茄子雜交一代種子純度鑑定技術開發

Development of technology for purity identification of eggplant hybrids seeds

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

《臺南區農業改良場研究彙報》 78期 (2021/12) Pp. 22-31

摘要

隨著分子技術的快術發展，種子純度檢定除了以往傳統的生育特性檢測方法（grow out test, GOT），亦可選擇使用更快速之分子檢測技術進行，本試驗利用限制酶位點標定定序方法（restriction-site associated DNA sequencing, RAD-seq），定序了17個茄子品種，經分析獲得了2,972個在參試品種間可靠的單一核苷酸多型性位點（single nucleotide polymorphisms, SNPs）。並利用30個PIC值大於0.4的SNPs進行kompetitive allele specific PCR（KASP）分子標誌開發，藉由即時聚合酶連鎖反應器進行終點螢光量分析。試驗結果顯示，20組KASP分子標誌可用於茄子雜交一代種子純度鑑定與品系純度檢定工作。綜合上述結果，此套技術可協助國內茄子種苗業者簡易且快速的進行茄子雜交種子純度檢測，或是協助育種工作的進行，進而提升茄子雜交種子品質及加速育種速度。現有技術：目前種苗產業多數以傳統的生育特性檢測法進行雜交種子的純度鑑定，所需時間較長，也需較大栽植空間導致成本較高。創新內容：本研究透過KASP分子標誌建立茄子雜交品種的純度檢測技術，並將其應用於品系純化作業。對產業影響：協助茄子種苗業者以簡易且快速的方法進行茄子雜交種子純度檢測，或是應用於育種的純度檢定工作。

關鍵字

次世代定序 ； 限制酶位點標定定序方法 ； 雜交種子純度檢測

並列摘要

In addition to the traditional grow out test (GOT), seed purity verification can also use rapid molecular detection technology. In this experiment, the restriction-site associated DNA sequencing (RAD-seq) technique was used to sequence 17 eggplant varieties, and 2,972 reliable single nucleotide polymorphisms (SNPs) were obtained from analysis. We used 30 SNPs with a PIC value greater than 0.4 to develop kompetitive allele specific PCR (KASP) and analyzed the end-point fluorescence on real-time PCR detection system. Our results showed that 20 KASP markers can be used for the purity identification and line purification of eggplant hybrid seeds. Based on the results, these KASP markers can help domestic seedling companies detect the purity of eggplant hybrid seeds, improve quality, and accelerate the breeding process in an easier and quicker way. What is already known on this subject? Currently, most seedling industries use grow out test (GOT) to identify the purity of hybrid seeds. However, it takes longer time and much space cost. What are the new findings? In this study, we established a purity detection technology through KASP markers and applied them to purity identification of eggplant hybrid seeds and line purification. What is the expected impact on this field? To assist breeding companies in controlling the purity of eggplant hybrid seeds and line purification.

並列關鍵字

Next Generation Sequencing ； Restriction-site associated DNA sequencing ； Detection of hybrid seeds purity