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作 者:张淑红[1] 张运峰[1] 武秋颖[1] 范永山[1] ZHANG Shu-hong;ZHANG Yun-feng;WU Qiu-ying;FAN Yong-shan(Department of Life Sciences,Tangshan Normal University/Tangshan Key Laboratory of Agricultural Pathogenic Fungi and Toxins,Tangshan 063000,China)
机构地区:[1]唐山师范学院生命科学系/唐山市农业病原真菌与毒素重点实验室,河北唐山063000
出 处:《玉米科学》2023年第3期67-73,共7页Journal of Maize Sciences
基 金:唐山市人才资助项目(A201903018);唐山师范学院科研基金项目(2021B30,2021B34)。
摘 要:利用RT-PCR技术克隆玉米大斑病菌(Setosphaeria turcica)的GAPDH基因StGAPDH。该基因c DNA全长1014 bp,编码337个氨基酸;蛋白质分子量36505.44,理论pI值6.72,属弱酸、亲水、稳定蛋白,无跨膜结构域和信号肽。亚细胞定位于细胞质、线粒体和细胞核的概率分别为43.5%、30.4%和21.7%;氨基酸序列高度保守,在亲缘关系上与玉米小斑病菌(Bipolaris maydis)和水稻胡麻斑病菌(Bipolaris oryzae)最近。荧光定量PCR分析表明,在病菌接种抗病玉米自交系3 d时,StGAPDH基因表达量显著高于接种5~10 d(P<0.05);接种感病玉米自交系时,接种10 d的StGAPDH基因表达量显著高于接种3~7 d(P<0.05)。The GAPDH gene in Setosphaeria turcica(StGAPDH)was cloned by using RT-PCR technology.The cDNA length of the gene is 1014 bp,encoding 337 amino acids.The StGAPDH protein’s molecular weight is 36505.44 and the theory value of pI is 6.72,belonging to weak acid,hydrophobic,stable protein without membrane structure domain and signal peptide.The probability of subcellular localization in the cytoplasm,nucleus and mito⁃chondria is 43.5%,30.4%and 21.7%,respectively.Its amino acid sequence is highly conserved,and is phylogeneti⁃cally closed to the pathogen of Bipolaris maydis and Bipolaris oryzae.Fluorescence quantitative PCR analysis showed that the StGAPDH gene expression was significantly higher in 3 days past inoculation(dpi)than 5-7 dpi on resistant maize inbred lines(P<0.05).However,when inoculated on susceptible maize inbred lines,the gene expres⁃sion was significantly higher in 10 dpi than 3-7 dpi(P<0.05).
关 键 词:玉米 甘油醛-3-磷酸脱氢酶 玉米大斑病菌 基因克隆 序列分析 表达分析
分 类 号:S513.035.3[农业科学—作物学]
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