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Acta Agron Sin ›› 2011, Vol. 37 ›› Issue (06): 982-990.doi: 10.3724/SP.J.1006.2011.00982

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

GmAREB Gene Improves Tolerances to Drought and Oxidation in Transgenic Arabidopsis

GAO Shi-Qing1,CHEN Ming2,XU Zhao-Shi2,TANG Yi-Miao1,LI Lian-Cheng2,MA You-Zhi2,ZHAO Chang-Ping1   

  1. 1 Beijing Engineering and Technical Research Center for Hybrid Wheat / Beijing Academy of Agricultural and Forestry Science, Beijing 100097, China; 2 National Key Facility for Crop Genetic Resources and Genetic Improvement / Key Laboratory of Crop Genetics and Breeding, Ministry of Agriculture / Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2010-11-12 Revised:2011-03-08 Online:2011-06-12 Published:2011-04-12
  • Contact: 赵昌平, E-mail: cp_zhao@sohu.com; 马有志, E-mail: mayzh@yahoo.com.cn

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Abstract: Abiotic stresses such as drought, high-salt and low temperature severely affect the yield and quality of crops. It is reported that subfamily A of alkalescence leucine zipper transcription factors (basic leucine zipper, bZIP) mainly participated in response to ABA, drought, high salt and oxidative stresses, plays vital roles in stress signal transduction, downstream gene expression regulation and improvement of anti-adversity in plants. In this study, soybean (Glycine max) cv. Tiefeng 8 with salt tolerance was used to isolate and obtain a bZIP transcription factor gene by electronic assembly methods, which was named as GmAREB (Glycine max ABA responsive element binding protein). This gene was composed of 1,317 nucleotides and encoded 439 amino acids. Sequence analysis showed that there were four putative phosphorylation sites (C1, C2, C3, C4), a nuclear localization signal region (KVVE) and a conservative bZIP domain. Homology and phylogenetic trees displayed that GmAREB had the higher homology and more close relationships with Arabidopsis ABF2 and rice TRAB1. To study the combining characteristics of transcription factors and DNA, we carried out electrophoretic mobility shift assay (EMSA) experiment and the results displayed thatGmAREB could specifically bind the ABRE cis-element in vivo. Functional identification of stress treatments showed that the survival rate of 35S::GmAREB transgenic plants treated by drought was higher (50%) than that (5%) of wild type. Stomatal observation displayed that the stomatal aperture of transgenic Arabidopsis (0.8 μm) was less than that of control (2.6 μm) under the drought stress for 2 h. The 35S::GmAREB transgenic plants were treated by methyl viologen (MV) to identify the oxidative stress tolerance of the GmAREB gene. Statistical analysis showed that transgenic Arabidopsis maintained the higher chlorophyl content (7.3 mg g–1 FW) than that of the wild-type. RT-PCR analysis of transgenic Arabidopsis showed that GmAREB overexpression enhanced the expression of downstream stress-related target genes (ABI1, ABI2) and suppressed the stomatal opening-related target genes (KAT1, KAT2). Therefore, the results above illuminated that GmAREB overexpression effectively regulated the expression of downstream target genes, accelerated the stomatal closure, reduced the moisture to evaporate, decreased chlorophyll degradation and enhanced the tolerance to drought and oxidative stresses in transgenic Arabidopsis.

Key words: ABA, bZIP transcription factor, Arabidopsis, Stress tolerance

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