条锈菌诱导的小麦TaOZR的克隆及特征分析.pdfVIP

中国农业科学 2010,43(12):2403-2409 Scientia Agricultura Sinica doi: 10.3864/j.issn.0578-1752.2010.12.002 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 （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，陕西杨凌 712100 ） 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；实时荧光定量PCR；表达谱；TaOZR；活性氧 Isolation and Expression Analysis of a TaOZR Gene Induced by Stripe Rust Fungus in Wheat 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 (College of Plant Protection, Northwest Agricultural and Forestry University/Shaanxi Key Laboratory of Molecular Biology for Agriculture, Yangling 712100, Shaanxi) 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