Transgenic Bt Rice Research Articles

Bt-Modified Transgenic Rice May Shift the Composition and Diversity of Rhizosphere Microbiota.

Plants significantly shape root-associated microbiota, making rhizosphere microbes useful environmental indicator organisms for safety assessment. Here, we report the pyrosequencing of the bacterial 16S ribosomal RNA in rhizosphere soil samples collected from transgenic cry1Ab/cry1Ac Bt rice Huahui No. 1 (GM crop) and its parental counterpart, Minghui63. We identified a total of 2579 quantifiable bacterial operational taxonomic units (OTUs). Many treatment-enriched microbial OTUs were identified, including 14 NonGM-enriched OTUs and 10 GM-enriched OTUs. OTUs belonging to the phyla Proteobacteria, Actinobacteria, Acidobacteria, Firmicutes, Nitrospirae, Chlorobi and GN04 were identified as statistically different in abundance between GM and the other two treatments. Compared with the different impacts of different rice varieties on microbiota, the impact of rice planting on microbiota is more obvious. Furthermore, Huahui No. 1 transgenic Bt rice had a greater impact on the rhizosphere bacterial communities than Minghui63. Early developmental stages of the transgenic Bt rice had a significant impact on many Bacillaceae communities. Soil chemical properties were not significantly altered by the presence of transgenic Bt rice. The peak concentration level of Bt protein products was detected during the seedling stage of transgenic Bt rice, which may be an intriguing factor for bacterial diversity variations. Based on these findings, we conclude that transgenic Bt rice has a significant impact on root-associated bacteria. This information may be leveraged in future environmental safety assessments of transgenic Bt rice varieties.

Knockdown of MAPK p38-linked genes increases the susceptibility of Chilo suppressalis larvae to various transgenic Bt rice lines

Bacillus thuringiensis (Bt) toxins have provided exceptional control of agricultural insect pests, however, over reliance on the proteins would potentially contribute to the development of field tolerance. Developing new sustainable insect pest control methods that target the mechanisms underlying Bt tolerance can potentially support the Bt control paradigm while also providing insights into basic insect physiology. The MAPK p38 pathway is strongly associated with Bt tolerance in Chilo suppressalis, a major pest of rice. To gain insights into how this pathway impacts tolerance, high-throughput screening of C. suppressalis larval midguts initially identified eight novel target genes. Increased larval sensitivity to the transgenic cry1Ca rice strain T1C-19 was observed following RNA interference-mediated knockdown of four of the genes, Cscnc, Csgcp, Cszfp26 and CsZMYM1. Similar enhanced sensitivity to the TT51 (expressing Cry1Ab/1Ac) and T2A-1 (expressing Cry2Aa) transgenic rice lines occurred when Cszfp26 and CsZMYM1 were knocked down. All four target genes are downstream of the MAPK p38 pathway but do not participate in negative feedback loop of the pathway. These results implicate Cscnc, Csgcp, Cszfp and CsZMYM1 in the C. suppressalis transgenic cry1Ca rice tolerance mechanism regulated by MAPK p38. These findings further enhance our understanding of the MAPK p38-dependent molecular mechanisms underlying Bt tolerance in C. suppressalis and open new avenues of tolerance management to develop.

Physiological Response of the Target Stemborer Chilo suppressalis to Elevated CO2 as Reared with Transgenic Bt Rice during Different Plant Growth Stages

Physiological Response of the Target Stemborer Chilo suppressalis to Elevated CO2 as Reared with Transgenic Bt Rice during Different Plant Growth Stages

Analysis of the Unintended Effects of the Bacillus thuringiensis Insecticidal Protein in Genetically Modified Rice Using Untargeted Transcriptomics

The safety and unintended effects of genetically modified (GM) crops have been the focus of public attention. Transcriptome analysis is a powerful tool to assess the potential impact of genetic modification on plant genomes. In this study, three transgenic (KMD, KF6, and TT51-1) and three non-transgenic (XS11, MH86, and MH63) rice varieties were assessed at the genomic and protein levels. The results of polymerase chain reaction (PCR) and Cry1Ab/1Ac speed test strips showed that the Bt gene was successfully expressed in transgenic rice. The results of RNA-seq analysis to analyze the unintended effects of transgenic Bt rice showed fewer differentially expressed genes (DEGs) between the transgenic and non-transgenic rice varieties than among the different varieties. Meanwhile, the results of principal component analysis and cluster analysis found no significant genetic variation between the transgenic and non-transgenic rice varieties, except for the presence of Bt in transgenic rice. There were only two co-upregulated DEGs and no co-downregulated DEGs among three comparison groups. Although there were various DEGs among the groups, the two co-upregulated DEGs were not related to any significantly enriched gene ontology (GO) term or Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway, indicating that the differences among the subgroups were more likely caused by complex environmental or genetic factors, rather than unintended effects due to Bt expression. This study provides useful information to further explore the unexpected effects and safety of GM rice.

Application and Development of Bt Insect Resistance Genes in Rice Breeding

As pests are an important factor in reducing crop yields, pest control is an important measure in preventing reductions in crop yields. With the aim of ending the use of chemical pesticides, biological control and genetically modified methods are now considered more reasonable pest control strategies. The bacterium Bacillus thuringiensis (Bt) can produce crystal proteins that have specific toxicity to lepidopteran insects, and so it has been applied as a microbial insecticide in the control of crop pests for several decades. With the development of plant genetic engineering, Bt genes encoding insecticidal crystal protein have been introduced into many crop species for pest control. This article indicates that, after years of experiments and research, Bt transgenic rice is close to becoming a commercial insect-resistant rice, and many studies have shown that transgenic rice has pronounced abilities in the control of pests such as yellow stem borers (Scirpophaga incertulas, YSB), striped stem borers (Chilo suppressalis, SSB), and rice leaf rollers (Cnaphalocrocis medinalis, RLR); moreover, it does not obviously differ from non-transgenic rice in terms of safety. This paper suggests that transgenic Bt rice has application potential and commercial value.

Monitoring reduced accumulation and risk evaluation of Cry1Ca during food chain transfer from genetic engineered plant to endoparasitoid.

The potential risk of insecticidal proteins produced by genetically engineered (GE) plants to non-target organisms have long been an ecotoxicological concern. Apanteles chilonis, an important endoparasitoid of rice pest Chilo suppressalis, is potentially exposed to Bacillus thuringiensis (Bt) endotoxins through a food chain of transgenic Bt rice - C. suppressalis - A. chilonis, thus, a rigorous risk assessment is urgently needed. Here we combined a tri-trophic bioassay system with high dose exposure approach using C. suppressalis hemolymph as the carrier of insecticidal protein to evaluate the biosafety of Cry1Ca to A. chilonis. Cry1Ca protein could be transmitted and retained along the food chain and remains bioactive in the hemolymph of C. suppressalis during the pre-adult duration of A. chilonis. No significant differences in pre-adult period, male and female longevity, adult fecundity and weight, emergence rate, and sex ratio were observed when A. chilonis parasitized C. suppressalis feeding on cry1Ca rice compared to control treatment. However, the pupal period and weight were significantly prolonged and decreased. When A. chilonis parasitized on C. suppressalis injected with a high dosage of Cry1Ca protein, no adverse effects on the life-history parameters, peroxidase (POD), superoxide dismutase (SOD), and glutathione reductase (GR) of A. chilonis were observed, demonstrating that the host quality mediates adverse effects during the food chain. We confirmed that Cry1Ca posed no ecological risk to the non-target endoparasitoid A. chilonis. This study may serve as an example for future risk assessment of transgenic crops to non-target endoparasitoids. This article is protected by copyright. All rights reserved.

Effect of Bt traits on transgenic rice’s growth and weed competitiveness

Transgene escape could lead to genetically modified rice establishing wild populations in the natural environment and competing for survival space with weeds. However, whether the expression of the Bacillus thuringiensis (Bt) gene in rice will alter the relationship between transgene plants and weeds and induce undesirable environmental consequences are poorly understood. Thus, field experiments were conducted to investigate the weed competitiveness and assess the ecological risk of transgenic Bt rice under herbicide-free and lepidopterous pest-controlled environments. Results showed that weed–rice competition in the direct-sowing (DS) field was earlier and more severe than that in the transplanting (TP) field, which resulted in a significant decrease in biomass and yield in DS. However, conventional Bt and non-Bt rice yield was not significantly different. The weed number, weed coverage ratio, and weed diversity of conventional Bt rice were significantly higher than those of non-Bt rice at the early growth and mature stages, especially in DS plots, suggesting that Bt traits did not increase the weed competitiveness of transgenic rice and had no negative effect on weed diversity. Grain yield and weed number varied between different hybrid rice lines, but those differences were insignificant between Bt and non-Bt rice. The number of insects increased with the increase of weeds in hybrid rice plots, whereas the insect number and diversity did not display a significant difference between Bt and non-Bt rice. Therefore, the ecological risk of transgenic Bt rice is comparable to non-Bt rice.

Delta and jagged are candidate target genes of RNAi biopesticides for the control of Nilaparvata lugens.

The brown planthopper (BPH; Nilaparvata lugens) is an important pest in rice cultivation, and chemical pesticide over-use and ineffectiveness of existing Bt transgenic rice against piercing-sucking insects make novel control methods necessary. RNA interference (RNAi) biopesticide is a new type of product with high efficiency and specificity and are simple to use. The Notch signaling pathway has extensive and important physiological functions and plays a key role in the development of insects. In this study, two key ligand genes of the Notch signaling pathway, delta (dl) and jagged (jag), were selected and their lethal effects and functional analysis were systematically evaluated using a stable short-winged population (Brachypterous strain) and a long-winged population (Macropterous strain) of BPHs. The full-length coding sequences of Nldl and Nljag comprised 1,863 and 3,837 base pairs, encoding 620 and 1,278 amino acids, respectively. The nucleic acid sequences of Nldl and Nljag were identical between the two strains. The expression levels of Nldl and Nljag were relatively high in the head of the nymphs, followed by those in the abdomen. Through RNAi treatment, we found that injection of BPH nymphs of both strains with dsNldl (10-50ng/nymph) or dsNljag (100ng/nymph) produced lethal or teratogenic effects. dsRNA treatment showed excellent inhibitory effects on the expression of target genes on days 1 and 5, suggesting that RNAi rapidly exhibits effects which persist for long periods of time in BPHs. Taken together, our results confirm the potential of Nldl and Nljag as target genes of RNAi biopesticides, and we propose optimized dosages for the control of BPHs.

The long-term persistence of transgenic volunteers in wild mustard and rice populations

The undesired presence of GM plants outside of cultivation is one of main concerns for the ecological risk assessment and regulation of GM plants, and how long transgenic volunteers can persist in the nature remains unknown. We conducted two long-term coexistence experiments of Bt-transgenic insect-resistant crops in populations of their wild relatives, using Bt-transgenic oilseed rape (Brassica napus) in wild mustard (B. juncea) populations from 2012 to 2019, and Bt-transgenic rice (Oryza sativa) in wild rice (O. rufipogon and O. minuta) populations from 2013 to 2019. Transgenic oilseed rape volunteers survived only in the 2012 winter, because it is a spring variety and not be resistant to cold climate and competition from weeds. Transgenic rice was not survived because of its low competitive ability compared to wild rice, but survived five years in one population of wild rice O. minuta who could not tolerant to cold temperature. Our results indicated that transgenic volunteers can persist in the wild populations under natural conditions, but the fate of transgenic volunteers in the nature depends on whether the growth environment is favorable, and they could not disperse over its niche or “tolerance zone”.

Evaluation of Chilo suppressalis resistance and analysis of CRY1C expression in transgenic rice

AbstractThe insect resistance of Bacillus thuringiensis (Bt) transgenic rice (Oryza sativa L.) is mainly determined by the transcription of the CRY1C gene and the translation process of the Cry1C protein. With this in mind, we analyzed CRY1C expression and Cry1C protein content in a transgenic Bt insect‐resistant restorer line and its F1 hybrids, evaluated the resistance to Chilo suppressalis of various lines of rice, and determined the 50% lethal concentration (LC50) of Cry1C for C. suppressalis. In four transgenic rice lines, the relative expression of CRY1C was highest at the heading stage in most tissues. Among different tissues from the same developmental stage, CRY1C expression was highest in leaves, followed by stems and panicles. The relative expression of CRY1C was higher in the parent restorer line than in the F1 hybrids. The LC50 of the Cry1C protein for C. suppressalis was 4.016 μg g−1, and the Cry1C protein expression level of transgenic insect‐resistant rice exceeded this threshold at the heading stages and in stems. We next analyzed the lethality of each strain toward C. suppressalis. After 48 h, the mortality rate of second‐instar C. suppressalis larvae feeding on transgenic stem tissue was higher at the heading stage than at the tiller stage and varied from 66.7 to 91.7%. By documenting the temporal and spatial expression of CRY1C and evaluating C. suppressalis resistance to Bt transgenic rice, the risk of pests and diseases during the rice production process can be greatly reduced.

Accumulation of Cry proteins in soil released from Bt rice after planting for multiple years

Cry protein residue and accumulation in soil are two important components of the environmental safety assessment for the plantation of transgenic Bt crops. Several Bt rice lines with good commercial prospects have been developed in China, but it is unclear whether Cry proteins will accumulate in soils after multiple years of Bt rice cultivation. We planted the transgenic Bt rice lines cry1Ab/1Ac Minghui 63 (Huahui No. 1) and cry2A Minghui 63 for 9 years in the same field. The Cry proteins in the rhizosphere soil were estimated with enzyme linked immunosorbent assay (ELISA) at tillering stage and on the 60th day after harvest in each year. The Cry protein residues during the seedling, flowering and ripening stages were estimated in the first year (2012) and the last year (2020) of the experiment. In 2012, the concentration of Cry1Ab/1Ac in the rhizosphere soil of Huahui No. 1 was 1.25, 1.77, 1.97, 1.71 and 0.30 ng·g-1 at the seedling, tillering, flowering, ripening stages and on the 60th day after harvest, respectively. In 2020, the corresponding values were 1.30, 1.69, 2.03, 1.77, and 0.43 ng·g-1. In 2012, the concentration of Cry2A in rhizosphere soil of line cry2A Minghui 63 was 0.91, 1.52, 1.53, 1.37, and 0.12 ng·g-1 at the seedling, tillering, flowering, ripening stages and on the 60th day after harvest, respectively. The corresponding values in 2020 were 0.95, 1.43, 1.61, 1.40, and 0.15 ng·g-1. Results of multi-way ANOVA showed that the effect of year was not significant, but the effects of rice variety and growth stage were significant. Our results indicated that Cry proteins could be detected in rhizosphere soil during the growth stages of Bt rice, but would be degraded by 60 d after harvest, and that the concentrations of Cry proteins in the soil would not accumulate across multiple planting years.

Trait-based comparison of transgenic Bt rice and its non-Bt counterpart in response to soil copper pollution.

Transgenic Bacillus thuringiensis (Bt) rice can provide economic and environmental benefits under the current increasing demand for food and socioeconomic pressures for sustainability. However, information about the ecological adaptation of Bt rice under nontarget environmental stress is still lacking. This study compared the adaptability of one Bt rice and its nontransgenic counterpart to soil copper (Cu) pollution in terms of agronomic and physiological traits. With Cu addition, grain yield and biomass of both cultivars were significantly decreased. Within the same Cu treatment, Bt rice exhibited higher biomass and close plant height, chlorophyll content, grain yield, and grain quality compared with non-Bt rice, except for the grain yield with a 35 mg kg-1 Cu addition with respect to which Bt rice was significantly lower by 22%. The Cu content in Bt rice was generally lower, whereas the antioxidant enzyme activity and lipid peroxidation were stronger than the non-Bt. These results demonstrated that Bt rice exhibited close adaptability but higher Cu tolerance compared with the non-Bt under Cu stress.

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.

Compositional equivalence assessment of insect-resistant genetically modified rice using multiple statistical analyses

ABSTRACT The safety of transgenic Bt rice containing bacteria-derived mCry1Ac gene from Bacillus thuringiensis (Bt) was assessed by conducting field trials at two locations for two consecutive years in South Korea, using the near-isogenic line comparator rice cultivar (‘Ilmi’, non-Bt rice) and four commercial cultivars as references. Compositional analyses included measurement of proximates, minerals, amino acids, fatty acids, vitamins, and antinutrients. Significant differences between Bt rice and non-Bt rice were detected; however, all differences were within the reference range. The statistical analyses, including analysis of % variability, analysis of similarities (ANOISM), similarity percentage (SIMPER) analysis, and permutational multivariate analysis of variance (PERMANOVA) were performed to study factors contributing to compositional variability. The multivariate analyses revealed that environmental factors more influenced rice components’ variability than by genetic factors. This approach was shown to be a powerful method to provide meaningful evaluations between Bt rice and its comparators. In this study, Bt rice was proved to be compositionally equivalent to conventional rice varieties through multiple statistical methods.

Transcriptomic and proteomic responses to brown plant hopper (Nilaparvata lugens) in cultivated and Bt-transgenic rice (Oryza sativa) and wild rice (O. rufipogon)

Strategies are still employed to reduce insect damage in crop production, including conventional breeding with wild germplasm resources and transgenic technology with foreign genes' insertion. Cultivated and Bt-transgenic rice (Oryza sativa) and two ecotypes of wild rice (O. rufipogon) were treated by a 72 h feeding of brown plant hopper (Nilaparvata lugens). Under the feeding of N. lugens, compared with the cultivated rice (568 and 4), more differentially expressed genes (DEGs) and differentially accumulated proteins (DAPs) were identified in transgenic rice (2098 and 11) and two wild ecotypes (1990, 39 and 1932, 25, respectively). The iTRAQ analysis showed 79 DAPs and confirmed the results of RNA-seq, which showed the least GO terms and KEGG pathways responding to herbivory in the cultivated rice. DAPs significantly enriched two GO terms that are related with Bph14 and Bph33 genes in rice. Most of DEGs and DAPs were related to plant biological processes of plant-pathogen interaction and plant hormone signal transduction, and hormone signaling and transcription factors regulate the immune response of rice to BPH. Our results demonstrated the similarity in the wild rice and Bt-transgenic rice for their transcriptomic and proteomic response to herbivory, while cultivated rice lacked enough pathways in response to herbivory. Statement of significance of the studyThe iTRAQ analysis and RNA-seq were employed 39 to identify differentially expressed genes (DEGs) and differentially accumulated proteins (DAPs) in seedlings of cultivated, Bt-transgenic and two wild rice ecotypes under feeding of brown plant hopper. Wild rice showed DEGs and DAPs related to biochemical pathways of plant pathogen interactions and plant hormone signal transductions, while cultivated rice lacked enough pathways in response to herbivory. Crop domestication weakened the response of plants to herbivory, while the insertion of Bt gene might promote the response of plants to herbivory. Growing environment plays an important role in regulating gene networks of plant response to herbivory. Our results highlighted the importance of conservation of crop wild species. SignificanceInsect damage is one of main factors in reducing agricultural production, and technologies and methods were employed to control insect pests in agricultural systems. Transgenic technology is developed to produce insect-resistant crops, but receive concerns on biosafety risks. Alternatively, crop wild species are important genetic resource in crop breeding to produce trait-specific varieties. Here, we investigated the molecular mechanisms of plant response to herbivory in wild, Bt-transgenic and cultivated rice, and found crop domestication weakened the response of plants to herbivory. The insertion of foreign Bt gene may promote the expression of other genes. In addition, our results showed growing environment plays an important role in regulating gene networks of plant response to herbivory. These results highlight the importance of wild species conservation, with the strategy of in situ conservation.

Elevated CO2 alters transgene methylation not only in promoterregion but also in codingregion of Bt rice under different N-fertilizer levels

The earth has been undergoing climate change, especially in recent years, driven by increasing concentration of atmospheric carbon dioxide (CO2) and rising earth-surface temperature, which could reduce N allocation to Bt toxin for transgenic Bt crops (Bt crops), but the N fertilization is considered to be an effective method to enhance the C–N balance in Bt crops in the case of elevated CO2 in future. DNA methylation not only in promoterregion but also in codingregion of transgene plays a critical role in transgene expression regulation and silencing of transgenic crops. Recent research has emphasized the risks of increased transgene silencing of Bacillus thuringiensis (Bt) rice under elevated CO2. In this study, the effects of elevated CO2 (vs. ambient CO2) on exogenous Bt toxins and transgene expression in promoterregion and codingregion of Bt rice during tillering stage (cv. HH1 expressing fused Cry1Ab/Cry1Ac) were evaluated under three nitrogen (N) fertilizer rate (1/4, 1 and 2 N levels). The aboveground and belowground biomass, and foliar Bt protein content of Bt rice were all significantly increased with the augmentation of N-fertilizer. And elevated CO2 significantly increased belowground biomass, total soluble protein content, transgene methylation levels in promoterregion (P1), and in total of promoterregion(P1) and codingregion (P2 + P3) (i.e., P1 + P2 + P3) at 1 N level, and it also increased transgene methylation levels in codingregion (P2), and in total of promoterregion and codingregion (P1 + P2 + P3) at 2 N level. In addition, elevated CO2 decreased foliar Bt protein content at 1 N level. The transgene methylation levels in promoterregion and codingregion were negatively correlated with Bt-transgene expression level. The methylation level of cytosines located at CG sites was higher than those at CHG and CHH sites in P1, P2 and P3 fragments regardless of the CO2 or N-fertilizer level. The correlation of transgene mehtylation in promoterregion with transgene expression is even stronger than that in codingregion. These data indicate that N fertilization supply will increase the Bt toxin content in transgenic Bt rice, especially under elevated CO2.

Differences in midgut transcriptomes between resistant and susceptible strains of Chilo suppressalis to Cry1C toxin

BackgroundChilo suppressalis is a widespread rice pest that poses a major threat to food security in China. This pest can develop resistance to Cry toxins from Bacillus thuringiensis (Bt), threatening the sustainable use of insect-resistant transgenic Bt rice. However, the molecular basis for the resistance mechanisms of C. suppressalis to Cry1C toxin remains unknown. This study aimed to identify genes associated with the mechanism of Cry1C resistance in C. suppressalis by comparing the midgut transcriptomic responses of resistant and susceptible C. suppressalis strains to Cry1C toxin and to provide information for insect resistance management.ResultsA C. suppressalis midgut transcriptome of 139,206 unigenes was de novo assembled from 373 million Illumina HiSeq and Roche 454 clean reads. Comparative analysis identified 5328 significantly differentially expressed unigenes (DEGs) between C. suppressalis Cry1C-resistant and -susceptible strains. DEGs encoding Bt Cry toxin receptors, aminopeptidase-P like protein, the ABC subfamily and alkaline phosphatase were downregulated, suggesting an association with C. suppressalis Cry1C resistance. Additionally, Cry1C resistance in C. suppressalis may be related to changes in the transcription levels of enzymes involved in hydrolysis, digestive, catalytic and detoxification processes.ConclusionOur study identified genes potentially involved in Cry1C resistance in C. suppressalis by comparative transcriptome analysis. The assembled and annotated transcriptome data provide valuable genomic resources for further study of the molecular mechanisms of C. suppressalis resistance to Cry toxins.

Evaluating the effects of transgenic Bt rice cultivation on soil stability.

Insecticidal crystal (Cry) proteins produced by genetically modified rice that enter the soil via pollen dispersal, plant residues, and root exudation may disturb soil health. In the present study, we assessed the influences of transgenic Bt rice (i.e., HH1 with Cry1Ab/Cry1Ac) cultivation on the dynamics of soil carbon and nutrients under field conditions during 2013-2016. Transgenic treatments (transgenic Bt rice vs. its parental line (i.e., MH63) of non-Bt rice) have no consistently significant effects on soil property, including available nitrogen, available phosphorus, available potassium, total nitrogen, and total phosphorus, while apparent seasonal changes were observed. Besides, the variations of soil nutrients in the paddy field of transgenic Bt rice did not exceed their resistance capacities, except total organic carbon (TOC; RS (resistance) = 1.51) and total potassium (TK; RS = 2.62) in 2013 and TK (RS = 1.94) in 2014. However, the TOC and soil nutrient of TK in the paddy field of transgenic Bt rice have recovered to the pre-perturbation status after harvest (RL (resilience) = 1.01, F = 0.01, P = 0.91; RL = 0.98, F = 0.34, P = 0.58; RL = 0.99, F = 1.26, P = 0.29). Moreover, the paddy yield of transgenic Bt rice was consistently higher than that of its parental line of non-Bt rice. These results suggested that the cultivation of transgenic Bt rice has no adverse impact on soil stability in terms of soil carbon and nutrients and paddy yield.

Effects of Pest Management Practices on Soil Nematode Abundance, Diversity, Metabolic Footprint and Community Composition Under Paddy Rice Fields.

The wide-scale adoption of transgenic crops has aroused public concern towards potential impacts to the ecological services of soil fauna, such as soil nematodes. However, few studies has examined whether the cultivation of transgenic rice would pose greater threats to soil nematode community and associated ecological functions than insecticides application. Moreover, what are determinants of soil nematode community in paddy fields remains unclear. During a 3-year field study, rhizosphere soil samples of transgenic-Bt rice, its counterpart non-Bt parental rice and not-Bt rice with insecticides application were taken at four times in the rice developmental cycle using a random block design with three replications for each treatment. We hypothesized that the effects of pest management practice on soil nematode abundance and metabolic footprint change with trophic group and sampling time. We also predicted there were significant differences in structure and composition of soil nematode community across the three treatments examined and sampling times. In agreement with our expectation, the effects of pest management practice on nematode abundance and metabolic footprints depend on trophic group and sampling time. However, pest management practice exerted no apparent effect on nematode diversity and community composition. Soil nutrient availability and C:N molar ratio are the primary regulating factor of soil nematode community in rice paddy fields. In conclusion, our findings implied that changes in abundance, diversity, metabolic footprints associated with the crop growth stage overweighed the application of Bt rice and insecticides. The cultivation of Bt rice Huahui-1 exerted no measurable adverse effect on soil nematode community in rhizosphere soil over 3 years of rice cropping.

Cry1Ab/Ac proteins released from subspecies of Bacillus thuringiensis (Bt) and transgenic Bt-rice in different paddy soils

ABSTRACT As Bacillus thuringiensis (Bt) are entomopathogenic bacteria for use as biological insecticide and serve as a main source for genetically modified crops, it is imperative to investigate fate of Bt-toxins (Cry-proteins) released in soil. Four differently textured paddy soils, i.e. sandy loam, loam, silt loam and silty clay, were selected to investigate production and release of Cry-proteins by two Bt-strains (subspecies thuringiensis and kurstaki) and by transgenic Bt-rice (cultivar TT51-1). After amended with Cry-proteins, soil can adsorb proteins while sorption rate was variable among different paddy soils, with the highest amount of Cry-proteins in the silty clay soil. During the period of incubation with Bt-strains, the silty clay soil showed significant accumulation of Cry-proteins. Additionally, level of Cry-protein production was different between two Bt-strains. Inactivation of indigenous microorganisms in the soil samples led to slightly higher production of Cry-proteins by Bt-strains in all four soil types. Cry-proteins were released via root exudates of transgenic Bt-rice in sterile hydroponic culture and in non-sterile soils throughout the growth of plants. Protein release significantly varied among different paddy soils, up to 34.9 ng Cry-proteins g−1 soil in clayey soil. It indicates differently textured paddy soils reflect variable sorption, persistence, and accumulation of Cry-proteins.