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In 1922, Phytophthora capsici was described by Leon Hatching Leonian as a new pathogen infecting pepper (Capsicum annuum L.), with disease symptoms of root rot, stem and fruit blight, seed rot, and plant wilting and death. Extensive research has been conducted on P. capsici over the last 100 years. This review succinctly describes the salient mile markers of research on P. capsici with current perspectives on the pathogen’s distribution, economic importance, epidemiology, genetics and genomics, fungicide resistance, host susceptibility, pathogenicity mechanisms, and management.

Bacterial spot (BS) of tomato, caused by Xanthomonas gardneri, X. perforans, X. vesicatoria, and X. euvesicatoria, is difficult to control because of the high prevalence of copper- and streptomycin-resistant strains and the lack of resistance cultivars and effective bactericides. The objective of this study was to identify novel growth inhibitors of BS-causing Xanthomonas (BS-X) species by using small molecules (SM; n = 4,182). Several SMs (X1, X2, X5, X9, X12, and X16) completely inhibited the growth of BS-X isolates (n = 68 X. gardneri, 55 X. perforans, 4 X. vesicatoria, and 32 X. euvesicatoria) at ≥12.5 µM by disrupting Xanthomonas cell integrity through weakening of the cell membrane and formation of pores. These SMs were also effective against biofilm-embedded, copper- and streptomycin-resistant Xanthomonas strains while having minimal impact on other plant pathogenic (n = 20) and beneficial bacteria (n = 12). Furthermore, these SMs displayed equivalent antimicrobial activity against BS-X in seeds and X. gardneri in seedlings compared with conventional control methods (copper sulfate and streptomycin) at similar concentrations while having no detectable toxicity to tomato tissues. SMs X2, X5, and X12 reduced X. gardneri, X. perforans, X. vesicatoria, and X. euvesicatoria populations in artificially infested seeds ≤3.4-log CFU/seed 1 day postinfection (dpi) compared with the infested untreated control (P ≤ 0.05). SMs X1, X2, X5, and X12 reduced disease severity ≤72% and engineered bioluminescent X. gardneri populations ≤3.0-log CFU/plant in infected seedlings at 7 dpi compared with the infected untreated control (P ≤ 0.05). Additional studies are needed to increase the applicability of these SMs for BS management in tomato production.

Plant pathogenic bacteria in the genus Erwinia cause economically important diseases, including bacterial wilt of cucurbits caused by Erwinia tracheiphila. Conventional bactericides are insufficient to control this disease. Using high-throughput screening, 464 small molecules (SMs) with either cidal or static activity at 100 µM against a cucumber strain of E. tracheiphila were identified. Among them, 20 SMs (SM1 to SM20), composed of nine distinct chemical moiety structures, were cidal to multiple E. tracheiphila strains at 100 µM. These lead SMs had low toxicity to human cells and honey bees at 100 µM. No phytotoxicity was observed on melon plants at 100 µM, except when SM12 was either mixed with Silwet L-77 and foliar sprayed or when delivered through the roots. Lead SMs did not inhibit the growth of beneficial Pseudomonas and Enterobacter species but inhibited the growth of Bacillus species. Nineteen SMs were cidal to Xanthomonas cucurbitae and showed >50% growth inhibition against Pseudomonas syringae pv. lachrymans. In addition, 19 SMs were cidal or static against Erwinia amylovora in vitro. Five SMs demonstrated potential to suppress E. tracheiphila when foliar sprayed on melon plants at 2× the minimum bactericidal concentration. Thirteen SMs reduced Et load in melon plants when delivered via roots. Temperature and light did not affect the activity of SMs. In vitro cidal activity was observed after 3 to 10 h of exposure to these five SMs. Here, we report 19 SMs that provide chemical scaffolds for future development of bactericides against plant pathogenic bacterial species.

Experiments were conducted to evaluate potential functional and mechanistic differences in the suppression of Sclerotinia sclerotiorum and S. minor and root-knot nematodes in muck soils by anaerobic soil disinfestation (ASD) using different carbon source amendments. Volatile compounds produced during ASD in muck soil amended with molasses, wheat bran, or mustard greens at 20.2 Mg/ha or a 2% ethanol solution significantly reduced the mycelial growth and number of sclerotia produced by both Sclerotinia spp. compared with the anaerobic control. In amended soils, acetic and butyric acids were detected in concentrations that reduced the viability of sclerotia of both pathogens. Higher concentrations of carbon dioxide were observed in ASD-treated soils, regardless of the amendment, than in the nonamended anaerobic control. Only amendment with wheat bran did not increase the production of methane gas during ASD compared with the controls. Meloidogyne hapla survival was completely suppressed in soils treated with ASD regardless of carbon source. Field trials were conducted in Ohio muck soil to assess survival of sclerotia of both Sclerotinia spp. The viability of sclerotia of both Sclerotinia spp. was significantly reduced in soil subjected to ASD amended with wheat bran (20.2 Mg/ha), molasses (10.1 Mg/ha), or wheat bran (20.2 Mg/ha) plus molasses (10.1 Mg/ha) compared with the controls. A consistent negative correlation between soil reduction and viability of sclerotia of both pathogens was observed. Wheat bran and molasses are both widely available amendments that can be used as ASD carbon sources for the management of soilborne pathogens in muck soils.

Recently, in Central Florida tomato production fields, tomato foliage and fruit were observed with symptoms similar to bacterial speck. Fluorescent pseudomonads were consistently isolated and the strains were characterized by standard LOPAT tests, pathogenicity tests, and genetic characterization using 16S ribosomal RNA (rRNA) sequences and multilocus sequence analysis (MLSA) of conserved housekeeping genes. LOPAT test results indicated that the strains were likely Pseudomonas cichorii. These strains were pathogenic on tomato and were also pathogenic on lettuce, the host for the type strain of P. cichorii. Likewise, strains of P. cichorii isolated in Florida since the early 1980s from hosts other than tomato, along with the type strain, were also pathogenic on tomato. Genetic characterization using 16S rRNA and MLSA confirmed that the strains were most closely related to P. cichorii but varied significantly from the type strain. The Florida P. cichorii strains formed a separate phylogenetic group along with P. cichorii strains isolated from tomato in Tanzania. These strains were different from the previously described morphotypes and genomovars of P. cichorii. Our results indicate the presence of a genetically distinct group of multihost pathogenic P. cichorii strains that have been present in Florida since at least the early 1980s.

Tomato bacterial canker, caused by Clavibacter michiganensis subsp. michiganensis, is transmitted by infected or infested seed and mechanically from plant to plant. Wounds occurring during seedling production and crop maintenance facilitate the dissemination of the pathogen. However, the effects of environmental factors on C. michiganensis subsp. michiganensis translocation and growth as an endophyte have not been fully elucidated. A virulent, stable, constitutively bioluminescent C. michiganensis subsp. michiganensis strain BL-Cmm 17 coupled with an in vivo imaging system allowed visualization of the C. michiganensis subsp. michiganensis colonization process in tomato seedlings in real time. The dynamics of bacterial infection in seedlings through wounds were compared under low (45%) and high (83%) relative humidity. Bacteria multiplied rapidly in cotyledon petioles remaining after clip inoculation and moved in the stem toward both root and shoot. Luminescent signals were also observed in tomato seedling roots over time, and root development was reduced in inoculated plants maintained under both humidity regimes. Wilting was more severe in seedlings under high-humidity regimes. A strong positive correlation between light intensity and bacterial population in planta suggests that bioluminescent C. michiganensis subsp. michiganensis strains will be useful in evaluating the efficacy of bactericides and host resistance.

Lewis Ivey, M. L, McSpadden Gardner, B. B., Opina, N., and Miller, S. A. 2007. Diversity and geographic distribution of Ralstonia solanacearum from eggplant in the Philippines. Phytopathology 97:1467-1475. The diversity of Ralstonia solanacearum strains isolated from eggplant (Solanum melongena) grown in five provinces of the Philippine island group of Luzon was assessed using a recently described hierarchical system. All strains keyed to race 1, biovar 3 or 4. Phylotype-specific multiplex polymerase chain reaction (PCR) indicated that, like most other strains of Asian origin, all the strains in our Philippine collection belong to phylotype I. Taxometric and phylogenetic analyses of partial endoglucanase gene sequences of strains from this collection and those previously deposited into GenBank revealed at least four subgroups among the otherwise monophyletic phylotype I strains. Nucleotide polymorphisms within each subgroup were infrequent and, among the subgroups identified in this study, variation was always

Yang, W., Sacks, E. J., Lewis Ivey, M. L., Miller, S. A., and Francis, D. M. 2005. Resistance in Lycopersicon esculentum intraspecific crosses to race T1 strains of Xanthomonas campestris pv. vesicatoria causing bacterial spot of tomato. Phytopathology 95:519-527. We used molecular markers to identify quantitative trait loci (QTL) that confer resistance in the field to Xanthomonas campestris pv. vesicatoria race T1, a causal agent of bacterial spot of tomato. An F2 population derived from a cross between Hawaii 7998 (H 7998) and an elite breeding line, Ohio 88119, was used for the initial identification of an association between molecular markers and resistance as measured by bacterial populations in individual plants in the greenhouse. Polymorphism in this cross between a Lycopersicon esculentum donor of resistance and an elite L. esculentum parent was limited. The targeted use of a core set of 148 polymerase chain reaction-based markers that were identified as polymorphic in L. esculentum × L. esculentum crosses resulted in the identification of 37 markers that were polymorphic for the cross of interest. Previous studies using an H 7998 × L. pennellii wide cross implicated three loci, Rx1, Rx2, and Rx3, in the hypersensitive response to T1 strains. Markers that we identified were linked to the Rx1 and Rx3 loci, but no markers were identified in the region of chromosome 1 where Rx2 is located. Single marker-trait analysis suggested that chromosome 5, near the Rx3 locus, contributed to reduced bacterial populations in lines carrying the locus from H 7998. The locus on chromosome 5 explained 25% of the phenotypic variation in bacterial populations developing in infected plants. An advanced backcross population and subsequent inbred backcross lines developed using Ohio 88119 as a recurrent parent were used to confirm QTL associations detected in the F2 population. Markers on chromosome 5 explained 41% of the phenotypic variation for resistance in replicated field trials. In contrast, the Rx1 locus on chromosome 1 did not play a role in resistance to X. campestris pv. vesicatoria race T1 strains as measured by bacterial populations in the greenhouse or symptoms in the field. A locus from H 7998 on chromosome 4 was associated with susceptibility to disease and explained 11% of the total phenotypic variation. Additional variation in resistance was explained by plant maturity (6%), with early maturing families expressing lower levels of resistance, and plant habit (6%), with indeterminate plants displaying more resistance. The markers linked to Rx3 will be useful in selection for resistance in elite × elite crosses.

Zhang, J., Hogenhout, S. A., Nault, L. R., Hoy, C. W., and Miller, S. A. 2004. Molecular and symptom analyses of phytoplasma strains from lettuce reveal a diverse population. Phytopathology 94:842-849. Epidemics of aster yellows in lettuce in Ohio are caused by at least seven distinct phytoplasma strains in the aster yellows (AY) group. Five of the strains are newly reported: AY-BW, AY-WB, AY-BD3, AY-SS, and AY-SG. All seven strains were characterized based on symptoms in aster and lettuce, and by polymerase chain reaction (PCR). Strain AY-BD2 (formerly ‘Bolt’) causes yellowing and leaf distortion in lettuce and bolting in aster, whereas strain AY-S (formerly ‘Severe’) causes stunting, leaf clustering, and phyllody. Strain AY-WB causes yellowing and wilting in lettuce and witches’-broom in aster. Strain AY-SG induces horizontal growth in lettuce and aster plants. Strain AY-BW causes chlorosis of emerging leaves and abnormally upright growth of leaf petioles. AY-SS causes symptoms similar to those caused by AY-S but has a different PCR-restriction fragment length polymorphism (RFLP) banding pattern. Strains AY-BD2 and AY-BD-3 cause mild leaf and stem distortion in lettuce but are differentiated by PCR-RFLP. All phytoplasma strains collected from lettuce in Ohio belong to the 16SrI group. AY-WB belongs to the 16SrI-A subgroup and the other six belong to the 16SrI-B subgroup. Five of the seven strains were distinguished from each other by primer typing. The results of phylogenetic analyses of sequences of the 16S rRNA genes were basically consistent with the classification based on PCR-RFLP, in which AY-WB clustered with phytoplasmas of the 16rIA subgroup and the other Ohio lettuce strains clustered with phytoplasmas in the 16SrI-B subgroup. Phytoplasmas are wall-less prokaryotes in the class Mollicutes that infect hundreds of plant species worldwide (25) and cause an array of host symptoms such as vein clearing, yellowing, stunting, and phyllody (20). They are obligate parasites transmitted by phloem-feeding leafhoppers. Aster yellows phytoplasmas (AYP) compose the largest of the phytoplasma groups classified to date (24). Epidemics of aster yellows in vegetable production areas in Ohio and other states in the north-central region of the United States are tied to the immigration of AYP-inoculative aster leafhoppers (Macrosteles quadrilineatus Forbes) in late spring and early summer from states to the south and west (4,6,13). Locally produced leafhoppers acquire and spread AYP within and among host vegetable crops during the growing season. Aster yellows is a potentially devastating disease in lettuce (Lactuca sativa L.), with incidence varying from less than 1 to 100% from year to year in Ohio. A simulation model of aster yellows epidemics in Ohio demonstrated that the pattern of long range dispersal of inoculative leafhoppers, arrangement of host and nonhost crops in an area, and aster yellows latent period in host crops influences the severity of epidemics and yield loss (37). The structure of the model includes rates of AYP acquisition and transmission, as well as leafhopper development rates and latent periods in the plant and insect hosts. The presence of multiple AYP strains in the field could be responsible for the variation in symptoms observed. In addition, interactions among AYP strains in host plants and insect vectors may influence these variables. AYP strains spread in host plants at different rates (17), which may influence acquisition efficiency by leafhoppers feeding on the plants and, subsequently, strain prevalence. In addition, leaf

Diversity in host range, pathogenicity, phenotypic characteristics, repetitive extragenic palindromic polymerase chain reaction (rep-PCR) profiles, and sequence of the 16S-23S rDNA spacer region was examined among 44 Xanthomonas strains isolated from lettuce. Forty-two of the strains were divided into two groups, designated A and B. Seventy percent were Group A, and most of the remaining strains including a reference strain (LMG 938) were Group B. Group A strains induced both local and systemic symptoms, whereas Group B strains caused only distinct necrotic spots. Two strains, including the X. campestris pv. vitians type strain, were distinct from the Group A and B strains and were not pathogenic on lettuce. Analysis of fatty acid profiles, serotype, carbon substrate utilization patterns, and protein fingerprints confirmed this grouping. The Group A and B strains also formed two unique clusters (I and II) by rep-PCR profiling that corresponded to the two groups. Direct sequencing of a PCR-amplified DNA fragment (680 bp) from the 16S-23S rDNA spacer region of four representative strains, however, did not differentiate these groups. Serology and rep-PCR fingerprinting can be used to diagnose and identify X. campestris pv. vitians strains, while the other analyses evaluated are useful for strain characterization.

Populations of Phytophthora spp. were determined by enzyme-linked immunosorbent assay (ELISA) in field soils used for pepper and soybean production in Ohio. Soybean fields were sampled extensively (64 fields, n = 6 samples per field over 2 years) and intensively (4 fields, n = 64 samples per field in 1 year) to assess heterogeneity of P. sojae populations. Four pepper fields (n = 64), three of which had a history of Phytophthora blight (caused by P. capsici), also were sampled intensively during a 6-month period. Mean (m), variance (v), and measures of aggregation (e.g., variance-to-mean ratio [v/m]) of immunoassay values, translated to Phytophthora antigen units (PAU), were related to the disease history in each of the pepper and soybean fields. Mean PAU values for fields in which Phytophthora root rot (soybean) or blight (pepper) had been moderate to severe were higher than in fields in which disease incidence had been low or not observed. A detection threshold value of 11.3 PAU was calculated with values for 64 samples from one pepper field, all of which tested negative for Phytophthora by bioassay and ELISA. Seven of the eight intensively sampled fields contained at least some detectable Phytophthora propagules, with the percentage of positive samples ranging from 1.6 to 73.4. Mean PAU values ranged from 1 to 84 (extensive soybean field sampling), 6 to 24 (intensive soybean field sampling), and 4 to 30 (intensive pepper field sampling); however, variances ranged from 0 to 7,774 (extensive sampling), 30 to 848 (intensive soybean field sampling), and 5 to 2,401 (intensive pepper field sampling). Heterogeneity of PAU was high in most individual soybean and pepper fields, with values of v/m greater than 1, and log(v) increasing with log(m), with a slope of about 2.0. Spatial autocorrelation coefficients were not significant, indicating there was no relationship of PAU values in neighboring sampling units (i.e., field locations) of the intensively sampled fields. Combined results for autocorrelations and v/m values indicate that Phytophthora was highly aggregated in these fields but that the scale of the aggregation (e.g., average focus size) was less than the size of the sampling units. Because of the observed variability, we calculated that sample sizes of 20 or more would be needed to estimate precisely the mean density of Phytophthora in most cases.