Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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The use of cotyledonary-node explants in Agrobacterium tumefaciensmediated transformation of cucumber (Cucumis sativus L.)

Agrobacterium에 의한 오이 형질전환에서 자엽절 절편의 이용

  • Jang, Hyun-A (Department of Oriental Pharmaceutical Development, Nambu University) ;
  • Kim, Hyun-A (Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Kwon, Suk-Yoon (Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Choi, Dong-Woog (Department of Biological Education, Jeonnam National University) ;
  • Choi, Pil-Son (Department of Oriental Pharmaceutical Development, Nambu University)
  • 장현아 (남부대학교 약용식물형질전환연구소) ;
  • 김현아 (한국생명공학연구원) ;
  • 권석윤 (한국생명공학연구원) ;
  • 최동욱 (전남대학교 생물교육과) ;
  • 최필선 (남부대학교 약용식물형질전환연구소)
  • Received : 2011.06.29
  • Accepted : 2011.07.30
  • Published : 2011.09.30
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Abstract

Agrobacterium tumefaciens-mediated cotyledonary-node explants transformation was used to produce transgenic cucumber. Cotyledonary-node explants of cucumber (Cucumis sativus L. cv., Eunsung) were co-cultivated with Agrobacterium strains (EHA101) containing the binary vector (pPZP211) carrying with CaMV 35S promoter-nptII gene as selectable marker gene and 35S promoter-DQ gene (unpublished data) as target gene. The average of transformation efficiency (4.01%) was obtained from three times experiments and the maximum efficiency was shown at 5.97%. A total of 9 putative transgenic plants resistant to paromomycin were produced from the cultures of cotyledonary-node explants on selection medium. Among them, 6 transgenic plants showed that the nptII gene integrated into each genome of cucumber by Southern blot analysis.

Agrobacterium과 자엽절 절편 공동배양으로 오이 형질전환체를 생산하였다. 오이 자엽절 절편 (c.v. Eunsung)은 선발 마커로서 nptII유전자와 유용유전자로서 DQ유전자가 포함된 pPZP211를 EHA101에 형질전환하여 공동 배양하였다. 3회 반복실험으로부터 평균 형질전환 빈도는 4.01%를, 최대 빈도는 5.97%를 보여 주었다. Paromomycin항생제 저항성을 갖는 9개의 식물체를 선발과정을 통해 얻었으며, Southern blot 분석에 의해 6개 식물체의 genome에 nptII유전자자 안정적으로 도입되어 있음을 확인할 수 있었다.

Keywords

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