广西植物2025，Vol.45Issue(3)：542-554,13.DOI:10.11931/guihaia.gxzw202305051
文采报春苣苔PwDREB2s基因的克隆与表达分析
Cloning and expression analysis of PwDREB2s genes in Primulina wentsaii
摘要
Abstract
Dehydration responsive element binding protein 2(DREB2)is a transcription factor,which plays a regulatory role to drought,high salinity and high temperature responding in model plants.However,it is still unclear for the function of DREB2 genes in Primulina wentsaii,which is adapted to limestone karst habitats with drought-high temperature environment.DREB2 homologous genes responding to dual stresses of drought and high temperature,were screened in P.wentsaii.It could provide a new gene pool for the creation of resistant germplasm in Gesneriaceae.In this study,specific primers were designed based on transcriptome sequences of P.wentsaii firstly,gDNA and cDNA were used as templates to isolate PwDREB2 genes.Then,amino acid sequences,phylogenetic relationships and consensus motifs were analyzed by bioinformatics methods.As well,the expression patterns of PwDREB2s genes were analyzed by semi-quantitative reverse transcription PCR(sqRT-PCR)with single stress,such as low temperature,heat shock,simulated drought(osmosis),high salinity,exogenous ABA and oxidation,respectively.Finally,the transcript levels of designated members,which could respond to single stresses and drought-heat shock combined stresses,were detected with different drought-heat shock patterns by quantitative real-time PCR(qRT-PCR).The results were as follows:(1)Eight intron-lacking PwDREB2s genes were isolated,they could encode 198-386 amino acids,which contained several typical characteristic regions such as AP2/ERF domain,nuclear localization signal or transcriptional activation domain,respectively;(2)Transcription factors,such as PwDREB2A/2AL1/2AL2,PwDREB2D/2DL and PwDREB2F were classified into subtype 1,subtype 2 and subtype 3 of A-2 subgroup,respectively,while PwDREB2EL1/2EL2 were classified into A-6 subgroup;(3)Semi-quantitative RT-PCR analysis showed that the transcript levels of PwDREB2s genes were induced by a series of single stresses,among which PwDREB2A/2AL1/2AL2/2D genes could simultaneously respond to simulated drought and heat shock;(4)qRT-PCR results revealed that the PwDREB2D gene,which was responding to drought-heat shock combined stresses,was tissue-specific,it could only be strongly induced in rhizomes.Compared with mock group,the transcriptional responses of PwDREB2A/2AL1/2AL2 genes were significantly enhanced under natural drought-heat shock combined stresses with soil matrix,except for PwDREB2D gene.However,when it was compared with the single stress of natural drought and heat shock,the expression levels of PwDREB2AL1/2AL2 genes were significantly higher in natural drought-heat shock combined stresses.In conclusion,PwDREB2AL1/2AL2 genes may play the important regulatory role in response to dual stresses of natural drought and high temperature in P.wentsaii.关键词
文采报春苣苔/DREB2转录因子/肉质叶/干旱-热激复合胁迫/表达模式Key words
Primulina wentsaii/DREB2 transcription factor/fleshy leaf/drought-heat shock combined stresses/expression pattern分类
农业科技引用本文复制引用
刘宝骏,付传明,苏江,冼康华,何金祥,黄宁珍..文采报春苣苔PwDREB2s基因的克隆与表达分析[J].广西植物,2025,45(3):542-554,13.基金项目
中央引导地方科技发展资金项目(桂科ZY21195035) （桂科ZY21195035）
广西科学院基本业务费项目(CQZ-E-1910) （CQZ-E-1910）
广西喀斯特植物保育与恢复生态学重点实验室课题(20-065-7). （20-065-7）
