小麦耐热相关转录因子基因TabZIP28的分离及功能分析

发布时间：2018-05-17 14:24

本文选题：小麦 + b　；参考：《农业生物技术学报》2016年02期

【摘要】：碱性亮氨酸拉链(basic leucine zipper,b ZIP)是植物中广泛存在的一类转录因子,参与多种胁迫响应与生长发育过程。本研究从小麦(Triticum aestivum)中克隆到一个热胁迫诱导的b ZIP家族转录因子基因Tab ZIP28(Gen Bank登录号:KT753298.1),ORF长度为1 713 bp,编码570个氨基酸。生物信息学分析结果表明,Tab ZIP28与拟南芥b ZIP家族转录因子中B亚组的3个基因Atb ZIP17、Atb ZIP28和Atb ZIP49归为一类。氨基酸序列比对结果表明,该蛋白具有b ZIP和跨膜结构域(transmembrane domain,TMD)两个保守结构域以及规范的位点1蛋白酶(site 1 protease,S1P)剪切位点。对该基因起始密码子ATG上游1 699bp的序列进行顺式作用元件分析,发现该基因的启动子区域包含众多激素和逆境胁迫响应元件。通过q RT-PCR对该基因在逆境胁迫下的表达模式进行分析,结果表明,Tab ZIP28在热胁迫处理1 h即上调表达且达到最大值;用20%PEG 6000模拟干旱环境处理小麦幼苗后,Tab ZIP28在处理6 h达到最大值,并在12 h时急剧下降;对5 mmol/L H2O2处理响应比较缓慢,在处理12 h才上调表达;该基因不受到二硫苏糖醇(dithiothreitol,DTT)处理的诱导表达。在拟南芥(Arabidopsis thaliana)中过量表达Tab ZIP28基因,转基因株系在高温胁迫后的成活率和种子发芽率较野生型明显提高,说明该基因可能对植物的耐热性有贡献,可以作为耐热性育种的候选基因。
[Abstract]:Alkaline leucine zipper basic leucine zipperb (ZIP) is a widespread transcription factor in plants, which is involved in many stress response and growth and development processes. In this study, a heat stress induced b ZIP family transcription factor gene, Tab ZIP28(Gen Bank accession number, was cloned from Triticum aestivum. The length of Tab ZIP28(Gen Bank was 1 713 BP, encoding 570 amino acids. Bioinformatics analysis showed that Tab ZIP28 and three genes of B subgroup Atb ZIP17Ab ZIP28 and Atb ZIP49 in Arabidopsis thaliana B ZIP family were classified into one group. The results of amino acid sequence alignment showed that the protein had two conserved domains, b ZIP and transmembrane domain (TMD), as well as the normal site 1 protease S1P. The sequence of 1 699bp upstream of the initiation codon of the gene was analyzed by cis-acting element analysis. It was found that the promoter region of the gene contained many hormones and stress response elements. Q RT-PCR was used to analyze the expression pattern of the gene under stress. The results showed that Tab ZIP28 was up-regulated at 1 h of heat stress and reached its maximum value. 20%PEG 6000 was used to simulate the drought environment of wheat seedlings, and Tab ZIP28 reached its maximum value at 6 h and decreased sharply at 12 h, and the response to 5 mmol/L H2O2 treatment was slow, and the expression of Tab ZIP28 was up-regulated at 12 h after treatment. The gene was not induced by dithiothreitol DTT treatment. The overexpression of Tab ZIP28 gene in Arabidopsis thaliana of Arabidopsis showed that the survival rate and seed germination rate of transgenic lines after high temperature stress were significantly higher than that of wild type, which suggested that the gene might contribute to the heat tolerance of plants. It can be used as a candidate gene for heat resistance breeding.
【作者单位】：中国农业大学农学院/植物遗传育种学系/农业生物技术国家重点实验室/北京市作物遗传改良重点实验室;
【基金】：国家自然科学基金(No.31171549) 转基因专项(No.2011ZX08002-002)
【分类号】：S512.1

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