辣椒雄不稔系統分子標誌之研發

Abstract

番椒 (Capsicam annuum L.) 為茄科 (Solanaceae) 番椒屬，一般分為辣椒及甜椒兩群。現今商業生產上常採用雄不稔親 (S/rfrf)、維持親 (N/rfrf)、與恢復親 (N/RfRf) 大量生產雜交種子，並稱此系統為細胞質雄不稔系統。為建立可鑑別細胞質雄不稔系統的辣椒分子標誌，本研究利用 141 個辣椒細胞質雄不稔系統親本為材料，進行與細胞質特性及稔性恢復基因有關的分子標誌研究。與細胞質特性有關的分子標誌研究，對 atp6-2 序列進行分析，在 141 個品系當中得到四種 haplotype 的表現，其中具雄不稔細胞質的雄不稔親本均為第一種 haplotype 的表現，且與 ψatp6-2 前段序列 SNP 表現一致，並設計區別 atp6-2 基因變異的多引子系統區分四種 haplotype。與稔性恢復基因有關的分子標誌研究，利用 CRF 及 AFRF8CAPS 兩組分子標誌進行 141 個品系的普遍性檢測。CRF 分子標誌普遍性為 93％，AFRF8CAPS 分子標誌普遍性為 3.5％，並對 CRF 與 AFRF8CAPS 分子標誌顯、隱性表現的條帶進行序列分析，得到序列間的 SNP 差異位置。

Pepper (Capsicum annuum L.) has two groups：chili pepper and sweet pepper, belongs to the Solanaceae family. Cytoplasmic-genetic male sterility (CGMS), commonly referred as cytoplasmic male sterility (CMS) system has been utilized for F1 hybrid seeds production, is composed of the sterile line (S/rfrf), maintainer line (N/rfrf) and restorer line (N/RfRf). In this research, we use 141 CMS system parents to develop molecular markers associate with the sterile cytoplasm and the nuclear fertility restorer gene (Rf). Four types of sequence composition were found for the ribosomal atp6-2 gene, of which the Type I sequence is always associated with the truncated pseudo-gene ψatp6-2 as well as the male sterility characteristic of cytoplasm. A multi-primer system has been designed to identify the Type I atp6-2 sequence and the ψatp6-2 sequence. Two markers, namely CRF and AFRF8CAPS, from published literatures, were used to exam the association between them and the Rf gene. For the 141 CMS system parents studied, CRF is associated with Rf gene in 93% of the lines, whereas AFRF8CAPS is associated with Rf gene in 3.5% of the lines. DNA Sequences of both the dominant and recessive alleles of these two molecular markers have been determined and the results reveal some SNPs that may be used for designing allele specific primers of PCR based co-dominant markers.