Impacts of Bt rice on non-target organisms assessed by the hazard quotient (HQ).

The potential risk of Bt (Bacillus thuringiensis) crops on non-target organisms (NTOs) has drawn a lot of public concerns. Despite a series of risk assessments of Bt crops on NTOs has been conducted, a quantitative approach which could support a precise judgment of their safety is required. In the present work, hazard quotient (HQ) was applied in the safety evaluation of three Bt rice events (Cry1Ab, Cry1C and Cry2Aa rice) on NTOs. Eight NTOs in different functional guilds associated with Bt rice were selected to conduct the tests. The results showed that the HQs of three Bt rice events for eight NTOs were all below the trigger value 1, while the HQ of Cry1Ab rice for one target pest Chilo suppressalis was three times higher than 1. Our results assured the reliability of the HQ and indicated that the three Bt rice events would pose no risks to the eight NTOs. Further testing of three Bt proteins on biological parameters of one NTO Nasonia virtipennis under no observed adverse effect concentration (NOAEC) confirmed the robustness of HQ assessment. We recommend that the HQ could be applied in tier-1 risk assessments of Bt crops on NTOs as a reference data standard, which would provide more clear and credible safety information of transgenic crops for the public and policy makers.

Differential impact of Bt-transgenic rice plantings on bacterial community in three niches over consecutive years.

Transgenic-Bacillus thuringiensis (Bt) crops express insecticidal proteins, which can accumulate in plants and soil where they may influence microbial populations. The impact of Bt crops on bacterial communities has only been assessed under short-term, and results have been contradictory. Here, we analyzed the bacterial communities in three niches, rhizosphere soil (RS), root endosphere (RE) and leaf endosphere (LE), of three Bt rice and their non-Bt parental lines for three consecutive years by high-throughput sequencing. In principal coordinate analysis (PCoA) and PERMANOVA (Adonis) analysis, operational taxonomic units (OTUs) were clustered primarily by niche type and differed significantly in the RE and LE but not in the RS between each of three Bt lines compared with the non-Bt rice line, and not in each respective niche among the three Bt rice lines. The bacterial communities in the RS of different rice lines over the 3 years were clustered mainly by year rather than by lines. The differential bacterial taxa among the lines did not overlap between years, presumably because Cry proteins are rapidly degraded in the soil. A network analysis of RS bacterial communities showed that the network complexity and density for the three Bt rice lines did not decrease compared with those for the non-Bt line. In conclusion, our results demonstrated that bacterial communities differed significantly in RE and LE between Bt and non-Bt rice lines, but the differences were mild and transient, and had no adverse impact on RS over the 3 years. This study provides favorable evidence in support of the commercialization of Bt rice.

Insect-Resistant Genetically Engineered Crops in China: Development, Application, and Prospects for Use.

With 20% of the world's population but just 7% of the arable land, China has invested heavily in crop biotechnology to increase agricultural productivity. We examine research on insect-resistant genetically engineered (IRGE) crops in China, including strategies to promote their sustainable use. IRGE cotton, rice, and corn lines have been developed and proven efficacious for controlling lepidopteran crop pests. Ecological impact studies have demonstrated conservation of natural enemies of crop pests and halo suppression of crop-pest populations on a local scale. Economic, social, and human health effects are largely positive and, in the case of Bt cotton, have proven sustainable over 20 years of commercial production. Wider adoption of IRGE crops in China is constrained by relatively limited innovation capacity, public misperception, and regulatory inaction, suggesting the need for further financial investment in innovation and greater scientific engagement with the public. The Chinese experience with Bt cotton might inform adoption of other Bt crops in China and other developing countries.

Safety assessment of transgenic Cry2Aa rice to a generalist predator, Paederus fuscipes Curtis (Coleoptera: Staphylinidae).

The insecticidal crystal proteins of Cry2A family from Bacillus thuringiensis (Bt) are important candidate proteins expressed in gene pyramiding Bt crops. A transgenic rice line (T2A-1) harboring a synthetic Cry2A* (Cry2Aa) gene showed effective resistance to some lepidopteran rice pests. As a generalist predator in rice ecosystems, the rove beetle (Paederus fuscipes) can prey on many rice insect pests such as planthoppers. Considering the possible exposure of Cry2Aa to P. fuscipes through tritrophic food chain, it is necessary to assess the potential risks of T2A-1 rice to this predator. In this study, a tritrophic experiment was conducted to assess the prey-mediated effects of Cry2Aa on P. fuscipes through the T2A-1 rice-Nilaparvata lugens-P. fuscipes food chain. After preying on N. lugens nymphs reared on T2A-1, no accumulated Cry2Aa could be detected in P. fuscipes adults, despite Cry2Aa being detected in N. lugens. In addition, no harmful effects were detected on the life table parameters of P. fuscipes in this tritrophic chain. Additionally, direct exposure to a high dose of purified Cry2Aa protein, representing the worst case scenario, showed no significant adverse effects on the development of P. fuscipes. These results showed that transgenic Cry2Aa rice had no harmful effects on P. fuscipes.

Effects of Cry1Ab-expressing Bt rice straw return on juvenile and adult Eisenia fetida.

A 90 day experiment was conducted in the laboratory to investigate the potential effects of transgenic Cry1Ab-expressing rice (Bacillus thuringiensis (Bt) rice: T775 and its F1 hybrid) straw return on earthworm Eisenia fetida, compared to non-Bt rice (TYHZ) straw. Juvenile E. fetida could survive, grow up, mature and reproduce offspring well in a Bt rice treated test during the whole experiment. The significantly higher relative growth rate (RGR) was found in earthworms from Bt rice treatment than from non-Bt rice treatment on the 7th day. The period of sexual maturity for earthworms from Bt rice treatments was shortened significantly, compared to non-Bt rice treatments. Adult E. fetida survived with weight loss under Bt rice treatments. On the 7th and 15th day, earthworm RGR decreased and glutathione peroxidase (GSH-PX) activity increased under Bt rice straw treatments. Significantly fewer offspring were produced by earthworms from Bt rice than non-Bt rice treatments on the 60th and 75th day. Enzyme-linked immunosorbent assay (ELISA) determined a sharp decrease of Cry1Ab in straw mixed soil along with the experimental time, regardless of juvenile or adult earthworm treatments. Cry1Ab concentration in the earthworms from the juvenile group was significantly higher than those from the adult group. Bt rice straw return had significant effects on soil nutrients, especially on the content of total and available phosphorus. In view of two bioassays, Bt rice (T775 and its F1 hybrid) straw return presented different effects on E. fetida from the juvenile (no deleterious effect) and adult (a little negative effect) groups, that were not directly related to Cry1Ab presence and nutrient differences among the three rice variety treatments.

Degradation of transgenic Bt-rice straw incorporated with two different paddy soils.

Transgenic Bt-rice is rice that has been genetically modified to produce insecticidal proteins (Cry1Ab/Ac) within the plant. Rice straw is incorporated into paddy soils after harvest for fertilization or to improve the soil structure. The incorporation of straw from transgenic Bt-rice may pose risks to the paddy soil system. The decomposition of Bt-rice straw and degradation of Cry1Ab/Ac proteins from the straw were investigated under laboratory conditions. In addition, effects of the incorporation with chopped rice straw on microbial communities in differently textured paddy soils were studied. The results indicated that the incorporation of straw from transgenic Bt-rice might have a slight influence on soil respiration and CH4 emissions in two paddy soils, i.e. the Silt Loam soil and the Silty Clay soil. Differences were also observed in the cumulative emissions of CO2 between the two amended paddy soils in addition to the well-known increase in emissions of both CO2 and CH4 due to straw incorporation. The Cry1Ab/Ac proteins from straw of transgenic Bt-rice were degraded in paddy soils. The rate of decline in the concentration of Cry1Ab/Ac proteins was different in the two soils. After 29 d of incubation, 61% and 42% of initial Cry1Ab/Ac proteins were detected in the silt loam and silty clay, respectively. As a result of the presence of the rice straw, the abundance of bacteria, archaea, and total cells were increased in two soils. The numbers of bacteria and total cells were 6.4% and 11.5% higher in the silt loam amended with straw of Bt-rice than non-Bt-rice, respectively. The silty clay displayed a similar trend as the silt loam.

Bt rice plants may protect neighbouring non-Bt rice plants against the striped stem borer, Chilo suppressalis.

The area planted with insect-resistant genetically engineered crops expressing Bacillus thuringiensis (Bt) genes has greatly increased in many areas of the world. Given the nearby presence of non-Bt crops (including those planted as refuges) and non-crop habitats, pests targeted by the Bt trait have a choice between Bt and non-Bt crops or weeds, and their host preference may greatly affect insect management and management of pest resistance to Bt proteins. In this study, we examined the oviposition preference of the target pest of Bt rice, Chilo suppressalis, for Bt versus non-Bt rice plants as influenced by previous damage caused by C. suppressalis larvae. The results showed that C. suppressalis females had no oviposition preference for undamaged Bt or non-Bt plants but were repelled by conspecific-damaged plants whether Bt or non-Bt Consequently, C. suppressalis egg masses were more numerous on Bt plants than on neighbouring non-Bt plants both in greenhouse and in field experiments due to the significantly greater caterpillar damage on non-Bt plants. We also found evidence of poorer performance of C. suppressalis larvae on conspecific-damaged rice plants when compared with undamaged plants. GC-MS analyses showed that larval damage induced the release of volatiles that repelled mated C. suppressalis females in wind tunnel experiments. These findings suggest that Bt rice could act as a dead-end trap crop for C. suppressalis and thereby protect adjacent non-Bt rice plants. The results also indicate that the oviposition behaviour of target pest females should be considered in the development of Bt resistance management strategies.

Effects of transgenic Bt rice on the active rhizospheric methanogenic archaeal community as revealed by DNA-based stable isotope probing.

AIMS: This study aimed to investigate the influence of planting Cry1Ab/Cry1Ac gene expressing rice (Bt rice) on rhizospheric active methanogenic archaeal communities. METHODS AND RESULTS: The nontransgenic parental line was used as the control (Ck rice). DNA-based stable isotope probing (DNA-SIP) technology traced the rhizospheric active methanogens at the tillering stage. The results revealed significantly lower CH4 emission flux from Bt soil than that from Ck soil during the whole growth period. The active methanogenic community composition remained stable. The RC-I lineage (77·9-79·8%) and Methanosaetaceae (13·9-15·1%) were the predominant active methanogens in Bt and Ck rice rhizospheres. However, the abundance of functionally active methanogens in the Bt rice rhizosphere was significantly reduced. Lower levels of root exudates (that included carbohydrate and organic acids) from Bt rice were also detected at the tillering stage. CONCLUSION: This study found that the genetic modification of rice reduced the potential methanogenic substrates came from plant-derived root exudates, which represented an important factor in reducing CH4 generation and active methanogenic archaeal abundance in Bt rhizosphere soil. SIGNIFICANCE AND IMPACT OF THE STUDY: The effect of genetically modified (GM) insect-resistant crops on soil micro-organisms has become an issue of public concern, especially the indirect effect of plant metabolisms caused by the insertion of foreign genes. Methanogenesis, which is regarded as a critical ecological process in paddy soil, is influenced by plant root exudates; these are mainly derived from photosynthesis. The variations in root exudates across the Bt and Ck rice suggested the indirect influence of foreign gene insertion. DNA-SIP successfully traced the active methanogenic archaeal populations assimilating 13 C-labelled photosynthetic carbon and found a strong influence of planting Bt rice on active methanogens. As a consequence, we proposed that analysis of functionally active micro-organisms is more suitable for monitoring and predicting the environmental influence of GM plants.

Probing active microbes involved in Bt-containing rice straw decomposition.

Transgenic Bacillus thuringiensis (Bt) rice extends significant protection against insect pests and meets the increasing demands for food and energy. Many studies have been conducted investigating the impacts of Bt rice to the agricultural ecosystem, but much less attention has been given to efforts attempting to determine how the presence of Bt rice influences and shapes the microbial community, especially the active microbes. Stable isotope probing and high-throughput sequencing were employed to explore the active microbes involved in Bt-containing straw decomposition. Compared to its near isoline, the Bt straw contained higher contents of total N, total P, total K, lignin, cellulose, and Cry1Ab toxin protein. These chemical differences did not affect the decomposition rate but significantly changed the active microbial decomposer communities. During the decomposition of Bt-containing straw, fungi were more affected than bacteria. Agromyces, Terrabacter, Microbacterium, Glycomyces, and Kribbella were the most representative unique (existed only in the Bt treatments and appeared at the early stage) bacterial genera, and Trichoderma was the most representative unique fungal genus in the Bt straw decomposition. By using similarity index calculation and function prediction, the significant differences between Bt straw and non-Bt straw treatments were found to be transient for both microbial taxa and functional traits. These results suggested that Bt rice has a significant but transient impact on soil microbes in terms of microbial straw decomposition.

Developmental immunotoxicity is not associated with the consumption of transgenic Bt rice TT51 in rats.

TT51 is a transgenic strain of Bt rice generated by fusing a synthetic CryAb/Ac gene into MingHui rice. In this study, rats from F0, F1, and F2 generations were fed a diet with 60% TT51 rice, MingHui rice, or nominal-origin rice. The study focused on developmental immunotoxicity in F1 and F2 offspring after long-term consumption of TT51. A wide range of immunological parameters was monitored in this two-generation study on reproductive toxicity. The experiments were performed on F1 and F2 offspring at postnatal days 21 and 42. No adverse clinical effects were observed in any of the experimental groups. In addition, histopathology observations and immunotoxicity tests, including hematological indicators, spleen lymphocyte subsets, natural killer cell activity, lymphoproliferative response, and plaque-forming cell assay, revealed no significant difference between the groups. These results indicated that developmental immunotoxicity was not associated with a diet of transgenic Bt rice TT51, compared to the parental MingHui rice.