作物学报 ACTA AGRONOMICA SINICA 2010, 36(6): 953−960 http://www.chinacrops.org/zwxb/ ISSN 0496-3490; CODEN TSHPA9 E-mail: xbzw@chinajournal.net.cn 本 究 国家 科 基金 点 (30930064), 代 产 技 建 金和高等 科创 计划 (B07049) 。 * (Corresponding author): 康 , E-mail: kangzs@nwsuaf.edu.cn 第 联 方 : E-mail: liuxy3600902@163.com ** 共 第 Received( 稿 ): 2010-01-08; Accepted(接 ): 2010-03-04. DOI: 10.3724/SP.J.1006.2010.00953 小麦钙调素新亚型 TaCaM5 的克隆及表达分析 刘新颖 1 王晓杰 1,** 薛 杰 1 夏 宁 1 邓 麟 1 蔡高磊 1 汤春蕾 1 魏国荣 1 黄丽丽 1 康振生 1,2,* 1 北 科技大 保护 , 712100; 2 北 科技大 / 分 点 , 712100 : 利 RT-PCR 技 , 从 菌 导的 分离出 个编 CaM 基 的 cDNA 列, 经氨基 列分 定 个 的 CaM , 被 TaCaM5。TaCaM5 包含 个 450 bp 的开放 读框, 编 149 个氨基 ; 编 的蛋白不含跨 、 号 、定 胞 , 具 4 个 EF-hand 保 结构 。 的 CaM 基 , TaCaM5 CaM 基 的 关 近, 高达 97%。该基 根、茎、 等 均 不 程度的表 达; 并 菌 导表达, 非 和 合 和 合 , 分别 接 后 6 h 和 24 h 表达量 高。 激 、 莉 甲 和 导 TaCaM5 调表达, 导 调表达。TaCaM5 机 害、干旱和低 件 表 达量 , 高 环境 表达量降低。表 TaCaM5 可 过 莉 和 等 号 径参 对 菌的防 反 , 参 机 害、低 和干旱环境 的 Ca 2+ -CaM 号 导 径。 关键词: 病; 钙调 ; 非 ; 光定量 RT-PCR Cloning and Expression Analysis of a Novel Calmodulin Isoform TaCaM5 from Wheat LIU Xin-Ying 1 , WANG Xiao-Jie 1,** , XIA Ning 1 , DENG Lin 1 , CAI Gao-Lei 1 , TANG Chun-Lei 1 , WEI Guo-Rong 1 , HUANG Li-Li 1 , and KANG Zhen-Sheng 1,2,* 1 College of Plant Protection, Northwest A&F University, Yangling 712100, China; 2 Shaanxi Provincial Key Laboratory of Molecular Biology for Agriculture / Northwest A&F University, Yangling 712100, China Abstract: Calmodulin is a ubiquitous transducer of calcium signals in eukaryotes. It mediates a lot of cellular processes, such as transcription, cytoskeletal organization and motility, and amino acid metabolism. In diploid plant species, several isoforms of calmodulin have been described. A novel CaM gene was isolated from cDNA library of wheat leaves infected by Puccinia strii- formis f. sp. tritici through RT-PCR approach. The gene was tentatively designated as TaCaM5 and encoded a novel CaM isoform based on protein sequence analysis. This open reading frame of TaCaM5 was 450 bp in length and 149 amino acids were encoded with four conserved EF-hand domains. TaCaM5, in which transmembrane domain or signal peptide sequence was absent, was predicted existing in cytoplasm. The amino acid sequence of TaCaM5 shares 97% identify with ZmCaM from Zea mays. TaCaM5 expressed differently in wheat leaf, stem, and root. Challenged by stripe rust fungus (Puccinia striiformis f. sp. tritici), TaCaM5 was induced by this fungus in both incompatible and compatible interactions, with the maximal expression at 6 h and 24 h post inoculation respectively. TaCaM5 was up-regualted by exogenous abscisic acid, ethylene, and jasmonic acid but down-regulated by salicylic acid. TaCaM5 was obviously up-regulated by various abiotic stresses, such as low temperature, mechanical wound, and drought. However, high salinity stress could not induce the expression of TaCaM5. These results suggest that TaCaM5 is probably involved in regulating the host defence responses through ethylene and jasmonic acid pathways, and also participate in Ca 2+ -CaM signal transmission pathways under mechanical wound, low temperature, and drought conditions. Keywords: Wheat stripe rust; CaM isoform; Abiotic stress; qRT-PCR 柄 菌(Puccinia sriiformis f. sp. tritici) 的 病 界范 广泛分布的 类 部病害, 大 积暴发 产量, 成绝 。 践 , 和合理利 抗病