Transgenic Hybrids Research Articles

Histopathological Effects of Bt and TcdA Insecticidal Proteins on the Midgut Epithelium of Western Corn Rootworm Larvae (Diabrotica virgifera virgifera).

Western corn rootworm (WCR, Diabrotica virgifera virgifera LeConte) is a major corn pest in the United States, causing annual losses of over $1 billion. One approach to protect against crop loss by this insect is the use of transgenic corn hybrids expressing one or more crystal (Cry) proteins derived from Bacillus thuringiensis. Cry34Ab1 and Cry35Ab1 together comprise a binary insecticidal toxin with specific activity against WCR. These proteins have been developed as insect resistance traits in commercialized corn hybrids resistant to WCR feeding damage. Cry34/35Ab1 is a pore forming toxin, but the specific effects of Cry34/35Ab1 on WCR cells and tissues have not been well characterized microscopically, and the overall histopathology is poorly understood. Using high-resolution resin-based histopathology methods, the effects of Cry34/35Ab1 as well as Cry3Aa1, Cry6Aa1, and the Photorhabdus toxin complex protein TcdA have been directly visualized and documented. Clear symptoms of intoxication were observed for all insecticidal proteins tested, including swelling and sloughing of enterocytes, constriction of midgut circular muscles, stem cell activation, and obstruction of the midgut lumen. These data demonstrate the effects of these insecticidal proteins on WCR midgut cells, and the collective response of the midgut to intoxication. Taken together, these results advance our understanding of the insect cell biology and pathology of these insecticidal proteins, which should further the field of insect resistance traits and corn rootworm management.

English

Resistance in western corn rootworm to transgenic corn hybrids was first confirmed in 2011 in Midwestern USA, and threatens their continued use. Farmers are often the first line of resistance detection, so their understanding and attitudes toward this issue are critical for improving resistance management. We conducted telephone focus groups during 2013 with farmers who had experienced rootworm resistance. There were four stages in dealing with unexpected rootworm injury: Awareness of a problem, diagnosis, confirmation, and recommendations. Most farmers discovered the problem themselves, but this usually happened too late in the growing season to limit yield loss. Once aware of a problem, farmers first sought help diagnosing the problem from their seed dealer, chemical rep, and/or crop consultant. They considered the problem to be a significant one, both because of its severity and suddenness, and were concerned about their difficulty in obtaining a correct diagnosis. They eventually used extension entomology specialists to confirm the diagnosis. Farmers gathered recommendations from independent consultants, input suppliers, and extension and indicated that they would aggressively deal with the problem, because they were not sure of what would work to protect their crop. They recommended that public extension put more emphasis on increasing awareness of the problem, assessing the extent of the problem and being an unbiased source of information. However, farmers were unlikely to report rootworm injury if the perceived barriers to reporting outweighed the perceived incentives. These barriers were emotional ones, including being unsure who to trust, fear that reporting will be time-consuming, and shame that they did something wrong. The incentive was access to credible advice. They did not automatically acknowledge the broader social benefits of reporting. Thus, extension probably needs to be explicit about these broader benefits to obtain information about the extent of the problem. With the conflicting demands and multiple information sources, it will be a challenge for extension to involve farmers to improve resistance monitoring and management. Key words: Resistance management, focus group, qualitative analysis, Diabrotica virgifera, genetically modified organism, transgenic crop, extension.

Integration of biological control and transgenic insect protection for mitigation of mycotoxins in corn

Biological control is known to be effective in reducing aflatoxin contamination of corn, and some transgenic corn hybrids incur reduced damage from corn earworm (Helicoverpa zea) than comparable non-Bt hybrids. We conducted seven field trials over two years to test the hypothesis that transgenic insect protection and biological control could be integrated to prevent mycotoxin contamination of corn. Corn hybrid N78N3111, expressing the highest degree of transgenic insect protection, was nearly 100 percent free from corn earworm damage and generally had less than half as much contamination from fumonisin compared to N78NGT, a near isogenic corn hybrid without insect protection. This insect protection, however, did not significantly prevent aflatoxin contamination. Soil application of non-aflatoxigenic biocontrol strains of Aspergillus flavus significantly reduced aflatoxin concentrations in corn. Biocontrol strain 21882 of A. flavus was especially effective and reduced aflatoxin contamination by about 90 percent over the seven field trials. There was no significant interaction between the insect protection and biocontrol treatments. Although no synergies were detected, the reduction of mycotoxins by both strategies supports application of both strategies in tandem. Economic factors external to the cost of the technologies will be a major determinant if the mycotoxin mitigation attained by use of these technologies will have a positive economic benefit.

The Arabidopsis ORANGE (AtOR) gene promotes carotenoid accumulation in transgenic corn hybrids derived from parental lines with limited carotenoid pools.

The AtOR gene enhances carotenoid levels in corn by promoting the formation of plastoglobuli when the carotenoid pool is limited, but has no further effect when carotenoids are already abundant. The cauliflower orange (or) gene mutation influences carotenoid accumulation in plants by promoting the transition of proplastids into chromoplasts, thus creating intracellular storage compartments that act as metabolic sink. We overexpressed the Arabidopsis OR gene under the control of the endosperm-specific wheat LMW glutenin promoter in a white corn variety that normally accumulates only trace amounts of carotenoids. The total endosperm carotenoid content in the best-performing AtOR transgenic corn line was 32-fold higher than wild-type controls (~25µg/g DW at 30 days after pollination) but the principal carotenoids remained the same, suggesting that AtOR increases the abundance of existing carotenoids without changing the metabolic composition. We analyzed the expression of endogenous genes representing the carotenoid biosynthesis and MEP pathways, as well as the plastid fusion/translocation factor required for chromoplast formation, but only the DXS1 gene was upregulated in the transgenic corn plants. The line expressing AtOR at the highest level was crossed with four transgenic corn lines expressing different carotenogenic genes and accumulating different carotenoids. The introgression of AtOR increased the carotenoid content of the hybrids when there was a limited carotenoid pool in the parental line, but had no effect when carotenoids were already abundant in the parent. The AtOR gene therefore appears to enhance carotenoid levels by promoting the formation of carotenoid-sequestering plastoglobuli when the carotenoid pool is limited, but has no further effect when carotenoids are already abundant because high levels of carotenoids can induce the formation of carotenoid-sequestering plastoglobuli even in the absence of AtOR.

Pollen-mediated gene flow from transgenic perennial creeping bentgrass and hybridization at the landscape level.

The planting of 162 ha of transgenic glyphosate-resistant creeping bentgrass (Agrostis stolonifera) near Madras, OR, USA, allowed a unique opportunity to study gene flow over time from a perennial outcrossing species at the landscape level. While conducting a four year in situ survey, we collected panicles and leaf tissue samples from creeping bentgrass and its sexually compatible species. Seeds from the panicles were planted, and seedlings were tested in the greenhouse for expression of the transgene. Gene flow via pollen was found in all four years, at frequencies of 0.004 to 2.805%. Chloroplast markers, in combination with internal transcribed spacer nuclear sequence analysis, were used to aid in identification of transgenic interspecific and intergeneric hybrid seedlings found during the testing and of established plants that could not be positively identified in the field. Interspecific transgenic hybrids produced on redtop (Agrostis gigantea) plants in situ were identified three of the four years and one intergeneric transgenic creeping bentgrass x rabbitfoot grass (Polypogon monspeliensis) hybrid was identified in 2005. In addition, we confirmed a non-transgenic creeping bentgrass x redtop hybrid in situ, demonstrating that interspecific hybrids have established in the environment outside production fields. Results of this study should be considered for deregulation of transgenic events, studies of population dynamics, and prediction of gene flow in the environment.

Field-Grown Transgenic Hybrid Poplar with Modified Lignin Biosynthesis to Improve Enzymatic Saccharification Efficiency

Hybrid poplars (Populus nigra L. × Populus maximowiczii A.) were genetically modified through antisense insertion of the 4-coumarate:coenzyme A ligase (4CL) gene. Compositional changes in response to this genetic change were measured in the field after 2 and 3 years of growth. The stem samples were treated with either green liquor or dilute acid pretreatments, representing alkaline and acid pretreatments. The enzymatic saccharification of the untreated and pretreated transgenic poplars were evaluated. After transgenic species were transplanted into the environment, they showed reduced recalcitrance to chemicals (i.e., pretreatments) and enzymes despite their lignin content and S/V ratio being comparable to those of the wild types. Compared to the field-grown poplars, the sugar yield increased up to 103% for untreated transgenic samples and increased 22% for acid- and green liquor-pretreated transgenic samples. This shows that field-grown transgenic hybrid poplars with modified lignin biosynthesis have imp...

Glyphosate tolerant and insect resistant transgenic Bt maize efficacy against shoot borer, cob borer and non-target insect pests

Introducing Bt insect resistance genes into hybrid maize seems to be the most feasible and effective technique accessible to control the pest. Transgenic stack hybrid maize (MON 89034X NK 603) was developed by Monsanto for checking yield losses of maize crop to increase productivity. Transgenic stack hybrid maize was claimed to have both insect protection and herbicide tolerant traits to provide protection to the crop from target pests and also provide effective weed management. MON 89034X NK 603 is 2nd generation glyphosate tolerant with Bt maize technology effective against lepidopteron insect pests with “dual mode of action”. A field experiment was carried out at ICAR-Directorate of Weed Research, Jabalpur for two years during Kharif (rainy season) 2009 and 2010 to evaluate the efficacy of transgenic stack hybrid maize (MON 89034X NK 603) against shoot borer (Chilo partellus), cob borer (Helicoverpa armigera), non-target insect pests and beneficial insects. Treatments consisted of two transgenic stacked hybrids named ‘Hishell’ and ‘‘900M Gold’’ with two conventional hybrids namely ‘Proagro-4640’ and ‘‘HQPM-1’’. Artificial infestation of Chilo partelus revealed complete absence of stem borer infestation in all the transgenic entries of ‘Hishell’ and ‘900M Gold’ with less than one leaf injury score (LIS), while in other conventional entries, stem borer infestation was observed and LIS was more than one. Hlicovepa armigera infestation was also not observed on transgenic hybrids whereas in the remaining non-transgenic maize treatment, significant attack of Helicoverpa armigera was observed in the range of 37 to 56%. These transgenic entries were not found resistant to aphids and grass hoppers. Beneficial insects were observed to visit transgenic Bt maize and conventional maize entries with no significant difference.

Agronomic performance of transgenic and isogenic corn hybrids in the state of Mato Grosso do Sul1

ABSTRACT Theoretically, isogenic hybrids should not present any differences in their genetic makeup due to recombinant DNA technology. However, these materials may behave differently in relation to adaptability and stability. This study aimed at evaluating the agronomic performance of transgenic corn and their isogenic hybrids in two locations in Mato Grosso do Sul, Brazil. Three isogenic simple hybrids and two transgenic varieties of each hybrid were evaluated. The first and second corn crop of 2013/14 were simultaneously analyzed in Dourados and Caarapó, two locations in the State of Mato Grosso do Sul, totaling four environments x season conditions that were evaluated.. A randomized complete block design with three replicates per location was used, and the treatments were arranged in a 3 × 3 factorial design (three genetically modified and three conventional hybrids), with three replicates per site. Characteristics such as plant height, ear height, male flowers, female flowers, and grain yield were evaluated. The parameters adaptability and stability were estimated using the Eberhart and Russell model. In all environments, transgenic hybrids showed higher average grain yield. Considering the environments, AG 7000, a conventional isogenic hybrid, obtained the highest average grain yield. The most stable and productive hybrids were DKB 390 VT PRO and AG 7000 YG, both transgenic. All assessed hybrids exhibited greater average plant height, ear height, and grain yield in Caarapó during the second corn crop, demonstrating that the environment at that site and time was more favorable.

SSR analysis on stress effect of transgenic hybrid poplar 741 on Clostera anachoreta (Fabricius) (Lepidoptera: Notodontidae)

The genetic differentiation of the experimental population of Clostera anachoreta fed on different resistant transgenic 741 poplar leaves was analyzed by SSR molecular marker technique to investigate stress effect of transgenic poplar Bt gene as food on target insect. The experimental population of C. anachoreta fed on transgenic 741 poplar high resistant strains 'Pb29', medium resis-tant strains 'Pb17' and non-transgenic poplar (CK), and the screened ten pairs of SSR primers were used. The results showed that 76 alleles were observed in ten pairs of primers. The average allele was 7.6, the average effective number of alleles was 2.2, the average observed heterozygosity was 0.5167, the average expected heterozygosity was 0.5167, and the average percentage of polymorphic loci was 96.7%. The genetic diversity level of C. anachoreta experimental population fed on transgenic poplar 741 was significantly higher than that fed on non-transgenic populations, and C. anachoreta fed on high resistance had the lowest genetic similarity with CK samples, which showed an increasing trend of the genetic diversity of the experimental population fed on transgenic Bt poplar. It was thus clear that transgenic hybrid poplar 741 had stress effects on genetic differentiation of C. anachoreta experimental population by SSR.

Levels of DNA methylation and transcript accumulation in leaves of transgenic maize varieties.

BackgroundPrior to their release in the environment, transgenic crops are examined for their health and environmental safety. In addition, transgene expression needs to be consistent in order to express the introduced trait (e.g. insecticidal and/or herbicide tolerance). Moreover, data on expression levels for GM events are usually required for approval, but these are rarely disclosed or they are considered insufficient. On the other hand, biosafety regulators do not consider epigenetic regulation (e.g. DNA methylation, ncRNAs and histone modifications), which are broadly known to affect gene expression, within their risk assessment analyses. Here we report the results of a DNA methylation (bisulfite sequencing) and transgene transcript accumulation (RT-qPCR) analysis of four Bt-expressing single transgenic maize hybrids, under different genetic backgrounds, and a stacked transgenic hybrid expressing both insecticidal and herbicide tolerance traits.ResultsOur results showed differences in cytosine methylation levels in the FMV promoter and cry2Ab2 transgene of the four Bt-expressing hybrid varieties. The comparison between single and stacked hybrids under the same genetic background showed differences in the 35S promoter sequence. The results of transgene transcript accumulation levels showed differences in both cry1A.105 and cry2Ab2 transgenes among the four Bt-expressing hybrid varieties. The comparison between single and stacked hybrids showed difference for the cry2Ab2 transgene only.ConclusionsOverall, our results show differences in DNA methylation patterns in all varieties, as well as in transgene transcript accumulation levels. Although the detection of changes in DNA methylation and transgenic accumulation levels does not present a safety issue per se, it demonstrates the need for additional studies that focus on detecting possible safety implications of such changes.Electronic supplementary materialThe online version of this article (doi:10.1186/s12302-016-0097-2) contains supplementary material, which is available to authorized users.

0639 WS Immunodetection of the Cry toxin in leaves of transgenic maize hybrids

0639 WS Immunodetection of the Cry toxin in leaves of transgenic maize hybrids

Communities of endophytic microorganisms in different developmental stages from a local variety as well as transgenic and conventional isogenic hybrids of maize.

The diversity of endophytic microorganisms may change due to the genotype of the host plant and its phenological stage. In this study we evaluated the effect of phenological stage, transgenes and genetic composition of maize on endophytic bacterial and fungal communities. The maize populations were composed of a local variety named Rosado (RS) and three isogenic hybrids. One isogenic hybrid was not genetically modified (NGM). Another hybrid (Hx) contained the transgenes cry1F and pat (T1507 event), which provide resistance to insects of the order Lepidoptera and tolerance to the glufosinate-ammonium herbicide, respectively. The third hybrid (Hxrr) contained the transgene cp4 epsps (NK603 event) combined with the transgenes cry1F and pat (T1507 event), which allow tolerance to the Roundup Ready herbicide, besides the characteristics of Hx. Evaluation of the foliar tissue was done through PCR-DGGE analysis, with specific primers for bacteria and fungi within four phenological stages of maize. The endophytic bacteria were only clustered by phenological stages; the structure of the fungal community was clustered by maize genotypes in each phenological stage. The fungal community from the local variety RS was different from the three hybrids (NGM, Hx and Hxrr) within the four evaluated stages. In the reproductive stage, the fungal community from the two transgenic hybrids (Hx and Hxrr) were separated, and the Hxrr was different from NGM, in the two field experiments. This research study showed that the genetic composition of the maize populations, especially the presence of transgenes, is the determining factor for the changes detected in the endophytic fungal community of maize leaves.

Integrated Fertilizer Prescriptions for Transgenic Cotton Hybrids under Rainfed Situation through Inductive cum Targeted Yield Model on Vertisol

ABSTRACTThe fertilizer prescription equations were developed for rainfed Bt cotton on Pielamedu series (Typic haplusterts) by adopting the inductive cum targeted yield model, and fertilizer requirements were quantified for Bt cotton based on soil test and yield target. The validity of fertilizer prescription equations was done by conducting test verification trials at four locations in Perambalur district of Tamil Nadu developed for rainfed Bt cotton. The fertilizer prescription equations were used to calculate the fertilizer doses for STCR treatments. The percent achievement of the targeted yield was within ±10% variation in all the locations for both the yield targets under nitrogen, phosphorus, and potassium (NPK) alone and IPNS, proving the validity of the equations. STCR-IPNS based fertilizer recommendations for a yield target of 32 q/ha has recorded the highest yield, nutrient response ratio, and percent yield achievement in all the locations.

English

The objective of this study was to evaluate resistance mechanisms in 12 corn genotypes (transgenic hybrids: 30A91 PW, 20A78 HX, Impacto VIP 3, 20A55 HX, NS90 PRO 2, Maximus VIP 3, BX 1293YG, RB 9004 PRO, Feroz VIP 3, LG 6036 PRO; conventional: AG 1051 and variety: AL Bandeitante). Attractiveness, aversion to feeding, and antibiosis were evaluated via free and no choice tests in the laboratory. Attractiveness was evaluated at 1, 5, 10, 15 and 30 min and 1, 2, 6, 12 and 24 h by counting the number of larvae that fed on each genotype. The preference for feeding was determined by quantifying the leaf area of each genotype consumed. Antibiosis was determined by assessing biological parameters of the fed caterpillars in relation to each genotype. The biological parameters evaluated were (a) Larval stage: the viability of the larval stage and weight of larvae at ten days; and (b) pupal stage: the viability and weight of pupae at 24 h of age. After emergence, the moths were fed and evaluated to assess the longevity of the adults and the total life cycle. The transgenic genotypes NS90 PRO2, Maximus VIP 3, Feroz VIP 3 and Maximus VIP 3 elicited an aversion and/or an antibiosis reaction from fall armyworm (FAW). The transgenic genotypes HX 20A55, 30A91 PW, LG 6036 PRO, 20A78 HX and BR 9004 PRO showed a moderate resistance to FAW. The conventional genotypes AG 1051 and AL Bandeirante were highly susceptible to FAW and the transgenic genotype BX 1293 YG was susceptible to FAW. Key words: Integrated pest management, transgenic crop, plant resistance to insects, Zea mays.

Expression of Bacillus thuringiensis cytolytic toxin (Cyt2Ca1) in citrus roots to control Diaprepes abbreviatus larvae

Diaprepes abbreviatus (L.) is an important pest of citrus in the USA. Currently, no effective management strategies of D. abbreviatus exist in citriculture, and new methods of control are desperately sought. To protect citrus against D. abbreviatus a transgenic citrus rootstock expressing Bacillus thuringiensis Cyt2Ca1, an insect toxin protein, was developed using Agrobacterium-mediated transformation of ‘Carrizo’ citrange [Citrus sinensis (L) Osbeck Poncirus trifoliate (L) Raf]. The transgenic citrus root stock expressed the cytolytic toxin Cyt2Ca1 constitutively under the control of a 35S promoter in the transgenic Carrizo citrange trifoliate hybrid including the roots that are the food source of larval D. abbreviatus. The engineered citrus was screened by Western blot and RT-qPCR analyses for cyt2Ca1 and positive citrus identified. Citrus trees expressing different levels of cyt2Ca1 transcripts were identified (Groups A–C). High expression of the toxin in the leaves (109 transcripts/ng RNA), however, retarded plant growth. The transgenic plants were grown in pots and the roots exposed to 3week old D. abbreviatus larvae using no-choice plant bioassays. Three cyt2Ca1 transgenic plants were identified that sustained less root damage belonging to Group B and C. One plant caused death to 43% of the larvae that fed on its roots expressed 8×106cyt2Ca1 transcripts/ng RNA. These results show, for the first time, that Cyt2Ca1 expressed in moderate amounts by the roots of citrus does not retard citrus growth and can protect it from larval D. abbreviatus.

Neutralization of Bacterial YoeBSpn Toxicity and Enhanced Plant Growth in Arabidopsis thaliana via Co-Expression of the Toxin-Antitoxin Genes.

Bacterial toxin-antitoxin (TA) systems have various cellular functions, including as part of the general stress response. The genome of the Gram-positive human pathogen Streptococcus pneumoniae harbors several putative TA systems, including yefM-yoeBSpn, which is one of four systems that had been demonstrated to be biologically functional. Overexpression of the yoeBSpn toxin gene resulted in cell stasis and eventually cell death in its native host, as well as in Escherichia coli. Our previous work showed that induced expression of a yoeBSpn toxin-Green Fluorescent Protein (GFP) fusion gene apparently triggered apoptosis and was lethal in the model plant, Arabidopsis thaliana. In this study, we investigated the effects of co-expression of the yefMSpn antitoxin and yoeBSpn toxin-GFP fusion in transgenic A. thaliana. When co-expressed in Arabidopsis, the YefMSpn antitoxin was found to neutralize the toxicity of YoeBSpn-GFP. Interestingly, the inducible expression of both yefMSpn antitoxin and yoeBSpn toxin-GFP fusion in transgenic hybrid Arabidopsis resulted in larger rosette leaves and taller plants with a higher number of inflorescence stems and increased silique production. To our knowledge, this is the first demonstration of a prokaryotic antitoxin neutralizing its cognate toxin in plant cells.

Ambient insect pressure and recipient genotypes determine fecundity of transgenic crop-weed rice hybrid progeny: Implications for environmental biosafety assessment.

Transgene introgression into crop weedy/wild relatives can provide natural selective advantages, probably causing undesirable environmental impact. The advantages are likely associated with factors such as transgenes, selective pressure, and genetic background of transgene recipients. To explore the role of the environment and background of transgene recipients in affecting the advantages, we estimated the fitness of crop‐weed hybrid lineages derived from crosses between marker‐free insect‐resistant transgenic (Bt/CpTI) rice with five weedy rice populations under varied insect pressure. Multiway anova indicated the significant effect of both transgenes and weedy rice genotypes on the performance of crop‐weed hybrid lineages in the high‐insect environment. Increased fecundity was detected in most transgene‐present F1 and F2 hybrid lineages under high‐insect pressure, but varied among crop‐weed hybrid lineages with different weedy rice parents. Increased fecundity of transgenic crop‐weed hybrid lineages was associated with the environmental insect pressure and genotypes of their weedy rice parents. The findings suggest that the fitness effects of an insect‐resistant transgene introgressed into weedy populations are not uniform across different environments and genotypes of the recipient plants that have acquired the transgene. Therefore, these factors should be considered when assessing the environmental impact of transgene flow to weedy or wild rice relatives.

Potential of genetically engineered hybrid poplar for pyrolytic production of bio-based phenolic compounds

Wild-type and two genetically engineered hybrid poplar lines were pyrolyzed in a micro-pyrolysis (Py-GC/MS) and a bench scale setup for fast and intermediate pyrolysis studies. Principal component analysis showed that the pyrolysis vapors obtained by micro-pyrolysis from wood of caffeic acid O-methyltransferase (COMT) and caffeoyl-CoA O-methyltransferase (CCoAOMT) down-regulated poplar trees differed significantly from the pyrolysis vapors obtained from non-transgenic control trees. Both fast micro-pyrolysis and intermediate pyrolysis of transgenic hybrid poplars showed that down-regulation of COMT can enhance the relative yield of guaiacyl lignin-derived products, while the relative yield of syringyl lignin-derived products was up to a factor 3 lower. This study indicates that lignin engineering via genetic modifications of genes involved in the phenylpropanoid and monolignol biosynthetic pathways can help to steer the pyrolytic production of guaiacyl and syringyl lignin-derived phenolic compounds such as guaiacol, 4-methylguaiacol, 4-ethylguaiacol, 4-vinylguaiacol, syringol, 4-vinylsyringol, and syringaldehyde present in the bio-oil.

Hybridization of downregulated-COMT transgenic switchgrass lines with field-selected switchgrass for improved biomass traits

Transgenic switchgrass (Panicum virgatum L.) has been produced for improved cell walls for biofuels. For instance, downregulated caffeic acid 3-O-methyltransferase (COMT) switchgrass produced significantly more biomass and biofuel than the non-transgenic progenitor line. In the present study we sought to further improve biomass characteristics by crossing the downregulated COMT T1 lines with high-yielding switchgrass accessions in two genetic backgrounds (‘Alamo’ and ‘Kanlow’). Crosses and T2 progeny analyses were made under greenhouse conditions to assess maternal effects, plant morphology and yield, and cell wall traits. Female parent type influenced morphology, but had no effect on cell wall traits. T2 hybrids produced with T1 COMT-downregulated switchgrass as the female parent were taller, produced more tillers, and produced 63 % more biomass compared with those produced using the field selected accession as the female parent. Transgene status (presence or absence of transgene) influenced both growth and cell wall traits. T2 transgenic hybrids were 7 % shorter 80 days after sowing and produced 43 % less biomass than non-transgenic null-segregant hybrids. Cell wall-related differences included lower lignin content, reduced syringyl-to-guaiacyl (S/G) lignin monomer ratio, and a 12 % increase in total sugar release in the T2 transgenic hybrids compared to non-transgenic null segregants. This is the first study to evaluate the feasibility of transferring the low-recalcitrance traits associated with a transgenic switchgrass line into high-yielding field varieties in an attempt to improve growth-related traits. Our results provide insights into the possible improvement of switchgrass productivity via biotechnology paired with plant breeding.

FUM Gene Expression Profile and Fumonisin Production by Fusarium verticillioides Inoculated in Bt and Non-Bt Maize.

This study aimed to determine the levels of fumonisins produced by Fusarium verticillioides and FUM gene expression on Bt (Bacillus thuringiensis) and non-Bt maize, post harvest, during different periods of incubation. Transgenic hybrids 30F35 YG, 2B710 Hx and their isogenic (30F35 and 2B710) were collected from the field and a subset of 30 samples selected for the experiments. Maize samples were sterilized by gamma radiation at a dose of 20 kGy. Samples were then inoculated with F. verticillioides and analyzed under controlled conditions of temperature and relative humidity for fumonisin B1 and B2 (FB1 and FB2) production and FUM1, FUM3, FUM6, FUM7, FUM8, FUM13, FUM14, FUM15, and FUM19 expression. 2B710 Hx and 30F35 YG kernel samples were virtually intact when compared to the non-Bt hybrids that came from the field. Statistical analysis showed that FB1 production was significantly lower in 30F35 YG and 2B710 Hx than in the 30F35 and 2B710 hybrids (P < 0.05). However, there was no statistical difference for FB2 production (P > 0.05). The kernel injuries observed in the non-Bt samples have possibly facilitated F. verticillioides penetration and promoted FB1 production under controlled conditions. FUM genes were expressed by F. verticillioides in all of the samples. However, there was indication of lower expression of a few FUM genes in the Bt hybrids; and a weak association between FB1 production and the relative expression of some of the FUM genes were observed in the 30F35 YG hybrid.