Managing the phyllosphere: Agronomic practices influence the ecology and evolution of Pseudomonas syringae in cherry orchards

Zeng, Z.; Mansfield, J. W.; Vadillo-Dieguez, A.; Connell, J.; Irvine, J.; Hulin, M. T.; Stavridou, E.; Karlstrom, A.; Frutos, F. D.; Grinberg, N. F.; Rabiey, M.; Harrison, R. J.; Xu, X.; Jackson, R. W.

doi:10.64898/2026.04.22.719873

bioRxiv · 10.64898/2026.04.22.719873

Managing the phyllosphere: Agronomic practices influence the ecology and evolution of Pseudomonas syringae in cherry orchards

Zeng, Z.·Mansfield, J. W.·Vadillo-Dieguez, A.·Connell, J.·Irvine, J.·Hulin, M. T.·Stavridou, E.·Karlstrom, A.·Frutos, F. D.·Grinberg, N. F.·Rabiey, M.·Harrison, R. J.·Xu, X.·Jackson, R. W.

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Abstract

Bacterial canker, caused the Pseudomonas syringae species complex, is a major constraint on sweet cherry production worldwide. However, the influence of agronomic practices on pathogen ecology, dispersal and evolution under field conditions remains poorly understood. Here, we combined a factorial-design field experiment with whole-genome sequencing to investigate the effects of polytunnel covering and nitrogen fertigation on phyllosphere populations and the dynamics of a key pathogen, P. syringae pathovar syringae 9644 (Pss9644) in young cherry trees. Epiphytic P. syringae populations initially resembled those in surrounding woodland environments. Over time, pathogenic phylogroup 2d lineages became dominant, particularly on uncovered trees. Diversity of P. syringae populations was higher in uncovered treatments. Polytunnel covering markedly altered community composition and limited rain-splash dispersal of Pss9644 from stem cankers to leaves, thereby interrupting a key stage of the disease cycle. By contrast, nitrogen fertigation had no detectable effect on phyllosphere community structure, but enhanced plant growth and reduced lesion expansion following inoculation. Whole-genome sequencing of re-isolated Pss9644 strains revealed limited short-term genomic diversification, with single-nucleotide polymorphisms detected in 22 re-isolates. In total, 36 mutations were identified across the chromosome although no mutation affected virulence or motility. Taken together, our results show that agronomic practices influence both pathogen ecology and disease outcomes through distinct mechanisms: polytunnel covering primarily limits pathogen dispersal and reshapes phyllosphere communities, while nitrogen fertigation enhances plant growth and reduces disease severity. These findings highlight the potential to integrate canopy management and nutrient strategies to mitigate bacterial canker risk in commercial cherry production.

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Zeng, Z., Mansfield, J. W., Vadillo-Dieguez, A., Connell, J., Irvine, J., Hulin, M. T., Stavridou, E., Karlstrom, A., Frutos, F. D., Grinberg, N. F., Rabiey, M., Harrison, R. J., Xu, X., Jackson, R. W.. 2026-04-22. Managing the phyllosphere: Agronomic practices influence the ecology and evolution of Pseudomonas syringae in cherry orchards. https://doi.org/10.64898/2026.04.22.719873

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