Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Efficient shoot regeneration using cotyledon explants and Agrobacterium tumefaciens - mediated transformation of bottle gourd (Lagenaria sicraria Standl.)

박 자엽조직을 이용한 효율적인 식물체 분화와 Agrobacterium에 의한 형질전환

  • Kim, Soo-Yun (Vegetable Research Division, National Institute of Horticultural & Herbal Science, Department of Life Sciences, RDA) ;
  • Ahn, Yul-Kyun (Vegetable Research Division, National Institute of Horticultural & Herbal Science, Department of Life Sciences, RDA) ;
  • Huh, Yun-Chan (Vegetable Research Division, National Institute of Horticultural & Herbal Science, Department of Life Sciences, RDA) ;
  • Lee, Hye-Eun (Vegetable Research Division, National Institute of Horticultural & Herbal Science, Department of Life Sciences, RDA) ;
  • Kim, Do-Sun (Vegetable Research Division, National Institute of Horticultural & Herbal Science, Department of Life Sciences, RDA)
  • 김수윤 (농촌진흥청 국립원예특작과학원 채소과) ;
  • 안율균 (농촌진흥청 국립원예특작과학원 채소과) ;
  • 허윤찬 (농촌진흥청 국립원예특작과학원 채소과) ;
  • 이혜은 (농촌진흥청 국립원예특작과학원 채소과) ;
  • 김도선 (농촌진흥청 국립원예특작과학원 채소과)
  • Received : 2014.08.06
  • Accepted : 2014.09.22
  • Published : 2014.09.30
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Abstract

This study were carried out for selection of proper transformation variety and development of efficient regeneration and transformation methods. The number of shoot in commercial varieties of gourd plant were 0 ~ 7.3. and fusarium wilt resistant pure lines were 2.0 ~ 6.5 per dish containing on MS medium supplemented with 3 mg/L BA. The shoot regeneration frequency of fusarium wilt resistant pure lines were wide variation on the deviation. The expression of GFP was high 67% and 100% at the co-cultivation with Agrobacterium. The effective shoot regeneration plant hormone were combination BA and 2,4-D. The number and elongation condition of shoot was good after 4 weeks change with MS medium supplemented with 1 mg/L BA. Effective callus production plant hormone were combination of 3 mg/L BA and 0.1 mg/L 2.4-D.

본 연구는 대목용으로 이용되는 박에 있어서 형질전환에 이용 가능한 품종을 선발하고 재분화 효율 및 형질전환 효율을 높이기 위하여 수행되었다. 3 mg/L BA가 첨가된 MS배지에서 덩굴마름병 저항성 순계계통의 평균 신초형성 수는 0 ~ 7.3개였으며 3종류의 상용 품종의 신초형성 수는 2.0 ~ 6.5개였다. 덩굴마름병 저항성 순계 계통의 재분화율은 품종 간 편차가 상용품종에 비하여 컸다. 형질전환 효율을 검정하기 위해 GFP와 gus발현을 조사한 결과 공동배양 과정 중에 GFP가 67%와 100% 로 높은 일시적인 발현을 보였다. 식물체 재분화는 BA와 2.4-D를 조합한 배지조합에서 자엽의 절단부위에서 신초가 발생하였으며, 4주 후 BA를 단독으로 1 mg/L 처리한 배지에서 한 개의 절편체에서 발생하는 신초의 개수 및 신초의 신장도 좋았다. 캘러스의 발생은 1 mg/L BA와 2,4-D를 0.1 mg 첨가한 배지에서 가장 활발하게 분화하였다.

Keywords

References

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