CAJ | 학술논문

为分离辣椒细胞质雄性不育基因,并阐明其不育机理,本研究以辣椒细胞质雄性不育系21A及保持系21B的线粒体DNA为材料进行SRAP分析。结果显示,在21A中获得3条多态性条带。对多态性条带回收、克隆和测序分析后发现,这3个片断(分别命名为CMS721、CMS394、CMS285)在GenBank中都与能量代谢途径有关。针对序列特征设计SCAR引物,对21A和21B的基因组DNA进行扩增验证,3对引物只在21A中扩增出目的条带,表明已成功地将SRAP标记转化为SCAR标记。利用荧光定量PCR技术对CMS721、CMS394和CMS285基因在21A不同组织(根、茎、叶和花)中的表达情况进行了分析。结果表明：3个基因只在不育系中表达,且在不同的组织中均有表达,但表达量差异很大,其中基因CMS721表达量在中花蕾中迅速下降,推测该基因控制辣椒花药中与能量代谢相关的蛋白质的表达,从而引起小孢子发育障碍,最终导致雄性不育。
위분리랄초세포질웅성불육기인,병천명기불육궤리,본연구이랄초세포질웅성불육계21A급보지계21B적선립체DNA위재료진행SRAP분석。결과현시,재21A중획득3조다태성조대。대다태성조대회수、극륭화측서분석후발현,저3개편단(분별명명위CMS721、CMS394、CMS285)재GenBank중도여능량대사도경유관。침대서렬특정설계SCAR인물,대21A화21B적기인조DNA진행확증험증,3대인물지재21A중확증출목적조대,표명이성공지장SRAP표기전화위SCAR표기。이용형광정량PCR기술대CMS721、CMS394화CMS285기인재21A불동조직(근、경、협화화)중적표체정황진행료분석。결과표명：3개기인지재불육계중표체,차재불동적조직중균유표체,단표체량차이흔대,기중기인CMS721표체량재중화뢰중신속하강,추측해기인공제랄초화약중여능량대사상관적단백질적표체,종이인기소포자발육장애,최종도치웅성불육。
In order to isolate cytoplasmic mal sterility related genes and clarify the mechanism in pepper, sequence-related amplified polymorphism (SRAP) was used to analyze mitochondrial DNA of cytoplasmic male sterility line 21A and its maintainer line 21B of pepper. Three polymorphism fragments detected in 21A were named CMS721、CMS394 and CMS285 in GenBank were related to energy metabolism. According to the sequences, three specific primers designed amplified tar-get DNA in the 21A line, indicative of transformation of SRAP marker to SCAR marker. Expression level of the genes ( CMS721、CMS394 and CMS285 ) in different tissues ( root, leaf, stem and flower) analyzed by quantitative real-time poly-merase chain reaction (qRT-PCR) revealed that the three genes were only expressed in tissues of 21A. The expression lev-el of CMS721 gene decreased rapidly in the middle flower bud stage, suggesting that CMS721 might regulate the expression of energy metabolism related protein in anther, causing microspore abortion and cytoplasmic male sterility.