Diagnosis and detection of the main bacterial pathogens of stone fruit and almond

Diagnosis and detection are key aspects related to plant health status. A critical review of the available diagnostic methods utilised for Agrobacterium rhizogenes and A. tumefaciens, Pseudomonas amygdali, P. yringae pv. mors-prunorum, P. syringae pv. persicae, P. syringae pv. syringae and Xanthomonas arboricola pv. pruni, the main pathogens of the stone fruit trees, is presented. As there is a general lack of updated standardized protocols for the detection of most of these bacteria, the most appropriate media for their isolation are reported along with serological reagents, PCR and real-time PCR protocols with comments on their accuracy for the analysis of these pathogens in plant samples. There are many selective media for isolation, especially for Agrobacterium spp., but fewer for Pseudomonas spp. and X. arboricola pv. pruni. Serological techniques are not very useful for these pathogens due to the current lack of specific antibodies commercially available. As to molecular methods, it is surprising to find so many PCR protocols for Agrobacterium species, very few and unspecific for the Pseudomonas species pathogenic to stone fruit trees, and several recent PCR protocols for X. arboricola pv. pruni. The new advances in genomics and proteomics will provide information for selecting new targets to develop specific and sensitive techniques for the diagnosis and detection of these bacterial pathogens in plant material.The authors thank the contributions of the participants in the EU-COST Action 873 and the project RF2009-00002-C04-01 “Prospección, ecolección, conservación y caracterización de nuevo germoplasma de melocotonero”

Detection and identification methods and new tests as developed and used in the framework of cost 873 for bacteria pathogenic to stone fruits and nuts

Xanthomonas arboricola pv. pruni (Xap), the causal agent of bacterial spot disease of stone fruits and almond, is regulated as a quarantine pathogen in the European Union and the European and Mediterranean Plant Protection Organization (EPPO). Xap can have an epiphytic phase and/or be latent and, consequently, it can be transmitted by different types of plant material. Effective quarantine measures require specific, sensitive and rapid methods to detect Xap in propagative material or new reservoirs. Laborious and time-consuming methods for the diagnosis of Xap are recommended in the current EPPO standard protocol. However, new several pathogen-specific PCR and quantitative realtime PCR assays have been developed that enable direct detection of Xap in symptomatic and symptomless plant samples. A concise resource of current methods for Xap detection and identification, based on assessment and development activities within the framework of COST 873, is presented

Current status of bacterial spot of stone fruits and almond caused by Xanthomonas arboricola pv. pruni in Spain

In 2002, typical symptoms of bacterial spot disease of stone fruits caused by Xanthomomas arboricola pv. pruni (Xap) were observed for the first time on Japanese plum in Badajoz (south-western Spain). During the following years, the pathogen was found in seven other eastern and northern Spanish provinces (Valencia, Alicante, Zaragoza, Huesca, Navarra, Lérida and Mallorca) affecting different cultivars of Japanese plum, nectarine, peach and almond. There are few previous reports of Xap on almond, the Spanish outbreaks constituting its first detection on this host in the European Union (EU). Identification of the pathogen was performed using biochemical tests, fatty acid methyl esters (FAME) profiles, conventional and real-time PCR, and hypersensitivity reaction on tobacco leaves. Pathogenicity was demonstrated by inoculation of young potted plants of peach, plum or almond and successful re-isolations from plants with symptoms. In areas where infected plants were found, eradication programs were set up since Xap has a quarantine status according to phytosanitary EU legislation.Publishe

Evaluation of the efficiency of a conventional PCR protocol for the diagnosis of bacterial spot disease caused by Xanthomonas arboricola pv. pruni in stone fruits and almond

Xanthomonas arboricola pv. pruni (Xap), the causal agent of bacterial spot disease of stone fruits and almond, has a quarantine status for the European Union and the European and Mediterranean Plant Protection Organization. The symptoms in the diverse hosts show some differences and, although being quite typical, could be confused with those of some fungal diseases or other biotic or abiotic causes. Consequently, an accurate molecular diagnosis method is required for a rapid identification of the pathogen in samples of imported plants, from nurseries, orchards, etc. A protocol for conventional PCR designed by Pagani (2004) has been the only molecular analytic tool available for several years. It has been optimised for improving its specificity and sensitivity, and the results of its evaluation in 316 bacterial spot-like symptomatic samples of almond, apricot, cherry, Japanese plum and peach, compared with those of isolation and real-time PCR, are reported. The optimised PCR protocol showed specificity for a collection of Xap strains tested. Few non-desired reactions were obtained with some other xanthomonads which have not been reported from Prunus species. Sensitivity thresholds ranged from 102 to 105 CFU ml-1, depending on the hosts and type of plant material. This conventional PCR assay proved to be an excellent candidate for a rapid screening and presumptive diagnosis in cases where real-time PCR equipment is not available

Capacidad de supervivencia y transmisión por semilla de Xanthomonas arboricola pv. pruni en almendro

PublishedTrabajo financiado por el Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria, proyecto RTA2011-00140-d03-0

Efficiency of the EPPO protocol for preventing the introduction and dissemination of fire blight in two Spanish areas

COST-Action 864: PomeFruitHealth. Combining traditional and advanced strategies for plant protection in pome fruit growin

Dietary α‐Linolenic Acid, Marine ω‐3 Fatty Acids, and Mortality in a Population With High Fish Consumption: Findings From the PREvención con DIeta MEDiterránea (PREDIMED) Study

Background: Epidemiological evidence suggests a cardioprotective role of α‐linolenic acid (ALA), a plant‐derived ω‐3 fatty acid. It is unclear whether ALA is beneficial in a background of high marine ω‐3 fatty acids (long‐chain n‐3 polyunsaturated fatty acids) intake. In persons at high cardiovascular risk from Spain, a country in which fish consumption is customarily high, we investigated whether meeting the International Society for the Study of Fatty Acids and Lipids recommendation for dietary ALA (0.7% of total energy) at baseline was related to all‐cause and cardiovascular disease mortality. We also examined the effect of meeting the society's recommendation for long‐chain n‐3 polyunsaturated fatty acids (≥500 mg/day). Methods and Results: We longitudinally evaluated 7202 participants in the PREvención con DIeta MEDiterránea (PREDIMED) trial. Multivariable‐adjusted Cox regression models were fitted to estimate hazard ratios. ALA intake correlated to walnut consumption (r=0.94). During a 5.9‐y follow‐up, 431 deaths occurred (104 cardiovascular disease, 55 coronary heart disease, 32 sudden cardiac death, 25 stroke). The hazard ratios for meeting ALA recommendation (n=1615, 22.4%) were 0.72 (95% CI 0.56–0.92) for all‐cause mortality and 0.95 (95% CI 0.58–1.57) for fatal cardiovascular disease. The hazard ratios for meeting the recommendation for long‐chain n‐3 polyunsaturated fatty acids (n=5452, 75.7%) were 0.84 (95% CI 0.67–1.05) for all‐cause mortality, 0.61 (95% CI 0.39–0.96) for fatal cardiovascular disease, 0.54 (95% CI 0.29–0.99) for fatal coronary heart disease, and 0.49 (95% CI 0.22–1.01) for sudden cardiac death. The highest reduction in all‐cause mortality occurred in participants meeting both recommendations (hazard ratio 0.63 [95% CI 0.45–0.87]). Conclusions: In participants without prior cardiovascular disease and high fish consumption, dietary ALA, supplied mainly by walnuts and olive oil, relates inversely to all‐cause mortality, whereas protection from cardiac mortality is limited to fish‐derived long‐chain n‐3 polyunsaturated fatty acids. Clinical Trial Registration URL: http://www.Controlled-trials.com/. Unique identifier: ISRCTN35739639

Pseudomonas syringae pv. mori (Boyer y Lambert) Young, Dye, Wilkie

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