广西植物2025，Vol.45Issue(9)：1592-1606,15.DOI:10.11931/guihaia.gxzw202408007
秋茄KoNAC25基因的克隆、亚细胞定位及表达分析
Cloning,subcellular localization and expression analysis of KoNAC25 gene in Kandelia obovata
摘要
Abstract
NAC transcription factors are widely present in plants and unique to plants,which play important roles in regulating plant growth and development,hormone signaling,and the response to adverse stress.In our previous work,a low-temperature-induced NAC gene,KoNAC25(GenBank accession number PP860407),was identified from the genome of the mangrove Kandelia obovata.To investigate whether the KoNAC25 gene of K.obovata is involved in the response to salt stress conditions,this study cloned the KoNAC25 gene from the cDNA of K.obovata leaves via RT-PCR and performed sequence characterization,phylogenetic evolution,subcellular localization,and gene expression analyses.The results were as follows:(1)The open reading frame(ORF)of the KoNAC25 gene was 858 bp in length and encoded 285 amino acids,with a molecular weight of 32.9 kDa and a theoretical isoelectric point of 8.53.KoNAC25 was a hydrophilic protein and contained no signal peptide or transmembrane structure,and its secondary structure was predominantly random coils(58.95%).(2)Amino acid sequence homology alignment showed the N-terminus of the KoNAC25 protein had a NAM superfamily structural domain,which consisted of five conserved substructural domains(ABCDE)and belonged to the NAM subfamily.(3)The phylogenetic tree revealed that the KoNAC25 transcription factor of K.obovata was similar to the transcription factors of Ricinus communis,Manihot esculenta,and Hevea brasiliensis.(4)The subcellular localization test confirmed that the protein was localized in the nucleus,which was consistent with bioinformatics prediction.(5)qRT-PCR analysis result revealed that the expression of the KoNAC25 gene was significantly greater in leaves than in flowers and could be induced to upregulate its expression by NaCl(200 mmol·L-1)and abscisic acid(ABA)(100 μmol·L-1),which peaked under 12 h treatment and 6 h treatment,respectively;however,the expression level of this gene was not induced by salicylic acid(100 μmol·L-1)and methyl jasmonate(100 μmol·L-1).In summary,KoNAC25 is involved in the related regulation of the salt stress response in Kandelia obovata,which provides a reference for further research on the function of KoNAC25 gene and its expression regulatory mechanism under salt stress in K.obovata.关键词
秋茄/NAC25基因/基因克隆/亚细胞定位/表达模式Key words
Kandelia obovata/NAC25 gene/gene cloning/subcellular localization/expression pattern分类
农业科技引用本文复制引用
杨党,蒋文骏,王永峰,王建强,董超,杜照奎..秋茄KoNAC25基因的克隆、亚细胞定位及表达分析[J].广西植物,2025,45(9):1592-1606,15.基金项目
浙江省基础公益研究计划项目(GN21C160013) （GN21C160013）
浙江省自然资源厅科技项目(2021-43) （2021-43）
浙江省省级地质专项资金项目([省资]2024010) （[省资]2024010）
台州市科技计划项目(22nya05). （22nya05）
