大丽轮枝菌毒素诱导表达陆地棉cDNA序列的克隆与定位

 为获得陆地棉黄萎病抗性相关候选基因，利用棉花黄萎病菌大丽轮枝菌V991毒素粗提物诱导耐黄萎病陆地棉品种'中棉12'应激基因的表达。在诱导幼根24 h时间点提取总RNA，PCR法合成双链cDNA，应用抑制消减杂交技术1]，得到了差异表达的180个克隆。经反Northern blot筛选出15个受毒素粗提物诱导应激表达的阳性cDNA克隆，将其命名为Vdrg（response gene induced by toxin of Verticillium dahliae）。本研究对所得阳性cDNA克隆进行了序列分析，并利用Pima S6 与 Acala Maxxa 海陆杂交四倍体棉花F2群体对Vdrg序列进行了棉花基因组定位。

Cloning and Mapping of cDNA from Cotton Induced by Toxin of Verticillium dahliae

In order to obtain the resistant-related candidate genes from upland cotton，response genes were induced by toxin of V. dahliae in upland cotton which display superior wilt tolerance. Total RNA was isolated from the seedling roots of upland cotton (Gossypium hirsutum L.var. Zhongmian12) after 24 h of induction by toxin of the pathogen V. dahliae strain V991, double strand cDNA was synthesized by PCR according to the procedure of the SmartTM cDNA Synthesis Kit(CLONTECH Co.), 180 clones were abtained by the method of Suppression Subtractive Hybridization,15 cDNA clones were identified to be positive by Reverse Northern Blot, which was named as response gene induced by toxin of Verticillium dahliae, for short as Vdrg. Sequence analysis and cotton genome locating of Vdrg using F2 populations derived from Pima S6(G. barbardens) ×Acala Maxxa (G. hirsutum) were also carried out in this studies.