Interactions Between Meloidogyne artiellia, the Cereal and Legume Root-Knot Nematode, and Fusarium oxysporum f. sp. ciceris Race 5 in Chickpea

ABSTRACT In the Mediterranean Basin, Fusarium oxysporum f. sp. ciceris and the root-knot nematode Meloidogyne artiellia coinfect chickpea. The influence of root infection (after inoculation with 20 nematode eggs and second-stage juveniles per gram of soil) by two M. artiellia populations, from Italy and Syria, on the reaction of chickpea lines and cultivars with partial resistance to Fusarium wilt (CA 252.10.1.OM, CA 255.2.5.0, CPS 1, and PV 61) and with complete resistance to F. oxysporum f. sp. ciceris race 5 (CA 334.20.4, CA 336.14.3.0, ICC 14216 K, and UC 27) was investigated under controlled conditions. In genotypes with partial resistance, infection by M. artiellia significantly increased the severity of Fusarium wilt, irrespective of the fungal inoculum density (3,000 or 30,000 chlamydospores per gram of soil), except in cultivar CPS 1 at the lower fungal inoculum density. In genotypes with complete resistance to Fusarium wilt, infection by M. artiellia overcame the resistance to F. oxysporum f. sp. ciceris race 5 in CA 334.20.4 and CA 336.14.3.0 but not in ICC 14216 K, irrespective of the fungal inoculum density, and overcame the resistance in UC 27 only at the higher inoculum density. Infection by the nematode significantly increased the number of propagules of F. oxysporum f. sp. ciceris race 5 in root tissues of genotypes with complete resistance to Fusarium wilt, compared with roots that were not inoculated with the nematode, irrespective of the fungal inoculum density, except in ICC 14216 K, in which this effect occurred only at the higher inoculum density. Reproduction of an M. artiellia population from Syria in the absence of F. oxysporum f. sp. ciceris race 5 was significantly higher than that of a population from Italy in all tested chick-pea genotypes except ICC 14216 K. However, there was no significant difference between the reproduction rates of the two nematode populations in plants infected with F. oxysporum f. sp. ciceris race 5, irrespective of the fungal inoculum density and the reaction of the genotypes to the fungus.     

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.     

Influence of Temperature and Inoculum Density of Fusarium oxysporum f. sp. ciceris on Suppression of Fusarium Wilt of Chickpea by Rhizosphere Bacteria

The effects of temperature and inoculum density of Fusarium oxysporum f. sp. ciceris race 5 on suppression of Fusarium wilt in chickpea (Cicer arietinum) cv. PV 61 by seed and soil treatments with rhizobacteria isolated from the chickpea rhizosphere were studied in a model system. Disease development over a range of temperatures (20, 25, and 30 degrees C) and inoculum densities (25 to 1,000 chlamydospores per gram of soil) was described by the Gompertz model. The Gompertz relative rate of disease progress and final amount of disease increased exponentially and monomolecularly, respectively, with increasing inoculum densities. Disease development was greater at 25 degrees C compared with 20 and 30 degrees C. At 20 and 30 degrees C, disease development was greater at 250 to 1,000 chlamydospores per gram of soil compared with 25 to 100 chlamydospores per gram of soil. At 25 degrees C, increasing inoculum densities of the pathogen did not influence disease. Nineteen Bacillus, Paenibacillus, Pseudomonas, and Stenotrophomonas spp. out of 23 bacterial isolates tested inhibited F. oxysporum f. sp. ciceris in vitro. Pseudomonas fluorescens RGAF 19 and RG 26, which did not inhibit the pathogen, showed the greatest Fusarium wilt suppression. Disease was suppressed only at 20 or 30 degrees C and at inoculum densities below 250 chlamydospores per gram of soil. Bacterial treatments increased the time to initial symptoms, reduced the Gompertz relative rate of disease progress, and reduced the overall amount of disease developed.     

Interactions between Fusarium oxysporum f. sp. tracheiphilum and Meloidogyne spp. in Vigna unguiculata. 1. Effects of different inoculum densities on Fusarium wilt

The effects of chlamydosporesandconidia of Fusarium oxysporum f sp. tracheiphilum at different initial spore concentrations were compared in the wilt-susceptible cowpea cultivar California Blackeye No. 5 (CB5). In glasshouse experiments with one inoculum density of either Meloidogyne incognita or M javanica, chlamydospores resulted in greater incidence and severity of Fusarium wilt than conidia at the same inoculum densities. Wilt symptoms also increased on wilt-resistant cultivar CB3 as inoculum densities of M. javanica were increased. When three cultivars were infested with moderate or high densities of both F. o. tracheiphilum and M. javanica. only CB5 developed sere wilt at either inoculum density. The wilt-tolerant cultivar Grant had mild wilt symptoms in most plants at moderate inoculum densities, and a tenfold increase in inoculum did not increase wilt ratings. CB3. however, had higher incidence and severity of Fusarium wilt symptoms at high inoculum densities, although 60% of the plants survived for 9 weeks.     

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.     

Interactions between Fusarium oxysporum f. sp. racheiphilum and Meloidogyne spp. in Vigna unguculata. 2. Specificity of different taxa

A California isolate of Meloidogyne javanica increased Fusarium wilt symptoms in cowpea cultivars California Blackeye No. 3 (CB3) (resistant to wilt) and Grant (tolerant) inoculated with each of the three races of Fusarium oxysporum f. sp. tracheiphilum. The same isolate of M. javancia did not similarly increase wilt in wilt-resistant cultivar CB7977 inoculated with two isolates of race 3 of F. o. tracheiphilum. Six of seven isolates of M. javanica caused similar increases in vascular discoloration in cultivar CB3. but one isolate of M. javanica and seven of M. incognita did not. Vascular discoloration rating was positively correlated with galling severity. However, increasing the initial inoculum density, and thus galling index, of one isolate of M. incognita did not increase vascular discoloration. The vascular discoloration ratings for the wilt-susceptible CB5 controls in each experiment were higher than those for the wilt-resistant cultivars infected with M. javanica. It is hypothesized that M. javanica but not M. incognita reduces, but does not eliminate, resistance to all races of F. o. tracheiphilum in cultivars CB3 and Grant.     

Development of a Specific Polymerase Chain Reaction-Based Assay for the Identification of Fusarium oxysporum f. sp. ciceris and Its Pathogenic Races 0, 1A, 5, and 6

ABSTRACT Specific primers and polymerase chain reaction (PCR) assays that identify Fusarium oxysporum f. sp. ciceris and each of the F. oxysporum f. sp. ciceris pathogenic races 0, 1A, 5, and 6 were developed. F. oxysporum f. sp. ciceris- and race-specific random amplified polymorphic DNA (RAPD) markers identified in a previous study were cloned and sequenced, and sequence characterized amplified region (SCAR) primers for specific PCR were developed. Each cloned RAPD marker was characterized by Southern hybridization analysis of Eco RI-digested genomic DNA of a subset of F. oxysporum f. sp. ciceris and nonpathogenic F. oxysporum isolates. All except two cloned RAPD markers consisted of DNA sequences that were found highly repetitive in the genome of all F. oxysporum f. sp. ciceris races. F. oxysporum f. sp. ciceris isolates representing eight reported races from a wide geographic range, nonpathogenic F. oxysporum isolates, isolates of F. oxysporum f. spp. lycopersici, melonis, niveum, phaseoli, and pisi, and isolates of 47 different Fusarium spp. were tested using the SCAR markers developed. The specific primer pairs amplified a single 1,503-bp product from all F. oxysporum f. sp. ciceris isolates; and single 900- and 1,000-bp products were selectively amplified from race 0 and race 6 isolates, respectively. The specificity of these amplifications was confirmed by hybridization analysis of the PCR products. A race 5-specific identification assay was developed using a touchdown-PCR procedure. A joint use of race 0- and race 6-specific SCAR primers in a single-PCR reaction together with a PCR assay using the race 6-specific primer pair correctly identified race 1A isolates for which no RAPD marker had been found previously. All the PCR assays described herein detected up to 0.1 ng of fungal genomic DNA. The specific SCAR primers and PCR assays developed in this study clearly identify and differentiate isolates of F. oxysporum f. sp. ciceris and of each of its pathogenic races 0, 1A, 5, and 6.     

Yield loss in chickpeas in relation to development of fusarium wilt epidemics

ABSTRACT Development of 108 epidemics of Fusarium wilt of chickpea caused by Fusarium oxysporum f. sp. ciceris were studied on cvs. P-2245 and PV-61 in field microplots artificially infested with races 0 and 5 of F. oxysporum f. sp. ciceris in 1986 to 1989. Disease progression data were fitted to the Richards model using nonlinear regression. The shape parameter was influenced primarily by date of sowing and, to a lesser extent, by chick-pea cultivars and races of F. oxysporum f. sp. ciceris. Fusarium wilt reduced chickpea yield by decreasing both seed yield and seed weight. These effects were related to sowing date, chickpea cultivar, and virulence of the prevalent F. oxysporum f. sp. ciceris race. Regression models were developed to relate chickpea yield to Fusarium wilt disease intensity with the following independent variables: time to initial symptoms (t(is)), time to inflection point (t(ip)) of the disease intensity index (DII) progress curve, final DII (DII(final)), standardized area under DII progress curve (SAUDPC), and the Richards weighted mean absolute rate of disease progression (rho). Irrespective of the chickpea cultivar x pathogen race combination, the absolute and relative seed yields decreased primarily by delayed sowing. The relative seed yield increased with the delay in t(is) and t(ip) and decreased with increasing DII(final), SAUDPC, and rho. A response surface as developed in which seed yield loss decreased in a linear relationship with the delay in t(is) and increased exponentially with the increase of rho.     

A Fusarium Wilt Resistance Gene in Gossypium barbadense and Its Effect on Root-Knot Nematode-Wilt Disease Complex

ABSTRACT Fusarium wilt, caused by the soilborne pathogen Fusarium oxysporum f. sp. vasinfectum race 1, is a vascular disease in cotton (Gossypium spp.), and is a component of a disease complex with root-knot nematodes (Meloidogyne incognita). Genetic analysis of two interspecific crosses (G. barbadense Pima S-7 x G. hirsutum Acala NemX and Pima S-7 x Acala SJ-2) showed that one major gene (designated Fov1) with allele dosage effect conferred resistance to F. oxysporum f. sp. vasinfectum race 1 in Pima S-7. Two amplified fragment length polymorphism (AFLP) markers were linked to Fov1 in Pima S-7, with genetic distance from the gene of 9.3 and 14.6 centimorgans. Less severe wilt symptoms in Acala NemX than Acala SJ-2 indicated that Acala NemX possesses one or more minor genes contributing to delay of wilt symptoms. Highly resistant plants in F(2) and F(3) (Pima S-7 x NemX) families indicated transgressive segregation effects of minor genes in Acala NemX combined with Fov1 from Pima S-7. The effects of wilt and nematode resistance on the nematode-wilt disease complex were assayed with two inoculation methods. In the presence of both pathogens, wilt damage measured as shoot and root weight reductions was greatest on wilt- and nematode-susceptible Acala SJ-2 and least in root-knot nematode-resistant and wilt-susceptible Acala NemX. Intermediate damage occurred in wilt-resistant and root-knot nematode-susceptible Pima S-7. The results indicated that nematode resistance was more effective than wilt resistance in suppressing wilt symptoms when either resistance was present alone. Nematode resistance combined with intermediate wilt resistance, as in the F(1) (Pima S-7 x NemX), was highly effective in protecting plants from root-knot nematodes and race 1 of Fusarium wilt as a disease complex.     

Association of induction of protease and chitinase in chickpea roots with resistance to Fusarium oxysporum f.sp. ciceri

To improve the breeding of chickpea varieties with resistance to Fusarium wilt, an attempt was made to analyse the biochemical basis of disease resistance by measuring levels of chitinase, β-1,3-glucanase, protease and proteinase inhibitor activities in dry and soaked seeds and in root and shoot tissues of wilt-resistant and wilt-susceptible cultivars. Marginal variation was observed in the levels of the candidate proteins in dry or soaked seeds. Chitinase activity was higher in roots than in shoots or cotyledons. No proteinase inhibitor activity was detected in root and shoot tissue of any of the cultivars. When the levels of these proteins were analysed in resistant (Vijay) and susceptible (JG-62) cultivars during development of wilt by growing plants in soil infested with F. oxysporum f.sp. ciceri , race 1, both cultivars showed induction of chitinase activity in the roots. However, the induced activity in JG-62 (3.82 U g⁻¹) was equivalent to the constitutive level in Vijay (3.90 U g⁻¹) and much lower than that induced in Vijay (5.18 U g⁻¹). Induction of protease activity was observed only in root extracts of Vijay when challenged by the pathogen. The root extract of Vijay showed in vitro antifungal activity in a plate assay. Simultaneous induction of proteolytic and chitinolytic activities specifically in the resistant cultivar was correlated with antifungal properties of root extracts effective in conferring resistance.     

Effect of root-knot nematodes on cajanol accumulation in the vascular tissues of pigeonpea after stem inoculation with Fusarium udum

The effect of root-knot nematodes ( Meloidogyne javanica and M. incognita ) on production of the isoflavonoid phytolexin, cajanol, was investigated in pigeonpea ( Cajanus cajan ) plants infected with Fusarium udum , the causal organism of Fusarium wilt. Seven-day-old seedlings of a wilt-resistant pigeonpea cultivar, ICP 9145 and a wilt-susceptible cultivar, Malawi Local, both of which were moderately susceptible to the nematode, were grown in soil infested with 2000 Meloidogyne juyeniles per plant. A duplicate set of plants remained free from nematodes. Twenty-one days later, all the plants were inoculated with F. udum by stem puncture. Quantitative estimates of cajanol in the vascular tissues were made at intervals up to 15 days after inoculation with the fungus. No external symptoms of wilt appeared in any plants of the wilt-resistant cultivar in the absence of the nematode. However, when inoculated with the nematode, two thirds of the plants developed wilt symptoms. Cajanol levels were lower in both the wilt-resistant and wilt-susceptible plants in the presence of the nematode than in its absence, although this effect was considerably more marked in the wilt-resistant cultivar. These results indicate that the root-knot nematode is capable of breaking resistance of ICP 9145 to Fusarium wilt and that at least part of the mechanism of this effect is retarded cajanol accumulation.     

Gene genealogies support Fusarium oxysporum f. sp. ciceris as a monophyletic group

Fusarium oxysporum f. sp. ciceris (Foc), the causal agent of fusarium wilt of chickpea, consists of two pathotypes (yellowing and wilting) and eight races (races 0, 1B/C, 1A and 2–6) of diverse geographical distribution. Six Foc isolates, one each of races 0, 1B/C, 1A, 4, 5 and 6, representing the two pathotypes and the geographical range of the pathogen, showed identical sequences in introns of the genes for translation elongation factor 1α ( EF1 α), β-tubulin, histone 3, actin and calmodulin. Eleven additional Foc isolates representative of all races, pathotypes and geographical range, and three isolates of F. oxysporum (Fo) nonpathogenic to chickpea were further analysed for sequence variation in the EF1 α gene. All isolates pathogenic to chickpeas shared an identical EF1 α gene sequence, which differed from that shared by the three Fo isolates nonpathogenic to chickpea. EF1 α gene sequences from the 17 Foc isolates and the three Fo isolates were compared with 24 EF1 α gene sequences in GenBank from isolates of 11 formae speciales of F. oxysporum by parsimony analysis. Foc isolates formed a grouping distinct from other formae speciales and nonpathogenic isolates. These results indicate that F. oxysporum f. sp. ciceris is monophyletic.     

Influence of chickpea genotype and Bacillus sp. on protection from Fusarium wilt by seed treatment with nonpathogenic Fusarium oxysporum

Seeds of kabuli chickpea cultivars ICCV 4 and PV 61 were treated with conidia of nonpathogenic Fusarium oxysporum isolate Fo 90105 suspended in methylcellulose (3 × 10⁶ conidia.seed^-1), or with methylcellulose alone, and sown in soil artificially infested with 500 or 1,000 chlamydospores.g^-1 of F. oxysporum f. sp. ciceris race 5. At an inoculum concentration of 500 chlamydospores.g^-1, seed treatment with Fo 90105 significantly increased the incubation period of the disease by 11 (ICCV 4) or 25 (PV 61) days, and reduced the final disease incidence, disease intensity and the standardized area under the curve of disease intensity over time. This protection from disease was higher and more consistent in PV 61 than in ICCV 4. However, it was annulled with an inoculum concentration of 1,000 chlamydospores.g^-1, except for the incubation period in PV 61 which was increased by 10 days. When ICCV 4 seeds were treated with Fo 90105 (3 × 10⁶ conidia.seed^-1) and/or Bacillus sp. isolate RGAF 51 (1 × 10⁷ cfu.seed^-1), then sown in infested soil, there was no influence by the Bacillus isolate on protection conferred by Fo 90105. However, the degree of protection by the nonpathogenic F. oxysporum was higher and more consistent when plants from treated seeds were grown in sterile sand for 6 days, then transplanted into infested soil.     

Interactions between Fusarium oxysporum f. sp. tracheiphilum and Meloidogyne spp. in Vigna unguiculata. 3. Pathogenesis by F. o. tracheiphilum as affected by M. javanica and host cultivar

Experiments were conducted to determine the temporal and spatial effects of Meloidogyne jav anica and host cultivar on pathogenesis by Fusarium oxysporum f. sp tracheiphilum in cowpea. In the wilt-susceptible cowpea cultivar California Blackeye No. 5 (CB5). F. o. tracheiphilum proliferated rapidly in both the hypocotyl and first internode 6 weeks after inoculation. The fungus spread quickly upward as plants grew, colonized most tissues within 6 weeks, and caused severe wilt. In wilt-resistant cultivar CB3. there was little proliferation of F. oxysporum in any tissue, whether or not plants were infected by St. javanica. The fungus was found above the primary internode in 15"., of all CB3 plants, but did not continue to spread upward after 4 weeks Vascular discoloration was greatest when St. javanica was added 4 weeks before F. o. tracheiphilum, but simultaneous inoculation also increased wilt symptoms Root wounding did not increase wilt. Split-root experiments provided no evidence that infection by, M. javanica results in a translocatable factor that reduces wilt resistance. When hypocoivls were inoculated with F. o. tracheiphilum at different intervals after roots infected by St. jav anica were removed, there was no evidence that the effect of the nematode on wilt susceptibility was translocated or persistent.     

Root exudates associated with the resistance of four chickpea cultivars (Cicer arietinum) to two races of Fusarium oxysporum f.sp. ciceri

The germination of race 1 spores of Fusarium oxysporum f. sp. ciceri was significantly inhibited by the root exudate of the wilt-resistant chickpea cvs CPS1 and WR315 compared to untreated spores and spores treated with root exudates from susceptible cultivars. The effect was concentration dependent, such that the exudate from 1 g of root in 2 ml of water almost completely inhibited spore germination, whereas the exudate from 1 g of root in 20 ml water did not do so. The inhibitory effects of the active exudates were negated when the apolar components of the exudates were removed by extraction with ethyl acetate. The root exudates of the susceptible cv. JG62 and the late wilting cv. H208 did not inhibit germination. The hyphal growth of germinated spores was also strongly inhibited by the concentrated exudates of CPS1 and WR315, and diluted exudates were less potent. The highest concentration of the exudate of the susceptible cv. JG62 showed some inhibition of hyphal growth, whereas none of the exudates of H208 were found to contain any antifungal activity. The effect of the exudates on the spores of race 2 was similar to that reported for race 1, except that the water-soluble components of the crude root exudate of WR315 after ethyl acetate extraction were also found to inhibit germination significantly. Overall, the spores of race 2 appeared to be more susceptible to the effects of the exudates. The ethyl acetate fractions of the root exudates of CPS1 and WR315 strongly inhibited germination and hyphal growth of both race 1 and race 2, the effect being concentration dependent. The results suggest that the resistance of chickpeas to vascular wilt depends, at least in part, upon the antifungal activity of the root exudates. Differences in the expression of resistance in the field could depend upon the concentration or rate of production of constitutive antifungal components by the root.     

Parasitism of the root-lesion nematode Pratylenchus thornei on chickpea

Pratylenchus thornei -chickpea interactions were investigated under controlled and fluctuating environmental conditions in the growth chamber, greenhouse and shadehouse. Under controlled conditions, P. thornei infected chickpea lines 12071/10054 and P2245 and cultivars Andoum 1, JG62 and UC27. Line P 2245 and cv. JG 62 were the most susceptible genotypes on the basis of root damage and nematode reproduction, but nematode infection did not significantly reduce root and shoot weights. Cultivars Andoum 1 and UC27 and line 12071/10054 showed the least root damage and nematode reproduction. Inoculation of cv. Andoum 1 with 2500, 5000 or 10000 nematodes per plant in pots did not affect shoot weight, regardless of the conditions of water stress of the plants. However, root weight was significantly reduced by nematode infection in plants grown under water stress and fluctuating temperature conditions in the greenhouse, but was not affected by any other treatment. The nematode reproduction index was not affected by soil water content under shadehouse conditions, but was greater on plants watered to soil water-holding capacity than in water-stressed plants under greenhouse conditions. For both environments, the nematode reproduction index decreased when inoculum density was greater than 5000 nematodes per plant.     

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.     

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.     

Quantitative Modeling of the Effects of Temperature and Inoculum Density of Fusarium oxysporum f. sp. ciceris Races 0 and 5 on Development of Fusarium Wilt in Chickpea Cultivars

ABSTRACT Races 0 (Foc-0) and 5 (Foc-5) of Fusarium oxysporum f. sp. ciceris differ in virulence and induce yellowing or wilting syndrome, respectively, in chickpea. We modeled the combined effects of soil temperature and inoculum density of Foc-0 and Foc-5 on disease developed in chickpea cvs. P-2245 and PV-61 differing in susceptibility to those races, using quantitative nonlinear models. Disease development over time in the temperature range of 10 to 30 degrees C and inoculum densities between 6 and 8,000 chlamydospores g(1) of soil was described by the Weibull function. Four response variables (the reciprocal incubation period, the final disease intensity, the standardized area under the disease progress curve, and the intrinsic rate of disease development) characterized the disease development. Response surface models that expressed the combined effect of inoculum density and temperature were developed by substituting the intrinsic rate of disease development in the Weibull or exponential functions with a beta function describing the relationship of response variables to temperature. The models estimated 22 to 26 degrees C as the most favorable soil temperature for infection of cvs. P-2245 and PV-61 by Foc-5, and 24 to 28 degrees C for infection of cv. P-2245 by Foc-0. At 10 degrees C, no disease developed except in cv. P-2245 inoculated with Foc-5. At optimum soil temperature, maximum disease intensity developed with Foc-5 and Foc-0 at 6 and 50 chlamydospores g(1) of soil respectively, in cv. P-2245, and with Foc-5 at 1,000 chlamydospores g(1) of soil in cv. PV-61. The models were used to construct risk threshold charts that can be used to estimate the potential risk of Fusarium wilt epidemics in a geographical area based on soil temperature, the race and inoculum density in soil, and the level of susceptibility of the chickpea cultivar.     

United States Department of Agriculture-Agricultural Research Service studies on polyketide toxins of Fusarium oxysporum f sp vasinfectum: potential targets for disease control

A group of 133 isolates of the cotton wilt pathogen Fusarium oxysporum Schlecht f sp vasinfectum (Atk) Sny & Hans, representing five races and 20 vegetative compatibility groups within race 1 were used to determine the identity, biosynthetic regulation and taxonomic distribution of polyketide toxins produced by this pathogen. All isolates of F oxysporum f sp vasinfectum produced and secreted the nonaketide naphthazarin quinones, bikaverin and norbikaverin. Most isolates of race 1 (previously denoted as races 1, 2 and 6; and also called race A) also synthesized the heptaketide naphthoquinones, nectriafurone, anhydrofusarubin lactol and 5-O-methyljavanicin. Nine avirulent isolates of F oxysporum from Upland cotton roots, three isolates of race 3 of F oxysporum f sp vasinfectum, and four isolates of F oxysporum f sp vasinfectum from Australia, all of which previously failed to cause disease of Upland cotton (Gossypium hirsutum L) in stem-puncture assays, also failed to synthesize or secrete more than trace amounts of the heptaketide compounds. These results indicate that the heptaketides may have a unique role in the virulence of race 1 to Upland cotton. The synthesis of all polyketide toxins by ATCC isolate 24908 of F oxysporum f sp vasinfectum was regulated by pH, carbon/nitrogen ratios, and availability of calcium in media. Synthesis was greatest below pH 7.0 and increased progressively as carbon/nitrogen ratios were increased by decreasing the amounts of nitrogen added to media. The nonaketides were the major polyketides accumulated in synthetic media at pH 4.5 and below, whereas the heptaketides were predominant at pH 5.0 and above. The heptaketides were the major polyketides formed when 10 F oxysporum f sp vasinfectum race 1 isolates were grown on sterilized stems of Fusarium wilt-susceptible cotton cultivars, but these compounds were not produced on sorghum grain cultures. Both groups of polyketide toxins were apparently secreted by F oxysporum f sp vasinfectum, since half of the toxin in 2-day-old shake culture was present in the supernatant. Secretion was enhanced by calcium. Glutamine and glutamic acid inhibited both nonaketide and heptaketide syntheses, even at low nitrogen