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The bioavailability of pollen bioactive compounds for humans is limited. In this study, our aim was to enhance the health-related benefits of pollen by fermentation with a Kombucha/SCOBY (symbiotic culture of bacteria and yeasts) consortium. We performed the fermentation of pollen suspended from the beginning with SCOBY on sweetened green tea or on Kombucha vinegar, by adding pollen after 20 days of Kombucha fermentation. We analyzed: formation of bioactive compounds (anti-oxidant polyphenols, soluble silicon, hydroxy-acids, short chain fatty acids—SCFA); parameters related to Kombucha fermentation (dynamics of lactic acid bacteria—LAB, formation of organic acids, soluble sugar evolution on Kombucha vinegar); the influence of Kombucha fermentation on pollen morphology and ultrastructure; in vitro cytotoxic and antitumoral effects of the Kombucha fermented pollen. The pollen addition increases LAB proportion in the total number of SCOBY microbial strains. SEM images highlight the adhesion of the SCOBY bacteria to pollen. Ultrastructural analysis reveals the release of the pollen content. The content of bioactive compounds (polyphenols, soluble silicon species and SCFA) is higher in the fermented pollen and the product shows a moderate antitumoral effect on Caco-2 cells. The health benefits of pollen are enhanced by fermentation with a Kombucha consortium.

In this work, we developed an enzymatic voltammetric biosensor for the determination of catechin and gallic acid in green tea and kombucha samples. The differential pulse voltammetry (DPV) methodology was optimized regarding the amount of crude enzyme extract, incubation time in the presence of the substrates, optimal pH, reuse of the biosensor, and storage time. Samples of green tea and kombucha were purchased in local markets in the city of Goiânia-GO, Brazil. High performance liquid chromatography (HPLC) and Folin-Ciocalteu spectrophotometric techniques were performed for the comparison of the analytical methods employed. In addition, two calibration curves were made, one for catechin with a linear range from 1 to 60 µM (I = −0.152 * (catechin) − 1.846), with a detection limit of 0.12 µM and a quantification limit of 0.38 µM and one for gallic acid with a linear range from 3 to 60 µM (I = −0.0415 * (gallic acid) − 0.0572), with a detection limit of 0.14 µM and a quantification limit of 0.42 µM. The proposed biosensor was efficient in the determination of phenolic compounds in green tea.

Abstract Kombucha, historically an Asian tea-based fermented drink, has recently become trendy in Western countries. Producers claim it bears health-enhancing properties that may come from the tea or metabolites produced by its microbiome. Despite its long history of production, microbial richness and dynamics have not been fully unraveled, especially at an industrial scale. Moreover, the impact of tea type (green or black) on microbial ecology was not studied. Here, we compared microbial communities from industrial-scale black and green tea fermentations, still traditionally carried out by a microbial biofilm, using culture-dependent and metabarcoding approaches. Dominant bacterial species belonged to Acetobacteraceae and to a lesser extent Lactobacteriaceae, while the main identified yeasts corresponded to Dekkera, Hanseniaspora and Zygosaccharomyces during all fermentations. Species richness decreased over the 8-day fermentation. Among acetic acid bacteria, Gluconacetobacter europaeus, Gluconobacter oxydans, G. saccharivorans and Acetobacter peroxydans emerged as dominant species. The main lactic acid bacteria, Oenococcus oeni, was strongly associated with green tea fermentations. Tea type did not influence yeast community, with Dekkera bruxellensis, D. anomala, Zygosaccharomyces bailii and Hanseniaspora valbyensis as most dominant. This study unraveled a distinctive core microbial community which is essential for fermentation control and could lead to Kombucha quality standardization. Microbial ecology of industrial Kombucha fermentations.

With the development of the tea industry, understanding the differences in root zone microecology among different tea varieties is of great significance for improving tea quality and yield. To investigate the microbial and metabolite foundation underlying the disparities in root zone physicochemical properties between the high-aroma new tea variety JL4 (Jinlong No.4) and its grandparent HD (Huangdan), the bacterial diversity, community structure and metabolite distinctions of HD and JL4 root zone soils were analyzed using NovaSeq 6000 high-throughput sequencing and GC-MS-derived metabolomics technologies. The analysis of soil physical and chemical properties showed that, compared with HD, the available phosphorus (AP) in JL4 was significantly decreased (28.91 ±  9.78 mg · kg −1 , P  < 0.05), and so was the available potassium (AK) at 57.67 ±  4.04 mg · kg −1 . The results from 16S rDNA sequencing indicated that, compared with HD, JL4 had a lower Shannon index and a higher abundance of Gram-negative and aerobic-related bacteria. These results indicated that there was a decrease in bacterial diversity in the root zone soil of JL4. The dominant bacterial phyla included Proteobacteria , Acidobacteriota , and Chloroflexi among others. Biomarkers in HD included Firmicutes , Rhizobiales , and Caulobacterales , and biomarkers for JL4 comprised Sphingomonadaceae bacterium URHD0088 and Halomonadaceae . GC-MS derivatization metabolomics highlighted sugars as having the most differential metabolites (8). In JL4, D-manitol 2 and scylo-inositol decreased while (-)-epicatechin, catechin, and D-pinitol increased. KEGG pathway enrichment analysis revealed substantial enrichment in metabolic pathways related to flavonoid biosynthesis. The changes in these metabolites may have a significant impact on the growth and quality of tea plants. Redundancy Analysis (RDA) along with correlation analyses indicated significant impacts on root zone bacterial community structure by factors such as AK, Soil Organic Matter (SOM), NO 3 − -N (nitrate nitrogen), and pH levels. A significant positive correlation was observed between AK and both Firmicutes and Kapabacteria individually; furthermore, AP exhibited a highly significant positive correlation with Kapabacteria but a significant negative correlation with unidentified Archaea . Catechin and (-)-Epicatechin were significantly negatively correlated with Actinobacteria phylum while showing a significant positive correlation with Verrucomicrobia and Kryptonia phyla. This study systematically compared the microbial and metabolite characteristics of the root zone soil of JL4 and HD for the first time, providing new ideas and methods for tea variety improvement and precision cultivation management, which is expected to promote the high-quality development of the tea industry in the future.

This is the first report of widespread and stress-tolerant PGPR from the tea rhizosphere of the Kangra valley. A total of 493 rhizobacteria were isolated from the major tea-growing regions of the Kangra valley. Molecular fingerprinting of 160 distinct morphotypes using ARDRA and ERIC techniques revealed intergenic and intragenic variability, resulting in the identification of 52 rRNA and 56 ERIC types belonging to 21 distantly related genera, identified by 16S rRNA gene sequencing. Bacillus constituted more than half of the genotypes, followed by Pseudomonas , Burkholderia , Lysinibacillus , Citrobacter , Enterobacter , and Paenibacillus. Bacillus altitudinis , B. cereus , B. megaterium , B. subtilis subsp. inaquosorum , B. methylotropicus , Pseudomonas frederiksbergensis , P. mohnii , and P. moreiii were found to be the most common in the tea rhizosphere across various locations. Quantitative assaying of 42 selected strains revealed significant variations in PGP activities ranging from 55–624 µg/ml for tri-calcium phosphate (TCP) solubilization, 4–3145 nM α-ketobutyrate h/mg/protein ACC-deaminase activity, 2–85 µg/ml IAA-like auxins production, and 2–83% siderophore production. Nine out of 42 PGPR also solubilized aluminium phosphate (Al-P) and iron phosphate (Fe–P). These efficient PGPR are suitable for application in tea soils, which are generally low in available phosphorus, a growth-limiting factor for tea cultivation. Five highly efficient PGPR also showed robust growth under different abiotic stresses under controlled conditions. Inoculum application of 5 efficient and abiotic stress tolerant PGPR showed a significant increment of 1.8–9.4%, 12–16.2%,18.1–30.3% and 21.4–39.2% in plant height, leaf number, fresh and dry weight of tea seedlings under the nursery conditions with 50% reduced NPK concentrations after one year of inoculations, respectively. These selected PGPR genotypes with multifarious PGP activities and natural ability to occur widely can be useful in developing plant microbial inoculants for improving tea productivity.

The aim of the present study is to evaluate plant growth promoting and biocontrol efficacy of a Serratia marcescens strain ETR17 isolated from tea rhizosphere for the effective management of root rot disease in tea. Isolated bacterial culture ETR17 showed significant level of in vitro antagonism against nine different foliar and root pathogens of tea. The phenotypic and molecular characterization of ETR17 revealed the identity of the bacterium as Serratia marcescens. The bacterium was found to produce several hydrolytic enzymes like chitinase, protease, lipase, cellulase and plant growth promoting metabolites like IAA and siderophore. Scanning electron microscopic studies on the interaction zone between pathogen and antagonistic bacterial isolate revealed severe deformities in the fungal mycelia. Spectral analyses (LC-ESI-MS, UV-VIS spectrophotometry and HPLC) and TLC indicated the presence of the antibiotics pyrrolnitrin and prodigiosin in the extracellular bacterial culture extracts. Biofilm formation by ETR17 on polystyrene surface was also observed. In vivo application of talc-based formulations prepared with the isolate ETR17 in tea plantlets under green house conditions revealed effective reduction of root-rot disease as well as plant growth promotion to a considerable extent. Viability studies with the ETR17 talc formulation showed the survivability of the isolate up to six months at room temperature. The sustenance of ETR17 (concentration of 8-9x108 cfu g-1) in the soil after the application of talc formulation was recorded by ELISA. Safety studies revealed that ETR17 did not produce hemolysin as observed in pathogenic Serratia strains. The biocontrol strain reported in this study can be used for field application in order to minimize the use of chemical fungicides for disease control in tea gardens.

Kombucha is a fermented tea made from a Symbiotic Culture of Bacteria and Yeast (SCOBY) with a long history of use as a health tonic. It is likely that most health benefits come from the tea and fermentation metabolites from specific microbial communities. Despite its growing importance as a functional health drink, the microbial ecosystem present in kombucha has not been fully documented. To characterize the microbial composition and biochemical properties of ‘The Good Brew’ original base kombucha, we used metagenomics amplicon (16S rRNA and ITS) sequencing to identify the microbial communities at the taxonomic level. We identified 34 genera with 200 microbial species yet described in kombucha. The dominance of organic acid producing microorganisms Acetobacter, Komagataeibacter and Starmerella are healthy for the human gut and their glucose metabolising activities have a putative role in preventing conditions such as diabetes and obesity. Kombucha contains high protein (3.31 µg/mL), high phenolic content (290.4 mg/100 mL) and low sugars (glucose: 1.87 g/L; sucrose 1.11 g/L; fructose: 0.05 g/L) as compared to green tea. The broad microbial diversity with proven health benefits for the human gut suggests kombucha is a powerful probiotic. These findings are important to improve the commercial value of kombucha and uncover the immense prospects for health benefits.

Fu brick tea is a unique post-fermented tea product which is fermented with microorganism during the manufacturing process. Metabolic analysis showed that most metabolites content were decreased during the manufacturing process of Fu brick tea, except GA (gallic acid). Illumina MiSeq sequencing of ITS gene amplicons was applied to analyze the fungal community succession. The genera Aspergillus , Cyberlindnera and Candida were predominant at the early stage of manufacturing process (from “primary dark tea” to “fermentation for 3 days”), but after the stage of “fermentation for 3 days” only Aspergillus was still dominated, and maintain a relatively constant until to the end of manufacturing process. The effects of metabolites on the structure of the fungal community were analyzed by redundancy analysis (RDA) and variation partitioning analysis (VPA). The results indicated that GCG (gallocatechin gallate), EGCG (epigallocatechin gallate) and GA as well as the interactions among them were the most probably ones to influence, or be influenced by the fungal communities during the fermentation process of Fu brick tea. This study revealed fungal succession, metabolite changes and their relationships, provided new insights into the mechanisms for manufacturing process of Fu brick tea.

Microbial enzymes during solid-state fermentation (SSF), which play important roles in the food, chemical, pharmaceutical and environmental fields, remain relatively unknown. In this work, the microbial communities and enzymes in SSF of Pu-erh tea, a well-known traditional Chinese tea, were investigated by integrated metagenomics/metaproteomics approach. The dominant bacteria and fungi were identified as Proteobacteria (48.42%) and Aspergillus (94.98%), through pyrosequencing-based analyses of the bacterial 16S and fungal 18S rRNA genes, respectively. In total, 335 proteins with at least two unique peptides were identified and classified into 28 Biological Processes and 35 Molecular Function categories using a metaproteomics analysis. The integration of metagenomics and metaproteomics data demonstrated that Aspergillus was dominant fungus and major host of identified proteins (50.45%). Enzymes involved in the degradation of the plant cell wall were identified and associated with the soft-rotting of tea leaves. Peroxiredoxins, catalase and peroxidases were associated with the oxidation of catechins. In conclusion, this work greatly advances our understanding of the SSF of Pu-erh tea and provides a powerful tool for studying SSF mechanisms, especially in relation to the microbial communities present.

This study aimed at isolation of native plant growth-promoting bacteria (PGPB) associated with organic tea plantations. Most research on tea and associated microbes have been on Darjeeling and Assam, known for their world-class tea. However, emerging tea plantations in remote Northeast India are gaining prominence due to their unique geographical location, favorable climate, and organic practices. This study investigated PGBP associated with these organic tea plantations, aimed to assess their potential cross-infectivity on non-host plants. A total of 58 PGP bacterial isolates were isolated from four organic tea plantations. Six potential isolates were further evaluated individually and as consortium for their PGP on rice and maize. Bacillus, Pseudomonas, and Serratia spp. as individual and in consortium were found to have potent cross-infectivity with significant growth promotion in non-host plants indicated by plant height, root length, shoot, and root weight. The present findings suggest that PGPB native to organic tea plantations have potential cross-infectivity for use as a biofertilizers to improve the growth and productivity of non-host crops. This provides prospectives of using native bacteria on non-host plants paving the way for their potential application in sustainable agriculture practices for growth promotion of staple food crops.