利用SNP分子標誌建立胡瓜雜交種子純度檢測技術

Establishment of SNP detection technology to examine the purity of cucumber hybrids

王聖善(Wang, S. S.) ； 朱詠筑(Y. C. Chu) ； 蔡佩樺(P. H. Tsai) ； 楊藹華(A. H. Yang)

《臺南區農業改良場研究彙報》 74期 (2019/12) Pp. 27-35

摘要

種子純度鑑定是雜交種子生產過程中品質管控的重要工作，為了協助國內種苗業者檢測胡瓜一代雜交品種的雜交成功率，我們藉由雙限制酶位點標定定序方法（double digest restriction-site Associated DNA sequencing, ddRAD-seq）定序了62個胡瓜品種，經分析獲得了2,121個於參試品種間可靠的單一核苷酸多型性位點（single nucleotide polymorphisms, SNPs）。為了建立可運用於高通量的基因型分析方法，我們選取了均勻分布於胡瓜7條染色體的31個PIC（polymorphism information content）值大於0.35的SNPs，將其設計成KASP（kompetitive allele specific PCR）分子標誌。實驗結果顯示，總共有20個KASP分子標誌可準確的區分參試樣品基因型。此套分子標誌經65個國內一代雜交的胡瓜品種測試發現，每個品種平均具有7.6可用來執行胡瓜雜交成功率檢測的分子標誌。綜合以上結果，我們認為此套分子標誌可快速且精確執行胡瓜雜交種子的純度鑑定，用以提升國內雜交種子的生產品質。現有技術：目前種苗產業多數以生育特性的檢測方法執行雜交種子的純度鑑定。創新內容：本研究透過KASP分子標誌建立胡瓜雜交品種的純度鑑別技術。對產業影響：此技術可提供種苗業者快速且精準的檢測胡瓜雜交種子純度，提升胡瓜種子品質。

關鍵字

次世代定序 ； 雙限制酶位點標定定序方法

並列摘要

Purity testing is one of the key tasks in quality control of hybrid seed production. This study is attempted to establish the single nucleotide polymorphisms (SNPs) systems for assessment of cucumber hybrid seed purity, 62 cucumber varieties were sequenced by double digest restriction-site associated DNA sequencing (ddRAD-seq) methods. After analyzing, 2,121 reliable SNPs were detected. A total of 31 SNPs with high polymorphism information content (PIC) value (＞ 0.35), and evenly located on 7 cucumber chromosome were designed as kompetitive allele specific PCR (KASP) markers. Eventually, 20 out of the 31 KASP markers showed high call rate. These KASP markers could accurately and rapidly assess the purity of hybrid seeds, and could be used to improve the hybrid seed quality. What is already known on this subject? No purity assessment system of cucumber hybrids using allele-specific PCR platform has been developed in Taiwan. What are the new findings? In this study, we established a purity assessment system of cucumber hybrids based on 20 KASP markers. What is the expected impact on this field? With these markers, breeding companies could accurately and rapidly control the quality of cucumber hybrids

並列關鍵字

Next Generation Sequencing ； Double digest Restriction-site Associated DNA sequencing