Journal of The Korean Society of Grassland and Forage Science (한국초지조사료학회지)

The Korean Society of Grassland and Forage Science (한국초지조사료학회)
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Effect of Callus Type and Antioxidants on Plant Regeneration and Transformation of Tall Fescue

캘러스의 형태와 항산화물질 첨가가 톨 페스큐의 식물체 재분화와 형질전환효율에 미치는 영향

  • Lee Ki-Won (Division of Applied Life Science, College of Agriculture & Life Science, Gyeongsang National University) ;
  • Lee Sang-Hoon (Division of Applied Life Science, College of Agriculture & Life Science, Gyeongsang National University) ;
  • Kim Do-Hyun (Division of Applied Life Science, College of Agriculture & Life Science, Gyeongsang National University) ;
  • Lee Dong-Gi (Division of Applied Life Science, College of Agriculture & Life Science, Gyeongsang National University) ;
  • Won Sung-Hye (Division of Applied Life Science, College of Agriculture & Life Science, Gyeongsang National University) ;
  • Lee Hyo-Shin ;
  • Lee Byung-Hyun (Division of Applied Life Science, College of Agriculture & Life Science, Gyeongsang National University)
  • 이기원 (경상대학교 농업생명과학대학 응용생명과학부 낙농학) ;
  • 이상훈 (경상대학교 농업생명과학대학 응용생명과학부 낙농학) ;
  • 김도현 (경상대학교 농업생명과학대학 응용생명과학부 낙농학) ;
  • 이동기 (경상대학교 농업생명과학대학 응용생명과학부 낙농학) ;
  • 원성혜 (경상대학교 농업생명과학대학 응용생명과학부 낙농학) ;
  • 이효신 (국립산림과학원 생물공학과) ;
  • 이병현 (경상대학교 농업생명과학대학 응용생명과학부 낙농학)
  • Published : 2006.06.01
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Abstract

An efficient transformation system for the production of transgenic plants has been developed for tall fescue (Festuca arundinacea Schreb.) via Agrobacterium-mediated transformation of seed-derived callus. From the point of morphogenetic capacity, three types of callus were selected. High frequency of plant regeneration was obtained by selection of type II callus, and the plant regeneration frequency was 52.6% when embryogenic callus were cultured on the regeneration medium. Supplementation of the media with 10 mg/L $AgNO_3$ and 40 mg/L cysteine enhanced frequencies of plant regeneration up to 65.3%. The highest transformation efficiency was also obtained when type II callus were inoculated with Agrobacterium. Southern blot analysis of PCR products of transgenic plants demonstrated that transgenes were successfully integrated into the genome of tall fescue. Efficient regeneration system and transformation established in this study will be useful for molecular breeding of tall fescue through genetic transformation.

유용유전자 도입을 통한 신품종 톨 페스큐를 개발할 목적으로 Agrobacterium을 이용한 효율적인 식물체 재분화 및 형질전환에 미치는 몇 가지 요인을 조사하였다. 성숙종자로부터 유도된 캘러스를 형태에 따라 3가지 type으로 분류하였고 type II 캘러스는 유백색으로 녹색을 띠며 조직적으로 치밀한 상태이며 식물체로의 재분화효율이 52.6%로 가장 높게 나타났다. 또한 재분화 배지에 $AgNO_3$와 cysteine을 동시에 첨가해 준 경우 무첨가구에 비해 캘러스 유도율은 6.7%, 식물체 재분화율은 12% 씩 각각 증가하였다. 캘러스 type 별 형질전환 효율을 조사한 결과 type II 캘러스는 58.0%로 가장 높은 형질전환효율을 나타내었다. 형질전환체를 PCR 및 PCR-Southern blot 분석을 실시하여 본 결과 발현벡터의 T-DNA 영역이 형질전환 식물체의 genome에 성공적으로 도입되었음을 확인할 수 있었다. 본 연구를 통하여 확립된 효율적인 형질전환 시스템은 분자육종을 통한 신품종 톨페스큐의 개발에 유용하게 이용될 수 있을 것이다.

Keywords

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