水稻OsMPK15的cDNA克隆和转录水平分析

doi:10.13560/j.cnki.biotech.bull.1985.2017-0978

摘要/Abstract

摘要： MAPK基因参与水稻的生物与非生物胁迫应答及生长发育过程，通过克隆水稻OsMPK15并初步研究其表达特性，为后续功能研究奠定基础。采用RT-PCR技术从日本晴水稻的根组织中克隆OsMPK15的cDNA编码区，并采用实时荧光定量PCR（qRT-PCR）技术分析OsMPK15在不同组织中以及不同非生物胁迫下的表达特性。结果表明，从日本晴水稻根组织中克隆的OsMPK15的cDNA编码区序列与生物信息学预测的结果一致，OsMPK15的第13-304位为丝氨酸/苏氨酸激酶结构域，属于水稻MAPK家族E组。qRT-PCR结果表明，该基因在水稻幼穗期各组织表达存在差异，主要在叶和叶鞘中表达。干旱和盐胁迫均可诱导OsMPK15在水稻幼苗根中表达量的上调，但表达模式具有不同的规律。同时，OsMPK15对3种胁迫相关激素ABA、JA和SA处理均产生应答，其中以ABA诱导OsMPK15表达上调幅度最大。

关键词: 水稻, OsMPK15, 转录水平表达特性, 非生物胁迫, 植物激素

Abstract: MAPK gene involves in the rice’s responses to biotic and abiotic stress as well as its development and growth. Cloning rice OsMPK15 and preliminarily studying its expression would provide foundation for further its functional studies. In this study，RT-PCR was used to clone its cDNA coding region of OsMPK15 from rice Nipponbare roots，and quantitative real time PCR（qRT-PCR）to analyze the expression characteristics in various tissues and under different abiotic treatments. The results showed that the cDNA coding sequence of OsMPK15 isolated from rice Nipponbare root was identical with the results of bioinformatics prediction. Sequence analysis revealed that a highly conserved serine/threonine kinase domain in MAPK protein was located at the position from 13th to 304th in OsMPK15 amino acids sequence，and it belonged to group E of MAPK family. qRT-PCR results demonstrated that OsMPK15 gene was widely expressed in various tissues，especially accumulated in leaves and leaf sheaths at young panicle stage. The expression of OsMPK15 gene in seedling roots was up-regulated by drought and salt stress with different expression pattern. Meanwhile，the treatment with stress-related hormones ABA，JA and SA resulted in the responses and the highest up-regulation by ABA.

Key words: rice, OsMPK15 gene, transcriptional expression analysis, abiotic stress, plant hormones

石佳, 杨丹丹 ,葛慧雯 ,杜京尧 ,梁卫红. 水稻OsMPK15的cDNA克隆和转录水平分析[J]. 生物技术通报, 2018, 34(6): 66-72.

SHI Jia YANG Dan-dan GE Hui-wen DU Jing-yao LIANG Wei-hong. cDNA Cloning and Transcriptional Expression Analysis of OsMPK15 in Rice[J]. Biotechnology Bulletin, 2018, 34(6): 66-72.

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