포도나무의 내한성 관련 SCAR 분자표지 개발
Development of SCAR Markers Linked to Cold Resistance for Marker-assisted Selection of Grapevines
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포도 신품종 육성에서 내한성 계통의 조기 선발을 위하여, 다양한 품종을 대상으로 randomlyamplified polymorphic DNA(RAPD)를 수행하고, 그 결과를 바탕으로‘Campbell Early’x ‘Kaiji’,‘Golden Muscat’x‘Tamnara’,‘Campbell Early’x‘Neo Muscat’와‘Alden’x‘Kaiji’의 포도교배조합 실생계통을 이용하여 내한성 관련 sequence characterized amplified region(SCAR) 분자표지를 개발하였다. 400여개의 primer를 사용하여 bulk집단에서의 RAPD 분석을 통해서 6개의 UBCrandom primer를 최종적으로 선발하였고, RAPD 산물을 클로닝한 후 염기서열을 분석하였다. 6개의SCAR primer (UBC123-428, UBC196-353, UBC221-503, UBC317-800, UBC342-500, UBC344-451)를 제작하고 교배조합 실생에 재검정한 결과 내한성과 관련한 특이밴드를 확인할 수 있었다. 포도나무의 내한성과 관련한 특이적인 밴드출현양상은 수피의 탄수화물함량, 전해질전도도, 포장에서의 신초생존율과 유사한 양상을 보였다. 본 연구에서 개발한 SCAR marker는 포도의 염색체상에서 위치를 확인할 수 있었으며, 그 주변에는 주로 cytochrome P450, glutathione S-transferase, leucine-rich repeat,pleiotropic drug resistance 12, ribosomal protein 등의 유전자가 분포하였다. 본 연구에서 개발된포도 내한성 관련 SCAR marker는 내한성 포도 품종의 조기선발과 육종효율 증진에 기여할 수 있을 것으로 기대된다.
Abstract
To improve the efficiency for breeding new grape cultivars tolerant to coldness, sequencecharacterized amplified region (SCAR) markers were developed from randomly amplifiedpolymorphic DNA (RAPD) markers for cold tolerance in seedlings derived from the crossesof ‘Campbell Early’ x ‘Kaiji’, ‘Golden Muscat’ x ‘Tamnara’, ‘Campbell Early’ x ‘NeoMuscat’, and ‘Alden’ x ‘Kaiji’. Bulked segregant analysis (BSA) using 400 primers wereconducted and 6 UBC random primers showing polymorphisms were finally selected. RAPDproducts with 6 primers were cloned and sequenced. The amplification with 6 SCAR primers(UBC123-428, UBC196-353, UBC221-503, UBC317-800, UBC342-500, and UBC344-451) developedfrom RAPD analysis were tested and resulted in the amplification of specific fragments ingrapevine cross seedlings. Detection of specific fragments related with cold hardiness showedsimilar patterns in cold tolerance tested by investigation of total carbohydrate content in vine bark skin, electrolytic conductance and bud breakage in grapevine cross seedlings. SCARmakers developed in this study were mapped in chromosomes which genes such ascytochrome P450, glutathione S-transferase, leucine-rich repeat, pleiotropic drug resistance 12,and ribosomal protein were located. The results suggested that new SCAR markers could beused for increasing the efficiency for the selection of cold-hardy seedlings in grape breedingprograms.
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