Extensive planting of Bacillus thuringiensis （Bt）-transgenic plants economically benefits society; how-ever, the potential risk they pose is receiving increasing attention. This study used enzyme-linked immunosorben...Extensive planting of Bacillus thuringiensis （Bt）-transgenic plants economically benefits society; how-ever, the potential risk they pose is receiving increasing attention. This study used enzyme-linked immunosorbent assay and fluorescence quantitative PCR （RT-PCR） to monitor the temporal and spatial dynamics of the expression of Bt toxic protein in a forest of 6- to 8-year-old trees of transgenic insect-resistant poplar 741 for three consec- utive years. The enrichment, distribution, and degradation of Bt toxic protein and the influence of transgenic poplars on the targeted insect population, Hyphantria cunea, were investigated. The content of CrylAc toxic protein dynamically changed in transgenic poplar. During the annual growth cycle, the content initially increased, then decreased in the long and the short branches of the crown and in the root system, peaking in August. During the study, the protein did not accumulate overtime. The mRNA transcription of gene CrylAc was almost consistent with the level of the protein, but transcription peaked in July. In the transgenic and control forestland, microscale levels of the CrylAc toxic protein were detected from the soil, but increased accumulation was not observed with the planting year of transgenic poplar. Meanwhile, Bt was isolated and detected molecularly from the soil in the experimental forestland. A systematic investigation of the density of H. cunea in the experimental transgenic poplar forest indi- cated that transgenic Pb29 poplar could resist insects to a certain degree. At peak occurrence of the targeted insects, the density of H. cunea in the experimental forest was significantly lower than in the nontransgenic poplar forest.展开更多
[Objective] The paper was to study the expression characteristics and borer resistance of Bt gene of Songjing 9 （1C^＊）. [Method] With super japonica rice Songjing 9 as the receptor, transgenic crylC^＊ super japon...[Objective] The paper was to study the expression characteristics and borer resistance of Bt gene of Songjing 9 （1C^＊）. [Method] With super japonica rice Songjing 9 as the receptor, transgenic crylC^＊ super japonica rice Songjing 9 （1C^＊） regulated by ubi promoter was created by agrobacterium-mediated genetic transformation method. Ten strains of transgenic super japonica rice formed from different transformation events were planted in the field; the Tt-mRNA of leaf, stem-sheath and young spikelet at booting stage were detected by real-time fluorescent quantitative PCR; the Bt protein content of leaf, stem-sheath and young spikelet and harvested brown rice were detected by enzyme-linked immunosorbent assay （ELISA）; the white head rate damaged by Chilo suppressalis was manually investigated. [Result] The Bt-mRNA and Bt protein content of various organs and Bt protein content of mature brown rice at booting stage of Songjing 9 （1C^＊） significantly differed among strains; there was no correlation relationship in Bt-mRNA and Bt protein content between leaf and stem-sheath, between leaf and young spikelet, and between stem-sheath and young spikelet; there was no relation between Bt-mRNA or Bt protein content of leaf, stem-sheath and young spikelet and Bt protein content of brown rice; there was positive correlation between Bt-mRNA and Bt protein content in the same organ such as leaf, stem-sheath and young spikelet at booting stage; such strains as Songjing 9 （1C^＊）, whether the Bt gene expression was high or low, showed good resistance against C.suppressalis. This indicated that there were differences in Bt gene expression in different strains of transgenic cry1C^＊ super japonica rice or different organs of the same strain; there was no relation in Bt gene expression among different organs; the Bt-mRNA and protein content of the same organ usually had consistent expression; the Bt gene expression of various organs had nothing to do with the Bt protein content of brown rice. [Conclusion] The Bt gene and its expression cassette can meet the requirement of cultivating japonica rice varieties against stem borers.展开更多
[Objective] The paper was to study the temporal and spatial dynamics of Bt protein expression in transgenic Bt cotton and to determine the inner relationship of Bt protein expression and transgenic Bt cotton. [Method]...[Objective] The paper was to study the temporal and spatial dynamics of Bt protein expression in transgenic Bt cotton and to determine the inner relationship of Bt protein expression and transgenic Bt cotton. [Method] With transgenic cotton cultivar（ GK19） as the test material,Bt protein contents in different organs,main stem functional leaves at different growth stages and different positions of main stem leaves at different growth stages were studied by enzyme-linked immunosorbent assay. [Result] There were differences in Bt protein content among different organs of transgenic Bt cotton; the Bt protein content of leaves at seedling stage was the highest,followed by flowers,bubs and bolls,and those of roots and stems were relatively low. The Bt protein content of main stem function leaves gradually decreased with the progressing development. There were great differences in Bt protein content among different positions of main stem leaves at different growth stages; the Bt protein content of the 1^（st）-7^（th） top leaves at seedling stage and full budding stage gradually decreased,while those at full flowering stage and full bolling stage first slowly increased then gradually stabilized. [Conclusion] Bt protein expression was found in all organs of transgenic cotton at all growth stages,and the expression level presented temporal and spatial dynamics.展开更多
基金supported by the National High Technology Research and Development Program of China(863 Program)(Grant No.2013AA102703)
文摘Extensive planting of Bacillus thuringiensis （Bt）-transgenic plants economically benefits society; how-ever, the potential risk they pose is receiving increasing attention. This study used enzyme-linked immunosorbent assay and fluorescence quantitative PCR （RT-PCR） to monitor the temporal and spatial dynamics of the expression of Bt toxic protein in a forest of 6- to 8-year-old trees of transgenic insect-resistant poplar 741 for three consec- utive years. The enrichment, distribution, and degradation of Bt toxic protein and the influence of transgenic poplars on the targeted insect population, Hyphantria cunea, were investigated. The content of CrylAc toxic protein dynamically changed in transgenic poplar. During the annual growth cycle, the content initially increased, then decreased in the long and the short branches of the crown and in the root system, peaking in August. During the study, the protein did not accumulate overtime. The mRNA transcription of gene CrylAc was almost consistent with the level of the protein, but transcription peaked in July. In the transgenic and control forestland, microscale levels of the CrylAc toxic protein were detected from the soil, but increased accumulation was not observed with the planting year of transgenic poplar. Meanwhile, Bt was isolated and detected molecularly from the soil in the experimental forestland. A systematic investigation of the density of H. cunea in the experimental transgenic poplar forest indi- cated that transgenic Pb29 poplar could resist insects to a certain degree. At peak occurrence of the targeted insects, the density of H. cunea in the experimental forest was significantly lower than in the nontransgenic poplar forest.
基金Supported by National High Technology Research and Development Program(863 plan)of China(2014AA10A600)National Major Project for Transgenic Organism Breeding of China(2016ZX08001001-001-007)Open Project of Key Laboratory of Molecular Biology in Colleges and Universities of Heilongjiang Province(1206)
文摘[Objective] The paper was to study the expression characteristics and borer resistance of Bt gene of Songjing 9 （1C^＊）. [Method] With super japonica rice Songjing 9 as the receptor, transgenic crylC^＊ super japonica rice Songjing 9 （1C^＊） regulated by ubi promoter was created by agrobacterium-mediated genetic transformation method. Ten strains of transgenic super japonica rice formed from different transformation events were planted in the field; the Tt-mRNA of leaf, stem-sheath and young spikelet at booting stage were detected by real-time fluorescent quantitative PCR; the Bt protein content of leaf, stem-sheath and young spikelet and harvested brown rice were detected by enzyme-linked immunosorbent assay （ELISA）; the white head rate damaged by Chilo suppressalis was manually investigated. [Result] The Bt-mRNA and Bt protein content of various organs and Bt protein content of mature brown rice at booting stage of Songjing 9 （1C^＊） significantly differed among strains; there was no correlation relationship in Bt-mRNA and Bt protein content between leaf and stem-sheath, between leaf and young spikelet, and between stem-sheath and young spikelet; there was no relation between Bt-mRNA or Bt protein content of leaf, stem-sheath and young spikelet and Bt protein content of brown rice; there was positive correlation between Bt-mRNA and Bt protein content in the same organ such as leaf, stem-sheath and young spikelet at booting stage; such strains as Songjing 9 （1C^＊）, whether the Bt gene expression was high or low, showed good resistance against C.suppressalis. This indicated that there were differences in Bt gene expression in different strains of transgenic cry1C^＊ super japonica rice or different organs of the same strain; there was no relation in Bt gene expression among different organs; the Bt-mRNA and protein content of the same organ usually had consistent expression; the Bt gene expression of various organs had nothing to do with the Bt protein content of brown rice. [Conclusion] The Bt gene and its expression cassette can meet the requirement of cultivating japonica rice varieties against stem borers.
基金Supported by Genetic Engineering Project of Sichuan Province(2014LWJJ-011)
文摘[Objective] The paper was to study the temporal and spatial dynamics of Bt protein expression in transgenic Bt cotton and to determine the inner relationship of Bt protein expression and transgenic Bt cotton. [Method] With transgenic cotton cultivar（ GK19） as the test material,Bt protein contents in different organs,main stem functional leaves at different growth stages and different positions of main stem leaves at different growth stages were studied by enzyme-linked immunosorbent assay. [Result] There were differences in Bt protein content among different organs of transgenic Bt cotton; the Bt protein content of leaves at seedling stage was the highest,followed by flowers,bubs and bolls,and those of roots and stems were relatively low. The Bt protein content of main stem function leaves gradually decreased with the progressing development. There were great differences in Bt protein content among different positions of main stem leaves at different growth stages; the Bt protein content of the 1^（st）-7^（th） top leaves at seedling stage and full budding stage gradually decreased,while those at full flowering stage and full bolling stage first slowly increased then gradually stabilized. [Conclusion] Bt protein expression was found in all organs of transgenic cotton at all growth stages,and the expression level presented temporal and spatial dynamics.
