野生茄托鲁巴姆LEA蛋白基因的克隆与序列分析  

Cloning and Sequence Analysis of a Late Embryogenesis Abundant Protein Gene in Solanum torvum

作　　者：叶雪凌[1] 周宝利[1] 

机构地区：[1]沈阳农业大学园艺学院,辽宁沈阳110866

出　　处：《华北农学报》2013年第3期30-34,共5页Acta Agriculturae Boreali-Sinica

基　　金：高等学校博士点专项科研基金项目(20102103120005);辽宁省博士启动基金项目(20101104);辽宁省教育厅项目(L2010494);中国博士后基金项目(20100471472)

摘　　要：野生茄托鲁巴姆高抗黄萎病,是研究茄子黄萎病抗性的理想试材。胚胎发育晚期丰富蛋白(LEA蛋白)是植物在逆境条件下生成的一类应激蛋白。研究以1个受黄萎病菌诱导的托鲁巴姆EST为种子序列,结合电子克隆及RT-PCR验证的策略,获得了LEA蛋白基因的全长cDNA序列,命名为StLEA1。该基因的完整开放阅读框为291 bp,编码96个氨基酸,编码蛋白的分子量为10.748 kDa,等电点为9.913。经序列分析,StLEA1为水溶性蛋白,不存在信号肽,具有LEA5家族的典型结构域和保守序列,预测含有多个磷酸化位点。表达分析表明,托鲁巴姆受黄萎病菌侵染后,StLEA1在根系中上调表达。为探讨野生茄托鲁巴姆抗黄萎病分子机制提供了素材。Solanum torvum,a wild species of eggplant,is highly resistant to Verticillium wilt.Therefore,it is the ideal material for studying the mechanism of resistance to Verticillium wilt in eggplant.Late embryogenesis abundant proteins（LEA proteins） are a group of stress-responsive proteins in higher plants that are induced by environmental stress.In this study,a up-regulated EST while infected by Verticillium wilt was used as a querying probe to blast the GenBank database.Based on the assembled homologous cDNA sequences,a 490 bp cDNA was amplified and cloned by RT-PCR,designated StLEA1.The ORF of StLEA1 is 291 bp,coding a protein with 96 animo acids.The molecular weight of coding protein is 10.748 kDa and isoelectric point is 9.913.StLEA1 protein is a soluble protein with multiple phosphorylation sites,conserve domain in Lea5,but without signal peptides.Quantitative RT-PCR analysis revealed that StLEA1 was up-regulated in roots while Solanum torvum was infected by Verticillium wilt.The result provides the material for studying the resistance mechanism of wild eggplant to Verticillium wilt.

关 键 词：野生茄托鲁巴姆 黄萎病 胚胎发育晚期丰富蛋白 

分 类 号：S641[农业科学—蔬菜学]