中国农业科学 ›› 2009, Vol. 42 ›› Issue (10): 3706-3712 .doi: 10.3864/j.issn.0578-1752.2009.10.0041

• 研究简报 • 上一篇    下一篇

哈克尼西棉细胞质雄性不育系和保持系差异表达基因分析

马勇,吴建勇,邢朝柱,郭立平,巩养仓,崔明晖,王海林

  

  1. (中国农业科学院棉花研究所/农业部棉花遗传改良重点实验室)
  • 收稿日期:2009-01-10 修回日期:2009-03-06 出版日期:2009-10-10 发布日期:2009-10-10
  • 通讯作者: 邢朝柱

Differentially Expressed Genes Between Cytoplasmic Male Sterility Lines and Maintainer Lines of Gossypium harknessii

MA Yong, WU Jian-yong, XING Chao-zhu, GUO Li-ping, GONG Yang-cang, CUI Ming-hui, Wang Hai-lin
  

  1. (中国农业科学院棉花研究所/农业部棉花遗传改良重点实验室)
  • Received:2009-01-10 Revised:2009-03-06 Online:2009-10-10 Published:2009-10-10
  • Contact: XING Chao-zhu

摘要:

【目的】通过筛选哈克尼西棉细胞质雄性不育系和保持系间败育关键时期花药中差异表达的基因,为进一步揭示棉花雄性不育机理,更好地利用棉花细胞质雄性不育材料奠定基础。【方法】以哈克尼西棉细胞质雄性不育系和保持系间败育关键时期,即花粉母细胞时期之前的花药为研究材料,应用cDNA-AFLP技术分离和鉴定两材料间差异表达的片段,并对差异片段进行验证、克隆、测序和生物信息学分析。【结果】共得到108个差异表达片段,其中67个被成功克隆,通过BLAST分析发现,29个片段可以在NCBI数据库中找到同源序列,对这些序列进行Gene Ontology分析后得知这些片段主要参与信号转导、转录、能量代谢、细胞壁发育等相关过程。其中,在两材料间还分离到一些与RNA编辑和雄配子发育相关的基因存在差异表达,而这些可能与棉花细胞质雄性不育相关。【结论】本研究通过cDNA-AFLP的分析,在棉花细胞质雄性不育系和保持系间分离到了108个差异表达的片段,通过对这些片段的分析,对两材料间差异表达的基因信息有了一个基本了解,这些结果为更好地了解棉花细胞质雄性不育发生过程相关基因的调控网络,揭示棉花雄性不育机理打下基础。

关键词: 哈克尼西棉, 细胞质雄性不育, cDNA-AFLP, 基因表达, 不育机理

Abstract:

【Objective】 By identifying the differentially expressed transcripts of anthers at the critical period of abortion between cytoplasmic male sterility and maintainer lines of Gossypium harknessii, this study is to explore the possible molecular mechanism of the cytoplasmic male sterility, and lay a basis on utilizing cytoplasmic male sterility lines. 【Method】 cDNA amplified fragment length polymorphism analysis was used to isolate the differentially expressed fragments, these fragments were verified by reverse Northern blot, and then were cloned, sequenced and analyzed by bioinformatic tools. 【Result】 Totally, 108 differentially expressed fragments were identified, of which, 67 were succeeded in cloning. Homology searches performed by BLAST at web servers of NCBI showed that there were 29 fragments could be found homology sequences in GenBank database. By analyzing the Gene Ontology of these 29 sequences, it could be concluded that these sequences mainly participated in signal transducer, transcription, energy metabolism, cell wall development. Among them, some differentially expressed genes related to RNA edit and male gametogenesis were identified between two materials, and these maybe the mainl factors of male sterility. 【Conclusion】 The results may help to understand the relevant gene regulatory networks of cytoplasmic male sterility of cotton, and provide a pilot platform for further investigating the mechanism of CMS.

Key words: Gossypium harknessii, CMS, cDNA-AFLP, gene expression, molecular mechanism