대반하[Pinellia tripartita(Blume) Schott]의 최적기내배양 및 재분화 조건 규명을 통한 대량번식
Mass Propagation of Pinellia tripartita(Blume) Schott through Optimization of In vitro Cultures and Regeneration System
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대반하[大半夏: Pinellia tripartita(Blume) Schott]는 천남성과에 속하는 초본성 식물로써, 중요 한약재인 반하[半夏: Pinellia ternata(Thunb.) Makino]의 국내 자생 동속 종으로 활용가치가 높은 식물이나 반하에 비해 생리학 및 유전학 등 관련 연구가 거의 진행되지 않았다. 본 연구는 대반하의 소괴경형성 및 분화에 적합한 최적 배지와 식물생장조절물질의 농도를 규명하고, 유전적인 안전성 및 토양환경 순화조건을 검증하여 향후 반하의 대체 약재로 활용시 이용가능한 배양묘 대량생산조건을 확립하기위하여 수행되었다. 자생지에서 채집한 대반하는 소독 후 MS 기본배지에서 배양하여 기내식물체를 확보하였고, 소괴경 분화와 비대 최적배지 규명을 위해서 10종류의 다양한 식물배지에서 6주간 배양 한결과, Nitsch & Nitsch(NN) 배지에서 가장 효과적이었다. 기내 배양 시 발생한 캘러스를 이용하여 식물체 최적 재분화 조건을 규명하기 위하여, 명/암 반응과 BA 호르몬의 다양한 농도별(0.5ppm-3.0ppm)처리를 통해서 4주 동안 배양한 결과, 명상태에서 BA 1.0 ppm이 첨가된 배지에서 가장 효과가 높았다. 최적 기내배양 및 재분화 개체는 2주 후 토양으로 이식하여 환경적응을 유도한 후 배양묘 대량 생산조건을 설정하였다. 기내배양 및 재분화를 통해 생산된 배양묘의 유전적 안전성을 검증하기 위해서각 증식 단계별 식물조직 및 캘러스 시료의 RAPD genomic profile을 분석한 결과, 원 식물체와 각 단계별 개체의 유전적 안전성이 증명되었다.
Abstract
Pinellia tripartita(Blume) Schott is a medicinal herbaceous plant belonging to the Araceae,Korea which is a candidate plant species for contrary or alternative medicine of Pinelliaternata(Thunb.) Makino. The medicinal parts of P. tripartita(Blume) Schott are used as roottuber. This study investigated the optimal media and the concentrations of plant growth regulatorfor small tuber formation and differentiation of P. tripartita(Blume) Schott. It can be used as acandidate for substitute or alternative medicinal plants as Pinelliae Tube and contributed geneticstability with the guaranteed mass production. After picking samples from natural habitat, theywere cultured in MS basal medium. To optimize in vitro culture and to improve biomass oftubers, 10 variety of plant culture media were used for 6 weeks. The most effective mediumwas Nitsch & Nitsch(NN) medium. In order to identify the optimum plant regeneration condition caused by a callus culture, most efficient response from various concentrations(0.5ppm- 3.0ppm) of BA was in 1.0 ppm of BA after 4 weeks. After 2 weeks, the regenerated plantswere transplanted into soils for environmental adaptation. To verify the genetic stability in eachgrowth stage of plant tissues and callus, the samples were extracted DNA and performed byRAPD analysis. All stages of plant tissues and callus were genetically stable.
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