Korean Journal of Plant Resources (한국자원식물학회지)

The Plant Resources Society of Korea (한국자원식물학회)
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Transformation of 'Ilmibyeo' using pCAMBIA 1300 and Microstructural Investigation of Leaves

pCAMBIA 1300 벡터를 이용한 일미벼의 형질전환 및 잎의 전자현미경적 관찰

  • Guo, Jia (School of Biotechnology, Kangwon National University) ;
  • Seong, Eun-Soo (School of Biotechnology, Kangwon National University) ;
  • Kim, Young-Hwa (Department of Plant Biotechnology, Dongguk University) ;
  • Jo, Hye-Jeong (School of Biotechnology, Kangwon National University) ;
  • Cho, Joon-Hyeong (Department of Plant Biotechnology, Dongguk University) ;
  • Wang, Myeong-Hyeon (School of Biotechnology, Kangwon National University)
  • 곽가 (강원대학교 생명공학부) ;
  • 성은수 (강원대학교 생명공학부) ;
  • 김영화 (동국대학교 식물생명공학과) ;
  • 조혜정 (강원대학교 생명공학부) ;
  • 조준형 (동국대학교 식물생명공학과) ;
  • 왕명현 (강원대학교 생명공학부)
  • Published : 2007.10.30
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Abstract

The argE gene of E.coli was introduced into #Ilmibyeo# cultivar of rice by Agrobacterium tumefaciens and a large number of transgenic plants were produced. Embryogenic calli were co-cultivated with A. tumefaciens strain AGL1 carrying the plasmid pCAMBIA1300 containing hygromycin resistance(HygR). Transgenic plants showing in vitro resistance to 50mg/L hygromycin were obtained using a selection procedure. Stable integration of argE and HPT genes into chromosomal DNA was proven by southern blot analysis and PCR analysis of genomic isolated from $T_0$ progenies. The fragments of 650 bp(HPT) were detected in transgenic rice lines. The 230 bp(argE) fragments were showed in agarose gel, and detected fragments were matched with size of argE specific primer. The microscopic feature of leaf on scanning electron microscope(SEM) revealed differences between clear and chalky in shape and arrangement of stoma but did not discriminate.

Agrobacterium tumefaciens에 의해 일미벼 품종에 도입하여 많은 형질전환벼를 생산했다. 배발생 캘러스는 hygromycin 저항성 선발마커를 포함한 pCAMBIA1300 벡터를 이용하여 Agrobacterium strain AGL1으로 공동배양하였다. 공동배양 후 50mg/L hygromycin에 저항성을 보이는 형질전환체가 선발되었다. $T_0$ 세대 형질전환벼로부터 genomic DNA를 추출하여 PCR과 Southern blot을 통한 외래 유전자의 안정한 삽입을 검정하였다. 형질전환된 벼라인에서 650bp의 HPT 유전자 단편이 나타났다. 그리고 argE의 특이적 priomer를 이용하여 검정한 결과 230bp 단편이 검출되었다. SEM을 이용하여 형태학적 분석을 한 결과 기공 분포와 배열상의 특징에 있어서 형질전환체는 대조구와 비교하여 기공형태가 불규칙적인 차이를 보여 주었다.

Keywords

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