Abstract
Bacterial canker of kiwifruit caused by Pseudomonas syringae pv. actinidiae was first described in Japan and Korea and is currently an emerging disease that causes major losses in China, Italy, New Zealand, France, Portugal, and Chile. Gold kiwifruit (Actinidia chinensis), especially cvs. Jin Tao and Hort 16A, seem to be more susceptible than green kiwifruit (Actinidia deliciosa) cvs. Hayward and Summer. The bacterium affects male and female woody vines equally, with young vines being more susceptible. The most characteristic symptoms that appear in early spring are reddish orange or white exudates associated with cankers and wounds in branches and/or trunk, as well as brown leaf spots. Buds and fruits were also affected (1). In Spain, 1,132 ha of kiwifruit orchards yielded 25,285 t of fruit in 2009 (2). Most Spanish kiwifruit is cultivated in Galicia (northwest Spain), where the main cultivar is Hayward. In 2010, the first plantation of cv. Jin Tao and one plantation of cv. Summer were established in this area close to Hayward woody vine. In early spring 2011, 80% of the vines in one orchard had twigs with reddish exudates and branches and trunks as well as leaves with angular spots surrounded by yellow haloes. Isolations from both Actinidia spp. were conducted on nutrient agar with sucrose. One hundred and twelve isolates were obtained and seventy-seven were aerobic, gram negative and nonfluorescent on King's B medium. Biochemical tests performed were levan, oxidase, potato rot, arginine didhydrolase, hypersensitivity in tobacco, and utilization of 49 carbohydrates by the API 50 CH system (BioMérieux, Marcy l'Etoile, France). Three PCR protocols were used: two with pathovar-specific primers (PSAF1/PSAR2 and PSAF3/PSAR4) and one with nonspecific primers (PsITSF1/PsITSR2) (3). The results of all biochemical and molecular tests were in agreement with those expected for P. syringae pv. actinidiae. The 16S-23S region of strain EFA 37 isolated from A. deliciosa cv. Summer was sequenced (GenBank Accession No. JF815537) and had 100% sequence identity with P. syringae pv. actinidiae (GenBank Accession Nos. AY342165 and D86357). Pathogenicity tests were performed on 15 plants of A. deliciosa cv. Hayward (five plants per isolate) with the Spanish representative strain EFA 37 and compared with two reference strains isolated from both Actinidia species in Italy and five plants of an untreated control. Three buds per healthy vine were wounded with a sterile needle, inoculated with 30 to 50 μl of each bacterial suspension (108 CFU/ml), sealed, and then covered with plastic. Five leaves per healthy vine were also pierced with a sterile needle and then atomized with the same suspension. Symptoms began to appear after 5 days on inoculated vines, but not on untreated control vines. The bacterium, P. syringae pv. actinidiae, was reisolated from symptomatic plants. The kiwifruit orchard with affected plants was eradicated (25 ha). To our knowledge, this is the first report of P. syringae pv. actinidiae in Spain.
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