Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Identification of multiple key genes involved in pathogen defense and multi-stress tolerance using microarray and network analysis

Microarray와 Network 분석을 통한 병원균 및 스트레스 저항성 관련 주요 유전자의 대량 발굴

  • Kim, Hyeongmin (Department of Biology, Chungbuk National University) ;
  • Moon, Suyun (Department of Biology, Chungbuk National University) ;
  • Lee, Jinsu (Department of Biology, Chungbuk National University) ;
  • Bae, Wonsil (Department of Biology, Chungbuk National University) ;
  • Won, Kyungho (Pear Research Institute, National Institute of Horticultural & Herbal Science, Rural Development Administration) ;
  • Kim, Yoon-Kyeong (Pear Research Institute, National Institute of Horticultural & Herbal Science, Rural Development Administration) ;
  • Kang, Kwon Kyoo (Department of Horticulture, Hankyong National University) ;
  • Ryu, Hojin (Department of Biology, Chungbuk National University)
  • 김형민 (충북대학교 자연과학대학 생물학과) ;
  • 문수윤 (충북대학교 자연과학대학 생물학과) ;
  • 이진수 (충북대학교 자연과학대학 생물학과) ;
  • 배원실 (충북대학교 자연과학대학 생물학과) ;
  • 원경호 (농촌진흥청 국립원예특작과학원 배연구소) ;
  • 김윤경 (농촌진흥청 국립원예특작과학원 배연구소) ;
  • 강권규 (국립한경대학교 원예학과) ;
  • 류호진 (충북대학교 자연과학대학 생물학과)
  • Received : 2016.08.16
  • Accepted : 2016.08.27
  • Published : 2016.09.30
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Abstract

Brassinosteroid (BR), a plant steroid hormone, plays key roles in numerous growth and developmental processes as well as tolerance to both abiotic and biotic stress. To understand the biological networks involved in BR-mediated signaling pathways and stress tolerance, we performed comparative genome-wide transcriptome analysis of a constitutively activated BR bes1-D mutant with an Agilent Arabidopsis $4{\times}44K$ oligo chip. As a result, we newly identified 1,091 (562 up-regulated and 529 down-regulated) significant differentially expressed genes (DEGs). The combination of GO enrichment and protein network analysis revealed that stress-related processes, such as metabolism, development, abiotic/biotic stress, immunity, and defense, were critically linked to BR signaling pathways. Among the identified gene sets, we confirmed more than a 6-fold up-regulation of NB-ARC and FLS2 in bes1-D plants. However, some genes, including TIR1, TSA1 and OCP3, were down-regulated. Consistently, BR-activated plants showed higher tolerance to drought stress and pathogen infection compared to wild-type controls. In this study, we newly developed a useful, comprehensive method for large-scale identification of critical network and gene sets with global transcriptome analysis using a microarray. This study also showed that gain of function in the bes1-D gene can regulate the adaptive response of plants to various stressful conditions.

브라시노스테로이드는 식물의 생장과 발육 과정에 있어서 중요한 역할을 담당 할 뿐 아니라 생물학적/ 비 생물학적 스트레스에 대한 복합 저항성을 보인다고 알려져 있다. 따라서 본 연구에서는 브라시노스테로이드와 광범위스트레스 내성을 연결하는 중요한 생물학적 네트워크를 이해하기 위해, Agilent Arabidopsis $4{\times}44K$ oligo chip을 이용하여 브라시노스테로이드 신호가 강화된 bes1-D 계통의 전 전사체 비교분석을 수행하였다. 그 결과 bes1-D 계통에서 DEGs (Differentially Expressed Genes)를 1,091 (562 up-regulated, 529 down-regulated) 개 선발하였다. 또한 선발된 유전자들의 GO 와 단백질 상호작용 네트워크 분석을 통해 대사, 발달, 스트레스, 면역, 방어 반응에 관련된 주요 브라시노스테로이드 신호전달과 연결된 스트레스 관련 유전자군을 분리하였다. 선발된 유전자중 NB-ARC와 FLS2는 bes1-D 계통이 야생형 En-2 계통에 비해 약 6배 정도의 발현량이 증가되었으며, TIR1, TSA1, OCP3 유전자등은 bes1-D 계통이 야생형 En-2 계통에 비해 발현이 감소되었다. 또한 브라시노스테로이드 활성형 계통이 야생형 식물체 계통에 비해 가뭄 스트레스 및 병원균에 대해 저항력이 향상되었다. 따라서 microarray 분석을 통한 유전자 간 발현 네트워크와 유전체 정보를 결합하여 대단위 주요 기능 유전자들을 동정할 수 있는 방법을 고안하여 실험에 사용하였다. 이를 통해 기능 획득 돌연변이 bes1-D가 식물들이 다양한 스트레스 환경에 적응할 수 있는 반응을 조절한다는 사실을 보여주고 있다.

Keywords

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