카네이션 (Dianthus caryophillus)의 색소 발현체계 분석
Characterization of flavonoids specific gene expression in the petals of Dianthus caryophyllus (carnation)
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초록
본 연구에서는 색소유전자의 전이를 통하여 새로운 색소 발현체계를 가진 품종을 육종하기 위한 기초연구로 카네 이션 (Dianthus caryophyllus)의 꽃봉오리로부터 cDNA-library 를 합성하였고 페튜니아의 CHS, FHT, DFR, ANS 유전자 를 probe로 사용하여 anthocyanin 합성경로의 중요 유전자 들인 카네이션의 CHS, FHT, DFR, 그리고ANS 유전자를 분리하였다. 수집한 카네이션 품종 및 계통으로부터 돌 연변이체를 선발하기 위해 분리한 유전자들를 probe로 사용하여 Northern blot 분석을 수행하여 DFR 돌연변이체 로는 금별, 은별, Ballatyne, Crystal, Eugenia, Koreno, Imp. white sim., W-crystal, W-alpine 및 White charotte등 10개 계 통이 선발되었고 이 중 Eugenia, Imp. white sim. 및 W-alpine 은 DFR과 ANS의 이중돌연변이체로 밝혀졌고 Koreno는 DFR과 CHS의 이중돌연변이로 사려되며, 금별, Ballatyne 은 DFR, CHI의 이중 돌연변이체로 판명되어 목적식물에 서 제외되고 최종적으로 은별, Crystal, West crystal, White charotte 그리고 이미 유전적으로 DFR 돌연변이체로 선발 된 Verginie를 포함하여 총 5계통의 카네이션이 목적식물 로 선발되었다. 선발된 DFR 돌연변이체의 확인을 위하 여 유전자의 효소활성을 측정해 본 결과 5품종의 카네이 션 모두 DFR 활성을 가지고 있지 않음을 확인하였다
Abstract
This study aimed to develop carnation cultivars with new coloring system. We used four genes of Petunia hybrida - chalcone synthase (CHS), flavanone 3-hydroxylase (FHT), dihydroflavonol 4-reductase (DFR), and anthocyanidin synthase (ANS) - as probes, in order to isolate four genes from carnations (Dianthus Caryophyllus). The isolated genes were used as probes in order to select mutants out of collected carnations, using Northern blot analysis. The Northern blot analysis revealed 10 DFR mutants - Gumbyul, Eunbyul, Ballatyne, Crystal, Eugenia, Koreno, Imp. White Sim, West Crystal, White Alpine, and White Charotte. Six among the selected 10 cultivarswere excluded from the target cultivars, because Eugenia, Imp. White Sim, and White Alpine were proved to be double mutants of DFR and ANS, Koreno was considered to be a double mutant of DFR and CHS, and Gumbyul and Ballatyne were proved to be double mutants of DFR and CHI (Chalcone isomerase). Consequently, we selected five DFR mutants, including Virginie, which was already selected as a DFR mutant. Finally, we measured DFR activities in order to confirm the selection, and the results showed that all of the five cultivars - Eunbyul, Crystal, West Crystal, White Charotte, and Virginie - had got no DFR activity.
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