Oxalic acid production by Fusarium oxysporum Schlecht and Botryodiplodia theobromae Pat., post-harvest fungal pathogens of yams (Dioscorea rotundata L.) and detoxification by Bacillus subtilis CM1 isolated from culturable cowdung microflora

Abstract

Fusarium oxysporum Schlecht and Botryodiplodia theobromae Pat., two important post-harvest pathogens of yam (Dioscorea rotundata L.) tubers in storage were found to produce oxalic acid (OA) in vitro and in vivo. The rate of OA accumulation was proportional to fungal growth (cell mass) in Potato Dextrose liquid medium during 10 days incubation period. Further, simultaneous co-culturing of either of the fungi with Bacillus subtilis CM1 isolated from cowdung culturable microflora resulted in 92% reduction in OA accumulation compared with that in the culture of the individual fungus. The effect was more prominent in pH 5 – 6 than in pH 7 – 8. B. subtilis CM1 was capable of detoxifying OA and several proteins were detected in the culture filtrates when it was grown in peptone-mineral salt medium containing OA. SDS-PAGE analysis of 70% ammonium sulphate fraction of the culture filtrate exhibited the presence of a predominant 97 kDa protein.     

Biological control of postharvest fungal rot of yam (<Emphasis Type="Italic">Dioscorea</Emphasis> spp.) with<Emphasis Type="Italic">Bacillus subtilis</Emphasis>

The potential of isolates of Bacillus subtilis from yam farm soil to control rot of yam in storage barns was investigated. Yam tubers inoculated in vivo with B. subtilis showed no rot while those inoculated with Aspergillus niger, Botryodiploidia theobromae or Penicillium oxalicum showed considerable rot. The set of yams in which B. subtilis and the fungi were simultaneously inoculated produced rot whereas those in which B. subtilis was inoculated a day before the fungi was inoculated were totally reduced or free of rot. Many fewer fungi were isolated from the surface of tubers treated with B. subtilis than from the untreated (control) and there was high recovery of B. subtilis (99–100%) throughout the period of storage. Rot build up was faster in uninoculated control tubers or those inoculated with a spoilage fungus, while those treated with the antagonist were totally reduced or free of rot. The culture filtrate of B. subtilis prevented spore germination in some spoilage fungi. The importance of this study in relation to farmers in developing countries is discussed.     

Impact of Bacillus subtilis JA, a biocontrol strain of fungal plant pathogens, on arbuscular mycorrhiza formation in Zea mays

Summary  In the present study, the influence of Bacillus subtilis JA on arbuscular mycorrhizal fungi (AMF) was evaluated by either pot culture or in vitro conditions, respectively. Under the pot culture conditions, the inoculation of B. subtilis JA decreased the frequency (% F) of the root colonization by indigenous arbuscular mycorrhizal fungi and the shoot dry weight of maize (Zea mays L.), but had no apparent effect on the intensity (% I) of AM fungal root colonization. The unknown volatile emitted from the B. subtilis JA in vitro significantly inhibited spore germination and the hyphal growth in the dual-compartment experiments. Moreover, the data from the direct interaction between B. subtilis JA and Glomus etunicatum showed that soluble antifungal lipopeptides influenced the development of AMF. Therefore, the application of antifungal Bacillus strains should take the compatibility with the indigenous beneficial fungi into consideration.     

Microbial conversion and in vitro and in vivo antifungal assessment of bioconverted docosahexaenoic acid (bDHA) used against agricultural plant pathogenic fungi

Microbial modification of polyunsaturated fatty acids can often lead to special changes in their structure and in biological potential. Therefore, the aim of this study was to develop potential antifungal agents through the microbial conversion of docosahexaenoic acid (DHA). Bioconverted oil extract of docosahexaenoic acid (bDHA), obtained from the microbial conversion of docosahexaenoic acid (DHA) by Pseudomonas aeruginosa PR3, was assessed for its in vitro and in vivo antifungal potential. Mycelial growth inhibition of test plant pathogens, such as Botrytis cinerea, Colletotrichum capsici, Fusarium oxysporum, Fusarium solani, Phytophthora capsici, Rhizoctonia solani and Sclerotinia sclerotiorum, was measured in vitro. bDHA (5 μl disc⁻¹) inhibited 55.30–65.90% fungal mycelium radial growth of all the tested plant pathogens. Minimum inhibitory concentrations (MICs) of bDHA against the tested plant pathogens were found in the range of 125–500 μg ml⁻¹. Also, bDHA had a strong detrimental effect on spore germination for all the tested plant pathogens. Further, three plant pathogenic fungi, namely C. capsici, F. oxysporum and P. capsici, were subjected to an in vivo antifungal screening. bDHA at higher concentrations revealed a promising antifungal effect in vivo as compared to the positive control oligochitosan. Furthermore, elaborative study of GC-MS analysis was conducted on bioconverted oil extract of DHA to identify the transformation products present in bDHA. The results of this study indicate that the oil extract of bDHA has potential value of industrial significance to control plant pathogenic fungi.     

Production of lytic enzymes by<Emphasis Type="Italic">Trichoderma</Emphasis> spp. and their effect on the growth of phytopathogenic fungi

The production of β-1,3-glucanases and chitinases by three strains ofTrichoderma in submerged cultures was determined. The synthesis of enzymes was induced by cell wall biopolymers of phytopathogenic fungi (Botrytis cinerea, Fusarium culmorum andF. oxysporum). T. hamatum produced the highest β-1,3-glucanase activity; the most effective inducer of enzyme synthesis was the biomass ofF. oxysporum. All examined strains ofTrichoderma inhibited phytopathogen growth in biotic tests. The diffusion tests showed that the lytic enzymes take part in growth inhibition of phytopathogenic fungi.     

Biodegradation of soil-applied pesticides by selected strains of plant growth-promoting rhizobacteria (PGPR) and their effects on bacterial growth

A laboratory study was conducted to investigate the influence of four PGPR strains on the degradation of five soil applied pesticides and their effects on bacterial growth. Interactions of Bacillus subtilis GB03, Bacillus subtilis FZB24, Bacillus amyloliquefaciens IN937a and Bacillus pumilus SE34 with two concentrations of acibenzolar-S-methyl, metribuzin, napropamide, propamocarb hydrochloride and thiamethoxam in liquid culture and soil microcosm were studied. The degradation of acibenzolar-S-methyl by all PGPR tested in low and high concentration, was 5.4 and 5.7 times, respectively, faster than that in non-inoculated liquid culture medium. At the end of the 72-h liquid cultured experiments, 8–18, 9–11, 15–36 and 11–22% of metribuzin, napropamide, propamocarb hydrochloride and thiamethoxam, respectively, had disappeared from PGPR inoculated medium. Under the soil microcosm experimental conditions, the half-lives of acibenzolar-S-methyl incubated in the presence of PGPR strains spiked at 1.0 and 10.0 mg kg⁻¹ were 10.3–16.4 and 9.2–15.9 days, respectively, markedly lower compared with >34.2 days in the control. From the rest pesticides studied degradation of propamocarb hydrochloride and thiamethoxam was enhanced in the presence of B. amyloliquefaciens IN937a and B. pumilus SE34. Acibenzolar-S-methyl, propamocarb hydrochloride and thiamethoxam significantly increased the PGPR growth. However, the stimulatory effect was related to the level of pesticide spiked.     

Isolation,evaluation and characterization of <Emphasis Type="Italic">Bacillus subtilis</Emphasis> from cotton rhizospheric soil with biocontrol activity against <Emphasis Type="Italic">Fusarium oxysporum</Emphasis>

Present investigation is based on the isolation of Bacillus subtilis from cotton rhizosphere and their evaluation as biocontrol agent against Fusarium oxysporum. The production of extracellular hydrolytic enzyme was studied for determining the antagonism. 43% of 21 isolates were identified under the B. subtilis group on the basis of biochemical characterization. 38% isolates showed competitive activity against Fusarium oxysporum and exhibit more than 50% mycelial inhibition in dual culture bioassay. The pot assay of cotton by seed treatment and soil amendment technique under green house condition showed the competent activity of the isolates in preventing the wilting of cotton seedlings due to F. oxysporum infection. SVI values of 30 day old seedlings indicated that the soil inoculation with B. subtilis BP-2 and seed treatment with B. subtilis BP-9 significantly promoted the growth of cotton seedlings. RAPD profiling revealed the diversity in the Bacillus subtilis group, ranging from 10 to 32%. The discriminative pattern among the isolates belonging to the same species was validated by 16S rDNA partial sequencing which identified them into four different strains of B. subtilis.     

Evaluation of isolates of Trichoderma,Pseudomonas and Bacillus species as treatment for the control of post-harvest fungal rot disease of yam (Dioscorea spp.)

The efficacy of four biological control agents (BCAs): Trichoderma asperellum strain NGT158, T. longibrachiatum strain NGT167, Bacillus subtilis and Pseudomonas fluorescens for the management of post-harvest tuber rot among four yam species, Dioscorea rotundata, D. cayenensis, D. alata, and D. dumetorum was evaluated. Rotted yam tubers were collected across three agroecological zones in Nigeria to isolate six infecting fungal pathogens: Aspergillus niger, Lasiodiplodia theobromae, Rhizoctonia solani, Penicillium oxalicum, Fusarium oxysporum and Sclerotium rolfsii. The BCAs were isolated by serial dilution and rot inhibition of treated tubers was evaluated using destructive sampling method in vivo after six months of storage. Bacillus subtilis was generally most effective, especially when applied 24 h before the inoculation of test pathogens across the four yam species, with percent inhibition that ranged between 47.8 and 81.2%. However, the four BCAs showed good potential in the control of the fungal pathogens causing post-harvest yam rot.     

Bioaccumulation and Biovolatilisation of Pentavalent Arsenic by Penicillin janthinellum, Fusarium oxysporum and Trichoderma asperellum Under Laboratory Conditions

Some fungi are able to control and remediate arsenic (As)-contaminated soil, sediment, or water. Here, we investigate potential accumulation and volatilisation of As by three fungi strains. Results indicated that the highest level of As was accumulated by Penicillin janthinellum with 39.54 μg after 10 days in the culture system amended with 2,500 μg As(V), which represents 50 mg/l As. Fusarium oxysporum showed the highest amount of volatilised As with 304.06 μg after 15 days. The As content in the treated system (filter paper + As + fungi) was significantly higher than that in the control (filter paper + As; filter paper + fungi; filter paper). Trichoderma asperellum and F. oxysporum showed superior abilities for the absorption of extracellular As and accumulation of intracellular As, which accounted for 82.2 and 63.4% of the total accumulated As, respectively. However, P. janthinellum presented an equal distribution of intracellular and extracellular As. Scanning electron microscope (SEM) analysis suggested that little impact on mycelium growth of the three fungal strains was seen after exposure to 50 mg/l As(V) for 5 days, while the growth of fungi in the control was inhibited. The present results demonstrate that P. janthinellum, F. oxysporum, and T. asperellum would be expected to tackle As-contaminated environments.     

Screening of marine actinobacteria for amylase enzymes inhibitors

Amylase inhibitor producing actinobacteria were isolated and characterized from terrestrial environment and there is no much report found from marine environment, hence in the present study, 17 strains isolated from the rhizosphere sediments of mangroves were tested for their amylase inhibition ability. Seawater requirement test for the growth of actinobacteria found that the strains SSR-3, SSR-12 and SSR-16 requires at least 50% and SSR-6 requires at least 25% seawater for their growth. The inhibition activity of both prokaryotic and eukaryotic amylase was tested by using Bacillus subtilis and Aspergillus niger. The maximum amylase activity (40mm) produced by the A. niger was taken as positive control, when the test actinobacteria strains grown in the medium they inhibited amylase activity and was evidenced by the reduction in inhibition zone (14–37 mm) similarly the amylase produced by the Bacillus subtilis was also recorded maximum (35 mm) amylase activity and was taken as positive control, and the test atinobacterial strains reduced enzyme action(12–33 mm) it varied levals. This indicates that the actinobacteria strains were controlled amylase enzyme activity in both the cases. The strain SSR-10 was highly effective and SSR-8 was less effective in inhibiting eukaryotic amylase produced by A. niger. The strain SSR-2 was effective and SSR-6 showed very less effect in inhibiting the prokaryotic amylase produced by the B subtilis.     

Mycoflora of tuber surface of white yam (Dioscorea rotundata poir) and postharvest control of pathogens with Bacillus subtilis

Bacillus subtilis (Enrenberg) Cohn was investigated for its antagonistic properties against surface mycoflora of yam (Dioscorea rotundata Poir) tubers in storage. Yam tubers inoculated with a spore suspension of B. subtilis in potato dextrose broth using a knapsack sprayer showed a drastic reduction in the range and number of mycoflora, including pathogens of the tuber surface in contrast to the control tubers, during the five-month storage period in a traditional yam barn. However, B. subtilis maintained a high frequency of occurrence during the same period. Botryodiploidia theobromae Pat, Fusarium moniliforme Wollen and Reink., Penicillium sclerotigenum Yamamoto, and Rhizoctonia sp. were displaced completely on the treated tubers. The antagonism of B. subtilis was so effective that the normal tuber surface mycoflora was greatly reduced throughout the storage period of five months by a simple initial application of the antagonist.This revised version was published online in October 2005 with corrections to the Cover Date.     

Differentiated pellicle organization and lipopeptide production in standing culture of <Emphasis Type="Italic">Bacillus subtilis</Emphasis> strains

Pellicle formation and lipopeptide production was analysed in standing cultures of different Bacillus subtilis strains producing two or three families of lipopeptides. Despite its ability to produce surfactin, B. Subtilis ATCC 6633 was unable to form stable pellicle at air–water interface. For the ATTC 21332 and ATCC 9943 strains, it was shown for the first time that the lipopeptides were also produced in standing cultures at productivities similar or lower than those obtained when the culture medium is agitated. A differentiated behaviour was observed between these strains in repetitive batch cultures. B. subtilis 9943 formed a wrinkled, thinner and more resistant pellicle than B. subtilis 21332. The structure of the pellicle determined by electron microscopy observations showed that cells of B. subtilis 9943 formed microcolonies whereas those of B. subtilis 21332 rapidly died. Under these conditions, surfactin production by strain 21332 decreased after 2 days whereas it remained stable for B. subtilis 9943 during the 6 days of the cultures. These data indicate that cells of B. subtilis strains growing in pellicle can produce lipopeptides differently depending on their cellular organisation. M. Chollet-Imbert and F. Gancel have contributed equally to the scientific work.     

Suppression of red rot caused by <Emphasis Type="Italic">Colletotrichum falcatum</Emphasis> on sugarcane plants using plant growth-promoting rhizobacteria

Bacterial strains with ability to suppress Colletotrichum falcatum were isolated from the rhizosphere of sugarcane. Thirty nine candidates, chosen on the basis of in vitro antagonism, inhibited C. falcatum growth by 15–65% on test plates. Twenty two isolates causing 50% or more in vitro inhibition were screened for their root colonization ability and biocontrol activity on micropropagated sugarcane plants under greenhouse conditions. Twelve strains suppressed red rot infection in plantlets, but no significant correlation was observed between in vitro pathogen inhibition and in vivo disease suppression. However, isolates showing root colonization over 5.2 log₁₀ CFU g⁻¹ of soil showed highest suppression of C. falcatum and reduction of red rot disease. Six strains with the capability to maintain a significant population in the sugarcane rhizosphere and with a high potential to control red rot were identified by 16S rDNA as Ochrobacterum intermedium NH-5, Pseudomonas putida NH-50, Bacillus subtilis NH-100, Bacillus subtilis NH-160, Bacillus sp NH-217 and Stenotrophomonas maltophilia NH-300.     

Suppression of common root pathogens by helper bacteria and ectomycorrhizal fungi in vitro

 Root pathogens cause considerable loss of tree seedlings in nurseries and are generally difficult to control using conventional methods. Inoculation with ectomycorrhizal fungi may provide some suppression of pathogens. Bacteria (so-called mycorrhization helper bacteria) have been isolated that stimulate mycorrhiza formation on seedling roots and enhance seedling growth; however, their role in pathogen inhibition has not been explored. Four strains of helper bacteria were inoculated together with the ectomycorrhizal fungal species Laccaria bicolor, L. proxima and Suillus granulatus on culture plates to determine inhibition of the pathogens Fusarium oxysporum and Cylindrocarpon sp. Buffered medium was used to rule out acidification of the medium as a mechanism of inhibition. None of the ectomycorrhizal fungal species alone inhibited the growth of Fusarium but all showed slight inhibition of Cylindrocarpon growth. Helper bacterium strain MB3 (Bacillus subtilis) was effective in inhibiting both pathogens and, when inoculated with either L. proxima or S. granulatus, inhibition of Fusarium growth was enhanced over MB3 alone. With Cylindrocarpon, however, only S. granulatus inoculated along with MB3 showed enhanced inhibition over MB3 alone. The other three bacterial strains had little effect on the growth of Fusarium or Cylindrocarpon. More research is necessary to determine if these inhibitory effects are reproducible in situ. Accepted: 23 October 1996     

Screenhouse and Field Persistence of Nonpathogenic Endophytic Fusarium oxysporum in Musa Tissue Culture Plants

Two major biotic constraints to highland cooking banana (Musa spp., genome group AAA-EA) production in Uganda are the banana weevil Cosmopolites sordidus and the burrowing nematode Radopholus similis. Endophytic Fusarium oxysporum strains inoculated into tissue culture banana plantlets have shown control of the banana weevil and the nematode. We conducted screenhouse and field experiments to investigate persistence in the roots and rhizome of two endophytic Fusarium oxysporum strains, V2w2 and III4w1, inoculated into tissue-culture banana plantlets of highland cooking banana cultivars Kibuzi and Nabusa. Re-isolation of F. oxysporum showed that endophyte colonization decreased faster from the rhizomes than from the roots of inoculated plants, both in the screenhouse and in the field. Whereas rhizome colonization by F. oxysporum decreased in the screenhouse (4–16 weeks after inoculation), root colonization did not. However, in the field (17–33 weeks after inoculation), a decrease was observed in both rhizome and root colonization. The results show a better persistence in the roots than rhizomes of endophytic F. oxysporum strains V2w2 and III4w1.     

The synthetic strigolactone GR24 influences the growth pattern of phytopathogenic fungi

Strigolactones that are released by plant roots to the rhizosphere are involved in both plant symbiosis with arbuscular mycorrhizal fungi and in plant infection by root parasitic plants. In this paper, we describe the response of various phytopathogenic fungi to the synthetic strigolactone GR24. When GR24 was embedded in the growth medium, it inhibited the growth of the root pathogens Fusarium oxysporum f. sp. melonis, Fusarium solani f. sp. mango, Sclerotinia sclerotiorum and Macrophomina phaseolina, and of the foliar pathogens Alternaria alternata, Colletotrichum acutatum and Botrytis cinerea. In the presence of this synthetic strigolactone, intense branching activity was exhibited by S. sclerotiorum, C. acutatum and F. oxysporum f. sp. melonis. Slightly increased hyphal branching was observed for A. alternata, F. solani f. sp. mango and B. cinerea, whereas suppression of hyphal branching by GR24 was observed in M. phaseolina. These results suggest that strigolactones not only affect mycorrhizal fungi and parasitic plants, but they also have a more general effect on phytopathogenic fungi.     

Endophytic microorganisms as potential growth promoters of banana

The potential of endophytic microorganisms in promoting the growth of their host plant was determined by artificially introducing five isolates (bacterial and fungal strains: UPM31F4, UPM31P1, UPM14B1, UPM13B8, UPM39B3) isolated from the roots of wild bananas into both healthy and diseased banana plantlets (Berangan cv. Intan). The response of the host plants to endophytic infection was assessed by measuring the change in four growth parameters: plant height, pseudostem diameter, root mass and total number of leaves. The endophytes tested as growth promoters were found to have a significant effect in both healthy and Fusarium-infected (diseased) plantlets. In both experimental systems, the bacterial isolate UPM39B3 (Serratia) and fungal isolate UPM31P1 (Fusarium oxysporum) showed promising growth-promoting properties. Isolate UPM39B3 (Serratia) induced the largest increases in all four growth parameters in healthy plantlets – 3.14 cm (height), 1.12 cm (pseudostem diameter), 2.12 g (root mass) and 1.12 (total number of leaves plant⁻¹) – followed by isolate UPM31P1 (Fusarium oxysporum). The beneficial effect of UPM39B3 (Serratia) and UPM31P1 (Fusarium oxysporum) was also reflected in the diseased plantlets, where pre-treatments with the isolates either singly (T6: UPM31P1; T8: UPM39B3) or in a mixture (T7: UPM31P1 + UPM39B3; T9: UPM14B1 + UPM13B8 + UPM39B3) were able to sustain the growth of plantlets, with significantly higher growth values than those in diseased plantlets that were not infected with endophytes (T10: FocR4). These results demonstrate the economic significance of these endophytic isolates, particularly UPM39B3 (Serratia) and UPM31P1 (Fusarium oxysporum), both as potential growth promoters of banana and as agents rendering tolerance towards Fusarium wilt as a strategy in the management of Fusarium wilt of banana via improved vegetative growth.     

Antagonistic actions of Pythium oligandrum and Trichoderma harzianum against phytopathogenic fungi (Fusarium oxysporum and Pythium ultimum var. ultimum)

Abstract

The possible biological control of damping-off fungi, Fusarium oxysporum and Pythium ultimum by Pythium oligandrum or Trichoderma harzianum was in vitro investigated. Results of comparing the antagonistic activity of P. oligandrum and T. harzianum in dual plates against the tested phytopathogens indicated different degrees of antagonism. After 12 days of incubation colony of the phytopathogenic fungus was completely overgrown by the antagonist, except for the interaction between T. harzianum and F. oxysporum which showed no overgrowth or any hyphal penetration by the antagonist. However, growth and proliferation of F. oxysporum colony was repressed. T. harzianum and P. oligandrum produced chitinase and β-1,3-glucanase when they were grown on liquid culture medium supplemented with chitin or fungal dried mycelium as a sole carbon source, and enzyme production was higher by T. harzianum comparing with P. oligandrum under the same condition. Fungal dried mycelium of F. oxysporum was the most selective carbon source for enzyme production, on the other hand, chitinase production was significant locked when P. ultimum dried mycelium was used as a carbon source. Production of volatile compounds by P. oligandrum or T. harzianum against F. oxysporum and P. ultimum was examined using the inverted plates method. F. oxysporum was inhibited by the antagonist volatile compounds and it is inhibited 100% by increasing the amount of inoculum size. Production of potential biocontrol agents provided with economically features and working under field conditions are recommended.     

Glucosinolate degradation by Aspergillus clavatus and Fusarium oxysporum in liquid and solid-state fermentation

Two fungal strains, Aspergillus clavatus II-9 and Fusarium oxysporum @ 149, proved to be capable of degrading sinigrin and sinalbin. During the degradation of sinigrin by whole cells of the Aspergillus strain, allylcyanide accumulated in the liquid incubation mixture. After a maximum concentration had been reached, the concentration of allylcyanide decreased as a result of its instability in the medium used. Incubation of cell-free extracts with sinigrin resulted in accumulation of glucose and allylisothiocyanate, suggesting that myrosinase is involved. Experiments with intact cells and cell-free extracts indicate the formation of an as yet unknown intermediate. When sinigrin was degraded by the Aspergillus strain in mustard seed meal under solid-state fermentation (SSF) conditions, no accumulation of allylcyanide or allylisothiocyanate was measured. Degradation of sinigrin by F. oxysporum @ 149 did not result in accumulation of intermediates, neither in liquid incubation mixtures nor in mustard seed meal under SSF conditions. Sinigrin was not degraded during incubation with cell-free extracts of F. oxysporum @ 149. Degradation of sinalbin by A. clavatus and F. oxysporum was measured during fermentation of yellow mustard seed meal under SSF conditions. Both fungi are useful for laboratory-scale SSF of mustard seed meal, thus opening new perspectives for a cost effective detoxification process for raw feed materials. Correspondence to: J. P. Smits     

Decomposition of cellulose strips in relation to climate,litterfall nitrogen,phosphorus and C/N ratio in natural boreal forests

Isolates of Alternaria alternata, Botrytis cinerea, Fusarium oxysporum, Penicillium sp., Rhizoctonia solani, Stemphylium sp., Thielaviopsis basicola, and Verticillium dahliae were cultured on potato–dextrose agar (PDA), barley-sand and alfalfa-sand substrates in petri-dish or in column microcosms. N-mineralization by fungi and fungal-feeding nematodes in combination or fungi alone was assessed. Numbers of Aphelenchus avenae or Aphelenchoides composticola supported by the fungi were measured every 7 days. Times for full colonization of the substrates by fungi ranged from 5 to 15 days. Rhizoctonia solani and B. cinerea on PDA supported the largest A. avenae and A. composticola populations, respectively. Penicillium sp. was a nonhost for A. composticola and A. avenae. Rhizoctonia solani, B. cinerea, V. dahliae, and F. oxysporum supported significantly more nematodes than the other four fungal species. The ranked order of fungi based on the amount of N mineralized in columns free of nematodes was A. alternata (with a rate of 0.052 μg N/g-sand per day), Stemphylium sp., V. dahliae, T. basicola, B. cinerea, F. oxysporum, R. solani, and Penicillium sp. (with a rate of 0.0045 μg N/g-sand perday). The presence of A. avenae resulted in significant increases in mineral N, compared to nematode-free columns colonized by F. oxysporum, R. solani, and T. basicola alone. The presence of A. composticola resulted in significant increases in mineral N, compared to nematode-free columns colonized by A. alternata, B. cinerea, F. oxysporum, and R. solani alone. There was more mineral N incolumns in the presence of A. composticola than A. avenae in most cases. This revised version was published online in August 2006 with corrections to the Cover Date.