茄子温敏单性结实抑制差减文库的构建与分析

doi:10.3864/j.issn.0578-1752.2011.16.009

摘要/Abstract

摘要： 【目的】构建茄子单性结实差减文库，分离获得茄子单性结实相关基因，研究茄子单性结实形成的分子机制。【方法】以茄子单性结实品系D-10低温条件下无籽的子房和果实，以及适温条件下有籽的子房和果实为材料，采用抑制差减杂交（SSH）技术构建正向和反向差减文库。对阳性克隆测序、BLAST比对分析、GO（gene ontology）功能注释及分类和荧光定量PCR分析。【结果】正反向文库共获得了2 347个阳性克隆；测序、序列拼接后，共获得1 248个高质量的unigenes；将其与非冗余数据库BLAST比对后，有1 109个unigenes找到同源序列，139个unigenes无同源序列为新基因。1 248个unigenes中527个在GO数据库中有功能注释，其中细胞组分、分子功能和生物过程的unigenes分别为206、445和361条，分析表明，茄子单性结实果实的差异主要发生在细胞内部，单性结实性状的表达可能受到一些因子和激酶的调控。【结论】成功构建了茄子单性结实不同时期果实的抑制差减文库，获得3个可能与茄子单性结实相关的unigenes，包括MADS-box基因、雌蕊伸展相关蛋白和果实膨大相关基因。

关键词: 茄子, 单性结实, 抑制差减杂交, EST

Abstract: 【Objective】The aim of this experiment is to investigate the parthenocarpy-related genes in eggplant, and study the parthenocarpic molecular mechanism. 【Method】One forward and one reverse cDNA library were constructed by suppression subtractive hybridization (SSH). All the materials are from the same facultative parthenocapic line D-10, including the parhtenocarpic ovaries and fruits at low temperature, and the unparthenocarpic ovaries and fruits at optimal temperature. The positive clones of the libraries were sequenced randomly, analysed by BLAST， classified by GO and analysed by real-time quantitative PCR. 【Result】A total of 2 347 positive clones in the forward and reverse libraries were obtained. After sequenced and assembled, 1 248 high-quality unigenes were gotten. Blast analysis was made with no redundant database, 1 109 of these unigenes were homologous to known genes, and 139 could be new genes. In Gene Ontology database, 527 unigenes were assigned functional description, and 206, 445, 361 ESTs were, respectively, involved in cell component, molecular function and biological process. According to classification results, the difference between parthenocarpy and unparthenocarpy almost existed in intracellular region. Parthenocarpy may be regulated by some factors and kinases, and most of the differentially expressed genes were involved in basic biological processes.【Conclusion】The SSH cDNA libraries were successfully constructed. Three parthenocarpic related unigenes, including MADS-box gene, pistil extensin-like protein and fruit-ripening gene, were obtained and the possible mechanism was analyzed.

Key words: eggplant, parthenocarpy, SSH, EST

张映, 陈钰辉, 张振贤, 方智远, 连勇, 刘富中. 茄子温敏单性结实抑制差减文库的构建与分析[J]. 中国农业科学, 2011, 44(16): 3368-3376.

ZHANG Ying, CHEN Yu-Hui, ZHANG Zhen-Xian, FANG Zhi-Yuan, LIAN Yong, LIU Fu-Zhong. Construction and Analysis of Eggplant Thermo-Sensitive Parthenocarpy SSH cDNA Library[J]. Scientia Agricultura Sinica, 2011, 44(16): 3368-3376.

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