Reduction of Fusarium rot and maintenance of fruit quality in melon using eco-friendly hot water treatment

Significant losses in harvested fruit can be directly attributable to decay fungi and quality deterioration. Hot water treatment (HWT) has been demonstrated to be an effective and economic environment-friendly approach for managing postharvest decay and maintaining fruit quality. In this study, the effects of HWT (45 °C for 10, 15, 20, and 25 min) on in vitro growth of Fusarium oxysporum, in vivo Fusarium rot, and natural decay of melon were investigated. HWT inhibited spore germination and germ tube elongation of F. oxysporum. Protein impairment and ATP consumption triggered by HWT contributed to the inhibitory effect. Results of in vivo studies showed that HWT effectively controlled Fusarium rot and natural decay of melon. Correspondingly, HWT induced a significant increase in content of total phenolic compounds and lignin of melon. These findings indicate that the effects of HWT on Fusarium rot may be associated with the direct fungal inhibition and the elicitation of defense responses in fruit. Importantly, HWT used in this study had beneficial effects on fruit quality as well. HWT may represent an effective non-chemical approach for management of postharvest Fusarium rot.     

Comparison of in vitro activities of amphotericin, clotrimazole, econazole, miconazole, and nystatin against Fusarium oxysporum.

The inhibitory activity of amphotericin B, clotrimazole, econazole, miconazole and nystatin was compared against Fusarium oxysporum f.sp. radicis-cucumerinum. The most efficient antifungal agent against the growth of Fusarium oxysporum was econazole, followed by clotrimazole, miconazole, amphotericin and nystatin. The ED50 and ED90 values were 0.053 and 1.002 ppm for econazole, 0.088 and 1.100 ppm for clotrimazole, 0.173 and 3.210 ppm for miconazole, 0.713 and greater than 48 ppm for amphotericin and 3.860 and 16.702 ppm for nystatin. The ED50 values of nystatin and amphotericin against spore germination of Fusarium oxysporum were determined at 3.1427 ppm and 8.3990 ppm respectively, nystatin was 2.76 times more effective than amphotericin, while no effect was observed after the addition of econazole, clotrimazole and miconazole. The tested azoles were more effective than amphotericin and nystatin on growth inhibition of Fusarium oxysporum but amphotericin and nystatin acted significantly better on spore germination of Fusarium.     

Efficacy of sludge and manure compost amendments against Fusarium wilt of cucumber

Fusarium wilt of cucumber caused by the fungus, Fusarium oxysporum, is one of the most destructive soilborne diseases and can result in serious economic loss. No efficient fungicide is currently available to control the disease. The aim of this study was to examine the disease suppression ability of pig manure and sludge composts in peat-based container media and explore the possible disease suppression mechanisms. Pig manure and sewage sludge compost were made in laboratory-scale tanks. Plant growth media were formulated with peat mixture and compost (or 60?°C heated compost) in a 4:1 ratio (v/v). Cucumber seedlings were artificially inoculated with F. oxysporum conidia (5?×?10⁵ conidia mL^?1) by the root-dip method. Cucumber Fusarium wilt was effectively suppressed in sludge compost-amended media, while the disease suppression effect of pig manure compost was limited. The ammonia levels in the manure compost-amended media were significantly higher than those of sludge compost-amended media, which could explain its lower disease suppression ability. Heated composts behaved similarly with respect to disease suppression. Adding composts increased microbial biomass, microbial activity, and the microbial diversity of the growth media. PCR-DGGE results indicated that the fungal community had a significant correlation to the disease severity. The artificially inoculated pathogen was retrieved in all treatments and one possible biocontrol agent was identified as a strain of F. oxysporum by phylogenetic analyses. The results indicated that the sludge compost used in this study could be applied as a method for biocontrol of cucumber Fusarium wilt.     

Extraction and purification of DNA from phytopathogenic soil-inhabiting Fusarium species

Working quantities of good quality DNA was obtained from three pathogenic forms of the soil-inhabiting fungus, Fusarium oxysporum (FO), which causes economically significant diseases in tomato crops. The mitochondrial, nuclear and ribosomal fractions from each of the DNAs were separated by cesium chloride gradient centrifugations. The data suggest a decrease in nuclear DNA and an increase in mitochondrial DNA correlated with increasing pathogenicity among the three pathogens examined. Ribosomal DNA was detected only in FO, the least pathogenic form.     

Halofenate and clofibrate inhibition of pyruvate dehydrogenase from Fusarium culmorum

Pyruvate dehydrogenase (E1, E.C. 1.2.4.1) was obtained from Fusarium culmorum by ammonium sulfate precipitation. An eight-fold purification was obtained with a specific activity of 13 K units/mg protein. Both halofenate and clofibrate inhibited the enzyme complex non-competitively. The inhibitory effect of halofenate was greater than that of clofibrate being 42% higher at 20 mM concentration compared to the inhibition by clofibrate at 40 mM concentration. Both compounds disorganized the normal cytoplasmic lipids including the emptying of cells in the mycelium suggesting membrane disruption.     

PCR-RAPD profiling of Fusarium spp. causing guava wilt disease in India

Wilt is a serious disease of the guava crop in India. Fusarium oxysporum f. sp. psidii and F. solani have been reported as causative agents of this disease. In this study, 42 isolates each of F. oxysporum f. sp. psidii and F. solani, were isolated from guava cultivars and characterized by randomly amplified polymorphic DNA (RAPD) method. Thirty RAPD primers were tested in the genome of Fusarium spp. and the number of scorable bands for corresponding primer ranged from 1-8 with an average of 5 bands per individual. DNA band size ranged from 200 bp to 5090 bp. A 0.21 per cent polymorphism was found in individual isolates of F. solani indicating that the 42 isolates were similar. However, a 2.58 percent polymorphism among individual isolates of F. oxysporum f.sp. psidii showed a higher level of genetic diversity. Cluster analysis of the RAPD band patterns clearly separated the isolates of F. oxysporum f.sp. psidii into three clusters. Two clusters were formed with F. solani isolates, showing a higher degree of similarity. Unique fingerprint profiles generated by the PCR-RAPD can be exploited for genetic characterization purposes.     

Antimicrobial effects of clofibrate on the wheat pathogen Fusarium culmorum

Abstract

The biological effects of clofibrate (ethyl p‐chlorophenoxy‐isobutyric acid) on the growth and metabolism of the soil‐borne wheat pathogen Fusarium culmorum, were examined.

In mid log phase (16 hr) cultures both phenylalanine uptake and secondary spore production were stimulated at 0.1 μM concentration; the net sterol content was reduced 50% at 0.35 μM; oxygen uptake was stimulated at 0.1 mM; growth was inhibited 50% at 0.1 mM concentration. Both phenylalanine and oxygen uptake were inhibited at 1.0 mM and pyruvate dehydrogenase activity was reduced 50% at 50 mM concentration of clofibrate.

The data indicate that clofibrate affects a number of biological and enzyme systems. The inhibitory effect on the growth of the pathogen suggest a potential use of hypolipidemic agents like clofibrate as an antifungal agent for seed protection.     

Polycyclic aromatic hydrocarbons storage by Fusarium solani in intracellular lipid vesicles

Accumulation and elimination of polycyclic aromatic hydrocarbons (PAHs) were studied in the fungus Fusarium solani. When the fungus was grown on a synthetic medium containing benzo[a]pyrene, hyphae of F. solani contained numerous lipid vesicles which could be stained by the lipid-specific dyes: Sudan III and Rhodamine B. The fluorescence produced by Rhodamine B and PAH benzo[a]pyrene were at the same locations in the fungal hyphae, indicating that F. solani stored PAH in pre-existing lipid vesicles. A passive temperature-independent process is involved in the benzo[a]pyrene uptake and storage. Sodium azide, a cytochrome c oxidation inhibitor, and the two cytoskeleton inhibitors colchicine and cytochalasin did not prevent the transport and accumulation of PAH in lipid vesicles of F. solani hyphae. F. solani degraded a large range of PAHs at different rates. PAH intracellular storage in lipid vesicles was not necessarily accompanied by degradation and was common to numerous other fungi.     

Microbial degradation of trifluralin by Aspergillus carneus, Fusarium oxysporum and Trichoderma viride

3H-Trifluralin was synthesized by condensation of 3H-4-chloro-3,5-dinitro-alpha, alpha, alpha-trifluorotoluene with di-n-propylamine. After incubation of trifluralin with Aspergillus carneus, Fusarium oxysporum and Trichoderma viride for 10 days, a small percentage (less than 10%) of unchanged herbicide was recovered in the extractable fraction. This indicates a fairly rapid degradation of the herbicide by the fungal species. Other than trifluralin, the culture medium contained at least five labelled products: 2,6-dinitro-N-n-propyl-alpha, alpha, alpha-trifluoro-p-toluidine; 2,6-dinitro-alpha, alpha, alpha-trifluoro-p-toluidine; 2-amino-6-nitro-alpha, alpha, alpha-trifluoro-p-toluidine, 2,6-dinitro-4-trifluoromethyl phenol and a major polar product which constituted more than 50% of the total extractable transformation products. A pathway, which simulates that of aerobic degradation of the herbicide in soil, is suggested for the microbiological degradation of trifluralin.     

Bioconversion of olive-mill dry residue by Fusarium lateritium and subsequent impact on its phytotoxicity

The present study investigated the ability of the non-pathogenic fungus Fusarium lateritium to either degrade or modify aromatic substances in olive-mill dry residue (DOR) and to reduce its phytotoxicity. The 80% reduction of ethylacetate extractable phenols in DOR colonized by the fungus for 20 weeks appeared to be due to polymerization reactions of phenol molecules as suggested by mass-balance ultrafiltration and size-exclusion chromatography experiments. Several lignin-modifying oxidases, including laccase, Mn-peroxidase and Mn-inhibited peroxidase were detected in F. lateritium solid-state cultures. Tests performed with tomato seedlings in soils containing 6% (w/w) sterilized non-inoculated DOR showed that the waste was highly phytotoxic. By contract, F. lateritium growth on DOR for 20 weeks led to a complete removal of the waste toxicity and to a higher shoot dry weight of tomato plants than that obtained in the absence of DOR.     

Effect of common food preservatives on mycelial growth and spore germination of Fusarium oxysporum

The growth and spore germination inhibition of Fusarium oxysporum f.sp. radicis-cucumerinum by the common food additives: acetic acid, formic acid potassium sorbate, propionic acid, sorbic acid, and the fungistatic agent sec-butylamine was examined in vitro. The inhibitory efficacy of these chemicals decreased in the following order: sorbic acid, potassium sorbate, propionic acid, acetic acid, sec-butylamine and formic acid. At pH 6.4, the ED50 value for mycelium growth was: 976 ppm for sorbic acid, 1292 ppm for potassium sorbate, 2435 ppm for propionic acid, 3805 ppm for acetic acid, 3962 ppm for sec butylamine and 4668 ppm for formic acid. The ED50 value for spore germination was: 225 ppm for potassium sorbate, 1201 ppm for sorbic acid, 1402 ppm for propionic acid, 1600 ppm for sec-butylamine, 1957 ppm for acetic acid and 2485 ppm for formic acid.     

Effect of common food preservatives on mycelial growth and spore germination of Fusarium oxysporum

Abstract

The growth and spore germination inhibition of Fusarium oxysporum f.sp. radicis‐cucumerinum by the common food additives: acetic acid, formic acid potassium sorbate, propionic acid, sorbic acid, and the fungistatic agent sec‐butylamine was examined in vitro. The inhibitory efficacy of these chemicals decreased in the following order: sorbic acid, potassium sorbate, propionic acid, acetic acid, sec‐butylamine and formic acid. At pH 6.4, the ED₅₀ value for mycelium growth was: 976 ppm for sorbic acid, 1292 ppm for potassium sorbate, 2435 ppm for propionic acid, 3805 ppm for acetic acid, 3962 ppm for sec butylamine and 4668 ppm for formic acid. The ED₅₀ value for spore germination was: 225 ppm for potassium sorbate, 1201 ppm for sorbic acid, 1402 ppm for propionic acid, 1600 ppm for sec‐butylamine, 1957 ppm for acetic acid and 2485 ppm for formic acid.     

Potentiality of different isolates of wilt fungus Fusarium oxysporum collected from rhizosphere of tomato against root-knot nematode Meloidogyne incognita

This investigation was undertaken to determine the effect of culture filtrates of different isolates of Fusarium oxysporum f. sp. lycopersici on mortality of Meloidogyne incognita juveniles and egg hatching. It was observed that different concentrations including standard extract (S.E), 1:10 and 1:100 dilutions of all fungal filtrates inhibited egg hatch when compared with control. Minimum mortality and maximum hatching was observed in BRT (showing least mortality) isolate of F. oxysporum, while maximum mortality and minimum egg hatching was recorded in BGT (showing maximum mortality) isolate. Larval mortality was decreased with a decrease in concentration and the least mortality was observed in 1:100 when compared with SE and 1:10. The potentiality of both the isolates (BRT and BGT) against root-knot nematode M. incognita was confirmed by the pathogenicity test on tomato. These observations confirmed that F. oxysporumisolates possesses variability in pathogenicity ranging from pathogenic to bio-control agent. The plants inoculated with BRT isolate failed to show wilt symptoms while plants inoculated with BGT isolate showed wilt indices.     

Antibiotic effect of exogenously applied salicylic acid on in vitro soilborne pathogen, Fusarium oxysporum f.sp.niveum

Salicylic acid, which is biosynthesized inside plant and is often found and accumulated in soil due to plant debris decaying, is considered as a signaling substance during plant-microbe interactions. It is involved in the cycling of biogeochemistry and related to plant resistance to biotic and abiotic stress. The antibiotic effect of salicylic acid on Fusarium oxysporum f.sp.niveum (FON) was studied to investigate the relationships between the salicylic acid and the fungus in the ecological interaction of plant-microbe. Results showed that the biomass, colony diameter, number of conidium germination and conidium production of FON were decreased by 52.0%, 25.7%, 100% and 100% at concentrations of 800 mg L(-1). However, mycotoxin yield was increased by 233%, pectinase activity raised by 168.0% and cellulase activity increased by 1325% compared to control at higher concentrations. It was concluded that salicylic acid as an allelochemical greatly inhibited FON growth and conidia formation and germination, though stimulated mycotoxin production and activities of hydrolytic enzymes by FON.     

Growth and disease response of soybeans from early maturity groups to ozone and Fusarium oxysporum

The single and combined effects of ozone (O(3)) and Fusarium oxysporum on growth and disease expression of soybean genotypes differing in foliar sensitivity to O(3) were studied in the greenhouse. O(3) had no effect on root and hypocotyl rot severity of PI 153.283 (O(3)-sensitive, S) or PI 189.907 (O(3)-tolerant, T) maturity group I soybean lines. Plants of both genotypes infected with F. oxysporum and exposed to O(3) had greater reductions in relative growth rate (RGR), net assimilation rate (NAR), and had more stippled leaves per plant than Fusarium-free plants exposed to O(3). O(3) alone had a greater impact on shoot dry weight, RGR, and NAR of PI 153.283 (S) than of PI 189.907 (T). O(3) alone reduced shoot and root dry weights primarily through a depression in NAR and less through reduced leaf area. F. oxysporum alone reduced root dry weight at 35 days; however, infected plants responded with increases in root dry weight from 49 to 63 days. Similarly, F. oxysporum alone lowered early RGR but subsequent RGR decline was less rapid while NAR remained high, particularly during later sampling intervals. Infection by F. oxysporum that causes root and hypocotyl rot increased soybean sensitivity to O(3) by prolonging active vegetative growth.     

Interaction of ozone exposure and Fusarium subglutinans inoculation on growth and disease development of loblolly pine seedlings

Seedlings from ten half-sib families of loblolly pine (Pinus taeda) were exposed in open-top chambers to carbon-filtered air (CF), non-filtered air (NF), or air amended with ozone to 1.7 or 2.5 times ambient. After 105 days of exposure, half the seedlings within each family were wounded but not inoculated and half were wounded and inoculated with the pitch canker fungus, Fusarium subglutinans, to which five families were relatively resistant. After an additional 50 days of ozone treatment, seedling growth and canker development were recorded. Cankers were significantly (sigma < or = 0.05) smaller among resistant compared to susceptible families, and were significantly larger among seedlings receiving the highest (2.5) compared to the ambient (NF) ozone treatment. The wound scars of non-inoculated seedlings were also significantly larger among seedlings receiving the 2.5 compared to the NF treatment, but these dimensions did not differ significantly with seedling family or resistance. The weights of needles and large roots were significantly smaller at the 2.5 compared to the 1.7 ozone treatment for inoculated but not for non-inoculated seedlings; this resulted in a significant interaction for ozone and inoculation effects. Among resistant families, root weights were significantly smaller for inoculated seedlings. Diameter growth and dry weights of needles were significantly smaller among inoculated compared to non-inoculated seedlings, but did not differ between NF and 2.5 ozone treatments.     

Inhibition of the antioxidant activity of catalase and superoxide dismutase from Fusarium verticillioides exposed to a Jacquinia macrocarpa antifungal fraction

The aim of this study was to investigate the in vitro effect of an antifungal fraction obtained from Jacquinia macrocarpa plant (JmAF) in the generation of reactive oxygen species (ROS) and the activity of the catalase (CAT) and superoxide dismutase (SOD) enzymes from Fusarium verticillioides, as well as their influence in the viability of the fungus spores. The compounds present in the JmAF were determined by gas chromatography/quadrupole time-of-flight mass spectrometry (GC/QTOF-MS). The effect of the exposition to JmAF on the generation of ROS, as well as in the CAT and SOD activities in F. verticillioides, was determined. The main compounds detected were γ-sitosterol, stephamiersine, betulinol and oleic acid. JmAF showed very high ability in inhibiting the spore viability of F. verticillioides, and their capacity to cause oxidative stress by induction of ROS production. JmAF induced the highest ROS concentration and also inhibited CAT and SOD activities. The results obtained in this study indicate that JmAF is worthy of being considered for the fight against phytopathogenic fungi.     

Potentiality of different isolates of wilt fungus Fusarium oxysporum collected from rhizosphere of tomato against root-knot nematode Meloidogyne incognita

This investigation was undertaken to determine the effect of culture filtrates of different isolates of Fusarium oxysporum f. sp. lycopersici on mortality of Meloidogyne incognita juveniles and egg hatching. It was observed that different concentrations including standard extract (S.E), 1:10 and 1:100 dilutions of all fungal filtrates inhibited egg hatch when compared with control. Minimum mortality and maximum hatching was observed in BRT (showing least mortality) isolate of F. oxysporum, while maximum mortality and minimum egg hatching was recorded in BGT (showing maximum mortality) isolate. Larval mortality was decreased with a decrease in concentration and the least mortality was observed in 1:100 when compared with SE and 1:10. The potentiality of both the isolates (BRT and BGT) against root-knot nematode M. incognita was confirmed by the pathogenicity test on tomato. These observations confirmed that F. oxysporumisolates possesses variability in pathogenicity ranging from pathogenic to bio-control agent. The plants inoculated with BRT isolate failed to show wilt symptoms while plants inoculated with BGT isolate showed wilt indices.     

Preliminary evidence of the role of hydrogen peroxide in the degradation of benzo[a]pyrene by a non-white rot fungus Fusarium solani

In order to study the enzymatic mechanisms involved in the successive steps of BaP degradation by a Deuteromycete fungus Fusarium solani, we developed an indirect approach by using inhibitors of enzymes. We used either specific inhibitors of peroxidases (i.e. salicylhydroxamic acid) and of cytochrome P-450 (i.e. piperonyl butoxyde) or inhibitors of both enzymes (i.e. potassium cyanide). Surprisingly, no expected decrease of BaP degradation was observed with most inhibitors tested. On the contrary, more BaP was degraded. Only butylated hydroxytoluene, which acts as a free radical scavenger, inhibited BaP degradation. The inhibition of these enzymes, which use H(2)O(2) as a cosubstrate, might have resulted in an increase of hydrogen peroxide availability in the fungal cultures. This enhancement could induce formation of reactive oxygen species (ROS) which might be the agents that initiate benzo[a]pyrene oxidation. This study proposed a hypothetic alternative metabolic pathway involved in PAH metabolism by Fusarium solani.     

Comparison of antibiotic tolerance,lipids and respiration in the tomato pathogens Fusarium oxysporum lycopersici and F. oxysporum radicis lycopersici

Abstract

The respiration and lipid contents and the tolerance to mycostatin, chloramphenicol and cycloheximide were compared in the two morphologically similar forms of the tomato pathogens: Fusarium oxysporum lycopersici (FOL) and the virulent form F. oxysporum radicis lycopersici (FORL). The differential tolerances to mycostatin were the most significant feature of the comparisons. The MIC for FORL was 24 μg/mL for the mycelium and 38 μg/mL for the spores. For FOL, the MIC was 8 ug/mL for both. This pattern of higher mycostatin tolerance by FORL obtained at 19°C and 27°C. There were differences between FOL and FORL in their fatty acid composition. FORL contained about three times as much C_18:0 and over twice as much C_18:1 as FOL. Conversely FOL contains over two times as much C_16:1 as FORL. There appeared to be no significant differences between the respiration rates of the two pathogens. The data is discussed relative to their significance as the biochemical basis for examining pathogenicity and virulence between the two organisms.