Variation and Heritability of Phenology in the Fungus Monilinia vaccinii-corymbosi on Blueberry

The germination of field-collected pseudosclerotia and the development of apothecia from eight New Jersey populations of the mummy berry fungus Monilinia vaccinii-corymbosi were evaluated under controlled conditions in the greenhouse. Development data for apothecia were used to describe the timing of apothecium formation and to estimate broad- and narrow-sense heritabilities of fungal phenology. Mean development times for the formation of apothecia ranged from 35.4 to 54.7 days. The mean development times for populations collected from early-season cv. Weymouth ranged from 35.4 to 39.6 days and were significantly shorter than the development times for three of the four populations collected from late-season cv. Jersey (46.9 to 54.7 days) or for the population collected from mixed stands of cultivated blueberries (42.7 days). The development of populations from late cultivars planted in very close proximity to early cv. Weymouth was early (36.5 to 39.0 days) and not significantly different from the development of populations collected from cv. Weymouth. Phenotypic and genetic variances of apothecium development for individual populations ranged from 18.9 to 44.8 and 7.2 to 30.9, respectively. Broad-sense heritabilities of apothecia development for each fungal population, calculated by partitioning phenotypic variation into genetic and environmental components, ranged from 0.31 to 0.78. Narrow-sense heritabilities of apothecia development, based on parent-offspring regression, ranged from 0.58 to 0.78. These results indicate that populations ofM. vaccinii-corymbosi differ in phenology and that a significant portion of the phenological variation within populations is genetic. Thus, it is plausible to propose that the phenology of apothecium development is a component of fungal fitness and that host phenology can influence the timing of pathogen development

Phenology of Apothecium Production in Populations of Monilinia vaccinii-corymbosi from Early- and Late-Maturing Blueberry Cultivars

Pseudosclerotia were evaluated for differences in timing of apothecium development in four controlled experiments conducted over a 2-year period. In a separate experiment, conidia from 10 randomly selected isolates from both of the fungal populations were used to inoculate open flowers. Germination of pseudosclerotia produced from these artificial inoculations also was evaluated. The timing and rate of shoot elongation for cvs. Weymouth and Jersey were assessed in one greenhouse and two field experiments. Average development times for the fungal population from cv. Weymouth were 8 to 15 days earlier or 33 to 42% less than those for the population from cv. Jersey. The fungal population from Weymouth also exhibited less variation in development times for each developmental stage measured. Similarly, germination of pseudosclerotia produced in artificial inoculations differed between populations. On average, pseudosclerotia derived from the Weymouth population produced apothecia 16 days earlier. During spring 1995 and 1996, vegetative and truss buds on cv. Weymouth developed 4 to 16 days earlier than those on cv. Jersey. These results demonstrate that M. vaccinii-corymbosi exhibits variation in timing of pseudosclerotia germination and apothecium development within and between populations. We hypothesize that differences observed in the timing of apothecium development are related to the fitness of the populations on their original host cultivars and were selected by host phenolog

Host Resistance to Monilinia vaccinii-corymbosi in Flowers and Fruits of Highbush Blueberry

Monilinia vaccinii-corymbosi, the causal agent of mummy berry disease, infects blueberry flowers via the gynoecial pathway. To describe the expression of host resistance in highbush blueberry (Vaccinium corymbosum), fungal growth in the styles and colonization of the locules were compared among five blueberry cultivars in a series of controlled greenhouse experiments. Styles were harvested 1 and 4 days postinoculation, and the length colonized by hyphae was determined using fluorescence microscopy. At 8 weeks after inoculation, fruit were harvested and scored for the presence of hyphae in the locules. The infection frequency of styles ranged from 0.33 to 0.71, and only cv. Weymouth had significantly lower infection frequency than the other cultivars. The mean length of the colonized portion of the stylar canal ranged from 0.126 to 0.434 mm after 1 day and 1.62 to 3.59 mm after 4 days. Hyphae in the styles of cv. Weymouth exhibited the least growth, whereas hyphae in the styles of cultivars Jersey and Rancocas were significantly longer. The distance of style penetrated for cultivars Bluecrop and Coville was intermediate. The mean disease incidence of locules differed significantly. Values for cultivars Weymouth and Jersey were the smallest (0.038 and 0.039) and largest (0.249 and 0.236), respectively. The results demonstrate that a component of resistance to infection by M. vaccinii-corymbosi is expressed during growth in the gynoecial pathway

Use of the tetrazolium salt MTT to measure cell viability effects of the bacterial antagonist Lysobacter enzymogenes on the filamentous fungus Cryphonectria parasitica

Despite substantial interest investigating bacterial mechanisms of fungal growth inhibition, there are few methods available that quantify fungal cell death during direct interactions with bacteria. Here we describe an in vitro cell suspension assay using the tetrazolium salt MTT as a viability stain to assess direct effects of the bacterial antagonist Lysobacter enzymogenes on hyphal cells of the filamentous fungus Cryphonectria parasitica. The effects of bacterial cell density, fungal age and the physiological state of fungal mycelia on fungal cell viability were evaluated. As expected, increased bacterial cell density correlated with reduced fungal cell viability over time. Bacterial effects on fungal cell viability were influenced by both age and physiological state of the fungal mycelium. Cells obtained from 1-week-old mycelia lost viability faster compared with those from 2-week-old mycelia. Likewise, hyphal cells obtained from the lower layer of the mycelial pellicle lost viability more quickly compared with cells from the upper layer of the mycelial pellicle. Fungal cell viability was compared between interactions with L. enzymogenes wildtype strain C3 and a mutant strain, DCA, which was previously demonstrated to lack in vitro antifungal activity. Addition of antibiotics eliminated contributions to MTT-formazan production by bacterial cells, but not by fungal cells, demonstrating that mutant strain DCA had lost complete capacity to reduce fungal cell viability. These results indicate this cell suspension assay can be used to quantify bacterial effects on fungal cells, thus providing a reliable method to differentiate strains during bacterial/fungal interactions

Population Dynamics and Genetics of Plant Disease: A Case Study of Anther-Smut Disease

This is the publisher's version, also available electronically from http://www.jstor.org/stable/info/2265569A model by Levin and Udovic (1977) emphasizes the need for integration of studies of the numerical abundances and genetic composition of host and pathogen species. We use their conceptual framework to summarize our collaborative research on the ecological genetics of the anther-smut disease of Silene alba caused by the fungus Ustilago violacea. Our theoretical investigations have revealed the importance of the rate and mode of disease transmission on the likelihood of coexistence between host and pathogen. Our empirical studies have quantified patterns of disease spread on local and regional spatial scales and have shown that host genotypes differ greatly in resistance. Comparable genetic variation in pathogen virulence has not yet been demonstrated. The genetic composition of host populations alters numerical dynamics in experimental populations: disease declines in resistant populations, while host and pathogen appear to coexist in susceptible populations. Ecological outcomes also may be affected by the negative relationship between disease incidence and host flowering time, which may constitute a "cost" to resistance. We are currently expanding our work on the ecology and genetics of metapopulation dynamics of host and pathogen

The Distribution of Mating-Type Bias in Natural Populations of the Anther-Smut Ustilago violacea on Silene alba in Virginia

This is the publisher's version, also available electronically from http://www.jstor.org/stable/info/3761395.Complete individual-wide mating-type bias (retrieval of sporidia of only one mating type from germinated teliospores of one fungal individual) was observed to be a common and widespread feature of the anther-smut fungus, Ustilago violacea, collected from natural populations of its host, Silene alba. The bias was usually to mating type A1, but the frequency of bias and its spatial distribution varied from region to region. Populations with high frequencies of bias still showed high rates of disease transmission. Crosses between A1 mating type sporidial lines from completely biased individuals and A2 mating types from unbiased individuals showed no bias in the progeny. During teliospore germination, biased individuals often showed conjugation among adjacent cells of the promycelium, suggesting that both mating types are present in the germinating teliospore but one mating type is unable to grow as free-living sporidia. The complete bias was most readily interpreted as evidence of "haploid lethals" linked to mating type that cause poor survival or growth of the sporidial stage. The results show that such "haploid lethals" may be a common occurrence in natural populations, and that fungal mating systems may vary considerably over short distances

Assessment and comparison of rhizosphere communities in cultivated Vaccinium spp. provide a baseline for study of causative agents in decline

It has long been recognized that the community of organisms associated with plant roots is a critical component of the phytobiome and can directly or indirectly contribute to the overall health of the plant. The rhizosphere microbial community is influenced by a number of factors including the soil type, the species of plants growing in those soils, and in the case of cultivated plants, the management practices associated with crop production. Vaccinium species, such as highbush blueberry and American cranberry, are woody perennials that grow in sandy, acidic soils with low to moderate levels of organic matter and a paucity of nutrients. When properly maintained, fields planted with these crops remain productive for many years. In some cases, however, yields and fruit quality decline over time, and it is suspected that degenerating soil health and/or changes in the rhizosphere microbiome are contributing factors. Determining the assemblage of bacterial and fungal microorganisms typically associated with the rhizosphere of these crops is a critical first step toward addressing the complex issue of soil health. We hypothesized that since blueberry and cranberry are in the same genus and grow in similar soils, that their associated rhizosphere microbial communities would be similar to each other. We analyzed the eukaryotic (primarily fungal) and bacterial communities from the rhizosphere of representative blueberry and cranberry plants growing in commercial fields in New Jersey. The data presented herein show that while the bacterial communities between the crops is very similar, the fungal communities associated with each crop are quite different. These results provide a framework for examining microbial components that might contribute to the health of Vaccinium spp. crops in New Jersey and other parts of the northeastern U.S

Habitat and Host Indicate Lineage Identity in Colletotrichum gloeosporioides s.l. from Wild and Agricultural Landscapes in North America

Understanding the factors that drive the evolution of pathogenic fungi is central to revealing the mechanisms of virulence and host preference, as well as developing effective disease control measures. Prerequisite to these pursuits is the accurate delimitation of species boundaries. Colletotrichum gloeosporioides s.l. is a species complex of plant pathogens and endophytic fungi for which reliable species recognition has only recently become possible through a multi-locus phylogenetic approach. By adopting an intensive regional sampling strategy encompassing multiple hosts within and beyond agricultural zones associated with cranberry (Vaccinium macrocarpon Aiton), we have integrated North America strains of Colletotrichum gloeosporioides s.l. from these habitats into a broader phylogenetic framework. We delimit species on the basis of genealogical concordance phylogenetic species recognition (GCPSR) and quantitatively assess the monophyly of delimited species at each of four nuclear loci and in the combined data set with the genealogical sorting index (gsi). Our analysis resolved two principal lineages within the species complex. Strains isolated from cranberry and sympatric host plants are distributed across both of these lineages and belong to seven distinct species or terminal clades. Strains isolated from V. macrocarpon in commercial cranberry beds belong to four species, three of which are described here as new. Another species, C. rhexiae Ellis & Everh., is epitypified. Intensive regional sampling has revealed a combination of factors, including the host species from which a strain has been isolated, the host organ of origin, and the habitat of the host species, as useful indicators of species identity in the sampled regions. We have identified three broadly distributed temperate species, C. fructivorum, C. rhexiae, and C. nupharicola, that could be useful for understanding the microevolutionary forces that may lead to species divergence in this important complex of endophytes and plant pathogens

Renal replacement therapy in acute kidney injury: controversy and consensus

Renal replacement therapies (RRTs) represent a cornerstone in the management of severe acute kidney injury. This area of intensive care and nephrology has undergone significant improvement and evolution in recent years. Continuous RRTs have been a major focus of new technological and treatment strategies. RRT is being used increasingly in the intensive care unit, not only for renal indications but also for other organ-supportive strategies. Several aspects related to RRT are now well established, but others remain controversial. In this review, we review the available RRT modalities, covering technical and clinical aspects. We discuss several controversial issues, provide some practical recommendations, and where possible suggest a research agenda for the future

Coronary Angiography After Cardiac Arrest Without ST Segment Elevation:One-Year Outcomes of the COACT Randomized Clinical Trial

Importance: Ischemic heart disease is a common cause of cardiac arrest. However, randomized data on long-term clinical outcomes of immediate coronary angiography and percutaneous coronary intervention (PCI) in patients successfully resuscitated from cardiac arrest in the absence of ST segment elevation myocardial infarction (STEMI) are lacking. Objective: To determine whether immediate coronary angiography improves clinical outcomes at 1 year in patients after cardiac arrest without signs of STEMI, compared with a delayed coronary angiography strategy. Design, Setting, and Participants: A prespecified analysis of a multicenter, open-label, randomized clinical trial evaluated 552 patients who were enrolled in 19 Dutch centers between January 8, 2015, and July 17, 2018. The study included patients who experienced out-of-hospital cardiac arrest with a shockable rhythm who were successfully resuscitated without signs of STEMI. Follow-up was performed at 1 year. Data were analyzed, using the intention-to-treat principle, between August 29 and October 10, 2019. Interventions: Immediate coronary angiography and PCI if indicated or coronary angiography and PCI if indicated, delayed until after neurologic recovery. Main Outcomes and Measures: Survival, myocardial infarction, revascularization, implantable cardiac defibrillator shock, quality of life, hospitalization for heart failure, and the composite of death or myocardial infarction or revascularization after 1 year. Results: At 1 year, data on 522 of 552 patients (94.6%) were available for analysis. Of these patients, 413 were men (79.1%); mean (SD) age was 65.4 (12.3) years. A total of 162 of 264 patients (61.4%) in the immediate angiography group and 165 of 258 patients (64.0%) in the delayed angiography group were alive (odds ratio, 0.90; 95% CI, 0.63-1.28). The composite end point of death, myocardial infarction, or repeated revascularization since the index hospitalization was met in 112 patients (42.9%) in the immediate group and 104 patients (40.6%) in the delayed group (odds ratio, 1.10; 95% CI, 0.77-1.56). No significant differences between the groups were observed for the other outcomes at 1-year follow-up. For example, the rate of ICD shocks was 20.4% in the immediate group and 16.2% in the delayed group (odds ratio, 1.32; 95% CI, 0.66-2.64). Conclusions and Relevance: In this trial of patients successfully resuscitated after out-of-hospital cardiac arrest and without signs of STEMI, a strategy of immediate angiography was not found to be superior to a strategy of delayed angiography with respect to clinical outcomes at 1 year. Coronary angiography in this patient group can therefore be delayed until after neurologic recovery without affecting outcomes