条锈菌,实时荧光定量PCR,表达谱,TaOZR,活性氧," /> 条锈菌,实时荧光定量PCR,表达谱,TaOZR,活性氧,"/> stripe rust,qRT-PCR,expression profiling,TaOZR,activated oxygen species
,"/> <font face="Verdana">条锈菌诱导的小麦TaOZR的克隆及特征分析</font>

中国农业科学 ›› 2010, Vol. 43 ›› Issue (12): 2403-2409 .doi: 10.3864/j.issn.0578-1752.2010.12.002

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

条锈菌诱导的小麦TaOZR的克隆及特征分析

蔡高磊,王晓杰,刘丹,邓麟,刘新颖,汤春蕾,赵杰,魏国荣,黄丽丽,康振生   

  1. (西北农林科技大学植物保护学院/陕西省农业与分子生物学重点实验室)
  • 收稿日期:2010-01-07 修回日期:2010-02-07 出版日期:2010-06-15 发布日期:2010-06-15
  • 通讯作者: 康振生

Isolation and Expression Analysis of a TaOZR Gene Induced by Stripe Rust Fungus in Wheat#br#

CAI Gao-lei, WANG Xiao-jie, LIU Dan, DENG Lin, LIU Xin-ying, TANG Chun-lei, ZHAO Jie, WEI Guo-rong, HUANG Li-li, KANG Zhen-sheng#br#   

  1. (西北农林科技大学植物保护学院/陕西省农业与分子生物学重点实验室)
  • Received:2010-01-07 Revised:2010-02-07 Online:2010-06-15 Published:2010-06-15
  • Contact: KANG Zhen-sheng

摘要:

【目的】克隆受条锈菌诱导的小麦OZR,研究其在小麦抗条锈病防御反应及非生物胁迫过程中的作用。【方法】通过电子克隆及RT-PCR方法,从条锈菌侵染的小麦水源11中分离出1个编码OZR的cDNA序列。利用生物信息学对cDNA序列及其编码的蛋白序列进行分析;采用实时荧光定量PCR(qRT-PCR)分析该基因在小麦与条锈菌互作、外源激素处理以及非生物胁迫下的诱导表达情况。【结果】获得小麦OZR,命名为TaOZR。其ORF长240 bp,编码79个氨基酸;蛋白序列理论分子量8.67 kD,等电点9.34;具有信号肽和跨膜区,可能为分泌蛋白;与水稻OZR相似性达76%;TaOZR受条锈菌诱导后在亲和互作中差异不显著,在非亲和互作中诱导表达;外源植物激素水杨酸、乙烯、脱落酸、茉莉酸均诱导TaOZR的积累并上调表达;低温、干旱、高盐非生物胁迫下TaOZR表达差异不显著。【结论】首次克隆到一个条锈菌诱导的小麦TaOZR,TaOZR可能通过水杨酸、乙烯、脱落酸和茉莉酸信号途径协同参与小麦对条锈菌的早期防御。

关键词: 条锈菌')">条锈菌, 实时荧光定量PCR, 表达谱, TaOZR, 活性氧

Abstract:

【Objective】 A novel OZR gene was cloned from wheat induced by the stripe rust pathogen, and the expression levels were evaluated in wheat challenged by Puccinia striiformis f. sp. tritici or abiotic stress. 【Method】 A cDNA sequence was isolated from wheat cultivar Suwon 11 infected by stripe rust pathogen using in silico cloning and RT-PCR methods. Bioinformatics tools were applied to analyse both the cDNA sequence and protein sequence. The expression patterns of the gene in the interaction of wheat and stripe rust pathogen, exogenous phytohomones and abiotic stresses were investigated using real time quantitative (qRT-PCR). 【Result】 The wheat OZR gene was obtained from wheat infected by stripe rust pathogen, which was designated TaOZR. The opening reading frame (ORF) of TaOZR was 240 bp in length, encoding 79 amino acids with molecular weight 8.67 kD and theoretical pI 9.34. TaOZR containing a signal peptide and a transmembrane might be a secreted protein. TaOZR shared 76% similarity with rice. The transcriptional accumulation of TaOZR was increased in incompatible interaction, and unchanged in compatible interaction. The expression levels of TaOZR were up-regualted by exogenous hormone salicylic acid, ethylene, abascisci acid, jasmonic acid treatments. However, the transcripts of TaOZR had no change in response to various abiotic stresses like low temperature, drought and high salinity. 【Conclusion】 TaOZR gene was firstly cloned and characterized from wheat infected by stripe rust fungus. TaOZR might facilitate wheat defence to stripe rust pathogen through salicylic acid ethylene, abascisci acid, jasmonic acid signal pathways.

Key words: stripe rust')">stripe rust, qRT-PCR, expression profiling, TaOZR, activated oxygen species