First report of Neopestalotiopsis rosae causing black root rot on strawberry (Fragaria × ananassa) in Albania

In December 2022, 30% strawberry (Fragaria × ananassa) plants of Savana, Sabrina, and Fortuna cultivars grown in greenhouses showed symptoms of leaf necrosis, root rot, and wilt. Small spots with light brown center and dark edges appeared on the leaves, then enlarging and coalescing into irregular necrotic lesions. Disease symptoms appeared also on ripe fruits with light to dark brown spots. Pure cultures of fungal colonies were obtained from the margin of infected leaf tissues of ten diseased plants collected from Kafaraj, Fier (40°37′28.7″N 19°31′03.5″E). Black acervuli (180 μm diameter) appeared scattered on the surface of 7–10 old-days colonies. Conidia were spindleshaped, five-celled, four-septate, 8.0 μm width and 26.0 μm length, with hyaline apical and basal cells and brown middle cells, unbranched appendages arising from apex (32 μm) and a small filament at the basal cell (10 μm). All colonies were morphologically identified as belonging to Neopestalotiopsis genus (Maharachchikumbura et al. 2014). Four strains, PEST-1, PEST-2, PEST-3 and PEST-4, stored in the microbial ITEM Collection of CNR-ISPA, were molecularly identified by sequencing the informative markers internal transcribed spacer, β-tubulin and elongation factor, with primer pairs and PCR conditions reported in Norphanphoun et al. (2019). FASTA sequences showed 100% homology with N. rosae CBS 101057, by BLAST search. Phylogenetic analysis, with Maximum Likelihood method, proved that all strains belong to N. rosae. Sequences were deposited in the GenBank database (accession numbers PP348710- PP348717 and PP339777-PP339780). Pathogenicity was tested on healthy strawberry seedlings of Fortuna cultivar, inoculating the roots with 106 spore/ml suspensions, in triplicate. The strains used for pathogenicity test were reisolated from symptomatic plants, fulfilling Koch’s postulates. Several studies demonstrated that Neopestalotiopsis species can cause root and crown rots in strawberry, representing an emerging problem worldwide (EPPO 2024). To our knowledge, this is the first report of Neopestalotiopsis rosea causing black root rot of strawberry in Albania.