Sustainable under-vine dry mulching with cover crop residues enhances above- and below-ground microbial biodiversity in a rainfed vineyard (Vitis vinifera L.)

Abstract

In Mediterranean rainfed vineyards, cover cropping and dry mulching are increasingly implemented to improve soil structure, microclimate, and vine resilience, yet their effects on grapevine-associated microbiomes remain only partially understood. This study evaluated the influence of two winter cover crop mixtures on soil and plant microbial communities: a cereal-based mixture (CM) and a balanced mixture (BM) including cereals, legumes, and brassicas. A traditional system with alternate tillage and spontaneous grass cover served as the control (C). Cover crops were terminated at full flowering in spring, and their residues were retained as under-vine dry mulch.

Root and leaf samples collected at grapevine pre-flowering and veraison were used to characterize rhizosphere and phyllosphere microbial communities through high-throughput sequencing targeting bacterial (16S rRNA) and fungal (ITS) regions. At veraison, dry mulching improved soil microclimate by increasing moisture and reducing temperature by about 2.5 °C. CM particularly enhanced vine physiological performance. These conditions supported higher rhizosphere richness and enriched nutrient-cycling taxa such as Bradyrhizobium sp. and Nitrospira japonica, whereas C favored drought-tolerant PGPR including Bacillus zanthoxyli, Gaiella occulta, Roseiflexus sp., Pseudarthrobacter sp., and Paenibacillus sp. In the phyllosphere, both CM and BM reduced Erysiphe necator abundance and increased Aureobasidium pullulans, a potential biocontrol agent. Overall, cover crop-derived dry mulching enhanced soil microclimate, microbial diversity, and vineyard resilience under rainfed conditions.

References

Abad, J., Hermoso De Mendoza, I., Marín, D., Orcaray, L., & Santesteban, L. G. (2021). Cover crops in viticulture. A systematic review (1): Implications on soil characteristics and biodiversity in vineyard. OENO One, 55(1), 295–312. https://doi.org/10.20870/OENO-ONE.2021.55.1.3599

Ambrosini, A., & Passaglia, L. M. P. (2017). Plant growth–promoting bacteria (PGPB) in the rhizosphere. In B. Lugtenberg (Ed.), Principles of Plant–Microbe Interactions (Vol. 2, pp. 190–209). Wiley.

Bozoudi, D., & Tsaltas, D. (2018). The multiple and versatile roles of Aureobasidium pullulans in the vitivinicultural sector. Fermentation, 4(4), 85. https://doi.org/10.3390/fermentation4040085

Burg, P., Čížková, A., Mašán, V., Sedlar, A., Matwijczuk, A., & Souček, J. (2022). The effect of mulch materials on selected soil properties, yield and grape quality in vineyards under Central European conditions. Agronomy, 12(8), 1862.

Castellano-Hinojosa, A., & Strauss, S. L. (2020). Impact of cover crops on the soil microbiome of tree crops. Microorganisms, 8(3), 328. https://doi.org/10.3390/microorganisms8030328

Celette, F., Gaudin, R., & Gary, C. (2008). Spatial and temporal changes to the water regime of a Mediterranean vineyard due to the adoption of cover cropping. European Journal of Agronomy, 29(4), 153–162. https://doi.org/10.1016/j.eja.2008.04.007

Cunial, L., Diti, I., Bonini, P., Patelli, R., Gatti, M., Cola, G., & Poni, S. (2025). Under-vine mulching with winter cover crop residues improves water status, ecophysiology and berry quality of cv. Barbera grapevines. European Journal of Agronomy, 164, 127514. https://doi.org/10.1016/j.eja.2025.127514

Lorenz, D. H., Eichhorn, K. W., Bleiholder, H., Klose, R., Meier, U., & Weber, E. (1995). Growth stages of the grapevine (Vitis viniferaL. ssp. vinifera): Codes and descriptions according to the extended BBCH scale. Australian Journal of Grape and Wine Research, 1(2), 100–103.

Lu, Y., Zhou, G., Ewald, J., Pang, Z., Shiri, T., & Xia, J. (2023). MicrobiomeAnalyst 2.0: Comprehensive statistical, functional and integrative analysis of microbiome data. Nucleic Acids Research, 51(W1), W310–W318. https://doi.org/10.1093/nar/gkad288

Novara, A., Catania, V., Tolone, M., Gristina, L., Laudicina, V. A., & Quatrini, P. (2020). Cover crop impact on soil organic carbon, nitrogen dynamics and microbial diversity in a Mediterranean semiarid vineyard. Sustainability, 12(8), 3256.

Perazzolli, M., Antonielli, L., Storari, M., et al. (2014). Resilience of the natural phyllosphere microbiota of the grapevine to chemical and biological pesticides. Applied and Environmental Microbiology, 80(12), 3585–3596. https://doi.org/10.1128/AEM.00415-14

Steenwerth, K., & Belina, K. M. (2008). Cover crops enhance soil organic matter, carbon dynamics and microbiological function in a California vineyard. Applied Soil Ecology, 40(2), 359–369.

Vorholt, J. A. (2012). Microbial life in the phyllosphere. Nature Reviews Microbiology, 10(12), 828–840.

Vukicevich, E., Lowery, T., Bowen, P., Úrbez-Torres, J. R., & Hart, M. (2016). Cover crops to increase soil microbial diversity and mitigate decline in perennial agriculture: A review. Agronomy for Sustainable Development, 36, 48.

Zarraonaindia, I., Owens, S. M., Weisenhorn, P., et al. (2015). The soil microbiome influences grapevine-associated microbiota. mBio, 6(2), e02527-14. https://doi.org/10.1128/mBio.02527-14

Acknowledgments

This study was partially supported by ‘National Research Centre for Agricultural Technologies (Agritech)’ CN00000022.PNRR-M4C2, Investment 1.4 -Funded by the European Union -NextGenerationEU’ -CUP J33C22001160007. This manuscript reflects only the authors’ views and opinions, neither the European Union nor the European Commission can be considered responsible for them. The study were supported by the Doctoral School on the Agro-Food System (Agrisystem) of the faculty of agriculture of Università Cattolica del Sacro Cuore, Piacenza, Italy.