A new bacterial disease of Persian (English) walnut (Juglans regia) has been observed in France. This disease, called vertical oozing canker (VOC), is characterized by vertical cankers on trunks and branches of affected walnut trees with oozing exudates. To determine the aetiology of the disease, a study was carried out in 79 walnut orchards and nurseries located in southeastern and southwestern France. Bacterial analysis from diseased samples yielded 36 strains identified as Xanthomonas arboricola and 32 strains identified as Brenneria nigrifluens on the basis of biochemical tests. The causal agent of VOC was identified as X. arboricola by pathogenicity tests on walnut. Fluorescent amplified fragment length polymorphism (F-AFLP) was carried out on 36 strains of X. arboricola collected in this study, 24 strains of X. arboricola pv. juglandis isolated from walnut blight symptoms and one strain of X. arboricola pv. corylina included as an outgroup. Based on cluster analysis of F-AFLP data, most X. arboricola strains responsible for main VOC outbreaks showed a high degree of similarity, forming a cluster clearly separate from strains of X. arboricola pv. juglandis isolated from walnut blight symptoms. It is suggested that VOC is caused by a distinct genetic lineage within the pathovar juglandis of X. arboricola that is also able to cause classical bacterial blight symptoms on walnut leaves and fruits.

Since the summer of 2000, vertical oozing cankers have been observed on trunks and branches of Persian walnut trees (Juglans regia). Cvs. Fernor, Chandler, Mayette, and Hartley were the most frequently affected, but cvs. Lara and Franquette could also be affected. Brenneria nigrifluens (synonym Erwinia nigrifluens) (3) was isolated from diseased trees from 13 orchards and nurseries in southwestern (Aquitaine, Périgord, Charentes, and Quercy), southeastern (Grenoble areas), and western (near Angers) France. Cankers were observed on trunks and branches where brown-to-black exudates staining the bark appeared mainly in the summer. Isolations were performed primarily from exudates but also from infected tissues by using King's medium B. Colonies similar in appearance to Brenneria nigrifluens (1) were purified and characterized. Gram reaction, Kovac's oxidase, oxidative-fermentative metabolism, and urease activity were assayed for all isolates. API Biotype 100 kits (BioMérieux, Marcy l'Etoile, France) were used as recommended, except that incubations were at 28°C for 4 days. When compared with the reference strain (French Collection of Plant Pathogenic Bacteria (CFBP) 4998T = National Collection of Plant Pathogenic Bacteria (NCPPB) 564T = American Type Culture Collection (ATCC) 13028T) from California, 14 isolated strains were identified as B. nigrifluens on the basis of physiological and biochemical characteristics. These 14 strains were deposited in the CFBP under Accession Nos. 6746 to 6759. Pathogenicity of three selected strains (CFBP 6746, 6747, and 6758) was confirmed by inoculating branches of 7-year-old walnut trees with 108 CFU of each isolate introduced in wounds (2). The reference strain (CFBP 4998T) and water were similarly inoculated as controls. Two and five months later, necrotic lesions were observed in the inner bark and dark lines were observed in internal wood, but no external cankers were observed on any trees inoculated with the local and reference strains. B. nigrifluens was reisolated from the dark lines in internal wood up to approximately 10 cm from the inoculation site. To our knowledge, this is the first report of this bacterium in France. References: (1) L. Hauben et al. Syst. Appl. Microbiol. 21:384, 1998. (2) M. Ridé and S. Ridé. Proc. Int. Conf. Plant Pathogenic Bacteria, 4th, 2:957, 1978. (3) E. E. Wilson et al. Phytopathology 47:669, 1957.

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