Modulation of Aroma and Sensory Properties of Prokupac Wines by a Bacillus-based Preparation Applied to Grapes Prior to Harvest

M. Malićanin, B. Danilović, D. Cvetković, S. Stamenković-Stojanović, N. Nikolić, M. Lazić, I. Karabegović


Modern viticulture requires the replacement of hazardous agrochemicals with eco-friendly, bio-based products such as microbial preparations that  enhance grape and wine quality while protecting the grapevine from pest and disease attacks. This study investigated the effects of a commercially available Bacillusbased preparation on the volatile and sensory properties of wines made from Vitis vinifera, cv. Prokupac grapes. Three different concentrations of preparation based on Bacillus subtilis Ch-13 were applied to grapevines two weeks prior to harvest. The total soluble solids in the grapes was affected by the application of B. subtilis Ch-1 and the alcohol content of the wine made from these grapes was greater. Wines made
from the B. subtilis Ch-13-treated grapes showed an average increase in total phenolic compounds of about 27%, compared to the wine made from the untreated control grapes. The colour intensity of wines from the
treated grapes, independently of the concentration, was higher by more than 30% than for the wine from the control grape sample. The B. subtilis Ch-13 treatment also affected the content of 3-methyl-1-butanol,
ethyl decanoate and ethyl octanoate in the wine, at about 35%, 40% and 20%, respectively. The latter compounds are responsible for floral and fruity aromas. Generally, wines made from the treated grapes showed similar sensorial characteristics but scored better overall than the control. Principal component analysis showed a clear differentiation between wine made from the control and that from the B. subtilis Ch-13-treated grapes. The results suggest that the application of B. subtilis Ch-13 to grapevines two weeks prior to harvest has a positive effect on wine quality.


Bacillus subtilis; HS-SPME-GC-MS; aroma profile; sensory characteristics; Prokupac

Full Text:



Abbey, J.A., Percival, D., Abbey, L., Asiedu, S.K., Prithiviraj, B. & Schilder, A. 2019. Biofungicides as alternative to synthetic fungicide control of grey mould (Botrytis cinerea)–prospects and challenges. Biocontrol Sci. Technol. 29(3), 241–262.

Aleixandre-Tudo, J.L., Buica, A., Nieuwoudt, H., Aleixandre, J.L. & Du Toit, W. 2017. Spectrophotometric analysis of phenolic compounds in grapes and wines. J. Agric. Food Chem. 65(20), 4009-4026.

Alori, E.T. & Babalola, O.O. 2018. Microbial inoculants for improving crop quality and human health in Africa. Front. Microbiol. 9, 2213.

Aoki, T., Aoki, Y., Ishiai, S., Otoguro, M. & Suzuki, S. 2017. Impact of Bacillus cereus NRKT on grape ripe rot disease through resveratrol synthesis in berry skin. Pest Manag. Sci. 73(1), 174–180.

Aznar, M., López, R., Cacho, J. & Ferreira, V. 2003. Prediction of aged red wine aroma properties from aroma chemical composition. Partial least squares regression models. J. Agric. Food Chem. 51(9), 2700–2707.

Babincev, L.M., Gurešić, D.M. & Simonović, R.M. 2016. Spectrophotometric characterization of red wine color from the vineyard region of Metohia. J. Agric. Sci. 61(3), 281–290.

Biarnès, A., Bailly, J.S. & Boissieux, Y. 2009. Identifying indicators of the spatial variation of agricultural practices by a tree partitioning method: The case of weed control practices in a vine growing catchment. Agric. Syst. 99(2–3), 105–116.

Bleve, G., Tufariello, M., Vetrano, C., Mita, G. & Grieco, F. 2016. Simultaneous alcoholic and malolactic fermentations by Saccharomyces cerevisiae and Oenococcus oeni cells co-immobilized in alginate beads. Front. Microbiol. 7, 943.

Bogicevic, M., Maras, V., Mugoša, M., Kodžulović, V., Raičević, J., Šućur, S. & Failla, O. 2015. The effects of early leaf removal and cluster thinning treatments on berry growth and grape composition in cultivars Vranac and Cabernet Sauvignon. Chem. Biol. Technol. Agric. 2(1), 13.

Calvo-Garrido, C., Roudet, J., Aveline, N., Davidou, L., Dupin, S. & Fermaud, M. 2019. Microbial antagonism toward botrytis bunch rot of grapes in multiple field tests using one bacillus ginsengihumi strain and formulated biological control products. Front. Plant Sci. 10, 15.

Castilhos, M.B.M. de Cattelan, M.G., Conti-Silva, A.C. & Del Bianchi, V.L. 2013. Influence of two different vinification procedures on the physicochemical and sensory properties of Brazilian non-Vitis vinifera red wines. LWT - Food Sci. Technol. 54(2), 360–366.

Chambers IV, E. & Koppel, K. 2013. Associations of volatile compounds with sensory aroma and flavor: The complex nature of flavor. Molecules. 18(5), 4887-905.

Chebotar, V.K., Makarova, N.M., Shaposhnikov, A.I., & Kravchenko, L. V. 2009. Antifungal and phytostimulating characteristics of Bacillus subtilis Ch-13 rhizospheric strain. Prikl. Biokhim. Mikrobiol. 45(4), 465-469.

Coradini, R., Madoşă, E., Irina, P. & Cristina, C. 2014. Change of hue and intensity of color during the fermentation in case of must obtained from various varieties of red grapes from Minis-Maderat Winery. Journal of Horticulture, Forestry and Biotechnology. 18(4), 55–62.

Cosme, F., Gonçalves, B., Bacelar, E.A., Ines, A., Jordão, A.M. & Vilela, A. 2017. Genotype, environment and management practices on red/ dark-colored fruits phenolic composition and its impact on sensory attributes and potential health benefits. In: Soto-Hernández, M. (ed) Phenolic Compounds - Natural Sources, Importance and Applications. InTech. London, UK.

Damalas, C.A. & Koutroubas, S.D. 2018. Current status and recent developments in biopesticide use. Agric. 8(1), 13.

De-la-Fuente-Blanco, A., Sáenz-Navajas, M.P. & Ferreira, V. 2016. On the effects of higher alcohols on red wine aroma. Food Chem. 210, 107–114.

Elmer, P.A.G. & Reglinski, T. 2006. Biosuppression of Botrytis cinerea in grapes. Plant Pathol. 55(2), 155–177.

Escribano-Viana, R., López-Alfaro, I., López, R., Santamaría, P., Gutiérrez, A.R. & González-Arenzana, L. 2018. Impact of chemical and biological fungicides applied to grapevine on grape biofilm, must, and wine microbial diversity. Front. Microbiol. 9, 59.

Esteban, M.., Villanueva, M.. & Lissarrague, J.. 2002. Relationships between different berry components in Tempranillo (Vitis vinifera L) grapes from irrigated and non-irrigated vines during ripening. J. Sci. Food Agric. 82(10), 1136–1146.

González-Rodríguez, J., Pérez-Juan, P. & Luque De Castro, M.D. 2002. Method for the simultaneous determination of total polyphenol and anthocyan indexes in red wines using a flow injection approach. Talanta. 56, 53–59.

Guerrero, R.F., Cantos-Villar, E., Ruiz-Moreno, M.J., Puertas, B., Cuevas, F.J. & Moreno-Rojas, J.M. 2019. Influence of vertical training systems on warm climate red winemaking: Wine parameters, polyphenols, volatile composition, and sensory analysis. Oeno One. 53(3), 471–486.

Heydari, A. & Pessarakli, M. 2010. A review on biological control of fungal plant pathogens using microbial antagonists. J. Biol. Sci. 10(4), 273-290.

Jackson, R.S. 2008. Wine Science: Principles and applications. Elsevier. London: UK.

Jordão, A., Vilela, A. & Cosme, F. 2015. From sugar of grape to alcohol of wine: sensorial impact of alcohol in wine. Beverages. 1(4), 292–310.

Kim, B.S. & Hwang, B.K. 2007. Microbial fungicides in the control of plant diseases. J. Phytopathol. 155(11–12), 641–653.

Lakićević, S., Popović, T., Matijašević, S., Ćirković, B., Lazić, M. & Popović-Đorđević, J. 2019. Chemical evaluation of autochthonous variety “Prokupac” red wine with the addition of selected aromatic herbs. Ann. Univ. Craiova - Agric. Mont. Cadastre Ser. 49(1), 87–97.

Lakićević, S.H., Popović Djordjević, J.B., Pejin, B., Djordjević, A.S., Matijašević, S.M. & Lazić, M.L. 2018. An insight into chemical composition and bioactivity of “Prokupac” red wine. Nat. Prod. Res. DOI: 10.1080/14786419.2018.1516219.

Lambrechts, M.G. & Pretorius, I.S. 2000. Yeast and its importance to wine aroma - A review. S. Afr. J. Enol. Vitic. 21(1), 97–129.

Malićanin, M., Rac, V. & Rakić, V. 2017. The effect of inactivated yeast-based products on the process of wine aging, phenolic compounds and sensory characteristics of red wine Prokupac. BIO Web Conf. 9, 02004.

Marković, N., Pržić, Z., Rakonjac, V., Todić, S., Ranković-Vasić, Z., Matijašević, S. & Bešlić, Z. 2017. Ampelographic characterization of Vitis cv “ Prokupac ” clones by multivariate analysis. Rom. Biotechnol. Lett. 22(5),12868–12875.

Mckay, M. & Buica, A. 2020. Factors influencing olfactory perception of selected off-flavour- causing compounds in red wine - A Review. S. Afr. J. Enol. Vitic. 41(1), 56–71.

Mozell, M.R. & Thachn, L. 2014. The impact of climate change on the global wine industry: Challenges & solutions. Wine Econ. Policy. 3(2014), 81-89.

Nedelkovski, D., Cvetković, J., Beleski, K. & Poposka, H. 2017. Phenolic composition of Vranec grapevine cultivar (Vitis vinifera L.) grafted on different rootstock. Bulg. J. Agric. Sci. 23(3), 389–395.

Niculescu, V.C., Paun, N. & Ionete, R.E. 2018. The evolution of polyphenols from grapes to wines. In: Jordão A.M. (ed) Grapes and wines - Advances in production, processing, analysis and valorization. InTech. London, UK.

OIV. 2019. Compendium of international methods of wine and must analysis. 18, Rue D'aguesseau – 75008 Paris.

Otoguro, M. & Suzuki, S. 2018. Status and future of disease protection and grape berry quality alteration by micro-organisms in viticulture. Lett. Appl. Microbiol. 67(2), 106–112.

Pagliarini, E., Laureati, M. & Gaeta, D. 2013. Sensory descriptors, hedonic perception and consumer’s attitudes to Sangiovese red wine deriving from organically and conventionally grown grapes. Front. Psychol. 4, 896.

Pertot, I., Caffi, T., Rossi, V., Mugnai, L., Hoffmann, C., Grando, M.S., Gary, C., Lafond, D., Duso, C., Thiery, D., Mazzoni, V. & Anfora, G. 2017. A critical review of plant protection tools for reducing pesticide use on grapevine and new perspectives for the implementation of IPM in viticulture. Crop Prot. 97, 70–84.

Pineau, B., Barbe, J.C., Leeuwen, C. Van & Dubourdieu, D. 2009. Examples of perceptive interactions involved in specific “Red-” and “Black-berry” aromas in red wines. J. Agric. Food Chem. 57(9), 3702–3708.

Raicevic, V., Sivcev, B., Jakovljevic, M., Antic, S. & Lalevic, B. 2004. The influence of the biofertilizer type on wine quality and soil microbiological activity. Proceedings of the 1st international symposium on grapevine growing, commerce and research, Lisbon, Portugal.

Rolli, E., Marasco, R., Saderi, S., Corretto, E., Mapelli, F., Cherif, A., Borin, S., Valenti, L., Sorlini, C. & Daffonchio, D. 2017. Root-associated bacteria promote grapevine growth: from the laboratory to the field. Plant Soil. 410(1–2), 369–382.

Sawant, I.S., Wadkar, P.N., Rajguru, Y.R., Mhaske, N.H., Salunkhe, V.P., Sawant, S.D. & Upadhyay, A. 2016. Biocontrol potential of two novel grapevine associated Bacillus strains for management of anthracnose disease caused by Colletotrichum gloeosporioides. Biocontrol Sci. Technol. 26(7), 964–979.

Shafi, J., Tian, H. & Ji, M. 2017. Bacillus species as versatile weapons for plant pathogens: a review. Biotechnol. Biotechnol. Equip. 31(3), 446–459.

Shi, P., Song, C., Chen, H., Duan, B., Zhang, Z. & Meng, J. 2018. Foliar applications of iron promote flavonoids accumulation in grape berry of Vitis vinifera cv. Merlot grown in the iron deficiency soil. Food Chem. 253, 164–170.

Sivčev, B., Jović, S., Raičević, V., Petrović, A. & Lalević, B. 2005. Application of microbiological fertilizers in viticulture: Grape yield and quality of wine cv. Riesling. J. Agric. Sci. 50(1), 19–26.

Stamenković, S., Beškoski, V., Karabegović, I., Lazić, M. & Nikolić, N. 2018. Microbial fertilizers: A comprehensive review of current findings and future perspectives. Spanish J. Agric. Res. 16 (1), e09R01.

Tangolar, S., Alkan Torun, A., Tarım, G., Ada, M., Aydın, O., Kaçmaz, S. & Tangolar, S. 2019. The Effect of Microbial Fertilizer Applications on Grape Yield, Quality and Mineral Nutrition of Some Early Table Grape Varieties. Selcuk J. Agric. Food Sci. 33(2), 62–66.

Thiollet-Scholtus, M., Caillé, S., Samson, A., Lambert, J.J. & Morlat, R. 2014. Use of production practices and sensory attributes to characterize Loire Valley red wines. Am. J. Enol. Vitic. 65(1), 50–58.

Vilanova, M., Genisheva, Z., Masa, A. & Oliveira, J.M. 2010. Correlation between volatile composition and sensory properties in Spanish Albariño wines. Microchem. J. 95(2), 240–246.

Vilela, A., Schuller, D., Mendes-Faia, A. & Côrte-Real, M. 2013. Reduction of volatile acidity of acidic wines by immobilized Saccharomyces cerevisiae cells. Appl. Microbiol. Biotechnol. 97(11), 4991–5000.

Welke, J.E., Zanus, M., Lazzarotto, M. & Alcaraz Zini, C. 2014. Quantitative analysis of headspace volatile compounds using comprehensive two-dimensional gas chromatography and their contribution to the aroma of Chardonnay wine. Food Res. Int. 59, 85–99.

Yadav, S.S., Redden, R.J., Hatfield, J.L., Lotze-Campen, H. & Hall, A.J.W. 2011. Crop adaptation to climate change. John Wiley & Sons, Inc.

Yildirim, E., Karlidag, H., Turan, M., Dursun, A. & Goktepe, F. 2011. Promotion of Broccoli by Plant Growth Promoting Rhizobacteria. Hort Sci. 46(6), 932–936.

Yilmaztekin, M. 2014. Characterization of potent aroma compounds of cape gooseberry (Physalis peruviana L.) fruits grown in antalya through the determination of odor activity values. Int. J. Food Prop. 17(3), 469–480.

Zheng, Y., Tian, L., Liu, H., Pan, Q., Zhan, J. & Huang, W. 2009. Sugars induce anthocyanin accumulation and flavanone 3-hydroxylase expression in grape berries. Plant Growth Regul. 58(3), 251–260.

Zhu, F., Du, B. & Li, J. 2016. Aroma Compounds in Wine. In: Morata, A. (ed.) Grape and Wine Biotechnology, InTech. London, UK.

Zoecklein, B.W., Fugelsang, K.C., Gump, B.H. & Nury, F.S. 1995. Wine analysis and production. Springer, Boston, MA



  • There are currently no refbacks.