First report of powdery mildew caused by Erysiphe corylacearum on hazelnut in Bulgaria.

Abstract

In June 2020, during an observation of fruit-bearing hazelnut trees (Corylus avelana L., cv. Tonda Gentile), grown in a small private home garden in the region of Kavarna (North-Eastern Black Sea coast of Bulgaria), symptoms of powdery mildew were observed on each of the 15 trees. Compared to the widely spread in Bulgaria powdery mildew caused by Phyllactinia guttata and known with its powdery patches and differing chasmothecia exclusively formed on the abaxial leaf surface, the newly observed disease was characterized by the presence of concomitant pathogen's structures on both the abaxial and adaxial surfaces of the leaves, varying from single pustules to 90% severity on certain leaves, as well as on the husks which were 100% affected. Among the white mycelium, cylindrical conidiophores and singly produced conidia (22-34 x 16-22 µm; n = 50) were found. Scattered or in groups, black spherical chasmothecia (diameter of 85-115 µm; n = 30), possessing 8 to 12 appendages with dichotomous branched apices, were also observed in the autumn. Chasmothecia contained 3 to 5 subglobose asci (45-60 x 34-62 µm; n = 30), each ascus containing 8 hyaline and ellipsoid ascospores (15-22 x 9-16 µm; n = 50). From one field powdery mildew sample DNA was extracted using the Nucleospin Plant II kit (Macherey-Nagel, Germany) and the rDNA ITS region was amplified using primers PMITS1 and PMITS2 (Cunnington, J.H. et al., 2003). The amplicon was sequenced (Macrogen Europe, The Netherlands) and used in a BLAST analysis. This resulted in 100% homology with the ITS sequence of Erysiphe corylacearum. The obtained sequence was different from the ones of Erysiphe pseudocorylacearum (99.20% homology) and Erysiphe coryli-americanae (96.52% homology). The targeted sequence was submitted to GenBank (accession number OQ860748). Based on the symptoms, morphological characteristics and molecular identification, the pathogen was identified as Erysiphe corylacearum. Pathogenicity of the fungus was confirmed under lab conditions (natural day/light regime, 23-25°C) by shaking down conidia from naturally infected leaves on both sides of healthy leaves (2-years plantlets, cv. Tonda Gentile, 3 replicates) and incubation in a moist chamber (plastic bags) for the first 24 hours. First small fluffy white pustules appeared 6-7 days after inoculation, subsequently, the disease was progressing and the pathogen's conidiophores and conidia were morphologically identical to that first observed under natural infections in the private garden. The non-inoculated control plantlets (grown under the same conditions in a separate room) remained symptomless during the entire three weeks testing period. To our knowledge, this is the first report of Erysiphe corylacearum on hazelnut in Bulgaria. In October 2021, the same disease was also found in a hazelnut orchard (1.2 ha) located in the region of Pazardzhik (Central Southern part of the country). Globally, this report indicates the disease's expansion in the Black Sea region since its first observation in Turkey in 2013 (Sezer, A., et al. 2017; Heluta, V.P., et al. 2019; Meparishvili, G., et al. 2019; Rosati, M., et al. 2021) and elsewhere in Europe as well (Beenken, L., et al. 2020; Mezzalama, M., et al. 2021; Voglmayr, H., et al. 2021).