The Influence of the Grapevine Bacterial and Fungal Endophytes on Biomass Accumulation and Stilbene Production by the In Vitro Cultivated Cells of Vitis amurensis Rupr.

Abstract

Plant endophytes are known to alter the profile of secondary metabolites in plant hosts. In this study, we identified the main bacterial and fungal representatives of the wild grape Vitis amurensis Rupr. microbiome and investigated a cocultivation effect of the 14 endophytes and the V. amurensis cell suspension on biomass accumulation and stilbene biosynthesis. The cocultivation of the V. amurensis cell culture with the bacteria Agrobacterium sp., Bacillus sp., and Curtobacterium sp. for 2 weeks did not significantly affect the accumulation of cell culture fresh biomass. However, it was significantly inhibited by the bacteria Erwinia sp., Pantoea sp., Pseudomonas sp., and Xanthomonas sp. and fungi Alternaria sp., Biscogniauxia sp., Cladosporium sp., Didymella sp. 2, and Fusarium sp. Cocultivation of the grapevine cell suspension with the fungi Didymella sp. 1 and Trichoderma sp. resulted in cell death. The addition of endophytic bacteria increased the total stilbene content by 2.2–5.3 times, while the addition of endophytic fungi was more effective in inducing stilbene accumulation by 2.6–16.3 times. The highest content of stilbenes in the grapevine cells cocultured with endophytic fungi was 13.63 and 13.76 mg/g of the cell dry weight (DW) after cultivation with Biscogniauxia sp. and Didymella sp. 2, respectively. The highest content of stilbenes in the grapevine cells cocultured with endophytic bacteria was 4.49 mg/g DW after cultivation with Xanthomonas sp. The increase in stilbene production was due to a significant activation of phenylalanine ammonia lyase (PAL) and stilbene synthase (STS) gene expression. We also analyzed the sensitivity of the selected endophytes to eight antibiotics, fluconazole, and trans-resveratrol. The endophytic bacteria were sensitive to gentamicin and kanamycin, while all selected fungal strains were resistant to fluconazole with the exception of Cladosporium sp. All endophytes were tolerant of trans-resveratrol. This study showed that grape endophytes stimulate the production of stilbenes in grape cell suspension, which could further contribute to the generation of a new stimulator of stilbene biosynthesis in grapevine or grape cell cultures.

Effect of VaMyb40 and VaMyb60 Overexpression on Stilbene Biosynthesis in Cell Cultures of Grapevine Vitis amurensis Rupr.

“…Overexpression of the VaMyb40 and VaMyb60 genes did not change the spectrum of detected individual stilbenes ( Table 2 ). As in previously published works [ 39 , 41 ], we detected presence of eight stilbenes. Five stilbenes in all lines were well detectable (more than 0.06 mg/g DW): t -resveratrol diglucoside (1), t -piceid (2), t -resveratrol (3), ε-viniferin (4), δ-viniferin (5), and three stilbenes were usually detected in trace amounts (no more than 0.01 mg/g DW): cis -resveratrol (6), cis -piceid (7), t -piceatannol (8).…”

Fungal Endophytes of Vitis vinifera—Plant Growth Promotion Factors

“…Phoma glomerata, Chaetomium globosum, Aureobasidium pullulans, Epicoccum nigrum, and Acremonium spp. have repeatedly exhibited antibacterial and antifungal properties effective against a number of plant diseases [6,[17][18][19][20][21][22]. Alternaria alternata and Fusarium proliferatum have also been identified as promising biocontrol agents against specific pathological conditions of Vitis vinifera, such as grapevine downy mildew caused by Plasmopara viticola [6,22,23].…”

Resveratrol biosynthesis, optimization, induction, bio-transformation and bio-degradation in mycoendophytes

“…were significantly induced resveratrol production than the rest of the isolated fungi 10.3389/fmicb.2022.1010332 (Yang et al, 2016), suggesting the potential effect of applying endophytic consortium to create different patterns of wine grape chemical profile and the resultant wine. Co-cultivation of tissue culture seedlings with endophytic fungi is similar to their daily growth and it was a potential system to enhance the content of bioactive metabolites (Aleynova et al, 2021). The amount of stilbene and trans-resveratrol was increased in Vitis amurensis cells co-cultured with endophytic fungi Biscogniauxia sp., Cladosporium sp., and Didymella spp.…”

“…Resveratrol is widely distributed in numerous plant types such as grapes ( Vitis vinifera ), berries, peanuts ( Arachis hypogaea ), chocolate, tea, and cassia seeds ( Burns et al, 2002 ; Baur and Sinclair, 2006 ; Wang et al, 2013 ; Mei et al, 2015 ). Grapes are considered the most abundant source of resveratrol content compared to other plants ( Langcake and Pryce, 1977 ; Creasy and Coffee, 1988 ; Aleynova et al, 2021 ). Resveratrol is produced as phytoalexin to protect plants upon exposure to biotic stress such as fungal and bacterial attacks or abiotic stress such as UV radiation ( Adrian et al, 2000 ; Cichewicz et al, 2000 ; Bavaresco et al, 2012 ; Dwibedi and Saxena, 2022 ).…”