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Calonectria pseudonaviculata (Cps) infects Buxus (boxwood), Pachysandra (pachysandra), and Sarcococca spp. (sweet box); yet, how it adapts to its hosts has been unclear. Here, we performed serial passage experiments with the three hosts and measured Cps changes in three aggressiveness components: infectibility, lesion size, and conidial production. The detached leaves of individual hosts were inoculated with isolates (P0) from the originating host, followed by nine serial inoculations of new leaves of the same host with conidia from the infected leaves of the previous inoculation. All boxwood isolates maintained their capability of infection and lesion expansion through the 10 passages, whereas most non-boxwood isolates lost these abilities during the passages. Isolates from plants of origin (*-P0) and their descendants isolated from passages 5 (*-P5) and 10 (*-P10) were used to evaluate aggressiveness changes on all three hosts with cross-inoculation. While post-passage boxwood isolates gave enlarged lesions on pachysandra, sweet box P5 and pachysandra P10 isolates showed reduced aggressiveness on all hosts. Cps appears to be most adapted to boxwood and less adapted to sweet box and pachysandra. These results suggest speciation of Cps, with its coevolutionary pace with the hosts the fastest with boxwood, intermediate with sweet box, and the slowest with pachysandra.

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Impatiens downy mildew (IDM) caused by Plasmopara destructor is currently the primary constraint on the production and use of impatiens (Impatiens walleriana) as bedding plants worldwide. Downy mildew has been documented since the 1880s from wild-grown Impatiens spp. but epidemic outbreaks of the disease affecting the commercially grown, ornamental I. walleriana were only reported for the first time in 2003 in the United Kingdom and in 2004 in the United States. Here, we assess the genetic diversity, level of differentiation, and population structure from 623 samples associated with current and preepidemic IDM outbreaks, by genotyping the samples with simple sequence repeat markers. P. destructor population structure following the emergence of IDM in the United States is subdivided into four genetic lineages characterized by high genetic diversity, mixed reproduction mode, inbreeding, and an excess of heterozygosity. P. destructor genotypes are significantly differentiated from preepidemic IDM samples from hosts other than I. walleriana but no geographical or temporal subdivision is evident. P. destructor samples from different Impatiens spp. show significant but very low levels of differentiation in the analysis of molecular variance test that did not hold in discriminant analysis of principal components analyses. The same was observed between samples of P. destructor and P. velutina recovered from I. walleriana. The finding of shared genotypes in samples from different countries and lack of differentiation among U.S. and Costa Rican samples indicate the occurrence of international movement of the pathogen. Our study provides the first high-resolution analysis of the diversity of P. destructor populations and the IDM epidemic that may be instrumental for disease management and breeding efforts.

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Globalization has made agricultural commodities more accessible, available, and affordable. However, their global movement increases the potential for invasion by pathogens and necessitates development and implementation of sensitive, rapid, and scalable surveillance methods. Here, we used 35 strains, isolated by multiple diagnostic laboratories, as a case study for using whole genome sequence data in a plant disease diagnostic setting. Twenty-seven of the strains were isolated in 2022 and identified as Xanthomonas hortorum pv. pelargonii. Eighteen of these strains originated from material sold by a plant breeding company that had notified clients following a release of infected geranium cuttings. Analyses of whole genome sequences revealed epidemiological links among the 27 strains from different growers that confirmed a common source of the outbreak and uncovered likely secondary spread events within facilities that housed plants originating from different plant breeding companies. Whole genome sequencing data were also analyzed to reveal how preparatory and analytical methods can impact conclusions on outbreaks of clonal pathogenic strains. The results demonstrate the potential power of using whole genome sequencing among a network of diagnostic labs and highlight how sharing such data can help shorten response times to mitigate outbreaks more expediently and precisely than standard methods.

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Boxwood blight caused by Calonectria pseudonaviculata and C. henricotiae is destroying cultivated and native boxwood worldwide, with profound negative economic impacts on the horticulture industry. First documented in the United States in 2011, the disease has now occurred in 30 states. Previous research showed that global C. pseudonaviculata populations prior to 2014 had a clonal structure, and only the MAT1-2 idiomorph was observed. In this study, we examined C. pseudonaviculata genetic diversity and population structure in the United States after 2014, following the expansion of the disease across the country over the past 5 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 C. pseudonaviculata-specific alleles, indicating that C. henricotiae 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 C. pseudonaviculata population is characterized by a clonal structure and composed of 13 multilocus genotypes (SSR-MLGs) unevenly distributed across the United States. These SSR-MLGs grouped into two clonal lineages (CLs). The predominant lineage CL2 (93% of isolates) is the primary contributor to U.S. disease expansion. The contemporary U.S. C. pseudonaviculata 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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Ornamental plants in the genus Phlox are extensively planted in landscapes and home gardens around the world. A major limitation to a more widespread use of these plants is their susceptibility to powdery mildew (PM). In this study, we used multilocus sequence typing (MLST) analysis to gain insights into the population diversity of 32 Phlox PM pathogen (Golovinomyces magnicellulatus and Podosphaera sp.) isolates collected from the eastern United States and relate it to the ability to overcome host resistance. Low genetic diversity and a lack of structure were found within our population. Whole genome comparison of two isolates was used to support low genetic diversity evidence found with the MLST analysis. Recombination was suggested by the incongruences observed in the six phylogenetic trees generated from the housekeeping genes TEF-1α, CSI, ITS, IGS, H3, and TUB. Contrasting with low genetic diversity, we found high phenotypic diversity when using 10 of the 32 isolates to evaluate host resistance in four different Phlox species (P. paniculata 'Dunbar Creek', P. amoena OPGC 3598, P. glaberrima OPGC 3594, and P. subulata OPGC 4185) using in vitro bioassays. We observed quantitative and qualitative resistance in all Phlox species and a consistent low disease severity in our control, P. paniculata 'Dunbar Creek'. Taken together, the results generated in this study constitute a robust screening of popular Phlox germplasm that can be incorporated into breeding programs for PM resistance and provides significant information on the evolution of PM pathogens.

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Boxwood blight, caused by Calonectria pseudonaviculata and Calonectria henricotiae, has had devastating effects in gardens since its first appearance in the United Kingdom in 1994. The disease affects two other plants in the Buxaceae: sweet box (Sarcococca spp.) and pachysandra (Pachysandra spp.). C. pseudonaviculata was likely introduced to Europe by nursery trade from East Asia on an ornamental species and then to western Asia and North America. Thus far, C. henricotiae has been seen only in Europe. Boxwood, valued at $126 million wholesale per year in the United States alone, is now besieged by an aggressive foliar blight active over a broad temperature range when there are long periods of leaf wetness. Research on inoculum, means of dissemination, cultivar susceptibility, environmental influences, fungicides, sanitizers, and detection methods has vastly improved knowledge of this new invasive disease in a short time. Boxwood with genetic resistance to the disease is critically needed.

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Groundnut ringspot virus (GRSV) and Tomato chlorotic spot virus (TCSV) are two emerging tospoviruses in Florida. In a survey of the southeastern United States, GRSV and TCSV were frequently detected in solanaceous crops and weeds with tospovirus-like symptoms in south Florida, and occurred sympatrically with Tomato spotted wilt virus (TSWV) in tomato and pepper in south Florida. TSWV was the only tospovirus detected in other survey locations, with the exceptions of GRSV from tomato (Solanum lycopersicum) in South Carolina and New York, both of which are first reports. Impatiens (Impatiens walleriana) and lettuce (Lactuca sativa) were the only non-solanaceous GRSV and/or TCSV hosts identified in experimental host range studies. Little genetic diversity was observed in GRSV and TCSV sequences, likely due to the recent introductions of both viruses. All GRSV isolates characterized were reassortants with the TCSV M RNA. In laboratory transmission studies, Frankliniella schultzei was a more efficient vector of GRSV than F. occidentalis. TCSV was acquired more efficiently than GRSV by F. occidentalis but upon acquisition, transmission frequencies were similar. Further spread of GRSV and TCSV in the United States is possible and detection of mixed infections highlights the opportunity for additional reassortment of tospovirus genomic RNAs.

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Fungus gnats have been shown to transmit a variety of plant-pathogenic fungi that produce aerial dispersal stages. However, few studies have examined potential interactions between fungus gnats and oomycetes, including Pythium spp. A series of laboratory experiments were conducted to determine whether fungus gnat adults are vectors of several common greenhouse Pythium spp., including Pythium aphanidermatum, P. irregulare, and P. ultimum. An additional objective was to determine whether P. aphanidermatum can be maintained transstadially in the gut of a fungus gnat larva through the pupal stadium to be transmitted by the subsequent adult. Adult fungus gnats did not pick up infectious Pythium propagules from diseased plants and transmit them to healthy plants in any experiment. Species-specific primers and a probe for real-time polymerase chain reaction were developed to detect the presence of P. aphanidermatum DNA in fungus gnat tissue samples. P. aphanidermatum DNA was detectable in the larval and pupal stages; however, none was detected in adult fungus gnats. These results are in agreement with previous studies that have suggested that adult fungus gnats are unlikely vectors of Pythium spp.

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This is the first North American report of Oidium longipes, an anamorphic powdery mildew species described recently in Europe. It was found on vegetatively propagated petunia grown in a commercial greenhouse in New Jersey, USA, where it caused a rapidly spreading disease. The pathogen might have originated offshore and may have already been distributed in the United States through horticultural trade. During field surveys in Europe, it was found on petunia in Hungary and Austria as well; this is the first report of O. longipes from these two countries. A detailed light microscopy study of American and European specimens of O. longipes, including freshly collected samples and authentic herbarium specimens, revealed that its conidiophore morphology is more variable than illustrated in the original species description or in subsequent works. Microcycle conidiation, a process not yet known to occur in powdery mildews, was repeatedly observed in O. longipes. The rDNA internal transcribed spacer (ITS) sequences were identical in colonies containing different conidiophore types as well as in a total of five specimens collected from petunia in the United States, Austria, Hungary, Germany, and Switzerland. A phylogenetic analysis of the ITS sequences revealed that the closest known relative of O. longipes is O. lycopersici, known to infect tomato only in Australia. Cross-inoculation tests showed that O. longipes from petunia heavily infected tobacco cv. Xanthi, while the tomato and eggplant cultivars tested were moderately susceptible to this pathogen. These results indicate that its spread represents a potential danger to a number of solanaceous crops. Our ad hoc field surveys conducted in 2006 and 2007 did not detect it outside New Jersey in the United States; all the other powdery mildew-infected petunias, collected in New York and Indiana, were infected by Podosphaera xanthii. In Europe, most of the powdery mildew-infected petunias examined in this study were infected by P. xanthii or Golovinomyces orontii. Our multiple inoculation tests revealed that the same petunia plants and even the same leaves can be infected concomitantly by O. longipes, O. neolycopersici, G. orontii, and P. xanthii. Thus, it is at present unclear to what extent O. longipes contributes to the powdery mildew epidemics that develop year after year on solanaceous plants in many parts of the world.

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A filamentous virus identified in coleus (Coleus × hybrida) in Minnesota and New York was found to cause veinal necrosis in coleus, although this symptom was observed only under certain conditions. The virus was transmitted readily by mechanical inoculation to coleus and Nicotiana spp. and was not transmitted by Myzus persicae. The particles of the coleus virus had a modal length of 640 nm and a single capsid protein with an estimated molecular mass of 34 kDa. The amino acid sequence of the coat protein region of the coleus virus genome had significant similarities only to the corresponding domain of carlaviruses. Based on virion morphology, capsid protein size, genome size and organization, amino acid sequence, and phylogenetic analyses, the coleus virus, which was named provisionally Coleus vein necrosis virus (CVNV), was concluded to be a new definitive member of the genus Carlavirus. A 2-kb fragment of the 3' terminus of the CVNV genome sequence is accessible under accession number DQ915963 in GenBank.

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Economic, environmental, and technological influences complicate the task of achieving disease-free products in the ornamentals industry. Integrated pest management (IPM) is a cornerstone of floriculture and nursery crop production: strategies include sanitation, clean stock, host resistance, and control through biological, cultural, environmental, chemical, and regulatory means. Sanitation measures and cultural controls must keep pace with new production technologies. Clean stock programs are used for many crops that are propagated vegetatively. Breeding, selection, and biotechnology provide crops resistant to pathogens. Offshore production for economic competitiveness can introduce pathogens that make regulatory programs necessary. New biocontrol and chemical products continue to improve control while meeting the requirement for minimal environmental impact. Continual introduction of new crops and new production technologies creates new opportunities for pathogens to exploit, such that new disease management tactics must be discovered and old ones rediscovered to achieve optimum health management for ornamentals.

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Since 1997, powdery mildew infections have been repeatedly observed on Sedum spectabile plants, cv. Autumn Joy, grown as ornamentals in commercial greenhouses in New York. Circular patches of gray mycelia appeared and spread on upper and occasionally on lower leaf surfaces followed by necrosis of the leaf tissues and defoliation. The new disease reduced the market value of the infected ornamentals and required chemical control. The pathogen produced conidia singly on 2- to 3-celled conidiophores occurring on the ectophytic hyphae. Conidia were subcylindrical, measured 27 to 36 µm × 13 to 17 µm, and contained no fibrosin bodies. Germinating conidia produced a short germ tube, 5 to 30 µm, terminating in a lobed appressorium. Hyphal appressoria were lobed to multi-lobed, opposite or spread along the hyphae. Cleistothecia were not found. Based on conidial characteristics, the pathogen was identified as Erysiphe sedi Braun. To confirm pathogenicity, potted healthy S. spectabile plants were placed near infected plants in the greenhouse. In addition, detached S. spectabile leaves were inoculated with the pathogen by touching them to powdery mildew colonies and then placed in plates filled with one layer of polystyrene balls floated in water. Plates were covered and kept in the laboratory. Uninfected potted plants kept in another greenhouse and noninoculated detached leaves served as controls. After 1 week, powdery mildew appeared on all infected plants and leaves exposed to or inoculated with the pathogen. The pathogen was morphologically identical to the original fungus. No symptoms were observed on the controls. E. sedi is a common Asiatic powdery mildew species infecting many crassulaceous plants (1,2) and was introduced to Eastern Europe from Asia (2). To our knowledge, this is the first report of E. sedi in North America. References: (1) U. Braun. Beih. Nova Hedwigia 89:1, 1987. (2) U. Braun. The Powdery Mildews (Erysiphales) of Europe. Gustav Fisher Verlag, Jena, 1995.