Korean Journal of Plant Resources (한국자원식물학회지)

The Plant Resources Society of Korea (한국자원식물학회)
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An Effective Selection of PAT Gene Transformed Populus alba $\{times}$ Populus glandulosa No.3 using Herbicide Basta Treatment

제초제 Basta를 이용한 Phosphinothricin Acetyltransferase 유전자로 형질전환된 현사시 3호의 효율적인 선발

  • 오경은 (충북대학교 산림과학부, 임업연구원 임목육종부) ;
  • 문흥규 (충북대학교 산림과학부, 임업연구원 임목육종부) ;
  • 박재인 (경희대학교 생명과학대학 및 한방재료가공연구센타) ;
  • 양덕춘 (경희대학교 생명과학대학 및 한방재료가공연구센타)
  • Published : 2004.02.01
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Abstract

This study was conducted to simple transformants selection by herbicide Basta treatment after transformation with Agrobacteium tumefaciens MP90/PAT in hybrid poplar(Populus alba ${\times}$ P. glandulosa No. 3). In preliminary study, we determined that the lethal concentration of herbicide Basta was 1.0mg/L in callus culture, adventitious bud formation and axillary bud elongation experiment. By the treatment of 1.0mg/L Basta, we could be selected the transformed shoots effectively from the various cultures. In addition, the treatment was useful on selection of transformants which are growing in soil pot. Finally, we also confirmed the transformation by PAT assay, Above results show that the herbicide Basta treatment and PAT assay can be a very simple and effective method for the identification of PAT gene transformed hybrid poplar.

본 연구는 개량 포플러 현사시 3호를 재료로 제초제 저항성 유전자 PAT으로 형질전환시 킨 다음 제초제 Basta처리에 의한 효율적인 형질전환체 선발을 목적으로 시험되었다. Basta 처리에 따른 캘러스유도, 부정아유도 및 액아유도 시험을 통해 조직치사 농도는 1.0mg/L로 결정할 수 있었으며, 이 농도처리에 의해 형질전환체의 조기선발에 사용하였다. 이 농도처리로 비형질전환체는 모두 고사되었지만, 형질전환체는 정상적으로 반응을 보이며, 생육이 가능하였다 .한편, 형질전환체에 제초제를 직접 살포한 결과, 정상적 생육이 가능했고, PAT activity측정 결과에서도 양성 반응을 나타내어 현사시 3호 세포내에 PAT유전자가 정상 발현됨이 확인되었다. 이로써 현사시에 PAT유전자 도입 및 발현을 위하여 제초제 Basta 처 리와 PAT assay가 매우 효과적인 것으로 나타났다.

Keywords

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