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以辣椒细胞质雄性不育系21A及其相应保持系21B为试材,根据辣椒细胞质雄性不育相关基因orf456序列设计特异引物,对21A及21B的基因组DNA进行特异性扩增,获得了不育系特有的目的条带,克隆测序表明其大小为292bp,命名为CMS292。采用QRT-PCR方法,研究了CMS292分别在蕾期、盛花期对根、茎、叶和花蕾中的表达水平的影响,以期探索雄性不育相关基因的表达机制。结果表明:该基因只在不育系中表达,并且在所检测的组织中都有表达;蕾期根和花蕾中的表达量明显高于茎和叶,盛花期根和叶中的表达量高于茎和花蕾;从蕾期到盛花期根、茎和花蕾中的表达量呈下降趋势,而叶中的表达量呈上升趋势,推测CMS292控制辣椒花药中蛋白的表达,引起小孢子败育,进而导致雄性不育。
Based on the orf456 sequence of pepper cytoplasmic male sterility (CMS) -based primers, the genomic DNA of 21A and 21B was amplified by specific PCR using the CMS line 21A and its maintainer line 21B. The specific purpose of the band, cloning and sequencing showed that the size of 292bp, named CMS292. The effect of CMS292 on the expression level of roots, stems, leaves and buds at budding and full flowering stages was investigated by QRT-PCR. The purpose of this study was to explore the mechanism of CMS292 expression. The results showed that the gene was expressed only in CMS lines and expressed in all tested tissues. The expression level in buds and buds was significantly higher than that in stems and leaves, In stems and flower buds, the expression levels in roots, stems and buds from bud stage to full flowering stage decreased while the expression levels in leaves showed an upward trend. It was speculated that CMS292 could control the expression of proteins in pepper anthers, causing microspore abortion, Which in turn leads to male infertility.