Neofusicoccum parvum and Diaporthe foeniculina associated with twig and shoot blight and branch canker of citrus in Greece.
In a survey performed in Chania and Aetoloacarnania, Greece in years 2013-2014, fungal isolates causing twig and shoot blight and branch canker of citrus trees were morphologically characterized and identified by multiple gene sequence analysis. By sequencing the ITS-5.8S rRNA, the elongation factor 1-α (EF1-α), the β-tubulin and the RNA polymerase II subunit (Rpb2) genes, the isolates examined were associated with Diaporthe foeniculina (six isolates) and Neofusicoccum parvum (one isolate). All six D. foeniculina isolates showed slow colony growth rates (7.4±3.2 mm/day), while the N. parvum isolate exhibited fast growth (41.6 mm/day). Koch's criteria were met after re-isolation of D. foeniculina isolates from all inoculated Citrus spp. and N. parvum from inoculated C. reticulata "Ortanique" and after having developed symptoms similar to those detected on shoots and branches collected from citrus fields. Based on lesion length on detached C. medica "Lia Kritis" shoots, N. parvum caused long necrotic lesions (58 mm in length) in comparison with a length of 12-21 mm lesions caused by D. foeniculina isolates. Pathogenicity trials on nine Citrus spp., which had been inoculated with D. foeniculina and N. parvum, revealed different levels of susceptibility, indicating a host-dependent infection effect, with Poncirus trifoliate × C. paradisi ("Citrumelo Swingle") being the most resistant citrus genotype. Lack of host specificity suggests that their pathogen-host association could be attributed to ecological rather to co-evolutionary factors. This work represents the first report, accompanied with pathogenicity tests, on botryosphaeriaceous and diaporthaceous pathogens associated with twig and shoot blight and branch canker of citrus in Greece.