甜瓜乙烯受体基因Cm-ETR1 cDNA的克隆及表达特性分析  

Cloning and Expression Characterization of Ethylene Receptor Gene Cm-ETR1 from Melon (Cucumis melo L.)

作　　者：陈宇杰[1,2] 陈明[1] 乌兰巴特尔 郝金凤[1] 高峰[1] 哈斯阿古拉[1] 

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,呼和浩特010021 [2]内蒙古民族大学生命科学学院,通辽028000

出　　处：《生物技术通报》2013年第7期54-59,共6页Biotechnology Bulletin

基　　金：高等学校博士学科点专项科研基金资助课题(200801260002)

摘　　要：乙烯感知和信号转导的初始成分是乙烯受体,为探明甜瓜乙烯受体基因Cm-ETR1在甜瓜果实成熟过程中的作用,以甜瓜品种河套蜜瓜为材料,根据GenBank中登录的甜瓜乙烯受体基因Cm-ETR1的cDNA序列(登录号为AF054806),设计合成特异性引物,采用RT-PCR技术克隆得到Cm-ETR1基因全长cDNA序列,提交到GenBank中(登录号为EF495185)。序列分析表明,序列长度为2 256 bp,编码区为2 223 bp,编码740个氨基酸,与已报道的cantalupenis甜瓜ETR1基因的cDNA序列完全一致。Cm-ETR1蛋白的系统进化树分析结果表明,该乙烯受体蛋白在各物种间高度保守,与黄瓜乙烯受体蛋白相似性最高,一致性为99%,与龙眼乙烯受体蛋白相似性最低,一致性为86%。定量PCR分析结果显示,随着甜瓜果实内源乙烯合成量和成熟程度的增加,Cm-ETR1基因的表达量同步增加,在果实乙烯跃变期,Cm-ETR1的表达量也达到最高值,内源乙烯合成量与Cm-ETR1基因表达量间呈显著正相关,表明Cm-ETR1基因在甜瓜果实成熟过程中可能具有重要的作用。Ethylene receptor is the initial composition of ethylene perception and signal transduction. In order to investigate the function of melon ethylene receptor gene Cm-ETR1 during fruit ripening in melon, a pair of gene specific primers was designed based on the cDNA nueleotide sequence of Cm-ETR1 gene from melon （ accession number ： AF054806 ）,in GenBank. The full-length cDNA of Cm-ETR1 was cloned by RT-PCR from ripening fruit of melon （ Cucumis melo L. ev. Hetao ） , and was submitted to GenBank （ accession number ： EF495185 ） . Sequence analysis showed that the cloned cDNA was 2 256 bp long, contained an 2 223 bp ORF that encoded a polypeptide of 740 amino acids, and the cDNA sequence was consistent with that of reported melon variety cantalupenis ETR1 cDNA. Cm-ETRl protein phylogenetic tree analysis show that the ethylene receptor protein is highly conserved in all species, and Cm-ETR1 is highest similarity with cucumber ethylene receptor protein by 99% consistency, and lowest similarity of longan ethylene receptor protein by 86% consistency. Quantitative PCR analysis indicated that the expression level of Cm-ETR1 gene was increased, which coincided with the raise of the melon fruit ethylene production and fruit ripening process, and peaked at ethylene climacteric stage. The amount of endogenous ethylene had a positive correlation with the expression of Cm-ETRI gene. It reveals that Cm-ETR1 gene may play an important role in fruit ripening.

关 键 词：甜瓜 乙烯受体 Cm-ETR1 果实成熟 基因表达 

分 类 号：S652[农业科学—果树学]