Biomics

Research of the environmentally safe biological methods of stimulating the growth of medicinal and edible mushrooms assists the development of scientific foundations of culture technologies. This work poses a task of characterizing the effect of bacteria from the Azospirillum genus, which are known by their phyto-stimulating properties, upon the growth parameters of xylotrophic basidiomycete Flammulina velutipes under the conditions of dual fungal-bacterial cultures. The possibility of submerged co-cultivation of the winter mushroom F. velutipes with the endophytic and epiphytic strains of Azospirillum brasilense was demonstrated, and optimal conditions for the co-cultivation were selected. Mycelial development in joint cultures was explored in dependence of media composition; inoculum characteristics, and bacterial strain. Under the conditions of liquid co-cultures optimized by both the medium composition and the bacterial inoculum concentration, the azospirilla strains under study had active mobility and formed clusters near fungal hyphae. The concentration of bacterial cells had a significant effect on the growth of binary cultures. Co-cultivation of F. velutipes with А. brasilense Sp245 at the optimal concentration (0.5%, v/v) of bacterial cells in the inoculum (А₆₀₀ = 1) allowed to obtain the dry-biomass value of 2.35 times higher relative to the control. The intensive growth of a mixed culture of mushrooms with azospirillum under study was promoted by the medium based on Glc and Fru (in mass proportion 1 : 1), and Asn. The data gained are indicative of the great potentialities of binary mushroom-bacterial cultures application for obtaining efficiently the mycelial biomass of basidiomycete.

mushrooms, basidiomycetes, Flammulina, PGPR, Azospirillum, co-culture, submerged culture

1. Alves M.I., Macagnan K.L., Rodrigues A.A., de Assis D.A., Torres M.M., de Oliveira P., Furlan L., Vendruscolo C.T., Moreira A.D.S. Poly (3-hydroxybutyrate)-P (3HB): Review of production process technology. Industrial Biotechnology. V. 13(4). P. 192–208. doi:10.1089/ind.2017.0013
2. Arkhipova T.N., Kuzmina L.Y., Kudoyarova G.R. The influence of the titer of microorganisms on the results of their growth-stimulating activity in the heterotrophic nutrition of plants. Ecobiotech. 2018. V. 1(2). P. 97–104. doi 10.31163/2618-964X-2018-1-2-97-104
3. Benaissa A. Plant Growth Promoting Rhizobacteria A review. Algerian Journal of Environmental Science and Technology. V. 5(1). P. 873–880.
4. Berthold D.E., Shetty K.G., Jayachandran K., Laughinghouse IV H.D., Gantar M. Enhancing algal biomass and lipid production through bacterial co-culture. Biomass and Bioenergy. 2019. V. P. 280–289. doi 10.1016/j.biombioe.2019.01.033
5. Fukami J., Cerezini P., Hungria M. Azospirillum: benefits that go far beyond biological nitrogen fixation. AMB Express. 2018. 8(1). P. 73. doi:10.1186/s13568-018-0608-1
6. Goswami M., Deka S. Plant growth-promoting rhizobacteria – alleviators of abiotic stresses in soil: a review. Pedosphere. V. 30(1). P. 40–61. doi:10.1016/S1002-0160(19)60839-8
7. Huang L.H., Lin H.Y., Lyu Y.T., Gung C.L., Huang C.T. Development of a transgenic Flammulina velutipes oral vaccine for hepatitis B. Food Technology and Biotechnology. 2019. V. 57(1). P. 105–112. doi:10.17113/ftb.57.01.19.5865
8. Ivanchina N.V., Garipova S.R., Shavaleeva D.V., Urazbakhtina N.A., Zakharova R.Sh., Khairullin R.M. Effect of Bacillus subtilis Strains on the Yield of Pea (Pisum sativum) at Monoinoculation and in Combination with Rhizobium leguminosarum bv. viceae 1078. Agrokhimiya. 2008. (10). 34–39.
9. Janusz G., Pawlik A., Sulej J., Świderska-Burek U., Jarosz-Wilkołazka A., Paszczyński A. Lignin degradation: microorganisms, enzymes involved, genomes analysis and evolution. FEMS Microbiology Reviews. 2017. V. 41(6). P. 941–962. doi:10.1093/femsre/fux049
10. Jong S.C., Donovick R. Antitumor and antiviral substances from fungi. Advances in Applied Microbiology. 1989. 34. P. 183–262. doi:10.1016/S0065-2164(08)70319-8
11. Kumar A., Patel J.S., Meena V.S., Ramteke P.W. Plant growth-promoting rhizobacteria: strategies to improve abiotic stresses under sustainable agriculture. Journal of Plant Nutrition. 2019. 42(11-12). P. 1402–1415. doi:10.1080/01904167.2019.1616757
12. Kupryashina M.A., Petrov S.V., Ponomareva E.G., Nikitina V.E. Ligninolytic activity of bacteria of the genera Azospirillum and Niveispirillum. Microbiology. 2015. V. 84(6). P. 791–795. doi:10.1134/S0026261715060041
13. Lasa A.V., Mašínová T., Baldrian P., Fernández-López M. Bacteria from the endosphere and rhizosphere of Quercus use mainly cell wall-associated enzymes to decompose organic matter. PloS ONE. 2019. V. 14(3). P. e0214422. doi 10.1371/journal.pone.0214422
14. Loiko N.G., Kryazhevskikh N.A., Suzina N.E., Demkina E.V., Muratova A.Y., Turkovskaya O.V., Kozlova A.N., Galchenko V.F., El'-Registan G.I. Resting forms of Sinorhizobium meliloti. Microbiology. 2011. V. 80(4). P. 472–482. doı:10.1134/S0026261711040126
15. Loshchinina E.A., Tsivileva O.M., Makarov O.E., Nikitina V.E. Changes in carbohydrate and fatty-acid content of Lentinus edodes mycelium in dual cultures with Azospirillum brasilense. Izvestiya vuzov. Prikladnaya khimiya i biotekhnologiya. 2012. (2-3). P. 64–67.
16. Mahfuz S., Song H., Miao Y., Liu Z. Dietary inclusion of mushroom (Flammulina velutipes) stem waste on growth performance and immune responses in growing layer hens. Journal of the Science of Food and Agriculture. V. 99(2). P. 703–710. doi:10.1002/jsfa.9236
17. Mizuno T. Bioactive biomolecules of mushrooms: food function and medicinal effect of mushroom fungi. Food Reviews International. 1995. V. 11(1). P. 5-21. doi:10.1080/87559129509541017
18. Naureen Z., Rehman N.U., Hussain H., Hussain J., Gilani S.A., Al Housni S.K., Mabood F., Khan A.L., Farooq S., Abbas G., Harrasi A.A. Exploring the potentials of Lysinibacillus sphaericus ZA9 for plant growth promotion and biocontrol activities against phytopathogenic fungi. Frontiers in Microbiology. 2017. V. P. 1477. doi:10.3389/fmicb.2017.01477
19. Nie Y., Jin Y., Deng C., Xu L., Yu M., Yang W., Li B., Zhao R. Rheological and microstructural properties of wheat dough supplemented with Flammulina velutipes (mushroom) powder and soluble polysaccharides. CyTA-Journal of Food. V. 17(1). P. 455–462. doi:10.1080/19476337.2019.1596986
20. Nikitina V.E., Tsivileva O.M., Loshchinina E.A. Interrelations of xylotrophic basidiomycetes and soil nitrogen-fixing bacteria from the genus Azospirillum. Advances in Medical Mycology. 2006. V. 7. P. 293–294.
21. Paaventhan P., Joseph J.S., Seow S.V., Vaday S., Robinson H., Chua K.Y., Kolatkar P.R. A 1.7 Å structure of Fve, a member of the new fungal immunomodulatory protein family. Journal of Molecular Biology. V. 332(2). P. 461–470. doi:10.1016/S0022-2836(03)00923-9
22. Palacios O.A., Lopez B.R., Bashan Y., de-Bashan L.E. Early Changes in Nutritional Conditions Affect Formation of Synthetic Mutualism Between Chlorella sorokiniana and the Bacterium Azospirillum brasilense. Microbial Ecology. 2019. V. 77(4). P. 980– doi:10.1007/s00248-018-1282-1
23. Seow S.V., Kuo I.C., Paaventhan P., Kolatkar P.R., Chua K.Y. Crystallization and preliminary X-ray crystallographic studies on the fungal immunomodulatory protein Fve from the golden needle mushroom (Flammulina velutipes). Acta Crystallographica Section D: Biological Crystallography. V. 59(8). P. 1487–1489. doi:10.1107/S0907444903011879
24. Su A., Ma G., Xie M., Ji Y., Li X., Zhao L., Hu Q. Characteristic of polysaccharides from Flammulina velutipes in vitro digestion under salivary, simulated gastric and small intestinal conditions and fermentation by human gut microbiota. International Journal of Food Science & Technology. 2019. 54(6). P. 2277–2287. doi:10.1111/ijfs.14142
25. Tsivileva O.M., Pankratov A.N., Nikitina V.E. Extracellular protein production and morphogenesis of Lentinula edodes in submerged culture. Mycological Progress. 2010. V. 9(2). P. 157–167. doi:10.1007/s11557-009-0614-4
26. Tsivileva O.M., Shaternikov A.N., Nikitina E. Bacteria of the Azospirillum genus for the optimization of the artificial culture of xylotrophic mushrooms. Biotekhnologiya. 2020. V. 36(2). P. 16–25. doi:10.21519/0234-2758-2020-36-2-16-25
27. Wasser S.P. Medicinal mushrooms as a source of antitumor and immunomodulating polysaccharides. Applied Microbiology and Biotechnology. V. 60(3). P. 258–274. doi:10.1007/s00253-002-1076-7
28. Wasser S.P., Weis A.L. Medicinal properties of substances occurring in higher basidiomycetes mushrooms: current perspectives. International Journal of Medicinal Mushrooms. 1999. 1(1). P. 31–62. doi:10.1615/IntJMedMushrooms.v1.i1.30
29. Wasser S.P., Weis A.L. Therapeutic effects of substances occurring in higher Basidiomycetes mushrooms: a modern perspective. Critical Reviews in Immunology. 1999a. 19(1). P. 65–96. doi:10.1615/CritRevImmunol.v19.i1.30
30. Zhao R., Hu Q., Ma G., Su A., Xie M., Li X., Chen G., Zhao L. Effects of Flammulina velutipes polysaccharide on immune response and intestinal microbiota in mice. Journal of Functional Foods. V. 56. P. 255–264. doi:10.1016/j.jff.2019.03.031