To investigate the extent of exposure and routes of Cry1Ac1 protein through the food chain, we collected Bt cabbage leaves and arthropods that occurred in the field during two trials. Protein levels in the transgenic leaves were significantly higher during the early stages of plant growth, ranging from 209.1 to 553.6 ng g(-1) in spring and from 208.2 to 402.8 ng g(-1) in autumn. Enzyme-linked immunosorbent assays were used to measure protein levels in the arthropods. Among the 16 taxa collected in the field, Cry1Ac1 was detected in the bodies of 10. Concentrations were higher in lepidopteran larvae than in the other taxa. In particular, we found a significant correlation between Cry1Ac1 protein levels in cabbage leaves and in Pieris rapae and Mamestra brassicae. Furthermore, this protein was detected in five out of nine taxa of predators (spiders and coleopterans) and parasitoids. These results will be useful as we identify the arthropods that are directly or indirectly exposed to Bt toxin within the food web and the degree to which they are exposed during the cultivation of Bt cabbage.

Genetically modified (GM) plants have potential benefits in modern agriculture such as increasing yields and decreasing the use of chemicals. However, GM plants may cause unintended consequences of gene flow to indigenous plants or microbes and the detrimental effects on non-target organisms such as soil microbial communities. To investigate these possible unintended effects of GM plants, we conducted a 3-year field study using the transgenic pepper line H-15 carrying the coat protein gene (CMVP0-CP) of Cucumber mosaic virus (CMV) pathotype CMVP0, which is resistant to two CMV pathotypes CMVP0 and CMVP1, to assess its effects on soil fungal populations and communities. This study was conducted in two different fields using the plate counting method on selective media as well as polymerase chain reaction (PCR) and denaturing gradient gel electrophoresis analyses. In addition, we examined potential gene flow of CMVP0-CP from the transgenic line H-15 into the soil environment, including microflora, in the tested fields. We found little differences in the influence of the line H-15 compared with the inbred line P2377 (non-transgenic control) on the populations of fungal groups (total fungi, Aspergillus and Penicillium spp., Fusarium spp., and Trichoderma spp.), distribution of fungal genera, and fungal communities. Further, coat protein gene (CMVP0-CP) (PCR detection limit in soil = 1-10 pg) of the transgenic line was not detected in the soil samples of the tested fields. Taken together, the transgenic pepper line had no distinct impact on fungal populations and communities with no detection of CMVP0-CP gene flow from the transgenic line into the soil environment of the fields. Thus, the transgenic line H-15 might be little significantly harmful from the perspective of preserving soil fungal diversity; it could be a potential candidate for developing a commercial cultivar. (C) 2016 Elsevier B.V. All rights reserved.

Both herbivores that consume transgenic crops and their predators can be exposed to insecticidal proteins expressed in those crops. We conducted a tritrophic bioassay to evaluate the ecotoxicological impacts that Bt cabbage (Brassica oleracea var. capitata) expressing Cry1Ac1 protein might have on the wolf spider (Pardosa astrigera), a non-target generalist predator. Enzyme-Linked Immunosorbent Assays indicated that protein levels were 4.61 ng g(-1) dry weight in fruit flies (Drosophila melanogaster) fed with the transgenic cabbage and 1.86 ng g(-1) dry weight in the wolf spiders that preyed upon them. We also compared the life history traits of spiders collected from Bt versus non-Bt cabbage and found no significant differences in their growth, survival, and developmental rates. Because Bt cabbage did not affect the growth of fruit flies, we conclude that any indirect effects that this crop had on the wolf spider were probably not mediated by prey quality. Therefore, exposure to Cry1Ac1 protein when feeding upon prey containing that substance from transgenic cabbage has only a negligible influence on those non-target predatory spiders.

After analyzing tomato plants transformed with GalUR gene for their ascorbic acid contents, it was found that some transgenic lines contained higher levels of ascorbic acid compared to control plants. In the present study, callus induction rate was 50.2 % in the explant and shoot regeneration rate was 51.5 % from the callus with transformation efficiency of 3.0 %. Based on PCR and Southern blot analysis, three independent transformants containing the insert gene were selected. Phenotypic traits of these transgenic progeny were similar to those of control tomatoes. Tomatoes (H15) with high fruit ascorbic acid contents were selected for next generation (GalUR T-3) analysis. Transgenic tomatoes with increased ascorbic acid contents were found to be more tolerant to abiotic stresses induced by viologen, NaCl, or mannitol than non-transformed plants. In leaf disc senescence assay, the tolerance of these transgenic plants was better than control plants because they could retain higher chlorophyll contents. Under salt stress of less than 200 mM NaCl, these transgenic plants survived. However, control plants were unable to survive such high salt stress. Ascorbic acid contents in the transgenic plants were inversely correlated with MDA contents, especially under salt stress conditions. The GalUR gene was expressed in H15 tomatoes, but not in control plants. Higher expression levels of antioxidant genes (APX and CAT) were also found in these transgenic plants compared to that in the control plants. However, no detectable difference in SOD expression was found between transgenic plants and control plants. Results from this study suggest that the increase in ascorbic acid contents in plants could up-regulate the antioxidant system to enhance the tolerance of transgenic tomato plants to various abiotic stresses.

Under field conditions, we investigated how transgenic Bt cabbage expressing the insecticidal Cry1Ac1 protein affects two target Lepidoptera species—Plutella xylostella (Plutellidae) and Pieris rapae (Pieridae)—as well as the structure of the local non-target arthropod community. When exposed to Bt cabbage Line C30, both species were significantly less abundant than when in the presence of the non-transgenic control. Transgenic line C24 had no apparent influence on those target populations. Multivariate analyses (PerMANOVA and NMDS) showed that composition of the non-target community was affected by sampling date but not by cabbage genotype. These results suggest that transgenic cabbage expressing Cry1Ac1 protein can be effective in controlling P. xylostella and P. rapae in the field and that its cultivation does not adversely affect non-target arthropods.

Understanding the gene flow from genetically modified (GM) crops to conventional crops is important to prevent and mitigate seed contamination caused by pollen-mediated gene flow. We conducted a field test to investigate the gene flow from diamondback moth resistant GM cabbage (Brassica oleracea var. capitata) containing cry1Ac1 gene, to a non-GM control line AD126. GM and non-GM cabbage plants were cultivated in the field and pollinated using Bombus terrestris under the nets during the flowering periods. After seeds were collected from non-GM plants, hybrids between them and the GM cabbages were screened by multiplex PCR targeting cry1Ac1 gene. Out of 878 germinated seedlings, 168 hybrids were found and the average gene flow frequency was 19.7%. Because cabbage is mainly pollinated by insect pollinators, large-scale field tests are needed to study gene flow of GM cabbage.

해충저항성 유전자변형 작물은 대상 해충에 대한 방제 효과를 증진시킬 수 있는 효과적인 수단이 될 수 있지만, 생태계 내의 다른 비표적 생물에 부정적인 영향을 줄 가능성 또한 있기 때문에 상업적인 재배에 앞서 이에 대한 충분한 연구가 이루어져야 한다. 본 연구에서는 배추좀나방내성 유전자변형 Bt (Cry1Ac1)양배추가 비표적 곤충인 복숭아혹진딧물에 미치는 영향을 알아보기 위한 기초적인 실험으로서 기주선택실험 및 성장실험을 실시하였다. 기주선택실험에서, 두 개체씩의 Bt 양배추와 모본 양배추에 각각 10마리의 유시 성충(alate virginoparae)을 접종하고, 사흘간 생식(reproduction) 기회를 제공한 후 생산한 약충의 마리수를 측정한 결과, Bt 양배추와 모본 양배추에 낳은 약충의 수가 각각 $21.9{\pm}1.8$와 $22.5{\pm}2.2$로 비슷하였다. 성장실험에서, 갓 태어난 무시 약충(apterous virginoparae)을 Bt 양배추 및 모본 양배추를 기주로 하여 키웠을 때, 성충이 될 때까지 걸린 시간은 Bt 양배추와 모본 양배추에서 각각 $5.8{\pm}0.2$, $5.9{\pm}0.1$ 일(day)로 통계적으로 유의한 차이가 없었고, 개체군 생장률(rm) 또한 $0.7{\pm}0.1$, $0.8{\pm}0.1$로 유사하였다. 이상의 결과들로 볼때 Bt 양배추가 복숭아혹진딧물에 미치는 영향이 없거나 미미함을 알 수 있으나, 다양한 관점에서의 보완 연구가 더 필요하다. Transgenic crops that produce insecticidal toxins have a great potential for controlling target pest insects, but there is a growing concern about unintended influences on non-target species. In the present study, the preferences and performance of non-target green peach aphid, Myzus persicae (Sulzer), on transgenic cabbages (Brassica oleracea) that produce Bt toxin (Cry1Ac1) and untransformed control plants were investigated as a part of risk assessment. In a free-choice situation, the number of nymphs larviposited by 10 winged adults over 3 days was $21.9{\pm}1.8$ and $22.5{\pm}2.2$ on transgenic and the control plants, respectively, indicating that the aphids did not discriminate between the two types of plants. In a performance assay, the development time (D) and intrinsic rate of increase (rm) of wingless aphids reared on transgenic and control plants were also similar (D, $5.8{\pm}0.2$ and $5.9{\pm}0.1$ (days) and rm, $0.7{\pm}0.1$ and $0.8{\pm}0.1$, for transgenic and control plants, respectively). These results suggest that M. persicae is not significantly affected by transgenic Bt cabbage.

Pepper is a recalcitrant plant for Agrobacterium-mediated genetic transformation. Several obstacles to genetic transformation remain such as extremely low transformation rates; the choice of correct genotype is critical; and there is a high frequency of false positives due to direct shoot formation. Here, we report a useful protocol with a suitable selection method. The most important aspect of the pepper transformation protocol is selecting shoots growing from the callus, which is referred to as callus-mediated shoot formation. This protocol is a reproducible and reliable system for pepper transformation.

Chili veinal mottle virus (ChiVMV) is one of the most destructive pepper pathogens in Asia. Development of ChiVMV-resistant cultivars is necessary to control ChiVMV infection on pepper farms. However, sources of variation for ChiVMV resistance have not been identified and only a recessive resistance gene has been identified. We initially screened 30 pepper lines from several countries using inoculation tests to further examine inheritance of ChiVMV resistance, to establish a relevant breeding program, and to develop a new resistant line. Here, we report a new genetically dominant source of resistance to ChiVMV in pepper. Secondly, we found two amplified fragment length polymorphisms linked to dominant resistance and converted them into high-resolution melting markers, which were located on chromosome 6. Furthermore, we obtained a cleaved amplified polymorphic sequence marker that was closer to the ChiVMV resistance locus using comparative mapping. The newly discovered marker, related to a single dominant gene, will help researchers develop a new ChiVMV-resistant pepper cultivar.

‘Titi Chal’ with elite characteristics and high fruit yield was developed from a cross between ‘MS-BGT’ and ‘NHMF’ in 2009. The cultivar has multiple disease resistances against Tobacco mosaic virus, Tomato yellow leaf curl virus, Tomato spotted wilt virus and Leaf mold disease. The early and medium-maturing variety can be transplanted on July to September. The cherry tomato presents longer shelf-life due to higher fruit firmness and also has high eating quality with 9~10 brix of sugar content. The fruit shape is red plum type with an average fruit weight of 23~25 g. The ‘Titi Chal’ is more reliable to be cultivated because of medium-strong plant-vigor and low occurrence of creased stem.

TY Altorang' was developed from a cross between 'MS-CDE ' and 'VFR' in 2009.The cultivar has multi-disease resistances against Tobacco mosaic virus(TMV), Tomato yellow leaf curl virus(TYLCV), Fusarium crown and root rot and Leaf mold disease.Additionally, this cultivar produces larger fruit set resulting in higher fruit harvest.Fruit shape is fair and uniformed ovate type with an average fruit weight of 225 g as a beef-steak type tomato, and the fruit presents longer shelf-life due to higher fruit firmness.The early and medium-maturing variety can be transplanted in February to March, or July to September.Medium-strong plant-vigor and low occurrence of creased stem allow, relativey easy cultivation.