Development of Cleaved Amplified Polymorphic Sequence (CAPS) Marker for Selecting Powdery Mildew-Resistance Line in Strawberry (Fragaria × ananassa Duchesne)

Hee-Jeong Je; Jae-Wook Ahn; Hae-Suk Yoon; Min-Keun Kim; Jae-San Ryu; Kwang-Pyo Hong; Sang-Dae Lee; Young-Hoon Park

doi:10.7235/hort.2015.14133

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2015 Vol.33, Issue 5 Preview Page Next Page

Development of Cleaved Amplified Polymorphic Sequence (CAPS) Marker for Selecting Powdery Mildew-Resistance Line in Strawberry (Fragaria × ananassa Duchesne) 딸기 흰가루병 저항성 계통 선발을 위한 분자마커 개발

31 October 2015. pp. 722-729

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Abstract

Powdery mildew (PM) caused by Podosphaera aphanis is a major disease that can result in significant yield losses in strawberry (Fragaria × ananassa Duchesne). For preventing PM, pesticides are usually applied in strawberry. In this study, molecular markers were developed to increase breeding efficiency of PM-resistance cultivars by marker-assisted selection (MAS). An F2 population derived from a cross between PM-resistance ‘Seolhyang’ and PM-susceptibility ‘Akihime’ was evaluated for disease resistance to PM and RAPD (random amplification of polymorphic DNA)-BSA (bulked segregant analysis). Among 200 RAPD primers tested, OPE10 primer amplified a 311bp-band present in with 331bp. Sequence alignment performed for searching polymorphisms and six single nucleotide polymorphism (SNP) were found in amplified regions. To develop polymorphic marker for distinguishing between resistant and susceptible, RAPD was converted to cleaved amplified polymorphic sequence (CAPS) marker. Among restriction enzymes associated with six SNPs, Eae I (Y/GGCCR) was successfully digested to 231bp in susceptible. The results suggest that the selected CAPS marker could be used for increasing efficiency of selecting powdery mildew resistant strawberry in breeding system.

Keywords

bulked segregant analysis

marker-assisted selection

Podosphaera aphanis

딸기 흰가루병은 Podosphaera aphanis에 의해 발병되며 수확기에 가장 큰 피해를 주는 병으로 현재 유황, 농약으로 주로 방제 되고 있는 실정이다. 본 연구에서는 딸기 흰가루병 저항성 품종 육성을 위한 흰가루병 저항성 특이마커 개발로 내병성 육종효율을 높이고자 하였다. 흰가루병 저항성 계통 선발을 위한 분자마커를 개발하기 위해 아키히메×설향 집단을 대상으로 자가수분을 통해 후대 양성 후 병저항성을 검정하였다. 마커분석은 RAPD primer 200 세트 중 OPE10 331bp에서부터 흰가루병 저항성 특이 마커 선발하였다. 흰가루병 저항성 특이밴드만 선발하기 위하여 클로닝 후 유전자정보 분석하여 SP1F/R의 Primer를 제작하였다. 그러나 SP1F/R을 이용하여 PCR한 결과 저항성, 감수성간에 다형성이 확인되지 않아 염기서열을 정렬한 후 SNP, In/del의 다형성 유무를 확인한 결과 6개의 SNP를 확인하였다. 이들 PCR 산물을 해당 사이트와 연관된 제한효소로 절단한 결과 그 중 Eae I(Y/GGCCR)의 절단으로 231bp 위치에서 저항성과 감수성간의 다형성을 확인함으로써 흰가루병 저항성 계통선발을 위한 분자마커를 선발하였다. 이러한 과정을 통해 딸기 흰가루병 저항성 품종 육성을 위한 MAS(marker assisted selection) 체계 확립으로 내병성 육종효율 증진에 기여를 할 수 있을 것으로 기대된다.

키워드

집단분리분석법

Podosphaera aphanis

분자표지이용 선발법

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Information

Publisher :KOREAN SOCIETY FOR HORTICULTURAL SCIENCE
Publisher(Ko) :한국원예학회
Journal Title :Horticultural Science and Technology
Journal Title(Ko) :원예과학기술지
Volume : 33
No :5
Pages :722-729
Received Date : 2015-08-06
Revised Date : 2015-06-01
Accepted Date : 2015-06-02
DOI :https://doi.org/10.7235/hort.2015.14133

Horticultural Science and Technology 원예과학기술지 ISSN:1226-8763(Print) 2465-8588(Online)

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