D sR ed-M it是一种特异性定位于线粒体的荧光分子探针。本文重点探讨了D sR ed-M it探针在观察细胞凋亡形态学、以及凋亡过程中动态分子调控过程研究中的应用。实验结果表明,在细胞凋亡过程中应用该探针标记线粒体,能够直观地监测线粒...D sR ed-M it是一种特异性定位于线粒体的荧光分子探针。本文重点探讨了D sR ed-M it探针在观察细胞凋亡形态学、以及凋亡过程中动态分子调控过程研究中的应用。实验结果表明,在细胞凋亡过程中应用该探针标记线粒体,能够直观地监测线粒体肿胀的形态变化,以及细胞凋亡的重要蛋白--Bax蛋白转位线粒体和细胞色素C释放的动态过程。展开更多
Genes encoding reporter proteins are used as visual marker-assisted tools in genetic transformation as well as plant breeding. In this study, the red fluorescent protein identified in Discosoma sp. coral(DsRed2) was s...Genes encoding reporter proteins are used as visual marker-assisted tools in genetic transformation as well as plant breeding. In this study, the red fluorescent protein identified in Discosoma sp. coral(DsRed2) was successfully used as a visual marker for cotton genetic engineering. DsRed2 was successfully expressed in two cotton cultivars,JIN668 and YZ1, driven by the Ca MV-35 S promoter via the Agrobacterium-mediated transformation. Our results suggest that DsRed2 expression provides an early-stage selection tool for the transgenic calli via visual observation. Red fluorescence can be detected not only in callus and somatic embryos but also in most tissues and organs of mature plants. The transgenic line Yz-2-DsRed2 was crossed with four different cotton cultivars to assess the transgene heritability and stability in different genetic backgrounds.The heritability of the red color was highly stable when Yz-2-DsRed2 was used as a male parent. The DsRed2 gene expressed 100% in the F_1 hybrids. To investigate the relationship between DsRed2 transcription and DNA methylation, a methylation-specific PCR approach was applied to the Ca MV-35 S promoter region. The results showed a negative association between DNA methylation level in the promoter region and the transgene transcription.Taken together, these findings suggest DsRed2 a visual reporter gene for cotton genetic transformation and molecular breeding programs.展开更多
Fusarium verticillioides was labeled with DsRed via Agrobacterium tumefaciens-mediated transformation to examine differences in colonization and reactions of resistant and susceptible inbred lines of maize(Zea mays L....Fusarium verticillioides was labeled with DsRed via Agrobacterium tumefaciens-mediated transformation to examine differences in colonization and reactions of resistant and susceptible inbred lines of maize(Zea mays L.). The extent of systemic colonization of F. verticillioides in roots from maize lines either resistant or susceptible to the fungus was studied by visualizing the red fluorescence produced by the fungus expressing DsRed. The difference in quantities of colony forming units(CFU) in roots and basal stems, production of fumonisin B1, and pH of root were determined. Although F. verticillioides colonized both resistant and susceptible lines, differences were observed in the pattern and extent of fungal colonization in the two types of maize lines. The fungus colonized the susceptible lines producing mosaic patterns by filling the individual root cells with hyphae. Such a pattern of colonization was rarely observed in resistant lines, which were less colonized by the fungus than the susceptible lines in terms of CFUs. The production of mycotoxin fumonisin B1 in roots from different lines was closely correlated with the amount of F. verticillioides colonization, rather than the pH or amylopectin concentrations in the root. The findings from this study contribute to a better understanding of the defense mechanism in resistant maize lines to F. verticillioides.展开更多
文摘D sR ed-M it是一种特异性定位于线粒体的荧光分子探针。本文重点探讨了D sR ed-M it探针在观察细胞凋亡形态学、以及凋亡过程中动态分子调控过程研究中的应用。实验结果表明,在细胞凋亡过程中应用该探针标记线粒体,能够直观地监测线粒体肿胀的形态变化,以及细胞凋亡的重要蛋白--Bax蛋白转位线粒体和细胞色素C释放的动态过程。
基金supported by National Key Research and Development Program(2016YFD0100203-9)National R&D Project of Transgenic Crops of Ministry of Science and Technology of China(2016ZX08010001-006)+1 种基金Program of Introducing Talents of Discipline to Universities in China(B14032)Fundamental Research Funds for the Central Universities(2013PY064,2662015PY028,2662015PY091)
文摘Genes encoding reporter proteins are used as visual marker-assisted tools in genetic transformation as well as plant breeding. In this study, the red fluorescent protein identified in Discosoma sp. coral(DsRed2) was successfully used as a visual marker for cotton genetic engineering. DsRed2 was successfully expressed in two cotton cultivars,JIN668 and YZ1, driven by the Ca MV-35 S promoter via the Agrobacterium-mediated transformation. Our results suggest that DsRed2 expression provides an early-stage selection tool for the transgenic calli via visual observation. Red fluorescence can be detected not only in callus and somatic embryos but also in most tissues and organs of mature plants. The transgenic line Yz-2-DsRed2 was crossed with four different cotton cultivars to assess the transgene heritability and stability in different genetic backgrounds.The heritability of the red color was highly stable when Yz-2-DsRed2 was used as a male parent. The DsRed2 gene expressed 100% in the F_1 hybrids. To investigate the relationship between DsRed2 transcription and DNA methylation, a methylation-specific PCR approach was applied to the Ca MV-35 S promoter region. The results showed a negative association between DNA methylation level in the promoter region and the transgene transcription.Taken together, these findings suggest DsRed2 a visual reporter gene for cotton genetic transformation and molecular breeding programs.
基金the National Natural Science Foundation (31170080)China Agricultural Research Service (CARS-02) was greatly appreciated
文摘Fusarium verticillioides was labeled with DsRed via Agrobacterium tumefaciens-mediated transformation to examine differences in colonization and reactions of resistant and susceptible inbred lines of maize(Zea mays L.). The extent of systemic colonization of F. verticillioides in roots from maize lines either resistant or susceptible to the fungus was studied by visualizing the red fluorescence produced by the fungus expressing DsRed. The difference in quantities of colony forming units(CFU) in roots and basal stems, production of fumonisin B1, and pH of root were determined. Although F. verticillioides colonized both resistant and susceptible lines, differences were observed in the pattern and extent of fungal colonization in the two types of maize lines. The fungus colonized the susceptible lines producing mosaic patterns by filling the individual root cells with hyphae. Such a pattern of colonization was rarely observed in resistant lines, which were less colonized by the fungus than the susceptible lines in terms of CFUs. The production of mycotoxin fumonisin B1 in roots from different lines was closely correlated with the amount of F. verticillioides colonization, rather than the pH or amylopectin concentrations in the root. The findings from this study contribute to a better understanding of the defense mechanism in resistant maize lines to F. verticillioides.
