EST-SSR 마커 적용을 통한 수박 F1 품종 순도 검정 = Application of EST-SSR Marker for Purity Test of Watermelon F1 Cultivars

다국어 초록 (Multilingual Abstract)

This study was conducted to develop a set of EST-SSR marker for the purity test of commercial F1 hybrid cultivars in the watermelon. A total of 353 EST-SSR were selected and tested on seven F1 cultivars and their 11 parental lines achieved from NH Seeds Inc., Korea. Among tested 96 primer sets, WMU0056 for 'Orange', WMU0400 for 'Heukbo', WMU0056 and WMU0400 for 'Sindong', and WMU0056 and WMU0400 for 'Serona' revealed polymorphisms between the parental lines and heterozygosity from these F1 cultivers. Of 122 primer sets tested for 'Haedong', WMU0056, WMU0400, WMU0580, WMU1211, WMU4136, and WMU448 showed polymorphisms that were appropriate for the F1 purity test. WMU0056 and WMU0400 can be useful for 'Haedong', as well. Relatively low polymorphisms between parental lines were detected for 'Kulnara'(5%) and 'Hwangpea'(2%), and therefore, all 353 primer sets were tested on these cultivars. As the result, WMU5339 and WMU7003 were found to be useful for the F1 purity test in 'Kulnara' and 'Hwangpea', respectively. Using these EST-SSR markers developed by ICuGI, hybridity of the seeds for four F1 cultivars produced from farmers was evaluated, and levels of the F1 purity higher than 97.5% was observed from all seed populations. Our results indicated that the watermelon EST-SSR marker information posted in ICuGI could be utilized for developing codomiant and locus-specific markers that are highly effective for the F1 purity test.

국문 초록 (Abstract)

본 연구는 ICuGI Database를 활용하여 수박 상용 F1 품종의 순도검정용 EST-SSR 마커를 개발하기 위해 수행되었다. 총 353개 EST-SSR primer set을 선발하여 (주)NH종묘의 수박 F1 품종 7종과 각 품종의 양친 11종에 대해 검정하였다. 이중 1차 테스트한 96개 primer set 중, ‘오렌지’는 primer WMU0056, ‘흑보’는 WMU0400, ‘신동’은 WMU0056와 WMU0400, ‘새로나’는 WMU0056, WMU0400, WMU0529에 대해 각 품종의 양친들이 다형성이 보였고 F1 개체에서는 이형접합의 유전자형을 보였다. ‘해동’ F1의 순도검정용 마커를 찾기 위해 추가적인 122개의 primer set에 대해 PCR을 수행한 결과, WMU0056, WMU0400, WMU0580, WMU1211, WMU4136, WMU448이 순도검정에 적합한 것으로 나타나, WMU0056와 WMU0400이 ‘해동’에서도 유용할 수 있었다. ‘꿀나라’와 ‘황피’의 경우에는 공시된 타 품종들에 비해 양친간 다형성율이 각각 5%와 2%로 매우 낮아 모든 353개 primer set을 테스트하였으며, 그 결과 ‘꿀나라’는 WMU5339, ‘황피’는 WMU7003이 순도검정 마커로 적합한 것으로 확인되었다. 개발된 마커를 이용하여 실제 농가채종된 4개의 F1 품종의 순도를 검정한 결과, 모두 97.5% 이상의 순도로 확인되었다. 이와 같이 ICuGI Database에 공시된 수박 EST-SSR 마커는 공우성의 유전자 특이적 마커로서 F1 순도검정에 효과적으로 사용될 수 있었다.

참고문헌 (Reference)

1 MFAFF, "Ministry for Food, Agriculture, Forestry and Fisheries. Statistics information"

2 KSA, "Korea Seed Association" Statistics information

3 Kong, Q, "Identification and development of polymorphic EST-SSR markers by sequence alignment in pepper, Capsicum annuum" e59-e61, 2012

4 Staub, J. E, "Genetic markers, map construction, and their application in plant breeding" 31 : 729-741, 1996

5 Hwang, J. H, "Genetic diversity in watermelon cultivars and related species based on AFLPs and EST-SSRs" 39 : 285-292, 2011

6 Levi, A, "Genetic diversity among watermelon (Citrullus lanatus and Citrullus colocynthis) accessions" 48 : 559-566, 2001

7 Hwang, J. H, "Functional characterization of watermelon (Citrullus lanatus L.) EST-SSR by gel electrophoresis and high resolution melting analysis" 130 : 715-724, 2011

8 Levi, A, "EST-PCR markers representing watermelon fruit genes are polymorphic among watermelon heirloom cultivars sharing a narrow genetic base" 7 : 16-32, 2008

9 Ritschel, P. S, "Development of microsatellite markers from an enriched genomic library for genetic analysis of melon (Cucumis melo L.)" 4 : 1471-2229, 2004

10 Guerra-Sanz, J. M, "Citrullus simple sequence repeats markers from sequence databases" 2 : 223-225, 2002

11 Hyo Guen Park, "Application of RAPD and SCAR Markers for Purity Testing of F1 Hybrid Seed in Chili Pepper (Capsicum annuum)" 한국분자세포생물학회 18 (18): 295-299, 2004

12 Levi, A, "An extended linkage map for watermelon based on SRAP, AFLP, SSR, ISSR, and RAPD markers" 131 : 393-402, 2006