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
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
复制标题
小麦钙结合 EF-hand 蛋白基因 TaCab1 的克隆和表征及其响应条锈菌 f 的表达。
DOI：
10.1007/s11033-010-0501-8
复制
发表时间：
2011-08-01
影响因子：
2.8
通讯作者：
Kang, Zhensheng
中科院分区：
文献类型：
--
作者：
Feng, Hao;Wang, Xiaomin;Kang, Zhensheng
钙离子是真核生物信号转导途径中普遍存在的重要第二信使。钙结合蛋白是植物体内钙代谢途径的一个组成部分，在植物对生物和非生物胁迫的应答以及发育过程中发挥着重要作用。在这项研究中，钙结合蛋白基因，命名为TaCab 1（小麦钙结合EF手蛋白1），从小麦叶片（cv.水原11）感染条锈菌（Puccinia striiformis f.）通过电子克隆和逆转录PCR（RT-PCR）技术对该菌株进行了初步鉴定。TaCab 1不含内含子，编码216个氨基酸的蛋白质，其N端含有信号肽、跨膜结构域、EF-手状结构域和caleosin结构域。利用带有绿色荧光蛋白（GFP）基因的TaCab 1构建体进行的瞬时检测的结果表明，TaCab 1编码跨膜蛋白。实时荧光定量PCR（qRT-PCR）分析表明，TaCab 1在叶片中的表达量高于根和茎。虽然TaCab 1的上调表达谱在不相容和相容相互作用中非常相似，但其在相容相互作用中的转录积累远高于不相容相互作用。TaCab 1基因在植物激素（脱落酸、苄基腺嘌呤、乙烯、茉莉酸甲酯和水杨酸）和胁迫（创伤、低温、聚乙二醇和高盐）处理后，其转录水平也有不同程度的上调。这些结果表明TaCab 1通过SA信号通路参与植物病原体识别、症状发展以及对生物和非生物胁迫的基本耐受性。
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.