甜瓜α-甘露糖苷酶亚家族基因的克隆及分析  

Cloning and Analysis of α-mannosidase Subfamily Genes in Melon(Cucumis melo L.)

作　　者：郝鑫[1] 田立莉 王琳博 哈斯阿古拉[1] 郝金凤[1] Hao Xin;Tian Lili;Wang Linbo;Hasi Agula;Hao Jinfeng(Inner Mongolia Key Laboratory of Herbage ＆ Endemic Crop Biotechnology,College of Life Sciences,Inner Mongolia University,Hohhot,010021)

机构地区：[1]内蒙古大学生命科学学院内蒙古自治区牧草与特色作物生物技术重点实验室

出　　处：《基因组学与应用生物学》2018年第9期3903-3908,共6页Genomics and Applied Biology

基　　金：国家自然科学基金项目(31101559);内蒙古大学校级创新训练项目(201514239)共同资助

摘　　要：在植物体中,α-甘露糖苷酶（α-man,α-mannosidase）是N-聚糖加工、修饰的关键酶,而N聚糖的加工对于植物生长发育是必不可少的,同时在果实成熟过程中起着重要作用。对甜瓜Ⅱ类α-甘露糖苷酶家族基因的结构特点和表达模式进行分析,为探讨α-甘露糖苷酶基因家族各成员在甜瓜不同组织器官发育中可能存在的作用奠定基础。本研究以甜瓜品种河套蜜瓜为材料,采用RT-PCR方法克隆了该家族的3个成员：Cm MAN1、Cm MAN2和Cm MAN3的全长c DNA,应用生物信息学方法对其相应蛋白质的理化性质、系统发育、保守基序进行了分析。分析研究表明,Cm MAN1、Cm MAN2和Cm MAN3的开放阅读框分别为3 063 bp、3 483 bp和3 024 bp,分别编码1 020、1 160和1 007个氨基酸,均为酸性蛋白质,且在不同物种间具有高度保守性。利用RT-q PCR方法检测了这些基因的表达模式,发现Cm MAN1在成熟甜瓜果实中表达量最高,Cm MAN2在叶、花和子房中表达量相对较高,Cm MAN3在子房中表达量较高,表明α-甘露糖苷酶各基因在甜瓜发育中可能具有不同的作用。In plants, α-mannosidase is a key enzyme in the processing and modification of of N-glycan, which is essential for plant growth and development, and plays an important role in fruit ripening. The analysis of structure and expression pattern of α-mannosidase subfamily genes could lay the foundation for the functional exploration of these genes in the growth and development of different tissues and organs in melon（Cucumis melo L. cv Hetao）.In this study, the full-length c DNAs of three genes in α-mannosidase subfamily, Cm MAN1, Cm MAN2, and Cm MAN3, were cloned by RT-PCR. The physicochemical properties, phylogeny, and conservative motif of the corresponding proteins were analyzed by bioinformatics. The results showed that the open reading frames of Cm MAN1, Cm MAN2, and Cm MAN3 were 3 063 bp, 3 483 bp, and 3 024 bp, and encoded 1 020, 1 160, and 1 007 amino acids, respectively. These proteins were all acidic proteins and showed highly conservatism among different species. The expression patterns of these genes were detected by RT-q PCR method. It was found that the expression of Cm MAN1 was the highest in mature melon fruit, that of Cm MAN2 was relatively high in leaves, flowers,and ovaries, and that of Cm MAN3 was relatively high in ovaries. These results indicated that α-mannosidase genes could play different roles in melon development.

关 键 词：甜瓜 α-甘露糖苷酶 序列分析 表达特性 

分 类 号：S652[农业科学—果树学]