遗传 ›› 2009, Vol. 31 ›› Issue (5): 540-545.doi: 10.3724/SP.J.1005.2009.00540

• 研究报告 • 上一篇    下一篇

落叶松体细胞胚成熟阶段差异表达的基因及部分基因的表达谱分析

张蕾1, 2;齐力旺1;韩素英3   

  1. 1. 中国林业科学研究院林业研究所细胞生物学实验室, 北京 100091;
    2. 国家知识产权局专利审查协作中心, 北京100193;
    3. 中国林业科学研究院森林生态环境与保护研究所, 北京 100091
  • 收稿日期:2008-06-24 修回日期:2009-02-25 出版日期:2009-05-10 发布日期:2009-05-10
  • 通讯作者: 齐力旺

Differentially expressed genes during Larix somatic embryo maturation and the expression profile of partial genes

ZHANG Lei1, 2;QI Li-Wang1;HAN Su-Ying3   

  1. 1. Laboratory of Cell Biology, Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, China;
    2. Patent Examination Cooperation Center, State Intellectual Property Office of the People’s Republic of China, Beijing 100193, China;
    3. Institute of Forest Ecology, Environment and Protection, Chinese Academy of Forestry, Beijing 100091, China
  • Received:2008-06-24 Revised:2009-02-25 Online:2009-05-10 Published:2009-05-10
  • Contact: QI Li-Wang

摘要:
摘要: 为研究落叶松体细胞胚胎发生的分子机理, 文章以日本落叶松×华北落叶松杂种无性系胚性细胞系Y35体细胞胚成熟阶段培养物的cDNA为实验组, 继代培养阶段胚性愈伤组织的cDNA为对照组, 利用抑制性消减杂交技术(Suppression subtractive hybridization, SSH)构建了体细胞胚成熟阶段的差异表达基因文库。随机选取800个阳性克隆进行测序, 共获得468个UniGenes, 共将其分为19类, 功能分析结果表明: 这些UniGenes可能参与代谢、转录、信号转导、转运、细胞生长分裂、细胞结构、细胞命运、蛋白质合成与降解、防御等与个体发育密切相关的生物学过程。对部分ESTs的表达谱进行分析, 结果表明这些ESTs均在落叶松体细胞胚胎发生的不同阶段特异表达。

关键词: 差异表达基因, 抑制性消减杂交, 体细胞胚成熟, 落叶松

Abstract: Abstract: To study the molecular mechanism of Larix somatic embryogenesis, a differentially expressed cDNA library of Larix somatic embryo in the period of maturation was constructed using suppression subtractive hybridization (SSH). The cDNA from the cultures at the stage of somatic embryo maturation of embryogenic cell line Y35 of L. leptolepis ×L. princi-pis-rupprechtii was used as the tester and the cDNA from its subcultured callus was used as the driver. Eight hundreds ran-domly selected positive clones were sequenced, and 468 UniGenes were obtained finally. According to their function, these ESTs were classified into 19 categories and were involved in many biological process related to plant growth and develop-ment such as metabolism, transcription, signal transduction, transport facilitation, cell growth and division, cell structure, cell fate, protein synthesis or degradation, defense etc. Real-time PCR results of several ESTs showed that they were all differentially expressed at the different stages during cell line Y35 somatic embryo maturation.