欢迎访问《中国农学通报》,

中国农学通报 ›› 2018, Vol. 34 ›› Issue (6): 137-142.doi: 10.11924/j.issn.1000-6850.casb17020079

所属专题: 生物技术 园艺

• 生物技术科学 • 上一篇    下一篇

利用分子标记鉴定白菜细胞质雄性不育类型的研究

兰梅,胡靖锋,刘家佳,张丽琴,杨红丽,徐学忠,姚文勇,和江明   

  1. 云南省农业科学院园艺作物研究所,云南省农业科学院园艺作物研究所,云南省农业科学院园艺作物研究所,云南省农业科学院园艺作物研究所,云南省农业科学院园艺作物研究所,云南省农业科学院园艺作物研究所,云南省大理州弥渡县农业技术推广中心,云南省农业科学院园艺作物研究所
  • 收稿日期:2017-02-21 修回日期:2018-01-30 接受日期:2017-04-19 出版日期:2018-02-26 发布日期:2018-02-26
  • 通讯作者: 和江明
  • 基金资助:
    国家大宗蔬菜产业技术体系“国家现代农业产业技术体系大宗蔬菜昆明综合试验站”(CARS-23-G37)。

Identifying Cytoplasmic Male Sterile Types of Chinese Cabbage (Brassica campestris L.) by Molecular Markers

  • Received:2017-02-21 Revised:2018-01-30 Accepted:2017-04-19 Online:2018-02-26 Published:2018-02-26

摘要: 通过对9份白菜细胞质雄性不育材料的不育胞质类型进行分子标记鉴定,以期为今后更好的利用分子标记来进行辅助育种奠定基础。通过CTAB法提取白菜的基因组DNA,根据GenBank中orf138基因保守序列设计2对特异引物,对9份白菜基因组DNA进行PCR扩增,将扩增出条带的PCR产物进行测序,在NCBI中进行同源性比对,最终确定其不育胞质类型。结果表明:2对特异引物PI/PII和PIII/PIV只在4份白菜雄性不育基因中均扩增出条带,在另外5份材料中未扩增出条带,与田间育性鉴定相符,获得4份白菜Ogura胞质雄性不育的分子标记;在GenBank中用BLAST进行同源性比对分析,发现L1-CI、L3-CI、L3-F1 3个不育材料的特异片段与已报道的青花菜Ogu CMS所具有的Ogu orf138基因(Genbank登录号:HQ149728)同源度高达100%,L1-F1的同源度达99%,出现了一个变异位点,另外5份材料的胞质不育类型有待进一步研究。通过该方法鉴定出9份材料中有4份材料是萝卜胞质雄性不育的材料,得到了4条分子标记,该结果可为白菜细胞质雄性不育的分子鉴定提供了新的标记,也为今后利用分子标记辅助育种奠定了基础。

关键词: 黄瓜疫霉菌, 黄瓜疫霉菌, 遗传多样性, 系统发育树, ITS, β-tubulin, 序列分析

Abstract: Different sterile cytoplasm types of nine cabbage cytoplasmic male sterile materials were identified by molecular marker in the study in order to better use molecular marker to conduct the assisted breeding in the future. Genomic DNA was isolated from Chinese cabbage by CTAB method. The design of two pairs of specific primers was performed on conserved flanking region of orf138 gene in the GenBank. PCR were performed with genomic DNA of nine Chinese cabbage materials. The bands were sequenced. The homologous comparison was conducted in NCBI, and finally, the type of sterile cytoplasm was determined. The results showed that the bands were amplified only in four Chinese cabbage male sterile materials with two pairs of specific primers PI/PII and PIII/PIV, while the other five materials did not obtain the relative bands. The result was consistent with the field sterility identification. And then four molecular markers of Chinese cabbage Ogura cytoplasmic male sterility (CMS) were obtained. After conducting a homologous comparative analysis with BLAST in GenBank, it was found that the homologous degree was 100% in specific segments of the three sterility materials (L1-CI, L3-CI and L3- F1) and Oguorf138 gene (GenBank accession No.: HQ149728) of the reported broccoli Ogu CMS. The homologous degree of L1-F1 was 99% with a variation point. The type of cytoplasmic male sterility of the other five materials needed further research. Four materials of the nine were identified as the radish cytoplasmic male sterility materials and four molecular markers were obtained.

中图分类号: