云南逸散紫花苜蓿MsVDE基因的克隆与表达分析  

Cloning and expression analysis of MsVDE in escaped alfalfa in Yunnan,China

作　　者：许超 何承刚[1] 姜华[1] XU Chao;HE Chenggang;JIANG Hua(College of Animal Science and Technology,Yunnan Agricuhural University,Kunming 650201,Yunnan,China;Yunnan Academy of Forestry and Grassland Sciences,Kunming 650201,Yunnan,China)

机构地区：[1]云南农业大学动物科学技术学院,云南昆明650201 [2]云南省林业和草原科学院,云南昆明650201

出　　处：《草业科学》2022年第6期1140-1150,共11页Pratacultural Science

基　　金：国家自然科学基金(31660682);云南省农业联合基金重点项目(202101BD070001-026);云南省技术创新人才(2018HB075)。

摘　　要：叶黄素循环是植物应对环境胁迫进行热耗散的光保护途径之一,紫黄质脱环氧化酶基因(VDE)是叶黄素循环脱环氧化过程的关键基因。本研究采用RACE技术首次克隆了云南逸散紫花苜蓿(Medicago sativa)的MsVDE基因,同时采用实时荧光定量分析干热胁迫下的MsVDE基因表达特征。结果表明,MsVDE全长1678 bp,CDS编码区1608 bp,编码535个氨基酸。氨基酸序列保守区域具有3个VDE基因的特征区,即N-端半胱氨酸富集区、脂质运载蛋白特征区和C-端谷氨酸富集区。进化树分析表明该基因与蒺藜苜蓿(Medicago truncatula)VDE基因的氨基酸序列亲缘关系最近。干热胁迫第4天和第8天时的MsVDE基因表达量分别比正常条件上调了2.13倍和1.97倍;喷洒二硫苏糖醇(1,4-Dithiothreitol,DTT)溶液后MsVDE基因表达量平均下调了55%。本研究初步得出,云南逸散紫花苜蓿MsVDE基因在调控叶黄素循环应对干热胁迫中起作用。The xanthophyll cycle of thermal dissipation is one of the photoprotective pathways for plants in response to environmental stresses.The VDE gene is crucial in the cyclic de-epoxidation of xanthophyll.In this study,the VDE gene of alfalfa(Medicago sativa;MsVDE)that had escaped in the Yunnan area of China was cloned for the first time using the RACE technique.Real-time fluorescence quantitative technology was also used to analyze characteristics of MsVDE expression under drought and heat stresses.Full-length MsVDE cDNA was successfully cloned from escaped alfalfa.The 1608 bp coding sequence specified 535 amino acids.Homologous sequence alignment showed that the MsVDE sequence of escaped alfalfa encoded three characteristic VDE gene family domains:The N-terminal cysteine-rich region,lipid carrier protein region,and C-terminal glutamate-rich region.Sequence alignment and phylogenetic analysis showed that MsVDE had the closest relationship with thistle alfalfa(M.truncatula).MsVDE expression was up-regulated 2.13 times and 1.97 times under drought and stresses compared with normal conditions on day 4 and day 8,respectively.MsVDE expression in leaves sprayed with dithiothreitol solution was down-regulated by 55%on average.The results show that the MsVDE gene of alfalfa that has escaped in the Yunnan area could play an important role in photoprotection and regulation of the xanthophyll cycle under drought and heat stresses.

关 键 词：干热胁迫 叶黄素循环 生物信息学 紫黄质脱环氧化基因 基因表达 RACE qPCR 

分 类 号：S541.9[农业科学—作物学]