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Cercospora leaf spot, caused by Cercospora beticola, is a highly destructive disease of Beta vulgaris subsp. vulgaris worldwide. C. beticola populations are usually characterized by high genetic diversity, but little is known of the relationships among populations from different production regions around the world. This information would be informative of population origin and potential pathways for pathogen movement. For the current study, the genetic diversity, differentiation, and relationships among 948 C. beticola isolates in 28 populations across eight geographic regions were investigated using 12 microsatellite markers. Genotypic diversity, as measured by Simpson’s complement index, ranged from 0.18 to 1.00, while pairwise index of differentiation values ranged from 0.02 to 0.42, with the greatest differentiation detected between two New York populations. In these populations, evidence for recent expansion was detected. Assessment of population structure identified two major clusters: the first associated with New York, and the second with Canada, Chile, Eurasia, Hawaii, Michigan, North Dakota, and one population from New York. Inferences of gene flow among these regions suggested that the source for one cluster likely is Eurasia, whereas the source for the other cluster is not known. These results suggest a shared origin of C. beticola populations across regions, except for part of New York, where population divergence has occurred. These findings support the hypothesis that dispersal of C. beticola occurs over long distances.

Banana bunchy top virus (BBTV), banana streak viruses (BSVs) and cucumber mosaic virus (CMV) are frequently reported infecting bananas globally. Effective control of their spread depends on robust detection of these viruses in propagation stock, planting material, infected nursery plants, and through strict quarantine. We developed single reverse transcription loop-mediated isothermal amplification (RT-LAMP) assays for BBTV, banana streak OL virus (BSV-OL) and CMV that were sensitive, specific, efficient, and completed in less than 60 min. RNA-based RT-LAMP minimized false positives that arose from banana genomes harboring endogenous viral genomes, such as BSVs. RT-LAMP was also more sensitive than RT-PCR in detecting the DNA viruses, BBTV and BSV-OL, in infected plants. We also developed a multiplex assay using three sets of primers specific for each virus to simultaneously detect BBTV, BSV-OL and CMV in a sample of RNA from the same plant. The reliability and convenience of this assay makes it useful for plant quarantine and indexing plants for propagation.

Two diagrammatic ordinal scales are available in the Estimate app (2017 version) for Cercospora leaf spot (CLS) severity on table beet: 10% linear (linear-based diagrammatic scale [LIN]) and logarithmic based (Horsfall–Barratt [HB]). These allow for estimating severity data of four types depending on the system used. A group of 30 raters assigned percentage severity on 30 photographs of diseased table beet leaves during five rounds first without an aid and then using each of the four rating systems in Estimate. In two, the perceived ordinal score of the HB or LIN scale was assigned where severity of the subject fit best. HB2 and LIN2 involved a second choice of unitary severity within the perceived score interval. There was large variation in unaided ability of raters to estimate severity: 13% were accurate (Lin’s concordance correlation [LCC] > 0.9), 23% were inaccurate (LCC < 0.7), and the remaining had moderate accuracy. Larger disparities between assigned and actual ordinal scores (mostly overestimates) occurred using the LIN compared with the HB. The LIN2 produced the most accurate estimates (Lin’s concordance correlation coefficient, ρc = 0.96; generalized bias parameter, Cb = 0.99; Pearson’s correlation coefficient r = 0.95) and the greatest interrater reliability (overall concordance correlation coefficient and intraclass correlation coefficient > 0.93). The two-step process using the 10% linear scale is recommended for severity estimates of CLS in table beet.

In February 2013, an ornamental waxflower (Hoya calycina Schlecter) with leaves displaying concentric chlorotic and necrotic rings surrounding sunken, necrotic lesions typical of tospovirus infection was observed at a community garden in Honolulu, HI. Symptomatic leaf tissue tested negative for Tomato spotted wilt virus (TSWV), a common tospovirus in Hawaii, using a TSWV ImmunoStrips (AgDia, Elkhart, IN) assay following the manufacturer's instructions. Double-stranded RNAs were isolated from a symptomatic leaf and reverse transcribed using random primers (2). The cDNA was then used as template in a universal tospovirus PCR assay using primers gL3637 and gL4435c, which amplify sequences of the L segment encoding the RNA-dependent RNA polymerase of tospoviruses (1). An ~800-bp product was amplified and cloned using pGEM-T Easy (Promega, Madison, WI). Three clones were selected and found to be identical by dye-terminator sequencing performed at the University of Hawaii's Advanced Studies in Genomics, Proteomics, and Bioinformatics laboratory. Following primer sequence trimming, the 773-bp sequence (GenBank Accession No. KF030938) was found to be 97, 88, and 87% identical to Capsicum chlorosis virus (CaCV; a tentative species in the family Bunyaviridae, genus Tospovirus) strains Ch-Har (GU199334), TwTom1 (HM021140), and AIT (DQ256124), respectively. To confirm the presence of CaCV, the cDNA was also used as template in a universal tospovirus PCR assay with primers 3′T12 and TsMCR2 which amplify a region of the S segment of tospoviruses (3). The amplification product from this assay was cloned and sequenced as described above and found to be 93 to 98% identical to CaCV nucleotide sequences present in GenBank. Attempts to detect the CaCV strain in waxflower using a watermelon silver mottle virus and groundnut bud necrosis virus triple antibody sandwich ELISA (AgDia) were unsuccessful. No other plants in the community garden had typical tospovirus-like symptoms; however, samples from tomato (Solanum lycopersicum L.; two samples), chili pepper (Capsicum spp.; four samples), eggplant (Solanum melongena L.; one sample), and passionfruit (Passiflora edulis Sims; one sample) with virus-like symptoms were collected from the garden and had RNA isolated using a NucleoSpin RNA II kit (Macherey-Nagel, Bethlehem, PA). No tospoviruses were detected in any of these samples with the RT-PCR assay using primers gL3637 and gL4435. The waxflower plant infected with CaCV was immediately removed by community garden members and destroyed, preventing any additional serological or biological assays to be performed. CaCV is transmitted by several species of thrips, including Thrips palmi, which is present in Hawaii. Waxflower is not native to Hawaii and it is unclear whether CaCV entered Hawaii in this plant or whether it was infected by viruliferous thrips. A survey for CaCV in known hosts is essential to determine the geographic distribution of CaCV in Hawaii, as this virus poses a considerable threat to tomato, chili pepper, and phalaenopsis orchid production in Hawaii and the United States. References: (1) F.-H. Chu et al. Phytopathology 91:361, 2001. (2) M. J. Melzer et al. Virus Genes 40:111, 2010. (3) M. Okuda and K. Hanada. J. Virol. Methods 96:149, 2001.

An interactive, iterative smartphone application was used on color images to distinguish diseased from healthy plant tissues and calculate percentage of disease severity. The user touches the application’s display screen to select up to eight different colors that represent healthy tissues. The user then moves a threshold slider until only the symptomatic tissues have been transformed into a blue hue. The pixelated image is then analyzed to calculate the disease percentage. This study reports the accuracy, precision, and robustness of Leaf Doctor using six different diseases with typical lesions of varying severity. Estimates of disease severity from Leaf Doctor were highly accurate (R2 ≥ 0.79; Cb ≥ 0.959) compared with estimates obtained from the discipline-standard, Assess. Precision was operationally defined as the ability of a rater to use Leaf Doctor and repeatedly obtain similar percentages of disease severity for the same image. Coefficients of variation were low (0.51 to 14.1%) across all disease datasets but a significant negative relationship was found between the coefficient of variation of estimates and mean disease severity. Other advantages of Leaf Doctor included comparatively less time for image processing, low cost, ease of use, ability to send results by e-mail, and the ability to create realistic standard area diagrams. Leaf Doctor is compatible with iPhone, iPad, and iPod touch and is optimized for iPhone 5. It is available as a free download at the iTunes Store.

The New York table beet industry is expanding and has unique challenges to minimize crop loss in both conventional and organic production. Diseases may reduce plant population density and increase heterogeneity in a stand, reduce the duration of time foliage is healthy, and decrease the yield of marketable roots. Rhizoctonia solani Kuhn and Pythiumultimum Trow are dominant in the pathogen complex affecting crop stand and root health. Cercospora leaf spot (CLS) caused by the fungus, Cercospora beticola Sacc., is a highly destructive disease affecting foliar health. In conventional table beet production, fungicides are applied in-furrow and at emergence for early season and root disease control, and applied to foliage periodically thereafter for foliar disease control. Resistance within C. beticola populations to single-site mode-of-action fungicides poses the most significant threat to the resilience of conventional disease management. An integrated approach to reduce pesticide application when not economically warranted (i.e., a false positive) is urgently required. For foliar disease, improved scheduling of fungicides may reduce usage without loss of disease control. For soilborne diseases, pre-plant quantification of soilborne inoculum may support the selection of fields with lower inoculum densities to minimize risk of early season and root disease. For organic production, some approved products have moderate efficacy for foliar disease control, but strategies to reduce inoculum and select fields at lowest risk of disease will be paramount. Crop rotation has shown promise for disease management, but broad host range of several of the major soilborne pathogens limits the utility of this method in the production region. Enhanced knowledge of cultivar susceptibility to local populations of fungal pathogens responsible for foliar and root diseases is paramount, and adoption of commercially acceptable cultivars with improved resistance to CLS and Rhizoctonia crown and root rot has potential to transform disease management strategies for the New York table beet industry.

Spatial analysis of epiphytotics is essential to develop and test hypotheses about pathogen ecology, disease dynamics, and to optimize plant disease management strategies. Data collection for spatial analysis requires substantial investment in time to depict patterns in various frames and hierarchies. We developed a new approach for spatial analysis of pixelated data in digital imagery and incorporated the method in a stand-alone desktop application called Cluster. The user isolates target entities (clusters) by designating up to 24 pixel colors as nontargets and moves a threshold slider to visualize the targets. The app calculates the percent area occupied by targeted pixels, identifies the centroids of targeted clusters, and computes the relative compass angle of orientation for each cluster. Users can deselect anomalous clusters manually and/or automatically by specifying a size threshold value to exclude smaller targets from the analysis. Up to 1,000 stochastic simulations randomly place the centroids of each cluster in ranked order of size (largest to smallest) within each matrix while preserving their calculated angles of orientation for the long axes. A two-tailed probability t test compares the mean inter-cluster distances for the observed versus the values derived from randomly simulated maps. This is the basis for statistical testing of the null hypothesis that the clusters are randomly distributed within the frame of interest. These frames can assume any shape, from natural (e.g., leaf) to arbitrary (e.g., a rectangular or polygonal field). Cluster summarizes normalized attributes of clusters, including pixel number, axis length, axis width, compass orientation, and the length/width ratio, available to the user as a downloadable spreadsheet. Each simulated map may be saved as an image and inspected. Provided examples demonstrate the utility of Cluster to analyze patterns at various spatial scales in plant pathology and ecology and highlight the limitations, trade-offs, and considerations for the sensitivities of variables and the biological interpretations of results. The Cluster app is available as a free download for Apple computers at iTunes, with a link to a user guide website.

Morphological and molecular studies confirmed Paracercospora egenula as the primary cercosporoid fungus causing leaf spot of eggplant (Solanum melongena) in Hawaii. This is the first confirmed record of P. egenula in Hawaii, although it was previously reported as C. solani-melongenae, based on an interception at Oahu, Hawaii, in 1946. Another cercosporoid species, Cercospora melongenae, reported to cause leaf spot on eggplant in Hawaii, was not detected.

Taro leaf blight (TLB), caused by the oomycete pathogen Phytophthora colocasiae, is a worldwide disease that threatens the sustainable cultivation of the tropical root crop taro (Colocasia esculenta). To evaluate taro germplasm from Asia, Hawai‘i, and several South Pacific Islands for resistance to TLB, 119 cultivars were planted along the Hamakua Coast of Hawai‘i (mean annual rainfall of 130 inches) in plots containing five or 10 plants that were replicated over time from 1993 through 2005. Fresh and dry weights of corms were measured after about nine months, with rotten portions removed and weighed. When epidemics of TLB occurred (in nine out of 12 years), visual estimates of disease severity on leaves were assessed using a modified Horsfall–Barratt scale. The correlations between mean dry weight yields for each cultivar and mean severity of TLB, and, respectively, between mean yields and mean severity of corm rots were calculated. As severity of TLB or severity of corm rots increased (suggesting increased susceptibility of particular cultivars to TLB or corm rots), mean dry weight yields decreased significantly (r2 = 0.37 and 0.22, respectively). “Multiple comparisons with the best” (MCB) were conducted on fresh and dry weight yields, severity of TLB, severity of corm rots, percentage dry matter of corm, and consumer acceptance. Five cultivars were found to be “among the best” with: 1) fresh or dry weight yields that did not differ from the highest level; 2) severity ratings for TLB that were significantly lower than the highest level, suggesting TLB resistance; and 3) percentage of corm rots that were lower than the highest level, suggesting disease resistance. These cultivars, four of which originated from Palau, were Dirratengadik, Merii, Ngesuas, Ochelochel, and Sawa Bastora. Two commercial cultivars from Hawai‘i, Bun Long and Maui Lehua, had fresh and dry weight yields that were significantly lower than the maximum and severity of TLB injury that did not differ from the highest level, indicating that conventional breeding of taro to improve TLB resistance could improve yields of commercial taro cultivars, particularly in areas where epidemics of TLB occur.

Mechanisms of resistance to rice stripe disease in a Chinese rice cultivar (Oryza sativa L., cv. Zhendao 88) were determined, and molecular markers for the resistance gene were identified. Single tillers at the seedling stage were inoculated with Rice stripe virus (RSV) and its vector, the small brown planthopper (SBPH) Laodelphax striatellus Fallen, to test for non-preference and antibiosis. The inheritance of resistance in the F2 and F2 : 3 lines from the cross cvs Zhendao 88× Wuyujing No. 3 was also examined by single-tiller inoculation. Cv. Zhendao 88 was highly resistant to RSV and weakly resistant to SBPH. The resistance gene was mapped by SSR and RAPD analyses to rice chromosome 11 within 4.7 cm of a SSR marker RM229 and a RAPD marker OPO11. Data and inheritance analysis indicated that rice stripe disease resistance in cv. Zhendao 88 was derived principally from resistance to RSV and controlled by a single dominant gene. Breeding for rice stripe resistance could be accelerated by using cv. Zhendao 88 as a resistant parent if the linked marker for virus resistance were used in a marker-assisted progeny selection programme.

Genotyping-by-sequencing (GBS) was conducted on 333 Cercospora isolates collected from Beta vulgaris (sugar beet, table beet and swiss chard) in the USA and Europe. Cercospora beticola was confirmed as the species predominantly isolated from leaves with Cercospora leaf spot (CLS) symptoms. However, C. cf. flagellaris also was detected at a frequency of 3% in two table beet fields in New York. Resolution of the spatial structure and identification of clonal lineages in C. beticola populations using genome-wide single nucleotide polymorphisms (SNPs) obtained from GBS was compared to genotyping using microsatellites. Varying distance thresholds (bitwise distance = 0, 1.854599 × 10-4, and 1.298 × 10-3) were used for delineation of clonal lineages in C. beticola populations. Results supported previous reports of long distance dispersal of C. beticola through genotype flow. The GBS-SNP data set provided higher resolution in discriminating clonal lineages; however, genotype identification was impacted by filtering parameters and the distance threshold at which the multi-locus genotypes (MLGs) were contracted to multi-locus lineages. The type of marker or different filtering strategies did not impact estimates of population differentiation and structure. Results emphasize the importance of robust filtering strategies and designation of distance thresholds for delineating clonal lineages in population genomics analyses that depend on individual assignment and identification of clonal lineages. Detection of recurrent clonal lineages shared between the USA and Europe, even in the relaxed-filtered SNP data set and with a conservative distance threshold for contraction of MLGs, provided strong evidence for global genotype flow in C. beticola populations. The implications of intercontinental migration in C. beticola populations for CLS management are discussed.

The survival of the bacterial pathogen, Xanthomonas campestris pv. campestris (Xcc), was studied in plant debris-infested soil with different matric potentials (0, −10, −30, −50, −100, −200, and −900 kPa), and on the phylloplane of crops used for rotation with cabbage. Populations of cellulolytic and proteolytic microorganisms were studied in relation to soil matric potential and Xcc. The survival of Xcc was negatively correlated (r=−0.710; P=0.06) with soil matric potential and with the abundance of cellulolytic microorganisms (P=0.05). In saturated soil, Xcc survived for only 19–28 days, while at −900 kPa there was no significant change in the bacterial population within this period. Survival of Xcc on the phylloplane of cabbage (Brassica oleracea var. capitata) (host) was similar to that on mustard and lettuce (48 days), whereas the pathogen was detected for only 9 days on rice. It appears that high matric potentials reduced populations of Xcc in soil, whereas epiphytic survival of this pathogen depended on the plant species.

The infection process of bacterial blight of anthurium was monitored with a bioluminescent strain of Xanthomonas campestris pv. dieffenbachiae. The relationship between symptom expression on infected leaves (assessed visually) and the extent of bacterial movement within tissues (evaluated by bioluminescence emission) varied among anthurium cultivars. In several cultivars previously considered susceptible on the basis of symptom development alone, bacterial invasion of leaves extended far beyond the visually affected areas. In other cultivars previously considered resistant, bacterial invasion was restricted to areas with visible symptoms. In three cultivars previously considered resistant, leaves were extensively invaded by the bacterium, and yet few or no symptoms were seen on infected leaves. The pathogen was consistently recovered from leaf sections emitting bioluminescence but not from sections emitting no light. At an early stage of infection, no significant differences in the percentages of infected areas as determined by visual assessment were observed in any of the cultivars. However, differences among cultivars were detected by bioluminescence as the disease progressed, because bacterial invasion was not always accompanied by symptom expression. In susceptible cultivars, the advancing border of infection was 5 to 10 cm inward from the margins of the visible symptoms and often reached to the leaf petiole even when symptoms were visible in

A homothallic, papillate Phytophthora species causing foliar and fruit blight of noni (Morinda citrifolia var. citrifolia) in Hawaii was identified. The asexual phase of this species is characterized by the production of umbellate sporangiophores and papillate sporangia that are ellipsoid and obpyriform with conspicuously tapered bases and possess caducous, medium to long pedicels. The sexual phase is characterized by the production of oogonia with tapered bases, small amphigynous antheridia and thick-walled, plerotic oospores. The morphology of the taxon does not match any of the valid 95 Phytophthora species described to date. Phylogenetic analysis based on sequences of the internal transcribed spacer rDNA region (ITS) and the translation elongation factor 1 alpha (EF-1 alpha) of this taxon and those from other Phytophthora species from GenBank and the Phytophthora database indicates that the new taxon is most closely related to species in ITS clade 10, including P. kernoviae, P. boehmeriae and the recently described P. gallica. The most closely related species is P. kernoviae, an invasive plant pathogen causing bleeding stem lesions on forest trees (beech, Fagus sylvatica) and foliar necrosis of ornamentals (rhododendron, pieris and magnolia) in the UK, and isolated in New Zealand from necrotic cherimoya shoots and fruits and soil. Although the morphological characters of the sexual phase of P. morindae and P. kernoviae are similar, the umbellate sporangiophores produced by the new taxon marks the main morphological distinction. In this paper we describe the morphological characteristics, the phylogenetic relationships and pathogenicity characteristics that support the description of this taxon as a new species with the proposed name Phytophthora morindae sp. nov.

Spatiotemporal distance class analysis is proposed as a new form of spatiotemporal analysis of intensively mapped, binary data. The method detects and quantifies attributes of nonrandom patterns of disease increase in regularly spaced plant populations. The method tests the hypothesis that healthy plants in a population have an equal (random) chance of becoming diseased in the period between two disease assessment dates. Mapped disease incidence evaluations from two assessment dates are needed for the analysis. Expected spatial patterns of diseased plants for the second assessment date are generated by assigning the number of newly diseased plants to random spatial positions among the healthy plant population observed on the first assessment date. Distance class analysis techniques are used to compare these expected patterns with the actual spatial pattern observed on the second assessment date. The number, location, and configuration of significant distance classes are used to evaluate the randomness of the observed spatial distribution. Hypothetical examples and data from an epidemic of citrus variegated chlorosis in Brazil and an epidemic of papaya ringspot in Hawaii are presented to illustrate the procedure

Disease progress, host growth and defoliation, and spatial patterns for a multiple pathogen, leaf spot disease complex on white clover (Trifolium repens) were monitored in four 6-wk growth periods during 1990-1991 in a natural, 10-ha pasture of white clover and tall fescue (Festura arundinacea) in North Carolina. The disease complex comprised summer blight (Rhizoctonia solani), black spot (Pseudomonas andropogonis), Stagonospora leaf spot (Stagonospora meliloti), Cercospora leaf spot (Cercospora zebrina), Curvularia leaf spot (Curvularia trifolii), anthracnose (Colletotrichum trifolii), sooty blotch (Polythrincium trifolii), and rust (Uromyces sp.) []

Incidence and spatial patterns of eight foliar pathogens and diseases were monitored during two 6-wk growth periods per year from June to September in 1990 and 1991 on regularly spaced plants of white clover (Trifolium repens) planted in a tall fescue (Festuca arundinacea) sward in a 10-ha pasture in Wake County, North Carolina. Disease ratings were made in four plots on a total of 512 plants in eight square lattices of 64 plants each. Each plot comprised two proximal lattices of either the virus-susceptible white clover cultivar, Regal, or the Southern Regional Virus Resistant white clover germ plasm []

Incidence of pathogen species and their patterns of association in leaf spot disease complex on white clover (Trifolium repens) were observed in plots and adjacent areas in a pasture during 1990-1991 and in a monthly (July-October) survey of commercial pastures during 1991. Leaf samples from seven pastures in five counties (Chatham, Franklin, Nash, Orange, and Wake) in the Piedmont region of North Carolina and one county in western North Carolina (Yancey) were examined (...)

An isolate of Cercospora zebrina from white clover (Trifolium repens) was pathogenic to 24 species in 12 genera of legumes in greenhouse tests. This report is apparently the first evidence of pathogenicity of a white clover isolate of Cercospora to species of Apios, Arachis, Glycine, Lespedeza, Lotus, Phaseolus, and Vigna. The isolate was not always more virulent on white clover; pea cultivars (Pisum sativum) were more severely diseased. Five isolates of C. zebrina from white clover in four counties in North Carolina manifested significant (P