中国农业科学 ›› 2010, Vol. 43 ›› Issue (10): 1977-1984 .doi: 10.3864/j.issn.0578-1752.2010.10.001

• 作物遗传育种·种质资源·分子遗传学 •    下一篇

条锈菌诱导的小麦TaHin1的克隆与表达特征分析

邓麟,王晓杰,刘新颖,蔡高磊,汤春蕾,魏国荣,黄丽丽,康振生

  

  1. (西北农林科技大学植物保护学院)

  • 收稿日期:2009-12-02 修回日期:2009-12-29 出版日期:2010-05-15 发布日期:2010-05-15
  • 通讯作者: 康振生

Isolation and Expression Analysis of a TaHin1 Gene Induced by Stripe Rust Fungus in Wheat

DENG Lin,WANG Xiao-jie, LIU Xin-ying, CAI Gao-lei, TANG Chun-lei, WEI Guo-ron1, HUANG Li-li, KANG Zhen-sheng
  

  1. (西北农林科技大学植物保护学院)

  • Received:2009-12-02 Revised:2009-12-29 Online:2010-05-15 Published:2010-05-15
  • Contact: KANG Zhen-sheng

摘要:

【目的】克隆受条锈菌诱导的小麦Hin1,研究其在小麦抗条锈病防御反应以及抗非生物胁迫过程中的作用。【方法】通过电子克隆、RT-PCR方法,从条锈菌侵染的小麦水源11中分离出一个编码HIN1的cDNA序列;通过生物信息学工具对DNA序列及其编码的蛋白序列进行分析;利用实时荧光定量PCR分析该基因在小麦不同器官、与条锈菌互作、外源激素处理以及非生物胁迫下的表达情况。【结果】分离得到小麦Hin1,命名为TaHin1,开放阅读框为642 bp,编码213个氨基酸,分子量为23.24 kD,等电点8.67,具有信号肽和HIN1结构域,可能为分泌蛋白;与高粱和水稻HIN1同源性达80%;表达分析结果表明,TaHin1在小麦根、茎、叶组织中表达差异不显著,在小麦与条锈菌互作时,仅在非亲和组合中被诱导表达;外源植物激素水杨酸、茉莉酸诱导TaHin1上调表达,脱落酸诱导其下调表达,而乙烯不诱导其表达;高盐、干旱、机械损伤以及低温等非生物胁迫下,TaHin1均上调表达。【结论】首次克隆到一个受条锈菌诱导的小麦TaHin1,可能通过水杨酸、茉莉酸信号途径参与了小麦对条锈菌的防御反应,而脱落酸在防御反应中起负调控作用,该基因在小麦对非生物胁迫的抗逆应答过程中也发挥着一定作用。

关键词: 条锈菌, TaHin1, 电子克隆, 表达特征

Abstract:

【Objective】 A novel Hin1 gene induced by the stripe rust pathogen was cloned from wheat to better understand its role in wheat defense to Puccinia striiformis f.sp. tritici and to abiotic stresses. 【Method】 In-silico cloning and RT-PCR were combined to isolate a gene which might encode HIN1 from wheat Suwon 11 infected by stripe rust pathogen. Bioinformatics tools were applied to analyze both the DNA sequence and protein sequence. The expression patterns of the gene in wheat organs, as well as interactions with stripe rust pathogen, induction of exogenous phytohormones and abiotic stresses, were investigated using real time quantitative RT-PCR. 【Result】 The wheat Hin1 gene designated as TaHin1 was obtained. Open reading frame (ORF) of TaHin1 was 642 bp in length, encoding 213 amino acid residues with a molecular weight of 23.24 kD and predicted pI value of 8.67. TaHIN1 might be a secreted protein, containing a signal peptide and a HIN1 domain. TaHIN1 shared about 80% similarity with HIN1 from broomcorn and rice. TaHin1 expresses in wheat leaf, stem and root with no significant difference. Challenged by stripe rust fungus, TaHin1 was induced by Puccinia striiformis f.sp. tritici only in incompatible interaction. TaHin1 was up-regualted by exogenous salicylic acid and jasmonic acid while down-regulated by abascisci acid. Exogenous ethylene could not induce the expression. TaHin1 was obviously up-regulated by various abiotic stresses such as high salinity, drought, mechanical wound and low temperature. 【Conclusion】 TaHin1 gene was firstly cloned and characterized from wheat infected by stripe rust fungus. TaHin1 might facilitate wheat defence through salicylic acid and jasmonic acid pathways, while abscisci acid might play a negative role in the defence. TaHin1 also played a role in response to abiotic stresses.

Key words: stripe rust, TaHin1, in-silico cloning, transcriptional profiling