野生茄托鲁巴姆LEA蛋白基因的克隆与序列分析

doi:10.3969/j.issn.1000-7091.2013.03.006

摘要/Abstract

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291bp,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。

关键词: 野生茄托鲁巴姆, 黄萎病, 胚胎发育晚期丰富蛋白

Abstract: 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．

Key words: Solanum torvum, Verticillium wilt, Late embriogenesis abundant protein

中图分类号:

S641

叶雪凌, 周宝利. 野生茄托鲁巴姆LEA蛋白基因的克隆与序列分析[J]. 华北农学报, 2013, 28(3): 30-34. doi: 10.3969/j.issn.1000-7091.2013.03.006.

Ye Xueling, Zhou Baoli. Cloning and Sequence Analysis of a Late Embryogenesis Abundant Protein Gene in Solanum torvum[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2013, 28(3): 30-34. doi: 10.3969/j.issn.1000-7091.2013.03.006.

http://www.hbnxb.net/CN/Y2013/V28/I3/30

参考文献

[1] Hashimoto K．Studies on Verticillium wilt of Eggplant[J]．Bulletin of the Saitama Horticultural，1989，2(3):110．
[2] Zeng H L，Ye P S，He L． Resistance evaluation of eggplant resources to Verticillium wilt in sichuan province[J]．Agricultural Science ＆ Technology，2009，10(5):123-125， 146．
[3] 连勇，刘富中，冯东昕．应用原生质体融合技术获得茄子种间体细胞杂种[J]．园艺学报， 2004， 31(1):39-42．
[4] 肖蕴华，林柏青．茄子种质资源黄萎病抗性鉴定[J]．中国蔬菜， 1995(1):32-33．
[5] 井立军，常彩涛，孙振久．茄子黄萎病抗性的杂种优势及遗传[J]．华北农学报， 2001， 16(2):58-61．
[6] 史仁玖，殷玥，王忠．野生茄子( Solanum torvum) 抗黄萎病相关基因StoVe1 的克隆与分析[J]．植物生理学通讯， 2006， 42(4):638-642．
[7] 周宝利，尹玉玲，李云鹏，等．嫁接茄根系分泌物与抗黄萎病的关系及其组分分析[J]．生态学报，2010，30(11):3073-3079．
[8] 张映霞，杨郁文，倪万潮．陆地棉黄萎病菌诱导抑制消减杂交cDNA 文库的构建与分析[J]．江苏农业学报，2008， 24(1):17-21．
[9] 朱龙付，涂礼莉，张献龙．黄萎病菌诱导的海岛棉抗病反应的SSH 文库构建及分析[J]．遗传学报，2005，32(5):528-532
[10] Soria-Guerra R E，Rosales-Mendoza S，Chang S， et al．Transcriptome analysis of resistant and susceptible genotypes of Glycine tomentella during Phakopsora pachyrhizi infection reveals novel rust resistance genes[J]． Theoretical and Applied Genetics，2010，120 (7):1315-1333．
[11] DureIII L，Crouch M，Harada J，et al． Common amino acid sequence domains among the LEA proteins of higher plants[J]． Plant Molecular Biology，1989，12: 475-486．
[12] Ingram J，Bartels D． The molecular basis of dehydration tolerance in plants[J]． Plant Molecular Biology，1996，47: 377-403．
[13] Liu R L，Liu M Q，Liu J， et al． Heterologous expression of an Ammopiptanthus mongolicus late embryogenesis abundant protein gene ( AmLEA) enhances Escherichia coli viability under cold and heat stress [J]． Plant Growth Regulation， 2010， 60(2):163- 168．
[14] 徐理，朱龙付，张献龙．棉花抗黄萎病机制研究进展[J]．作物学报， 2012， 38(9):1553-1560．
[15] Battaglia M，Olvera-Carrillo Y，Garciarrubio A． The enigmatic LEA proteins and other hydrophilins[J]． Plant Physiology， 2008， 148(1):6-24．
[16] Wise M J． LEAping to conclusions: a computational reanalysis of late embryogenesis abundant proteins and their possible roles[J]． BMC Bioinformatics， 2003，4: 52．
[17] 付桂，范伟，畅文军．海马齿胚胎发育晚期丰富蛋白基因SpLea5 的克隆与表达分析[J]．热带作物学报， 2011， 32(1):89-93．
[18] Grelet J，Benamar A，Teyssier E， et al． Identification in pea seed mitochondria of a late-embryogenesis abundant protein able to protect enzymes from drying[J]． Plant Physiology， 2005， 137:157-167．
[19] Liu Y，Wang L，Xing X， et al． ZmLEA3，a multifunctional group 3 LEA protein from maize ( Zea mays L.) ， is involved in biotic and abiotic stresses[J]． Plant Cell Physiology，2013( published online) ．

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