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林业科学 ›› 2013, Vol. 49 ›› Issue (9): 46-51.

• 论文与研究报告 • 上一篇    下一篇

油松雌雄球花高通量基因表达谱芯片分析

钮世辉1, 袁虎威1, 陈晓阳1,2, 李伟1   

  1. 1. 北京林业大学生物科学与技术学院 林木育种国家工程实验室 北京 100083;
    2. 华南农业大学林学院 广州 510642
  • 收稿日期:2013-01-14 修回日期:2013-03-07 出版日期:2013-09-25 发布日期:2013-09-19
  • 通讯作者: 李伟
  • 基金资助:

    国家自然科学基金项目(31370657)。

Microarray Analysis of Large Scale Gene Expression Profiles between Male and Female Cones of Pinus tabulaeformis

Niu Shihui1, Yuan Huwei1, Chen Xiaoyang1,2, Li Wei1   

  1. 1. National Engineering Laboratory for Tree Breeding College of Biological Sciences and Biotechnology, Beijing Forestry University Beijing 100083;
    2. College of Forestry, South China Agricultural University Guangzhou 510642
  • Received:2013-01-14 Revised:2013-03-07 Online:2013-09-25 Published:2013-09-19

摘要:

生殖调控是针叶树种遗传改良与良种生产过程中的重要环节,已经有许多实践经验积累,然而对其中雌雄球花发育及调控的分子生物学基础研究仍相当匮乏。以油松多个体多组织样本均一化cDNA文库为材料,通过转录组测序,获得46584个油松单基因簇;基于这些基因信息,设计并定制油松表达谱芯片;通过对雌雄球花基因表达芯片分析,筛选到2460个在球花发育后期显著差异表达的基因,其中1309个在雄球花优势表达,1151个在雌球花优势表达。分别从雄球花与雌球花优势表达的基因中各挑选1个表达水平差异超过800倍的基因,由于其功能未知,分别命名为MCE1(MaleConeExpressed1)与FCE1(FemaleConeExpressed1),并进一步分析它们在雌雄球花不同发育阶段的表达规律。结果表明:MCE1只在雄球花各发育阶段高水平表达,而在雌球花各发育阶段表达水平极低,推测其参与雄球花的整个发育过程;FCE1在早期雄球花中大量表达,但随着雄球花的发育其表达量会急剧降低,而在雌球花中却始终稳定地高水平表达,推测其在雌雄球花不同发育阶段发挥不同的作用。研究结果可为油松及近缘树种雌雄球花发育的分子生物学研究提供基础和新的思路。

关键词: 油松, 转录组测序, 基因芯片, 雌球花, 雄球花

Abstract:

Reproductive control is an important part of the conifer genetic improvement and seed production process, on which there are a lot of practical experience accumulation. However, the relevant study in molecular biology on the development and regulation of conifers male and female cones is still very scarce. The Pinus tabulaeformis transcriptome of a normalized cDNA library from multiple tissues and individuals was sequenced, and a total of 46 584 unigenes were found. Based on the information of these genes, microarrays of P. tabulaeformis were designed. Through the microarray analysis of large scale gene expression profiles between male and female cones at the later stage of development, 2 460 differentially expressed genes were obtained, of which 1 309 are predominantly expressed in male cones while 1 151 are predominantly expressed in female cones. Two unknown function genes with over 800 fold changes in their expression were chosen from the male and female and named as MCE 1 (Male Cone Expressed 1)and FCE 1 (Female Cone Expressed 1), respectively. Their expression patterns were analyzed in male and female cones at the different developmental stages. The results showed that the MCE 1 was specifically highly expressed in male cones, however almost not expressed in female cones at different developmental stages, indicating it could be involved in the development of male cones. The FCE 1 was highly expressed in male cones at the early stage, and the expression was drastically reduced with the development. However, the expression of the FCE 1 was always highly expressed in female cones at different developmental stages. Therefore, the FCE 1 may play a different role in the development of male and female cones. This study would provide a new method and facilitate future molecular biology studies on the development of male and female cones of P. tabulaeformis and other related species.

Key words: Pinus tabulaeformis, transcriptome sequencing, microarray, female cones, male cones

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