Fusarium species in soil,thatch and crowns of Poa pratensis turfgrass treated with fungicides

Little information exists concerning the effects of long-term fungicide programs on the qualitative and quantitative characteristics of fusaria in turfgrasses and in turfgrass soils. These effects were studied on Poa pratensis L, turfgrasses at three locations, using 14 fungicides, one nematicide, and five mixed fungicide programs. Some fungicides increased the numbers of fusaria in soil and thatch, some had no effect, and others greatly reduced the numbers. Changes in Fusarium species' compositions occurred independently from the changes in propagule numbers. The proportion of Fusarium-colonized turfgrass crowns was generally higher in fungicide-treated than in nontreated turfgrasses.     

Earthworms promote the reduction of Fusarium biomass and deoxynivalenol content in wheat straw under field conditions

A field experiment was conducted to elucidate ecosystem services provided by earthworms on the repression of phytopathogenic and toxinogenic fungi. The study focussed on decomposing Fusarium culmorum-infected and deoxynivalenol (DON)-contaminated wheat straw remaining on the soil surface as part in conservation tillage. Mesocosms were established in the topsoil of a winter wheat field located in Northern Germany, where conservation tillage has been practised for 20 years. Besides a non-earthworm treatment, two earthworm species were inoculated in the mesocosms either separately or combined: Lumbricus terrestris (anecic, detritivorous) and Aporrectodea caliginosa (endogeic, geophagous). The earthworms were exposed either to artificially Fusarium-infected wheat straw highly contaminated with DON or to non-infected straw serving as a control. The experiment was conducted during an eight week period after harvest from mid August to mid October. For both species, the artificially Fusarium-infected and DON-contaminated wheat straw was a more attractive food source than the non-infected control. In contrast to A. caliginosa, L. terrestris incorporated infected straw faster into the soil compared to control straw. Furthermore, the reduction of Fusarium biomass and DON concentration in wheat straw was significantly higher in the presence of L. terrestris than in treatments with A. caliginosa and without earthworms. Here, no significant differences could be measured between the Fusarium biomass and DON concentration in wheat straw. A. caliginosa seems not to be relevant for the reduction of Fusarium biomass and DON concentration. We concluded that amongst earthworms, anecic detritivorous species are the drivers to compensate possible negative consequences (like crop infection) of conservation tillage. They take an important role in the control of phytopathogenic and toxinogenic fungi surviving on plant residues and in the degradation of their mycotoxins.     

Non-pathogenic Fusarium strains protect the seedlings of Lepidium sativum from Pythium ultimum

Two root-colonizing Fusarium strains, Ls-F-in-4-1 and Rs-F-in-11, isolated from roots of Brassicaceae plants, induced the resistance in Lepidium sativum seedlings against Pythium ultimum. These strains caused an increase in the content of benzyl isothiocyanate, and of its precursor glucotropaeolin, in the roots of the host plants. The increased isothiocyanate content is one of the factors contributing to the resistance of L. sativum against P. ultimum. To be transformed into the fungitoxic compound benzyl isothiocyanate, glucotropaeolin has to be hydrolyzed by myrosinase, which can be produced either by plants or microorganisms. The Fusarium strain Ls-F-in-4-1 has a myrosinase activity but the strain Rs-F-in-11 has not. These results suggest that both strains are able to trigger the metabolic pathway leading to benzyl isothiocyanate production in the plant. In the case of the myrosinase-negative strain Rs-F-in-11, hydrolyzation into isothiocyanate is only due to the myrosinase activity of the plant, and in the other case, the myrosinase produced by the strain Ls-F-in-11 also would contribute to the production of isothiocyanate. This paper reports a new mode of action of non-pathogenic Fusarium strains in controlling P. ultimum.     

Application of bio-organic fertilizer can control Fusarium wilt of cucumber plants by regulating microbial community of rhizosphere soil

Fusarium wilt, caused by Fusarium oxysporum f. sp. cucumerinum J. H. Owen, results in considerable yield losses for cucumber plants. A bio-organic fertilizer (BIO), which was a combination of manure composts with antagonistic microorganisms, and an organic fertilizer (OF) were evaluated for their efficiencies in controlling Fusarium wilt. Application of the BIO suppressed the disease incidence by 83% and reduced yield losses threefold compared with the application of OF. Analysis of microbial communities in rhizosphere soils by high-throughput pyrosequencing showed that more complex community structures were present in BIO than in OF treated soils. The dominant taxonomic phyla found in both samples were Proteobacteria, Firmicutes, Actinobacteria and Acidobacteria among bacteria and Ascomycota among fungi. Abundance of beneficial bacteria or fungi, such as Trichoderma, Hypoxylon, Tritirachium, Paenibacillus, Bacillus, Haliangium and Streptomyces, increased compared to the OF treatment, whereas the soil-borne pathogen, Fusarium, was markedly decreased. Overall, the results of this study demonstrate that the application of the BIO was a useful and effective approach to suppress Fusarium wilt and that the high-throughput 454 pyrosequencing was a suitable method for the characterization of microbial communities of rhizosphere soil of cucumber.     

Electron microscopical studies of chlamydospores of Fusarium solani F. cucurbitae formed in natural soil

Macroconidia of Fusarium solani f. cucurbitae were placed into natural soil, incubated for various times, recovered and examined by light and transmission electron microscopy. Lysis of macroconidia and formation of chlamydospores were studied and the fine structure of these propagules and their associated microflora was investigated. The two most obvious features of chlamydospore morphology were the sloughing of outer wall layers and the accretion of micro-fibrillar elements adjacent to the plasmalemma. Chlamydospore formation was partially suppressed by addition of nitrogenous compounds to the soil.     

Mycoparasitism by Pythium oligandrum and P. acanthicum

Isolates of the reported mycoparasites Pythium oligandrum, P. acanthicum and P. periplocum markedly reduced growth and cellulolysis by Botryotrichum piluliferum, grew rapidly across agar plates precolonized by Phialophora radicicola var radicicola (sensu Deacon) and, where tested (not P. periplocum), were non-pathogenic towards higher plants. Isolates of P. echinulatum, P. mamillatum. P. megalacanthum, P. spinosum, P. ultimum and one isolate of P. acanthicum behaved differently from the mycoparasites and could, themselves, be placed in two groupings in these tests. It is suggested that the ability or otherwise to grow on Phialophora-precolonized agar plates may help to distinguish broad biological groupings within the genus Pythium, but these groupings may cut across conventional taxonomic ones.One isolate of P. acanthicum was tested for its effects on a range of cellulolytic fungi: it reduced their growth to different extents, as did P. oligandrum.Plates of potato-dextrose agar precolonized by Phialophora radicicola were used to isolate selectively P. oligandrum and similar fungi from soils, but the use of hemp seed baits in conjunction with precolonized plates was less selective for these fungi.Straw pieces precolonized by P. oligandrum and buried in soil decomposed at the same rate as virgin straws or those precolonized by P. ultimum or Mucor hiemalis. Subsequently, Stachyholrys atra appeared to colonize straws more frequently from soil, and Fusarium spp. less so, in the presence of P. oligandrum than in its absence. In the laboratory, P. oligandrum was antagonized by Slachyholrys, whereas Fusarium spp. were frequently overgrown by the Pythium.     

Germination and hyphal growth of Glomus caledonicum on water agar containing benomyl

Concentrations of benomyl greater than 10 ng ml^?1 inhibited germination and hyphal growth of the vesicular-arbuscular mycorrhizal fungus Glomus caledonicum on water agar. This effect was attributable to only the activity of methyl-2-benzimidazole carbamate, one of the two hydrolysis products of benomyl. Hyphal growth but not germination was affected by the pH of water agar buffered with 10 mm 2-(N-morpholino)ethane sulphonic acid.     

Effets de quelques pesticides sur la colonisation d'un substrat par le Trichoderma harzianum rifai en presence des autres champignons du sol

The effects of some pesticides upon substrate colonization by Trichoderma harzianum Rifai in the presence of other soil fungi.A herbicide (propyzamide) and 5 fungicides (benomyl, quintozene, vinclozolin. thiram, prothiocarb) were added at various concentrations to two Trichoderma harziunum-enriched garden soils. After thorough mixing, followed by solidifying with water agar, discs of these mixtures were cut with a cork-borer and transferred to a non specific nutrient medium (PDA). The development of T. harzianum on the agar, around each implant, was estimated after incubation for 1 week at 28°C.In the presence of other soil fungi, T. harzianum development was scarcely or not at all modified by prothiocarb, propyzamide and quintozene. It was highly depressed by benomyl. Vinclozolin was only slightly inhibitory. In our experimental conditions, T. harzianum development was strongly enhanced by thiram, even at a high dosage (300 μg ml^?1).The limits and the advantages of the method are estimated and the results are discussed in view of practical applications.     

Effects of Arbuscular Mycorrhizal Colonization on Microbial Community in Rhizosphere Soil and Fusarium Wilt Disease in Tomato

Fusarium wilt is caused by soil-borne pathogen Fusarium oxysporum. Tomato (Lycopersicon esculentum Mill.) is susceptible to Fusarium oxysporum f. sp. lycopersici race 1 and was infected with wilt disease. A pot experiment was conducted to investigate effects of inoculating arbuscular mycorrhizal (AM) fungus (Glomus etunicatium) on the microbial community in the rhizosphere soil and Fusarium wilt in tomato (cv. Oogatafukuju). The results indicated that AM fungal inoculation suppressed the Fusarium number in the rhizosphere soil of tomato and decreased the Fusarium wilt disease index. Compared to the control, AM fungal inoculation increased the actinomycete number but increased bacterial number. Bacterial and fungal numbers were high but actinomycetes number was low when tomato basal stems became discolored brown. Fusarium inoculation significantly suppressed development of AM colonization and decreased polyphenol oxidase (PPO) activity in leaves and roots of tomato. Inoculation with AM fungi and Fusarium maintained high PPO activity in leaves and roots. The AM colonization increased root growth of tomato, whereas Fusarium inoculation had no significant effect on tomato growth. These findings suggest that because AM fungal inoculation changes microbial communities and enhances PPO activity, it should suppress occurrence of Fusarium wilt in tomato.     

Relationship of root necrosis potential of soil to asparagus (Asparagus officinalis) decline in Switzerland

Summary The possible relationship between asparagus decline and the root necrosis potential (RNP) of soil was investigated for 11 asparagus and 4 nonasparagus soils. Asparagus seedlings were grown in each soil in climate chambers. A root necrosis index was used to determine RNP. RNP was correlated with the decline observed in the field for seven white asparagus crops, but not for three white and one green asparagus crops. Low RNP values were obtained from the soils not planted to asparagus.Fusarium oxysporum andF. oxysporum var. redolens accounted for 7070 of the fungi isolated from necrotic roots in RNP tests. BothFusarium were obtained from all soils. AllF. oxysporum andF. oxysporum var. redolens isolates tested were highly virulent on asparagus seedlings in inoculation experiments.F. solani, Penicillium verrucosum var.corymbiferum, andRhizoctonia violacea were isolated less frequently but were also highly virulent. It is concluded that soil RNP indicated a risk of decline caused by fusaria, but other soil factors were likely to be involved in the aetiology of decline. The study of these factors is necessary to develop a method for the prognosis of decline associated with fusaria.     

盐胁迫下两种草坪草膜脂过氧化和抗氧化酶比较

Salinity stress is a major factor limiting the growth of turfgrass irrigated with recycled wastewater. The change in lipid peroxidation in terms of malondialdehyde (MDA) content and the activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxide (APX) and glutathione reductase (GR) in the shoots and roots of Kentucky bluegrass and tall fescue were investigated under salinity stress. Plants were subjected to 0, 50, 100, 150 and 200 mmol L 1 NaCl for 40 d. The MDA content under salinity stress was lower in tall fescue than in Kentucky bluegrass in both shoots and roots. Activities of SOD in the shoots of both species increased with salinity stress. The activities of CAT and APX decreased in Kentucky bluegrass, but no significant difference in the activities of CAT and APX was observed in tall fescue. The activities of SOD, CAT and APX in the shoots of tall fescue were higher than those in Kentucky bluegrass. In the roots of Kentucky bluegrass, SOD and GR activities increased and CAT and APX activities decreased in comparison with the control. In the roots of tall fescue, salinity increased the activities of SOD, CAT, and APX. These results suggested that tall fescue exhibited a more effective protection mechanism and mitigated oxidative stress and lipid peroxidation by maintaining higher SOD, CAT and APX activities than Kentucky bluegrass.     

Soil temperature,mycorrhizal infection and nodulation of Medicago truncatula and Trifolium subterraneum

Establishment of vesicular-arbuscular mycorrhizal fungi in plant roots involves a pre-infection phase of propagule germination, hyphal growth and appressorium formation, followed by growth of the fungus within the root. The effect of soil temperature on the pre-infection stage was examined by counting the numbers of fungal “entry-points” on the main roots of Medicago truncatula and Trifolium subterraneum, grown at soil temperatures of 12°, 16°, 20° and 25°C for periods up to 12 days. Increased root temperature was positively associated with increased numbers of “entry-points”. This effect was more marked between 12° and 16°C than at higher temperatures, as shown by comparing plants at the same stage of development (emergence of spade leaf) and by calculating the results as entry points per cm root.The first root nodules appeared sooner at higher temperatures (20° and 25°), but subsequent development of nodules (measured as nodule number and aggregate volume of nodules per plant, up to 21 days) was best at 16°C for both host Rhizobium combinations in non-sterile and autoclaved soil. There was no evidence that competition between mycorrhizal fungi and Rhizobium for infection sites occurred.A method of obtaining numbers of infective propagules of vesicular-arbuscular mycorrhizal fungi in soil is described.     

Diurnal patterns of nitrate assimilation in Kentucky bluegrass

Kentucky bluegrass (Poapratensis L.) is a major C₃‐type forage and turfgrass, but it is less efficient than many grasses in utilizing nitrogen(N). To determine how this grass can accommodate its greater N need, diurnal patterns of nitrate reductase activity (NRA) and nitrite reductase activity (NiRA) in its leaves and roots were examined and compared with those in barley (Hordeum vulgare L.). Plants were grown under greenhouse or growth room conditions and assayed for NRA and NiRA by optimized in vivo methods. The diurnal patterns of NRA and NiRA indicated that Kentucky bluegrass could assimilate nitrate during the night at rates greater than or similar to those during the day. Leaf NRA of Kentucky bluegrass was minimal approximately 4 and 10 h after illumination commenced and increased at night. The diurnal pattern of leaf NRA among Kentucky bluegrass cultivars did not differ significantly. In roots, NRA of Kentucky bluegrass was high in the morning and decreased sharply during the afternoon and evening, but increased again late at night. Unlike Kentucky bluegrass, barley exhibited greater leaf NRA during the day than during the night and exhibited the greatest activity 6 or 10 h after illumination commenced. In both species, the equilibrium leaf nitrate pool was 20 to 30 times larger than the ammonium pool and 3, 000 to 13, 000 times larger than the nitrite pool. Leaf nitrate pool size showed a diurnal pattern complementary to that of leaf NRA. Our results suggest that a nighttime N use strategy might exist in Kentucky bluegrass.     

Host status of 10 fungal isolates for two nematode species, Filenchus misellus and Aphelenchus avenae

The classification of nematodes in the family Tylenchidae into plant parasites, plant associates or fungal-feeders for community analyses, have been much discussed by nematode ecologists. For an appropriate classification, fungal-feeding habits in the family need to be studied. To evaluate the host status of 10 fungal isolates for Filenchus misellus (Tylenchidae) and Aphelenchus avenae (Aphelenchida, Aphelenchidae), population growth rates, body length and width and sex ratios of the nematodes were measured after 40-day culture on fungal colonies at 25 °C. For F. misellus, the fungi determined as good hosts were two Basidiomycota fungi (Agaricus bisporus, Coprinus cinereus), three Ascomycota fungi (Chaetomium cochlioides, Chaetomium funicola, Chaetomium globosum) and a plant-pathogenic fungus (Rhizoctonia solani) on the basis of nematode population growth rate and female body length. Interestingly Pleurotus ostreatus, known as a predaceous fungus for the other nematodes, was also a good host for F. misellus. While, for A. avenae, good hosts were four plant-pathogenic fungi (Fusarium oxysporum f. sp. conglutinans, F. oxysporum f. sp. cucumerinum, Pythium ultimum, R. solani) and A. bisporus. A. avenae was trapped and preyed upon by Pleurotus hyphae. In F. misellus, males were 7-21% of adults, but the ratio did not correlate significantly with the population growth rate. In A. avenae, no male occurred. Differences in habitat preference between Filenchus and Aphelenchus were explained on the basis of the host status and habitat preferences of the tested fungi.     

Soils suppressive to Gaeumannomyces graminis var. tritici: Induction by other fungi

Rhizosphere soils were obtained from wheat plants growing in fumigated soil inoculated with one of the pathogens, Rhizoctonia solani, Gibberella zeae, Fusarium culmorum, Cochliobolus sativus and Pythium irregulare or one of the non-pathogenic fungi, Gaeumannomyces graminis var. graminis (Ggg) or a Phialophora-like fungus (Plf). Using a pot bioassay, these soils were tested for suppression of Gaeumanomyces graminis var. tritici (Ggt) and the fungus involved in the initial induction. G. zeae was the only fungus that induced suppression to Ggt and to itself. Ggg but not Plf induced suppression to Ggt although both fungi induced suppression to themselves.Fungi capable of inducing suppression of take-all have two characteristics in common, they induce suppression of themselves and their saprophytic survival is restricted to organic matter.     

Pre-symbiotic and symbiotic interactions between Glomus intraradices and two Paenibacillus species isolated from AM propagules. In vitro and in vivo assays with soybean (AG043RG) as plant host

Two indole-producing Paenibacillus species, known to be associated with propagules of arbuscular mycorrhizal (AM) fungi, were examined for their mycorrhization helper bacteria activity at pre-symbiotic and symbiotic stages of the AM association. The effects were tested under in vitro and in vivo conditions using an axenically propagated strain of the AM fungus Glomus intraradices and Glycine max (soybean) as the plant host. The rates of spore germination and re-growth of intraradical mycelium were not affected by inoculation with Paenibacillus strains in spite of the variation of indole production measured in the bacterial supernatants. However, a significant promotion in pre-symbiotic mycelium development occurred after inoculation of both bacteria under in vitro conditions. The Paenibacillus rhizosphaerae strain TGX5E significantly increased the extraradical mycelium network, the rates of sporulation, and root colonization in the in vitro symbiotic association. These results were also observed in the rhizosphere of soybean plants grown under greenhouse conditions, when P. rhizosphaerae was co-inoculated with G. intraradices. However, soybean dry biomass production was not associated with the increased development and infectivity values of G. intraradices. Paenibacillus favisporus strain TG1R2 caused suppression of the parameters evaluated for G. intraradices during in vitro symbiotic stages, but not under in vivo conditions. The extraradical mycelium network produced and the colonization of soybean roots by G. intraradices were promoted compared to the control treatments. In addition, dual inoculation had a promoting effect on soybean biomass production. In summary, species of Paenibacillus associated with AM fungus structures in the soil, may have a promoting effect on short term pre-symbiotic mycelium development, and little impact on AM propagule germination. These findings could explain the associations found between some bacterial strains and AM fungus propagules.     

Evaluating soil sodium indices in soils of volcanic nature conducive or suppressive to Fusarium wilt of banana

The ability of three soil Na indices to predict soil conduciveness or suppressiveness to disease caused by the soil fungus Fusarium oxysporum f. sp. cubense was evaluated in seven banana plantations from the Canary Islands (Spain). These indices were exchangeable sodium percentage (ESP), soluble Na (SS₀) and sodium adsorption ratio (SAR₀) in 1:2.5 soil-water extracts (SAR_w and total cationic concentration (TCC_w)=0. Sodium selectivity coefficients (K_G0,K₀) and TCC₀ were calculated from soil exchange and solution data. The effects of ESP, SAR₀, SS₀, TCC₀, K_G0 and K₀ on soil available iron (Fe extracted from soil by DTPA) and aggregate stability in water (water-stable aggregates (WSA), 200-2000 μm) were also studied. Our results showed that SAR₀ calculated using cationic concentrations in 1:2.5 extracts might be a good indication of a relationship between SS₀ and soluble divalent cations in conducive and suppressive volcanic soils to Fusarium. Both TCC₀ and dispersion-flocculation concentrations seem to be not linked to soil suppressiveness or conduciveness to Fusarium wilt. These results suggested that soil physical properties seem to be not controlled by Na behaviour in these type of soils and, therefore, sodicity and salinity should not be a problem from a physical point of view. Moreover, SS₀ and SAR₀ were always greater in suppressive areas than in conducive areas. SAR₀ was significantly correlated with SS₀ but correlations between ESP against SS₀ and SAR₀ were weak. For SAR₀ values above 2.5 (mmol_c l⁻¹)^1/2 and ESP values below 15%, the exchangeable Na did not seem to be related to the capacity of suppressive areas to release more Na to soil solution. Larger values of SS₀ were observed in suppressive areas for these values of SAR₀ and ESP. It implies a lower quantity of soluble Na salts in conducive samples. A high Na salt content in soil can produce an increase of soil pH, which exerts a negative influence on available Fe release to soil solution. A clear separation between conducive and suppressive samples from relations between SS₀ and SAR₀ against WSA and Fe-DTPA showed that SS₀ and SAR₀ can be satisfactory indices to study the influence of Na concentrations on the incidence of Fusarium wilt. The mass of WSA increase in conducive areas might be favoured by the smaller amounts of soil solution Na found in these samples. In conclusion, our data provide evidence that release of Na to soil solution could favour soil suppressiveness to Fusarium wilt limiting soil aggregation and the availability of Fe, at least in soils of volcanic nature that are not affected by salinity or sodicity processes.     

Cause and duration of mustard incorporation effects on soil-borne plant pathogenic fungi

Two fungal plant pathogens, Rhizoctonia solani AG 2-2 and Fusarium oxysporum f.sp. lini, were studied in relation to general responses of soil fungi and bacteria following incorporation of Brassica juncea. Our aim was to understand to what extent the changes in the biological and physicochemical characteristics of the soil could explain the effects on the studied pathogens and diseases, and to determine the temporal nature of the responses. Short-term effects of mustard incorporation (up to 4 months) were investigated in a microcosm experiment, and compared with a treatment where composted plant material was incorporated. In a field experiment, the responses were followed up to 11 months after removal or incorporation of a mustard crop. In general, responses in the variables measured changed more after incorporation of fresh mustard material than after addition of similar amounts of composted plant material (microcosms) or after removal of the mustard crop (field). The soil inoculum potential of R. solani AG 2-2 decreased directly after incorporation of mustard, but increased later to disease levels above those in the untreated soil. Neither of these effects could be explained by changes in the population density of R. solani AG 2-2. Fusarium spp. were less influenced, although an increase in the suppressiveness to Fusarium wilt was observed after mustard incorporation as compared with the treatment where mustard was removed. The microbial responses to mustard incorporation were more pronounced for bacteria than for fungi. After an initial substantial increase, the bacterial density decreased but remained above the levels in the control treatment throughout the experimental periods. The bacterial community structure was modified up to 8 months after mustard incorporation. We conclude that incorporation of fresh mustard influences soil microbial communities, especially the bacteria, and has a potential to control the pathogenic activity of R. solani 2-2 on a short-term perspective. The time dependency in microbial responses is important and should be taken into consideration for the evaluation of the potential of Brassicas to control plant disease on a field scale.     

Amoebae from a take-all suppressive soil which feed on Gaeumannomyces graminis tritici and other soil fungi

Amoebae were isolated from soil of the Waite Institute permanent pasture plot which is suppressive to take-all of wheat. Nine species of amoebae belonging to eight genera were tested for their mycophagy against Gaeumannomyces graminis var. tritici, Cochliobolus sativus and Phytophthora cinnamomi. Members of the genera, Gephyramoeba, Mayorella, Saccamoeba, Thecamoeba and an unidentified species of the order Leptomyxida, were mycophagous. Feeding of mycophagous amoebae and their ability to perforate and lyse melanized propagules of fungi are discussed.