Gray leaf spot (GLS) is a destructive disease of perennial ryegrass caused by a host specific pathotype of the ascomycete Magnaporthe oryzae. Early diagnosis is crucial for effective disease management and the implementation of Integrated Pest Management practices. However, a rapid protocol for the detection of low levels of airborne inoculum is still missing. We developed a pathogen-specific quantitative loop-mediated isothermal amplification (qLAMP) assay coupled with a spore trap system for rapid detection and quantification of airborne inoculum of the M. oryzae perennial ryegrass pathotype, and tested its suitability for implementation in GLS-infected turfgrass fields. In summer 2015, two perennial ryegrass plots were artificially inoculated with the pathogen, with four continuously running custom impaction spore traps placed in each plot. Sampling units were replaced daily and tested with the developed qLAMP assay, while plots were monitored for symptom development. Results confirmed that the qLAMP assay-trap system was able to detect as few as 10 conidia up to 12 days before symptoms developed in the field. LAMP technology is particularly appropriate for field implementation by nontechnical users, and has the potential to be a powerful decision support tool to guide timing of fungicide applications for GLS management.

Field surveys conducted throughout California olive-growing regions in 2008 and 2009 resulted in a collection of 101 Cytospora-like isolates from olive twig and branch dieback symptoms. Cytospora isolates were isolated from multiple cvs. in different olive orchards in Fresno, Madera, Merced, Napa, Riverside, Santa Barbara, Sonoma, Tulare, and Ventura counties. Taxonomic studies of macro- and microscopic structures along with multigene phylogenetic analyses of the internal transcribed spacer region, including the 5.8S rDNA (ITS1–5.8S–ITS2), and fragments of the translation elongation factor 1-α, beta-tubulin, and actin genes identified two species, Cytospora oleicola and C. olivarum sp. nov. Pathogenicity studies conducted in mature olive trees cvs. Manzanillo and Sevillano showed both species to be pathogenic and able to cause vascular necrosis and cankers in olive branches. This study adds to the current knowledge on the etiology of olive twig and branch dieback and provides new important information for the development of effective control strategies against canker diseases affecting olive in California.

Boxwood blight caused by Calonectria pseudonaviculata (Cps) and C. henricotiae (Che) is destroying cultivated and native boxwood worldwide, with profound negative economic impacts on the horticulture industry. First documented in the U.S. in 2011, the disease has now occurred in 28 states. Previous research showed that global Cps populations prior to 2014 had a clonal structure, and only the MAT1-2 idiomorph was observed. In this study, we examined the Cps genetic diversity and population structure in the U.S. after 2014, following the expansion of the disease across the country over the past five years. Two hundred eighteen isolates from 21 states were genotyped by sequencing 11 simple sequence repeat (SSR) loci and by MAT1 idiomorph typing. All isolates presented Cps-specific alleles, indicating that Che is still absent in the U.S. states sampled. The presence of only the MAT1-2 idiomorph and gametic linkage disequilibrium suggests the prevalence of asexual reproduction. The contemporary Cps population is characterized by a clonal structure and composed of 13 multilocus genotypes (SSR-MLGs) unevenly distributed across the U.S. These SSR-MLGs grouped into two clonal lineages (CL). The predominant lineage CL2 (93% of isolates) is the primary contributor to U.S. disease expansion. The contemporary U.S. Cps population is not geographically subdivided and not genetically differentiated from the U.S. population prior to 2014, but is significantly differentiated from the main European population, which is largely composed of CL1. Our findings provide insights into the boxwood blight epidemic that are critical for disease management and breeding of resistant boxwood cultivars.

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This study investigates the diversity and taxonomy of Cryptosphaeria species occurring in the western United States on the basis of morphological characters and multilocus phylogenetic analyses of the ribosomal internal transcribed spacer region, parts of a β-tubulin gene, the DNA-dependent RNA polymerase II second-largest subunit gene and the nuclear ribosomal large subunit gene. Cryptosphaeria multicontinentalis sp. nov is described from the Sierra Nevada and central coast of California on Populus tremuloides, P. balsamifera subsp. trichocarpa and P. fremontii. Cryptosphaeria pullmanensis is reported from a wide geographic area in the western United States on the main host, P. fremontii. The pathogen C. lignyota is reported for the first time from the Sierra Nevada of California on P. tremuloides. The phylogenetic analyses showed that C. multicontinentalis is a sister species to C. lignyota. Both species were closely related to C. subcutanea and more distantly related to C. pullmanensis. Characteristics of both teleomorph and anamorph of the newly introduced species C. multicontinentalis are described and illustrated.