Journal of The Korean Society of Grassland and Forage Science (한국초지조사료학회지)

The Korean Society of Grassland and Forage Science (한국초지조사료학회)
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Production of Transgenic Orchardgrass Overexpressing a Thermotolerant Gene, DgP23

내열성 유전자 DgP23을 도입한 형질전환 오차드그라스의 생산

  • Kim Ki-Yong (National Livestock Research Institute) ;
  • Jang Yo-Soon (Korea Ocean Research & Development Institute) ;
  • Park Geun Je (National Livestock Research Institute) ;
  • Choi Gi Jun (National Livestock Research Institute) ;
  • Seong Byung Ryul (National Livestock Research Institute) ;
  • Seo Sung (National Livestock Research Institute) ;
  • Cha Joon-Yung (Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University) ;
  • Son Daeyong (Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University)
  • Published : 2005.12.01
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Abstract

To develop transgenic orchardgrass (Dactylis glomerata L.) resistant to high temperature, a thermptolerance gene, DgP23, was introduced into orchardgrass using Agrobacterium - mediated transformation method. PCR and Southern blot analyses using genomic DNA showed specific DNA band on agarose gel and hybridization signal on X- ray film in transgenic orchardgrass harboring the recombinant DgP23 gene, but not in the wild type and empty vector control plants. RT-PCR and Southern blot analyses using total RNA also showed specific DNA band and hybridization signal. Transgenic orchardgrass did not showed ny morphological aberration both in the green house and field cultivation. Thermotolerance of transgenic plants was not detected in laboratory test. but may detected in field test.

고온 내성 오차드그라스를 개발하기 위하여, 재조합 DgP23 유전자에 CaMV 35S 프로모터를 붙여 발현벡터를 제작, Agrobacterium 형질전환 방법으로 오차드그라스 형질전환체를 생산하였다. Genomic DNA를 분리하여 PCR 및 Southern blot 분석을 실시한 결과, PCR 분석에서 DgP23 유전자의 DNA band가 관찰되었고, Southern blot 분석에서도 X-ray film 상에 hybridization signal이 관찰되어, 오차드그라스 genome에 DgP23 유전자의 도입이 확인되었으며, wild type 및 empty vector control에서는 DNA band 및 hybridization signal이 관찰되지 않았다. 또한 RT-PCR 및 이들 산물의 Southern blot 분석 결과, DgP23 유전자의 정상적인 발현이 확인되었다. 형질전환 오차드그라스를 온실 및 포장에서 재배하며 생육특성을 조사한 결과, 비형질전환체와 비교하여 형태적 차이는 나타내지 않았다. 실험실 조건에서 고온내성을 조사한 결과, 고온내성이 확인되지 않았기 때문에 형질전환 종자를 생산하여 포장조건에서 고온내성을 검정할 계획이며, 재배시험에서는 내성이 강한 개체를 선발할 수 있을 것으로 예상된다.

Keywords

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