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Increased Resistance of Arabidopsis to Cold and Salt Stresses by Suppresing the Transcription Repressors of the A-5 Group among the DREB Subfamily Transcription Factors through Artificial microRNA |
ZHOU Lu, DONG Chun-juan, LIU Jin-yuan |
MOE Laboratory of Protein Science, School of Life Science, Tsinghua University, Beijing 100084, China |
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Abstract Transcription repressors are proteins that bind to specific sites on DNA and prevent transcription of nearby genes through interacting with transcripitonal activators, acting on the basal transcription factor complex or chromatin remodeling. Among the DREB subfamily, there are 6 transcription repressors containing the EAR motif in the A-5 group, functional analysis of some repressor indictaed it could modulate plant response to stress, however their specific functions and mechanisms are reqiured further research. By designing the amiRNA-A5, which could specifically target the mRNAs of these transcription repressors, and transforming the pCAMBIA-pre-amiRNA-A5 expression vector to Arabiopsis, 9 lines of T2 transgenetic homozygotes were got, selected by Hygromycin and PCR. Real-time PCR assays demonstrated that compared to the wild type plants, the relative expression levels of target genes in transgenetic plants were obviously decreased. To analyze the resistance of these transgenetic plants under cold and salt stresses, relative electrolyte leakage (REL) and malondialdehyde (MDA) content were measured. Both of these two indexes were increased less in the transgenetic plants than in the wilde type plants under stress conditions, suggesting the increased resistance of the transgenetic plants to cold and salt stress. The results indicated these transcription repressors might negatively function under cold and salt stress, provding clues for further functional mechanism study of the transciption repressors of the A-5 group among the DREB subfamily.
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Received: 23 December 2010
Published: 27 May 2011
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[1] Shinozaki K, Yamaguchi-Shinozaki K. Molecular responses to dehydration and low temperature: differences and cross-talk between two stress signaling pathways. Curr Opin Plant Biol,2000,3(3):217-223.
[2] Sakuma Y,Liu Q,Dubouzet J G,et al. DNA-binding specificity of the ERF/AP2 domain of Arabidopsis DREBs, transcription factors involved in dehydration and cold inducible gene expression. Biochem Biophys Res Commun,2002,290(3):998-1009.
[3] Okamuro J K,Caster B,Villarroel R,et al. The AP2 domain of the APETALA2 defines a large new family of DNA binding proteins in Arabidopsis. Proc Natl Acad Sci USA,1997,94(13):7076-7081.
[4] Dong C J,Liu J Y. The Arabidopsis EAR-motif-containing protein RAP2.1 functions as an active transcriptional repressor to keep stress responses under tight control. BMC Plant Biol,2010,10:47.
[5] 叶梅霞,刘军梅,李昊,等. amiRNAi实现高效稳定的特异基因沉默新方法. 中国生物工程杂志,2010,30(8):118-125. Ye M X,Liu J M,Li H,et al. China Biotechnology,2010,30(8) : 118-125.
[6] Niu Q W,Lin S S,Reyes J,et al. Expression of artificial microRNAs in transgenic Arabidopsis thaliana confers virus resistance. Nature Biotech,2006,24(11):1420-1428.
[7] Han P, Li Q, Zhu Y X. Mutation of Arabidopsis BARD1 causes meristem defects by failing to confine WUSCHEL expression to the organizing center. Plant Cell,2008,20(6):1482-1493.
[8] Clough S J,Bent A F. Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant Journal,1998,16(6):735–743.
[9] Huang J,Sun S J,Xu D Q,et al. Increased tolerance of rice to cold, drought and oxidative stresses mediated by the overexpression of a gene that encodes the zinc finger protein ZFP245. Biochem Biophys Res Commun,2009,389(3):556-561.
[10] Li T,Liu G L,Duan M X,et al. Radish phospholipid hydroperoxide glutathione peroxidase provides protection against hydroperoxide-mediated injury in mouse 3T3 fibroblasts. Bmb REP,2009,42(10):648-654.
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