柽柳 GF14基因的克隆与表达分析

生物体中普遍存在的14-3-3蛋白(也称GF14蛋白)能够参与植物的一系列应激响应过程。笔者以柽柳(Tamarix hispida)为材料,从6个柽柳cDNA文库中分离出5条GF14 基因,依次命名为ThGF14a—ThGF14e。对5条GF14 基因进行生物信息学研究,并利用qRT-PCR技术对胁迫处理后的基因表达模式进行分析。结果显示:ThGF14 蛋白除了N和C末端的氨基酸外均含有一个高度保守的14-3-3 结构域,这些ThGF14蛋白分属于ε-like 和 non-ε 两种类型; 大多数 ThGF14基因在NaCl、PEG、4 ℃、CdCl₂处理不同时间(6,24,48 和 72 h)的叶和根中能够被诱导表达(表达量>2倍),且不同基因间对非生物胁迫应答表现出差异,其中ThGF14c 基因在叶和根中不同胁迫条件下的表达水平均最高。研究表明,柽柳 ThGF14 基因能够参与植物的非生物胁迫应答响应过程。

Cloning and expression analysis of ThGF14 gene in Tamarix hispida

The ubiquitous family of 14-3-3(also known as GF14)proteins function as regulators in a variety of stress response processes in plants. Five GF14 genes from Tamarix hispida, designated ThGF14a to ThGF14 e, were identified from an exhaustive search of six cDNA library databases. The basic bioinformatics analysis of five GF14 genes were implemented, and the expression patterns of GF14 genes were analyzed by qRT-PCR under the treatment of stresses. The results were as follows: Bioinformatics analysis showed that ThGF14 proteins contained a highly conserved 14-3-3 domain, and the amino acid sequences are highly conserved except at the N- and C-terminal regions. A phylogenetic analysis indicated that ThGF14 proteins have both ε-like and non-ε forms. The expression profiles of the five ThGF14 genes in response to NaCl, polyethylene glycol(PEG), low temperature, and CdCl₂ for various treatment periods(6, 24, 48, and 72 h)in leaf and root tissues were investigated using qRT-PCR. The results showed that most of the five ThGF14 genes were induced(>2-fold)in leaf and root tissues under abiotic stress, and there were significant differences in response to stress between the various genes. Interestingly, the ThGF14c gene, which generated analogous results under stress in both tissues, was the most highly induced of expression level. Taken together, these results indicate that ThGF14 genes are integral to the abiotic stress response.