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作 者 ：叶雪凌,刘志勇,周宝利*

期 刊 ：西北植物学报 2013年 8期 页码：1521~1527

关键词：野生茄托鲁巴姆；黄萎病；抑制差减杂交文库；

Keywords:Solanum torvum, Verticillium wilt, suppression subtractive hybridization library,

摘 要 ：由大丽轮枝菌(Verticillium dahliae Kleb.)引起的茄子黄萎病是一种严重的土传病害,但在茄子栽培品种中缺少高抗种质,野生茄‘托鲁巴姆’高抗黄萎病。为探讨野生茄‘托鲁巴姆’抗黄萎病的分子机制,该试验以‘托鲁巴姆’为试材,利用抑制性差减杂交技术,分别以黄萎病菌侵染与未侵染的根系为检测方(tester)和驱动方(driver),构建了1个黄萎病菌诱导下的根器官正向差减cDNA文库。结果表明：（1）随机挑取170个阳性克隆,经测序,共获得109个非冗余EST,其中61个EST与其他植物的抗逆相关基因同源,如几丁质酶、细胞色素P450、过氧化物酶、蛋白酶抑制子等。（2）借助NCBI中的EST序列,经过电子拼接结合RT-PCR验证,克隆了1个通用胁迫蛋白基因——StUSP1。（3）表达分析结果显示,‘托鲁巴姆’受黄萎病菌侵染后,StUSP1在根中上调表达。

Abstract:Verticillium wilt caused by the fungus Verticillium dahliae Kleb.is a serious soilborne disease in eggplant.The eggplant cultivars are susceptible,but wild Solanum torvum is highly resistant to Verticillium wilt.A forward suppression subtractive hybridization library was established using roots of S.torvum,and the cDNAs from the fungus inoculated seedlings were used as the tester and the normal uninoculated seedlings as the driver.(1)170 positive clones were randomly selected and sequenced,and 109 non-redundant ESTs were obtained,among which 61 ESTs share sequences homology with stress resistance related genes of other plants.Such as chitinase,cytochrome P450,peroxidase,protease inhibitor Ⅱ,etc.Moreover,a EST in SSH was used as a querying probe to blast the EST database in GenBank.(2)Based on the assembled homologous cDNA sequences and RT-PCR,a universal stress gene was cloned,designated StUSP1.(3)Expression pattern revealed that StUSP1 was up-regulated in roots while S.torvum was infected by Verticillium wilt.This SSH library is helpful to understand the resistance mechanism of wild eggplant to Verticillium wilt.

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