互花米草BADH基因的克隆与定量表达分析  

Cloning and Quantitative Expression Analysis of BADH Gene from Spartina alterniflora

作　　者：刘振 张侠[1] 尹海波[1] 陈世华[1] 郭善利[1] LIU Zhen;ZHANG Xia;YIN Haibo;CHEN Shihua;GUO Shanli(College of Life sciences,Yantai University,Yantai 264005,China)

机构地区：[1]烟台大学生命科学学院,山东烟台264005

出　　处：《河南农业科学》2018年第11期45-49,共5页Journal of Henan Agricultural Sciences

基　　金：国家自然科学基金面上项目(31370296);烟台大学博士启动基金项目(TM09B25;HX10B23)

摘　　要：为研究甜菜碱醛脱氢酶(Bataine aldehyde dehydrogenase,BADH)基因的耐逆功能及分子机制,以盐生植物互花米草(Spartina alterniflora)为试验材料,利用RACE技术克隆了cDNA全长序列。生物信息学分析发现,该核苷酸序列全长1 983 bp,开放阅读框1 515 bp,编码504个氨基酸残基。Blast比对发现,该氨基酸与细叶结缕草、二穗短柄草、高粱氨基酸的相似性分别达到96%、87%、86%,且氨基酸数量基本相同。将该基因命名为Sa BADH。以tub为内参,对SaBADH基因在600 mmol/L的NaCl、20%的PEG处理下表达的定量分析表明,SaBADH受NaCl、PEG诱导表达,随着处理时间的延长,SaBADH基因的表达呈现先上升后下降的趋势,分别在处理24、12 h时达到最大值,分别约为对照的7、4倍,差异达到极显著水平,表明SaBADH基因可能在抵御盐和干旱对互花米草的胁迫中发挥重要作用。In order to study the inverse resistance function and molecular mechanism of BADH gene,the full-length cDNA of BADH gene was cloned from Spartina alterniflora by RACE strategy.Bioinformatic analysis showed that the full-length cDNA of BADH gene was 1 983 bp and the open reading frame was 1 515 bp which encoding 504 amino acids.Blast alignment found that amino acid sequence from Spartina alterniflora was similar to that of Zoysia tenuifolia,Brachypodium distachyon and Sorghum bicolor and shared 96%,87%and 86%identity,respectively.The number of amino acids were basically the same.So the gene was named SaBADH.Using tub as internal reference,the expression pattern of Spartina alterniflora under 600 mmol/L NaCl and 20%PEG treatments was studied.The results showed that SaBADH gene was induced by NaCl and PEG,suggesting that it may play an important role in salt and drought tolerance of Spartina alterniflora.With the prolongation of treatment time,the expression of SaBADH gene showed a trend of increasing first and then decreasing,reaching the maximum at 24 h,12 h,respectively,about 7 times and 4 times of the control,respectively,and the difference reached a very significant level.

关 键 词：互花米草 甜菜碱 BADH基因 基因克隆 抗旱耐盐性 

分 类 号：S45[农业科学—植物保护] S153