Cloning and expression analysis of DoUGT83A1 from Dendrobium officinale
Objective To clone one glycosyl transferase gene named DoUGT83A1 from Dendrobium officinale and analyze its bioinformatics information and expression patterns. Methods The DoUGT83A1 gene was obtained from the leaf tissue of D. officinale 'Yandangshan 1'. Online softwares were used for bioinformatics analysis. Phylogenetic tree was constructed using MEGA 11. Gene expression patterns were analyzed by qRT-PCR. Results The DoUGT83A1 gene (GI:1622023669) was cloned by PCR. The CDS sequence of the DoUGT83A1 gene was 1347 bp,encoding 448 amino acids. The DoUGT83A1 protein was a unstable hydrophilic protein,with no signal peptide or transmembrane region. The relative molecular weight of DoUGT83A1 was 54330,and its theoretical isoelectric point (pI) was 5.75. DoUGT83A1 contained 41 phosphorylation sites and two conserved domains (UDPGT,PSPG). Amino acid sequence analysis showed that DoUGT83A1 had the highest homology with UGT83A1 protein of D. nobile,with a similarity of 96.36％. Molecular docking demonstrated the catalytic activity of DoUGT83A1 protein on kaempferol. Tissue-specific expression analysis showed that the relative expression levels of DoUGT83A1 varied across different tissues of 'Yandangshan 1' as follows:leaves>stems>flowers>roots (white roots,green root tips). The results of cis-acting element analysis showed that the promoter sequence of DoUGT83A1 contained multiple elements,which were related to abiotic stress response. And the expression was significantly up-regulated under the induction of low temperature,phosphorus deficiency and mycorrhizal symbiosis,which suggested that this gene may regulate the mycorrhizal symbiosis and abiotic stress response process of D. officinale. Conclusion The cloning,bioinformatics analysis and expression patterns analysis of DoUGT83A1 of D. officinale were completed,which provided a theoretical basis for further exploring the function of DoUGT83A1 and its mechanism in mycorrhizal symbiosis and abiotic stress response.
Dendrobium officinale Kimura et Migo.DoUGT83A1 genebioinformatics analysisexpression characteristic analysisqRT-PCR
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