Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Isolation and characterization of a monodehydroascorbate reductase gene in poplar (Populus alba × P. glandulosa)

현사시나무 monodehydroascorbate reductase (MDHAR) 유전자의 분리 및 발현특성

  • Yoon, Seo-Kyung (Division of Forest Biotechnology, Korea Forest Research Institute) ;
  • Park, Eung-Jun (Division of Forest Biotechnology, Korea Forest Research Institute) ;
  • Bae, Eun-Kyung (Division of Forest Tree Improvement, Korea Forest Research Institute) ;
  • Choi, Young-Im (Division of Forest Biotechnology, Korea Forest Research Institute) ;
  • Kim, Joon-Hyeok (Division of Forest Biotechnology, Korea Forest Research Institute) ;
  • Lee, Hyoshin (Division of Forest Tree Improvement, Korea Forest Research Institute)
  • 윤서경 (국립산림과학원 산림생명공학과) ;
  • 박응준 (국립산림과학원 산림생명공학과) ;
  • 배은경 (국립산림과학원 임목육종과) ;
  • 최영임 (국립산림과학원 산림생명공학과) ;
  • 김준혁 (국립산림과학원 산림생명공학과) ;
  • 이효신 (국립산림과학원 임목육종과)
  • Received : 2014.09.19
  • Accepted : 2014.10.20
  • Published : 2014.12.31
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Abstract

Monodehydroascorbate reductase (MDHAR) is an important enzyme that plays a role in the detoxification of reactive oxygen species (ROS) by maintaining reduced pool of ascorbate through recycling the oxidized form of ascorbate. In this study, we isolated a PagMDHAR1 gene from Populus alba ${\times}$ P. glandulosa, and investigated its expression characteristics. The PagMDHAR1 cDNA encodes a putative 434 amino acids containing FAD- and NAD(P)H-binding domains. Southern blot analysis indicated that a single nuclear gene encodes this enzyme. Northern hybridization analysis revealed that PagMDHAR1 is highly expressed in both suspension cells and flower tissues, while its expression levels were enhanced by drought, salt, cold, wounding and ABA. Therefore, PagMDHAR1 might be expressed in response to abiotic stress through the ABA-mediated signaling pathway in this poplar species, suggesting that the PagMDHAR1 plays an important role in the defense mechanisms against oxidative stress.

Monodehydroascorbate reductase (MDHAR)는 활성산소종 제거에 중요한 효소이다. 본 연구에서는 MDHAR 유전자를 현사시나무(Populus alba ${\times}$ P. glandulosa)에서 분리하여 이를 PagMDHAR1이라 명명하고 유전자의 구조와 발현특성을 조사 하였다. PagMDHAR1 유전자는 434개의 아미노산으로 구성된 단백질을 암호화하며 3개의 FAD/NAD(P)H 결합 영역이 보존되어 있다. PagMDHAR1은 현사시나무의 염색체에 1 ~ 2 copy가 존재하며, 배양세포와 꽃에서 높게 발현하였다. 현탁배양세포의 생장주기에서는 유도기와 초기 지수생장기에서 높게 발현하였다. 또한 PagMDHAR1은 건조와 염, 저온, 상처 및 ABA 처리에 의해서 발현이 증가하는 것으로 나타났다. 따라서 PagMDHAR1은 ABA를 경유한 신호전달경로를 따라 다양한 스트레스에 반응하며, PagMDHAR1의 기능이 활성산소종에 의해 유도되는 산화 스트레스 방어기작에서 중요한 역할을 할 뿐만 아니라 스트레스 내성에도 기여할 것으로 생각된다. 이는 향후 PagMDHAR1 형질전환 식물체 생산 등 생명공학적 기술을 이용한 유전자 기능에 대한 연구를 비롯하여 신기능성 임목의 개발에 활용 가능할 것으로 기대된다.

Keywords

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