Differential Expression Analysis of Genic Male Sterile-fertile Line in Pepper (Capsicum annuum L.) by cDNA-AFLP | 利用cDNA-AFLP技术研究辣椒核雄性不育两用系的基因差异表达

中国人. 利用辣椒雄性不育生产杂交种取得了重大进展，但有关雄性不育的分子机制仍不清晰。本研究利用cDNA-AFLP技术，比较分析了辣椒(Capsicum annuum L.)核雄性不育两用系中不育植株与可育植株花药发育过程中基因表达的差异。用1024对选择性扩增引物进行筛选，共获得了168个转录衍生片段(transcript-derived fragment, TDFs)。通过BLAST比对，多数TDFs未找到对应的同源序列，可能代表了新基因；48个TDFs在数据库中找到了同源序列(GenBank登录号: JK993564~JK993611)，对这些序列进行功能分类，差异表达的基因涉及基础代谢(25%)、胁迫反应(4.2%)、转录调控(14.6%)、信号转导(6.3%)、植物发育(10.4%)等多方面的功能。对其中24条TDFs进行半定量RT-PCR检测，分析其在可育株和不育株不同组织及花药发育不同时期的表达模式，结果表明11条TDFs有时空表达特异性，推测和雄性不育相关。其中TDF14和TDF21为花药特异表达，TDF39只在可育花药及可育株和不育株的萼片中特异表达。以上研究为揭示辣椒核不育的分子调控机理提供理论依据，同时为后续的分子辅助育种和育性相关基因的克隆提供基础资料。

英语. Hybrid production using male sterile in pepper has been made great progress. However, the molecular mechanism of male sterile is still not clear. Using anthers as material, differential expression of genic male sterile-fertile line in pepper (Capsicum annuum L.) was analyzed by cDNA-AFLP. One hundred and sixty-eight transcript-derived fragments (TDFs) were obtained. Most of the TDFs were not found homologous sequence in GenBank, probably standing for new genes. Forty-eighty TDFs were found homologous sequence(GenBankAccession No.: JK993564~JK993611). These genes were involved in primary metabolism (25%), signal transduction (6.3%), plant development (10.4%), stress response (4.2%), transcription (14.6%), etc.. Twenty-four TDFs were chosen for semi-quantitative RT-PCR to detect the expression pattern in various tissues and different development stages of anther in fertile and sterile plants. The results showed that 11 TDFs displayed expression specificity in time and space and might be male sterile related genes. TDF14 and TDF21 were anther specifically expressed genes and TDF39 specifically expressed in fertile anthers and sepals of fertile and sterile plants. These findings can provide a basic data for molecular mechanism research of genic male sterile, subsequent marker assisted selection and cloning of fertility-related genes in pepper.