小麦TaZTP29基因的克隆及耐盐性鉴定

发布时间：2018-02-03 09:02

  本文关键词： 小麦 TaZTP基因 耐盐 基因表达 亚细胞定位　出处：《农业生物技术学报》2017年10期 　论文类型：期刊论文

【摘要】：土壤盐碱化是小麦(Triticum aestivum)生产的主要限制因子之一。锌转运体(zinc transporter,ZTP)属于锌铁调控蛋白(ZRT,IRT-like protein,ZIP)蛋白家族,并参与调控非折叠蛋白应激响应,是一个优良的抗盐基因。本研究采用同源克隆的方法获得了ZTP29在小麦中的直系同源基因TaZTP29,利用生物信息学、qRT-PCR、绿色荧光蛋白示踪技术和转基因拟南芥(Arabidopsis thaliana)过表达的方法明确了TaZTP29的保守域信息、表达模式、亚细胞定位信息和功能。序列分析结果表明,TaZTP29基因(GenBank登录号:KY610283)含有81 bp的5'UTR、834 bp基因编码区及117 bp的3'UTR,编码277个氨基酸。TaZTP29蛋白具有典型的ZIP保守域,具有8个跨膜域,且在第V跨膜域具有完全保守的组氨酸蛋白残基,定位于质膜上,属于ZIP蛋白家族。序列同源性和进化分析表明,TaZTP29与节节麦(Aegilops tauschii)ZIP29蛋白同源性最高;与拟南芥(Arabidopsis thaliana)ZTP29蛋白同源性最远。TaZTP29强烈响应锌和盐胁迫,上调表达,在高温与干旱胁迫下,下调表达;且TaZTP29主要在地下部分的根中表达,在地上部分的组织中表达量较低。抗盐性鉴定表明,在不加NaCl的条件下,野生型与过表达株系总根长基本一致,在100和150 mmol/L NaCl处理的培养基中,3个转基因株系的总跟长显著长于野生型拟南芥,表明过表达株系对NaCl胁迫具有较好的耐性。转入TaZTP29后,在拟南芥中过表达能够提高拟南芥对盐胁迫的抗性,TaZTP29具有抗盐功能,为进一步改良小麦抗逆性等分子机理研究提供了新的依据。
[Abstract]:Soil salinization is one of the main limiting factors for Triticum aestivum production. ZTP belongs to the ZIP family of zinc-ferritin regulatory protein ZRTT-like protein and participates in the regulation of non-folding protein stress response. TaZTP29, a homologous gene of ZTP29 in wheat, was obtained by homologous cloning and bioinformatics qRT-PCR. The green fluorescent protein tracing technique and the method of overexpression of Arabidopsis thaliana in transgenic Arabidopsis confirmed the conserved domain information and expression pattern of TaZTP29. Subcellular location information and function. Sequence analysis showed that TaZTP29 gene (GenBank accession number: KY610283) contained 81bp 5UUTR. The coding region of 834bp gene and the 117bp 3G UTRs encode 277 amino acids. TaZTP29 protein has typical conserved ZIP domain and 8 transmembrane domains. And in the V transmembrane domain there is a completely conserved histidine protein residues located on the plasma membrane belonging to the ZIP protein family. Sequence homology and evolution analysis showed. TaZTP29 had the highest homology with Aegilops tauschii)ZIP29. TaZTP29, the farthest homology of Arabidopsis thaliana)ZTP29 protein from Arabidopsis, strongly responded to zinc and salt stress and upregulated the expression. Under high temperature and drought stress, the expression was down-regulated. The expression of TaZTP29 was mainly in the roots of the underground part, but low in the tissue of the aboveground part. The salt tolerance test showed that the expression of TaZTP29 was not under the condition of NaCl. The total root length of wild type and over-expressed strain was basically the same. In the medium treated with 100 and 150 mmol/L NaCl, the total heel length of three transgenic lines was significantly longer than that of wild type Arabidopsis thaliana. The results showed that the overexpression lines had better tolerance to NaCl stress, and overexpression in Arabidopsis thaliana could enhance the resistance of Arabidopsis thaliana to salt stress, TaZTP29 had the function of salt resistance. It provides a new basis for the further improvement of wheat stress resistance and other molecular mechanisms.
【作者单位】： 西北农林科技大学农学院;
【基金】：国家科技支撑计划项目(No.2013BAD04B02) 陕西省科技统筹创新项目(No.2014KZTB02-01-01) 中央高校基本科研业务费(No.Z109021423)
【分类号】：Q943.2;S512.1
【正文快照】： 小麦(Triticum aestivum)是我国第二大粮食作物,小麦的安全生产对保障我国的粮食安全具有重要意义(Zhang et al.,2016)。高温、干旱,特别是盐渍等非生物逆境已成为小麦产量和品质提高的主要限制因素。阐明植物高盐胁迫应答信号传导通路、克隆信号通路中的关键基因是利用生物技，

本文编号：1486965