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植物研究 ›› 2008, Vol. 28 ›› Issue (3): 293-298.doi: 10.7525/j.issn.1673-5102.2008.03.011

• 论文 • 上一篇    下一篇

白桦雌花序抑制性消减文库构建及EST分析

王 超1;杨传平1*;魏继承2;姜 静1   

  1. (1.东北林业大学林木遗传育种与生物技术教育部重点实验室,哈尔滨 150040) (2.牡丹江师范学院,牡丹江 157012)
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2008-05-20 发布日期:2008-05-20
  • 通讯作者: 杨传平
  • 基金资助:
     

Construction and EST Analysis of Suppression Subtractive Hybridization Library of Betula platyphylla Female Inflorescence

WANG Chao;YANG Chuan-Ping*;WEI Ji-Cheng;JIANG Jing   

  1. (1.Key Laboratory of Forest Tree Genetic Improvement and Biotechnology,Ministry of Education Northeast Forestry University,Harbin 150040) (2.Mudanjiang Teachers College,Mudanjiang 157012)
  • Received:1900-01-01 Revised:1900-01-01 Online:2008-05-20 Published:2008-05-20
  • Contact: YANG Chuan-Ping
  • Supported by:
     

摘要: 为研究白桦雌花序发育过程中特异基因的表达,以白桦雌花序样品为tester,雄花序样品为driver,利用SMART策略构建了白桦雌花序抑制性消减(SSH)文库。构建SSH文库的重组率为72%,插入片段的平均长度为400 bp左右。随机挑选文库克隆测序,获得150条EST序列,这些序列被GenBank的dbEST数据库收录,收录号为EE284580-EE284681,EE595316-EE595363。通过BlastX对EST进行功能注释,并对其中同源性较高的111条EST按功能进行分类,EST功能涉及了代谢、细胞防御、转录调节、能量代谢及信号传导等途径。发现了多个已知的控制花发育相关的EST,它们占已知功能EST的21%其功能涉及到调控花序形成和花分化、调控花粉与柱头亲和性以及调控花粉管发育等,包括MADS-box、S-locus F-box等基因。这些EST的获得为了解白桦花期基因表达,白桦花发育相关基因克隆和功能解析奠定了基础。

关键词: 白桦, 发育, 表达序列标签, 雌花, 抑制性消减文库

Abstract: To investigate the differentially expressed genes during the development of Betula platyphylla female inflorescence, a suppression subtractive hybridization (SSH) library was constructed by using the cDNAs from the female and male inflorescences, respectively, as the tester and the driver. The recombination of the SSH library was 72%, with about 400 bp of average insert length. The clones from SSH library were selected randomly for sequencing. Totally, 150 ESTs were obtained, with the GenBank accession number EE284580-EE284681, EE595316-EE595363. The ESTs were functional annotated by BlastX. Among them, 111 ESTs significantly matched with Nr database, and were further classified into functional groups including metablism, cell rescue, defence, transcription, energy and signal transduction, etc.. The genes related in flower development were found in our EST collection, which account for 21% of known function ESTs. These genes including MADS-box, S-locus F-box, etc., and their function involved in inflorescence forming and differentiation regulation, pollen-stigma interactions control and pollen tube development, etc. The obtaining of these EST will provide a reference for analysis of gene expression in the inflorescence of B.platyphylla, and also will make a base for cloning genes related with developments of follower and gene functional analysis in follower development.

Key words: Betula platyphylla Suk, development, expressed sequence tag, female flower, suppression subtractive hybridization library

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