草业学报2017，Vol.26Issue(12)：108-116,9.DOI:10.11686/cyxb2017202
紫花苜蓿MsLEA4-4基因的克隆、表达分析及遗传转化
Cloning and expression analysis of MsLEA4-4 from Medicago sativa
摘要
Abstract
Late embryogenesis abundant (LEA) proteins are widely involved in many adverse situations for plants.In this study,we isolated a LEA4 gene from alfalfa by homology cloning strategy,termed MsLEA4-4.The MsLEA4-4 encoding revealed 512 amino acids and structure analysis showed that the gene contains eleven repeating TAQAAKEKTQQ amino acids sequences.Quantitative real-time PCR was used to detect MsLEA4-4 expression quantities in adverse situations.Results revealed that the expression of MsLEA4-4 was up-regulated by drought (PEG),salinity (NaCl),Cu2+,Zn2+ and ABA stress.Gene expression was highest at 2 h and 8 h under the stress of NaCl,Cu2+ and Zn2+.Under chilling and drought stress,the gene showed a rising trend.These results indicate that the MsLEA4-4 protein may be involved in the regulation of environment stress responses in alfalfa.The plant expression vector pCAMBIA3301-MsLEA4-4 was constructed and transgenic plants were obtained through infecting inflorescences of Arabidopsis by Agrobacterium tumefaciens-mediated transformation.The vector sequence was tested by antibiotic (PPT) and PCR in the genome of transgenic plants and indicated that the target gene had been transferred.This study provides basic information for further study of the MsLEA4-4 function in stress-tolerance regulation in alfalfa.关键词
紫花苜蓿/胚胎晚期富集蛋白(LEA)/逆境胁迫/遗传转化Key words
Medicago sativa/late embryogenesis abundant proteins (LEA)/adversity stress/genetic transformation引用本文复制引用
贾会丽,王学敏,郭继承,高洪文,吴欣明,刘建宁,石永红,董宽虎,王运琦..紫花苜蓿MsLEA4-4基因的克隆、表达分析及遗传转化[J].草业学报,2017,26(12):108-116,9.基金项目
现代农业产业技术体系专项资金(CARS-35-25),物种资源保护费(2130135)和山西省农业科学院畜牧兽医研究所项目(xms201506)资助. （CARS-35-25）
