Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Shoot Proliferation and Plant Regeneration from Suspension-Cultured Cells of Dianthus gratianopol

패랭이꽃속 Dianthus gratianopol의 현탁배양세포로부터 Shoot 증식과 식물체 재분화

  • Kim Joon-Chul (Division of Life Science, Kangwon National University)
  • Published : 2005.12.01
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Abstract

Conditions for efficient organogenesis and plant regeneration from Dianthus gratianopol suspension cultured cells were established. Shoot-forming calli of glossy surface, pale green and knobby type were selected from leaf explant-derived calli and were suspension-subcultured every week in CP liquid medium with 1.0 mg/L 2,4-D and 0.5 mg/L BAP. Combinations of 1.0 mg/L 2,4-D and 0.5 mg/L BAP, and 1.5 mg/L 2,4-D and 0.5 mg/L BAP were effective for the induction of regenerative callus from the suspension cultured cell clusters. Multiple shoot primordia were initiated from the green spots of these regenerative callus and formed shoots on MS medium with 1.0 mg/L TDZ and 0.5 mg/L PAA. Shoot regeneration frequency (calli regenerating at least one shoot) was about 87%. For plant regeneration, proliferated shoots were excised and transferred to MS medium with 0.1 mg/L NAA for root initiation after 9 weeks of culture. The regenerants were potted in soil and formed the flowering buds and petals. Also, adventitious shoots were formed from the excised green shoot primordia of regenerative callus and these shoots proliferated successfully and regenerated to whole plants.

패랭이꽃속 Dianthus gratianopol 잎절편체로부터 캘러스를 유기하여 표면에 광택이 있고 치밀하게 배열되고 연한 녹색의 shoot-forming 캘러스를 선별하였다. 유기된 캘러스를 잘게 자르고 1.0 mg/L 2,4-D와 0.5 mg/L BAP가 포함된 CP 액체배지에서 7일 간격으로 계대배양하여 균일한 SF형태의 조직이 치밀한 세포괴를 얻을 수 있었고 이들 세포괴를 1.0 mg/L 2,4-D와 0.5 mg/L BAP, 1.5 mg/L 2,4-D와 0.5 mg/L BAP의 MS고체배지에서 배양했을 때 재분화능력이 있는 캘러스를 유도할 수 있었다. 이들 캘러스는 1.0mg/L TDZ과 0.5 mg/L PAA가 포함된 MS재분화배지에 치상하였더니 녹화되기 시작하여 배양 4주후부터는 다량의shoot 원기가 형성되고 shoot증식이 가능하였으며 평균 87%의 shoot 형성률을 보였다. 배양 9주후 절단된 shoot는 0.1 mg/L NAA가 첨가된 배지에서 뿌리를 형성하여 화뢰와 꽃을 갖고 있는 완전한 식물체로 재분화되었다. 또한 재분화 능력이 있는 캘러스 정단 원기를 절단한 절편체는 1.0 mg/L TDZ 과 0.5 mg/L PAA가 포함된 MS재분화 배지에서 직접 부정아를 형성하였고 이들 부정아로부터 다량의 shoot 증식이 가능하여 식물체를 효과적으로 얻을 수 있었다.

Keywords

References

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