Horticultural Science & Technology (원예과학기술지)

Korean Society of Horticultural Science (한국원예학회)
권호 표지

Enhanced Antioxident Effect by over Expression of Tomato β-carotene Hydroxylase Gene (ChyB) Using Agrobacterium-infiltration in Tobacco Plant

Agro-infiltration을 이용한 토마토 β-carotene hydroxylase 유전자(ChyB) 과발현 및 담배식물체의 항산화 효과 증진

  • Choi, Yoon-Jeong (Department of Horticulture and Life Science, Yeungnam University) ;
  • Yoon, Kyung-Young (Department of Food and Nutrition, Yeungnam University) ;
  • Yun, Hae-Keun (Department of Horticulture and Life Science, Yeungnam University) ;
  • Suh, Sang-Gon (Department of Horticulture and Life Science, Yeungnam University) ;
  • Moon, Yong-Sun (Department of Horticulture and Life Science, Yeungnam University)
  • 최윤정 (영남대학교 원예생명과학과) ;
  • 윤경영 (영남대학교 식품영양학과) ;
  • 윤해근 (영남대학교 원예생명과학과) ;
  • 서상곤 (영남대학교 원예생명과학과) ;
  • 문용선 (영남대학교 원예생명과학과)
  • Received : 2011.03.15
  • Accepted : 2011.04.22
  • Published : 2011.06.30
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Abstract

Several reports indicated that astaxanthin and zeaxanthin have more active anticancer activity than pro-vitamin A carotenes. ${\beta}$-carotene hydroxylase is a key enzyme to synthesize zeaxanthin and astaxanthin in carotenoids biosynthesis pathway. We isolated the ChyB gene encoding ${\beta}$-carotene hydroxylase from tomato leaves. The ChyB gene (1.5Kbp) fragment was cloned into the binary vector and designated to pIG121-ChyB-tom. Agrobacterium-mediated infiltration was used for transient assay in Nicotiana benthamiana. Leaf samples were collected 0, 1, 2, 3 days after infiltration (DAI). RT-PCR result showed that the expression of ${\beta}$-carotene hydroxylase transcripts was not detected in control (0DAI), but its expression was detected after 1 DPI and increased later on. When the activity of ${\beta}$-carotene hydroxylase was measured, the 1,1-diphenyl-pricryl hydrazyl (DPPH) radical scavenging activity (27%) at 2 DAI was significantly higher than that (21%) at 0 DAI. These results indicated that anti-oxidant activity dramatically increased at 2 DAI in tobacco leaves was due to over expression of tomato ${\beta}$-carotene hydroxylase. These results can be the foundation to develop tomato cultivars with high oxy-carotenoids content using the ChyB gene transformation.

${\beta}$-carotene hydroxylase는 carotenoids의 생합성 경로에서 astaxanthin 및 zeaxanthin 합성에 관여하는 주요 효소이다. Astaxanthin과 zeaxanthin은 비타민 A 전구체보다 항암효과가 더 높다고 보고되고 있다. 따라서 ${\beta}$-carotene hydroxylase 유전자(ChyB)를 토마토 잎에서 분리하고 binary 벡터에 클로닝 한 후 pIG121-ChyB-tom으로 명명하였다. 토마토 ChyB 유전자를 Agrobacterium-mediated infiltration 방법을 이용하여 생육 8주 된 담배 잎에 일시적 형질전환을 하였다. 감염 0, 1, 2, 3일 후 RT-PCR 한 결과 담배에서는 거의 발현되지 않았던 ${\beta}$-carotene hydroxylase 전사체가 감염 1일 후부터 증가하여 2일 후 최대 전사량을 보이고 3일 후부터 감소하는 경향을 나타내었다. 이 결과를 바탕으로 감염 후 0, 2, 3일째 담배 잎을 채취하여 항산화 효능을 측정한 결과, 감염 2일 후 1,1-diphenyl-pricryl hydrazyl(DPPH) radical 소거 활성이 대조구에 비해 약 30% 증가하는 것을 확인하였다. 이 연구결과를 통해 외래 유용 유전자를 원예작물에 형질전환하여 항산화 및 항암효과가 우수한 zeaxanthin 및 astaxanthin 등 oxy-carotenoids의 함유량을 증진시킨 새로운 기능성 토마토 및 다양한 원예작물의 신품종 개발에 이용될 수 있을 것으로 기대된다.

Keywords

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