四个苹果MdCBF基因克隆与表达分析  

Cloning and Expression Analysis of Four Apple Md CBF Genes

作　　者：靳芹 代培红 刘超 胡芹华 刘晓东 李月 JIN Qin;DAI Peihong;LIU Chao;HU Qinhua;LIU Xiaodong;LI Yue(College of Agronomy,Xinjiang Agricultural University,Key Lab of Agricultrual Biological Technology,Urumqi 830052,China)

机构地区：[1]新疆农业大学农学院,农业生物技术重点实验室,新疆乌鲁木齐830052

出　　处：《华北农学报》2017年第A01期17-22,共6页Acta Agriculturae Boreali-Sinica

基　　金：国家自然科学基金项目(31660433);新疆农业大学大学生创新项目

摘　　要：为了筛选出苹果中的抗冻负调控基因,培育抗冻苹果新品种奠定基础,通过生物信息学的方法从苹果中克隆了4个具有完整开放阅读框的CBF基因,命名为Md CBFL1~Md CBFL4。通过与其他植物CBF/DREB蛋白氨基酸序列的多重比对发现,这4个蛋白具有CBF蛋白典型的结构特点,即1个AP2结构域,2个特征序列,即核定位信号区PKRPAGRTKFRETRHP和DSAWR保守序列。从系统进化树中可以看出苹果Md CBFL1~Md CBFL4蛋白与At DREB1蛋白聚在A-1亚族,推测它与拟南芥A-1转录因子功能相似,Md CBFL1~Md CBFL4蛋白表达受干旱、盐碱、低温等逆境胁迫的诱导。采用半定量RT-PCR的方法检测分析苹果中Md CBFL1~Md CBFL4基因在高盐(200 mmol/L Na Cl)、干旱和低温处理下基因的表达情况,结果表明,Md CBFL1基因对低温、干旱、盐胁迫3种逆境胁迫均有一定程度的响应,Md CBFL2~Md CBFL4基因受低温与高盐的诱导。为进一步阐明苹果Md CBF基因在逆境应答中的功能、机制提供了有益线索。In order to select antifreeze negative gene in apple,lay a foundation for cultivating new varieties of antifreeze apples,4 open reading frame CBF genes were cloned from apple by the method of information biology,named as MdCBFL1-MdCBFL4.These genes(MdCBFL1-MdCBFL4)were shown to encode proteins that share a high similarity with plant cold stress-related CBF proteins,which contained a AP2 domain and two classic sequences that conserved PKRPAGRTKFRETRHP nuclear location signal area and DSAWR conservative sequence.Phylogenetic analysis revealed that the isolated apple genes could be classified into the Arabidopsis A-1 subgroup,it was speculated that it was similar to the function of Arabidopsis A-1 transcription factor,and the expression of MdCBFL1-MdCBFL4 proteins was induced by stress of drought,low temperature and salt stress.The expression of MdCBFL1-MdCBFL4 gene in apple was detected by semi-quantitative RT-PCR in high salt(200 mmol/L NaCl),drought and low temperature.The results showed that MdCBFL1 gene was response to the low temperature,drought and salt stress.MdCBFL2-MdCBFL4 gene was induced by low temperature and high salt.This provides useful clues to further elucidate the function and mechanism of apple MdCBF gene in stress response

关 键 词：苹果 CBF DREB转录因子 基因克隆 非生物胁迫 表达分析 

分 类 号：Q78[生物学—分子生物学] S661.03[农业科学—果树学]