Effect of Temperature, Leaf Wetness Period, and Cultivar Susceptibility on Boxwood Blight Disease Development and Sporulation.

Boxwood blight causes great losses to the boxwood nursery industry and landscapes in 30 states in the United States. Understanding the epidemiological factors governing disease development will be important for disease forecasting and design of best management practices. We evaluated the effect of leaf wetness period (lwp) and temperature on lesion development and sporulation on three boxwood cultivars under controlled conditions to develop predictive models for disease development. We conducted detached leaf assays at 18 to 27°C and various lwp with the cultivars Buxus sempervirens 'Suffruticosa' (highly susceptible), B. sempervirens × B. microphylla var. koreana 'Green Velvet' (moderately susceptible), and B. microphylla var. japonica 'Winter Gem' (less susceptible). Detached leaves were inoculated with 200 conidia in 50 µl of suspension and disease incidence was recorded at 3 to 13 days postinoculation (dpi). Cultivar, lwp, temperature, and most interactions significantly influenced disease development. A minimum of 5 h of leaf wetness was required for any disease. Lesion development increased most rapidly between 12 and 15 h and continued to increase to about 21 h of leaf wetness. Temperatures between 21 and 25°C were optimal for lesion development. There was about a 7-day lag between appearance of lesions and maximal incidence of sporulation. The two less-susceptible cultivars had fewer lesions than Suffruticosa under the same infection conditions; in addition, leaf lesions of Winter Gem exhibited delayed sporulation and sporulation from a smaller proportion of symptomatic leaves. Response surfaces were developed for each cultivar to predict the disease incidence using the lwp and dpi. Our findings will help refine disease forecast models to improve management of boxwood blight.

Mosaic fungal individuals have the potential to evolve within a single generation.

Although cells of mushroom-producing fungi typically contain paired haploid nuclei (n + n), most Armillaria gallica vegetative cells are uninucleate. As vegetative nuclei are produced by fusions of paired haploid nuclei, they are thought to be diploid (2n). Here we report finding haploid vegetative nuclei in A. gallica at multiple sites in southeastern Massachusetts, USA. Sequencing multiple clones of a single-copy gene isolated from single hyphal filaments revealed nuclear heterogeneity both among and within hyphae. Cytoplasmic bridges connected hyphae in field-collected and cultured samples, and we propose nuclear migration through bridges maintains this nuclear heterogeneity. Growth studies demonstrate among- and within-hypha phenotypic variation for growth in response to gallic acid, a plant-produced antifungal compound. The existence of both genetic and phenotypic variation within vegetative hyphae suggests that fungal individuals have the potential to evolve within a single generation in response to environmental variation over time and space.

DESS deconstructed: Is EDTA solely responsible for protection of high molecular weight DNA in this common tissue preservative?

DESS is a formulation widely used to preserve DNA in biological tissue samples. Although it contains three ingredients, dimethyl sulfoxide (DMSO), ethylenediaminetetraacetic acid (EDTA) and sodium chloride (NaCl), it is frequently referred to as a DMSO-based preservative. The effectiveness of DESS has been confirmed for a variety of taxa and tissues, however, to our knowledge, the contributions of each component of DESS to DNA preservation have not been evaluated. To address this question, we stored tissues of three aquatic taxa, Mytilus edulis (blue mussel), Faxonius virilis (virile crayfish) and Alitta virens (clam worm) in DESS, each component of DESS individually and solutions containing all combinations of two components of DESS. After storage at room temperature for intervals ranging from one day to six months, we extracted DNA from each tissue and measured the percentage of high molecular weight (HMW) DNA recovered (%R) and normalized HMW DNA yield (nY). Here, HMW DNA is defined as fragments >10 kb. For comparison, we also measured the %R and nY of HMW DNA from extracts of fresh tissues and those stored in 95% EtOH over the same time intervals. We found that in cases where DESS performed most effectively (yielding ≥ 20%R of HMW DNA), all solutions containing EDTA were as or more effective than DESS. Conversely, in cases where DESS performed more poorly, none of the six DESS-variant storage solutions provided better protection of HMW DNA than DESS. Moreover, for all taxa and storage intervals longer than one day, tissues stored in solutions containing DMSO alone, NaCl alone or DMSO and NaCl in combination resulted in %R and nY of HMW DNA significantly lower than those of fresh tissues. These results indicate that for the taxa, solutions and time intervals examined, only EDTA contributed directly to preservation of high molecular weight DNA.

The Identification of a New Species, Diaporthe humulicola, a Pathogen Causing Diaporthe Leaf Spot on Common Hop.

Common hop, Humulus lupulus, is a commercially important crop in the United States, with an increasing number of hop yards being established in the Northeast. In 2018, a new fungal disease was observed at two research hop yards in Connecticut. This new pathogen affected all hop cultivars being grown and caused leaf spots and browning of cones. The causal organism was isolated and Koch's postulates were performed to confirm pathogenicity. The disease symptoms were similar to the previously described Phoma wilt; however, morphological and phylogenetic analyses placed the causal organism as a new species of Diaporthe. We propose the name Diaporthe humulicola. The disease increased under hot, humid conditions (around 24°C and 90% relative humidity), which prevail during the summer in the northeastern United States as well as other parts of the country. An in vitro preliminary assessment of fungicide sensitivity revealed that pyraclostrobin and boscalid inhibited D. humulicola growth in culture and should be further assessed for field efficacy against this new disease of hop. The proper identification and monitoring of this pathogen will be important to inform hop growers of this new threat.

Draft Genome Sequences of the Turfgrass Pathogen Sclerotinia homoeocarpa.

Sclerotinia homoeocarpa (F. T. Bennett) is one of the most economically important pathogens on high-amenity cool-season turfgrasses, where it causes dollar spot. To understand the genetic mechanisms of fungicide resistance, which has become highly prevalent, the whole genomes of two isolates with varied resistance levels to fungicides were sequenced.