Discrimination of Transgenic Rice containing the Cry1Ab Protein using Terahertz Spectroscopy and Chemometrics.
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ABSTRACT:
SUBMITTER: Xu W
PROVIDER: S-EPMC4495602 | biostudies-other | 2015
REPOSITORIES: biostudies-other
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Project description:Management of resistance development of insect pests is of great importance for continued utilization of Bt crop. The high-dose/refuge and pyramid (gene stacking) strategy are commonly employed to delay the evolution of insect resistance. Due to the anticipated difficulty for deployment of mandatory refuge for transgenic crops in China, where the size of farmer is quite small, stacking of genes with different modes of action is a more feasible strategy. Here we report the development of transgenic rice expressing a fusion protein of Cry1Ab and Vip3A toxin. Analysis of trypsin proteolysis suggested that the fusion protein is equivalent to the combination of Cry1Ab and Vip3A protein. The transgenic plants expressing the fusion protein were found to be highly resistant to two major rice pests, Asiatic rice borer Chilo suppressalis (Lepidoptera: Crambidae) and rice leaf folder Cnaphalocrocis medinalis (Lepidoptera: Crambidae), while their agronomic performances showed no significant difference compared to the non-transgenic recipient rice. Therefore, the transgenic rice may be utilized for rice pest control in China.
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Project description:In recent years, genetically modified technology has developed rapidly, and the potential impact of genetically modified foods on human health and the ecological environment has received increasing attention. The currently used methods for testing genetically modified foods are cumbersome, time-consuming, and expensive. This paper proposed a more efficient and convenient detection method. Near-infrared diffuse reflectance spectroscopy (NIRDRS) combined with multivariate calibration methods, including principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA), and support vector machines (SVM), were used for identification of different rice varieties and transgenic (Bt63)/non-transgenic rice. Spectral pretreatment methods, including Norris-Williams smooth (NWS), standard normal variate (SNV), multiplicative scatter correction (MSC), and Savitzky-Golay 1st derivative (SG 1st-Der), were used for spectral noise reduction and effective information enhancement. Accuracy was used to evaluate the qualitative discriminant models. The results showed that the SG 1st-Der pretreatment method, combined with the SVM, provided the optimal model to distinguish different rice varieties. The accuracy of the optimal model was 98.33%. For the discrimination model of transgenic/non-transgenic rice, the SNV-SVM model, MSC-SVM model, and SG 1st-Der-PLS-DA model all achieved good analysis results with the accuracy of 100%. The results showed that portable NIR spectroscopy combined with chemometrics methods could be used to identify rice varieties and transgenic characteristics (Bt63) due to its fast, non-destructive, and accurate advantages.
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Project description:BackgroundBacillum thuringiensis (Bt) toxin produced in Cry1-expressing genetically modified rice (Bt rice) is highly effective to control lepidopteran pests, which reduces the needs for synthetic insecticides. Non-target organisms can be exposed to Bt toxins through direct feeding or trophic interactions in the field. The wolf spider Pardosa pseudoannulata, one of the dominant predators in South China, plays a crucial role in the rice agroecosystem. In this study, we investigated transcriptome responses of the 5th instar spiders fed on preys maintained on Bt- and non-Bt rice.ResultsComparative transcriptome analysis resulted in 136 differentially expressed genes (DEGs) between spiderlings preying upon N. lugens fed on Bt- and non-Bt rice (Bt- and non-Bt spiderlings). Functional analysis indicated a potential impact of Bt toxin on the formation of new cuticles during molting. GO and KEGG enrichment analyses suggested that GO terms associated with chitin or cuticle, including "chitin binding", "chitin metabolic process", "chitin synthase activity", "cuticle chitin biosynthetic process", "cuticle hydrocarbon biosynthetic process", and "structural constituent of cuticle", and an array of amino acid metabolic pathways, including "alanine, asparatate and glutamate metabolism", "glycine, serine and theronine metabolism", "cysteine and methionine metabolism", "tyrosine metabolism", "phenylalanine metabolism and phenylalanine", and "tyrosine and tryptophan biosynthesis" were significantly influenced in response to Cry1Ab.ConclusionsThe Cry1Ab may have a negative impact on the formation of new cuticles during molting, which is contributed to the delayed development of spiderlings. To validate these transcriptomic responses, further examination at the translational level will be warranted.
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Project description:Considering the anticipated commercial exploitation of insect-resistant transgenic rice and that the planting area of cultivated rice overlaps with wild rice, simulating an escape of transgenic rice from farmlands and exploring its fitness after entering semi-natural or natural ecosystems through uncontrolled seed dispersal or gene flow are critical to understand the resulting potential long-term environmental risks. The expression of foreign Cry1Ab/c protein and vegetative and reproductive fitness of insect-resistant transgenic rice Huahui1 (HH1) and its parental-line Minghui63 (MH63) were studied under four treatments combining land use and weed competition: farmland and uncultivated land under weed control (F-NW and U-NW, respectively), and farmland and uncultivated land without weed control (F-W and U-W, respectively). The expression of Cry1Ab/c was significantly lower in U-NW, F-W, and U-W than that in the control treatment, F-NW. Except for plant height, key vegetative (tiller number and biomass) and reproductive (grain number and grain weight per plant) growth indices of transgenic HH1 were significantly lower than those of the parental-line MH63 in F-NW and U-NW, indicating a significant fitness cost. In F-W and U-W, vegetative growth indices (plant height, tiller number, and biomass) were similar in HH1 and MH63; however, key reproductive indices including seed-set rate were significantly higher in HH1 than in MH63, indicating significant fitness benefits. Although these results support large-scale cultivation of insect-resistant transgenic rice in China, the ecological risk involved is high in farmland or uncultivated land without weed control (F-W and U-W).
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Project description:The environmental release and biosafety of transgenic Bt crops have attracted global attention. China has a large area of saline-alkali land, which is ideal for large-scale production of Bt transgenic rice. Therefore an understanding of the fitness of Bt transgenic rice in saline-alkaline soils and the ability to predict its long-term environmental effects are important for the future sustainable use of these crops. In the present study, we aimed to evaluate the fitness of cry1Ab/c transgenic rice in both farmland and natural ecosystems. Transgenic cry1Ab/c rice Huahui1, for which a national biosafety certificate was obtained, was grown on normal farmland and saline-alkaline soils in a glass greenhouse. The expression pattern of exogenous Cry1Ab/c protein, and vegetative and reproductive fitness of rice were assessed. The expression of the exogenous Cry1Ab/c protein in the transgenic rice grown on saline-alkaline soil was lower than that in the strain grown on farmland soil. Under both the soil conditions, vegetative growth abilities, as evaluated by tiller number and biomass, and reproductive growth abilities, as measured by filled grain number and filled grain weight per plant, showed a significantly higher fitness cost for Huahui1 than that for the parental rice Minghui63 grown under the same soil conditions. In saline-alkaline soil, the fitness cost of Huahui1was moderately higher than that of Minghui63. Therefore, the ecological risk of cry1Ab/c transgenic rice is not expected to be higher than that of parental rice Minghui63 if the former escapes into natural saline-alkaline soil. The results of the present study provide a scientific basis to improve environmental safety assessment of the insect-resistant transgenic rice strain Huahui1 before commercialization.
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Project description:BackgroundThe commercial release of rice genetically engineered to express a Cry1Ab protein from Bacillus thuringiensis (Bt) for control of Lepidoptera in China is a subject of debate. One major point of the debate has focused on the ecological safety of Bt rice on nontarget organisms, especially predators and parasitoids that help control populations of insect pests.Methodology/principal findingsA tritrophic bioassay was conducted to evaluate the potential impact of Cry1Ab-expressing rice on fitness parameters of a predaceous ground spider (Pardosa pseudoannulata (Bösenberg et Strand)) that had fed on Bt rice-fed brown planthopper (Nilaparvata lugens (Stål)) nymphs. Survival, development time and fecundity of this spider were not different when they were fed with Bt rice-fed or non-Bt rice-fed prey. Furthermore, ELISA and PCR gut assays, as well as a functional response trial, indicated that predation by P. pseudoannulata was not significantly different in Bt rice or non-Bt rice fields.Conclusions/significanceThe transgenic Cry1Ab rice lines tested in this study had no adverse effects on the survival, developmental time and fecundity of P. pseudoannulata in the laboratory or on predation under field conditions. This suggests that this important predator would not be harmed if transgenic Cry1Ab rice were commercialized.
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