Association of Dryocola boscaweniae, Gibbsiella greigii and Gibbsiella quercinecans with oak decline in Iran

Tree decline is described as the loss in tree vigor and increased mortality initiated by climate change events, and also involves pathogens and pests. Stem bleeding and bark canker of oak (Quercus castaneifolia) were observed in Mazandaran and Golestan forests during summer 2020–2021. Symptoms included cracks in the outer bark, stem tissue necrosis, bleeding, dark exudate and dark brown lesions in the inner bark. Eighty-nine strains with a metallic green sheen pigment were isolated on eosin methylene blue agar. The pathogenicity of all strains recovered in this study was assessed on oak seedlings and acorns. Forty-four strains produced rotting on oak acorns 2 weeks after inoculation. Inoculation of six representative strains on oak seedlings resulted in twig dieback of the plants after 4 weeks. Strains were negative for Gram reaction, oxidase and levan formation from sucrose. The gyrB and infB gene sequence similarity values of strains were 98.87–99.57% with the type strain of Gibbsiella quercinecans, 98.66–98.86% with the type strain of Gibbsiella greigii and 99.46–99.64% with the type strain of Dryocola boscaweniae. In the phylogenetic tree based on concatenated sequences of gyrB and infB genes or each gene individually, the strains were divided into three clusters containing the type strains of G. quercinecans, G. greigii and D. boscaweniae, each with high bootstrap support and confirming their identity as belonging to these three species. To the best of our knowledge, this is the first report of oak bacterial canker caused by D. boscaweniae, G. greigii and G. quercinecans in Iran and the first report of D. boscaweniae associated with oak decline symptoms.