论文部分内容阅读
棉花黄萎病是危害棉花维管束的严重病害,从陆地棉、海岛棉及近缘野生种杂交后代中筛选抗黄萎病种质能有效拓宽陆地棉抗源基础。本研究通过人工接种强致病力、落叶型大丽轮枝菌(Verticillium dahliae Kleb.)菌株Vd991,对包括海岛棉(G.barbadense)、陆地棉(G.hirsutum)和瑟伯氏棉(G.thurberi Todro)三元杂种衍生系在内的42份棉花种质的抗病性进行了鉴定,并以棉花Gh ACTIN14(Gen Bank:AY305733)为内标,设计抗病相关基因Gb DIR1、Gb DIR2、β-1,3葡聚糖酶基因的q RT-PCR(quantitative Real-time PCR)特异引物,使用SYBR Green PCR试剂盒标记反应产物,在Applied Biosystems 7500 Real-Time PCR System上采用2-ΔΔCt法分析高抗黄萎病的三元杂种衍生系海陆野96-5受黄萎病菌侵染的调控表达特征。结果显示:筛选出的16份抗黄萎病种质中有2份高抗种质和12份抗病种质均为陆海瑟三元杂交种衍生系或衍生品种;q RT-PCR分析表明高抗病品系海陆野96-5受黄萎病菌侵染1 h时,Gb DIR1、Gb DIR2和β-1,3葡聚糖酶即开始上调表达,Gb DIR1到8 h时上调最高,达到处理前20倍,Gb DIR2则在24 h和96 h表达量较高,分别上调15倍和29倍,β-1,3葡聚糖酶在8 h和48 h时上调表达2倍以上。本研究结果为高抗黄萎病资源的利用奠定了基础。
Verticillium dahliae is a serious disease that endangers cotton vascular bundles. Screening anti-Verticillium wilt germplasm from G. hirsutum, G. japonica and related wild relatives can effectively broaden the anti-source base of upland cotton. In this study, the virulent, Verticillium dahliae Kleb. Strain Vd991 was inoculated artificially inoculation with G. barbadense, G.hirsutum and Cypress (G .Thurberi Todro) derived from 42 cotton germplasms were identified and the resistance genes Gb DIR1 and Gb DIR2 were designed with cotton Gh ACTIN14 (Gen Bank: AY305733) Q-1,3 glucanase gene by q RT-PCR (specific Real-time PCR) specific primers, using the SYBR Green PCR kit for labeling the reaction products on an Applied Biosystems 7500 Real-Time PCR System using 2-ΔΔCt Analysis of Regulated Expression Characteristics of Rhizoctonia solani 96-5 Derived from Verticillium dahliae Derived from Vertically Derived Derivative Line Hailuye. The results showed that 2 out of 16 cross-resistance germplasms and 12 resistant germplasms were derived from Luheseer ternary hybrids or derived lines; q RT-PCR analysis showed that high resistance The Gb DIR1, Gb DIR2 and β-1, 3-glucanase began to up-regulate when infected by Verticillium dahliae at 96-5. The Gb DIR1 up-regulated at 8 h and reached the level of 20 Gb DIR2 was up-regulated by 15-fold and 29-fold at 24 h and 96 h, respectively. The expression of β-1,3-glucanase was upregulated more than 2-fold at 8 h and 48 h. The results of this study laid the foundation for the utilization of high resistance to Verticillium wilt.