Insect Resistance Traits Research Articles

Transgenic Bt Cotton Does Not Disrupt the Top-Down Forces Regulating the Cotton Aphid in Central China.

Top-down force is referred to arthropod pest management delivered by the organisms from higher trophic levels. In the context of prevalent adoption of transgenic Bt crops that produce insecticidal Cry proteins derived from Bacillus thuringiensis (Bt), it still remains elusive whether the top-down forces are affected by the insect-resistant traits that introduced into the Bt crops. We explored how Bt cotton affect the strength of top-down forces via arthropod natural enemies in regulating a non-target pest species, the cotton aphid Aphis gossypii Glover, using a comparative approach (i.e. Bt cotton vs. conventional cotton) under field conditions. To determine top-down forces, we manipulated predation/parasitism exposure of the aphid to their natural enemies using exclusion cages. We found that the aphid population growth was strongly suppressed by the dominant natural enemies including Coccinellids, spiders and Aphidiines parasitoids. Coccinellids, spiders and the assemblage of other arthropod natural enemies (mainly lacewings and Hemipteran bugs) are similarly abundant in both plots, but with the parasitoid mummies less abundant in Bt cotton plots compared to the conventional cotton plots. However, the lower abundance of parasitoids in Bt cotton plots alone did not translate into differential top-down control on A. gossypii populations compared to conventional ones. Overall, the top-down forces were equally strong in both plots. We conclude that transgenic Bt cotton does not disrupt the top-down forces regulating the cotton aphid in central China.

Assessing Utilization and Environmental Risks of Important Genes in Plant Abiotic Stress Tolerance.

Transgenic plants with improved salt and drought stress tolerance have been developed with a large number of abiotic stress-related genes. Among these, the most extensively used genes are the glycine betaine biosynthetic codA, the DREB transcription factors, and vacuolar membrane Na+/H+ antiporters. The use of codA, DREBs, and Na+/H+ antiporters in transgenic plants has conferred stress tolerance and improved plant phenotype. However, the future deployment and commercialization of these plants depend on their safety to the environment. Addressing environmental risk assessment is challenging since mechanisms governing abiotic stress tolerance are much more complex than that of insect resistance and herbicide tolerance traits, which have been considered to date. Therefore, questions arise, whether abiotic stress tolerance genes need additional considerations and new measurements in risk assessment and, whether these genes would have effects on weediness and invasiveness potential of transgenic plants? While considering these concerns, the environmental risk assessment of abiotic stress tolerance genes would need to focus on the magnitude of stress tolerance, plant phenotype and characteristics of the potential receiving environment. In the present review, we discuss environmental concerns and likelihood of concerns associated with the use of abiotic stress tolerance genes. Based on our analysis, we conclude that the uses of these genes in domesticated crop plants are safe for the environment. Risk assessment, however, should be carefully conducted on biofeedstocks and perennial plants taking into account plant phenotype and the potential receiving environment.

Suitability of Soybean Meal from Insect-Resistant Soybeans for Broiler Chickens.

Benning(M) and Benning(MGH) are near-isogenic lines (NILs) of the soybean cultivar Benning, which contain insect-resistance quantitative trait loci (QTLs) from the soybean accession PI 229358. Benning(M) contains QTL-M, which confers antibiosis and antixenosis. In addition to QTL-M, Benning(MGH) contains QTL-G, which confers antibiosis, and QTL-H, which confers antixenosis. Soybean meal was produced from Benning and the NILs. Nutritional composition, digestible amino acid content, and nitrogen-corrected true metabolizable energy (TMEN) were equivalent among soybean meals. A 21-day broiler feeding trial was carried out to determine if the QTLs affect soybean meal quality. Weight gain and feed-to-gain ratio were evaluated. No biologically significant differences were detected for broilers fed Benning, Benning(M), and Benning(MGH). This demonstrates that soybean meal produced from the insect-resistant NILs is equivalent to soybean meal produced from their non-insect-resistant parent cultivar for broiler weight gain.

Stacking of Herbicide-Tolerant and Insect-Resistance Traits in Seeds Has Increased

Stacking of Herbicide-Tolerant and Insect-Resistance Traits in Seeds Has Increased

Development of certified matrix-based reference material of genetically modified rice event TT51-1 for real-time PCR quantification.

In 2009, the genetically modified (GM) rice event TT51-1 with an engineered insect resistance trait became the first GM rice event to be granted certification for safe production in China, and its derivative lines Bt 63 and Huahui No.1 are expected to be commercialized soon. The development of certified reference material (CRM) for TT51-1 is necessary to monitor and inspect the TT51-1 event and its derivates. In this work, we developed four matrix-based TT51-1 rice CRMs (TT51-1a, TT51-1b, TT51-1c, and TT51-1d) with different TT51-1 mass fraction ratios by blending seed powders of homozygous TT51-1 and its recipient cultivar Minghui 63. The between-bottle homogeneity and the within-bottle homogeneity were tested, and good results were obtained. The potential degradation during transportation and shelf life were evaluated, and demonstrated an expiration period of at least 36 months. The characterization values of the four TT51-1 CRMs based on the mass fraction ratio were 1000.000 ± 51.430 g/kg, 49.940 ± 4.620 g/kg, 9.990 ± 1.110 g/kg, and 4.990 ± 0.620 g/kg, respectively. The characterization values based on the copy number ratio were certified by digital PCR analysis as 97.442 ± 5.253 %, 4.851 ± 0.486 %, 1.042 ± 0.135 %, and 0.556 ± 0.073 %, respectively. These results suggested that the TT51-1 matrix-based CRMs developed are of high quality and can be used as potential calibrators for TT51-1 GM rice inspection and monitoring.

Do Sesamia nonagrioides (Lepidoptera; Noctuidae) gravid females discriminate between Bt or multivitamin corn varieties? Role of olfactory and visual cues.

The Mediterranean corn borer, Sesamia nonagrioides Lefèbvre, is a key pest of corn and a main target of Bacillus thuringiensis (Bt) corn in Northeast Spain. Trends for future biotechnology crops indicate that Bt, non-Bt, and stacked corn varieties with metabolic pathways for vitamin-increased traits could coexist in same region. Knowledge of the oviposition response of gravid females of S. nonagrioides to these different varieties could be extremely important for managing strategies aimed for delaying resistance development. In dual-choice assays, we examined the host preference of gravid females of S. nonagrioides for four corn varieties: a new transgenic corn with increased vitamin levels, its near isogenic counterpart (M37W), a Bt corn plant, and its near isogenic counterpart. Olfactory cues were the predominant ones when gravid females looked for a suitable host to lay eggs, and no synergistic effects were observed when both visual and olfactory cues were present. When the plant was visible, the females preferred the odors emitted by the nontransgenic to its multivitamin transgenic counterpart and when they only could detect the volatiles they also preferred the nontransgenic M37W variety to the Bt corn variety. If gravid females are less attracted to corn with an increased level of vitamins, this could impact insect resistance management and the value of refuge plants, if such traits are stacked with an insect resistance trait.

Structural and biophysical characterization of Bacillus thuringiensis insecticidal proteins Cry34Ab1 and Cry35Ab1.

Bacillus thuringiensis strains are well known for the production of insecticidal proteins upon sporulation and these proteins are deposited in parasporal crystalline inclusions. The majority of these insect-specific toxins exhibit three domains in the mature toxin sequence. However, other Cry toxins are structurally and evolutionarily unrelated to this three-domain family and little is known of their three dimensional structures, limiting our understanding of their mechanisms of action and our ability to engineer the proteins to enhance their function. Among the non-three domain Cry toxins, the Cry34Ab1 and Cry35Ab1 proteins from B. thuringiensis strain PS149B1 are required to act together to produce toxicity to the western corn rootworm (WCR) Diabrotica virgifera virgifera Le Conte via a pore forming mechanism of action. Cry34Ab1 is a protein of ∼14 kDa with features of the aegerolysin family (Pfam06355) of proteins that have known membrane disrupting activity, while Cry35Ab1 is a ∼44 kDa member of the toxin_10 family (Pfam05431) that includes other insecticidal proteins such as the binary toxin BinA/BinB. The Cry34Ab1/Cry35Ab1 proteins represent an important seed trait technology having been developed as insect resistance traits in commercialized corn hybrids for control of WCR. The structures of Cry34Ab1 and Cry35Ab1 have been elucidated to 2.15 Å and 1.80 Å resolution, respectively. The solution structures of the toxins were further studied by small angle X-ray scattering and native electrospray ion mobility mass spectrometry. We present here the first published structure from the aegerolysin protein domain family and the structural comparisons of Cry34Ab1 and Cry35Ab1 with other pore forming toxins.

Efficacy of Pyramided Bt Proteins Cry1F, Cry1A.105, and Cry2Ab2 Expressed in SmartStax Corn Hybrids Against Lepidopteran Insect Pests in the Northern United States

Commercial field corn (Zea mays L.) hybrids transformed to express some or all of the lepidopteran insect-resistant traits present in SmartStax corn hybrids were evaluated for insecticidal efficacy against a wide range oflepidopteran corn pests common to the northern United States, during 2008 to 2011 at locations in 15 states. SmartStax hybrids contain a pyramid of two Bacillus thuringiensis (Bt) derived events for lepidopteran control: event TC1507 expressing Cry1F protein and MON 89034 expressing CrylA.105 + Cry2Ab2. These studies focused on characterization of the relative efficacy of each event when expressed alone or in combination, and compared with non-Bt hybrid. Corn hybrids containing pyramided insecticidal proteins Cry1F + Cry1A.105 + Cry2Ab2 (SmartStax) consistently showed reduced plant feeding damage by a wide range of lepidopteran larvae compared with single event and non-Bt hybrids. Corn hybrids expressing TC1507 or MON 89034 as single or pyramided events were consistently efficacious against Ostrinia nubilalis (Hübner). SmartStax hybrids had less injury from Agrotis ipsilon (Hufnagel) and Striacosta albicosta (Smith) than corn hybrids containing only event MON 89034 but were not more efficacious than single event TC1507 hybrids. Corn hybrids with event MON 89034 provided better control of Helicoverpa zea (Boddie), than event TC1507 alone. Spodoptera frugiperda (J.E. Smith) efficacy was higher for hybrids with pyramid events and single events compared with the non-Bt hybrids. The spectra of activity of events TC1507 and MON 89034 differed. The combination of TC1507 + MON 89034 provided redundant control of some pests where the spectra overlapped and thereby are expected to confer a resistance management benefit.

Insecticidal Activity of Bacillus thuringiensis Cry1Bh1 against Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae) and Other Lepidopteran Pests

Bacillus thuringiensis is an important source of insect resistance traits in commercial crops. In an effort to prolong B. thuringiensis trait durability, insect resistance management programs often include combinations of insecticidal proteins that are not cross resistant or have demonstrable differences in their site of action as a means to mitigate the development of resistant insect populations. In this report, we describe the activity spectrum of a novel B. thuringiensis Cry protein, Cry1Bh1, against several lepidopteran pests, including laboratory-selected B. thuringiensis-resistant strains of Ostrinia nubilalis and Heliothis virescens and progeny of field-evolved B. thuringiensis-resistant strains of Plutella xylostella and Spodoptera frugiperda. Cry1Bh1 is active against susceptible and B. thuringiensis-resistant colonies of O. nubilalis, P. xylostella, and H. virescens in laboratory diet-based assays, implying a lack of cross-resistance in these insects. However, Cry1Bh1 is not active against susceptible or Cry1F-resistant S. frugiperda. Further, Cry1Bh1 does not compete with Cry1Fa or Cry1Ab for O. nubilalis midgut brush border membrane binding sites. Cry1Bh1-expressing corn, while not completely resistant to insect damage, provided significantly better leaf protection against Cry1Fa-resistant O. nubilalis than did Cry1Fa-expressing hybrid corn. The lack of cross-resistance with Cry1Ab and Cry1Fa along with independent membrane binding sites in O. nubilalis makes Cry1Bh1 a candidate to further optimize for in-plant resistance to this pest.

The roles of risk and ambiguity in technology adoption

We study the impacts of risk and ambiguity aversion on the adoption of new technologies, specifically genetically modified (GM) corn and soy seeds. We conduct experiments measuring risk and ambiguity aversion with Midwestern grain farmers. Risk aversion has only a small impact on the timing of adoption of GM soy, while ambiguity-aversion has a large impact speeding up farmer adoption of GM corn. We hypothesize that this unusual finding is due to the fact that GM corn often contains an insect-resistance trait which reduces the ambiguity of pest damages for adopters. GM soy never contains this insect-resistance trait. This highlights the importance of distinguishing between risk and ambiguity when studying the effects of aversion to uncertainty on adoption of new technologies.

Simulating evolutionary responses of an introgressed insect resistance trait for ecological effect assessment of transgene flow: a model for supporting informed decision-making in environmental risk assessment.

Predicting outcomes of transgene flow from arable crops requires a system perspective that considers ecological and evolutionary processes within a landscape context. In Europe, the arable weed Raphanus raphanistrum is a potential hybridization partner of oilseed rape, and the two species are ecologically linked through the common herbivores Meligethes spp. Observations in Switzerland show that high densities of Meligethes beetles maintained by oilseed rape crops can lead to considerable damage on R. raphanistrum. We asked how increased insect resistance in R. raphanistrum – as might be acquired through introgression from transgenic oilseed rape – would affect seed production under natural herbivore pressure. In simulation experiments, plants protected against Meligethes beetles produced about twice as many seeds as unprotected plants. All stages in the development of reproductive structures from buds to pods were negatively affected by the herbivore, with the transition from buds to flowers being the most vulnerable. We conclude that resistance to Meligethes beetles could confer a considerable selective advantage upon R. raphanistrum in regions where oilseed rape is widely grown.

Sources of Resistance to the Maize Weevil Sitophilus Zeamais in Tropical Maize

The maize weevil, Sitophilus zeamais (Motschulsky) (Coleoptera: Curculionidae), is among the major storage pests that enhance food insecurity among maize farmers. New sources of resistance to the maize weevil are critical in a successful breeding program to address grain damage by postharvest pests. The objective of the study was to evaluate resistance in maize genotypes to the maize weevil, and consequently their value for use in breeding programs. A total of 175 genotypes, including hybrids, landraces, open-pollinated varieties and checks, were tested for resistance to the maize weevil. The percentage grain damage, weight loss, flour weight and weight of damaged and undamaged grains were measured. Significant differences (P <0.001) were observed among the genotypes for all the traits evaluated. The distribution of the genotypes among the different categories of resistance was an indication of the existence of genetic variation. The most resistant genotypes were CKPH08003 and BRAZ 2451 while the most susceptible were PH 3254 and BRAZ 4, among the hybrids and landraces respectively. Genotypes that were superior to the resistant checks were identified. The percentage weight loss and flour weight were identified as the most important insect-resistance traits for discriminating genotypes as evident from the canonical discriminant analysis. Correlation coefficients among the traits evaluated were highly significant. The resistant hybrids identified can be recommended for release and adoption by farmers, whereas the resistant landraces can act as sources of resistance for use in breeding programs.

Editor's choice: Crop genome plasticity and its relevance to food and feed safety of genetically engineered breeding stacks.

Genetically engineered (GE) stacks, combinations of two or more single transgenic events (i.e. single-locus insertions) that have been produced by crossing sexually compatible parents, are an important and growing sector of the crop seed market. Stacked traits covered 26% of the global transgenic crop area in 2011 and were the fastest growing trait group, with a 31% increase in the area planted compared with 2010 (James, 2011). Stacked traits already dominate the market in some regions. For example, 95% of the cotton (Gossypium spp.) grown in Australia during 2011 had both herbicide tolerance and insect resistance traits (James, 2011). Worldwide, at least 12 countries are now growing stacked varieties, of which nine are developing countries (James, 2011). The rapid adoption of GE stacks has focused attention on whether the safety of such products differs from that of the individual events.

Transgenic insect resistance traits increase corn yield and yield stability

Transgenic insect resistance traits increase corn yield and yield stability

Biotech Crops' Seal of Safety Does Not Convince Skeptics

long-term toxic effects of consuming foods genetically manipulated to promote herbicide tolerance, insect resistance, or other traits, and that longer-term studies are needed to truly demonstrate safety. In 1992, the U.S. Food and Drug Administration called foods containing ingredients derived from the recombinant DNA technology used to make GMOs “substantially similar” to conventional ones and deemed them “generally recognized as safe.” Since then, proponents and government agencies have insisted that the more extensive tests required for drugs or pesticides aren’t necessary. Only a latent trend—such as an inexplicable spike in cancer rates—could signal that consuming GMOs over several decades may lead to health problems stemming from the buildup of toxins in the body.

Weed Management with Glyphosate- and Glufosinate-Based Systems in PHY 485 WRF Cotton

Glyphosate-resistant (GR) Palmer amaranth has become a serious pest in parts of the Cotton Belt. Some GR cotton cultivars also contain the WideStrike™ insect resistance trait, which confers tolerance to glufosinate. Use of glufosinate-based management systems in such cultivars could be an option for managing GR Palmer amaranth. The objective of this study was to evaluate crop tolerance and weed control with glyphosate-based and glufosinate-based systems in PHY 485 WRF cotton. The North Carolina field experiment compared glyphosate and glufosinate alone and in mixtures applied twice before four- to six-leaf cotton. Additional treatments included glyphosate and glufosinate mixed withS-metolachlor or pyrithiobac applied to one- to two-leaf cotton followed by glyphosate or glufosinate alone on four- to six-leaf cotton. All treatments received a residual lay-by application. Excellent weed control was observed from all treatments on most weed species. Glyphosate was more effective than glufosinate on glyphosate-susceptible (GS) Palmer amaranth and annual grasses, while glufosinate was more effective on GR Palmer amaranth. Annual grass and GS Palmer amaranth control by glyphosate plus glufosinate was often less than control by glyphosate alone but similar to or greater than control by glufosinate alone, while mixtures were more effective than either herbicide alone on GR Palmer amaranth. Glufosinate caused minor and transient injury to the crop, but no differences in cotton yield or fiber quality were noted. This research demonstrates glufosinate can be applied early in the season to PHY 485 WRF cotton without concern for significant adverse effects on the crop. Although glufosinate is often less effective than glyphosate on GS Palmer amaranth, GR Palmer amaranth can be controlled with well-timed applications of glufosinate. Use of glufosinate in cultivars with the WideStrike trait could fill a significant void in current weed management programs for GR Palmer amaranth in cotton.

Is the German suspension of MON810 maize cultivation scientifically justified?

We examined the justifications invoked by the German government in April 2009 to suspend the cultivation of the genetically modified maize varieties containing the Bt insect-resistance trait MON810. We have carried out a critical examination of the alleged new data on a potential environmental impact of these varieties, namely two scientific papers describing laboratory force-feeding trials on ladybirds and daphnia, and previous data on Lepidoptera, aquatic and soil organisms. We demonstrate that the suspension is based on an incomplete list of references, ignores the widely admitted case-by-case approach, and confuses potential hazard and proven risk in the scientific procedure of risk assessment. Furthermore, we did not find any justification for this suspension in our extensive survey of the scientific literature regarding possible effects under natural field conditions on non-target animals. The vast majority of the 41 articles published in 2008 and 2009 indicate no impact on these organisms and only two articles indicate a minor effect, which is either inconsistent during the planting season or represents an indirect effect. Publications from 1996 to 2008 (376 publications) and recent meta-analyses do not allow to conclude on consistent effects either. The lower abundance of some insects concerns mainly specialized enemies of the target pest (an expected consequence of its control by Bt maize). On the contrary, Bt maize have generally a lower impact than insecticide treatment. The present review demonstrates that the available meta-knowledge on Cry1Ab expressing maize was ignored by the German government which instead used selected individual studies.Electronic supplementary materialThe online version of this article (doi:10.1007/s11248-009-9297-5) contains supplementary material, which is available to authorized users.

Response to Wilkinson &amp; Tepfer's “Fitness and beyond: preparing for the arrival of GM crops with ecologically important novel characters”

Mike Wilkinson and Mark Tepfer’s commentary (Wilkinson and Tepfer, 2009) is a very welcomed contribution towards more realism in environmental risk assessment (ERA). I share most of their views regarding the next generation of GM crops with ecologically important novel characters. Looking at the past GM crops with herbicide tolerance and insect resistance traits, one must confess that environmental impacts reported so far are – if at all – small compared to other consequences of human activities (e.g. global climatic change or increased salinization of cropland). However, the quantification of ‘fitness’ is not an easy task, as this term clearly depends on the environmental context (e.g. the novel plant in the receiving environment). I know colleagues who recommend measuring ‘plant performance’ instead of ‘fitness’. Then the impact of novel genetically modified characters on the establishment and spread in the context of the receiving environment is a key point. Two considerations will strengthen the regulatory preparation for novel characters: (1) assessing the ecological phenotype and (2) definition of ‘weediness’.

Construction of a BAC Library for a Defoliating Insect-Resistant Soybean and Identification of Candidate Clones Using a Novel Approach

Positional cloning of an insect-resistance quantitative trait locus (QTL) requires the construction of a large-insert genomic DNA library from insect-resistant genotypes. To facilitate cloning of a major defoliating insect-resistance QTL on linkage group M of the soybean genetic map, a bacterial artificial chromosome (BAC) library for PI 229358 was constructed and characterized. The HindIII BAC library contains 55,296 clones with an average insert size 131 kb. This library represents a 6-fold soybean haploid genome equivalents, allowing a 99.8% probability of recovering any specific sequence of interest in soybean. BAC filters were screened with a genomic DNA probe Sat_258sc2 obtained through genome walking from flanking sequences of a simple sequence repeat (SSR) marker, Sat_258, which links to the insect-resistance QTL. Thirteen BAC clones were identified positive for Sat_258sc2, and two of them were confirmed to carry Sat_258. The results suggest that this library is useful in positional cloning of the major insect-resistance QTL, and the approach presented here can be used to screen a BAC library for a SSR marker without requiring the creation of BAC pools.

Divergent Recurrent Selection for Vegetative Phase Change and Effects on Agronomic Traits and Corn Borer Resistance

Vegetative phase change, or the transition from juvenile vegetative to adult vegetative tissues, is essential to the lifecycle of higher plants. In maize (Zea mays L.), juvenile and adult vegetative tissues have distinctly different traits that may confer differing amounts of resistance to disease and insect infestations. Vegetative phase change has been linked to resistance to insects and common rust (Puccinia sorghi Schwein.). Seven cycles of divergent recurrent selection for early and late phase change were evaluated to determine effects of selection on phase change, insect resistance, and agronomic traits. Last leaf with juvenile wax, a trait indicative of the timing of phase change, was the selected trait. Divergent recurrent selection was effective in creating populations that underwent vegetative phase change at distinctly different developmental stages. Last leaf with juvenile wax moved from leaf 8.70 in the original population to leaf 14.48 in the late direction and 6.04 in the early direction. First leaf with adult wax moved from leaf 6.36 in the original population to leaf 8.15 in the late direction and 5.28 in the early direction. Several agronomic traits were also significantly altered by selection including plant and ear height, leaf number, kernel row count, and days to silking. C7Late plants averaged four more leaves and were taller and later flowering than C7Early plants. European corn borer (Ostrinia nubilalis Hubner) feeding damage on the second leaf above the ear was significantly greater in the late phase change direction, and significantly correlated with last leaf with juvenile wax. Most ear traits and European corn borer stalk damage resistance were not altered in a consistent way by selection.