Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Development of herbicide-tolerant Korean rapeseed (Brassica napus L.) cultivars

한국 고유의 품종을 이용한 제초제 저항성 유채 개발

  • Kim, Hyo-Jin (Department of Bioenergy Science & Technology, Chonnam National University) ;
  • Lee, Hye-Jin (Department of Bioenergy Science & Technology, Chonnam National University) ;
  • Go, Young-Sam (Department of Plant Biotechnology, Chonnam National University) ;
  • Roh, Kyung-Hee (National Academy of Agricultural Science, Rural Development Administration) ;
  • Lee, Young-Hwa (Bioenergy Crop Research Center and National Institute of Crop Science, Rural Development Administration) ;
  • Jang, Young-Seok (Bioenergy Crop Research Center and National Institute of Crop Science, Rural Development Administration) ;
  • Suh, Mi-Chung (Department of Bioenergy Science & Technology, Chonnam National University)
  • 김효진 (전남대학교 바이오에너지공학과) ;
  • 이혜진 (전남대학교 바이오에너지공학과) ;
  • 고영삼 (전남대학교 식물생명공학부) ;
  • 노경희 (농촌진흥청 국립농업과학원) ;
  • 이영화 (농촌진흥청 국립식량과학원 바이오에너지작물센터) ;
  • 장영석 (농촌진흥청 국립식량과학원 바이오에너지작물센터) ;
  • 서미정 (전남대학교 바이오에너지공학과)
  • Received : 2010.08.31
  • Accepted : 2010.09.03
  • Published : 2010.09.30
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Abstract

An interest in the production of seed-oil based fuel and raw materials, which comes from renewable plant sources, has been intrigued by the phenomenon of global warming and shortage of fossil fuels. Rapeseed (Brassica napus) is the most important oilseed crop, which produces seeds with 40% oil. It is desirable to develop genetically modified rapeseed producing oils, which can be easily converted to biodiesel. As an initial step for development of genetically modified rapeseed for the production of biofuels or bio-based materials, Korean rapeseed cultivars, Naehan, Youngsan, Tammi and Halla, were analyzed. Four Korean rapeseed cultivars produce 32 to 40% oil of seed dry weight, which is rich in oleic acid (more than 60 mole%). The cotyledonary petioles of rapeseed cultivar, Halla, were transformed using Agrobacterium tumefaciens strain GV3101, carrying the uidA gene encoding $\beta$-glucuronidase (GUS) as a reporter gene and the phosphinothricin acetyltransferase (PAT) gene as a selectable marker. The stable integration of PAT gene in the genome of transgenic rapeseeds was confirmed by PCR analysis. Expression of uidA gene in various rapeseed organs was determined by fluorometric assay and histochemical staining. Transformation efficiency of a Korean rapeseed Halla cultivar was 10.4%. Genetic inheritance of transgenes was confirmed in $T_2$ generation.

화석에너지의 고갈과 지구 온난화 현상으로 인해 재생 가능한 식물자원으로부터 바이오에너지를 얻고자 하는 관심이 높아지고 있다. 이에 바이오디젤의 원료로 사용 되기 적합한 형질전환이 된 유채를 개발하기 위한 첫 단계로 한국 고유 유채 품종을 이용한 형질전환 체계를 구축하였다. 내한, 영산, 탐미, 한라 유채의 종자를 분양 받아 지방산 분석을 실시한 결과, 종자의 약 32-40% 식물성 오일이 포함되어 있었고, 그 중 올레인산의 함량은 60mole% 이상 존재하는 것으로 확인되었다. 그 중 오일 함량 및 올레인산 함량이 높고, 형질전환 효율이 비교적 높은 한라 유채품종이 그리고 $\beta$-glucuronidase (GUS)와 phosphinothricin acetyltransferase (PAT) 유전자가 포함된 pCAM-BIA3301 벡터가 도입된 Agrobacterium tumefaciens GV3101균주가 형질전환에 사용되었다. 형질전환이 된 유채 식물체는 제초제에 대한 내성, PCR을 이용한 PAT 유전자의 도입 여부 및 GUS 활성 분석을 통하여 선별하였다. 그 결과 한라 유채의 경우, 10. 4% 형질전환 효율을 보였고, 제초제 저항성이 다음 세대 ($T_1$ 식물체)로 안정되게 유전됨을 확인하였다. 이러한 연구는 바이오디젤 원료로 사용될 다양한 유채 품종에 교배를 통해 제초제 저항성 유전자를 쉽게 도입할 수 있는 가능성을 제시하였다.

Keywords

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