monitoring plan has revealed some practical and tech-
nical limitations, which are addressed below.
279
Sources of Agronomic Data
The use of reliable and thorough agronomic databases is essential
to identify areas with the greatest selection pressure to be incorpo-
rated in resistance monitoring plans. However, in Spain, there are
notable differences in the estimated areas of Bt maize cultivated an-
nually, depending on the administration that generates these data.
The acreage data provided by the Spanish Ministry of Agriculture
is an indirect measure obtained from the number of seeds sold, pro-
vided by the companies, and the maize planting density. However,
taking as an example Catalonia, one of the regions with the highest
level of adoption of Bt maize, data on cultivated areas are also pro-
vided by the local Government based on questionnaires answered
by farmers (Fig. 5). This may give rise to inconsistencies between
estimates provided by the Spanish Ministry of Agriculture and local
Governments that complicate decision making. Other data needed
Downloaded from https://academic.oup.com/jee/article/116/2/275/6968981 by guest on 30 April 2023
for the monitoring plan, such as the area of Bt maize dedicated to
grain or feed maize, or first or second harvest maize, which varies
depending on regions, are sometimes unavailable or difficult to
access.
Insect Collection
Last-instar larvae of S. nonagrioides and O. nubilalis are collected
for bioassays following standard operative procedures (EuropaBio
2017), from three sampling zones in NE Spain each year, each with
a maximum 10 km diameter. The goal is to collect a minimum of
1,000 larvae of each species since, to achieve a 3% detection limit
for resistant alleles, this is the number required for larval screens
assuming recessive alleles (1/N1/2, where N is the number of larvae)
(Andow and Ives 2002). The protocol establishes the collection of at
least 350 larvae from three fields, with a minimum of 50 larvae per
field (successful field) within each sampling zone. This objective has
been met for S. nonagrioides for each campaign (Table 1). However,
since 2016 it has proven difficult to meet this requirement for O.
nubilalis, and it has not been possible in the last two campaigns
(Table 1). Additionally, the targeted number of successful fields
(three in each zone) has not been met in any of the campaigns for O.
nubilalis even though the number of prospected fields has increased
annually (Table 1).
The difficulties encountered in collecting enough O. nubilalis
larvae could be related to regional pest suppression due to continued
Bt maize cultivation in NE Spain since 1998. Regional pest suppres-
sion through high adoption of Bt maize has been reported for O.
nubilalis in the United States (Hutchison et al. 2010, Bohnenblust et
al. 2014) and for several other pests (e.g., Carrière et al. 2003, Wu et
al. 2008). Such regional declines may provide important agronomic
benefits, for farmers growing Bt maize, and other types on non-Bt
crops (Hutchison et al. 2010, Dively et al. 2018). However, the lack
of available historical records of damage caused by the corn borer or
population density of this pest in Spain does not allow us to verify
that regional declines have occurred. Additional studies are needed
to determine if declines in the densities of this pest have occurred
in areas where they have historically caused important damage to
maize.
Insect Rearing
Diapausing larvae of both species are usually collected, and kept in
this condition until they emerge from diapause. They are then reared
until pupation and subsequent adult emergence for mating. Some
of the individuals collected are lost during rearing, mainly in the
larval stage, due to pathogens or parasitoids (mostly Tachinidae in
the case of S. nonagrioides), although there is also mortality due to
malformations in pupae and adults (Supp Fig. 1 [online only]). This
280
Fig. 3. Flowchart of the monitoring strategy for S. nonagrioides resistance to MON810 maize in the EU.
mortality, together with the infertility of some adults, means that
not all individuals collected in the field are represented in bioassays
(Table 1).
S. nonagrioides preimaginal mortality for larvae collected in
the period 2016–2021 was 42 ± 5% with relatively high variability
observed between years (from 30% in 2016 to 61% in 2018). Such
larval mortality has hindered the resistance allele detection limit,
Journal of Economic Entomology, 2023, Vol. 116, No. 2
Downloaded from https://academic.oup.com/jee/article/116/2/275/6968981 by guest on 30 April 2023
only reaching the target 3% in the 2021 campaign, even though the
number of larvae collected had increased over the years (Table 1).
Optimization of several procedures to reduce this preimaginal mor-
tality have been tried and implemented, such as conducting larvae
disinfection, reduction in the number of larvae per rearing box, re-
duction in the number of times per week that the rearing boxes are
checked to avoid excessive handling and increasing the change of
Journal of Economic Entomology, 2023, Vol. 116, No. 2
Fig. 4. Molting inhibition of S. nonagrioides larvae coming from NE Spain
field populations (black columns) or from a susceptible laboratory strain (grey
columns) from 2016 to 2021. (A) Diagnostic concentration (DC) bioassays
using Cry1Ab, intended to cause ≥99% molting inhibition to first-instar larvae
(MIC99, dash line), (B) Plant bioassays with Bt maize. Source: https://food.
ec.europa.eu/plants/genetically-modified-organisms/post-authorisation/
monitoring-plans-and-reports_en#pmem-reports-for-gm-food-and-feed
vermiculite during the diapause period, among others. However, no
significant improvements in preimaginal survival rates have been
observed.
Similar problems have been experienced by the BTL GmbH
Sagerheide laboratory that assesses the susceptibility of O. nubilalis
field populations collected in Spain, which has not been able to re-
duce the allelic detection limit for this species below 4% (European
Commission 2022b).
Laboratory Strains
It has proven difficult to maintain reference strains of some lepi-
dopteran species for many years in the laboratory, mainly because
281
50.000
Bt maize area in Catalonia (ha)
40.000
30.000
20.000
10.000
2017
1999
2001
2003
2005
2007
2009
2011
2013
2015
2019
2021
Government of Spain (mapa.es)
Downloaded from https://academic.oup.com/jee/article/116/2/275/6968981 by guest on 30 April 2023
Government of Catalonia (gencat.cat)
Fig. 5. Differences in the Bt maize area reported by different Spanish
authorities (MAPA 2022a, GENCAT 2022).
they suffer excessive inbreeding (Roush 1986), but also due to
pathogens accidentally introduced in the laboratory by field col-
lected populations. To tackle this issue, the reference strains of
S. nonagrioides and O. nubilalis are refreshed periodically with
specimens collected in non-Bt fields from Galicia (northwestern
Spain), where Bt maize has never been commercially grown. Even
so, the performance of laboratory strains can still decline over
time, as the strains often demonstrate decreasing larval weight and
adult fertility. In those cases, new reference strains are generated
with individuals collected in Galicia, taking some precautions:
1) comparison of the LC50 values of both laboratory (Fig. 6) and
field populations, estimated by concentration-response diet-overlay
bioassays with purified Cry1Ab (Farinós et al. 2018), to guarantee
that there are no significant differences between them; 2) check for
presence of pathogens (namely Nosema sp.) by microscopy and by
molecular methods (PCR); and 3) ensure that the new population is
adapted to feeding on an artificial diet under laboratory conditions.
Why has Resistance not Evolved? Sustained
Efficacy of Bt Maize against S. nonagrioides
The evolution of resistance of field populations to Bt maize producing
Cry toxins has occurred in at least six lepidopteran species to date.
Four of them are noctuids (Lepidoptera: Noctuidae): Busseola fusca
(Fuller) in South Africa, the fall armyworm Spodoptera frugiperda
(JE Smith) in the United States, Brazil, and Argentina, Helicoverpa
zea (Boddie), in the United States and Striacosta albicosta Smith
in the United States and Canada; and two crambids (Lepidoptera:
Crambidae): Diatraea saccharalis (Fabricius) in Argentina and O.
nubilalis in Canada (Smith et al. 2019, Tabashnik and Carrière
2019, Tabashnik et al. 2022). Such field-evolved resistance has been
attributed to a variety of causes, including exposure of the larvae to
low concentrations of the toxin (B. fusca, H. zea, S. frugiperda, and
S. albicosta) which may yield nonrecessive inheritance of resistance
(Tabashnik et al. 2013), high adoption rate and repeated cultivation
(B. fusca, H. zea, S. frugiperda, S. albicosta, D. saccharalis, and O.
nubilalis), low refuge compliance (B. fusca, H. zea, S. frugiperda,
D. saccharalis, and O. nubilalis), and lack of long-range migration
from outside agricultural systems due to geographic or agronomic
characteristics, which limits genetic variability (B. fusca, H. zea, and
S. frugiperda) (Van Rensburg 2007, Dively et al. 2016, Omoto et
al. 2016, Ostrem et al. 2016, Chandrasena et al. 2017, Smith et al.
2017, Campagne et al. 2013, Grimi et al. 2018, Smith et al. 2019).
282 Journal of Economic Entomology, 2023, Vol. 116, No. 2

Table 1. Collection of S. nonagrioides and O. nubilalis larvae and resistance allele detection limit for S. nonagrioides, corresponding to the
European MON810 maize resistance monitoring plan carried out from 2016 to 2021

2016 2017 2018 2019 2020 2021

S. nonagrioides Larvae collected 1,364 1,452 1,490 1,644 1,590 1,699

Prospected fields 29 19 18 28 27 24
Successful fieldsa 11 9 9 9 11 12
Larvae per field 47.0 76.4 82.8 58.7 58.9 70.8
Adults represented in bioassaysb 911 (67%) 749 (52%) 554 (37%) 868 (53%) 787 (50%) 1076 (63%)
Resistance allele detection limit 3.3% 3.7% 4.3% 3.4% 3.6% 3.0%
O. nubilalis Larvae collected 1,111 1,111 1,144 1,110 651 811
Prospected fields 17 19 23 28 27 29
Successful fieldsa 6 5 7 8 4 5
Larvae per field 65.4 58.5 49.7 39.6 24.1 28.0

a
Successful fields are those where at least 50 larvae were collected.

Downloaded from https://academic.oup.com/jee/article/116/2/275/6968981 by guest on 30 April 2023

b
Number of adults, reared from larvae collected in the field, whose progeny were used in the bioassays. The percentage with respect to the total
number of larvae is indicated among brackets.

A) S. nonagrioides B) O. nubilalis
100 100
90 90
80 80
70 70
Mortality (%)

Mortality (%)
60 60
2016
50 50 2017
40 40 2018
30 30 2019
20 20 2020
10 10 2021
0 0
0,1 1,0 10,0 100,0 1000,0 10000,0 0,1 1,0 10,0 100,0
Concentration (ng/cm2) Concentration (ng/cm2)

The results of molting inhibition of laboratory populations at different concentrations of Cry1Ab

(dose-response bioassays) were adjusted by probit weighted regression lines using POLOPlus 1.0
(LeOra Software, 2002-2022).
Fig. 6. Regression lines of mortality for laboratory strains of S. nonagrioides (A) and O. nubilalis (B) between 2016 and 2021. Source: https://food.ec.europa.eu/
plants/genetically-modified-organisms/post-authorisation/monitoring-plans-and-reports_en#pmem-reports-for-gm-food-and-feed

The corn borer S. nonagrioides presents certain biological
characteristics that could accelerate the evolution of resistance to
Bt maize: 1) despite being a polyphagous species, it is considered
oligophagous or largely monophagous in maize areas (Castañera
et al. 2016, Camargo et al. 2020); 2) multivoltine populations
may have increased exposure to Bt maize (Eizaguirre et al. 2002);
3) adults, especially those of the third generation, have low dis-
persal between refuges and Bt maize (Eizaguirre et al. 2006) and
there is a low genetic exchange between distant populations (De
La Poza et al. 2008); and 4) females mate before they disperse to
oviposit (Lopez et al. 1999), so adults from refuges would rarely
mate with the moths that eventually emerge from Bt maize. There
are also a number of agronomic characteristics associated with
the cultivation of Bt maize in NE Spain that could also accel-
erate evolution of resistance in this area, such as high adoption
rate of Bt maize in the region and continuous cultivation of Bt
maize. Altogether, the biological and agronomic traits define the
region as a resistance hotspot for S. nonagrioides (EFSA 2018).
However, no resistant populations have been detected (European
Commission 2022b).
To determine what has delayed the evolution of resistance of S.
nonagrioides to Cry1Ab maize, a resistance evolution model was de-
veloped (Castañera et al. 2016). The model includes three different
types of parameters: 1) population parameters (proportion of adults
leaving fields; relative preference for Bt and non-Bt fields; adult fe-
cundity and survival; diapause rate and overwintering survival); 2)
genetic parameters (initial R allele frequency; RR, RS and SS survival
on each type of maize and in each generation; assortative mating in
each generation); and 3) environmental and agronomical parameters
for the period 1998 and 2013 (proportion of each type of maize,
i.e., Bt maize (event Bt176 or MON810) versus non-Bt maize; pro-
portion of maize rotated to another crop; refuge compliance). This
model predicted that, if current cultivation conditions continue, the
resistance allele frequency would be >0.5 by 2050 in the Ebro Valley
(Castañera et al. 2016). The modeling results indicate withdrawal of
Bt176 from the market was a critical factor in delaying the evolu-
tion of resistance because of the low concentration of Cry1Ab late
in the season in this event. They also emphasized the importance of
compliance with the planting of refuges in areas with high adoption
rate of Bt maize such as the Ebro Valley, and for refuges to be located
Journal of Economic Entomology, 2023, Vol. 116, No. 2
close to Bt maize fields to enhance mating between resistant and sus-
ceptible adults. Currently, the guide of good practices for planting Bt
maize in Spain has established this distance at <750 m (MAPA 2015,
European Commission 2022b).
The F2 screen method (Andow and Alstad 1998) was used twice
to determine the frequency of resistance alleles in field populations
of S. nonagrioides from southern Europe: 1) in 2004–2005 in
NE Spain (Ebro Valley), after 7–8 years of Bt maize cultivation
and with an adoption rate around 30%, and in Greece, where Bt
maize has never been grown; and 2) in 2016, again in the Ebro
Valley, after 18 years of cultivation and with a mean adoption
rate of 64%. The frequency of resistance alleles in the southern
European populations was estimated as 0.0015 (0.0032 for Greece
and 0.0029 for Ebro Valley) in 2004–2005 (Andreadis et al. 2007).
These results reflect the absence of isolines giving positive results
(survivors at F2) although only 85 isolines were tested from the
Ebro Valley. In 2016, 1 of 137 isolines was scored as positive for
resistance, yielding an estimated resistance allele frequency of
0.0036 (Camargo et al. 2018). This new value was used to up-
date the existing model (Andreadis et al. 2007). The results indi-
cate that resistance would occur in 31 years from 2016, 2.8 years
earlier than expected with the allelic frequency of 0.0029 estimated
in 2004–2005 (Castañera et al. 2016, Camargo et al. 2018). The
estimated resistance allele frequency was not significantly higher
in 2016 than 2004–2005. The monitoring and modeling results
imply the HDR strategy could be effective in delaying the evolution
of resistance in the NE Spain if strict compliance with the refuge
requirements continues.
Gaps that need to be filled to confirm key assumptions and fur-
ther refine model results are mainly related with pest’s biology (as-
sortative mating and use of other hosts at certain times of the year
or in certain zones, etc.) (Castañera et al 2016). Moreover, global
warming may change some factors such as the length of the crop
season.
Challenges and Future Prospects
The year 2022 is the 25th year of continuous planting of Bt maize
in Spain, and the 19th year since planting of varieties derived from
the event MON810 began. To date, the monitoring data show that
the target pests S. nonagrioides and O. nubilalis have not evolved
resistance to Cry1Ab in the field. Also, seed marketing companies
have not received reports of confirmed control failures (European
Commission 2022b). The efficacy of MON810 in controlling these
pests and compliance with the mandatory refuge planting in re-
sistance hotspots, especially during the last fifteen years (both the
companies marketing GM seeds and the competent authorities have
made great informational efforts) may have contributed to this suc-
cess. However, the EU is facing several challenges that may jeop-
ardize the planting of GM crops in the future.
The main challenge for the cultivation of Bt maize in the EU is
social rejection in many countries. As a consequence, new events
cannot be readily approved in the EU, despite their potential ag-
ronomic, economic, social, and environmental benefits (Gomez-
Barbero et al. 2008, Brookes 2019, Areal and Riesgo 2022).
New authorizations of GMOs require approval by a qualified
majority of Member States in the Council of the EU, yet many
countries oppose cultivation of GMOs (Tagliabue 2017, Hundleby
and Harwood 2019). This situation is not likely to change soon
(Eriksson et al. 2019). Also, in some situations, farmers face ec-
onomic losses or increased labor to comply with coexistence
regulations, i.e., legal requirements imposed by the European
283
Commission to ensure GM crops are not mixed with non-GM
crops at the farm level (European Commission 2010 , Park et al.
2011, Kathage et al. 2016).
The limitations in the EU for the approval of new GM crops are
a serious constraint for improving resistance management strategies,
because pyramided Bt maize hybrids producing several Bt toxins are
not allowed. The simultaneous use of multiple toxins that do not
exhibit cross-resistance is one of the main tactics recommended for
delaying insect resistance to Bt crops (Roush 1998, Carrière et al.
2015, 2016). This approach is based on the fact that if resistance
to each toxin in a pyramid is rare, individuals with resistance to all
toxins will be extremely rare (Tabashnik 1994). A large number of
pyramided Bt maize hybrids are available (ISAAA 2022), which can
improve control of maize pests and resistance management (Carrière
et al. 2015, 2016; Marques et al. 2019). Pyramids have strong po-
Downloaded from https://academic.oup.com/jee/article/116/2/275/6968981 by guest on 30 April 2023
tential to enhance resistance management for O. nubilalis (Tan et
al. 2013) and would likely be useful to improve management of
resistance for S. nonagrioides, since laboratory studies show high
susceptibility of this species to Cry1Ab and Cry1Fa and potentially
low cross-resistance between these toxins (González-Cabrera et al.
2006).
The current situation in the EU regarding the difficulty of
approving new GM crops is also constraining control of inva-
sive and emerging key pests of maize, such as the Western corn
rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera:
Chrysomelidae). The advance in Europe of D. v. virgifera is a
matter of growing concern. Originally from Central America,
this species now occurs in more than 15 European countries, in-
cluding France, Italy, Greece, Poland, Czech Republic, Hungary,
Romania, and Croatia (Mrganić et al. 2018, Bažok et al. 2021).
Diabrotica v. virgifera has also been recently reported in Spain in
Aragon and Catalonia in the NE of the country (Mateu et al. 2021,
CSCV 2021). In the United States where D. v. virgifera is a major
pest of maize (Gassmann et al. 2011), one of the main tools for its
control has been pyramided Bt maize that will be combined with
use of RNA interference (RNAi) starting in 2022 (Carrière et al.
2020, Meinke et al. 2021). In Europe, the level of damage caused
by this pest varies depending on the area and its control is based
on crop rotation, as the cultivation of Bt maize varieties effective
against this coleopteran pest is not authorized (Bažok et al. 2021).
However, this beetle has so far overcome different management
practices, including crop rotation, conventional insecticides, and Bt
maize (Wright et al. 2000, Levine et al. 2002, Carrière et al. 2020,
Gassmann 2021). Since 1995, monitoring by European countries
has measured population fluctuations of D. v. virgifera (Bažok et
al. 2021).
There is also concern that S. frugiperda may become established
in continental Europe. Because this pest has low susceptibility to
Cry1Ab, MON810 would not provide efficient control (Storer et
al. 2010, Omoto et al. 2016). This key pest of maize has already
evolved resistance to different Bt toxins in several populations in
the Americas (Tabashnik and Carrière 2019, Tabashnik et al. 2022).
This pest is a highly migratory species that has recently colonized
most countries south of the Sahara and Southeast Asia, including
India, and China, from where it has reached Australia (Goergen et
al. 2016, EPPO 2022). It has also been recently introduced into the
Canary Islands (Spain) (Moreno and Gastón 2020) from the African
continent (EPPO 2021). Some forecasting models predict that
southern and central Europe could be exposed to the risk of tran-
sient populations, especially with global warming (Tepa-Yotto et al.
2021, Gilioli et al. 2022). Also, there has been an increasing number
of interceptions (33 in 2021 alone) of this species in plants and plant
284
products imported into the EU from both the Americas and Africa
(EUROPHYT 2022).
A recent issue of concern in the EU is the expansion of teo-
sinte, the closest wild relative of maize. This invasive species was
reported as a new agricultural weed in France in 1990 and in Spain
in 2014. Its distribution has increased despite efforts to contain its
advance (Pardo et al. 2016, Le Corre et al. 2020). Different agro-
nomic and environmental consequences of hybridization between
maize and teosinte have been identified, including the transfer of
the cry1Ab transgene from Bt maize to teosinte, which could expose
corn borers to low concentrations of the toxin and accelerate the
evolution of resistance. The presence of the cry1Ab transgene has
been demonstrated in different genetic backgrounds derived from
Bt maize and teosinte crosses, giving rise to plants producing sim-
ilar Cry1Ab concentrations as in the Bt parental hybrid (Erasmus
et al. 2019; Lohn et al. 2021). However, mortality of O. nubilalis
was sometimes less than 99% (Lohn et al. 2021). Although some
uncertainties remain (EFSA 2022), EFSA (2016b) concluded there is
no need to revise current risk management recommendations for Bt
maize. However, further studies are warranted to assess the poten-
tial impact of teosinte on evolution of resistance to Bt maize in corn
borers. These studies could evaluate factors such as the frequency of
hybridization under field conditions with Bt maize varieties, the suit-
ability of teosinte and hybrid progeny as host plants for corn borers
and other nontarget lepidopterans, and the concentration of Cry1Ab
in teosinte and hybrid progeny after several generations of crossing.
Acknowledgments
This review includes the revision of data obtained by our group in
the frame of various R&D contracts of technological support funded
by private companies: Monsanto Europe SA (2016–2019) and Bayer
CropScience Schweiz AG (since 2020). We gratefully acknowledge
the support of field and laboratory technicians, who have contributed
substantially to the search of maize fields, collection of larvae, and
maintenance of insect colonies. We thank Monsanto Company/ Bayer
R&D Services LLC (St. Louis, MO, US) for providing the Cry1Ab
protein, and Bayer CropScience and Pioneer Hi-Bred for providing
Bt maize seeds. Finally, we would like to acknowledge the insightful
comments and suggestions by two reviewers who contributed to
the significant improvement of the manuscript. This manuscript is
published with the concurrence of the Bayer Group, legal owner of
the bioassay data presented in Figures 4 and 6 and Table 1, which
were generated in our laboratory. Data from 2016 to 2020 are in-
cluded in the annual insect resistance monitoring reports associated
with MON810 maize cultivation in the EU, available at the webpage
of the European Commission (https://food.ec.europa.eu/plants/
genetically-modified-organisms/post-authorisation/monitoring-
plans-and-reports_en#pmem-reports-for-gmo-cultivation). Data
for 2021 were not published previously. Any opinions, conclusions
or recommendations expressed in this publication are those of the
author(s) and do not necessarily reflect the view of funding companies.
Supplementary Data
Supplementary data are available at Journal of Economic
Entomology online.
References Cited
Ala-Kokko, K., L. L. Nalley, A. M. Shew, J. B. Tack, P. Chaminuka, M. D.
Matlock, and M. D’Haese. 2021. Economic and ecosystem impacts of GM
maize in South Africa. Glob. Food Sec. 29: 100544.
Journal of Economic Entomology, 2023, Vol. 116, No. 2
Álvarez-Alfageme, F., Y. Devos, A. M. Camargo, S. Arpaia, and A. Messéan.
2022. Managing resistance evolution to transgenic Bt maize in corn borers
in Spain. Crit. Rev. Biotechnol. 42: 201–219.
Andow, D. A. 2008. The risk of resistance evolution in insects to transgenic
insecticidal crops. Collect. Biosaf. Rev. 4: 142–199.
Andow, D. A., and D. N. Alstad. 1998. F2 screen for rare resistance alleles. J.
Econ. Entomol. 91: 572–578.
Andow, D. A., and A. R. Ives. 2002. Monitoring and adaptive resistance man-
agement. Ecol. Appl. 12: 1378–1390.
Andreadis, S. S., F. Álvarez-Alfageme, I. Sánchez-Ramos, T. J. Stodola, D.
A. Andow, P. G. Milonas, M. Savopoulou-Soultani, and P. Castañera.
2007. Frequency of resistance to Bacillus thuringiensis toxin Cry1Ab in
Greek and Spanish population of Sesamia nonagrioides (Lepidoptera:
Noctuidae). J. Econ. Entomol. 100: 195–201.
Andreadis, S. S., Z. Vryzas, E. Papadopoulou-Mourkidou, and M. Savopoulou-
Soultani. 2008. Age-dependent changes in tolerance to cold and accumula-
Downloaded from https://academic.oup.com/jee/article/116/2/275/6968981 by guest on 30 April 2023
tion of cryoprotectants in overwintering and non-overwintering larvae of
European corn borer Ostrinia nubilalis. Physiol. Entomol. 33: 365–371.
Anglade, P. 1972. Les Sesamie, pp. 1389–1401. In A. S. Balachowski (ed.),
Entomologie appliquée à L’agriculture, vol. II, Lèpidoptères. Masson et
Cie, Paris.
Archer, T., G. Schuster, C. Patrick, G. Cronholm, E. Bynum, Jr, and W.
Morrison. 2000. Whorl and stalk damage by European and Southwestern
corn borers to four events of Bacillus thuringiensis transgenic maize. Crop
Prot. 19: 181–190.
Areal, F. J., and L. Riesgo. 2022. Sustainability of Bt maize in Spain (1998-
2021): an economic, social and environmental analysis. Fundación
Antama, Madrid, Spain.
Arias-Martín, M., M. Haidukowski, G. P. Farinós, and B. Patiño. 2021. Role
of Sesamia nonagrioides and Ostrinia nubilalis as vectors of Fusarium spp.
and contribution of corn borer-resistant Bt maize to mycotoxin reduction.
Toxins. 13: 780.
Badosa, E., C. Moreno, and E. Montesinos. 2004. Lack of detection of am-
picillin resistance gene transfer from Bt176 transgenic corn to culturable
bacteria under field conditions. FEMS Microbiol. Ecol. 48: 169–178.
Bailey, R. I., D. Bourguet, A. L. E. Pallec, and S. Ponsard. 2007. Dispersal pro-
pensity and settling preferences of European corn borers in maize field
borders. J. Appl. Ecol. 44: 385–394.
Bates, S. L., J. Zhao, R. T. Roush, and A. M. Shelton. 2005. Insect resistance
management in GM crops: past, present and future. Nat. Biotechnol. 23:
57–62.
Bažok, R., D. Lemić, F. Chiarini, and L. Furlan. 2021. Western corn rootworm
(Diabrotica virgifera virgifera LeConte) in Europe: current status and sus-
tainable pest management. Insects. 12: 195.
Bertho, L., K. Schmidt, J. Schmidtke, I. Brants, R. F. Cantón, C. Novillo, and
G. Head. 2020. Results from ten years of post-market environmental
monitoring of genetically modified MON 810 maize in the European
Union. PLoS One. 15: e0217272.
Bethenod, M., Y. Thomas, F. Rousset, B. Frérot, L. Pélozuelo, G. Genestier, and
D. Bourguet. 2005. Genetic isolation between two sympatric host plant
races of the European corn borer, Ostrinia nubilalis Hübner. II: assortative
mating and host-plant preferences for oviposition. Heredity. 94: 264–270.
Bohnenblust, E. W., J. A. Breining, J. A. Shaffer, S. J. Fleischer, G. W. Roth, and
J. F. Tooker. 2014. Current European corn borer, Ostrinia nubilalis, injury
levels in the northeastern United States and the value of Bt field corn. Pest
Manag. Sci. 70: 1711–1719.
Bourguet, D., M. T. Bethenod, N. Pasteur, and F. Viard. 2000a. Gene flow
in the European corn borer Ostrinia nubilalis: implications for the sus-
tainability of transgenic insecticidal maize. Proc. R. Soc. B Biol. Sci. 267:
117–122.
Bourguet, D., M. T. Bethenod, C. Trouve, and F. Viard. 2000b. Host–plant
diversity of the European corn borer Ostrinia nubilalis: what value for
sustainable transgenic insecticidal Bt maize? Proc. R. Soc. B: Biol. Sci. 267:
1177–1184.
Bourguet, D., A. Genissel, and M. Raymond. 2000c. Insecticide resistance and
dominance levels. J. Econ. Entomol. 93: 1588–1595.
Bradshaw, C. J., B. Leroy, C. Bellard, D. Roiz, C. Albert, A. Fournier, M.
Barbet-Massin, J. Salles, F. Simard, and F. Courchamp. 2016. Massive yet
Journal of Economic Entomology, 2023, Vol. 116, No. 2
grossly underestimated global costs of invasive insects. Nat. Commun. 7:
1–8.
Brookes, G. 2019. Twenty-one years of using insect resistant (GM) maize in
Spain and Portugal: farm-level economic and environmental contributions.
GM Crops Food. 10: 90–101.
Buntin, G. D., J. N. All, R. D. Lee, and D. M. Wilson. 2004. Plant-incorporated
Bacillus thuringiensis resistance for control of fall armyworm and
corn earworm (Lepidoptera: Noctuidae) in corn. J. Econ. Entomol. 97:
1603–1611.
Butrón, A., R. Malvar, M. Cartea, A. Ordás, and P. Velasco. 1999. Resistance
of maize inbreds to pink stem borer. Crop Sci. 39: 102–107.
Camargo, A. M., D. A. Andow, P. Castañera, and G. P. Farinós. 2018. First de-
tection of a Sesamia nonagrioides resistance allele to Bt maize in Europe.
Sci. Rep. 8: 1–7.
Camargo, A. M., M. Arias-Martín, P. Castañera, and G. P. Farinós. 2020.
Performance of Sesamia nonagrioides on cultivated and wild host plants:
Implications for Bt maize resistance management. Pest Manag. Sci. 76:
3657–3666.
Campagne, P., M. Kruger, R. Pasquet, B. Le Ru, and J. Van den Berg. 2013.
Dominant inheritance of field-evolved resistance to Bt corn in Busseola
fusca. PLoS One. 8: e69675.
Carrière, Y., C. Ellers-Kirk, M. Sisterson, L. Antilla, M. Whitlow, T. J. Dennehy,
and B. E. Tabashnik. 2003. Long-term regional suppression of pink boll-
worm by Bacillus thuringiensis cotton. Proc. Natl. Acad. Sci. U.S.A. 100:
1519–1523.
Carrière, Y., D. W. Crowder, and B. E. Tabashnik. 2010. Evolutionary ecology
of insect adaptation to Bt crops. Evol. Appl. 3: 561–573.
Carrière, Y., N. Crickmore, and B. E. Tabashnik. 2015. Optimizing pyramided
transgenic Bt crops for sustainable pest management. Nat. Biotechnol. 33:
161–168.
Carrière, Y., J. A. Fabrick, and B. E. Tabashnik. 2016. Can pyramids and seed
mixtures delay resistance to Bt crops? Trends Biotechnol. 34: 291–302.
Carrière, Y., Z. Brown, S. Aglasan, P. Dutilleul, M. Carroll, G. Head, B. E.
Tabashnik, P. Søgaard Jørgensen, and S. P. Carroll. 2020. Crop rotation
mitigates impacts of corn rootworm resistance to transgenic Bt corn. Proc.
Natl. Acad. Sci. U.S.A. 117: 18385–18392.
Castañera, P. 1986. Plagas del maíz. IV Jornadas Técnicas sobre el Maíz,
Lérida. Plagas: pp. 1–24. Ministerio de Agricultura Pesca y Alimentación,
Madrid, Spain.
Castañera, P., G. P. Farinós, F. Ortego, and D. A. Andow. 2016. Sixteen years
of Bt maize in the EU hotspot: why has resistance not evolved? PLoS One.
11: e0154200.
Chandrasena, D. I., A. M. Signorini, G. Abratti, N. P. Storer, M. L. Olaciregui,
A. P. Alvesd, and C. D. Pilcherd. 2017. Characterization of field-evolved re-
sistance to Bacillus thuringiensis-derived Cry1F δ-endotoxin in Spodoptera
frugiperda populations from Argentina. Pest Manag. Sci. 74: 746–754.
CSCV (Centro de Sanidad y Certificación Vegetal) 2021. Información
Fitosanitaria 2021. https://www.aragon.es/documents/20127/77520647/
Informacion_Fitosanitaria_202108.pdf/b97398d7-571c-ba96-c543-2ac7
1e96e0da?t=1627899511563.
Dalecky, A., S. Ponsard, R. I. Bailey, C. Pélissier, and D. Bourguet. 2006.
Resistance evolution to Bt crops: predispersal mating of European corn
borers. PLoS One. 4: e181.
De La Poza, M., G. P. Farinós, B. Beroiz, F. Ortego, P. Hernández-Crespo, and
P. Castañera. 2008. Genetic structure of Sesamia nonagrioides (Lefebvre)
populations in the Mediterranean area. Environ. Entomol. 37: 1354–1360.
Devos, Y., D. Reheul, D. De Waele, and L. Van Speybroeck. 2006. The interplay
between societal concerns and the regulatory frame on GM crops in the
European Union. Environ. biosaf. Res. 5: 127–149.
DGAV (Direção-Geral de Alimentação e Veterinária) 2022. Regras para o
cultivo de plantas geneticamente modificadas. https://www.dgav.pt/plantas/
conteudo/autorizacoes-de-cultivo/variedades-geneticamente-modificadas/
regras-para-o-cultivo-de-plantas-geneticamente-modificadas/#.
Dively, G. P., P. D. Venugopal, and C. Finkenbinder. 2016. Field-evolved resist-
ance in corn earworm to Cry proteins expressed by transgenic sweet corn.
PLoS One. 11: e0169115.
Dively, G. P., P. D. Venugopal, D. Bean, J. Whalen, K. Holmstrom, T. P. Kuhar,
H. B. Doughty, T. Patton, W. Cissel, and W. D. Hutchison. 2018. Regional
285
pest suppression associated with widespread Bt maize adoption benefits
vegetable growers. Proc. Natl. Acad. Sci. U.S.A. 115: 3320–3325.
Dorhout, D. L., T. W. Sappington, and M. E. Rice. 2008. Evidence for obligate
migratory flight behavior in young European corn borer (Lepidoptera:
Crambidae) females. Environ. Entomol. 37: 1280–1290.
Edde, P. A. 2022. 7 - Arthropod pests of maize Zea mays (L.), pp. 410–465.
In P. A. Edde (ed.), Field crop arthropod pests of economic importance.
Academic Press, Richmond, VA.
EFSA (European Food Safety Authority). 2015. Clarifications on EFSA GMO
Panel recommendations on the Insect Resistance Management plan for
genetically modified maize MON 810. EFSA Supporting Publication. 12:
EN-842.
EFSA (European Food Safety Authority). 2016a. Scientific opinion on the
annual post-market environmental monitoring (PMEM) report from
Monsanto Europe S.A. on the cultivation of genetically modified maize
MON 810 in 2014. EFSA J. 15: 4805.
Downloaded from https://academic.oup.com/jee/article/116/2/275/6968981 by guest on 30 April 2023
EFSA (European Food Safety Authority). 2016b. Relevance of new scientific
evidence on the occurrence of teosinte in maize fields in Spain and France
for previous environmental risk assessment conclusions and risk man-
agement recommendations on the cultivation of maize events MON810,
Bt11, 1507 and GA21. EFSA Supporting Publication. 2016: EN-1094.
EFSA (European Food Safety Authority). 2017. Scientific opinion on the
annual post-market environmental monitoring (PMEM) report from
Monsanto Europe S.A. on the cultivation of genetically modified maize
MON 810 in 2015. EFSA J. 15: 4805.
EFSA (European Food Safety Authority). 2018. Scientific opinion on the
annual post-market environmental monitoring (PMEM) report from
Monsanto Europe S.A. on the cultivation of genetically modified maize
MON 810 in 2016. EFSA J. 16: 4805.
EFSA (European Food Safety Authority). 2022. Statement on the update
of environmental risk assessment conclusions and risk management
recommendations of EFSA (2016) on EU teosinte. EFSA J. 20: 7228.
Eizaguirre, M., and A. A. Fantinou. 2012. Abundance of Sesamia nonagrioides
(Lef.) (Lepidoptera: Noctuidae) on the edges of the Mediterranean Basin.
Psyche. 2012: 1–7.
Eizaguirre, M., C. López, and A. Sans. 2002. Maize phenology influences field
diapause induction of Sesamia nonagrioides (Lepidoptera: Noctuidae).
Bull. Entomol. Res. 92: 439–443.
Eizaguirre, M., C. Lopez, and R. Albajes. 2004. Dispersal capacity in the Mediterranean
corn borer, Sesamia nonagrioides. Entomol. Exp. Appl. 113: 25–34.
Eizaguirre, M., R. Albajes, C. Lopez, J. Eras, B. Lumbierres, and X. Pons. 2006.
Six years after the commercial introduction of Bt maize in Spain: field eval-
uation, impact and future prospects. Transgenic Res. 15: 1–12.
Eizaguirre, M., F. Madeira, and C. López. 2010. Effects of Bt maize on non-
target lepidopteran pests. IOBC/WPRS Bull. 52: 49–55.
Engels, H., D. Bourguet, L. Cagáň, B. Manachini, I. Schuphan, T. J. Stodola, A.
Micoud, C. Brazier, C. Mottet, and D. A. Andow. 2010. Evaluating resist-
ance to Bt toxin Cry1Ab by F2 screen in European populations of Ostrinia
nubilalis (Lepidoptera: Crambidae). J. Econ. Entomol. 103: 1803–1809.
EPPO (European and Mediterranean Plant Protection Organization). 2021.
First report of Spodoptera frugiperda in the Canary Islands, Spain. EPPO
Reporting Service no. 03 March 2021 (Num. 2021/053).
EPPO (European and Mediterranean Plant Protection Organization). 2022.
Special alert – Fall armyworm, Spodoptera frugiperda, found in the
EPPO region. https://www.eppo.int/ACTIVITIES/plant_quarantine/
shortnotes_qps/fall_armyworm.
Erasmus, R., R. Pieters, H. Du Plessis, A. Hilbeck, M. Trtikova, A. Erasmus,
and J. Van den Berg. 2019. Introgression of a cry1Ab transgene into open
pollinated maize and its effect on Cry protein concentration and target
pest survival. PLoS One. 14: e0226476.
Eriksson, D., E. de Andrade, B. Bohanec, S. Chatzopoulou, R. Defez, N. Leiva
Eriksson, P. van der Meer, B. van der Meulen, A. Ritala, László Sági, et al.
2019. Implementing an EU opt-in mechanism for GM crop cultivation.
EMBO Rep. 20: e48036.
EuropaBio. 2017. Harmonised insect resistance management (IRM) plan for
cultivation of Bt maize (single insecticidal trait) in the EU. https://food.
ec.europa.eu/system/files/2017-10/gmo_rep-stud_mon-810_report-2016_
app-06.pdf.
286
European Commission. 2001. Directive 2001/18/EC on the deliberate release
into the environment of genetically modified organisms and repealing
Council Directive 90/220/EC. EC Off. J. Eur. Commun. L106: 1–38.
European Commission. 2010. Commission recommendation of 13 July 2010
on guidelines for the development of national co-existence measures to
avoid the unintended presence of GMOs in conventional and organic
crops. Off. J. Eur. Union: EC2010/C 200/01.
European Commission. 2022a. Health and Food Safety-Genetically Modified
Organisms-Register. https://webgate.ec.europa.eu/dyna/gm_register/
index_en.cfm.
European Commission. 2022b. Monitoring plans and reports. https://food.
ec.europa.eu/plants/genetically-modified-organisms/post-authorisation/
monitoring-plans-and-reports_en#pmem-reports-for-gm-food-and-feed.
EUROPHYT (European Union Notification System for Plant Health
Interceptions). 2022. Interceptions annual reports. https://food.
ec.europa.eu/plants/plant-health-and-biosecurity/europhyt/
interceptions-annual-reports_en.
Farinós, G. P., M. De La Poza, P. Hernández-Crespo, F. Ortego, and P.
Castañera. 2004. Resistance monitoring of field populations of the corn
borers Sesamia nonagrioides and Ostrinia nubilalis after 5 years of Bt
maize cultivation in Spain. Entomol. Exp. Appl. 110: 23–30.
Farinós, G. P., S. S. Andreadis, M. De la Poza, G. K. Mironidis, F. Ortego, M.
Savopoulou-Soultani, and P. Castañera. 2011. Comparative assessment of
the field-susceptibility of Sesamia nonagrioides to the Cry1Ab toxin in
areas with different adoption rates of Bt maize and in Bt-free areas. Crop
Prot. 30: 902–906.
Farinós, G. P., P. Hernández-Crespo, F. Ortego, and P. Castañera. 2018.
Monitoring of Sesamia nonagrioides resistance to MON 810 maize in the
European Union: lessons from a long-term harmonized plan. Pest Manag.
Sci. 74: 557–568.
Fearing, P. L., D. Brown, D. Vlachos, M. Meghji, and L. Privalle. 1997.
Quantitative analysis of CryIA (b) expression in Bt maize plants, tissues,
and silage and stability of expression over successive generations. Mol.
Breed. 3: 169–176.
Figueiredo, D., and J. Araújo. 1996. Factores de mortalidade de Sesamia
nonagrioides Lef. (Lepidoptera: Noctuidae) em Portugal. I-Parasitóides.
Bol. Sanid. Veg. Plagas. 22: 251–260.
Galichet, P. 1982. Hibernation d’une population de Sesamia nonagrioides Lef.
(Lep. Noctuidae) en France meridionale. Agronomie. 2: 561–566.
Gaspers, C. 2009. The European corn borer (Ostrinia nubilalis, Hbn.), its sus-
ceptibility to the Bt-toxin Cry1F, its pheromone races and its gene flow in
Europe in view of an insect resistance management. PhD Thesis, RWTH
Aachen University, RWTH Publications, Germany.
Gassmann, A. J. 2021. Resistance to Bt maize by western corn rootworm:
effects of pest biology, the pest–crop interaction and the agricultural land-
scape on resistance. Insects. 12: 136.
Gassmann, A. J., J. L. Petzold-Maxwell, R. S. Keweshan, and M. W. Dunbar.
2011. Field-evolved resistance to Bt maize by western corn rootworm.
PLoS One. 6: e22629.
GENCAT (Generalitat de Catalunya). 2022. Registre d’OMG de Catalunya
(ROMGCat). http://agricultura.gencat.cat/ca/departament/estadistiques/
agricultura/estadistiques-omg/.
Gilioli, G., G. Sperandio, A. Simonetto, M. Ciampitti, and P. Gervasio. 2022.
Assessing the risk of establishment and transient populations of Spodoptera
frugiperda in Europe. J. Pest Sci. doi:10.1007/s10340-022-01517-0.
Goergen, G., P. L. Kumar, S. B. Sankung, A. Togola, and M. Tamò. 2016.
First report of outbreaks of the fall armyworm Spodoptera frugiperda (JE
Smith) (Lepidoptera, Noctuidae), a new alien invasive pest in West and
Central Africa. PLoS One. 11: e0165632.
Gomez-Barbero, M., J. Berbel, and E. Rodriguez-Cerezo. 2008. Bt corn in
Spain: the performance of the EU’s first GM crop. Nat. Biotechnol. 26:
384–386.
González-Cabrera, J., G. P. Farinós, S. Caccia, M. Díaz-Mendoza, P. Castañera,
M. G. Leonardi, B. Giordana, and J. Ferré. 2006. Toxicity and mode of ac-
tion of Bacillus thuringiensis Cry proteins in the Mediterranean corn borer,
Sesamia nonagrioides (Lefebvre). Appl. Environ. Microbiol. 72: 2594–2600.
González-Cabrera, J., M. García, P. Hernández-Crespo, G. P. Farinós, F.
Ortego, and P. Castañera. 2013. Resistance to Bt maize in Mythimna
Journal of Economic Entomology, 2023, Vol. 116, No. 2
unipuncta (Lepidoptera: Noctuidae) is mediated by alteration in Cry1Ab
protein activation. Insect Biochem. Mol. Biol. 43: 635–643.
González-Núñez, M., F. Ortego, and P. Castañera. 2000. Susceptibility of
Spanish populations of the corn borers Sesamia nonagrioides (Lepidoptera:
Noctuidae) and Ostrinia nubilalis (Lepidoptera: Crambidae) to a Bacillus
thuringiensis endotoxin. J. Econ. Entomol. 93: 459–463.
Gould, F. 1998. Sustainability of transgenic insecticidal cultivars: integrating
pest genetics and ecology. Annu. Rev. Entomol. 43: 701–726.
Grimi, D. A., B. Parody, M. L. Ramos, M. Machado, F. Ocampo, A. Willse,
S. Martinelli, and G. Head. 2018. Field-evolved resistance to Bt maize in
sugarcane borer (Diatraea saccharalis) in Argentina. Pest Manag. Sci. 74:
905–913.
Head, G. P., and J. Greenplate. 2012. The design and implementation of in-
sect resistance management programs for Bt crops. GM Crops Food. 3:
144–153.
Hilal, A. 1977. Mise en evidence d’un etat de diapause vraie chez Sesamia
Downloaded from https://academic.oup.com/jee/article/116/2/275/6968981 by guest on 30 April 2023
nonagrioides Lef. (Lepidoptera, Noctuidae). C. R. Acad. Sci. Ser. D. Paris.
285: 365.
Hu, Y., and D. A. Andow. 2011. Field observations of Ostrinia nubilalis eclo-
sion and post-eclosion activity of females around their natal plants. Insect
Sci. 18: 712–718.
Huang, F. 2021. Dominance and fitness costs of insect resistance to genetically
modified Bacillus thuringiensis crops. GM Crops Food. 12: 192–211.
Hundleby, P. A., and W. A. Harwood. 2019. Impacts of the EU GMO regula-
tory framework for plant genome editing. Food Energy Secur. 8: e00161.
Hutchison, W. D., E. Burkness, P. Mitchell, R. Moon, T. Leslie, S. J. Fleischer,
M. Abrahamson, K. Hamilton, K. Steffey, and M. Gray. 2010. Areawide
suppression of European corn borer with Bt maize reaps savings to non-Bt
maize growers. Science. 330: 222–225.
ISAAA (International Service for the Acquisition of Agri-Biotech Applications).
2019. Global status of commercialized biotech/GM crops, vol. 55. ISAAA
Brief, Ithaca, NY.
ISAAA (International Service for the Acquisition of Agri-Biotech
Applications). 2022. GM approval database. https://www.isaaa.org/
gmapprovaldatabase/.
Kathage, J., M. Gómez-Barbero, and E. Rodríguez-Cerezo. 2016. Framework
for assessing the socio-economic impacts of Bt maize cultivation (No.
JRC103197). JRC Reports, European Commission. Joint Research Centre,
Seville, Spain.
Keszthelyi, S. 2010. Flight dynamics analysis of the European corn borer
(Ostrinia nubilalis (Hübner)) populations in Hungary from the second
part of the twentieth century until the present. Arch. Phytopathol.
Pflanzenschutz. 43: 1286–1294.
Kfir, R., W. Overholt, Z. Khan, and A. Polaszek. 2002. Biology and manage-
ment of economically important lepidopteran cereal stem borers in Africa.
Annu. Rev. Entomol. 47: 701–731.
Le Corre, V., M. Siol, Y. Vigouroux, M. I. Tenaillon, and C. Délye. 2020.
Adaptive introgression from maize has facilitated the establishment
of teosinte as a noxious weed in Europe. Proc. Nat. Acad. Sci. 117:
25618–25627.
LeOra Software. 2002-2022. POLO PlusV1.0, Probit and Logit Analysis.
LeOra Software Co., Petaluma, CA, USA.
Le Rü, B. P., G. O. Ong’amo, P. Moyal, E. Muchugu, L. Ngala, B. Musyoka,
Z. Abdullah, T. Matama-Kauma, V. Y. Lada, and B. Pallangyo. 2006.
Geographic distribution and host plant ranges of East African noctuid
stem borers. Ann. Soc. Entomol. Fr. 42: 353–361.
Levine, E., J. L. Spencer, S. A. Isard, D. W. Onstad, and M. E. Gray. 2002.
Adaptation of the western corn rootworm to crop rotation: evolution of
a new strain in response to a management practice. Am. Entomol. 48:
94–107.
Logrieco, A., G. Mule, A. Moretti, and A. Bottalico. 2002. Toxigenic
Fusarium species and mycotoxins associated with maize ear rot
in Europe, pp. 597–609. In Mycotoxins in plant disease. Springer,
Dordrecht.
Lohn, A. F., M. Trtikova, I. Chapela, R. Binimelis, and A. Hilbeck. 2021.
Transgene behavior in genetically modified teosinte hybrid plants:
transcriptome expression, insecticidal protein production and bioactivity
against a target insect pest. Environ. Sci. Eur. 33: 1–13.
Journal of Economic Entomology, 2023, Vol. 116, No. 2
López, C., A. Sans, and M. Eizaguirre. 1999. Influencia de la planta de maíz
en el apareamiento de Sesamia nonagrioides Lefèbvre (Lepidoptera:
Noctuidae). Invest. Agrar. Produc. Protec. Veg. 14: 415–422.
López, C., A. Sans, L. Asin, and M. EizaGuirre. 2001. Phenological model for
Sesamia nonagrioides (Lepidoptera: Noctuidae). Environ. Entomol. 30:
23–30.
López, C., M. Eizaguirre, and R. Albajes. 2003. Courtship and mating
behaviour of the Mediterranean corn borer, Sesamia nonagrioides
(Lepidoptera: Noctuidae). Span. J. Agric. Res. 1: 43–51.
Magg, T., A. Melchinger, D. Klein, and M. Bohn. 2001. Comparison of Bt
maize hybrids with their non-transgenic counterparts and commercial
varieties for resistance to European corn borer and for agronomic traits.
Plant Breed. 120: 397–403.
Manosathiyadevan, M., V. Bhuvaneshwari, and R. Latha. 2017. Impact of
insects and pests in loss of crop production: a review, pp. 57–67. In A.
Dhanarajan (ed.), Sustainable Agriculture towards Food Security. Springer,
Singapore.
MAPA (Ministerio de Agricultura, Pesca y Alimentación). 2015. Guía de
gestión Integrada de plagas: Maíz. Ministerio de Agricultura, Pesca y
Alimentación, Madrid, Spain.
MAPA (Ministerio de Agricultura, Pesca y Alimentación). 2022a. Registro
Público de Organismos Modificados Geneticamente (OMG). Ministry of
Agriculture, Fisheries and Food. Public Registry of Genetically Modified
Organisms (GMO). https://www.mapa.gob.es/es/agricultura/temas/
biotecnologia/omg/registro-publico-omg/default.aspx.
MAPA (Ministerio de Agricultura, Pesca y Alimentación). 2022b.
Avances de superficies y producciones de cultivos. https://www.
mapa.gob.es/es/estadistica/temas/estadisticas-agrarias/agricultura/
avances-superficies-producciones-agricolas/.
Marçon, P. C., D. B. Taylor, C. E. Mason, R. L. Hellmich, and B. D. Siegfried.
1999a. Genetic similarity among pheromone and voltinism races of
Ostrinia nubilalis (Hubner) (Lepidoptera: Crambidae). Insect. Mol. Biol.
8: 213–221.
Marçon, P. C., L. J. Young, K. L. Steffey, and B. D. Siegfried. 1999b. Baseline
susceptibility of European corn borer (Lepidoptera: Crambidae) to
Bacillus thuringiensis toxins. J. Econ. Entomol. 92: 279–285.
Marques, L. H., A. C. Santos, B. A. Castro, V. F. Moscardini, J. Rosseto, O.
A. Silva, and J. M. Babcock. 2019. Assessing the efficacy of Bacillus
thuringiensis (Bt) pyramided proteins Cry1F, Cry1A. 105, Cry2Ab2, and
Vip3Aa20 expressed in Bt maize against lepidopteran pests in Brazil. J.
Econ. Entomol. 112: 803–811.
Mason, C., M. Rice, D. Calvin, J. Van Duyn, W. Showers, W. Hutchison, J.
Witkowski, R. Higgins, D. Onstad, and G. Dively. 1996. European corn
borer ecology and management. North Central Regional Extension
Publication 327. Iowa State University, Ames.
Mateu, J., E. Mirasol, and J. Almacelles. 2021. Diabrotica virgifera, Servei de
Sanitat Vegetal del Departament d’Acció Climàtica, Alimentació i Agenda
Rural de la Generalitat de Catalunya, vol. 73. http://agricultura.gencat.
cat/web/.content/ag_agricultura/ag02_sanitat_vegetal/ag02_02_plagues/
documents_plagues/fitxers_estatics/fitxa_073_diabrotica-virgifera.pdf.
Meinke, L. J., D. Souza, and B. D. Siegfried. 2021. The use of insecticides to
manage the western corn rootworm, Diabrotica virgifera virgifera, LeConte:
history, field-evolved resistance, and associated mechanisms. Insects. 12: 112.
Meissle, M., P. Mouron, T. Musa, F. Bigler, X. Pons, V. P. Vasileiadis, S. Otto,
D. Antichi, J. Kiss, Z. Pálinkás, et al. 2010. Pests, pesticide use and alter-
native options in European maize production: Current status and future
prospects. J. Appl. Entomol. 134:357–375.
Melamed-Madjar, V., and S. Tam. 1980. A field survey of changes in the com-
position of corn borer populations in Israel. Phytoparasitica. 8: 201–204.
Mencarelli, M., C. Accinelli, and A. Vicari. 2013. Implications of European
corn borer, Ostrinia nubilalis, infestation in an Aspergillus flavus-
biocontrolled corn agroecosystem. Pest Manag. Sci. 69: 1085–1091.
MITECO (Ministerio para la Transición Ecológica y el Reto Demográfico).
2010. Planes de seguimiento ambiental del cultivo de maíz modificado
genéticamente en España. Ministerio para la Transición Ecológica y el
Reto Demográfico, https://www.miteco.gob.es/es/calidad-y-evaluacion-
ambiental/temas/biotecnologia/PLAN_DE_SEGUIMIENTO_mar%20
2011_tcm30-189498.pdf.
287
Moreno, A., and J. Gastón. 2020. Cinco nuevas especies para la fauna de
España y otras interesantes informaciones lepidopterológicas para España
y Sudán (Insecta: Lepidoptera). SHILAP Soc. Hispano Luso Am. Lepid.
48: 717–731.
Mrganić, M., R. Bažok, K. M. Mikac, H. A. Benítez, and D. Lemic. 2018.
Two decades of invasive western corn rootworm population monitoring
in Croatia. Insects. 9: 160.
Omoto, C., O. Bernardi, E. Salmeron, R. J. Sorgatto, P. M. Dourado, A.
Crivellari, R. A. Carvalho, A. Willse, S. Martinelli, and G. P. Head. 2016.
Field-evolved resistance to Cry1Ab maize by Spodoptera frugiperda in
Brazil. Pest Manag. Sci. 72: 1727–1736.
Ostrem, J. S., Z. Pan, J. L. Flexner, E. Owens, R. Binning, and L. S. Higgins.
2016. Monitoring susceptibility of western bean cutworm (Lepidoptera:
Noctuidae) field populations to Bacillus thuringiensis Cry1F protein. J.
Econ. Entomol. 109: 847–853.
Pardo, G., A. Cirujeda, A. I. Marí, J. Aibar, S. Fuertes, and A. Taberner. 2016.
Downloaded from https://academic.oup.com/jee/article/116/2/275/6968981 by guest on 30 April 2023
El teosinte: descripción, situación actual en el valle del Ebro y resultados
de los primeros ensayos. Vida Rural. 408: 42–48.
Pardo-Lopez, L., M. Soberon, and A. Bravo. 2013. Bacillus thuringiensis in-
secticidal three-domain Cry toxins: mode of action, insect resistance and
consequences for crop protection. FEMS Microbiol. Rev. 37: 3–22.
Park, J., I. McFarlane, R. Phipps, and G. Ceddia. 2011. The impact of the
EU regulatory constraint of transgenic crops on farm income. New
Biotechnol. 28: 396–406.
Pérez-Hedo, M., T. Marques, C. López, R. Albajes, and M. Eizaguirre. 2012.
Determination of the Cry1Ab toxin in Helicoverpa armigera larvae fed
on a diet containing lyophilized Bt leaves. IOBC/WPRS Bull. 73: 75–81.
Pérez-Hedo, M., D. Reiter, C. López, and M. Eizaguirre. 2013. Processing
of the maize Bt toxin in the gut of Mythimna unipuncta caterpillars.
Entomol. Exp. Appl. 148: 56–64.
Pitt, J., M. H. Taniwaki, and M. Cole. 2013. Mycotoxin production in major
crops as influenced by growing, harvesting, storage and processing, with
emphasis on the achievement of Food Safety Objectives. Food Control.
32: 205–215.
van Rensburg, J. B. J. 2007. First report of field resistance by the stem borer,
Busseola fusca (Fuller) to Bt-transgenic maize. J. Plant Soil. 24: 147–151.
Roush, R. T. 1986. Inbreeding depression and laboratory adaptation in Heliothis
virescens (Lepidoptera: Noctuidae). Ann. Entomol. Soc. Am. 79: 583–587.
Roush, R. T. 1994. Managing pests and their resistance to Bacillus
thuringiensis: can transgenic crops be better than sprays? Biocontrol Sci.
Technol. 4: 501–516.
Roush, R. T. 1998. Two–toxin strategies for management of insecticidal trans-
genic crops: can pyramiding succeed where pesticide mixtures have not?
Philos. Trans. R. Soc. Lond. B, Biol. Sci. 353: 1777–1786.
Sanahuja, G., R. Banakar, R. M. Twyman, T. Capell, and P. Christou. 2011.
Bacillus thuringiensis: a century of research, development and commercial
applications. Plant Biotechnol. J. 9: 283–300.
Showers, W. B., R. L. Hellmich, M. E. Derrick-Robinson, and W. H. Hendrix,
III. 2001. Aggregation and dispersal behavior of marked and released
European corn borer (Lepidoptera: Crambidae) adults. Environ. Entomol.
30: 700–710.
Siegfried, B. D., and R. L. Hellmich. 2012. Understanding successful resistance
management: the European corn borer and Bt corn in the United States.
GM Crops Food. 3: 184–193.
Smith, J. L., M. D. Lepping, D. M. Rule, Y. Farhan, and A. W. Schaafsma. 2017.
Evidence for field-evolved resistance of Striacosta albicosta (Lepidoptera:
Noctuidae) to Cry1F Bacillus thuringiensis protein and transgenic corn
hybrids in Ontario, Canada. J. Econ. Entomol. 110: 2217–2228.
Smith, J. L., Y. Farhan, and A. W. Schaafsma. 2019. Practical resistance
of Ostrinia nubilalis (Lepidoptera: Crambidae) to Cry1F Bacillus
thuringiensis maize discovered in Nova Scotia, Canada. Sci. Rep. 9: 1–10.
Storer, N. P., J. M. Babcock, M. Schlenz, T. Meade, G. D. Thompson, J. W.
Bing, and R. M. Huckaba. 2010. Discovery and characterization of field
resistance to Bt maize: Spodoptera frugiperda (Lepidoptera: Noctuidae) in
Puerto Rico. J. Econ. Entomol. 103: 1031–1038.
Székács, A., E. Lauber, J. Juracsek, and B. Darvas. 2010a. Cry1Ab toxin pro-
duction of MON 810 transgenic maize. Environ. Toxicol. Chem. 29:
182–190.
288
Székács, A., E. Lauber, E. Takács, and B. Darvas. 2010b. Detection of Cry1Ab
toxin in the leaves of MON 810 transgenic maize. Anal. Bioanal. Chem.
396: 2203–2211.
Tabashnik, B. E. 1994. Evolution of resistance to Bacillus thuringiensis. Ann.
Rev. Entomol. 39: 79.
Tabashnik, B. E., and Y. Carrière. 2019. Global patterns of resistance to Bt
crops highlighting pink bollworm in the United States, China, and India. J.
Econ. Entomol. 112: 2513–2523.
Tabashnik, B. E., A. J. Gassmann, D. W. Crowder, and Y. Carrière. 2008.
Insect resistance to Bt crops: evidence versus theory. Nat. Biotechnol. 26:
199–202.
Tabashnik, B. E., T. Brévault, and Y. Carrière. 2013. Insect resistance
to Bt crops: lessons from the first billion acres. Nat. Biotechnol. 31:
510–521.
Tabashnik, B. E., J. A. Fabrick and Y. Carrière. 2022. Global patterns of insect
resistance to transgenic Bt crops: The first 25 years. J. Econ. Entomol.,
in press.
Tagliabue, G. 2017. The EU legislation on ‘GMOs’ between nonsense and pro-
tectionism: an ongoing Schumpeterian chain of public choices. GM Crops
Food. 8: 57–73.
Tan, S. Y., B. F. Cayabyab, E. P. Alcantara, F. Huang, K. He, K. W. Nickerson, and
B. D. Siegfried. 2013. Comparative binding of Cry1Ab and Cry1F Bacillus
thuringiensis toxins to brush border membrane proteins from Ostrinia
nubilalis, Ostrinia furnacalis and Diatraea saccharalis (Lepidoptera:
Crambidae) midgut tissue. J. Invertebr. Pathol. 114: 234–240.
Journal of Economic Entomology, 2023, Vol. 116, No. 2
Tepa-Yotto, G. T., H. E. Tonnang, G. Goergen, S. Subramanian, E. Kimathi,
E. M. Abdel-Rahman, D. Flø, K. H. Thunes, K. K. Fiaboe, and S. Niassy.
2021. Global habitat suitability of Spodoptera frugiperda (JE Smith)
(Lepidoptera, Noctuidae): key parasitoids considered for its biological
control. Insects. 12: 273.
Thieme, T., C. Buuk, K. Gloyna, F. Ortego, and G. P. Farinós. 2018. Ten
years of MON 810 resistance monitoring of field populations of Ostrinia
nubilalis in Europe. J. Appl. Entomol. 142: 192–200.
Van den Berg, J., B. Greyvenstein, and H. du Plessis. 2022. Insect resistance
management facing African smallholder farmers under climate change.
Curr. Opin. Insect Sci. 50: 100894.
Van Frankenhuyzen, K. 2009. Insecticidal activity of Bacillus thuringiensis
crystal proteins. J. Invertebr. Pathol. 101: 1–16.
Velasco, P., R. A. Malvar, P. Revilla, A. Butrón, and A. Ordás. 1999. Ear re-
sistance of sweet corn populations to Sesamia nonagrioides (Lepidoptera:
Noctuidae) and Ostrinia nubilalis (Lepidoptera: Pyralidae). J. Econ.
Downloaded from https://academic.oup.com/jee/article/116/2/275/6968981 by guest on 30 April 2023
Entomol. 92: 732–739.
Wright, R. J., M. E. Scharf, L. J. Meinke, X. Zhou, B. D. Siegfried, and L.
D. Chandler. 2000. Larval susceptibility of an insecticide-resistant
western corn rootworm (Coleoptera: Chrysomelidae) population to soil
insecticides: laboratory bioassays, assays of detoxification enzymes, and
field performance. J. Econ. Entomol. 93: 7–13.
Wu, K., Y. Lu, H. Feng, Y. Jiang, and J. Zhao. 2008. Suppression of cotton boll-
worm in multiple crops in China in areas with Bt toxin–containing cotton.
Science. 321: 1676–1678.