비 생물학적 스트레스 시 벼에서 OsABF3 유전자 분리와 ABA 신호전달 대한 연구
Studies on OsABF3 Gene Isolation and ABA Signal Transduction in Rice Plants Against Abiotic Stress
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ABA는 식물에서 비 생물학적 스트레스 내성에 관여하는 중요한 식물 호르몬이다. 애기장대의 group A bZIP 전사인자는ABA 신호전달 과정에 중요한 역할을 한다고 알려져 있다. 그러나 벼에서는 group A bZIP 전사인자의 기능이 잘 알려져 있지않다. 따라서 우리는 벼에서 group A bZIP 전사인자인 OsABF3 (Oryza sativa ABA responsive element binding factor 3)를 연구하였다. 이를 위해 벼의 다양한 조직과 다양한 스트레스(가뭄, 염분, 저온, ABA, 산화 스트레스)에 따른 OsABF3 발현 패턴을 분석하였다. 또한 maize의 원형질체에서 GFP fusion 벡터를사용한 세포 내 위치 분석을 통해 OsABF3가 핵단백질이라는 것을 확인하였다. Yeast one-hybrid 실험을 통해 OsABF3의 Cterminal 부분이 ABREs에 결합한다는 것과 N-terminal 부분이 하위 유전자의 transactivation 하는데 필요하다는 것을 알수 있었다. 그리고 T-DNA가 삽입된 OsABF3의 homozygous 돌연변이체가 야생형과 과발현체에 비해 발아와 발아 후 단계에서 고농도의 ABA에 대한 민감도가 더 감소한 것을 알 수 있었다. 결과적으로 종합해 볼 때 OsABF3는 ABA의 의존적인경로를 통해 비 생물학적 스트레스에 반응하는 유전자의 발현을 조절하는 기능을 하는 전사 조절자이다. 또한 OsABF3의transactivation을 측정하는 실험에 있어서 억제 domain이 존재한다는 결과를 얻었다.
Abstract
Abscisic acid (ABA) is an important phytohormone involved in abiotic stress tolerance in plants. The group A bZIP transcription factors play important roles in the ABA signaling pathway in Arabidopsis but little is known about their functions in rice. In our current study, we have isolated and characterized a group A bZIP transcription factor in rice, OsABF3 (Oryza sativa ABA responsive element binding factor 3). We examined the expression patterns of OsABF3 in various tissues and time course analysis after abiotic stress treatments such as drought, salinity, cold, oxidative stress, and ABA in rice. Subcellular localization analysis in maize protoplasts using a GFP fusion vector further indicated that OsABF3 is a nuclear protein. Moreover, in a yeast one-hybrid experiment, OsABF3 was shown to bind to ABA responsive elements (ABREs) and its N-terminal region found to be necessary to transactivate a downstream reporter. A homozygous T-DNA insertional mutant of OsABF3 is more sensitive to salinity, drought, and oxidative stress compared with wild type plants ＆ OsABF3OX plants. In addition, this Osabf3 mutant showed a significantly decreased sensitivity to high levels of ABA at germination and post-germination. Collectively, our present results indicate that OsABF3 functions as a transcriptional regulator that modulates the expression of abiotic stress- responsive genes through an ABA-dependent pathway.
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