Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Selection of transgenic sweetpotato plants expressing 2-Cys peroxiredoxin with enhanced tolerance to oxidative stress

Peroxiredoxin 유전자 발현 산화스트레스 내성 형질전환 고구마의 선발

  • Kim, Myoung-Duck (Environmental Biotechnology Research Center Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Yang, Kyoung-Sil (Environmental Biotechnology Research Center Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Kwon, Suk-Yoon (Plant Systems Engineering Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Lee, Sang-Yeol (Division of Applied Life Science, EB-NCRC and Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University) ;
  • Kwak, Sang-Soo (Environmental Biotechnology Research Center Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Lee, Haeng-Soon (Environmental Biotechnology Research Center Korea Research Institute of Bioscience and Biotechnology (KRIBB))
  • 김명덕 (한국생명공학연구원 환경바이오연구센터) ;
  • 양경실 (한국생명공학연구원 환경바이오연구센터) ;
  • 권석윤 (한국생명공학연구원 식물시스템공학연구센터) ;
  • 이상열 (경상대학교 국가핵심연구센터) ;
  • 곽상수 (한국생명공학연구원 환경바이오연구센터) ;
  • 이행순 (한국생명공학연구원 환경바이오연구센터)
  • Published : 2009.03.31
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Abstract

In order to develop transgenic sweetpotato plants [Ipomoea batatas (L.) Lam. cv. Yulmi] with enhanced tolerance to oxidative stress, we constructed transformation vectors expressing 2-Cys peroxiredoxin (Prx) gene under the control of the stress-inducible SWPA2 or enhanced 35S promoter (named as SP or EP). Transgenic sweetpotato plants were attempted to generate from embryogenic calli using an Agrobacterium-mediated transformation system. Embryogenic calli gave rise to somatic embryos and then converted into plantlets on MS medium containing 100 mg/L kanamycin. Transgenic plants were regenerated in the same medium. Southern blot analysis confirmed that the Prx gene was inserted into the genome of the plants. To further study we selected the transgenic plant lines with enhanced tolerance against methyl viologen (MV). When sweetpotato leaf discs were subjected to methyl MV at $20{\mu}M$, transgenic plants showed about 40% higher tolerance than non-transgenic or empty vector-transformed plants.

산화스트레스에 내성을 지닌 형질전철 고구마 식물체를 개발하기 위해서 산화스트레스에 의해 발현이 강하게 유도되는 SWPA2 프로모터 또는 CaMV 35S 프로모터에 2-Cys peroxiredoxin (Prx) 유전자가 발현되도록 연결한 형질전철 벡터 (pSP-K, pEP-K)를 제작한 후, 각각 Agrobacterium 매개로 형질전환 하였다. 카나마이신 저항성 배발생 캘러스로부터 체세포배발생 과정을 거쳐 100mg/L kanainycin이 포함된 MS 배지에서 소식물체로 발달하였다. Southern 분석으로 외래 유전자가 안정적으로 고구마 게놈 내로 삽입되었음을 확인하였다. 형질전환 고구마 잎 조직을 대상으로 $20{\mu}M$ methyl viologen에 대한 내성 검정을 조사하여 형질전환 고구마 식물체가 비형질전환 식물체 또는 벡터 대조구 식물체 보다 40% 정도 높은 신화스트레스에 대한 내성을 보여주었다. 선발된 형질전환 식물계는 저온, 건조 등의 여러 기지 환경스트레스 내성검정에 이용될 것이며 향후 복합재해 내성 고구마 계통육성에 이용될 수 있을 것으로 기대된다.

Keywords

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