First Report ofTruncatella angustataCausing Postharvest Rot on ‘Topaz’ Apples in the Netherlands

Abstract

In the Netherlands, about 30% of the organic apple (Malus domestica Borkh.) production consists of apple scab resistant cultivars, such as Topaz and Santana. However, organic ‘Topaz’ apples show a high incidence of fungal rot after storage. Hot-water treatment (HWT) of freshly harvested apple fruit prior to long-term storage is an important strategy for the control of postharvest diseases, especially in the organic production sector (Maxin et al. 2012). The recommended treatment temperatures and times vary according to the cultivar because of the risk of heat damage to the fruit peel. In January 2016, light peel damage caused by HWT was observed on ‘Topaz’ apples from an organic orchard. Also, up to 15% of the ‘Topaz’ apples showed typical rot lesions of an unknown causal agent. The lesions showed brown, irregular necrosis and were slightly sunken. To isolate the causal agent, fruits were rinsed with sterile water, lesions were sprayed with 70% ethanol until droplet runoff, the skin was removed aseptically with a scalpel, and tissue under the lesion was placed onto potato dextrose agar (PDA). The PDA plates were incubated at 20°C in the dark, and single spore isolates were transferred to fresh PDA plates. The colonies that appeared on PDA were cottony to woolly, dull white to brown in color, with black acervuli mainly in the center of the PDA plates. The isolates produced four-celled conidia, 16 to 19 × 7 to 9 µm, straight to slightly curved, with two brown to dark-brown median cells that had thick walls. More than one hyaline apical appendage, variable in size and branched dichotomically, were observed and a basal appendage was absent. The fungus was morphologically identical to Truncatella angustata (Pers.) S. Hughes (Sutton 1980). The identity of two representative isolates (PPO-45246 and PPO-45321) was confirmed by means of gene sequencing. To this end, DNA was extracted using the LGC Mag Plant Kit (Berlin, Germany) in combination with the Kingfisher method (Waltham, MA). Sequences of the ITS region were amplified using primers ITS1/ITS4, sequenced, and deposited in GenBank under accession numbers KX085227 and KX085228. MegaBLAST analysis revealed that both of our ITS sequences matched 99% with T. angustata isolates in GenBank (EU342216, JX390614, and KF646105). Koch’s postulates were fulfilled using 10 ‘Topaz’ apples. Surface sterilized fruits were inoculated with 20 μl of 10 5 conidiospores ml –1 in water, prepared from a 15-day-old PDA culture of the isolate PPO-45246, after wounding with a needle. Inoculated fruits were sealed in a plastic bag and incubated in darkness at 20°C. Symptoms appeared after 7 days on 100% of the fruits while mock-inoculated controls with water remained symptomless. Fungal colonies isolated from the lesions and cultured on PDA morphologically resembled the inoculated isolates. The identity of the reisolations was confirmed as T. angustata by sequencing. T. angustata has a worldwide distribution and has also been reported to cause leaf spot on Rosa canina (Eken et al. 2009), canker and twig dieback on blueberry (Vaccinium spp.) (Espinoza et al. 2008), and fruit rot of olive (Olea europaea) (Arzanlou et al. 2012). To the best of our knowledge, this is the first report of T. angustata causing fruit rot of apples. Importantly, we note that the occurrence of this fruit rot may be enhanced by wounding, in this case as a result of hot water treatment.