Journal of Plant Biotechnology

  • Volume 50
  • /
  • Pages.63-69
  • /
  • 2023
  • /
  • 1229-2818(pISSN)
  • /
  • 2384-1397(eISSN)
The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Agrobacterium-mediated transformation produces transgenic oilseed rape with a high-yield trait

아그로박테리움 형질전환법을 이용한 수량증대 유채 식물체 개발

  • Jong Bo Kim (Department of Biotechnology, College of Biomedical & Health Sciences, Glocal Campus. Konkuk university)
  • 김종보 (건국대학교 글로컬캠퍼스 의료생명대학 생명공학과)
  • Received : 2023.04.22
  • Accepted : 2023.04.24
  • Published : 2023.04.28
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Abstract

This study includes the transformation of genes such as ORE7, the increase of gene expression, and the use of the bar gene as a selectable marker that shows herbicide resistance with Agrobacterium tumefaciens using hypocotyls from the oilseed rape "Youngsan" cultivar. To establish an Agrobacterium transformation system for the production of oilseed rape with a high-yield trait, infection time and co-cultivation period with Agrobacterium were tested. Therefore, when hypocotyls from the oilseed rape "Youngsan" cultivar were infected with Agrobacterium for 20 min and co-cultivated for 3 days, approximately 32-36 putatively transformed hypocotyls with shoots including roots survived from 100 inoculated hypocotyls after 4 weeks of transformation on a selection medium containing 20 mg/L of phosphinothricin (PPT) as a selectable agent. Additionally, a PCR assay was performed to confirm the insertion of target genes and showed the presence of the ORE7 gene as a high-yielding trait and the bar gene as a selectable marker. Treatment with 0.5% (v/v) Basta solution as a selectable agent for 6 days with leaves from transformed oilseed rape expressed the bar gene. Therefore, this study can contribute to the development of special oilseed rapes containing agriculturally useful traits such as herbicide resistance, drought tolerance, high yielding traits, and high oleic acid content.

본 연구는 유채(Brassica napus L.)의 배축을 이용하여 수량 증대 유전자인 ORE7 그리고 선발유전자로 제초제저항성을 나타내는 bar 유전자를 Agrobacterium 기법을 이용하여 형질전환 하였다. 효율적인 유채형질전환 기법을 확립하기 위해 한국 유채 '영산' 품종의 배축 절편체를 이용하여 Agrobacterium 접종 시, 20분간의 접종시간 그리고 3일간의 공동배양기간을 적용할 때 100개의 접종된 배축 절편체들로부터 약 32-36개 개체가 PPT (Phosphinothrixin) 20 mg/l 첨가된 선발배지에서 생존하여 높은 형질전환 효율을 보여주었다. 또한 본 연구에서 도입된 선발 및 생산성 증대 유전자 도입 여부를 확인하기 위해 PCR을 수행하여 도입여부를 확인하였다. 또한 생산성 증대 유전자 ORE 7 유전자와 같이 도입된 bar 유전자의 발현여부를 확인하기 위해 0.5% Basta 용액에 처리한 결과, 제초제저항성 형질이 발현됨을 확인하였다. 본 연구결과를 통해 향후 국내 유채품종을 대상으로 Agrobacterium을 이용하여 제초제 저항성, 건조저항성. 생산성 증대 형질 그리고 오일함량 증대 등의 유용형질 개량에 이용되리라 판단된다.

Keywords

Acknowledgement

이 논문은 건국대학교 2021년 KU 학술연구비 지원에 의한 논문임.

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