[Molecular identification of cytoplasmic male sterility associated gene orf 220 in leaf mustard (Brassica juncea var. multiceps Tsen et Lee)].

Jing-Hua Yang, Ming-Fang Zhang, Jing-Quan Yu, Tao Wang, Zhu-Jun Chen
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Abstract

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 COX III protein from Oenothera berteriana, ATP8 protein from radish and ORFB 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.

[芥菜(Brassica juncea var. multiceps Tsen et Lee)细胞质雄性不育相关基因orf220的分子鉴定]。
以块茎芥为供体,通过品种间杂交和回交将细胞质雄性不育转移到叶芥中。利用专门设计的引物,从不同回交世代的cms植株中扩增出600 ~ 700 bp的特异条带。它在各种回交后代中具有遗传稳定性和细胞质遗传性。测序结果显示,该片段长度为663 bp,有自己的起始和终止密码子,可能编码220个氨基酸(命名为orf220)。该多肽具有2个跨膜亲水结构域,其n端氨基酸组成与野花菜(Oenothera berteriana)的COX III蛋白、萝卜的ATP8蛋白和向日葵的ORFB蛋白高度相似。此外,RT-PCR分析显示,orf220基因在叶片、根和花蕾中均有组成性表达。但其参与cms性状表达的直接证据还有待进一步证实。
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