谷子转录因子基因SibZIP42在拟南芥中对高盐和ABA的响应

doi:10.3864/j.issn.0578-1752.2016.17.003

Abstract

Abstract: 【Objective】 This study was conducted to analyze the molecular characteristics and biological function of the transcription factor SibZIP42 in foxtail millet and to discuss the regulation pathway of SibZIP42 in improving salt tolerance. At the same time, the study would provide a potential gene for improving abiotic stress resistance in crop molecular breeding. 【Method】 Bioinformatics methods were used to analyze the molecular characteristics of SibZIP42. DNAMAN and MEGA 5.05 softwares were used to do multiple sequences alignment and construct the phylogenetic tree of SibZIP42. The 2 000 bp sequence upstream of SibZIP42 was downloaded as the promoter sequence from databases Phytozome, and the cis-acting elements of SibZIP42 promoter were analyzed in PLACE database. NetPhos 2.0 Server database was used to predict phosphorylation sites of SibZIP42 protein. The real-time PCR was used to analyze the expression patterns of SibZIP42 under various stress treatments. SibZIP42 was fused with GFP to detect its subcellular localization in protoplast cells. SibZIP42 was overexpressed in Arabidopsis to analyze its function under high salt and ABA stress conditions. 【Result】 The full length of SibZIP42 was 546 bp with one exons and encode a hydrophilic protein with 181 amino acid residues, and the protein molecular weight was about 20.3 kD. Phylogenetic tree showed that the gene is located in the S subgroup of bZIP family. It is the highest sequence homology with AtbZIP42. Promoter cis-acting element analysis showed that there are many stress-related response elements including ABRE, MYB and MYC in promoter sequence of SibZIP42. Phosphorylation site analysis showed that there are 14 serine, 4 tyrosine and 1 threonine phosphorylation sites in SibZIP42 protein sequence. The expression pattern analysis showed that SibZIP42 is involved in responses to various abiotic stresses and exogenous hormones. The gene expression profile results indicated that the expression level of SibZIP42 in root was higher than stem and leaf, and it was induced by drought, salt and ABA treatments in millet. The protein subcellular localization analysis revealed that SibZIP42 protein was localized in nucleus. Functional analysis of SibZIP42 showed that there is no significant difference of germination rate between transgenic lines and wild-type Arabidopsis WT on the normal MS medium, whereas on the MS culture medium with NaCl concentration of 90, 120 and 150 mmol·L^-1, germination rate of SibZIP42 transgenic Arabidopsis was significantly higher than WT. Under 90 mmol·L^-1 NaCl treatment condition, cotyledon green rate of SibZIP42 transgenic Arabidopsis was significantly higher than WT, whereas cotyledon green rate of SibZIP42 transgenic Arabidopsis was significantly lower than WT when the concentration of ABA was 0.5, 1 and 2 μmol·L^-1 in MS culture medium. Some ABA-related response genes including HIS1-3, RD29B and RAB18 and drought-related response gene including PIP2A were up-regulated in SibZIP42 transgenic Arabidopsis, which suggested that the overexpression of SibZIP42 improved salt stress tolerance in transgenic Arabidopsis by ABA signaling pathway. 【Conclusion】 The salt tolerance of SibZIP42 transgenic Arabidopsis was significantly stronger than WT during the period of seed germination, while SibZIP42 transgenic Arabidopsis seeds were more sensitive to ABA treatment than WT which indicated that SibZIP42 positively regulated salt tolerance in transgenic Arabidopsis by ABA signaling pathway.

Key words: foxtail millet (Setaria italica L.), bZIP transcription factor, salt stress tolerance, ABA signaling pathway

QIN Yu-hai, ZHANG Xiao-hong, FENG Lu, LI Wei-wei, XU Zhao-shi, LI Lian-cheng, ZHOU Yong-bin, MA You-zhi, DIAO Xian-min, JIA Guan-qing, CHEN Ming, MIN Dong-hong. Response of Millet Transcription Factor Gene SibZIP42 to High Salt and ABA Treatment in Transgenic Arabidopsis[J].Scientia Agricultura Sinica, 2016, 49(17): 3276-3286.

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Response of Millet Transcription Factor Gene SibZIP42 to High Salt and ABA Treatment in Transgenic Arabidopsis

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