Effect of oxathiapiprolin on asexual life stages of Phytophthora capsici and disease development on vegetables

Phytophthora blight caused by Phytophthora capsici is a serious disease in the production of peppers and other vegetables worldwide. Application of fungicides is an important component in developing effective disease management programmes. However, resistance in P. capsici populations to some commonly used fungicides has been documented. Identification of effective new fungicides with different mode of actions is highly desirable. This study was conducted to determine baseline sensitivity of P. capsici isolates to oxathiapiprolin, the first member of a new class of isoxazoline fungicides, and efficacy of this compound for reduction of Phytophthora blight on bell pepper. A collection of 126 P. capsici isolates were evaluated and all the isolates were sensitive to oxathiapiprolin. EC₅₀ values of oxathiapiprolin in inhibiting mycelial growth, sporangium formation and zoospore germination of 25 selected isolates averaged 0.001, 0.0003 and 0.54 µg mL^?1, respectively. It appeared that asexual life stages of P. capsici were more sensitive to oxathiapiprolin than other compounds used for control of oomycete pathogens. In field studies, oxathiapiprolin applied at different rates through drip irrigation tubes, or by soil drench plus foliar sprays, reduced Phytophthora blight and increased pepper yield significantly. This is the first report of the efficacy of oxathiapiprolin in suppression of P. capsici, which indicates that oxathiapiprolin is effective in inhibiting the pathogen and has the promise to be a viable option for managing Phytophthora blight in bell pepper production.     

Susceptibility assessment of bell pepper genotypes to crown and root rot disease

Bell Pepper (Capsicum annuum) is one of the important vegetable crops with valuable food sources, which is used almost around the world. Crown and root rot disease caused by Phytophthora capsici is one of the most important diseases of bell pepper in Iran. The present study was conducted to evaluate the susceptibility of different varieties of bell pepper to crown and root rot disease under glasshouse condition. Fourteen commonly planted genotypes of bell pepper in Iran were evaluated for their susceptibility to infection with the pathogen. For this purpose, disease severity of the chosen genotypes in different growth stages was evaluated. The results indicated that the bell pepper genotypes respond differently to pathogenicity tests. Based on cluster analyses confirmed by the results of SAS analyses, bell pepper cultivars were categorised in five distinct groups.     

Evidence of genetically diverse virulent mating types of <Emphasis Type="Italic">Phytophthora capsici</Emphasis> from <Emphasis Type="Italic">Capsicum annum</Emphasis> L.

Chili pepper (Capsicum annum L.) is an important economic crop that is severely destroyed by the filamentous oomycete Phytophthora capsici. Little is known about this pathogen in key chili pepper farms in Punjab province, Pakistan. We investigated the genetic diversity of P. capsici strains using standard taxonomic and molecular tools, and characterized their colony growth patterns as well as their disease severity on chili pepper plants under the greenhouse conditions. Phylogenetic analysis based on ribosomal DNA (rDNA), β-tubulin and translation elongation factor 1α loci revealed divergent evolution in the population structure of P. capsici isolates. The mean oospore diameter of mating type A1 isolates was greater than that of mating type A2 isolates. We provide first evidence of an uneven distribution of highly virulent mating type A1 and A2 of P. capsici that are insensitive to mefenoxam, pyrimorph, dimethomorph, and azoxystrobin fungicides, and represent a risk factor that could ease outpacing the current P. capsici management strategies.     

Morphological, Pathological and Molecular Variability in Colletotrichum capsici, the Cause of Fruit Rot of Chillies in the Subtropical Region of North-western India

Fruit rot of chillies (Capsicum annuum L.), caused by Colletotrichum capsici under tropical and subtropical conditions, results in qualitative and quantitative yield losses. Based on variation in cultural and morphological traits of C. capsici populations, 37 isolates were categorized into five groups designated, respectively, as Cc‐I, Cc‐II, Cc‐III, Cc‐IV and Cc‐V. In culture, most of the isolates produced cottony, fluffy or suppressed colonies. However, no significant differences were noticed in shape and size of conidia. The reaction of the 37 isolates on an indigenously developed differential set of Capsicum cultivars indicated the existence of different virulences in Himachal Pradesh (HP) chilli populations. Fifteen pathotypes of the pathogen were characterized from various chilli‐growing regions of HP. Pathotype CCP‐1 was most virulent and attacked all the differential cultivars. The genetic relationship between five morphological groups recognized within C. capsici was investigated using random amplified polymorphic DNA (RAPD) analysis. Molecular polymorphism generated by RAPD confirmed the variation in virulences of C. capsici and different isolates were grouped into five clusters. However, four isolates (Cc‐5, Cc‐33, Cc‐29 and Cc‐37) exhibited identical RAPD haplotypes. The pathological and RAPD grouping of isolates suggested no correlation among the test isolates.     

A key QTL cluster is conserved among accessions and exhibits broad-spectrum resistance to Phytophthora capsici: a valuable locus for pepper breeding

Developing cultivars carrying effective resistance against destructive pathogens has become a priority for breeders. While little is currently known about the genetic basis of durable resistance, it is generally associated with polygenic and broad-spectrum resistance. In this study, we assessed the spectrum of resistance to Phytophthora capsici conferred by a major effect quantitative trait locus (QTL) that has been detected in all of the resistant pepper accessions studied to date. After adding new markers derived from tomato sequences and those from pepper reported in the literature to three maps of pepper chromosome P5, we detected a QTL cluster involved in P. capsici resistance. By means of meta-analyses, we determined the occurrence of these QTLs in different genetic backgrounds and with different P. capsici isolates. Comparative mapping with tomato and potato highlighted a complex mosaic of Phytophthora resistance loci on colinear chromosome segments. We tested different lines with and without one of these QTLs, Pc5.1, with four isolates that we determined to be genetically distinct. Our data demonstrate that Pc5.1 is active against 12 isolates from different geographical origins and that it is conserved in all of the resistant accessions tested. We propose that this QTL is a key element responsible for the broad-spectrum resistance to P. capsici and, therefore, is a valuable locus for improving the effective resistance of pepper to P. capsici.     

Investigation of the genetic diversity of Phytophthora capsici in China using a universal fluorescent labelling method

Phytophthora capsici is an important oomycete pathogen threatening the vegetable production in China, but very little is known about its population structure. The objective of the present study was to evaluate the genetic diversity of 49 P. capsici isolates obtained from 2007 to 2014 at nine provincial locations in China. Isolates were assessed for mating type, metalaxyl resistance and simple sequence repeat (SSR) genotype. Mating‐type analyses of the isolates showed that both mating types were present in all of the sampled production regions, and the mating‐type frequency in the total Chinese population did not deviate significantly from a 1:1 ratio. Responses of isolates to the fungicide metalaxyl indicated the presence of intermediate resistance to metalaxyl among the field population. A universal fluorescent labelling method was adapted in this study to improve the efficiency of SSR genotyping. Microsatellite genotyping of the isolates using seven SSR markers revealed 44 unique multilocus genotypes. Genetic analyses indicated the existence of two genetic clusters within Chinese P. capsici collection. Clonal reproduction may play a more prominent role in Yunnan Province, but non‐existence of repeated genotypes and existence of both mating types throughout all regions suggest outcrossing and sexual recombination likely play an important role in the overall epidemiology in China. Future studies would include expanded scale sampling at single regions over multiple years to better define the genetic diversity of P. capsici in China.     

Development of an Improved Isolation Approach and Simple Sequence Repeat Markers To Characterize Phytophthora capsici Populations in Irrigation Ponds in Southern Georgia

Phytophthora capsici, the causal agent of Phytophthora blight, is a major concern in vegetable production in Georgia and many other states in the United States. Contamination of irrigation water sources by P. capsici may be an important source of inoculum for the pathogen. A simple method was developed in this study to improve the efficiency of recovering P. capsici from fruits used as baits in irrigation ponds. In contrast to direct isolation on agar plates, infected fruit tissues were used to inoculate stems of pepper seedlings, and the infected pepper stems were used for isolation on agar plates. With isolation through inoculation of pepper stems, the frequency of recovering P. capsici from infected eggplant and pear fruits increased from 13.9% to 77.7% and 8.1% to 53.5%, respectively, compared with direct isolation on agar plates. P. capsici was isolated from seven out of nine irrigation ponds evaluated, with most of the ponds containing both A1 and A2 mating types and a 4:5 ratio of A1 to A2 when isolates from all ponds were calculated. All P. capsici isolates were pathogenic on squash plants, and only a small proportion (8.2%) of the isolates were resistant or intermediately sensitive to mefenoxam. Simple sequence repeats (SSRs) were identified through bioinformatics mining of 55,848 publicly available expressed sequence tags of P. capsici in dbEST GenBank. Thirty-one pairs of SSR primers were designed, and SSR analysis indicated that the 61 P. capsici isolates from irrigation ponds were genetically distinct. Cluster analysis separated the isolates into five genetic clusters with no more than two genetic groups in one pond, indicating relatively low P. capsici genetic diversity in each pond. The isolation method and SSR markers developed for P. capsici in this study could contribute to a more comprehensive understanding of the genetic diversity of this important pathogen.Phytophthora capsici, the causal agent of Phytophthora blight, is a widespread and destructive plant pathogen that causes root rot, crown rot, fruit rot, and foliar blight on many economically important crops in the United States and throughout the world (1). A number of important vegetable crops are susceptible to this pathogen, including peppers, squash, cucumber, watermelon, cantaloupe, zucchini, eggplant, pumpkin, tomatoes, and snap beans. The pathogen causes significant yield reductions and quality losses to vegetable industries and has become a major concern in vegetable production in the United States in recent years. The efficacies of current strategies for management of the disease are limited. No single fungicide has consistently and effectively suppressed losses caused by P. capsici epidemics. While fungicides containing the active ingredient mefenoxam provide some level of control of P. capsici, mefenoxam-resistant isolates that challenge the usefulness of the compound have developed (3, 8).It is critical to understand the ecology and epidemiology of P. capsici in order to design more effective disease management strategies. Studies conducted in recent years indicate that P. capsici survives in irrigation water in the United States, and irrigation water may serve as an important inoculum source. Roberts et al. (14) reported that P. capsici was isolated from tailwater (surface runoff water) in Florida using water filtration and lemon leaf baiting techniques. Gevens et al. (3) used pear and cucumber fruits as baits and isolated P. capsici from irrigation water sources in Michigan. It was unknown, however, if irrigation water sources in Georgia could be significant sources of primary inoculum. Earlier studies using water filtration or direct isolation from water and bottom sediment did not identify P. capsici in surface irrigation ponds in Georgia (16).Since surface water can be a significant source of P. capsici, it is critical to use appropriate methods to isolate the pathogen from irrigation water and to facilitate characterization of the isolates. Fruit, especially pears, is often used as bait to recover Phytophthora spp. from water (3, 21). In comparison to water filtration, the baiting technique is easier and less labor intensive. However, direct isolation from infected fruit bait is often hampered by other microorganisms. Isolation of Phytophthora spp. is often affected by Pythium spp. that overgrow fruit and agar plates. Hence, development of a more efficient isolation method is needed to increase the frequency of P. capsici recovery to facilitate the detection and characterization of isolates associated with water sources.The objectives of this study were to develop an efficient method to isolate P. capsici from irrigation ponds in southern Georgia and to develop simple sequence repeat (SSR) markers to analyze the genetic diversity of P. capsici populations in irrigation ponds. SSRs are tandemly repeated motifs of 1 to 6 bases found in the nuclear genomes of all eukaryotic organisms and are often abundant and evenly dispersed (7). They are highly polymorphic, multiallelic, and codominant and are believed to be a more efficient marker system than restriction fragment length polymorphisms and randomly amplified polymorphic DNAs (18, 23). SSR markers have been derived from publicly available expressed sequence tags (ESTs) of a few plant pathogens, including Phytophthora infestans, Phytophthora sojae, and Magnaporthe grisea (5, 10, 23); however, no SSRs for P. capsici have been developed. Development of EST-SSR markers may provide an effective molecular marker system for analysis of genetic variation within P. capsici populations.     

An investigation of biological attributes that may contribute to the importance of Phytophthora capsici as a vegetable pathogen in Florida

During the 1999–2000 and 2000–2001 seasons, 19 commercial squash fields in the vicinity of Homestead, Florida (USA) were examined for diseases caused by Phytophthora capsici. In each of the six fields in which two or more isolates of P. capsici were recovered, both the Al and A2 mating types were present, and both mating types were recovered from the same plant five times. Insensitivity to mefenoxam was common among isolates, with EC50s ranging from 5 μg mefenoxam ml^?1 to more than 60 μg ml^?1. Of 15 weed species that were examined as possible alternative hosts of the pathogen, only common purslane, Portulaca oleracea, was infected by P. capsici. Few or no oospores of the pathogen formed in a glasshouse (c. 28°C) when artificially inoculated pepper plants were covered with plastic bags or kept under continuous mist. In studies in the laboratory (c. 22°C) with detached pepper leaves, no oospores were formed on wire screens over water reservoirs. Consistent production of oospores occurred only when leaves were in constant contact with water. Maximum production occurred at 18°C, and production also occurred at 14°C, 20°C, 24°C and 26°C, but not at 6°C, 12°C, 30°C and 32°C.     

Plant-Plant-Microbe Mechanisms Involved in Soil-Borne Disease Suppression on a Maize and Pepper Intercropping System

Background

Intercropping systems could increase crop diversity and avoid vulnerability to biotic stresses. Most studies have shown that intercropping can provide relief to crops against wind-dispersed pathogens. However, there was limited data on how the practice of intercropping help crops against soil-borne Phytophthora disease.

Principal Findings

Compared to pepper monoculture, a large scale intercropping study of maize grown between pepper rows reduced disease levels of the soil-borne pepper Phytophthora blight. These reduced disease levels of Phytophthora in the intercropping system were correlated with the ability of maize plants to form a “root wall” that restricted the movement of Phytophthora capsici across rows. Experimentally, it was found that maize roots attracted the zoospores of P. capsici and then inhibited their growth. When maize plants were grown in close proximity to each other, the roots produced and secreted larger quantities of 2,4-dihydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one (DIMBOA) and 6-methoxy-2-benzoxazolinone (MBOA). Furthermore, MBOA, benzothiazole (BZO), and 2-(methylthio)-benzothiazole (MBZO) were identified in root exudates of maize and showed antimicrobial activity against P. capsici.

Conclusions

Maize could form a “root wall” to restrict the spread of P. capsici across rows in maize and pepper intercropping systems. Antimicrobe compounds secreted by maize root were one of the factors that resulted in the inhibition of P. capsici. These results provide new insights into plant-plant-microbe mechanisms involved in intercropping systems.     

Genetic diversity of <Emphasis Type="Italic">Phytophthora capsici</Emphasis> recovered from Massachusetts between 1997 and 2014

Phytophthora blight caused by Phytophthora capsici limits the production of cucurbits and peppers in the United States and is a growing threat to sustainable vegetable production in New England. Little is known about the genetic diversity of P. capsici in New England, and a total of 210 P. capsici isolates from 18 sites were genotyped using 46 single nucleotide polymorphism markers, revealing 85 unique and 34 repeated multi-locus genotypes. Both mating types were recovered from 7 of the 18 locations. Isolates with identical genotypes (clonal lineages) ranged from 2 to 16. Three clonal lineages were recovered from multiple sites within the same year, although none were recovered across multiple years. Bayesian clustering revealed individuals with a complex genetic cluster composition. This, coupled with a high outcrossing rate (mean t = 0.87) and no clear clustering in principal coordinates analysis, suggests outcrossing among the populations. Phylogenetic and genetic distance analysis indicate differentiation based on farm location and movement among farms may be infrequent. There was no obvious differentiation based on cucurbit, tomato or pepper hosts.     

Functional analysis of Pcipg2 from the straminopilous plant pathogen Phytophthora capsici

Phytophthora capsici causes serious diseases in numerous crop plants. Polygalacturonases (PGs) are cell wall‐degrading enzymes that play an important role in pathogenesis in straminopilous pathogens. To understand PGs as they relate to the virulence of P. capsici, Pcipg2 was identified from a genomic library of a highly virulent P. capsici strain. Pcipg2 was strongly expressed during symptom development after the inoculation of pepper leaves with P. capsici. The wild protein (PCIPGII) was obtained from the expression of pcipg2 and found that increasing activity of PGs in PCIPGII‐treated pepper leaves was consistent with increasing symptom development. Asp residues in active sites within pcipg2 affected PCIPGII activity or its virulence on pepper leaves. Results show that pcipg2 is an important gene among pcipg genes, and illustrate the benefit of analyzing mechanisms of pathogenicity during the period of host/parasite interaction. genesis 47:535–544, 2009. © 2009 Wiley‐Liss, Inc.     

Evaluation of Oospore Hyperparasites for the Control of Phytophthora Crown Rot of Pepper

Nine isolates of known oospore mycoparasites comprised of six actinomycetes (Actinoplanes missouriensis, A. philippinensis, A. utahensis, Amorphosporangium auranticolor, Ampullariella regularis, Spirillospora albida) and three fungi (Acremonium sp., Humicola fuscoatra, Verticillium chlamydosporium) were tested in the greenhouse for their ability to suppress or delay the onset of crown rot of pepper caused by Phytophthora capsici. Verticillium chlamydosporium applied as a root dip increased the number of healthy plants by more than 100% when peppers were transplanted into soil artificially infested with oospores of Phytophthora capsici, but not when peppers were transplanted into soil naturally infested with P. capsici. The other mycoparasites were ineffective in the greenhouse. All the mycoparasites tested parasitized oospores of P. capsici in vitro.     

Intraspecies diversity of Macrophomina phaseolina in Iran

To study the pathogenic and genetic diversity of the Macrophomina phaseolina in Iran, 52 isolates of the fungus were isolated from 24 host plants across the 14 Iranian provinces. All isolates were confirmed to the species based on the species-specific primers. The aggressiveness of M. phaseolina isolates was evaluated on the common bean. Based on the pathogenicity tests, M. phaseolina isolates from the different hosts displayed different levels of aggressiveness on the common beans. The results showed that there was significant variation in the aggressiveness of the pathogen; however, there was no distinct pattern of differentiation based on the host or geographical origin linked to the virulence of the isolates, as frequently theisolates from the same host or geographical origin had different levels of aggressiveness. Inter-simple sequence repeat (ISSR) markers were used to assess the genetic diversity of the fungus. The unweighted pair-group method, using arithmetic mean clustering of data, showed that isolates did not clearly differentiate to the specific group according to the host or geographical origins; however, usually the isolates from the same host or the same geographical origin tend to group nearly. Our results did not show a correlation between the genetic diversity based on the ISSR and pathogenic patterns on common bean in the greenhouse. Similar to the M. phaseolina populations in the other countries, the Iranian isolates were highly diverse based on the pathogenic and genotypic characteristics.     

Ultrastructure at the Host-Parasite Interface of Phytophthora capsici in Roots and Stems of Capsicum annuum

The infecting hyphae of Phytophthora capsici grew intercellularly in infected tissues of roots and stems of pepper (Capsicum annuum). The vascular tissues were not markedly disorganized even when heavily infected. Intercellularly growing hyphae penetrated the host cells by forming haustorium-like bodies. The consistent features of ultrastructural changes in infected tissues of pepper roots and stems were degeneration of cell organelles and dissolution of host cell walls. The cytoplasm detached from the cell wall aggregated abundantly around some haustorium-like bodies or the penetration sites of fungal hyphae. The host cell walls were palely stained, thinned and swollen, possibly being biochemically altered by the action of fungal macerating enzymes. Electron-dense, wall-like material was apposed on the outer wall of xylem vessel contacted by fungal hyphae. The infecting hyphae were also surrounded by granular, dark-staining cytoplasm. Characteristics of host cell responses to the invading P. capsici were the deposition of papilla-like material on host cell walls next to hyphae and the encasement of haustorium-like bodies with wall appositions.     

Loss of Heterozygosity Drives Clonal Diversity of Phytophthora capsici in China

Phytophthora capsici causes significant loss to pepper (Capsicum annum) in China and our goal was to develop single nucleotide polymorphism (SNP) markers for P. capsici and characterize genetic diversity nationwide. Eighteen isolates of P. capsici from locations worldwide were re-sequenced and candidate nuclear and mitochondrial SNPs identified. From 2006 to 2012, 276 isolates of P. capsici were recovered from 136 locations in 27 provinces and genotyped using 45 nuclear and 2 mitochondrial SNPs. There were two main mitochondrial haplotypes and 95 multi-locus genotypes (MLGs) identified. Genetic diversity was geographically structured with a high level of genotypic diversity in the north and on Hainan Island in the south, suggesting outcrossing contributes to diversity in these areas. The remaining areas of China are dominated by four clonal lineages that share mitochondrial haplotypes, are almost exclusively the A1 or A2 mating type and appear to exhibit extensive diversity based on loss of heterozygosity (LOH). Analysis of SNPs directly from infected peppers confirmed LOH in field populations. One clonal lineage is dominant throughout much of the country. The overall implications for long-lived genetically diverse clonal lineages amidst a widely dispersed sexual population are discussed.     

Pathogenic Variability and Genetic Diversity of Phaeoisariopsis griseola Isolates from Two Counties in the State of Goias, Brazil

Angular leaf spot disease of common bean (Phaseolus vulgaris), caused by Phaeoisariopsis griseola, is one of the most important disease of this crop in Brazil. Control strategies for the disease include cultural practices, chemical control and genetic resistance. This pathogen is known to vary greatly in pathogenicity. For durable use of genetic resistance to control this disease, it is necessary to manage resistant cultivars by taking into account the population structure of P. griseola. Isolates of the pathogen from Goias, Brazil exhibited an important virulence polymorphism when inoculated on 12 differential cultivars. A total of 13 pathotypes was identified within a series of 96 isolates collected in Inhumas and Damolandia counties. Only pathotypes 63‐15, 63‐23, 63‐31 and 63‐63 were identified in both counties. Since all the isolates were capable of inducing disease in both Andean and Mesoamerican differential cultivars, they were considered to be of Mesoamerican origin. Random amplified polymorphic DNA (RAPD) analysis performed on the same 96 isolates revealed a great genetic diversity clustering the series into five groups at an Euclidean distance of 62.5%. Although the results did not show any clustering according to the isolate origin, it was possible to observe a tendency of the isolates to cluster in different groups according to their origin. No pathotype‐specific band was observed in the present study.     

Host Specificity and Genetic Diversity of Didymella bryoniae from Cucurbitaceae in Brazil

Thirty‐seven isolates of Didymella bryoniae from three Cucurbitaceae species were collected in Brazil and tested for pathogenicity to watermelon. All isolates were pathogenic but differed in aggressiveness levels. Seven representative isolates were used in cross‐pathogenicity tests against 10 cucurbitaceous hosts. Most isolates were pathogenic to most host species tested, except to Sechium edule. Among the susceptible species, Citrullus and Cucumis species were the most susceptible hosts, while pumpkin and Luffa purgans were the most resistant. Host of origin affected the pattern of aggressiveness on each host. Isolates from watermelon were very aggressive to their original host, but much less aggressive or not pathogenic at all to some Cucurbita. Two previously described random‐amplified polymorphic DNA (RAPD)‐specific primers indicated that 81% of the isolates could be classified into the so‐called RG I group, while the remaining isolates could not be classified into any of the described RG groups. All 37 isolates were further characterized by RAPD fingerprinting and compared with three US isolates representative of RG I and RG II groups. The Brazilian D. bryoniae isolates could be separated into genetically similar clusters. The majority of the isolates were grouped in cluster DB Ia, which contained only isolates of Citrullus lanatus and Cucumis melo. Two of the American isolates used as controls clustered with this group at 68% similarity level. The DB Ib cluster included three Brazilian isolates obtained from melon and watermelon and the American representative for RG II, at a lower similarity level (43%). Two isolates from watermelon clustered with one isolate from melon in a separate group (DB II), while one single isolate from pumpkin (DB III) showed the lowest genetic similarity to all other isolates. Didymella bryoniae isolates from Brazil showed, therefore, a level of genetic diversity higher than previously reported for the species. RAPD fingerprinting allowed for geographical distinction of D. bryoniae isolates but no correlation between genetic distance, aggressiveness or origin of the isolate was found.     

Morphological and Pathogenic Characterization of Genetically Diverse Sclerotinia Isolates from European Red Clover Crops (Trifolium Pratense L.)

Clover rot, an important disease in European red clover crops, is caused by Sclerotinia trifoliorum or Sclerotinia sclerotiorum. Until today, little is known about the variation in aggressiveness among Sclerotinia isolates from red clover. Aggressiveness has never been correlated with morphological characteristics. Rapidly growing isolates may be more aggressive, but this was never investigated in S. trifoliorum before. Also nothing is known about the link between sclerotia production and aggressiveness. Oxalic acid is an important pathogenicity factor in Sclerotinia species, but its effect on aggressiveness is unknown in S. trifoliorum isolates. For this study, we selected 30 Sclerotinia isolates from 25 locations Europe: 26 S. trifoliorum isolates and 4 S. sclerotiorum isolates from two locations in France (Fr.A and Fr.B). For each isolate, the in vitro growth speed, sclerotia production, oxalate production and aggressiveness were analysed and correlations were estimated between aggressiveness and the other characteristics. Aggressiveness was assessed in vitro on detached leaves and in a greenhouse on young plants. Our isolates differed significantly in growth speed, sclerotia production, oxalate production and aggressiveness. The infections on detached leaves and young plants revealed interaction between isolates and plant genotypes and between isolates and cultivars, but there was no indication that pathotypes exist. In vitro growth speed and in vitro aggressiveness on detached leaves were positively correlated with aggressiveness on young plants, while sclerotia production was negatively correlated with aggressiveness on young plants. These factors can be used as predictors of aggressiveness of Sclerotinia isolates from red clover crops.     

Survival and spread of Phytophthora capsici on Long Island, New York

Phytophthora capsici is an oomycete soil-borne plant pathogen that causes root, fruit and foliar disease on a variety of vegetables. The epidemiology and population structure varies depending on the region surveyed and our objective was to investigate survival and spread on farms on Long Island, New York using single nucleotide polymorphism (SNP) markers. A total of 373 P. capsici isolates were collected from pumpkin, pepper, watermelon and snap bean on 15 farms. Both mating type were recovered from most locations. Genotypic analysis was conducted using 14 SNP loci located primarily within genes. A total of 128 unique multi-locus genotypes were identified. Of these, 54 appear to be clonal lineages ranging in size from 2 to 26 members. Most clonal lineages were recovered during the same year. Our results indicate that both sexual and clonal reproduction play important roles in the epidemiology of P. capsici on Long Island, NY, USA. The implications for managing the disease are discussed.     

Defence response of pepper (Capsicum annuum) suspension cells to Phytophthora capsici

Cell suspension cultures of three cultivars of Capsicum annuum L., with different degrees of sensibility to the fungus Phytophthora capsici, responded to elicitation by both lyophilized mycelium and fungus filtrate. They showed conductivity changes, browning, production of the phytoalexin capsidiol and synthesis or accumulation of pathogenesis-related (PR) proteins with glucanase (EC 3.2.1.39) and chitinase (EC 3.2.1.14) activities. The cultivation medium was optimised for growth of both the plant and the fungus in order to avoid any stress during their combination. The resistant cv. Smith-5, showed a more rapid and intense response to the elicitor preparations than the sensitive cvs Americano and Yolo Wonder. This was particularly evident when the cell suspensions were elicited with the filtrate, when differences became clearly visible after only 6 h incubation. The greatest rate of capsidiol accumulation occurred after 18 h in the mycelium-elicited cells and after 12 h in those elicited with the filtrate. These times are the optimal for capsidiol accumulation, and the phytoalexin is produced much more rapidly than it can be excreted into the extracellular medium. The inhibition threshold of fungal growth (300 µg capsidiol [g dry weight]^?1) was reached only in the resistant cultivar. The induction of an intracellular glucanase (pI 8.9 and R_f 0.18) and an extracellular chitinase (pI 5.4 and R_f 0.70) only in the resistant cultivar 24 h after elicitation suggests that these enzymes are involved in the resistance to Phytophthora capsici, while other hydrolases common to all three cultivars form part of a more general defence. The results indicate that elicitation of pepper cell suspension cultures by signal molecules from P. capsici exhibits properties of a multicomponent dynamic system in which different protective mechanisms play complementary roles in the overall expression of the defence reaction. We confirm that the differential responses of resistant and susceptible pepper cultivars to P. capsici previously seen in plant stem sections are retained in suspension culture.