高粱雄不稔細胞質SCAR分子標誌之開發

Development of a SCAR Marker for Male Sterile Cytoplasm in Sorghum

龔美玲(Mei-Ling Kung) ； 詹詩褕(Shih-Yu Jhan) ； 張惠如(Hui-Ju Chang)

《植物種苗》 19卷2期 (2017/06) Pp. 59-72

摘要

高粱普遍利用雄不稔特性生產一代雜交品種，但沒有相關的連鎖標誌可應用於雄不稔細胞質的鑑別，因此本研究以本土高粱雜交品種‘臺中5號’及其親本品系雄不稔親‘80A’與維持親‘80B’為材料，設計與雄不稔有關的粒線體atp6、coxII、atp1-1及atp9基因專一性引子對進行序列多型性分析，試驗結果發現‘80A’與‘80B’在atp1-1之3’端具有多型性，進而設計SCAR標誌（SB-atp1-1_indel_F2/R2）可在雄不稔親‘80A’擴增401 bp大小的條帶，在維持親‘80B’則可擴增701 bp的條帶。比較兩者PCR產物序列差異，發現‘80A’品系在atp1-1 3'端UTR缺失（deletion）的部分序列中含有短片段的重複序列。綜合上述本研究獲得之分子標誌可區分近似同源系的‘80A’與‘80B’，並可應用於確認高粱育種之細胞質稔性遺傳背景及快速鑑別品系之種子純度。

關鍵字

高粱 ； 雄不稔 ； 分子標誌

並列摘要

Male sterile is widely used in hybrid seed production in sorghum. Domestic sorghum hybrid variety 'Taichung No. 5' and its parental lines were used as materials in this study. In order to quickly identify the cytoplasmic genetic background of fertility, specific markers were designed for CMS-associated mitochondrial genes, including atp6, coxII, atp1-1 and atp9. PCR products derived from male sterile line '80A', the sterile parental line of 'Taichung No. 5', and its maintain line '80B' were polymorphic at the 3' end of atp1-1. The SCAR marker (SB-atp1-1_indel_F2/R2) amplified the 401 bp band in line '80A' and the 701 bp band in line '80B', respectively. DNA sequencing of PCR products showed that the repeat sequences existed in the 3'UTR of atp1-1 and the deletion region of '80A' line. These results showed that the SCAR marker could distinguish the near-isogenic lines, '80A' and '80B', and be applied in rapid identification for cytoplasm types in breeding program and purity inspection of sorghum.

並列關鍵字

sorghum ； male sterile ； molecular marker