Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Functional Analysis of a Grapevine UDP-Glucose Flavonoid Glucosyl Transferase (UFGT) Gene in Transgenic Tobacco Plants

담배 형질전환체를 이용한 포도 UDP-glucose flavonoid glucosyl transferase (UFGT) 유전자의 기능 분석

  • Park, Ji-Yeon (Department of Molecular Biology, BK21 Graduate Program for RNA Biology and Institute of Nanosensor and Biotechnology, Dankook University) ;
  • Park, Sung-Chool (Department of Molecular Biology, BK21 Graduate Program for RNA Biology and Institute of Nanosensor and Biotechnology, Dankook University) ;
  • Pyee, Jae-Ho (Department of Molecular Biology, BK21 Graduate Program for RNA Biology and Institute of Nanosensor and Biotechnology, Dankook University)
  • 박지연 (단국대학교 분자생물학과, BK21 RNA 전문인력양성팀, 나노센서바이오텍연구소) ;
  • 박성출 (단국대학교 분자생물학과, BK21 RNA 전문인력양성팀, 나노센서바이오텍연구소) ;
  • 피재호 (단국대학교 분자생물학과, BK21 RNA 전문인력양성팀, 나노센서바이오텍연구소)
  • Received : 2009.11.11
  • Accepted : 2010.02.08
  • Published : 2010.02.28
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Abstract

Anthocyanin, a phenolic compound, is a pigment that shows blue or red color in the fruit, petal and other tissues. It is an important factor in grape berry skin pigment and accumulates only in the skin. This skin-specific accumulation of anthocyanin has been reported to be regulated by the ufgt gene which encodes UDP-glucose: flavonoid 3-O-glucosyltransferase that participates in the biosynthesis of anthocyanin. The ufgt gene is expressed only in berry skin, while the other genes involved in the biosynthetic pathway are expressed in both skin and flesh tissues. In order to determine whether anthocyanin accumulation is primarily regulated by compartment of UFGT, a ufgt cDNA clone was isolated from grape berry, its open reading frame was ligated in pBI121 vector in either a sense or an antisense orientation under the control of the CaMV35S promoter and the recombinant constructs were incorporated into tobacco plants. Several transgenic lines were selected and characterized to determine the level of expression of the grapevine ufgt transcript and endogenous homologs of tobacco. Compared to the wild-type, the amount of anthocyanins in sense transgenic plants increased by 44%, while the amount of anthocyanins in antisense transgenic plants decreased by 88%. In addition, the color of flowers became intense in the sense transgenic plants. These results suggest that over-expression or repression of the ufgt gene affected the accumulation of anthocyanin in flowers of tobacco.

페놀화합물인 안토시아닌은 과일, 꽃잎 등 식물 조직에서 청 혹은 적색을 나타내는 색소이다. 포도의 경우, 안토시아닌은 적포도 품종의 과피에만 축적되는데, 이것은 안토시아닌 생합성에 관여하는 효소 중에서 UDP-glucose: flavonoid 3-O-glucosyltransferase(UFGT)를 암호화하는 ufgt 유전자에 의해 조절된다고 보고된 바 있다. ufgt 유전자에 의해 안토시아닌의 축적 양상이 변하는지를 검증하기 위해, 포도로부터 ufgt cDNA를 분리한 다음, 각각 sense와 antisense 방향으로 발현하는 벡터를 제작하고, 이를 야생형 담배에 도입하였다. 담배 형질전환체를 선발하여 RT-PCR을 수행한 결과, 포도 ufgt 유전자가 sense 방향으로 들어간 snese 형질전환체에서는 야생형 담배에 비해 ufgt 전사체의 양이 증가하였고, antisense 방향으로 들어간 antisense 형질전환체에서는 전사체의 양이 도리어 감소하였다. 한편, 담배 형질전환체의 꽃을 분석한 결과, sense 형질전환체의 안토시아닌 함량은 야생형 담배에 비해 최고 44% 증가하였고, antisense 형질전환체에서는 최고 88% 감소하였다. 또한 sense 형질전환체의 꽃 색깔은 야생형 담배에 비해 더 진해졌다. 이러한 결과는 외부로부터 도입된 포도 ufgt 유전자의 발현으로 ufgt 전사체의 양이 증가하면 담배 내 안토시아닌의 함량이 높아지고 꽃 색깔이 진해진다는 것을 시사한다.

Keywords

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