中国农业科学2016，Vol.49Issue(17)：3275-3285,11.DOI:10.3864/j.issn.0578-1752.2016.17.003
谷子转录因子基因SibZIP42在拟南芥中对高盐和ABA的响应
Response of Millet Transcription Factor GeneSibZIP42 to High Salt and ABA Treatment in Transgenic Arabidopsis
摘要
Abstract
Objective] This study was conducted to analyze the molecular characteristics and biological function of the transcription factorSibZIP42 in foxtail millet and to discuss the regulation pathway ofSibZIP42in 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 ofSibZIP42. DNAMAN and MEGA 5.05 softwares were used to do multiple sequences alignment and construct the phylogenetic tree ofSibZIP42. The 2 000 bp sequence upstream of SibZIP42 was downloaded as the promoter sequence from databases Phytozome, and the cis-acting elements ofSibZIP42 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 ofSibZIP42 under various stress treatments. SibZIP42 was fused with GFP to detect its subcellular localization in protoplast cells.SibZIP42 was overexpressed inArabidopsisto analyze its function under high salt and ABA stress conditions.[Result] The full length ofSibZIP42 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 withAtbZIP42. Promoter cis-acting element analysis showed that there are many stress-related response elements including ABRE, MYB and MYC in promoter sequence ofSibZIP42. 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 thatSibZIP42 is involved in responses to various abiotic stresses and exogenous hormones. The gene expression profile results indicated that the expression level ofSibZIP42 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 ofSibZIP42 showed that there is no significant difference of germination rate between transgenic lines and wild-typeArabidopsis 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 ofSibZIP42 transgenicArabidopsis was significantly higher than WT. Under 90 mmol·L-1 NaCl treatment condition, cotyledon green rate ofSibZIP42transgenicArabidopsis was significantly higher than WT, whereas cotyledon green rate ofSibZIP42 transgenicArabidopsiswas significantly lower than WT when the concentration of ABA was 0.5, 1 and 2μmol·L-1 in MS culture medium. Some ABA-related response genes including HIS1-3,RD29B andRAB18 and drought-related response gene includingPIP2Awere up-regulated inSibZIP42 transgenic Arabidopsis, which suggested that the overexpression ofSibZIP42 improved salt stress tolerance in transgenicArabidopsis by ABA signaling pathway.[Conclusion] The salt tolerance ofSibZIP42 transgenicArabidopsiswas significantly stronger than WT during the period of seed germination, whileSibZIP42 transgenicArabidopsis seeds were more sensitive to ABA treatment than WT which indicated thatSibZIP42 positively regulated salt tolerance in transgenicArabidopsisby ABA signaling pathway.关键词
谷子/bZIP类转录因子/耐盐胁迫/ABA信号途径Key words
foxtail millet (Setaria italica L.)/bZIP transcription factor/salt stress tolerance/ABA signaling pathway引用本文复制引用
秦玉海,贾冠清,陈明,闵东红,张小红,冯露,李微微,徐兆师,李连城,周永斌,马有志,刁现民..谷子转录因子基因SibZIP42在拟南芥中对高盐和ABA的响应[J].中国农业科学,2016,49(17):3275-3285,11.基金项目
国家转基因生物新品种培育重大专项（2014ZX08002-003B）、陕西省科技统筹创新工程计划（2014KTZB02-01-01）、中国农业科学院创新工程 （）
