东乡野生稻MYB转录因子LOC_Os01g62410-DX苗期低温诱导荧光定量表达分析及克隆  

Cloning and Quantitative Expression of MYB Transcription Factor LOC_Os01g62410-DX from Dongxiang Wild Rice at Seedling Stage under Low Temperature Stress

作　　者：於紫蕾 却志群[1] 沈春修[1] YU Zilei;QUE Zhiqun;SHEN Chunxiu(College of Life Sciences,Resources and Environment Sciences,Yichun University,Jiangxi Key Laboratory of Crop Growth and Development Regulation,Yichun 336000, China)

机构地区：[1]宜春学院生命科学与资源环境学院,江西省作物生长发育调控重点实验室,江西宜春336000

出　　处：《西北农业学报》2020年第2期207-215,共9页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：国家自然科学基金项目(31660379);江西省教育厅科学技术研究项目(GJJ180845);江西省作物生长发育调控重点实验室开放课题项目(KFJJ201804)。

摘　　要：为了阐明东乡野生稻中MYB转录因子基因LOC_Os01g62410-DX在低温胁迫条件下的功能作用机制,依据课题组前期的转录组分析数据,以东乡野生稻和籼稻品种“93-11”为材料,通过实时荧光定量PCR分析该基因位点在冷胁迫条件下的表达模式。并参考公共数据库中测序水稻品种日本晴对应位点的序列信息设计引物,利用RT-PCR技术,从东乡野生稻中扩增获得LOC_Os01g62410-DX基因的全长cDNA,构建该基因的过表达载体。序列分析结果表明,LOC_Os01g62410-DX基因的cDNA全长为1500 bp,共编码499个氨基酸,蛋白质分子质量为55.04 ku,理论等电点(pI)为7.62。在线分析软件预测LOC_Os01g62410-DX氨基酸序列N端包含2个高度保守结构域SANT,表明该基因属于R2R3类MYB转录因子。系统进化分析结果表明LOC_Os01g62410-DX与水稻Osmyb3和Osmyb4编码的蛋白位于邻近分支上,亲缘关系最近。进一步利用农杆菌介导转基因方法,获得转入LOC_Os01g62410-DX基因的37株阳性转基因植株,为深入研究东乡野生稻LOC_Os01g62410-DX基因在低温胁迫条件下的作用机制奠定材料基础。In order to explore the functional mechanism of MYB transcription factor LOC_Os01g62410-DX from Dongxiang wild rice under low temperature stress,the expression analysis of LOC_Os01g62410-DX was performed by means of fluorescence quantitative PCR technology based on previous transcription data.The full-length cDNA of LOC_Os01g62410-DX gene was amplified from Dongxiang wild rice using reverse transcription polymerase chain reaction(RT-PCR)with the primers designed on the basis of corresponding gene locus sequence information in the public database,and its over-expression vector was then constructed.Sequence analysis showed that the full-length cDNA of LOC_Os01g62410-DX was 1500 bp and encoded a protein about 55.04 KD,which was composed of 499 amino acids and corresponding theoretical isoelectric point(pI)was 7.62.Online analysis software prediction showed that the N-terminal amino acid sequence of LOC_Os01g62410-DX contained two highly conserved structure domain SANT.It was suggested that the gene was a member of the R2R3 MYB transcription factors family.Phylogenetic analysis results showed that the LOC_Os01g62410-DX had the closest relationship with Osmyb3 and Osmyb4 protein in rice.LOC_Os01g62410-DX was then transformed into the receptor rice variety TP309 via Agrobacterium mediated transformation,and 37 positive transgenic plants were obtained after hygromycin selection and GUS histochemical staining detection.This study laid the important material base for gene function research under low temperature stress.

关 键 词：东乡野生稻 冷胁迫 转录组 实时荧光定量 MYB转录因子 

分 类 号：S184[农业科学—农业基础科学] S330