辣椒胞质雄性不育恢复基因CAPS标记的PARMS转换及其应用

CAPS Marker of Cytoplasmic Male Sterility Restorer-of-fertility Gene in Pepper(Capsicum annuum)Converted to PARMS Marker and Its Application

Co1Mod1-CAPS是一个可广泛应用于筛选辣椒胞质雄性不育恢复源的CAPS分子标记。为了将其转化为可高通量检测的分子标记以提高选择效率,本研究以Co1Mod1-CAPS标记扩增10份不同类型的辣椒三系材料,分析其扩增产物在不育系、保持系和恢复系材料间的序列差异,从而基于序列差异将其转化为PARMS高通量检测标记并对321份辣椒种质材料进行恢复源筛选。结果表明,在Co1Mod1-CAPS标记扩增产物第601个碱基处存在一个T→C碱基的变异,即不育系和保持系材料在该处的碱基均为T,而恢复系材料在该处的碱基为C,后基于该碱基变异成功将Co1Mod1-CAPS转化为高通量标记Co1Mod1-PARMS,并在321份种质材料中筛选出146份资源携带纯合恢复基因RfRf,其中小果形且辣味重的辣椒类型携带RfRf基因型的个体占比高,指形椒的占比最高,为59.5%,樱桃椒占比次之,为56.5%,甜椒类型辣椒(灯笼椒)携带RfRf基因型的个体占比最低,为8.3%。该研究结果与前人研究较为一致,表明该标记有效。因此,Co1Mod1-PARMS高通量检测标记可应用于辣椒胞质雄性不育恢复系的分子标记辅助选育,从而促使胞质雄性不育恢复系选育效率的提升。

Co1Mod1-CAPS is a molecular marker that could be widely used to screen for sources carrying restorer-of-fertility gene associated with cytoplasmic male sterility in pepper.In order to enhance its utility as a high-throughput detectable molecular marker and improve selection efficiency,10 three-line(sterile line,maintainer line and restorer line)materials belonging to different types of pepper were amplified using Co1Mod1-CAPS marker,and the sequence differences of the amplification products amongthem were analyzed.Based on these sequence differences,we converted Co1Mod1-CAPS marker into PARMS marker,which allows for high throughput detection.The PARMS marker was subsequently employed to screen 321 for materials carrying the restorer-of-fertility gene across 321 pepper germplasm sources.The results revealed a T→C base variation at the 601st position of the Co1Mod1-CAPS marker amplification product,with T in sterile and maintainer lines and C found in restorer lines.Based on this base variation,we successfully converted Co1Mod1-CAPS into the high-throughput marker Co1Mod1-PARMS,and screened out 146 resources carrying the homozygous restorer-of-fertility gene RfRf among the 321 germplasm materials.Among them,the small-fruit-shaped and spicy pepper had a higher proportion of individuals carrying the RfRf genotype.The short finger-shaped pepper had the highest proportion at 59.5%,followed by cherry pepper at 56.5%,while the sweet pepper(lantern pepper)had the lowest proportion at 8.3%.The results of this study are consistent with previous research,confirming the effectiveness of Co1Mod1-PARMS.Therefore,this high-throughput marker can be used for molecular marker-assisted breeding of cytoplasmic male sterile restorer lines in peppers,thereby improving the breeding efficiency of cytoplasmic male sterile restorer lines.