野生茄LEA 蛋白基因的克隆与特征分析.doc


野生茄 LEA 蛋白基因的克隆与特征分析#
叶雪凌,刘志勇,周宝利**
(沈阳农业大学园艺学院,沈阳 110866)
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摘要:黄萎病是危害茄子生产的一种严重真菌病害。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。前期研究中,本实验室利用抑制消减杂交技术构建了野生茄“托鲁巴姆”
幼苗根器官经黄萎病菌侵染后的 cDNA 文库。在本研究中,选取其中 1 个 EST,结合电子
克隆及 RT-PCR 验证的策略,获得了胚胎发育晚期丰富蛋白基因的全长 cDNA 序列,命名
为 StLEA1。该基因的完整开放阅读框为 291bp,编码 96 个氨基酸,编码蛋白的分子量
kD, 等电点为 。经序列分析,该蛋白编码水溶性蛋白,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,当“托鲁巴姆”受黄
萎病菌侵染后,StLEA1 在根系中上调表达。
关键词:野生茄;黄萎病;胚胎发育晚期丰富蛋白
中图分类号:
Cloning and characterization of a late embryogenesis
abundant protein gene in Solanum Torvum
YE Xueling, LIU Zhiyong, Zhou Baoli
(Department of Horticulture,Shenyang Agriculture University,Shenyang 110866)
Abstract: Verticillium wilt is a serious fungal disease that affects eggplant plants. Solanum
Torvum has high resistance to Verticillium wilt. Therefore it is the ideal material for studying the
mechanism of resistance to Verticillium wilt in Solanum Torvum . Late embryogenesis abundant
proteins ( LEA proteins) are a group of stress-responsive proteins in higher plants that are induced
by environmental stress. In our precious study, analysis of gene differential expression was
performed by SSH to isolate up-regulated genes in the roots of Solanum Torvum while infected by
Verticillium wilt. A differential TDF (Transcript Derived Fragments) i