叶用芥菜细胞质雄性不育相关基因orf220的分子特性  

Molecular Identification of Cytoplasmic Male Sterility Associated Gene orf 220 in Leaf Mustard (Brassica juncea var. multiceps Tsen et Lee)

作　　者：杨景华[1] 张明方[1] 喻景权[1] 王涛[1] 陈竹君[1] 

机构地区：[1]浙江大学农业与生物技术学院园艺系

出　　处：《Acta Genetica Sinica》2005年第6期594-599,共6页

基　　金：国家自然科学基金(编号:30270907)资助~~

摘　　要：采用榨菜细胞质雄性系为不育源,通过变种间杂交和回交的方法,转育叶用芥菜细胞质雄性不育种质。根据Brassicanap型细胞质雄性不育相关基因orf222设计兼并引物,利用特异引物PCR方法,分别在不育源、F1、BC2以及BC3世代中扩增出一条600～700bp大小的特异条带,而在叶用芥菜保持系中无此条带,具细胞质基因遗传特性。进一步序列分析表明,此特异条带大小为663bp,具有起始密码子和终止密码子,编码220个氨基酸,定名为orf220。同时,orf220推导的氨基酸序列存在两个跨膜区,氨基端与Oenotheraberteriana中的COXⅢ(线粒体细胞色素氧化酶复合体亚基)、萝卜中的ATP8(线粒体ATP酶复合体亚基)以及向日葵中的ORFB(ATP酶复合体α亚基相关)蛋白氨基端高度同源。RT PCR方法分析表明,orf220基因的表达为组成性表达,无器官特异性。The cytoplasmic male sterility (CMS) was transferred into leaf mustard by inter varietal hybridization and subsequent backcrosses cms donor as using tuber mustard.Using specially designed primers,one specific band at 600~700 bp was distinctively amplified by PCR from cms plants of different backcrossed generations.It was genetically stable and cytoplasmically inheritable in the various backcrossed progenies.The sequencing result shows it is a 663 bp fragment with its own initiation and termination codons,presumably encoding for 220 amino acids (named orf 220).This deduced polypeptide had two trans membrane hydrophilic domains and its N terminus shared high similarity in amino acids constituents with COXⅢ protein from Oenothera berteriana ,ATP8 protein from radish and ORFC protein from sunflower.In addition,RT PCR analysis showed, orf 220 gene was constitutively expressed in leaves,roots and floral buds.However,the direct evidence of its involvement in expression of cms trait needs further confirmation.

关 键 词：叶用芥菜 细胞质雄性不育 orf220 

分 类 号：S637.2[农业科学—蔬菜学]