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Needle blights are serious needle fungal diseases affecting pines both in natural and productive forests. Among needle blight agents, the ascomycetes Lecanosticta acicola, Dothistroma pini and D. septosporum are of particular concern. These pathogens need specific, fast and accurate diagnostics since they are regulated species in many countries and may require differential management measures. Due to the similarities in fungal morphology and the symptoms they elicit, these species are hard to distinguish using morphological characteristics. The symptoms can also be confused with those caused by insects or abiotic agents. DNA-based detection is therefore recommended. However, the specific PCR assays that have been produced to date for the differential diagnosis of these pathogens can be applied only in a well-furnished laboratory and the procedure takes a relatively long execution time. Surveillance and forest protection would benefit from a faster diagnostic method, such as a loop-mediated isothermal amplification (LAMP) assay, which requires less sophisticated equipment and can also be deployed directly on-site using portable devices. LAMP assays for the rapid and early detection of L. acicola, D. pini and D. septosporum were developed in this work. Species-specific LAMP primers and fluorescent assimilating probes were designed for each assay, targeting the beta tubulin (β-tub2) gene for the two Dothistroma species and the elongation factor (EF-1α) region for L. acicola. Each reaction detected its respective pathogen rapidly and with high specificity and sensitivity in DNA extracts from both pure fungal cultures and directly from infected pine needles. These qualities and the compatibility with inexpensive portable instrumentation position these LAMP assays as an effective method for routine phytosanitary control of plant material in real time, and they could profitably assist the management of L. acicola, D. pini and D. septosporum. Full article

Red band needle blight caused by Dothistroma septosporum and D. pini, and brown spot needle blight caused by Lecanosticta acicola provoke severe and premature defoliation in Pinus, and subsequent reduction of photosynthetic surfaces, vitality, and growth in young and adult trees. The recurrent damage results in branch and tree death. Until recently, pine needle blight diseases have had only minor impacts on native and exotic forest trees in the North of Spain, but in the past five years, these pathogen species have spread widely and caused severe defoliation and mortality in exotic and native plantations of Pinus in locations where they were not detected before. In an attempt to understand the main causes of this outbreak and to define the effectiveness of owners’ management strategies, four research actions were implemented: a survey of the management activities implemented by the owners to reduce disease impact, the evaluation of specific symptoms and damage associated with infection, and the identification of the causative pathogenic species and their reproductive capacity. Morphological characteristics of the fungus and molecular identification were consistent with those of Lecanosticta acicola and Dothistroma spp., D. septosporum, D. Pini, and both mating types were present for the three identified pathogens. The local silvicultural management performed, mainly pruning and thinning, was not resulting in the expected improvement. The results of this study can be applied to establish guidelines for monitoring and controlling the spread of needle blight pathogens. Full article

Dothistromin is a non-host selective toxin produced by the pine needle pathogen Dothistroma septosporum. Dothistromin is not required for pathogenicity, but may have a role in competition and niche protection. To determine how D. septosporum tolerates its own toxin, a putative dothistromin transporter, DotC, was investigated. Studies with mutants lacking a functional dotC gene, overproducing DotC, or with a DotC-GFP fusion gene, did not provide conclusive evidence of a role in dothistromin efflux. The mutants revealed a major effect of DotC on dothistromin biosynthesis but were resistant to exogenous dothistromin. Intracellular localization studies suggest that compartmentalization may be important for dothistromin tolerance. Full article