Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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A simple mid-term preservation method (SMPM) of plant callus under low temperature conditions

저온 보존을 이용한 간편 중기 식물캘러스 저장법

  • Park, Sung-Chul (Biological Resource Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Park, Su Hyun (Biological Resource Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Kim, Soyoung (Biological Resource Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Jeong, Yu Jeong (Biological Resource Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Kim, Cha Young (Biological Resource Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Jeong, Jae Cheol (Biological Resource Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB))
  • 박성철 (한국생명공학연구원 생물자원센터) ;
  • 박수현 (한국생명공학연구원 생물자원센터) ;
  • 김소영 (한국생명공학연구원 생물자원센터) ;
  • 정유정 (한국생명공학연구원 생물자원센터) ;
  • 김차영 (한국생명공학연구원 생물자원센터) ;
  • 정재철 (한국생명공학연구원 생물자원센터)
  • Received : 2022.09.23
  • Accepted : 2022.09.27
  • Published : 2022.09.30
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Abstract

The repeated monthly or weekly subculture of plant callus is labor intensive and increases the risk of somaclonal variation from the parental callus line. The most effective method for preserving plant callus is cryopreservation, which involves storage in liquid nitrogen. However, this method cannot be applied to the callus of different plant species in the same manner, so it is difficult to develop a standardized cryopreservation method. In addition, the survival rate of the frozen callus after thawing and the regeneration rate after survival are uncertain. Therefore, it is necessary to develop a method to extend the subculture interval of plant callus in an active state. In this study, active plant calli of various species without freezing was incubated at 15℃ for 4 to 12 weeks without subculture. After 12 weeks, 8 lines of plant callus grew less than 2-fold when cultured at 25℃, but at least 2 times as much when cultured at 15℃. Moreover, total antioxidant activity did not differ significantly between plant callus recovered at 25℃ after culturing at 15℃ or at 25℃. These results show that the subculture interval can be extended at a temperature of 15℃ without need for modified medium composition or additional processes. In addition, positive results in all calli of several plant species are expected to reduce labor as well as somaclonal variation by increasing the subculture.

식물 캘러스의 월간 또는 주간의 반복적인 계대배양은 노동 집약적이며 모 세포주로부터 somaclonal variation 발생 위험을 증가시킨다. 식물 캘러스를 보존하기위한 가장 효과적인 방법은 액체질소에 저장하는 초저온동결보존 방법이다. 하지만 초저온동결보존 방법은 동일한 방법으로 여러 식물의 캘러스에 적용할 수 없어 보존 방법 개발에 많은 어려움이 따른다. 또한 해동 후 냉동되어진 캘러스의 생존과 생존 후 재생 속도가 불확실 하다는 단점이 있다. 그러므로 활성 상태의 식물 캘러스의 계대배양 간격을 증가시키는 방법의 개발이 필요하다. 본 연구에서는 냉동과정 없는 활성상태의 다양한 종의 식물 캘러스를 계대배양 없이 4주에서 12주 동안 15℃에서 배양하였다. 25℃에서 12주간 배양한 8종류의 식물 캘러스들은 모두 2배 이하의 성장을 보였으나 15℃에서 12주간 배양 조건에서는 8종류의 식물 캘러스들은 최소 2배 이상의 성장을 하였다. 또한 15℃에서 배양 후 25℃에서 회복시킨 캘러스들 사이의 항산화 활성 역시 크게 변화하지 변하지 않았다. 이러한 결과는 배지조성이나 특별한 새로운 과정없이 15℃ 저온으로 계대배양 간격을 증가시킬 수 있음을 보여준다. 또한 여러 식물 종의 캘러스들 모두에서 긍정적인 결과는 여러 캘러스들에 동일한 방법으로 계대배양 간격을 증가시킴으로 노동력 감소는 물론 somaclonal variation을 상대적으로 줄여 줄 것으로 예상한다.

Keywords

Acknowledgement

본 연구는 KRIBB 기관고유사업(KGM5382212, KGM5282223) 및 과학기술정보통신부의 재원으로 한국연구재단 바이오·의료기술개발사업의(2020M3A9I4038354) 지원을 받아 수행된 연구임.

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