Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Transgenic Tomato Plants That Overexpress Superoxide Dismutase in Fruits

토마토 과실에서 Superoxide Dismutase를 고발현하는 형질전환 식물체

  • Park, Eun-Jeong (Plant Cell Biotechnology Laboratory, Korea Research Institute of Bioscience and Biotechnology) ;
  • Lee, Haeng-Soon (Plant Cell Biotechnology Laboratory, Korea Research Institute of Bioscience and Biotechnology) ;
  • Kwon, Suk-Yoon (Plant Cell Biotechnology Laboratory, Korea Research Institute of Bioscience and Biotechnology) ;
  • Choi, Kwan-Sam (Department of Agricultural Biology, Chungnam National University) ;
  • Kwak, Sang-Soo (Plant Cell Biotechnology Laboratory, Korea Research Institute of Bioscience and Biotechnology)
  • 박은정 (한국생명공학연구원 식물세포공학연구실) ;
  • 이행순 (한국생명공학연구원 식물세포공학연구실) ;
  • 권석윤 (한국생명공학연구원 식물세포공학연구실) ;
  • 최관삼 (충남대학교 농생물학과) ;
  • 곽상수 (한국생명공학연구원 식물세포공학연구실)
  • Published : 2002.03.01
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Abstract

Superoxide dismutase (SOD) plays an important role in cellular defense against oxidative stress in plants. We have developed transgenic tomato plants overexpressing a cassava SOD in fruits. Three transgenic tomato plants (one from cv. Pink forcer and two from cv. Koko) using a new vector system, ASOp :: . mSOD1/pBI101, harboring ascorbate oxidase promoter (ASOp) expressing dominantly in cucumber fruits, CuZnSOD cDNA (mSOD1) isolated from cultured cells of cassava, and nptll gene as a selectable marker were successfully developed. SOD specific activity (units/mg protein) in transgenic fruits of both cultivars was increased with maturation of the fruits. SOD specific activity of well-mature fruits in transgenic Pink forcer and Koko showed approximately 1.6 and 2.2 times higher than control fruits, respectively. The strength of SOD isoenzyme bands well reflected the SOD activity during the fruit maturation. These results suggested that SOD gene was properly introduced into tomato fruits in a fruit-dominant expression manner by ASO promoter.

Superoxide dismutase (SOD)를 과실에서 고발현시킨 형질전환 토마토 (서광과 꼬꼬)를 개발하였다. 카사바 배양세포에서 분리한 CuZnSOD (mSOD1)를 과실에 우세적으로 발현하는 ascorbate oxidase promoter (ASOp)를 이용하여 ASOp :: mSOD1/pBI101 벡터를 제작한 후 Agrobacterium 매개로 자엽 절편체를 형질전환하였다. Kanamycin 저항성 식물체를 기관발생 경로로 재분화시킨 후 Southern 분석으로 형질전환을 확인하였다. 서광과 꼬꼬 토마토의 형질전환체와 대조구 식물체의 과실을 성숙 단계별로 분류하여 단백질 함량과 SOD 비활성도 (units/mg protein)를 측정한 결과, 단백질 함량은 열매가 익은 단계로 갈수록 점점 감소하여 완전히 익은 단계에서 가장 낮았다. SOD 비활성도는 형질전환 토마토의 열매의 모든 단계에서 대조구보다 높았으며 완전히 성숙한 과실에서 가장 높았다. 성숙한 형질전환 서광과 꼬꼬 과실에서 SOD 비활성도는 비형질전환의 것보다 각각 약 1.6배와 약 2.2배 높았다. SOD isoenzyme gel 분석에서 도입한 mSOD1로 추정되는 CuZnSOD 밴드가 형질전환체에서 과실 성숙에 따라 강하게 발현되었다. 이상의 결과로서 ASO promoter에 의해 SOD 유전자가 토마토 과실에 특이적으로 발현됨이 확인되었다.

Keywords

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