Bt Gene Research Articles

Generation of selectable marker-free transgenic rice resistant to chewing insects using two co-transformation systems

To produce selectable marker-free (SMF) transgenic rice resistant to chewing insects, the Bacillus thuringiensis cryIA(c) gene ( Bt) was introduced into two elite japonica rice varieties by using two Agrobacterium-mediated co-transformation systems. One system is with a single mini-twin T-DNA binary vector in one Agrobacterium strain, which consists of two separate T-DNA regions, one carrying the Bt while the other contains the selectable marker gene, hygromycin resistant gene ( HPT). The other system uses two separate binary vectors in two separate Agrobacterium cultures, containing the Bt or HPT gene on individual plasmids. A lot of independent transgenic rice lines harboring both Bt and selectable marker genes were obtained. The results showed that the co-transformation frequency of the Bt gene and HPT gene was much higher by using the mini-twin T-DNA vector system (29.87%) than that by the two separate binary vector systems (4.52%). However, the frequency of the SMF transgenic rice plants obtained from the offspring of co-transgenic plants (21.74%) was lower for the mini-twin T-DNA vector system than that for the latter (50–60%). The data of ELISA implied that the expressed Bt proteins were quantitated as 0.025–0.103% of total leaf soluble proteins in the transgenic plant. Therefore, several elite transgenic rice lines, free of the selectable marker gene, were chosen. The results from both in vitro and in vivo insect bioassays indicated that the SMF transgenic rice was shown to be highly resistant to the striped stem borer and rice leaf folder. Moreover, in a natural field condition without any insecticide applied, all the transgenic rice plants were found to be not injured by the rice leaf folder, whereas the wild types were impaired seriously.

Identification of resistance to corn-borer of transgenic maize inbred lines and hybrids with GFM Cry1A gene

AbstractSix transgenic maize lines with the fully modified gene GFM Cry1A were obtained and polymerase chain reaction (PCR) analysis of their T5 generation plants indicated that foreign genes could be stably inherited. Three hybrids (Simi25Bt, Tongdan24Bt and Jidan209Bt) were created using the transgenic lines and combined with regular maize inbred lines. The results of resistance identification of the transgenic inbred lines and hybrids showed that the difference of resistance among transgenic lines was very significant; there also existed a difference among individuals of the same line and among the three hybrids. Compared with the control inbred line (CK), the average of four transgenic inbred lines in the number of tunnels per stalk, larval tunnel length per plant and length of surviving larvae were decreased by 70%, 80% and 70%, respectively. Enzyme-linked immunosorbent assay (ELISA) testing showed that the average of Bt Cry1A protein expression level for six transgenic inbred lines was 0.16% total protein and the highest expression level was 0.19%. At maturity, compared with the control variety (CK), the larval tunnel length per plant of the three hybrids (Simi25Bt, Tongdan24Bt and Jidan209Bt) was decreased by 39.97%, 36.20% and 53.83%, respectively, which was a decrease of 43.36% on average. The investigation of agronomic characters showed that there was no significant difference between the improved hybrids and the control in plant height, ear length, row number per ear, kernel number per row and 100-kernels weight. It is thought that the GFM Cry1A gene can be applied to improve resistance to corn-borer, and maize inbred lines with the Bt gene can be directly applied to conventional maize breeding.

Several methods to detect the inheritance and resistance to the diamondback moth in transgenic Chinese cabbage.

A positive selection system was developed for Agrobacterium-mediated trasnsformation of using hypocotyl segments of aseptic seedlings of chinese cabbage as explants, regenerated plants with kanamycin resistance were obtained. The transformed plants with CryIA(c) (Bt) gene were confirmed by Southern blotting analysis, indicating the integration of the transgene into the cabbage genome. Majority of the transgenic plants had only a single copy of the inserted CryIA(c) gene. Leaf section bioassays showed that resistance against larvae of diamondback moth in CryIA(c) transgenic cabbage was significantly enhanced. The inheritance patterns of the transgene in T1 offspring of transgenic cabbage were investigated using PCR analysis and kanamycin resistance test on young seedling leaves.

Cry2Ab Tolerance Response of <I>Helicoverpa armigera</I> (Lepidoptera: Noctuidae) Populations From Cry1Ac Cotton Planting Region

Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) is an important lepidopteran pest of cotton, Gossypium hirsutum L., in Asia. Transgenic cotton expressing the cry1Ac gene from Bacillus thuringiensis (Bt) has been widely planted for control of this pest. For managing the potential risk from resistance evolution in this pest, a new transgenic Bt cotton containing cry1Ac and cry2Ab genes (gene pyramided strategy) was designed as a successor of cry1Ac cotton. This article reports on levels of Cry2Ab tolerance in H. armigera populations from CrylAc cotton planting region in China based on bioassays of F1 and F2 offspring of isofemale lines. In total, 572 isofemale families of H. armigera from Xiajin County of Shandong Province (an intensive Bt cotton-planting area) and 124 families from Anci County of Hebei Province [a multiple-crop system, including corn (Zea mays L.), soybean (Gycine max (L.) Merr., peanut (Arachis spp.), and Bt cotton] were screened with both Cry1Ac- and Cry2Ab-containing diets in 2008. The bioassays results indicated that relative average development rates (RADR) of F1 full-sib families from field-collected female moths on Cry1Ac- and Cry2Ab-containing diet were positively correlated. The same correlation was found in the F2 generation, indicating cross-tolerance between Cry1Ac and Cry2Ab in field populations of H. armigera in Yellow River cotton-farming region of China. This cross-tolerance must be considered in evaluating the utility of pyramiding Bt genes in cotton for delaying evolution of resistance.

Effects of Bt transgenic crops on soil ecosystems: a review of a ten-year research in China

Bacillus thuringiensis (Bt) transgenic cotton is the unique Bt transgenic crop planted on a large scale in China, and its commercialized varieties and hectareage had increased rapidly in China during the past decade (1997–2006) with broad geographic distribution for the economic, environmental, and health benefits. In 2004, the planting area of Bt transgenic cotton in China ranked first worldwide with up to 370 × 106 hm2. In addition, Bt transgenic rice varieties in field tests have been close to approval for commercialization. However, ecological risks, a complex issue of Bt transgenic crops on soil ecosystem is urgently faced in China due to more than 60 varieties transferred single or bivalent Bt genes grown under diverse geographic regions. Two main pathways, biomass incorporation and root exudates, are involved in the effects of Bt transgenic crops on soil ecosystems. In this paper, the research results in recent years in China involved in the effects of Bt transgenic crops (Bt transgenic cottons and rice) on soil ecosystems were summarized with special attentions paid to the release and persistence of Bt toxins, and the toxicology to microorganisms, as well as the change of soil biochemical properties in soils where Bt transgenic crops were planted or incubated with their biomass. In addition, the complexity and current research defaults of ecological risk evaluation of Bt transgenic crops in China were highlighted.

Effect of introducing Bt gene in cotton on biochemical content of vascular bundle sap of cotton leaf and reproduction of <I>Bemisia tabaci</I> (Gennadius)

Effect of introducing Bt gene in cotton on biochemical content of vascular bundle sap of cotton leaf and reproduction of <I>Bemisia tabaci</I> (Gennadius)

Mineral element distribution in organs of dual-toxin transgenic (Bt+CpTI) cotton seedling

Being a new cultivar, the physiology of transgenic cotton, especially dual-toxin transgenic (Bt+CpTI) cotton, is not yet completely understood. Twelve elements in three organs of dual-toxin transgenic cotton seedlings were analyzed by ICP-MS. The distributions of the 12 elements were substantially different from those of non-transgenic cotton. In particular, the contents of B, Mg, P, K and Ca were the highest in leaves, while those of Si, Fe, Rb and Cu were the highest in roots; other elements had similar contents in the two organs, which were higher than those in the stem. Compared with non-transgenic cotton, the 12 elements could be classified into four groups according to their contents and distributions in the three organs: (a) P, K and Cu: their contents in transgenic cotton were remarkably lower, especially contents of P and K in leaves that were one times lower than those in leaves of non-transgenic cotton; (b) B, Mg and Mo: their contents in leaves and roots of transgenic cotton were higher, but lower in stems, compared with non-transgenic cotton; (c) Si, Mn, Fe, Rb and Zn: compared with non-transgenic cotton, these were lower in leaves and stems, but higher in roots of transgenic cotton; and (d) Ca: compared with non-transgenic cotton, its content was higher in all three organs of the transgenic counterpart. The decrease in soluble proteins and the expression of Bt and CpTI genes could be responsible for these changes. Further studies are needed to verify this hypothesis.

Transgenic resistance of Bulgarian potato cultivars to the Colorado potato beetle based on Bt technology

Colorado potato beetle, Leptinotarsa decemlineata Say, is the most destructive insect pest of potatoes. When the population of beetles is high, plants can be completely defoliated and commercial potato production is nearly impossible without control of the beetle. The beetles have shown a tremendous ability to develop resistance against insecticides. Previously, a biotechnology approach to control Colarado potato beetle based on the use of the synthetic Bt gene was developed. In this article, a transformation procedure for three commercial Bulgarian potato cultivars was developed and potentially commercial transgenic lines have been selected based on field resistance to Colorado potato beetles and yield. Plants were transformed with the Bacillus thuringiensis (Bt) cry3A gene using Agrobacterium-mediated transformation. 110 plants from the three cultivars were regenerated and tested by double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA). The Cry3A protein accumulation varied across the transgenic lines, rating from very low to 71.5 μg/g fresh weight. 21 transgenic lines expressing the Cry3A protein at levels above 10 μg/g fresh weight were tested in two successive years in filed conditions at two different locations of the country. All transgenic lines compared with the controls, nontransgenic potatoes from the respective cultivar, were consistently protected from foliar damages from all developmental stages of the beetle. The comparison of all properties of the tested transgenic lines, including variety phenotypes and tuber yield, allowed the selection of the most promising 2–3 lines per cultivar. Selected lines produced tuber amounts 80–100% higher compared with the control, non-transgenic plants. Those lines were grown for mass propagation during the third year of field experiments. The presence of the transgene in these lines was confirmed with the use of primers specific to the transgene by polymerase chain reaction (PCR). Additionally, the results from the insect bioassay showed that these lines were highly resistant to insect feeding, leading to 100% of mortality of larval populations. In summary, we generated potentially commercial potato lines highly resistant to Colorado potato beetle using Bt technology that may have a profound impact on development of sustainable agriculture in Bulgaria. This is one of the several agriculture biotechnology products entirely developed and tested in Bulgaria.

Occurrence of Bacillus thuringiensis harboring insecticidal cry1 genes in a corn field in Northern Italy

Bacillus thuringiensis (Bt) is a ubiquitous bacteria widely used as a biopesticide to control a number of important insect pests. Since the mid-1990s, genetically modified (GM) plants expressing Bt genes have been used as an effective tool to control a wide range of insect pests. In recent years, a wide number of articles addressing the environmental impact of genetically modified plants have been published. However, only a few have addressed the occurrence and distribution of the indigenous Bt population in agricultural systems. Here, culturing and molecular methods were used to study the occurrence of Bt harboring insecticidal cry1 genes in a corn field. Samples of corn leaves and soil were collected in July and August 2007 from a 10 ha corn field in Northern Italy. The results showed that the highest Bt density was in leaves located near the soil surface. The incidence of Bt isolates harboring antilepidopteran cry1 genes was 42% of the total tested isolates. Approximately 20% of the Bt isolates harbored the cry1Ab and cry1Ac genes. Similarly to Bt density, the highest abundance of isolates harboring cry1 genes was found in leaves collected near the plant collar. Less than 9% of the Bt isolated from soil harbored cry1 genes. Density of Bt was reduced by application of the insecticide chlorpiryfos. This effect appeared to be due to simply washing off effects of the insecticide treatment to the corn phyllosphere. This study showed that Bt was fairly abundant in the corn agroecosystem and that the high incidence of isolates harboring antilepidopterean cry1 genes could have a role in preserving the sustainability of the agroecosystem.

No Evidence ThatBacillus thuringiensisGenes and Their Products Influence the Susceptibility of Corn Residue to Decomposition

The possibility thatBacillus thuringiensis(Bt) corn (Zea maysL.) residues resist decomposition compared to non‐Bt residues would present direct (soil carbon turnover times) and indirect (changes in tillage practices) effects on carbon budgets in agricultural systems. We evaluated the relative decomposition of residue from two pairs of Bt and non‐Bt corn hybrids from different seed manufacturers buried in the root zone of adjacent Bt and non‐Bt corn plots over a period of 384 d. We found no persistent differences in residue decomposition among the different hybrids regardless of the seed manufacturer or the presence of the Bt genes (bothcry1Abandcry3Bb1genes present in each Bt hybrid) in the residue. No significant differences in residue compositional properties or flexural strength of intact stalk sections were observed among the four hybrids. Both Bt and non‐Bt residues buried in the root zone of a Bt corn hybrid decomposed faster than those buried in the root zone of the corresponding, near‐isogenic non‐Bt hybrid. A subsequent replicated laboratory study showed no difference between the decomposition of cellulose filter paper buried in the root zone of growing and senescing Bt and non‐Bt hybrids. We conclude that (i) the presence of Bt genes and their products in chopped residue does not affect its decomposability; (ii) the presence of Bt genes does not affect the mechanical strength of the stalks; and, (iii) Bt products released from growing or senescing Bt corn plants do not adversely affect decomposition activities in the surrounding soil.

Consumer Acceptance of Genetically Modified (GM)—Cowpeas in Sub-Sahara Africa

ABSTRACT Cowpea (known as black-eyed peas in America) is the most important indigenous African grain legume for both home use and as a cash crop. Because of its tolerance to drought, it is especially important for the Sahel. Genetic transformation of cowpea with Bacillus thuringiensis (Bt) genes to control pod-boring insects has many advantages, but little is known of the potential consumer response. This article analyzes and reports the results of a survey of 200 consumers in northern Nigeria concerning consumer awareness of and acceptance of biotechnology. Ninety percent of the respondents were aware of genetically modified (GM) products. Those respondents who were most concerned about the ethics of genetic transformation were likely to disapprove of such products, while those individuals who identified international radio as an information source were more likely to approve of GM technology.

Cellular automata-based modeling and risk assessment of genetically modified crops with Bacillus thuringiensis (bt) genes

Cellular automata-based modeling and risk assessment of genetically modified crops with Bacillus thuringiensis (bt) genes

Transformation and inheritance of Bt genes inGossypium hirsutum

Transgenic plants offer many unique opportunities for managing pest populations. However, the inheritance, integration, and expression of multiple transgenes are prerequisite for maintaining sustainable resistance against insects in crops. We took a gene-pyramiding approach to produce Bt cotton expressing two Bt genes,cry1Ac andcry2A. Using sonication-assistedAgrobacterium-mediated transformation (SAAT), we achieved an efficiency of 6.26%. Putative transgenic plants were confirmed via PCR, Southern hybridization, and western-blotting. Those showing mortality of 75 to 100% for the second instar ofHeliothis armigera (compared with 0% for the control) were considered Bt-positive. Transgenes were segregated according to a 3:1 Mendelian inheritance pattern in the T1 generation forHeliothis resistance. In our insect bioassay, the control plants showed >95% leaf damage, and insects reached the 4th instar stage of larval growth. In contrast, leaf damage on transgenic plants was limited to only a few bites, and insect mortality was 75 to 100%. ELISA confirmed transgene expression, and Bt protein was detected in leaf tissue. This performance was consistent with that of the parent transgenics. PCR and Southern blots verified integration of thecry1Ac andcry2A genes into the progeny. Therefore, this strategy provides a pathway toward cotton improvement and the development of durable resistance against insect damage.

Transgenic tobacco plants expressing synthetic Cry1Ac and Cry1Ie genes are more toxic to cotton bollworm than those containing one gene

Transgenic tobacco plants carrying Cry1Ac, Cry1Ie or both genes were obtained. In the leaves of transgenic plants carrying both genes, the contents of Cry1Ac and Cry1Ie proteins were 0.173% and 0.131% of the total proteins, respectively. Cry1Ac protein content was 0.182% and Cry1Ie protein content was 0.124% of the total proteins in the leaves of transgenic plants containing only one Bt gene. Fresh leaves of transgenic tobacco and wild-type plants were used for the insect bioassay against wild-type and Cry1Ac-resistant cotton bollworm (Helicoverpa armigera). The bioassay results showed that transgenic plants carrying both genes were significantly more toxic to wild-type and Cry1Ac-resistant cotton bollworm than those carrying Cry1Ac or Cry1Ie alone. This study indicates that the higher toxicity of transgenic tobacco plants carrying both genes is caused by the cooperative function of both Bt proteins, thus providing a potential way to delay the development of insect resistance to transgenic crops.

Effect of Transgenic <I>Bacillus thuringiensis</I> Rice Lines on Mortality and Feeding Behavior of Rice Stem Borers (Lepidoptera: Crambidae)

Ten transgenic Bacillus thuringiensis Bt rice, Oryza sativa L., lines with different Bt genes (two Cry1Ac lines, three Cry2A lines, and five Cry9C lines) derived from the same variety Minghui 63 were evaluated in both the laboratory and the field. Bioassays were conducted by using the first instars of two main rice lepidopteran insect species: yellow stem borer, Scirpophaga incertulas (Walker) and Asiatic rice borer, Chilo suppressalis (Walker). All transgenic lines exhibited high toxicity to these two rice borers. Field evaluation results also showed that all transgenic lines were highly insect resistant with both natural infestation and manual infestation of the neonate larvae of S. incertulas compared with the nontransformed Minghui63. Bt protein concentrations in leaves of 10 transgenic rice lines were estimated by the sandwich enzyme-linked immunosorbent assay. The cry9C gene had the highest expression level, next was cry2A gene, and the cry1Ac gene expressed at the lowest level. The feeding behavior of 7-d-old Asiatic rice borer to three classes of Bt transgenic rice lines also was detected by using rice culm cuttings. The results showed that 7-d-old larvae of Asiatic rice borer have the capacity to distinguish Bt and non-Bt culm cuttings and preferentially fed on non-Bt cuttings. When only Bt culm cuttings with three classes of different Bt proteins (CrylAc, Cry2A, and Cry9C) were fed, significant distribution difference of 7-d-old Asiatic rice borer in culm cuttings of different Bt proteins also was found. In the current study, we evaluate different Bt genes in the same rice variety in both the laboratory and the field, and also tested feeding behavior of rice insect to these Bt rice. These data are valuable for the further development of two-toxin Bt rice and establishment of appropriate insect resistance management in the future.

Effect of Transgenic Bacillus thuringiensis Rice Lines on Mortality and Feeding Behavior of Rice Stem Borers (Lepidoptera: Crambidae)

Ten transgenic Bacillus thuringiensis Bt rice, Oryza sativa L., lines with different Bt genes (two Cry1Ac lines, three Cry2A lines, and five Cry9C lines) derived from the same variety Minghui 63 were evaluated in both the laboratory and the field. Bioassays were conducted by using the first instars of two main rice lepidopteran insect species: yellow stem borer, Scirpophaga incertulas (Walker) and Asiatic rice borer, Chilo suppressalis (Walker). All transgenic lines exhibited high toxicity to these two rice borers. Field evaluation results also showed that all transgenic lines were highly insect resistant with both natural infestation and manual infestation of the neonate larvae of S. incertulas compared with the nontransformed Minghui63. Bt protein concentrations in leaves of 10 transgenic rice lines were estimated by the sandwich enzyme-linked immunosorbent assay. The cry9C gene had the highest expression level, next was cry2A gene, and the cry1Ac gene expressed at the lowest level. The feeding behavior of 7-d-old Asiatic rice borer to three classes of Bt transgenic rice lines also was detected by using rice culm cuttings. The results showed that 7-d-old larvae of Asiatic rice borer have the capacity to distinguish Bt and non-Bt culm cuttings and preferentially fed on non-Bt cuttings. When only Bt culm cuttings with three classes of different Bt proteins (Cry1Ac, Cry2A, and Cry9C) were fed, significant distribution difference of 7-d-old Asiatic rice borer in culm cuttings of different Bt proteins also was found. In the current study, we evaluate different Bt genes in the same rice variety in both the laboratory and the field, and also tested feeding behavior of rice insect to these Bt rice. These data are valuable for the further development of two-toxin Bt rice and establishment of appropriate insect resistance management in the future.

Production of Helicoverpa spp. (Lepidoptera, Noctuidae) from different refuge crops to accompany transgenic cotton plantings in eastern Australia

The adoption of transgenic (Bt) cotton varieties has markedly reduced feeding damage by noctuid pests, Helicoverpa armigera and H. punctigera, in Australian cotton crops. However, the potential for Bt resistance to evolve within these insect pests is a major concern. To reduce the likelihood of Bt resistance occurring, it is mandatory for growers of Bt cotton to also grow refuge crops which produce large numbers of Bt-susceptible moths. We present here findings from a series of field studies which sought to identify the relative Helicoverpa productivity of different refuge crop options. The abundance of Helicoverpa pupae (during the cotton season) was compared under crops of Ingard® (transgenic, single Bt gene) cotton, sprayed conventional cotton, and various unsprayed refuge crops (cotton, sorghum, maize, pigeon pea) from 1996 to 2003 in the major cotton-producing regions of northern New South Wales, plus St George and Dirranbandi in southern Queensland. Unsprayed, conventional cotton was used as the ‘control’ refuge in these studies. Productivity of adult Helicoverpa was estimated by surveying for pupae and discounting those that were parasitised and dead. Some supplementary experiments were also conducted to evaluate specific issues, in particular the value of additional refuge crops sown late in 2000–01 (due to accidental over-planting of Ingard® cotton that year) and the potential productivity of mixed plantings of various refuge crops which individually flowered at different times throughout the cotton season. Unsprayed refuges generally produced many more pupae than sprayed, conventional cotton and Ingard® cotton. Overall, pigeon pea, which has the advantage of prolonged flowering and thus on-going attractiveness to Helicoverpa, was the most reliable and productive refuge option, producing about twice as many pupae as unsprayed cotton. The seed mix refuge is perhaps a viable alternative option, although logistically more difficult for growers to adopt. Laboratory rearing of live pupae until moth emergence indicated that parasitism can be a substantial source of mortality in some refuge crops, especially sorghum. The most common parasitoid of Helicoverpa pupae was the ichneumonid wasp, Heteropelma scaposum. Such parasitism can greatly reduce productivity of adult Helicoverpa from refuges and needs to be taken into account when assessing relative refuge ‘value’, although such refuges will obviously contribute to the abundance of these beneficial species across the landscape.

Effects of defoliating insect resistance QTLs and a cry1Ac transgene in soybean near-isogenic lines

The crystal proteins coded by transgenes from Bacillus thuringiensis (Bt) have shown considerable value in providing effective insect resistance in a number of crop species, including soybean, Glycine max (L.) Merr. Additional sources of soybean insect resistance would be desirable to manage the development of tolerance/resistance to crystal proteins by defoliating insects and to sustain the deployment of Bt crops. The objective of this study was to evaluate the effects and interactions of three insect resistance quantitative trait loci (QTLs; QTL-M, QTL-H, and QTL-G) originating from Japanese soybean PI 229358 and a cry1Ac gene in a "Benning" genetic background. A set of 16 BC(6)F(2)-derived near isogenic lines (NILs) was developed using marker-assisted backcrosses and evaluated for resistance to soybean looper [SBL, Pseudoplusia includens (Walker)] and corn earworm [CEW, Helicoverpa zea (Boddie)] in field cage, greenhouse, and detached leaf assays. Both Bt and QTL-M had significantly reduced defoliation by both SBL and CEW and reduced larval weight of CEW. The antibiosis QTL-G had a significant effect on reducing CEW larval weight and also a significant effect on reducing defoliation by SBL and CEW in some assays. The antixenosis QTL-H had no main effect, but it appeared to function through interaction with QTL-M and QTL-G. Adding QTL-H and QTL-G further enhanced the resistance of the Bt and QTL-M combination to CEW in the field cage assay. These results should help guide the development of strategies for effective management of insect pests and for sustainable deployment of Bt genes.

Effects of Exogenous Jasmonic Acid on Concentrations of Direct-Defense Chemicals and Expression of Related Genes in Bt ( Bacillus thuringiensis) Corn ( Zea mays)

Bt corn is one of the top three large-scale commercialized transgenic crops around the world. It is increasingly clear that the complementary durable approaches for pest control, which combine the endogenous defense of the crop with the introduced foreign genes, are promising alternative strategies for pest resistance management and the next generation of insect-resistant transgenic crops. In the present study, we tested the inducible effects of exogenous jasmonic acid (JA) on direct-defense chemical content, Bt protein concentration, and related gene expression in the leaves of Bt corn cultivar 34B24 and non-Bt cultivar 34B23 by chemical analysis, ELISA, and RT-PCR. The results show that the expression of LOX, PR-2α, MPI, and PR-1 genes in the treated leaf (the first leaf) was promoted by exogenous JA both in 34B24 and 34B23. As compared with the control, the concentration of DIMBOA in the treated leaf was significantly increased by 63 and 18% for 34B24 and 34B23, respectively. The total phenolic acid was also increased by 24 and 12% for both 34B24 and 34B23. The Bt protein content of 34B24 in the treated leaf was increased by 13% but decreased significantly by 27% in the second leaf. The induced response of 34B24 was in a systemic way and was much stronger than that of 34B23. Those findings indicated that there is a synergistic interaction between Bt gene and internally induced chemical defense system triggered by externally applied JA in Bt corn.

Transgenic tobacco plants containing Bt and GNA genes

A new plant expression vector (pBSbtCry1Ac-GNA) containing two insect resistant genes, a synthetic chimeric gene SbtCry1Ac encoding the insecticidal protein CrylAc and a gene GNA encoding snowdrop lectin (Galanthus nivalis agglutinin) was constructed. Transgenic tobacco plants containing these two genes were obtained through Agrobacterium-mediated transformation of tobacco leaf discs. Results from PCR detection and genomic DNA Southern blot analysis indicated that both SbtCrylAc gene and GNA gene were integrated into the genome of these plants. Results of Western blot analysis indicated that these two proteins were expressed in the analyzed plants. Bioassays of Myzus persicae and Helicoverpa assulta on detached leaves of transformed tobacco plants were carried out. The average aphid inhibition rate of these plants tested at 12 d post-infestation was 71.9 %. The average H. assulta mortality of these plants tested at 6 d post-infestation was up to 89.8 %. The kanamycin resistance of the T1 progeny of these transgenic plants was analyzed and a typical 3:1 segregation was observed.