Phytophthora capsici: Recent Progress on Fundamental Biology and Disease Management 100 Years After Its Description.

Phytophthora capsici is a destructive oomycete pathogen of vegetable, ornamental, and tropical crops. First described by L.H. Leonian in 1922 as a pathogen of pepper in New Mexico, USA, P. capsici is now widespread in temperate and tropical countries alike. Phytophthora capsici is notorious for its capability to evade disease management strategies. High genetic diversity allows P. capsici populations to overcome fungicides and host resistance, the formation of oospores results in long-term persistence in soils, zoospore differentiation in the presence of water increases epidemic potential, and a broad host range maximizes economic losses and limits the effectiveness of crop rotation. The severity of disease caused by P. capsici and management challenges have led to numerous research efforts in the past 100 years. Here, we discuss recent findings regarding the biology, genetic diversity, disease management, fungicide resistance, host resistance, genomics, and effector biology of P. capsici.

Disease Resistance to Multiple Fungal and Oomycete Pathogens Evaluated Using a Recombinant Inbred Line Population in Pepper.

Incorporating disease resistance into cultivars is a primary focus of modern breeding programs. Resistance to pathogens is often introgressed from landrace or wild individuals with poor fruit quality into commercial-quality cultivars. Sites of multiple disease resistance (MDR) are regions or "hot spots" of the genome with closely linked genes for resistance to different pathogens that could enable rapid incorporation of resistance. An F2-derived F6 recombinant inbred line population from a cross between 'Criollo de Morelos 334' (CMM334) and 'Early Jalapeno' was evaluated in inoculated fruit studies for susceptibility to oomycete and fungal pathogens: Phytophthora capsici, P. nicotianae, Botrytis cinerea, Fusarium oxysporum, F. solani, Sclerotinia sclerotiorum, Alternaria spp., Rhizopus oryzae, R. stolonifer, and Colletotrichum acutatum. All isolates evaluated were virulent on pepper. Significant differences in disease susceptibility were identified among lines for each of the pathogens evaluated. P. capsici was the most virulent pathogen, while R. oryzae and one Sclerotinia isolate were the least virulent. Quantitative trait loci associated with resistance were identified for Alternaria spp. and S. sclerotiorum. Positive correlations in disease incidence were detected between Alternaria spp. and F. oxysporum, F. solani, and C. acutatum, as well as between C. acutatum and Botrytis spp., F. oxysporum, F. solani, and P. capsici. No sites of MDR were identified for pathogens tested; however, positive correlations in disease incidence were detected among pathogens suggesting there may be genetic linkage among resistance genes in CM334 and Early Jalapeno.

Regional and Temporal Population Structure of Pseudoperonospora cubensis in Michigan and Ontario.

Cucurbit downy mildew (CDM), caused by the oomycete pathogen Pseudoperonospora cubensis, is a devastating disease that affects cucurbit species worldwide. This obligate, wind-dispersed pathogen does not overwinter in Michigan or other northern regions and new isolates can enter the state throughout the growing season. To evaluate the regional and temporal population structure of P. cubensis, sporangia from CDM lesions were collected from cucurbit foliage grown in Michigan and Ontario field locations in 2011. Population structure and genetic diversity were assessed in 257 isolates using nine simple sequence repeat markers. Genetic diversity was high for isolates from Michigan and Canada (0.6627 and 0.6131, respectively). Five genetic clusters were detected and changes in population structure varied by site and sampling date within a growing season. The Michigan and Canada populations were significantly differentiated, and a unique genetic cluster was detected in Michigan.

Resistance to Crown and Root Rot Caused by Phytophthora capsici in a Tomato Advanced Backcross of Solanum habrochaites and Solanum lycopersicum.

Phytophthora capsici causes devastating disease on many vegetable crops, including tomato and other solanaceous species. Solanum habrochaites accession LA407, a wild relative of cultivated tomato, has shown complete resistance to four P. capsici isolates from Michigan cucurbitaceous and solanaceous crops in a previous study. Greenhouse experiments were conducted to evaluate 62 lines of a tomato inbred backcross population between LA407 and the cultivated tomato 'Hunt 100' and 'Peto 95-43' for resistance to two highly virulent P. capsici isolates. Roots of 6-week-old seedlings were inoculated with each of two P. capsici isolates and maintained in the greenhouse. Plants were evaluated for wilting and plant death three times per week for 5 weeks. Significant differences were observed in disease response among the inbred tomato lines. Most lines evaluated were susceptible to P. capsici isolate 12889 but resistant to isolate OP97; 24 tomato lines were resistant to both isolates. Heritability of Phytophthora root rot resistance was high in this population. Polymorphic molecular markers located in genes related to resistance and defense responses were identified and added to a genetic map previously generated for the population. Resistant lines and polymorphic markers identified in this study are a valuable resource for development of tomato varieties resistant to P. capsici.

Susceptibility of Greenhouse Ornamentals to Phytophthora capsici and P. tropicalis.

The susceptibility of fabaceous (Lupinus and Lathyrus spp.) and solanaceous (Calibrachoa, Browallia, Nicotiana, Nierembergia, and Petunia spp.) ornamental plants compared with straightneck squash (Cucurbita pepo) inoculated with Phytophthora capsici and P. tropicalis was investigated in greenhouse studies. Four P. capsici isolates and one P. tropicalis isolate were evaluated. Flowering tobacco (Nicotiana × sanderae), sweet pea (Lathyrus latifolius), lupine (Lupinus polyphyllus), squash, and million bells (Calibrachoa × hybrida) were susceptible to P. capsici and P. tropicalis. Bush violet (Browallia speciosa) and cup flower (Nierembergia scoparia) were not susceptible to either pathogen. Petunia (Petunia × hybrida) was susceptible to P. capsici but not P. tropicalis. Area under the plant growth curve (AUPGC) was also affected in some susceptible plants. AUPGC was significantly different in inoculated plants compared with the untreated controls of Nicotiana and Calibrachoa. In addition, six Calibrachoa cultivars were evaluated for susceptibility to P. capsici and P. tropicalis in a separate experiment. Although all Calibrachoa cultivars were susceptible to P. capsici and P. tropicalis, 'Celebration Purple Star' displayed limited disease following inoculation with either pathogen. Calibrachoa 'Cabaret' and 'Can-Can' inoculated with P. capsici or P. tropicalis displayed significant differences in AUPGC compared with the uninoculated controls. 'Callie', 'Million Bells Cherry Pink', and 'Superbells' had significantly reduced AUPGC compared with the controls when inoculated with P. tropicalis but not P. capsici. These results expand the host range of P. capsici to include Calibrachoa spp., L. polyphyllus, and Lathyrus latifolius, and P. tropicalis to include L. latifolius, Nicotiana spp., and straightneck squash.

Field Response of Cucurbit Hosts to Pseudoperonospora cubensis in Michigan.

Downy mildew, caused by Pseudoperonospora cubensis, is a severe foliar disease of many cucurbit crops worldwide. Forty-one cucurbit cultigens (commercial cultivars and plant introductions) from five genera (Cucumis, Citrullus, Cucurbita, Lagenaria, and Luffa) were assessed for susceptibility to P. cubensis in a research field exposed to natural inoculum in Michigan. Eight cultigens from a differential set for pathotype determination were included within the 41 cultigens to detect changes in dominant P. cubensis pathotypes present. No pathotype differences were found between 2010 and 2011. Cucumis melo cultigen MR-1 was less susceptible to Michigan P. cubensis populations than other C. melo cultigens. No symptoms or signs of infection were detected on cultigens of Cucurbita moschata and C. pepo. Disease onset was later in 2011 than 2010; greater than 90% disease severity in pickling cucumber 'Vlaspik' was observed in both years. This study confirmed that Cucumis is the most susceptible cucurbit genus, while Citrullus and Cucurbita cultigens were the least susceptible genera to populations of P. cubensis in Michigan. Area under the disease progress curve values indicated that disease progress was limited on all Citrullus cultigens compared with Cucumis cultigens, and pathogen sporulation was not detected under field conditions. Future studies should evaluate the ability of a reduced fungicide program to control downy mildew on less susceptible Cucumis melo 'Edisto 47', 'Primo', 'Athena', 'Strike', 'Ananas', 'Banana', and 'Tam-Dew'. Many of the melon cultivars evaluated were selected on the basis of reported resistance to downy mildew, yet they showed significant disease symptoms. It is important to evaluate candidate cultigens for resistance to local P. cubensis populations.

Evaluation of a Diverse, Worldwide Collection of Wild, Cultivated, and Landrace Pepper (Capsicum annuum) for Resistance to Phytophthora Fruit Rot, Genetic Diversity, and Population Structure.

Pepper is the third most important solanaceous crop in the United States and fourth most important worldwide. To identify sources of resistance for commercial breeding, 170 pepper genotypes from five continents and 45 countries were evaluated for Phytophthora fruit rot resistance using two isolates of Phytophthora capsici. Genetic diversity and population structure were assessed on a subset of 157 genotypes using 23 polymorphic simple sequence repeats. Partial resistance and isolate-specific interactions were identified in the population at both 3 and 5 days postinoculation (dpi). Plant introductions (PIs) 640833 and 566811 were the most resistant lines evaluated at 5 dpi to isolates 12889 and OP97, with mean lesion areas less than Criollo de Morelos. Genetic diversity was moderate (0.44) in the population. The program STRUCTURE inferred four genetic clusters with moderate to very great differentiation among clusters. Most lines evaluated were susceptible or moderately susceptible at 5 dpi, and no lines evaluated were completely resistant to Phytophthora fruit rot. Significant population structure was detected when pepper varieties were grouped by predefined categories of disease resistance, continent, and country of origin. Moderately resistant or resistant PIs to both isolates of P. capsici at 5 dpi were in genetic clusters one and two.

QTL mapping of fruit rot resistance to the plant pathogen Phytophthora capsici in a recombinant inbred line Capsicum annuum population.

Phytophthora capsici is an important pepper (Capsicum annuum) pathogen causing fruit and root rot, and foliar blight in field and greenhouse production. Previously, an F6 recombinant inbred line population was evaluated for fruit rot susceptibility. Continuous variation among lines and partial and isolate-specific resistance were found. In this study, Phytophthora fruit rot resistance was mapped in the same F6 population between Criollo del Morelos 334 (CM334), a landrace from Mexico, and 'Early Jalapeno' using a high-density genetic map. Isolate-specific resistance was mapped independently in 63 of the lines evaluated and the two parents. Heritability of the resistance for each isolate at 3 and 5 days postinoculation (dpi) was high (h(2) = 0.63 to 0.68 and 0.74 to 0.83, respectively). Significant additive and epistatic quantitative trait loci (QTL) were identified for resistance to isolates OP97 and 13709 (3 and 5 dpi) and 12889 (3 dpi only). Mapping of fruit traits showed potential linkage with few disease resistance QTL. The partial fruit rot resistance from CM334 suggests that this may not be an ideal source for fruit rot resistance in pepper.

Evaluation of Pepper Fruit for Resistance to Phytophthora capsici in a Recombinant Inbred Line Population, and the Correlation with Fruit Shape.

Phytophthora capsici causes fruit, root, and foliar blight on pepper (Capsicum annuum) in field production. Breeding for disease-resistant commercial pepper cultivars is essential to long-term management of P. capsici. In this study, the severity of Phytophthora fruit rot was evaluated in an F6 recombinant inbred line population between CM334, a landrace from Mexico, and the commercial 'Early Jalapeño'. The two parents and 67 progeny lines were evaluated for fruit rot resistance at 3 and 5 days post inoculation (dpi) using three P. capsici isolates. Fruit shape was also evaluated for each line, and the correlation between shape and disease symptoms was investigated. Significant differences were detected among lines in lesion area measured 3 and 5 dpi, and in phenotypic traits (fruit length, width, and shape index). Of the fruit phenotypic traits measured, only fruit shape index had a significant, albeit weak (r = 0.2892, P = 0.02), correlation with lesion area when inoculated, and with only one of the three isolates of P. capsici evaluated. These results suggest that breeding for fruit rot resistance in pepper will have minimal linkage with fruit shape in the CM334 background.

Evaluation of Eggplant Rootstocks and Pepper Varieties for Potential Resistance to Isolates of Phytophthora capsici from Michigan and New York.

Phytophthora capsici is a soilborne pathogen of major economic importance in pepper, and of less importance in tomato and eggplant production. As soil fumigation becomes more expensive and limited, and fungicide insensitivity of P. capsici isolates becomes more prevalent, grafting is quickly becoming an industry-favored method to control soilborne diseases. Greenhouse experiments were performed to evaluate an eggplant cultivar (Classic), two eggplant lines (EG195, EG203), a pepper line (CM334), and three pepper cultivars (Paladin, Camelot, and Red Knight) for root rot resistance to 14 P. capsici isolates. The isolates showed various degrees of virulence between pepper and eggplant in both experiments. Both eggplant and one pepper lines showed moderate resistance to the most virulent isolates tested in experiment one. The partially resistant pepper cultivar, Paladin, was significantly more susceptible than CM334 and the eggplant lines, but was still resistant to most isolates. In the second experiment, the eggplant cultivar Classic and the susceptible pepper cultivar Red Knight were both susceptible to most isolates tested, while EG203 and EG195 were resistant to most isolates. The two eggplant breeding lines, EG195 and EG203, showed moderate resistance to all isolates tested in both experiments. This is the first reported evaluation of eggplant resistance to P. capsici. Further research is warranted to test eggplant lines EG195 and EG203 for resistance to a wide range of soilborne pests and to evaluate their usefulness as P. capsici-resistant rootstocks for peppers, tomatoes, and eggplants.

First Report of Root and Crown Rot of Wasabi (Wasabia japonica Matsum.) Caused by Phytophthora cryptogea in Michigan.

In September 2011, a Phytophthora sp. was isolated from wasabi (Wasabia japonica Matsum.) grown commercially in hydroponic culture in a large production facility in southwest Michigan. Approximately 20% of the plants were affected, resulting in serious losses for the grower. Plants exhibited severe wilting and root and crown rot, with soft water-soaked lesions on the crown and dark lesions on the roots. Small pieces of root tissue with dark lesions were excised and plated onto potato dextrose agar and unclarified V8 agar plates amended with 25 ppm of benomyl, 100 ppm of ampicillin, 30 ppm of rifampicin, and 100 ppm of pentachloronitrobenzene. Isolates of a Phytophthora sp. were recovered from root tissue. Isolates produced sporangia abundantly in culture. Sporangia averaged 48 µm long × 34 µm wide and were ellipsoid to ovoid, occasionally obpyriform, and were nonpapillate and noncaducous. Distinct hyphal swellings were noted and chlamydospores were observed rarely in culture. The isolate used for inoculations did not produce oospores alone in culture but was able to produce oospores when paired with an A1 culture of P. capsici and incubated in the dark. Oospores were not observed when the isolate was paired with an A2 culture of P. capsici. No growth was observed at 35°C, and the isolate was identified as Phytophthora cryptogea based on morphological and physiological traits. Pathogen identity was further confirmed using PCR primers specific to P. cryptogea (1). In addition, a BLAST search was conducted using the nucleotide database collection in GenBank comparing our isolate against Phytophthora spp., with 99% sequence similarity to P. cryptogea in two sequenced genes, beta tubulin and cytochrome c oxidase 1 (2). Sequences for the isolate were deposited in the GenBank database under accession numbers JX041520 and JX041521. To fulfill Koch's postulates, six small, potted wasabi seedlings were inoculated by placing 3 g of 1-month-old infested millet (100 g of millet, 72 ml of distilled water, 0.08 g of asparagine, and 10 7-mm diameter V8 agar plugs with actively growing P. cryptogea) onto the soil surface of each pot under coconut coir mulch. Plants were watered heavily after soil infestation and as needed thereafter. Three control plants were inoculated with sterile millet seed. The experiment was repeated once. Wilting was observed within 5 and 7 days, respectively, in the first and second experiment. All six inoculated plants were severely wilted within 25 and 56 days, respectively, except for a single plant in the second experiment that never wilted. Root and crown rot was observed on wilted plants and dark lesions could be observed on root tissue. P. cryptogea was recovered from five of the six plants inoculated in each experiment. None of the control plants in either experiment displayed symptoms of wilting, and the pathogen was not recovered from these plants when pieces of root tissue were excised and plated onto amended V8 agar. To our knowledge, this is the first report of P. cryptogea causing crown and root rot of wasabi. References: (1) D. Minerdi et al. Eur. J. Plant Pathol. 122:227, 2008. (2) L. M. Quesada-Ocampo et al. Phytopathology 101:1061, 2011.