棉花花药全生育期均一化全长cDNA文库的构建和分析

doi:10.3864/j.issn.0578-1752.2013.18.001

摘要/Abstract

摘要： 【目的】深入了解棉花花药发育的分子机制，并为其不育机理的研究提供参考序列。【方法】构建棉花花药全生育期的均一化全长cDNA文库并测序获得9 896条高质量的EST，利用生物信息学和荧光定量相结合法对所得数据进行系统分析。【结果】该cDNA文库库容为1.2×107，插入片段分布在1—3 kb。将所测得的高质量EST进行拼接得到6 643条unigenes，平均长度为779 bp。经过序列比对注释和生物信息学分析，得到一系列在棉花花药发育过程中起重要作用的基因，并获得28条参与花粉壁合成的基因。荧光定量结果表明，这些基因均在花药发育前期优势表达，尤其是在四分体时期表达量最高。【结论】获得大量的棉花花药表达基因，并分析得到一些调节棉花花药发育的重要途径以及参与棉花花粉壁合成相关基因。

关键词: 陆地棉 , cDNA文库 , 花药 , 花粉壁

Abstract: 【Objective】The objective of this study is to understand the anther development and make a firm foundation for sterile mechanism in upland cotton (Gossypium hirsutum L.). 【Method】A normalized full-length cDNA library from anthers including the full development stage was firstly constructed and 9 896 high-quality ESTs were sequenced in this study. The bioinformatics analysis and qRT-PCR were performed to deep analyze this cDNA library. 【Result】The storage capacity of this cDNA library is 1.2×107 and the average length of the inserted cDNA fragment was dispersed between 1 kb and 3 kb. These high-quality ESTs were further assembled into 6 643 unigenes with 779 bp in average length. Through bioinformatics analysis of these unigenes, many important biological processes were obtained during the anther development and 28 genes related with the biosynthesis of pollen wall were identified during anther development. The qRT-PCR was performed to check the expression pattern of these pollen wall biosynthesis related genes and all of them were high-expressed in early anther development especially in tetrad pollen period. 【Conclution】In this study, lots of anther expressed genes were firstly sequenced in upland cotton. Many important processes regulating anther development were obtained and many genes related with pollen wall biosynthesis were obtained.

Key words: Gossypium hirsutum L. , cDNA library , anther , pollen wall

马建辉., 刘记., 范术丽, 宋美珍, 庞朝友, 魏恒玲, 喻树迅. 棉花花药全生育期均一化全长cDNA文库的构建和分析[J]. 中国农业科学, 2013, 46(18): 3743-3749.

MA Jian-Hui-., LIU Ji-., FAN Shu-Li, SONG Mei-Zhen, PANG Chao-You, WEI Heng-Ling, YU Shu-Xun. Construction and Analysis of Normalized Full-Length cDNA Library of Upland Cotton Anthers[J]. Scientia Agricultura Sinica, 2013, 46(18): 3743-3749.

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