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“Golden flower” fungi in dark tea are beneficial to human health. The rapid identification method of “golden flower” fungi can verify the quality of dark tea products and ensure food safety. In this study, 6 strains were isolated from Liupao tea. They were respectively identified as A. cristatus, A. chevalieri, and A. pseudoglaucus. A. pseudoglaucus was reported as Liupao tea “golden flower” fungus for the first time. It was found that the ITS and BenA sequences of A. cristatus and A. chevalieri were highly conserved. It is difficult to clearly distinguish these closely related species by ITS sequencing. To rapidly identify species, multiplex PCR species-specific primers were designed based on orphan genes screened by comparative genomics analysis. Multiplex PCR results showed that orphan genes were specific and effective for the identification of A. cristatus and A. chevalieri isolated from Liupao tea and Fu brick tea. We confirmed that orphan genes can be used for identification of closely related Aspergillus species. Validation showed that the method is convenient, rapid, robust, sequencing-free, and economical. This promising method will be greatly beneficial to the dark tea processing industry and consumers.

Liupao tea, a popular dark tea, exhibited anti-diabetic activity in Chinese tea culture. However, the bioavailability of the main active components is low. Therefore, the relationship between the anti-diabetic activity and the modulation of gut microbiota was investigated in this study for the first time. Liupao tea extract (LTE) could significantly alleviate hyperglycemia, insulin-resistance, and plasma lipid profiles. The hypoglycemic effect of high-dose LTE was comparable to that of metformin. Importantly, LTE exhibited beneficial effect on modulating the diabetic-induced disorder of gut microbiota, such as increasing Bacteroidetes/Firmicutes ratio and up-regulating putative short-chain fatty acid (SCFA)-producing bacteria, including Prevotella and Bacteroides. Moreover, 29 key phylotypes were significantly correlated with at least one diabetic index. In conclusion, LTE could be developed into an anti-diabetic functional beverage and its beneficial effects is likely to be associated with the prebiotic effects for increasing the level of SCFAs and the relative abundance of SCFA-producing bacteria.

From humanistic characteristics, historical origin, fame, quality, and the correlation between quality and production region, we analyze key factors of protection for Geographic Indication product Liupao Tea. We also analyze industrial development of Liupao Tea from processing threshold, scale of processing enterprises, standard and index, planting base, research and development and technology popularization, and pesticide control. In line with these situations, we put forward following suggestions. First, it is proposed to raise standard index and formulate national standard for LiuDao Tea. Second, we should popularize the industrial mode of “market + association enterprise) + base” and establish demonstration area for protection of Geographic Indication product Liupao Tea. Third, it is recommended to raise the production and processing threshold. Fourth, we should establish special production inspection and research and development institution. Fifth, it is suggested to set up independent supervision organization.