Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Cryopreservation of Embryogenic Callus in Sweetpotato cv. 'Yulmi'

고구마품종 '율미' 배발생 캘러스의 초저온 동결보존

  • Park, Jong-Suk (Research Institute of Bioindustry, College of Agriculture, Chonbuk National University) ;
  • Kim, Suk-Weon (Bioresources Center) ;
  • In, Dong-Su (Eugentech Inc., KRIBB) ;
  • Eun, Jong-Seon (Research Institute of Bioindustry, College of Agriculture, Chonbuk National University)
  • 박종숙 (전북대학교 농과대학 생물산업연구소) ;
  • 김석원 (한국생명공학연구원 생물자원센터) ;
  • 인동수 ((주)유진텍) ;
  • 은종선 (전북대학교 농과대학 생물산업연구소)
  • Published : 2003.03.01
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Abstract

Cryopreservation of embryogenic callus derived from apical meristem culture was attempted by slow prefreezing method (two-step method) with various cryoprotectants in sweetpotato cv. 'Yulmi' Precultured embryogenic calli on medium containing 10 mg/L ABA prior to slow prefreezing in liquid nitrogen indicated higher survival rate than 1.0 mg/L ABA preteatment. The cryoprotectant comprising 1.28 M DMSO in 0.4 M sucrose solution gave the best survival (over 46%) of sweetpotato cells exposed to liquid nitrogen as determined by TTC reduction and FDA staining method. Cryopreserved calli cultured on MS medium with 1.0 mg/L 2,4-D were grown for 4 weeks in the dark and induced embryos after another 4 weeks. They were subcultured on MS medium supplemented with 0.1 mg/L 2,4-D＋0.1 mg/L kinetin for 2 weeks and regenerated into normal plantlets in MS basal medium.

고구마 품종 '율미'를 이용해 2,4-D 1.0 mg/L가 첨가된 MS 배지에 정단분열조직배양을 통하여 발생된 배발생 캘러스를 실험재료로 여러 가지 동결보호제를 처리하여 2-step method 에 의해 동결보존을 실시하였다. ABA 10m/L가 포함된 배지에서 전처리된 배발생 캘러스는 ABA 1.0mg/L 처리구보다 액체질소에 저장 후 생존율이 더 높게 나타났다. TTC방법과 FDA 염색법을 통해 초저온 보존 후에 생존을 확인한 결과 10mg/L의 ABA를 전처리 한 0.4M sucrose가 포함된 1.28M DMSO 처리구에서 46.8%의 가장 높은 생존율을 나타냈다. 캘러스를 1.0mg/L 2,4-D가 포함된 MS 고체배지에서 암배양한 결과 배양 4주일 후부터 1.28 M DMSO 단독처리구에서 캘러스가 생장하는 것을 육안 관찰할 수 있었으며 배양 8주일 후에는 배가 발생하였고, 0.1 mg/L 2,4-D＋0.1 mg/L kinetin 혼용구에 2주일간 계대배양한 다음 MS기본배지에서 완전한 식물체로 재생되었다.

Keywords

References

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