Korean Journal of Plant Tissue Culture (식물조직배양학회지)

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Effects of Dimethylsulfoxide on the Cell Wall Regeneration and Cell Division of Protoplasts Isolated from Panax ginseng Callus

인삼 캘러스 원형질체의 세포분열과 세포벽 재생에 미치는 Dimethylsulfoxide의 효과

  • 이석찬 (성균관대학교 생명공학부 유전공학과) ;
  • 이규배 (조선대학교 사범대학 생물교육학과) ;
  • 박종범 (신라대학교 자연과학과군 생명과학과)
  • Published : 2000.11.01
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Abstract

Isolated protoplasts from ginseng (Panax ginseng C. A. Meyer) callus tissue were cultured in modified MS media supplemented with various concentrations of dimethylsulfoxide (DMSO). The cell wall regeneration rate and cell division efficiency of the protoplasts were increased significantly by 1% DMSO treatment. However, there was no difference in the viability of protoplasts between the DMSO treatment and non-treatment. Transmission electron microscopy revealed that the microtubules were oriented in parallel manner to the plasmalemma after 3 days of culture in medium with 1% DMSO. Further, interconnected cellulose microfibrils were observed on the outer surface of the 3-day-cultured protoplasts by scanning electron microscopy These structures shown by electron microscopy were not observed in protoplasts cultured on DMSO-free media. This studies indicates that DMSO supplemented in culture media seemed to stimulate the cell wall regeneration and cell divisions of protoplasts by forming microtubule organizing centers (MTOC).

인삼 (Panax ginseng C. A. Meyer) 캘러스 조직으로부터 분리한 원형질체를 DMSO가 O％에서 8％까지 여러 가지 농도로 첨가된 원형질체 배양배지 (1 mg/L 2,4-D, 4 mg/L NAA, 1 mg/L BAP, 0.4 M mannitol 및 0.8% agar가 첨가된 MS배지)에서 배양하였다. DMSO가 첨가되지 않은 배지에서 배양된 원형질체의 세포벽 재생률은 약 62%이고 세포분열빈도는 약 7％이었다. 반면에, 1% DMSO가 첨가된 배지에서 배양된 원형질체의 세포벽 재생률과 세포분열빈도는 각각 약 83%와 약 27%로 높게 나타났다. 그러나 배양된 원형질체의 세포활성은 배지에 첨가된 DMSO의 유무나 농도와는 관계없이 모든 배지에서 83∼88%로 차이를 보이지 않았다. 1% DMSO가 첨가된 배지에서 3일 동안 배양된 원형질체를 투과전자현미경으로 관찰하면 원형질막 근처에 평행으로 배열하고 있는 미소관들이 관찰되었다. 또한 원형질막 표면에는 세포벽 성분인 cellulose fibril들이 연결되어 다발을 형성하고 있는 것이 주사전자현미경으로 관찰되었다. DMSO가 첨가되지 않은 배지에서 배양된 원형질체에서는 이러한 전자현미경적 구조들이 관찰되지 않았다. 원형질체 배양배지에 첨가된 DMSO는 미소관편제센타 (MTOC)의 형성에 의하여 세포벽 재생과 세포분열을 유도하는 것으로 생각된다.

Keywords

References

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