Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Development of transgenic strawberry plants expressing monellin, a sweet protein

감미단백질 모넬린 발현 딸기 형질전환 식물체 개발

  • Min, Sung Ran (Plant Systems Engineering Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Ko, Suk Min (Subtropical Horticulture Research Institute, Jeju National University) ;
  • Lyu, Jae Il (Plant Systems Engineering Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Park, Ji Hyun (Plant Systems Engineering Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Yi, So Young (Plant Systems Engineering Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Lee, In-Ha (Nonsan Strawberry Experiment Station, Chungcheongnam-do Agricultural Research & Extension Services) ;
  • Kim, Hyun Sook (Nonsan Strawberry Experiment Station, Chungcheongnam-do Agricultural Research & Extension Services) ;
  • Kim, Tae Il (Nonsan Strawberry Experiment Station, Chungcheongnam-do Agricultural Research & Extension Services) ;
  • Choi, Pil Son (Department of Oriental Pharmaceutical Development, Nambu University) ;
  • Jeong, Won-Joong (Plant Systems Engineering Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Kim, Suk Weon (Microbiological Resources Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Kim, Jonghyun (Division of Life Science, Korea Basic Science Institute) ;
  • Liu, Jang R. (Dept. of New Biology, Daegu Gyeongbuk Institute of Science & Technology)
  • 민성란 (한국생명공학연구원 식물시스템공학연구센터) ;
  • 고석민 (제주대 아열대원예산업연구소) ;
  • 유재일 (한국생명공학연구원 식물시스템공학연구센터) ;
  • 박지현 (한국생명공학연구원 식물시스템공학연구센터) ;
  • 이소영 (한국생명공학연구원 식물시스템공학연구센터) ;
  • 이인하 (충청남도농업기술원 논산딸기시험장) ;
  • 김현숙 (충청남도농업기술원 논산딸기시험장) ;
  • 김태일 (충청남도농업기술원 논산딸기시험장) ;
  • 최필선 (남부대 한방제약개발학과) ;
  • 정원중 (한국생명공학연구원 식물시스템공학연구센터) ;
  • 김석원 (한국생명공학연구원 미생물자원센터) ;
  • 김종현 (한국기초과학지원연구원 생명과학연구부) ;
  • 유장렬 (대구경북과학기술원 뉴바이올로지 전공)
  • Received : 2015.04.23
  • Accepted : 2015.07.02
  • Published : 2015.09.30
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Abstract

Leaf discs from 'Yeobong' and 'Maehyang' strawberry plants were used as explants for transformation. The Agrobacterium tumefaciens strain EHA105 harboring the monellin gene under the control of the CaMV 35S promoter was used in co-cultivation experiments. The frequencies of callus formation and plant regeneration from leaf explants after co-cultivation in 'Yeobong' were higher than those of 'Maehyang'. These transgenic plants showed normal growth patterns and flowering. PCR and Southern hybridization confirmed that 1 to 2 copies of the monellin gene were integrated into genome of the transgenic strawberry plants. Northern blot analysis confirm that the transcripts were expressed in transgenic strawberry plants. Although long-term subcultured transgenic strawberry plants showed a phenomenon to escape the transgene, the transformation system established in this study provides new opportunities for genetic improvement of strawberry plants.

'여봉'과 '매향' 딸기 식물체로부터 잎 절편을 형질전환 재료로 이용하였다. CaMV 35S promoter에 모넬린 유전자가 연결된 pGA482-pS1D 벡터가 들어있는 아그로박테리움 EHA105 균주를 매개로 형질전환을 수행하였다. 공동 배양 후 잎 절편체로부터 캘러스 형성과 식물체 재분화율은 '여봉' 품종이 '매향' 품종보다 높았으며 이들 형질전환 식물체는 정상적으로 생육하여 개화하였다. PCR 및 Southern blot 분석을 통해 1-2 카피의 모넬린 유전자가 형질전환 딸기 식물체에 도입되었음을 확인하였으며, Northern blot 분석을 통하여 두 품종에서 모두 모넬린 유전자가 발현됨을 확인하였다. 비록 장기간 계대배양된 이들 딸기 형질전환 식물체에서는 모넬린 유전자가 escape 되는 경향을 보였지만, 본 연구에서 확립된 형질전환 시스템은 딸기의 유전적 개량을 위한 새로운 기회를 제공할 수 있을 것이다.

Keywords

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