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从小麦中克隆和鉴定钙结合 EF 手蛋白基因 TaCab1 及其对条锈菌和非生物胁迫的表达。

Cloning and characterization of a calcium binding EF-hand protein gene TaCab1 from wheat and its expression in response to Puccinia striiformis f. sp. tritici and abiotic stresses.

机构信息

College of Plant Protection and Shaanxi Key Laboratory of Molecular Biology for Agriculture, Northwest A&F University, Yangling, 712100, Shaanxi, People's Republic of China.

出版信息

Mol Biol Rep. 2011 Aug;38(6):3857-66. doi: 10.1007/s11033-010-0501-8. Epub 2010 Nov 26.

Abstract

Calcium is a ubiquitous and essential secondary messenger in eukaryotic signal transduction pathways. Calcium binding protein, as a component of pathways, plays various roles in response to biotic and abiotic stresses, as well as in developmental processes in plants. In this study, a calcium binding protein gene, designated as TaCab1 (Triticum aestivum calcium binding EF-hand protein 1), was isolated and characterized from wheat leaves (cv. Suwon 11) infected by Puccinia striiformis f. sp. tritici by in silico cloning and reverse transcription PCR (RT-PCR). TaCab1 did not have an intron and was predicted to encode a 216 amino acid protein which possesses an N-terminal region with a signal peptide, a transmembrane domain, an EF-hand motif and a caleosin domain. The results of transient assays with constructs of TaCab1 with green fluorescent protein (GFP) gene indicated that TaCab1 encodes a transmembrane protein. Quantitative real-time PCR (qRT-PCR) analyses revealed that TaCab1 was highly expressed in leaves than roots and stems. Although up-regulated expression profiles of TaCab1 were quite similar in both incompatible and compatible interactions, its transcript accumulation in the compatible interaction was much higher than in the incompatible interaction. The transcription of TaCab1 was also up-regulated at different degrees after treated by phytohormones [abscisic acid, benzyl adenine, ethylene, methyl jasmonate and salicylic acid (SA)] and stress stimuli [wounding, low temperature, polyethylene glycol and high salinity]. These results suggest that TaCab1 is involved in the plant-pathogen recognition, symptom development, and the basal tolerance to biotic and abiotic stresses through the SA signaling pathway.

摘要

钙是真核信号转导途径中普遍存在且必不可少的二级信使。钙结合蛋白作为途径的组成部分,在植物对生物和非生物胁迫以及发育过程的反应中发挥着各种作用。本研究通过计算机克隆和反转录 PCR(RT-PCR),从受小麦条锈菌侵染的小麦叶片(品种 Suwon 11)中分离并鉴定了一个钙结合蛋白基因,命名为 TaCab1(小麦钙结合 EF 手蛋白 1)。TaCab1 没有内含子,预测编码一个 216 个氨基酸的蛋白质,其具有 N 端信号肽、跨膜结构域、EF 手模体和钙调素结构域。利用 TaCab1 与绿色荧光蛋白(GFP)基因的瞬时表达构建体的结果表明,TaCab1 编码一种跨膜蛋白。实时定量 PCR(qRT-PCR)分析表明,TaCab1 在叶片中的表达水平明显高于根和茎。尽管 TaCab1 在不亲和和亲和互作中的上调表达模式非常相似,但在亲和互作中其转录本的积累量明显高于不亲和互作。用植物激素[脱落酸、苄基腺嘌呤、乙烯、茉莉酸甲酯和水杨酸(SA)]和应激刺激[创伤、低温、聚乙二醇和高盐]处理后,TaCab1 的转录也被不同程度地上调。这些结果表明,TaCab1 通过 SA 信号通路参与植物-病原体识别、症状发育以及对生物和非生物胁迫的基础耐受性。

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