转hpa1_(Xoo)基因棉花活性氧产生及防卫相关基因的表达  

The generation of reactive oxygen intermediates and expression of defense genes in transgenic hpa1_(Xoo) cotton

作　　者：缪卫国[1] 宋从凤[2] 乔子辰[3] 王金生[2] 

机构地区：[1]海南大学环境与植物保护学院,海南海口570228 [2]南京农业大学植物保护学院/农业部作物病虫害监测与防控重点开放实验室,江苏南京210095 [3]新疆农业大学农学院,新疆乌鲁木齐830052

出　　处：《南京农业大学学报》2011年第2期61-66,共6页Journal of Nanjing Agricultural University

基　　金：国家自然科学基金项目(30230240);国家973计划项目(G2000016201);国家科技支撑项目(2004BA901A36)

摘　　要：采用H2O2和酶活性检测、DAB染色、定量实时PCR等方法研究转hpa1Xoo基因棉花T-34叶组织中的活性氧。结果显示:无病原菌侵染时,T-34和出发品种Z35均未产生活性氧。在接种黄萎病菌分生孢子悬浮液后,与Z35相比,在T-34叶组织中接菌0～8 h H2O2含量变化呈现2个峰值;接菌8 h时PAL和POD酶活性显著增强;DAB染色显示褐红色斑产生;2个防卫基因ghAOX1和hsr203J超表达。这些结果表明,转hpa1Xoo基因棉花为某些防卫基因超水平表达提供了分子基础,使受到病原菌侵染时才诱导产生的高水平活性氧及相关基因表达,并且组成型表达的HarpinXoo赋予转基因棉花对黄萎病菌的抗性。In this study,respective methods in detection of H2O2 and relative enzyme activity,DAB staining and quantitative real time PCR were used for confirming the reactive orygen intermediates（ROI）generation in T-34 compared with untransformed cotton Z35.The results showed that after T-34 leaves were inoculated by Verticillium dahliae conidia,compared with Z35 leaves,H2O2 content in T-34 leaf tissue exhibited two peak values during 0-8 h;PAL and POD activities were significantly enhanced,DAB staining revealed visual reddish-brown coloration spots,and expressions of ghAOX1 and hsr203J genes were significantly induced.These evidences indicated that HarpinXoo expressing cotton possessed the molecular basis for expression of some defense genes,and the expression of HarpinXoo was responsible for the generation of ROI and the expression of the related defense gene in transgenic hpa1Xoo plant,and conferred the resistance of transgenic hpa1Xoo cotton to Verticillium dahliae.

关 键 词：HarpinXoo 转基因棉花 活性氧 PAL POD 防卫基因 

分 类 号：S435.72[农业科学—农业昆虫与害虫防治]