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Functional analysis of a novel C-glycosyltransferase in the orchid Dendrobium catenatum.

ABSTRACT: Flavonoids, which are a diverse class of phytonutrients, are used by organisms to respond to nearly all abiotic stresses and are beneficial for human health. Glycosyltransferase, used during the last step of flavonoid biosynthesis, is important in flavonoid enrichment. However, little is known about glycosyltransferase in the orchid Dendrobium catenatum (D. officinale). In this study, we isolated a novel C-glycosyltransferase (designated DcaCGT) from the orchid D. catenatum by identifying and analyzing 82 putative genes in the GT1 family. DcaCGT could specifically catalyze not only di-C-glycosylation but also O-glycosylation. Apart from the normal function of catalyzing 2-hydroxynaringenin and phloretin to the respective di-C-glycosides, DcaCGT also catalyzes apigenin to cosmosiin. Targeted metabolic profiling of the substrates (2-hydroxynaringenin, phloretin, and apigenin) and products (vitexin, isovitexin, vicenin-2, nothofagin, 3',5'-di-C-glucosylphloretin, and cosmosiin) in different tissues showed that vicenin-2 was the most abundant product of this novel enzyme. Cosmosiin was detected in flowers and flower buds. We also established that DcaCGT functions expanded throughout the evolution of D. catenatum. Residual OGT activity may help D. catenatum resist drought stress. Our study illustrates the function, origin, and differentiation of DcaCGT and provides insights into glycosylation and molecular propagation processes, which can be used to improve the production of flavonoids by the cultivated medicinal plant D. catenatum.

SUBMITTER: Ren Z

PROVIDER: S-EPMC7326982 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:ObjectivesDendrobium catenatum Lindl. (DH) is a Chinese herbal medicine, which is often used to make tea to improve immunity in China. Rumor has it that DH has a protective effect against cardiovascular disease. However, it is not clear how DH can prevent cardiovascular disease, such as atherosclerosis (AS). Therefore, the purpose of this study is to study whether DH can prevent AS and the underlying mechanisms.MethodsZebrafish larvae were fed with high-cholesterol diet (HCD) to establish a zebrafish AS model. Then, we used DH water extracts (DHWE) to pretreat AS zebrafish. The plaque formation was detected by HE, EVG, and oil red O staining. Neutrophil and macrophage counts were calculated to evaluate the inflammation level. Reactive oxygen species (ROS) activity, malondialdehyde (MDA) content, and superoxide dismutase (SOD) activity in zebrafish were measured to reflect oxidative stress. The cholesterol accumulation and the levels of lipid, triglyceride (TG), and total cholesterol (TC) were measured to reflect lipid metabolism disorder. Then, parallel flow chamber was utilized to establish a low shear stress- (LSS-) induced endothelial cell (EC) dysfunction model. EA.hy926 cells were exposed to LSS (3 dyn/cm2) for 30 min and treated with DHWE. The levels of ROS, SOD, MDA, glutathione (GSH), and glutathiol (GSSG) in EA.hy926 cells were analysed to determine oxidative stress. The release of nitric oxide (NO), endothelin-1 (ET-1), and epoprostenol (PGI2) in EA.hy926 cells was measured to reflect EC dysfunction. The mRNA expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) in EA.hy926 cells was detected to reflect EC dysfunction inflammation.ResultsThe results showed that DHWE significantly reduced cholesterol accumulation and macrophage infiltration in early AS. Finally, DHWE significantly alleviate the lipid metabolism disorder, oxidative stress, and inflammation to reduce the plaque formation of AS zebrafish larval model. Meanwhile, we also found that DHWE significantly improved LSS-induced EC dysfunction and oxidative stress in vitro.ConclusionOur results indicate that DHWE could be used as a prevention method to prevent AS.

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Functional analysis of a novel C-glycosyltransferase in the orchid Dendrobium catenatum.

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