Korean Journal of Environment and Ecology (한국환경생태학회지)

Korean Society of Environment and Ecology (한국환경생태학회)
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Molecular Isolation and Characterization of the 2CysPrx Gene from Salicornia herbacea

퉁퉁마디로부터 2CysPrx 유전자 분리 및 특성 분석

  • Kim, Suk-Kyu (Tidal Flat Research Center-National Institute of Fisheries Science) ;
  • Chung, Sang Ok (Tidal Flat Research Center-National Institute of Fisheries Science) ;
  • Na, Gil-Jong (Dept. of Biology, Kunsan Nat'l Univ.)
  • 김석규 (국립수산과학원 서해수산연구소 갯벌연구센터) ;
  • 정상옥 (국립수산과학원 서해수산연구소 갯벌연구센터) ;
  • 나종길 (군산대학교 생물학과)
  • Received : 2016.08.17
  • Accepted : 2016.09.29
  • Published : 2016.10.30
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Abstract

This study is focused on the investigation of the genes which are induced by various stresses of the halophyte Salicornia herbacea. One of the factors influencing in the germination of Salicornia herbacea is salt stress. The highest germination rate was found in the condition without NaCl, and the upper limit of the NaCl concentration for the germination of Salicornia herbacea was 7%. The optimal temperature of $20^{\circ}C$showed a germination rate of 98%. Among genes induced by stress the 2CysPrx gene was cloned and analyzed for this study. The 2CysPrx gene has two cysteine conserved residues and is composed of 275 amino acids with molecular weight of 30.1kDa. The 2CysPrx gene appeared to be one copy in the genome and consists of 6 introns and 7 exons. Quantitative real-time PCR revealed that the highest transcription rate induced by NaCl and $H_2O_2$ appeared to be at the concentration of 3.5% NaCl and 40mM $H_2O_2$, respectively. The amount of transcript induced by high temperature($40^{\circ}C$) and $75{\mu}M$ of ABA was respectively highest. The gene at low temperature ($4^{\circ}C$) appeared not to be expressed. We are conducting to clone other peroxyredoxin genes induced by various environmental stresses.

염생식물 퉁퉁마디의 종자 발아에 영향을 미치는 환경 요인을 조사하고 환경 스트레스에 의해 유도되는 2CysPrx 유전자를 클로닝한 후 스트레스 조건에 따른 2CysPrx 유전자의 발현 양상에 대하여 조사하였다. 염생식물에 대한 가장 대표적인 스트레스는 염분 스트레스로서 퉁퉁마디 발아에 중요한 요인으로 작용하고 있다. 퉁퉁마디의 발아에 대한 NaCl의 한계 농도는 7%로 나타났고, 최적의 발아 조건은 NaCl이 없는 상태로 확인되었다. 퉁퉁마디 발아에서 최적 온도는 $20^{\circ}C$로 98%의 발아율을 보였다. 스트레스에 유도되는 유전자 후보군 중 2CysPrx 유전자의 cDNA를 클론하여 분석한 결과 275개의 아미노산으로 이루어져 있고 두 개의 시스테인 잔기를 가지고 있으며 분자량은 30.1kDa으로 나타났다. 2CysPrx 유전자는 서던 블롯에 의해 유전체에 한 카피 존재하는 것으로 나타났고, 6개의 인트론과 7개의 엑손으로 구성되어 있다. qPCR에 의한 2CysPrx 유전자의 전사율을 분석한 결과, 3.5% NaCl과 40mM $H_2O_2$ 처리 조건에서 전사율이 가장 높게 나타났고, 고온($40^{\circ}C$)과 $75{\mu}M$ ABA 처리 조건에서는 처리 후 8시간에 최고의 전사율을 보였으며, 저온($4^{\circ}C$)에서는 유전자 발현이 일어나지 않는 것으로 나타났다. 우리는 여러 환경 스트레스에 의해 유도되는 다른 유전자의 클로닝을 시도하고 있다.

Keywords

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