Genetic analyses of Xanthomonas axonopodis pv. dieffenbachiae strains reveal distinct phylogenetic groups.

A comprehensive analysis of 175 Xanthomonas axonopodis pv. dieffenbachiae strains isolated from 10 Araceae hosts was done to identify pathogen variation. The strains were subjected to repetitive extragenic palindromic sequence polymerase chain reaction and four major phylogenetic clusters were generated. A subset of 40 strains isolated from Anthurium, Dieffenbachia, and Syngonium was further defined by amplified fragment length polymorphism and fatty acid methyl ester analysis and the same four phylogenetic clusters were observed. Comparison of representative strains in the first three clusters using DNA-DNA hybridization and multilocus sequence analysis supports the previous reclassification of strains in cluster I, including the X. axonopodis pv. dieffenbachiae pathovar reference strain (LMG695), to X. citri. Our research findings indicate that strains in cluster I, isolated primarily from anthurium, probably represent an undescribed pathovar. Other phylogenetic subclusters consisting primarily of strains isolated from xanthosoma and philodendron in clusters III and IV, respectively, may yet represent other undescribed species or pathovars of Xanthomonas.

Outbreak of Cucurbit Powdery Mildew on Watermelon Fruit Caused by Podosphaera xanthii in Southwest Florida.

Cucurbit powdery mildew caused by the obligate parasite Podosphaera xanthii occurs commonly on foliage, petioles, and stems of most cucurbit crops grown in the United States. (3). However, in the field, fruit infection on cucurbits including watermelon (Citrullus lanatus), is rarely, if ever, observed (2). Consequently, it was atypical when severe powdery mildew-like symptoms were observed on seedless and seeded watermelon fruit on several commercial farms in southwestern Florida during November and December 2010. Severe powdery mildew was also observed on 'Tri-X 313' and 'Mickey Lee' fruit grown at SWFREC, Immokalee, FL. Infected fruit developed poorly and were not marketable. Powdery mildew symptoms were mainly observed on young immature fruit, but not on mature older fruit. Abundant powdery mildew conidia occurred on fruit surface, but not on the leaves. Conidia were produced in chains and averaged 35 × 21 µm. Observation of conidia in 3% KOH indicated the presence of fibrosin bodies commonly found in the cucurbit powdery mildew genus Podosphaera (3). Orange-to-dark brown chasmothecia (formerly cleisthothecia) containing a single ascus were detected on the surface of some fruit samples. Conidial DNA was subjected to PCR using specific primers designed to amplify the internal transcribed spacer (ITS) region of Podosphaera (4). The resulting amplicons were sequenced and found to be 100% identical to the ITS sequences of P. xanthii in the NCBI database (D84387, EU367960, AY450961, AB040322, AB040315). Sequences from the watermelon fruit isolate were also identical to several P. fusca (synonym P. xanthii), P. phaseoli (GQ927253), and P. balsaminae (AB462803) sequences. On the basis of morphological characteristics and ITS sequence analysis, the pathogen infecting watermelon fruit can be considered as P. xanthii (1,3,4). The powdery mildew isolate from watermelon fruit was maintained on cotyledons of squash (Cucurbita pepo, 'Early Prolific Straight Neck'). Cotyledons and leaves of five plants each of various cucurbits and beans were inoculated with 10 µl of a conidial suspension (105conidia/ml) in water (0.02% Tween 20). Two weeks after inoculation, abundant conidia were observed on cucumber (Cucumis sativus, 'SMR-58') and melon (Cucumis melo) powdery mildew race differentials 'Iran H' and 'Vedrantais'. However, no growth was observed on melon differentials 'PI 414723', 'Edisto 47', 'PMR 5', 'PMR 45', 'MR 1', and 'WMR 29' (2,3). The powdery mildew isolate from watermelon fruit behaved as melon race 1 (3). Mycelium and conidia were also observed on fruit surface of watermelon 'Sugar Baby' and a susceptible U.S. plant introduction (PI 538888) 3 weeks after inoculation. However, the disease was not as severe as what was observed in the fields in fall 2010. The pathogen did not grow on plants of Impatiens balsamina or on select bean (Phaseolus vulgaris) cultivars ('Red Kidney', 'Kentucky Blue', and 'Derby Bush'), but did grow and produce abundant conidia on 'Pinto bush bean'. Powdery mildew on watermelon fruit in production fields can be considered as a potentially new and serious threat requiring further studies to develop management strategies. References: (1) U. Braun and S. Takamatsu. Schlechtendalia 4:1, 2000. (2) A. R. Davis et al. J. Am. Soc. Hortic. Sci. 132:790, 2007. (3) M. T. McGrath and C. E. Thomas. In: Compendium of Cucurbit Diseases. American Phytopathological Society, St. Paul, MN, 1996. (4) S. Takamatsu and Y. Kano. Mycoscience 42:135, 2001.

Survival and spread of Phytophthora capsici in Coastal Peru.

Phytophthora capsici is a soilborne pathogen that causes significant losses to pepper production in Peru. Our objective was to investigate the mechanisms by which P. capsici is able to survive and spread. During 2005 to 2007, 227 isolates of P. capsici were collected from four species of pepper (Capsicum annum, C. baccatum, C. chinense, and C. pubescens) and tomato (Solanum lycopersicum) at 33 field sites in 13 provinces across coastal Peru. All 227 isolates were of the A2 mating type and amplified fragment length polymorphism (AFLP) analysis indicates that 221 of the isolates had the same genotype. Analyses of six polymorphic single nucleotide polymorphism (SNP) loci showed fixed heterozygosity suggesting a single clonal lineage is widely dispersed. Members of the same clonal lineage were recovered during 2005 to 2007 from geographically separate locations from each of the host types sampled. Our results indicate that clonal reproduction drives the population structure of P. capsici in Peru. The impact of continuous cropping and irrigation from common river sources on the population structure in Barranca Valley are discussed.

Characterization of Phytophthora capsici Causing Foliar and Pod Blight of Snap Bean in Michigan.

Green and yellow snap bean plants with water-soaked foliar lesions, stem necrosis, pod blight, and overall plant decline were observed in four commercial fields in three Michigan counties during 2003 to 2005. All fields were cropped to cucurbits that exhibited symptoms of Phytophthora capsici infection in recent years. In all, 680 isolates of P. capsici were obtained from bean stems, petioles, leaves, and pods; the pathogen was not recovered from roots. Koch's postulates were completed with representative isolates, confirming P. capsici as the causal organism. Select isolates also were pathogenic on cucumber fruit, causing symptoms consistent with P. capsici infection. The majority of the P. capsici isolates collected were sensitive to the fungicide mefenoxam and were of the A1 mating type. Under laboratory conditions, six P. capsici isolates from snap bean (2003) were pathogenic on 12 different commercial bean cultivars, including soybean. Infected bean plants exhibited water-soaked lesions, foliar necrosis, and wilting. We subjected 131 isolates collected from 2003 and 2004 to amplified fragment length polymorphism analysis to investigate diversity among isolates and geographical populations and to determine whether bean P. capsici isolates were similar to isolates from a cucurbit host. This is the first in-depth study of P. capsici on snap bean in Michigan. Although bean cultivars previously were considered a suitable rotation for crops susceptible to P. capsici, this is no longer a recommended practice.

Characterization of Phytophthora capsici from Michigan Surface Irrigation Water.

ABSTRACT Phytophthora capsici infects cucurbitaceous and solanaceous crops worldwide. In free water, P. capsici sporangia release zoospores that may be disseminated by moving surface water. Surface irrigation sources (river system, ponds, and ditches) in three Michigan counties with a history of P. capsici-susceptible crop production were monitored for the pathogen during four growing seasons (2002 to 2005). Pear and cucumber baits were suspended in water at monitoring sites for 3- to 7-day intervals and water temperature was recorded. Baits were washed and lesions were excised and cultured on water agar amended with rifampicin and ampicillin. P. capsici was detected at monitoring sites in multiple years, even when non-host crops were planted nearby. Recovered isolates (N = 270) were screened for sensitivity to the fungicide mefenoxam and characterized for mating type (MT). P. capsici isolates resistant to mefenoxam were common in water sources from southwest and southeast Michigan. Most monitoring sites yielded isolates of a 1:1 ratio of A1:A2 MTs. Amplified fragment length polymorphism analysis of select isolates from 2002 to 2004 indicated a lack of similarity groups persisting over time and in specific geographical locations. Data suggest that P. capsici did not overwinter in any of the surface water sources monitored. Water temperatures were correlated to positive P. capsici detection from all monitoring sites. The frequent detection of P. capsici in surface water used for irrigation in the primary vegetable growing regions in Michigan suggests that this is an important means of pathogen dissemination.