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배추 원형질체 배양에 미치는 다양한 처리의 효과와 원형질체 유래 캘러스로부터 신초 재분화

Effects of a variety of treatments affecting Chinese cabbage protoplast culture, and plant regeneration from protoplast-derived callus

  • Han, Jeung-Sul (National Horticultural Research Institute, Rural Development Administration) ;
  • Yoon, Moo-Kyeong (National Horticultural Research Institute, Rural Development Administration) ;
  • Jeong, Mi-Hye (National Institute of Agricultural Science and Technology, Rural Development Administration)
  • 발행 : 2008.09.30

초록

본 연구에서는 배추 원형질체 배양에 있어 다양한 처리의 효과를 구명함으로써 배추 원형질체 배양을 통한 재분화 체계를 확립하고자 하였다. 배추 원형질체 분리 수율은 효소액, 사용된 조직 및 효소액 처리 시간에 따라 달리 나타났는데, 효소액 처리 약 7시간 이후부터는 분리되는 원형질체의 수가 크게 증가하지 않았다. 따라서 효소 자체에 의한 부정적 영향을 고려하였을 때 최장 7시간 동안 효소처리를 하는 것이 적정한 것으로 판단하였다. 떡잎유래 원형질체를 2,4-D 1.0 mg/L, NAA 0.5 mg/L 및 BA 1.0 mg/L 첨가 액체배지에서 배양하였을 때 초기 세포분열과 이어지는 micro-callus 생장이 가장 양호하였다. 다양한 배추 인자형에 대한 검토 결과 본 연구의 배양조건 하에서는 초기 세포분열과micro-callus 생장이 배추 인자형에 따라 뚜렷하게 영향을 받는 것으로 나타났다. 노랑봄 품종의 떡잎에서 분리한 원형질체로부터 생장한 micro-callus 현탁액을 zeatin 0.2 mg/L외 spermidine 0.1 mM이 첨가된 반고체 MS 배지에 plating하였을 때 가장 왕성하게 callus colony가 형성 및 생장하였다. 이들 callus colony는 BA 5.0 mg 및 NAA 1.0 mg/L가 첨가된 배지에서 4.3%의 빈도로 재분화 신초가 유도되었다 얻어진 신초는 PGR 무첨가 MS배지에서 손쉽게 발근하였고 발근된 식군체는 기외에서 정상 식물체로 생장하였다. 본 연구의 결과는 배추 단독 원형질체 배양을 통한 고빈도 식물체 재분화 연구에 중요한 정보를 제공함으로써 유용하게 활용될 것으로 사료된다.

Here we describe a procedure for Chinese cabbage protoplast culture and effects of various treatments. Chinese cabbage protoplasts were isolated from different parts of young seedlings as using an enzyme mixture, of which yield was maximized in seven hours around after digestion. The highest rate of initial cell division followed by micro-callus formation was obtained in the medium with 1.0 mg/L 2,4-D, 0.5 mg/L NAA, and 1.0 mg/L BA when the cotyledon-derived protoplasts were cultured. Initiation of cell division and micro-callus proliferation significantly depended upon Chinese cabbage genotype under a same culture circumstances. The micro-calli developed from cotyledon tissue of Norang-Bom cultivar successfully grew toward callus colonies on the solidified medium with 0.2 mg/L zeatin and 0.1 mM spermidine. The callus colonies generated de novo shoots at the maximum frequency of 4.3% on the medium with 5.0 mg/L BA and 1.0 mg/L NM. Our results might be helpful for further studies to enhance the regeneration efficiency in Chinese cabbage protoplast culture.

키워드

참고문헌

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피인용 문헌

  1. High frequency somatic embryogenesis through leaf explant-derived callus culture in Muscari armeniacum cv. 'Early Giant' vol.39, pp.1, 2012, https://doi.org/10.5010/JPB.2012.39.1.069