[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
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.
