Sporulation of Fusarium oxysporum f. sp. lycopersici on Stem Surfaces of Tomato Plants and Aerial Dissemination of Inoculum

ABSTRACT Plants exhibiting symptoms of wilt and xylem discoloration typical of Fusarium wilt caused by Fusarium oxysporum f. sp. lycopersici were observed in greenhouses of cherry tomatoes at various sites in Israel. However, the lower stems of some of these plants were covered with a pink layer of macroconidia of F. oxysporum. This sign resembles the sporulating layer on stems of tomato plants infected with F. oxysporum f. sp. radicis-lycopersici, which causes the crown and root rot disease. Monoconidial isolates of F. oxysporum from diseased plants were assigned to vegetative compatibility group 0030 of F. oxysporum f. sp. lycopersici and identified as belonging to race 1 of F. oxysporum f. sp. lycopersici. The possibility of coinfection with F. oxysporum f. sp. lycopersici and F. oxysporum f. sp. radicis-lycopersici was excluded by testing several macroconidia from each plant. Airborne propagules of F. oxysporum f. sp. lycopersici were trapped on selective medium in greenhouses in which plants with a sporulating layer had been growing. Sporulation on stems was reproduced by inoculating tomato plants with races 1 and 2 of F. oxysporum f. sp. lycopersici. This phenomenon has not been reported previously with F. oxysporum f. sp. lycopersici and might be connected to specific environmental conditions, e.g., high humidity. The sporulation of F. oxysporum f. sp. lycopersici on plant stems and the resultant aerial dissemination of macroconidia may have serious epidemiological consequences. Sanitation of the greenhouse structure, as part of a holistic disease management approach, is necessary to ensure effective disease control.     

Disease Development Following Infection of Tomato and Basil Foliage by Airborne Conidia of the Soilborne Pathogens Fusarium oxysporum f. sp. radicis-lycopersici and F. oxysporum f. sp. basilici

ABSTRACT Fusarium oxysporum f. sp. radicis-lycopersici, the causal agent of Fusarium crown and root rot of tomato, and F. oxysporum f. sp. basilici, the causal agent of Fusarium wilt in basil, are soilborne pathogens capable of producing conspicuous masses of macroconidia along the stem. The role of the airborne propagules in the epidemics of the disease in tomato plants was studied. In the field, airborne propagules of F. oxysporum f. sp. radicis-lycopersici were trapped with a selective medium and their prevalence was determined. Plants grown in both covered and uncovered pots, detached from the field soil, and exposed to natural aerial inoculum developed typical symptoms (82 to 87% diseased plants). The distribution of inoculum in the growth medium in the pots also indicated the occurrence of foliage infection. In greenhouse, foliage and root inoculations were carried out with both tomato and basil and their respective pathogens. Temperature and duration of high relative humidity affected rate of colonization of tomato, but not of basil, by the respective pathogens. Disease incidence in foliage-inoculated plants reached 75 to 100%. In these plants, downward movement of the pathogens from the foliage to the crown and roots was observed. Wounding enhanced pathogen invasion and establishment in the foliage-inoculated plants. The sporulation of the two pathogens on stems, aerial dissemination, and foliage infection raise the need for foliage protection in addition to soil disinfestation, in the framework of an integrated disease management program.     

Treatment with the Mycoparasite Pythium oligandrum Triggers Induction of Defense-Related Reactions in Tomato Roots When Challenged with Fusarium oxysporum f. sp. radicis-lycopersici

ABSTRACT The influence exerted by the mycoparasite Pythium oligandrum in triggering plant defense reactions was investigated using an experimental system in which tomato plants were infected with the crown and root rot pathogen Fusarium oxysporum f. sp. radicis-lycopersici. To assess the antagonistic potential of P. oligandrum against F. oxysporum f. sp. radicis-lycopersici, the interaction between the two fungi was studied by scanning and transmission electron microscopy (SEM and TEM, respectively). SEM investigations of the interaction region between the fungi demonstrated that collapse and loss of turgor of F. oxysporum f. sp. radicis-lycopersici hyphae began soon after close contact was established with P. oligandrum. Ultrastructural observations confirmed that intimate contact between hyphae of P. oligandrum and cells of the pathogen resulted in a series of disturbances, including generalized disorganization of the host cytoplasm, retraction of the plasmalemma, and, finally, complete loss of the protoplasm. Cytochemical labeling of chitin with wheat germ agglutinin (WGA)/ovomucoid-gold complex showed that, except in the area of hyphal penetration, the chitin component of the host cell walls was structurally preserved at a time when the host cytoplasm had undergone complete disorganization. Interestingly, the same antagonistic process was observed in planta. The specific labeling patterns obtained with the exoglucanase-gold and WGA-ovomucoid-gold complexes confirmed that P. oligandrum successfully penetrated invading cells of the pathogen without causing substantial cell wall alterations, shown by the intense labeling of chitin. Cytological investigations of samples from P. oligandrum-inoculated tomato roots revealed that the fungus was able to colonize root tissues without inducing extensive cell damage. However, there was a novel finding concerning the structural alteration of the invading hyphae, evidenced by the frequent occurrence of empty fungal shells in root tissues. Pythium ingress in root tissues was associated with host metabolic changes, culminating in the elaboration of structural barriers at sites of potential fungal penetration. Striking differences in the extent of F. oxysporum f. sp. radicis-lycopersici colonization were observed between P. oligandrum-inoculated and control tomato plants. In control roots, the pathogen multiplied abundantly through much of the tissues, whereas in P. oligandrum-colonized roots pathogen growth was restricted to the outermost root tissues. This restricted pattern of pathogen colonization was accompanied by deposition of newly formed barriers beyond the infection sites. These host reactions appeared to be amplified compared to those seen in nonchallenged P. oligandrum-infected plants. Most hyphae of the pathogen that penetrated the epidermis exhibited considerable changes. Wall appositions contained large amounts of callose, in addition to be infiltrated with phenolic compounds. The labeling pattern obtained with gold-complexed laccase showed that phenolics were widely distributed in Fusarium-challenged P. oligandrum-inoculated tomato roots. Such compounds accumulated in the host cell walls and intercellular spaces. The wall-bound chitin component in Fusarium hyphae colonizing P. oligandrum-inoculated roots was preserved at a time when hyphae had undergone substantial degradation. These observations provide the first convincing evidence that P. oligandrum has the potential to induce plant defense reactions in addition to acting as a mycoparasite.     

Fusarium crown and root rot of tomatoes in the UK

Fusarium crown and root rot caused by Fusarium oxysporum f. sp. radicis-lycopersici was found in the UK in 1988 and 1989 mainly in rockwool-grown tomato crops. Up to 14% of plants were affected in individual crops. In experiments, leaf and stem symptoms did not appear until the time of first fruit harvest even when the plants were inoculated at planting, first flowers or fruit set. Conidial inoculum at 10⁶ spores/plant applied at seed sowing killed 70–83% of tomato seedlings, whereas similar levels of inoculum applied to young plants caused root and basal stem decay, and eventually death but only after fruit harvest began. Disease incidence and symptom severity increased with inoculum concentration. Experimentally, the disease was more severe in peat- or compost-grown plants than in rockwool. Disease spread was only a few centimetres in 50 days in experimental rockwool-grown plants. All tomato cultivars tested were highly susceptible. Prochloraz-Mn was highly effective against the pathogen in vitro and controlled the disease in the glasshouse, but only when applied preventively. Non-pathogenic Fusarium oxysporum isolates and Trichoderma harzianum also reduced FCRR disease levels.     

The infestation of tomato seed by Fusarium oxysporum f.sp. radicis-lycopersici

The infestation of seed by Fusarium oxysporum f.sp. radicis-lycopersici occurred at rates of 0-1 to 0-01 % in fruit on stem-infected plants. Direct infection of fruit in the flower or young developing fruit stage resulted in a grayish-brown lesion on the stylar end of the fruit or mummification. F. oxysporum f.sp. radicis-lycopersici was isolated from all seeds in such fruit. Picked fruit inoculated on the stem scar also became infected but 96 h after inoculation of the fruit, the seed was not infested or infected. The contact of clean seed with hands that had previously handled F. oxysporum f.sp. radicis-lycoperisid-infesied sawdust resulted in a high level of seed infestation. The fungus survived on seed sent across Canada and stored for up to 12 weeks. Treatment with NaOCl or 0·1 N HCl did not completely disinfest infested seed.     

The genetic analysis of resistance to fusarium crown and root rot of tomato

The tomato line IRB-301-31, resistant to fusarium crown and root rot ( Fusarium oxysporum f.sp. radicis-lycopersici ) was crossed with two susceptible cultivars, Motelle and Earlypak No. 7. When F₁, F₂ and backcross progenies were inoculated at the one-leaf stage with a suspension of spores of the pathogen, all could be classified as either resistant (healthy) or susceptible (dead). The ratios of resistant to susceptible plants indicated that resistance was conferred by a single dominant gene, designated Fr1.     

Use of a Nitrate-Nonutilizing Mutant and Selective Media to Examine Population Dynamics of Fusarium oxysporum f. sp. spinaciae in Soil

ABSTRACT Determining the population density of the spinach wilt pathogen Fusarium oxysporum f. sp. spinaciae in soil with conventional Fusarium-selective media is quite difficult because nonpathogenic strains of F. oxysporum also grow on those media and are indistinguishable from the pathogen. Therefore, a nitrate-nonutilizing (nit) mutant of the pathogen and corresponding selective media were tested in an experimental approach to determine the population density of the pathogen. Colony forming units of the pathogen were countable after soil-dilution plating onto nit mutant-selective media MMCPA, CMP, and CGMBP. Colony forming units of wild-type Fusarium spp. were countable using a wildtype Fusarium-selective medium, GMBP. By combining nit mutant- and wild-type-selective media, the population densities of pathogenic and nonpathogenic F. oxysporum in the same soil could be measured selectively. This method was useful in studying population dynamics of the pathogen after different soil treatments. Soil disinfested with hot water or chloropicrin was amended with the nit mutant pathogen, and subsequent changes in population densities of the pathogen were compared with those in nontreated field soil. The pathogen rapidly proliferated in disinfested soil and wilt developed faster than in nontreated soil. When a nonpathogenic isolate of F. oxysporum was added at high density to sterilized soil prior to the pathogen, growth of the pathogen was greatly suppressed. Nonpathogenic F. oxysporum could not, however, reduce the density of preexisting pathogen.     

Combined soil treatments and sequence of application in improving the control of soilborne pathogens

ABSTRACT The effects of reduced doses of methyl bromide (MB) or metham sodium, heating, short solarization, and soil microbial activity, alone or in combination, on survival of soilborne fungal pathogens were tested in a controlled-environment system and field plots. Sublethal doses of heating or MB delayed germination of Sclerotium rolfsii sclerotia. Combining MB and heating treatments was more effective than either treatment alone in controlling S. rolfsii and Fusarium oxysporum f. sp. basilici. The application heating followed by fumigation with MB, was significantly more effective in delaying and reducing germination of S. rolfsii sclerotia and in controlling F. oxysporum f. sp. basilici than the opposite sequence. Further, incubation in soil and exposure to microbial activity of previously heated or MB-treated sclerotia increased the mortality rate, indicating a weakening effect. Similarly, incubation of chlamydospores of F. oxysporum f. sp. melonis and F. oxysporum f. sp. radicis-lycopersici in soil in the field after fumigation further reduced their survival, confirming the laboratory results. In field tests, combining MB or metham sodium at reduced doses with short solarization was more effective in controlling fungal pathogens than either treatment alone. Treatment sequence significantly affected pathogen control in the field, similar to its effect under controlled conditions. This study demonstrates a frequent synergistic effect of combining soil treatments and its potential for improving pathogen control and reducing pesticide dose, especially when an appropriate sequence was followed.     

Vegetative compatibility and heterokaryon stability in Fusarium oxysporum f.sp. radicis-lycopersici from Italy

Fusarium crown and root rot, caused by Fusarium oxysporum f.sp. radicis-lycopersici ( Forl ), is one of the most destructive soilborne diseases of tomato in Italy. Chlorate-resistant, nitrate-nonutilizing ( nit ) mutants were used to determine vegetative compatibility among 191 isolates of Forl collected in five geographic regions (Calabria, Emilia-Romagna, Liguria, Sardinia, Sicily) in Italy. The isolates were assigned to five vegetative compatibility groups (VCGs): 65 isolates to VCG 0090; 99 to VCG 0091; 23 to VCG 0092; two to VCG 0093; and two to VCG 0096. The population structure of Forl in Italy is similar to that reported for Israel, and differs from that found in North Atlantic European countries, where VCG 0094 is predominant. The stability of prototrophic heterokaryons originating from hyphal anastomosis between compatible complementary nit mutants was assessed through conidial analysis and mycelial mass transfer. Most monoconidial cultures (84%) recovered from 117 prototrophic heterokaryons were nit mutants, indicating that heterokaryons generally do not proliferate well through conidiation; most of the 177 prototrophic heterokaryons examined were unstable, and only 9% sustained prototrophic growth through the tenth mycelial transfer upon subculturing. The prototrophic growth is proposed to be maintained through restoration of the heterokaryotic state by continual anastomosis between adjacent homokaryotic hyphae. Since heterokaryosis is a prerequisite for parasexual recombination, we speculate that this mechanism is unlikely to play a major role in generating the VCG diversity found among Forl or other strains of F. oxysporum.     

Plant defence reactions against fusarium wilt in chickpea induced by incompatible race 0 of Fusarium oxysporum f.sp. ciceris and nonhost isolates of F. oxysporum

Germinated seeds of 'kabuli' chickpea cv. ICCV 4 were inoculated with a conidial suspension of the incompatible race 0 of Fusarium oxysporum f.sp. ciceris (Foc) or of nonhost F. oxysporum resistance 'inducers', and 3 days later were challenged by root dip with a conidial suspension of highly virulent Foc race 5. Prior inoculation with inducers delayed the onset of symptoms and/or significantly reduced the final amount of fusarium wilt caused by race 5. However, the extent of disease suppression varied with the nature of the inducing agent; the nonhost isolates of F. oxysporum were more effective at disease suppression than the incompatible Foc race 0. Inoculation with the inducers gave rise to synthesis of maackiain and medicarpin phytoalexins in inoculated seedlings; these did not accumulate in plant tissues but were released into the inoculum suspension. Inoculation with inducers also resulted in accumulation of chitinase, β-1,3-glucanase and peroxidase activities in plant roots. These defence-related responses were induced more consistently and intensely by nonhost isolates of F. oxysporum than by incompatible Foc race 0. The phytoalexins and, to a lesser extent, the antifungal hydrolases, were also induced after challenge inoculation with Foc race 5. However, in this case the defence responses were induced in both preinduced and noninduced plants infected by the pathogen. It is concluded that the suppression of fusarium wilt in this study possibly involved an inhibitory effect on the pathogen of preinduced plant defences, rather than an increase in the expression of defence mechanisms of preinduced plants following a subsequent challenge inoculation.     

An ultrastructural study of tomato roots inoculated with pathogenic and non-pathogenic necrotrophic fungi and a saprophytic fungus

Tomato cultivar Moneymaker was independently inoculated with Alternaria alternata, Cunninghamella elegans, Fusarium culmorum, F. oxysporum f.sp. lycopersici, F. oxysporum f.sp. pisi and Stromatinia gladioli and analysed ultrastructurally. The extent and amount of superficial fungal growth on tomato roots was similar but C. elegans , a saprophyte, was exceptional in that hyphae were not closely appressed to plant surfaces and did not adhere to plant cell walls.
In general, the type of plant responses to fungal colonization and infection were similar in all of the interactions studied, with the exception of C. elegans which did not infect tomato root tissue. The failure to penetrate tomato roots by C. elegans may have been associated with the lack of hyphal adhesion to plant cell walls. Migration of cytoplasm and wall apposition/penetration papilla formation were regularly observed in tomato root tissue beneath appressed hyphae and at sites of fungal infection. Specific cellular reactions in the exodermis, namely the formation of wall 'inclusions' and appearance of 'sensitive' cells, indicated that exodermal cells were particularly responsive to fungal challenge.
Fusarium oxysporum f.sp. lycopersici , a pathogen of tomato, invaded tomato root tissue more extensively than the other fungi inoculated onto tomato roots. Infection of tomato by the other fungi studied was variable, and the extent and success of fungal invasion was tentatively associated with their necrotrophic capability and typical host range.     

Prevalence and pathogenicity of spinach root pathogens of the genera Aphanomyces, Phytophthora, Fusarium, Cylindrocarpon, and Rhizoctonia in Sweden

In a 4-year disease survey in commercial spinach fields, pathogens were isolated from spinach root pieces placed on selective agar media. Aphanomyces cladogamus was the most abundant pathogen, followed by Phytophthora. cryptogea and Fusarium oxysporum. Rhizoctonia solani was found only occasionally. Other pathogens isolated were F. redolens, F. sambucinum and Cylindrocarpon destructans. P. cryptogea was the most severe pathogen, causing death of most plants, but A. cladogamus also caused severe root damage. Isolates of F. oxysporum ranged from highly pathogenic, i.e. P. oxysporum f.sp. spinaciae race 1. to moderately pathogenic and non-pathogenic, Rhizoctonia solani isolates also varied widely in their pathogenicity. Only a small number of the F. redotens and F. sambucinum isolates were pathogenic and most C. destructans isolates were weakly pathogenic. Isolation frequencies were relatively stable from year to year, but P. cryptogea was isolated more frequently in autumn than in spring. No clear relationships were found between pathogen prevalence and disease severity index of surveyed field plants, between pathogen prevalence and plant developmental stage, or between prevalence of the different pathogens isolated.     

In planta and soil quantification of Fusarium oxysporum f. sp. ciceris and evaluation of Fusarium wilt resistance in chickpea with a newly developed quantitative polymerase chain reaction assay

Fusarium wilt of chickpea caused by Fusarium oxysporum f. sp. ciceris can be managed by risk assessment and use of resistant cultivars. A reliable method for the detection and quantification of F. oxysporum f. sp. ciceris in soil and chickpea tissues would contribute much to implementation of those disease management strategies. In this study, we developed a real-time quantitative polymerase chain reaction (q-PCR) protocol that allows quantifying F. oxysporum f. sp. ciceris DNA down to 1 pg in soil, as well as in the plant root and stem. Use of the q-PCR protocol allowed quantifying as low as 45 colony forming units of F. oxysporum f. sp. ciceris per gram of dry soil from a field plot infested with several races of the pathogen. Moreover, the q-PCR protocol clearly differentiated susceptible from resistant chickpea reactions to the pathogen at 15 days after sowing in artificially infested soil, as well as the degree of virulence between two F. oxysporum f. sp. ciceris races. Also, the protocol detected early asymptomatic root infections and distinguished significant differences in the level of resistance of 12 chickpea cultivars that grew in that same field plot infested with several races of the pathogen. Use of this protocol for fast, reliable, and cost-effective quantification of F. oxysporum f. sp. ciceris in asymptomatic chickpea tissues at early stages of the infection process can be of great value for chickpea breeders and for epidemiological studies in growth chambers, greenhouses and field-scale plots.     

Development of crown and root rot disease of tomato under irrigation with saline water

ABSTRACT We studied the effect of water salinity on the incidence and severity of crown and root rot disease of tomato, as well as on the pathogen and on the plant's response to the pathogen. Irrigation with saline water significantly increased disease severity in tomato transplants inoculated with Fusarium oxysporum f. sp. radicis-lycopersici, and mineral fertilization further increased it. In one field experiment, disease incidence in plots irrigated with saline water (electrical conductivity [EC] = 3.2 +/- 0.1 dS m(-1)) and in those irrigated with fresh water (EC = 0.4 +/- 0.1 dS m(-1)) was 75 and 38%, respectively. Disease onset was earlier and yield was lower in plots irrigated with saline water. In a second field experiment, final disease incidence 250 days after planting, was 12% in plants which had been irrigated with saline water (EC = 4.6 +/- 0.1 dS m(-1)) and 4% in those irrigated with fresh water (EC = 1.2 +/- 0.1 dS m(-1)). Irrigation of tomato transplants with 20 mM NaCl did not inhibit plant development, but partial inhibition was observed at higher NaCl concentrations. Growth of the pathogen in culture or survival of conidia added to soil were not affected by saline water. Plants which were preirrigated with saline water were more severely diseased than those preirrigated with tap water. It was concluded that disease increases effected by saline water are associated with the latter's effect on plant response.     

Infection by Meloidogyne artiellia does not break down resistance to races 0, 1a, and 2 of Fusarium oxysporum f. sp. ciceris in chickpea genotypes

Fusarium oxysporum f. sp. ciceris, and the root-knot nematode Meloidogyne artiellia, coinfect chickpea crops in several countries of the Mediterranean Basin. The influence of root infection by M. artiellia on the reactions of chickpea genotypes with different reaction to infection with F. oxysporum f. sp. ciceris races 0, 1A, and 2 was investigated under controlled environmental conditions. Results demonstrated that co-infection of chickpea genotypes resistant to specific fungal races by M. artiellia did not influence the Fusarium wilt reaction of the plant, irrespective of the F. oxysporum f. sp. ciceris race assayed. However, in some of the assayed combinations, coinfection by both pathogens significantly affected the level of colonization by the fungus or reproduction of the nematode in the root system. Thus, coinfection of chickpea plants with Foc-0 and M. artiellia significantly decreased the level of colonization of the root system by F. oxysporum f. sp. ciceris in genotypes 'CA 336.14.3.0' and 'PV 61', but not in 'ICC 14216 K' and 'UC 27'. Similarly, the nematode reproduction index was also significantly reduced by coinfection with Foc-0 in the four chickpea genotypes tested and inoculated with this race. Conversely, coinfection of chickpea plants with Foc-1A and M. artiellia significantly increased colonization of the root system by the fungus in all genotypes inoculated with this race, except for line BG 212. Altogether, we confirmed the complete resistance phenotype of 'UC 27' and 'ICC 14216 K' to Foc-0, and of 'ICC 14216 K' to Foc-1A and Foc-2, and demonstrated that this resistance was not modified by coinfection of the resistant plant with M. artiellia.     

Differential Colonization of Tomato Roots by Nonpathogenic and Pathogenic Fusarium oxysporum Strains May Influence Fusarium Wilt Control

ABSTRACT Histochemical staining, beta-glucuronidase (GUS) activity, or placing roots on agar were methods used to characterize interactions between the pathogenic fungus, Fusarium oxysporum f. sp. lycopersici, and the nonpathogenic biocontrol F. oxysporum strain 70T01 with respect to colonization behaviors, interaction sites, and population densities on tomato roots. Mycelia of strain 70T01, a genetic transformant expressing stable GUS activity, hygromycin B resistance, and effective disease control, were localized in epidermal and cortex cell layers of tomato roots in a discontinuous and uneven pattern. In contrast, mycelia of F. oxysporum f. sp. lycopersici were found in the vascular bundles. Thus, direct interactions between the two fungi likely happen in the root surface cell layers. Colonization density of strain 70T01 was related to the inoculation density but decreased with distance from the inoculation site. Host defense reactions, including increased cell wall thickness or papilla deposits, were adjacent to 70T01 hyphae. Experiments done in soil showed that strain 70T01 densities in roots were highest at inoculation zones and barely detectable for root segments more than 2 cm away from the inoculation sites. F. oxysporum f. sp. lycopersici densities were lowest at 70T01 inoculation zones and highest (>10 times) where strain 70T01 was not directly applied. Newly elongating roots where strain 70T01 did not reach were available for infection by the pathogen. The higher strain 70T01 density was always found when the plants were simultaneously infected by F. oxysporum f. sp. lycopersici, suggesting that F. oxysporum f. sp. lycopersici has as much influence in predisposing the plant to colonization by strain 70T01 as strain 70T01 has on providing disease protection against the pathogen.     

Modeling the survival of two soilborne pathogens under dry structural solarization

ABSTRACT Structural (space) solarization of a closed, empty greenhouse for sanitation involves dry heating to 60 degrees C and higher and low relative humidity (RH), under a fluctuating temperature and RH regime. Survival of inocula of Fusarium oxysporum f. sp. radicis-lycopersici and Sclerotium rolfsii during structural solarization was studied for 4 years (total of 12 experiments) in an attempt to develop a dynamic model for expressing the thermal inactivation of the pathogens. After 20 days of exposure, the populations of F. oxysporum f. sp. radicis-lycopersici and S. rolfsii were reduced by 69 to 95% and by 47.5 to 100%, respectively. The Weibull distribution model was applied to describe pathogen survival. The Weibull rate parameter, b, was found to follow an exponential (for F. oxysporum f. sp. radicis-lycopersici) and the Fermi (for S. rolfsii) functions at constant temperatures. To improve the applicability of the model, fluctuating conditions of both temperature and RH were utilized. The Weibull distribution derivative, expressed as a function of temperature and moisture, was numerically integrated to estimate survival of inocula exposed to structural solarization. Deviations between experimental and calculated values derived from the model were quite small and the coefficient of determination (R (2)) values ranged from 0.83 to 0.99 in 9 of 12 experiments, indicating that ambient RH data should be considered. Structural solarization for sanitation could be a viable component in integrated pest management programs.     

Characterization of Fusarium oxysporum f. sp. radicis-cucumerinum attacking melon under natural conditions in Greece

A severe root and stem rot disease of melon was observed during the 2001 growing season on four glasshouse crops in Heraklio, Greece. A total of 43 isolates of F. oxysporum , obtained in Crete from glasshouse-grown melon and showing fusarium wilt or root and stem rot symptoms, were characterized by pathogenicity and vegetative compatibility. The majority of these isolates was also fingerprinted via amplified fragment length polymorphic (AFLP) analysis. Of the total number of isolates, 22 were identified by pathogenicity tests as F. oxysporum f. sp. melonis , 20 as F. oxysporum f. sp. radicis-cucumerinum , while one isolate was nonpathogenic on cucumber, melon, sponge gourd and pumpkin. All 22 isolates of F. oxysporum f. sp. melonis were assigned to vegetative compatibility group (VCG) 0134, and all 20 isolates of F. oxysporum f. sp. radicis-cucumerinum to VCG 0260. Isolates of F. oxysporum f. sp. radicis-cucumerinum were incompatible with isolates of F. oxysporum f. sp. melonis. AFLP fingerprinting allowed for the clustering of the isolates of the two formae speciales of F. oxysporum along two separate phenetic groups: f. sp. melonis to AFLP major haplotype I, and f. sp. radicis-cucumerinum to AFLP major haplotype II. Overall, pathogenicity, vegetative compatibility grouping and AFLP analysis were correlated and effectively distinguished isolates of F. oxysporum from melon. This appears to be the first report of natural infection of melon by F. oxysporum f. sp. radicis-cucumerinum worldwide.     

Fungi on roots and stem bases of asparagus in the Netherlands: species and pathogenicity

A survey was made to identify the most important soilborne fungal pathogens of asparagus crops in the Netherlands. Ten plants were selected from each of five fields with a young (1–4 y) first planting, five fields with an old (6–13 y) first planting and five fields with a young replanting. The analysis included fungi present in the stem base and the roots of plants with symptoms of foot and root rot or showing growth decline without specific disease symptoms. Isolates of each species were tested for pathogenicity to asparagus on aseptically grown plantlets on Knop's agar. Symptoms were caused byFusarium oxysporum, F. culmorum, Botrytis cinerea, Penicillium verrucosum var.cyclopium, Cylindrocarpon didymum, Phialophora malorum, Phoma terrestris andAcremonium strictum. F. oxysporum was by far the most common species and was isolated from 80% of the plants. Not all of its isolates were pathogenic to asparagus. Symptoms were caused by 67%, 78% and 93% of the isolates obtained from young first plantings, old first plantings and replantings, respectively.F. culmorum was isolated from 31% of the plants. Two other notorious pathogens of asparagus,F. moniliforme andF. proliferatum, did not occur in our samples.Species causing symptoms in the vitro test that were found on more than 5% of the plants were additionally tested for their pathogenicity in pot experiments.F. oxysporum f.sp.asparagi caused severe foot and root rot, significantly reduced root weights and killed most of the plants.F. culmorum caused lesions on the stem base often resulting in death of the plant.P. terrestris, a fungus only once reported as a pathogen of asparagus, caused an extensive root rot, mainly of secondary roots that became reddish. The fungus was isolated in only a few samples and is not to be regarded as an important pathogen in Dutch asparagus crops.P. malorum caused many small brown lesions on the stem base and incidentally also on the upper part of small main roots. This is the first report of its pathogenicity to asparagus. The fungus is one of the organisms inciting spear rust and it reduced crop quality rather than crop yield.P. verrucosum var.cyclopium andC. didymum did not cause symptoms in pot experiments.Because of its predominance on plants with foot and root rot and its high virulence,F. oxysporum f.sp.asparagi was considered to be the main soilborne pathogen of asparagus in the Netherlands.     

Ultrastructural and Cytochemical Aspects of the Interaction Between the Mycoparasite Pythium oligandrum and Soilborne Plant Pathogens

ABSTRACT The interaction between the oomycete Pythium oligandrum and various soilborne oomycete and fungal plant pathogens (P. ultimum, P. aphanidermatum, Fusarium oxysporum f. sp. radicis-lycopersici, Verticillium albo-atrum, Rhizoctonia solani, and Phytophthora megasperma) was studied by light and electron microscopy in order to assess the relative contribution of mycoparasitism and antibiosis in the antagonistic process. Scanning electron microscope investigations of the interaction regions showed that structural alterations of all pathogenic fungi and oomycetes (except for Phytophthora megasperma) occurred soon after contact with the antagonist. Light and transmission electron microscope studies of the interaction region between the antagonist and P. ultimum revealed that intimate contact between both partners preceded a sequence of degradation events including aggregation of host cytoplasm and penetration of altered host hyphae. Localization of the host wall cellulose component showed that cellulose was altered at potential penetration sites. A similar scheme of events was observed during the interaction between P. oligandrum and F. oxysporum f. sp. radicis-lycopersici, with the exception that complete loss of host protoplasm was associated with antagonist invasion. The interaction between P. oligandrum and R. solani resulted in an abnormal deposition of a wall-like material at potential penetration sites for the antagonist. However, the antagonist displayed the ability to circumvent this barrier and penetrate host hyphae by locally altering the chitin component of the host hyphal wall. Interestingly, antagonist cells also showed extensive alteration as evidenced by the frequent occurrence of empty hyphal shells. In the case of Phytophthora megasperma, hyphal interactions did not occur, but hyphae of the plant pathogen were damaged severely. At least two distinct mechanisms appear to be involved in the process of oomycete and fungal attack by P. oligandrum: (i) mycoparasitism, mediated by intimate hyphal interactions, and (ii) antibiosis, with alteration of the host hyphae prior to contact with the antagonist. However, the possibility that the antagonistic process may rely on the dual action of antibiotics and hydrolytic enzymes is discussed.