A novel strong candidate gene CA00g82510 for the male fertility restoration locus CaRf032 in Capsicum annuum was identified by genome re-sequencing and recombination analysis. A single dominant locus (CaRf032) for fertility restoration of cytoplasmic male sterility was identified in the strong restorer inbred line IVF2014032 of chili pepper (Capsicum annuum L.). CaRf032 was localized within an 8.81-Mb candidate intervals on chromosome 6 using bulked segregant analysis based on high-throughput sequencing data. Subsequently, the candidate interval was genetically mapped and defined to a 249.41-kb region using an F2 population of 441 individuals generated by crossing the male-sterile line 77013A and the restorer line IVF2014032. To fine map CaRf032, eight newly developed KASP markers were used to genotype 23 recombinants screened from a larger F2 population of 2877 individuals. The CaRf032 locus was localized to a 148.05-kb region between the KASP markers S1402 and S1354, which was predicted to contain 22 open reading frames (ORFs). One ORF with an incomplete sequence was predicted to contain a PPR motif, and its physical position overlapped with the Rf candidate gene CaPPR6_46. The PPR ORF sequence before the gap showed 100% identity with the CA00g82510 locus of the CM334 reference genome. CA00g82510 encodes a protein of 583 amino acids, containing 14 PPR motifs, and shows significantly differential expression between the flower buds of the maintainer line 77013 and the restorer line IVF2014032. These results indicated that CA00g82510 is a strong candidate gene for CaRf032. Five KASP markers, which detected single-nucleotide polymorphisms in CA00g82510 of 77013 and IVF2014032, co-segregated with CaRf032 and showed 64.4% successful genotyping of 38 maintainer and 63 restorer lines.

中文翻译：

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辣椒雄性生育力恢复基因CaRf032的精细定位。

通过基因组重测序和重组分析，确定了辣椒雄性生育力恢复位点CaRf032的新强候选基因CA00g82510。在辣椒 (Capsicum annuum L.) 的强恢复系自交系 IVF2014032 中鉴定了一个用于恢复细胞质雄性不育生育力的显性基因座 (CaRf032)。使用基于高通量测序数据的批量分离分析，将 CaRf032 定位在 6 号染色体上 8.81-Mb 的候选区间内。随后，使用通过雄性不育系 77013A 和恢复系 IVF2014032 杂交产生的 441 个人的 F2 群体对候选区间进行遗传映射并定义为 249.41-kb 区域。精细映射CaRf032，8 个新开发的 KASP 标记用于对从 2877 个个体的较大 F2 群体中筛选的 23 个重组体进行基因分型。CaRf032 基因座定位于 KASP 标记 S1402 和 S1354 之间的 148.05-kb 区域，预计该区域包含 22 个开放阅读框 (ORF)。一个具有不完整序列的 ORF 被预测包含一个 PPR 基序，并且其物理位置与 Rf 候选基因 CaPPR6_46 重叠。缺口前的 PPR ORF 序列与 CM334 参考基因组的 CA00g82510 基因座显示出 100% 的同一性。CA00g82510编码583个氨基酸的蛋白质，包含14个PPR基序，在维持系77013和恢复系IVF2014032的花蕾之间表现出显着差异表达。这些结果表明CA00g82510是CaRf032的强候选基因。