PJB-2024-222
Molecular characterization and expression analysis of dovin2 gene in dendrobium officinale cold and drought stress response
Boting Liu
Abstract
Sucrose metabolism is a crucial metabolic process in plants that involves acid invertase. Nonetheless, the precise function and structural characteristics of the vacuolar invertase gene in Dendrobium officinale remain unclear. In this study, the putative vacuolar invertase gene (DoVIN2) was isolated from the DanXia cultivar using homologous cloning techniques. DoVIN2 exhibited a cDNA sequence spanning 1368 bp, encoding a protein consisting of 455 amino acids with a glycosyl hydrolase domain. Sequence analyses revealed DoVIN2 as a stable, hydrophilic protein lacking a signal peptide or transmembrane region, but possessing 46 phosphorylation sites. Phylogenetic analysis underscored the conservation of plant VINVs during evolution, with DoVIN2 clustering with this evolutionary lineage. Furthermore, GFP expression assays in rice protoplasts localized DoVIN2 to the cell nucleus. Tissues-specific expression analysis revealed varying levels of DoVIN2 expression across tissues, with flower exhibiting the highest expression followed by stem, root, and leaf. The DoVIN2 promoter region contained abiotic stress–responsive elements. Under cold stress, DoVIN2 expression increased, accompanied by soluble sugar content, indicating its involvement in polysaccharide accumulation and cold stress response. Conversely, DoVIN2 expression exhibited a negative correlation with the duration of drought stress, implying its diverse roles in responding to cold and drought stress through distinct signaling pathways. Additionally, in silico protein-protein interaction network data revealed that DoVIN2 interacts with enzymes implicated in saccharometabolism. These findings suggest that DoVIN2 plays a crucial role in abiotic stress response by regulating the expression of enzyme genes, thereby enhancing D. officinale's tolerance to adverse conditions. Overall, this study provides valuable insights into the function of DoVIN2 and lays a foundation for investigation into D. officinale's response to abiotic stress.
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