Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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The Use of Glufosinate as a Selective Marker for the Transformation of Cucumber (Cucumis sativus L.)

오이의 형질전환을 위반 선발마커로서 Glufosinate의 이용

  • Cho Mi-Ae (Eugentech Inc., Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Song Yun-Mi (Eugentech Inc., Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Park Yun-Ok (Eugentech Inc., Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Ko Suck-Min (Eugentech Inc., Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Min Sung-Ran (Plant Cell Biotechnology Laboratory, Korea Research Institute of Bioscience and Biotechnology (KIRBB)) ;
  • Liu Jang-Ryol (Plant Cell Biotechnology Laboratory, Korea Research Institute of Bioscience and Biotechnology (KIRBB)) ;
  • Choi Pil-Son (Department of Medicinal Plant Resources, Nambu University)
  • Published : 2005.09.01
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Abstract

Agrobacterium tumefaciens-mediated cotyledonary explants transformation was used to produce transgenic cucumber. Cotyledonary explants of cucumber (c.v., Eunchim) were co-cultivated with strains Agrobaderium (LBA4404, GV3101, EHA101) containing the binary vector (pPTN289) carrying with CaMV 355 promoter-gus gene as reporter and NOS promoter-bar gene conferring resistance to glufosinate (herbicide Basta) as selectable marker. There was a significant difference in the transformation frequency depending Agrobacterium strains. The EHA101 of bacterial strains employed gave the maximum frequency (0.35%) for cucumber transformation. Histochemical gus and leaf painting assay showed that 15 individual lines were transgenic with the gus and bar gene. Southern blot analysis also revealed that the gus gene was successfully integrated into each genome of transgenic cucumber.

Agrobacterium과 자엽절편의 공동배양으로 박과작물인 오이의 형질전환체를 생산하였다. 오이 배양재료는 "은침"의 자엽절편을 사용하였으며, reporter유전자로서 gus유전자와 선발표지로서 bar 또는 nptII유전자로 각각 제작된 pPTN289와 pPTN290벡터를 GV3101, LBA4404, EHA101에 형질전환하여 공동배양하였다. 형질전환빈도는 Agrobacterium의 종류에 따라 현저한 차이가 있었으며, 특히 사용한 균주중 EHA101에서 0.35%로 가장 높았다. 선발배지에서 형성된 오이 식물체중 제초제 저항성 (12개체)과 paromomycin 저항성 (3개체)을 얻었고, 이들 모두 gus양성반응 나타냈다. Southern분석에 의하여 오이 형질전환체의 genome에 gus유전자가 도입되어 있음을 확인 하였다.

Keywords

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