陆地棉GhPYR1基因的克隆和功能分析

doi:10.13560/j.cnki.biotech.bull.1985.2016.02.012

摘要/Abstract

摘要： 植物激素脱落酸(Abscisic acid，ABA)在植物应对干旱、盐碱等逆境胁迫以及植物种子萌发、根伸长、芽休眠等阶段发挥重要作用。PYR/PYL/RCAR蛋白家族是ABA受体，与ABA结合后能够启动ABA信号传导通路，诱导ABA应答基因的表达。利用电子克隆和RT-PCR技术从陆地棉中克隆了GhPYR1基因，其编码的GhPYR1蛋白与拟南芥中AtPYR1蛋白相似度为73%。将GhPYR1蛋白序列与拟南芥14个PYR/PYL/RCAR家族成员蛋白序列进行比对并构建进化树，发现它与拟南芥PYR/PYL/RCAR蛋白亚家族III亲缘关系最近。过表达GhPYR1基因的T₃代拟南芥在外源ABA处理下，其种子萌发和初期根生长均滞后于野生型，表现出对ABA更加敏感；高盐和干旱胁迫对转基因种子的萌发抑制更强烈，但苗期胁迫处理下转基因拟南芥的长势却明显优于野生型；同时在外源ABA诱导条件下ABA应答基因RD29A、RAB18的表达量较野生型有明显提高。以上结果说明GhPYR1基因编码的蛋白是ABA的受体，过表达该基因能够提高植物对ABA的敏感性和增强应对逆境胁迫的能力。

关键词: 陆地棉, GhPYR1基因, 生物信息学分析, 生理功能分析

Abstract: The phytohormone abscisic acid(ABA)plays an important role in response to drought, salinity and other abiotic stress.It is also a key regulator involved in different plant developmental stages such as seed germination, root elongation, and bud dormancy.The PYR/PYL/RCAR protein family is the ABA receptor, which can initiate the ABA signaling pathway and induce the expression of ABA response genes.In this study, gene GhPYR1 was cloned from Gossypium hirsutum L.with electronic cloning and RT-PCR technology.GhPYR1 protein exhibited 73% sequence identity with Arabidopsis AtPYR1 protein.Protein sequence alignment among GhPYR1 and 14 family members of PYR/PYL/RCAR in Arabidopsis as well as the construction of phylogenetic tree revealed that gene GhPYR1 was most closely related to subfamily III of PYR/PYL/RCAR protein in Arabidopsis.T₃ generation Arabidopsis over-expressing gene GhPYR1were more sensitive to exogenous ABA, as its seed germination and early root growth lagged behind than wild type.The germination of transgenic seeds was more strongly inhibited by high salt and drought stress, however, the transgenic lines were significantly better than the wild ones in the seeding stage.Moreover, the ABA-responsive gene RD29A and RAB18 in transgenic Arabidopsis were significantly induced by ABA compared to the wild type.The above results show that the protein encoded by gene GhPYR1 is the receptor of ABA and over-expression of GhPYR1 can improve the sensitivity of plant to ABA and enhance the ability to respond to stress in plants.

Key words: Gossypium hirsutum L. , GhPYR1 gene, bioinformatics analysis, physiological function analysis

刘妍, 孟志刚, 孙国清, 王远, 周焘, 郭三堆, 张锐. 陆地棉GhPYR1基因的克隆和功能分析[J]. 生物技术通报, 2016, 32(2): 90-99.

LIU Yan, MENG Zhi-gang, SUN Guo-qing, WANG Yuan, ZHOU Tao, GUO San-dui, ZHANG Rui. Cloning and Function Analysis of Gene GhPYR1 in Gossypium hirsutum L.[J]. Biotechnology Bulletin, 2016, 32(2): 90-99.

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