Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Plant Regeneration and Mutagenesis from Organogenic Callus of Dianthus Distributed in Gangwon Province

강원지역 패랭이꽃속의 캘러스로부터 식물체 재분화와 돌연변이체 유발

  • Published : 2003.03.01
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Abstract

Useful Dianthus species were collected and selected from two native and seven foreign species distributed in Gangwon province. For in vitro breeding,. callus was induced from the explants of apical meristem, leaf, stem and the in vitro adventitious shoots on MS basal medium with 2.0 mg/L 2,4-D and 0.5 mg/L BA at 27$^{\circ}C$ under continuous light. After 3 weeks of culture, calli initiated the most highly from the leaf explants of D. chinensis Organogenic calli were able to be selected from the adventitious shoot-derived calli. For shoot regeneration, these organogenic calli were cultured on MS medium with the combination of 0.1 mg/L NAA＋1.0 mg/L BA under continuous light. Multiple shoots were proliferated with low frequency (about 30%) from those adventitious shootderived calli. Also, shoots initiated directly from the adventitious shoot explants without callus formation at high frequency of 52% when cultured on N6 medium containing 0.1 mg/L NAA and 1.0 mg/L BA in D. gratianopol. Multiple shoots and plantlets grew well and rooted on MS medium supplemented with 0.1 mg/L NAA. Regenerants with well-developed roots were transferred to 8-cm pots containing vermiculite at 85% relative humidity and 27$^{\circ}C$ These plantlets were acclimatized in artificial soil mixture and transferred to the greenhouse for flowering with normal phenotypes. M28 Mutant line was selected with white flowers from 0.03M EMS-treated organogenic calli derived from in vitro adventitious shoot explants of D. chinensis and set seeds.

패랭이꽃속의 분포와 개발을 위한 기초자료로 강원도에서 자생 2종과 수집가능한 7 외래종에 대한 기내배양체의 특성을 조사하였다. 캘러스의 유도를 위해 조직부위별 다양한 절편체들을 2,4-D. NAA, BA가 농도별로 처리된 MS 배지에 치상한 후, 27$^{\circ}C$ 광조건과 2.0mg/L 2,4-D와 0.5 mg/L BA의 조합배지에서 배양했을 때 3주 후 절단면에서 callus가 형성되기 시작하였으며, 패랭이꽃 (Dianthus chinensis)의 잎절편체에서 가장 양호하였다. Organogenic 캘러스의 선발은 기내에서 증식된 adventitious shoot의 절편체 유래 캘러스로부터 가능했으며 이들 캘러스를 1.0 mg/L BA와 0.1 mg/L NAA가 조합된 배지에 치상하였을 때 약 30% 정도의 shoot 형성률을 보였고 multiple shoot의 분화를 관찰할 수 있었다. In vitro에서 증식된 adventitious shoot 절편체를 1.0 mg/L BA와 0.1 mg/L NAA가 포함된 N6 배지에 치상했을 때, 캘러스 단계를 거치지 않고 직접 shoot이 형성되었고 D. gratianopol 유래 adventitious shoot 절편체는 52%의 높은 shoot 형성률을 보였다. 이들 multiple shoot로부터 재분화된 소식물체는 0.1 mg/L NAA가 포함된 MS 배지에서 뿌리가 유도되고 vermiculite가 담긴 pot에 이식 후 85% 습도하에서 활착되어 정상적인 개화된 개체를 획득할 수 있었다. 0.03 M EMS 처리된 패랭이꽃의 adventitious, shoot 절편체 유래 캘러스로부터 힌꽃의 패랭이꽃의 도연변체식물 (M23)이 분화되었으며 종자 형성이 가능하였다.

Keywords

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