Roles of low pH, carbon and inorganic nitrogen source use in chlamydospore formation by Fusarium solani.

Citrate and malate were poorer sources of exogenous carbon than several hexose, pentose, or disaccharide sugars for supporting macroconidial germination by Fusarium solani at high conidial density (1 X 10(5) condia/ml). Only citrate, however, failed to block chlamydospore morphogenesis to a degree comparable to glucose or other readily used sugars. Mostly immature chlamydospores were formed in the presence of citrate. At low conidial density (5 X 10(3) conidia/ml), exogenous carbon-independent macroconidial germination and subsequent rapid chalmydospore formation on germ tubes was not inhibited by ammonium or nitrate nitrogen. The citrate-phosphate buffered, low pH (4.0) medium of Cochrane induced more immature chlamydospore formation by F. solani than a pH 6.0 medium, but few mature chlamydospores were formed in either medium. Condensation of hyphal cytoplasm into developing chlamydospores, a character typical of chlamydospore formation, did not occur extensively and macroconidia, hyphae, and immature chlamydospores stained deeply with Sudan III, suggesting lipid biosynthesis. This inhibition of chlamydospore maturation may be due partly to nitrogen deficiency imposed by the high C:N ratio of the medium and to the presence of citrate. Only vesiculate hyphal cells were formed by F. solani f. sp. phaseoli in both media. Field soils to which the clone of F. solani used is indigenous had mean pH values ranging from 5.2 to 6.0.     

Diversity of antifungal actinomycetes in various vegetative soils of Korea

Diversity of actinomycetes and their antifungal activities against some plant pathogenic fungi were examined in various vegetative soils from 14 different sites in the western part of Korea. Actinomycete counts ranged from 1.17 x 10(6) to 4.20 x 10(6) cfu x g(-1) dried soil. A total of 1510 actinomycetes were isolated from the soil samples. Streptomyces was predominant in soils with a pH range of 5.1-6.5, 9.1-13.0% moisture, and 9.1-11.0% organic matter. Most Micromonospora, Dactylosporangium, and Streptosporangium were distributed in soils with pH 4.0-5.0, 2.0-9.0% moisture, and 4.0-7.0% organic matter. Actinomadura and nocardioform actinomycetes were abundant in soils with pH 4.0-5.0 and 13.1-20.0% moisture and with 9.1-11.0 and 4.0-7.0% organic matter, respectively. Populations of Streptomyces were predominant in all the soils, but were highest in grassland and lowest in mountain-forest soils. Micromonospora was most abundant in pepper-field soil and nocardioform actinomycetes were highest in rice paddy field soil. Dactylosporangium was predominant in lake-mud sediments and pepper-field soil, Streptosporangium in lake-mud sediments, and Actinomadura in mountain-forest soil. Antifungal actinomycetes were abundant in orchard soil and lake mud. More than 50% of antifungal isolates from most soils were classified as genus Streptomyces. Actinomycete isolates that showed strong antifungal activity against Alternaria mali, Colletotrichum gloeosporioides, Fusarium oxysporum f.sp. cucumerinum, and Rhizoctonia solani were predominant in pepper-field soils, whereas those against Magnaporthe grisea and Phytophthora capsici were abundant in radish-field soils.     

Soil fungi of some low-altitude desert cotton fields and ability of their extracts to inhibit Aspergillus flavus

Soil is presumed to be a major source of inoculum for Aspergillus flavus which contaminates cottonseed and produces the potent carcinogen, aflatoxin. Little is known about the mycoflora of the low desert soils of cotton fields where aflatoxin is a chronic problem. In this study, soils from cotton fields in southwestern Arizona and southeastern California were assayed for filamentous fungi. Forty-two taxa, predominantly in the genera Aspergillus, Penicillium and Fusarium, were isolated. To determine whether or not compounds produced by these fungi could be potential inhibitors of A. flavus, extracts of strains of each taxon were tested for their ability to inhibit growth of A. flavus. Twelve taxa produced compounds inhibitory to A. flavus, including several strains of Fusarium solani, Penicillium vinaceum and Aspergillus auricomus. This revised version was published online in June 2006 with corrections to the Cover Date.     

Differentiation inside multicelled macroconidia of Fusarium culmorum during early germination

Multicelled conidia are formed by many fungal species, but germination of these spores is scarcely studied. Here, the germination and the effects of antimicrobials on multicompartment macroconidia of Fusarium culmorum were investigated. Germ-tube formation was mostly from apical compartments. The intracellular pH (pH(in)) of the different individual cells of the macroconidia was monitored during germination. The pH(in) varied among different compartments and during different stages of germination. The internal pH was lowest in ungerminated cells and rose during germ-tube formation and was highest in new germ tubes. Antifungal compounds affect the pH(in) and differentiation of the conidia. The pH(in) inside the macroconidial compartments was lowered very fast in the presence of nystatin (1 and 4 microg/ml). At sublethal doses (0.3 microg/ml), the apical compartments were preferentially targeted showing lower pH(in) values. The reduced germination capacity of apical compartments under these conditions was compensated by an increased germination capacity of middle compartments.     

温度对茄病镰刀菌生长情况及产孢量的影响

目的 探讨不同孵育温度对茄病镰刀菌生长状态的影响,为获得大量饱子寻找较好方案.方法 茄病镰刀菌按孵育温度不同分为6组,A组、B组、C组、D组、E组分别于20℃,25℃,30℃,35℃,37℃温箱黑暗孵育7d,F组于35℃温箱黑暗孵育3d再于25℃温箱黑暗孵育至7d,各组在培养24h,3d,5d,7d时分别用游标卡尺测量...     

Isolation and Characterization of Microsporum gypseum Lysosomes: Role of Lysosomes in Macroconidia Germination

Three types of lysosomes containing either acid protease, alkaline protease, or phosphodiesterase were isolated from a Microsporum gypseum macroconidial homogenate on Ficoll gradients. The acid protease was contained in an assimilative lysosome since its activity was affected by the complexity of the exogenous nitrogen source. Ultracentrifugation and electron microscopy revealed that the alkaline protease-containing vesicles were associated with the spore coat material prior to macroconidial germination. During macroconidial germination, zones of spore coat hydrolysis were seen surrounding these vesicles. Other larger vesicles, believed to contain the phosphodiesterase, were also observed in the spore coat during macroconidial germination.     

Non-pathogenic Fusarium solani represses the biosynthesis of nematicidal compounds in vitro and reduces the biocontrol of Meloidogyne javanica by Pseudomonas fluorescens in tomato

AIMS: The aim of the present investigation was to determine the influence of various Fusarium solani strains on the production of nematicidal agent(s) in vitro and biocontrol of Meloidogyne javanica in tomato by Pseudomonas fluorescens strains CHA0 and CHA0/pME3424. METHODS AND RESULTS: Culture filtrates (CF) of P. fluorescens strain CHA0 and its diacetylphloroglucinol-overproducing derivative CHA0/pME3424 caused substantial mortality of M. javanica juveniles in vitro. Bacterial growth medium amended with the growth medium of F. solani repressed the nematicidal activity of the bacteria. Methanol extract of F. solani CF resulting from Czapek's Dox liquid (CDL) medium without zinc amendment repressed the nematicidal activity of the bacteria while the CF obtained from CDL medium amended with zinc did not. Conidial suspension of F. solani strain Fs5 (repressor strain for the biosynthesis of nematicidal compounds in P. fluorescens) reduced biocontrol potential of the bacterial inoculants against M. javanica in tomato while strain Fs3 (non-repressor) did not. CONCLUSIONS: Fusarium solani strains with increased nematicidal activity repress the biosynthesis of nematicidal compounds by P. fluorescens strains in vitro and greatly alter its biocontrol efficacy against root-knot nematode under natural conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: Fusarium solani strains are distributed worldwide and found in almost all the agricultural fields which suggest that some mycotoxin-producing strains will also be found in almost any soil sample taken. Besides the suppressive effect of these metabolite-producing strains on the production of nematicidal compound(s) critical in biocontrol, F. solani strains may also affect the performance of mycotoxin-sensitive biocontrol bacteria effective against plant-parasitic nematodes.     

Detection of mutagens produced by fungi with the Salmonella typhimurium assay.

Forty-one fungal isolates (one isolate per species) representing common plant pathogens and food crop contaminants were grown on sterile, polished rice and assayed for mutagenic activity in the Salmonella typhimurium-microsome system. Initially, single doses of aqueous and chloroform extracts of the moldy rice were assayed against the TA100 tester strain by incorporating extracts into the growth medium and by applying small quantities on disks placed on the agar surface. Suspected activity was examined further by analysis of several doses in the plate incorporation assay. Extracts of two aflatoxin-producing isolates (Aspergillus flavus and A. parasiticus) showed pronounced mutagenic activity, as did extracts of five other isolates (A. heterothallicus, A. nidulans, A. terricola, Alternaria tenuis, and Fusarium moniliforme) which did not contain detectable aflatoxins. Seven additional isolates (Botrytis cineria, Ceratocystis fimbriata, Cladosporium herbarum, Fusarium solani f. sp. pisi, Penicillium oxalicum, Thermomyces lanuginosus, and Verticilium albo-atrum) revealed activity which was possibly mutagenic; i.e., mutagenic responses were not observed in both the disk and incorporation assays, and clear dose-related activity was not observed in the incorporation assay. Extracts of the remaining fungi were not mutagenic in the bacterial assay.     

Colonization of cotton roots by soil micro-organisms

An experiment was carried out to study the colonization of soil microorganisms on cotton roots buried in the soil.Fusarium culmorum, F. solani, F. oxysporum, Macrophomina phaseoli andBacillus sp. were found to be the predominant colonizers followed byBotryodiplodia theobromae, Fusarium acuminatum, Aspergillus niger, A. flavus, Stachybotrys atra, Tilachlidium humicola andRhizopus sp. The practice of leaving cotton roots in the soil would therefore add to the building up of the inoculum potential of the soil inhabiting fungi several of which are serious root pathogens.     

Phytotoxic activity of chernozem saprophytic micromycetes: specificity, sorption and stability of phytotoxins in soil

Micromycetes of the complex of typical chernozem saprotrophic fungi released phytotoxic metabolites into medium. The metabolites displayed their phytotoxic activities directly in soil. Evaluation of the toxicities, range of biological effects activities, and stabilities of phytotoxins in soil and the rates of their biodegradation allowed the species that can serve as indicators of chernozem microbial toxicosis to be selected, namely, Aspergillus clavatus, Fusarium solani, Talaromyces flavus, Penicillium rubrum, and P. funiculosum.     

Detection of mutagens produced by fungi with the Salmonella typhimurium assay.

Forty-one fungal isolates (one isolate per species) representing common plant pathogens and food crop contaminants were grown on sterile, polished rice and assayed for mutagenic activity in the Salmonella typhimurium-microsome system. Initially, single doses of aqueous and chloroform extracts of the moldy rice were assayed against the TA100 tester strain by incorporating extracts into the growth medium and by applying small quantities on disks placed on the agar surface. Suspected activity was examined further by analysis of several doses in the plate incorporation assay. Extracts of two aflatoxin-producing isolates (Aspergillus flavus and A. parasiticus) showed pronounced mutagenic activity, as did extracts of five other isolates (A. heterothallicus, A. nidulans, A. terricola, Alternaria tenuis, and Fusarium moniliforme) which did not contain detectable aflatoxins. Seven additional isolates (Botrytis cineria, Ceratocystis fimbriata, Cladosporium herbarum, Fusarium solani f. sp. pisi, Penicillium oxalicum, Thermomyces lanuginosus, and Verticilium albo-atrum) revealed activity which was possibly mutagenic; i.e., mutagenic responses were not observed in both the disk and incorporation assays, and clear dose-related activity was not observed in the incorporation assay. Extracts of the remaining fungi were not mutagenic in the bacterial assay.     

Antifungal activity of rhizospheric bacteria

Fluorescent Pseudomonad spp. were isolated from the rhizosphere of potato plants (Algeria) by serial dilutions of rhizosphere soils on Kings B medium and were tested for their antifungal activity. The antifungal activity of the Pseudomonas isolated from Potatoes rhizosphere was tested against Pythium ultimum, Rhizoctonia solani and Fusarium oxysporum in dual culture with bacteria on PDA. The Petri dish was divided into tow, on one the bacteria was spread and on the opposite side fungal plugs were inoculated and incubated for one week. Fourteen bacteria were isolated; only one isolate inhibited the growth of Pythium ultimum, Rhizoctonia solani, Fusarium solani; Fusarium oxysporum f.sp. albedinis and Fusarium oxysporum f. sp. Lycopersici with inhibition zones of 39.9, 33.7, 30.8, 19.9 and 22.5 mm respectively.     

Characterization of a fungistatic substance produced by Aspergillus flavus isolated from soil and its significance in nature

A fungus capable of using vegetable tissues for multiplication in soil was isolated and identified as Aspergillus flavus based on morphological characteristics and sequence similarity of ITS and 28S. When grown in liquid medium prepared from the same vegetable tissues used in soil amendment, the isolate of A. flavus produced a substance capable of preventing disease development of black leaf spot of mustard cabbage caused by Alternaria brassicicola and inhibiting the germination of A. brassicicola conidia. The inhibitory substance was fungistatic, and was very stable under high temperature and high or low pH value. It was soluble in ethanol or methanol, moderately soluble in water, and insoluble in acetone, ethyl acetate or ether. The inhibitor is not a protein and has no charges on its molecule. This is the first discovery of the production of a fungistatic substance by this deleterious fungus. Results from this study suggest the possession of a strong competitive saprophytic ability by A. flavus, which in turn may explain the widespread occurrence of this fungus in soils. Production of a fungistatic substance when A. flavus was grown in medium prepared from vegetable tissues suggests the importance of antibiotic production in its competitive saprophytic colonization of organic matters in soils.     

Competition between soil bacteria and fusarium

Summary The phenomenon of competition has been characterized in liquid medium and sterile soil systems using a variety of soil bacteria andFusarium oxysporum f.cubense as test organisms. For most of the bacteria, suppression of the fungus was the result of a biologically induced nitrogen deficiency, this effect being reversed by the addition of excess inorganic nitrogen. High populations of competitors were found in two soils of neutral pH, but no isolates competed in the acid San Alejo loam.Agrobacterium radiobacter was able to compete when San Alejo loam was limed to about pH 6.6. Inhibition of the fungus by a number of gram-positive, spore-forming rods could not be accounted for in terms of competition for nutrients or by antibiotic production in artificial media.The competitive ability ofA. radiobacter when tested in twelve Central American soils was found to be related to pH in acid and neutral environments but was correlated with texture, organic-matter content and total nitrogen in soils of intermediate pH. In all soils where inhibition occurred, the competitive effect was overcome by additions of inorganic nitrogen. Excluding the group ofBacillus spp., the competitive ability of soil bacteria was related to the ability to develop in the absence of amino acids and growth factors but could not be correlated with growth rates of the bacteria in soil or liquid medium.It is suggested that competition for nutrients is a significant means of ecological control among members of the soil microflora and, together with competitive interactions for space and oxygen, may be the major factors governing the biological control of soil-borne fungi.The investigation was supported in part by a grant from the United Fruit Company. Agronomy Paper No. 471.     

Secretion of keratinolytic enzymes and keratinolysis by Scopulariopsis brevicaulis and Trichophyton mentagrophytes: regression analysis

A survey on keratinophilic fungi from poultry-farm soils at Namakkal and from feather dumping soils at Chennai, India, revealed the existence of 34 species of fungi. Most of the fungi exhibited variable efficiency in producing extracellular keratinase when grown in plates with chicken feathers as the sole carbon and nitrogen source. The fungi Aspergillus flavus, Aspergillus niger, Aspergillus versicolor, Chrysosporium state of Arthroderma tuberculatum, Paecilomyces carneus, Scopulariopsis brevicaulis, Trichoderma viride, and Trichophyton mentagrophytes were efficient candidates to degrade the feathers. However, when cultivating the strains in submerged conditions in a medium containing chicken feathers as the sole nutrients source, Aspergillus glaucus, Chrysosporium keratinophilum, Curvularia lunata, Fusarium solani, and Penicillium citrinum also proved to be potent. Among all species, S. brevicaulis and Trichophyton mentagrophytes produced higher amounts of keratinase in both methods. Conditions for keratinase production were optimized by statistical design and surface plots. The highest keratinase activity was estimated by S. brevicaulis (3.2 KU/mL) and Trichophyton mentagrophytes (2.7 KU/mL) in the culture medium with chicken feathers and shows (79% and 72.2% of degrading ability, respectively).     

重金属对土壤中萝卜种子发芽与根伸长抑制的生态毒性

高等植物是生态系统中的基本组成部分。一个平衡、稳定的生态系统生产健康、优良的高等植物。反之 ,一个不稳定或受到外来污染的生态系统 ,对高等植物的生长可带来不利和可见的负面影响。因此 ,利用高等植物的生长状况监测土壤污染程度 ,是从生态学角度衡量土壤健康状况 ,评价土壤质量的重要方法之一[4 ,6] 1) 。土壤生态毒理学评价方法是对化学分析方法的重要补充。目前已建立的高等植物毒理试验有三种方法 ,即 1根伸长试验 ;2种子发芽试验 ;3早期植物幼苗生长试验[3 ,5,6,10 ] 。最初 ,这类试验主要用于纯化学品的毒性检验 ,但随着对土壤…     

Phytotoxic Activities of Chernozem Saprotrophic Micromycetes: Specificity,Sorption, and Stability of Phytotoxins in Soil

Micromycetes of the complex of typical chernozem saprotrophic fungi released phytotoxic metabolites into their growth medium. The metabolites displayed their phytotoxic activities directly in soil. Evaluation of the toxicity, range of biological effects, activity, and stability of the phytotoxins in soil, as well as of the rates of their biodegradation, made it possible to select species that can serve as indicators of microbial toxicosis of chernozem (Aspergillus clavatus, Fusarium solani, Talaromyces flavus, Penicillium rubrum, and P. funiculosum).     

Real-time detection of common microbial volatile organic compounds from medically important fungi by Selected Ion Flow Tube-Mass Spectrometry (SIFT-MS)

We describe a new method, Selected Ion Flow Tube-Mass Spectrometry (SIFT-MS) for the rapid and sensitive real-time detection and quantification of volatile organic compounds from medically important fungi, grown on a range of laboratory media. SIFT-MS utilises the chemical ionisation reactions of mass-selected ions to characterise volatile organic compounds (VOCs) that are produced as metabolites from fungi. This technique has the distinct advantage over others in that it readily detects low molecular weight, reactive volatiles, and allows for real-time, quantitative monitoring. The fungi examined in this study were Aspergillus flavus, Aspergillus fumigatus, Candida albicans, Mucor racemosus, Fusarium solani, and Cryptococcus neoformans grown on or in malt extract agar, Columbia agar, Sabouraud's dextrose agar, blood agar, and brain-heart infusion broth. Common metabolites (ethanol, methanol, acetone, acetaldehyde, methanethiol, and crotonaldehyde) were detected and quantified. We found the fingerprint of volatiles, in terms of presence and quantity of volatiles to be strongly dependent on the culture medium, both in terms of variety and quantity of volatiles produced, but may form the basis for species specific identification of medically important fungi.     

Expression of the Talaromyces flavus glucose oxidase gene in cotton and tobacco reduces fungal infection,but is also phytotoxic

Glucose oxidase secreted by the fungus Talaromyces flavus generates, in the presence of glucose, hydrogen peroxide that is toxic to phytopathogenic fungi responsible for economically important diseases in many crops. A glucose oxidase gene from T. flavus, was modified with a carrot extensin signal peptide and fused to either a constitutive or root-specific plant promoter. T1 tobacco plants expressing the enzyme constitutively were protected against infection by the seedling pathogen Rhizoctonia solani. Constitutive expression in tobacco was associated with reduced root growth, and slow germination on culture medium, and with reduced seed set in glasshouse conditions. Several independent transformed cotton plants with a root-specific construct expressed high glucose oxidase activity in the roots, excluding the root tip. Selected T3 homozygous lines showed some protection against the root pathogen, Verticillium dahliae, but not against Fusarium oxysporum. High levels of glucose oxidase expression in cotton roots were associated with reduced height, seed set and seedling germination and reduced lateral root formation. If this gene is to be of value for crop protection against pathogens it will require precise control of its expression to remove the deleterious phenotypes. This revised version was published online in June 2006 with corrections to the Cover Date.     

FUNGISTASIS IN SOILS

1. Fungistasis in soil is a widespread phenomenon affecting most fungal propagules, though some are insensitive. In most instances, it is coexistent with the presence of living microorganisms, and is annulled by energy-yielding nutrients. Fungistasis with characteristics similar to that in soil may also occur on leaves of plants. 2. Germination and growth of bacteria and actinomycetes is also restricted in soils. The characteristics of their inhibition appear to be the same as those for fungi. Therefore, the concept of a widespread microbial inhibition in soil can be applied to all three groups of microorganisms. 3. Fungistasis can be detected by various direct methods, or indirectly by methods involving the use of porous or permeable carriers. It may be expressed as a restriction on the final amount of germination (the usual parameter), germination rate (with time), and rate of germ-tube or hyphal growth. Since the expression of fungistasis is often complete in soil, titration with nutrients may be required to distinguish between the sensitivities of different fungi. 4. Fungistasis generally is expressed most strongly at soil moisture contents somewhat less than saturation. Its expression usually is maximal in neutral or slightly alkaline soils. In acidic conditions fungistasis may be lessened because of suppression of bacterial and actinomycete activity. Increased sensitivity of some fungi in soils of pH > 7.0 may be caused by a directly unfavourable effect of pH on the fungus. 5. Fungal species with small spores tend to be highly sensitive to fungistasis. These spores tend to germinate slowly and to require exogenous nutrients for germination. By contrast, species with larger spores and sclerotia often do not require exogenous nutrients for germination. The larger spores tend to germinate rapidly and to exhibit low sensitivity, as compared with small spores. A few nutrient-independent spores are insensitive to fungistasis. At least a part of the difference in sensitivity is related to germination time; spores which germinate slowly compete poorly with the soil micro-flora for their nutrients. 6. Fungistasis is often temporarily annulled by enriching the soil with energy-yielding nutrients. Usually, complex materials such as plant residues are most effective. A few weeks after such treatment, the level of fungistasis may, however, be increased. Annulment of fungistasis by compounds not utilized as energy sources has not yet been demonstrated. 7. Several soils naturally suppressive to Fusarium wilt diseases were more fungistatic to Fusarium than soils conducive to wilt. Potential means by which fungistasis may be manipulated to control root-infecting fungi are (a) through stimulation of germination with nutrients, thus exposing the germ tube to lysis, and (b) by increasing the fungistatic level of soil through appropriate amendments. 8. Volatile substances identified in soils, some of which are potentially inhibitory to fungi include (a) ammonia, which apparently is evolved from ammonium salts in some arid soils of high pH, (b) ethylene, which has been identified in some soils of pH < 7.0 (though high levels of this gas seem to be tolerated by most fungi), (c) allyl alcohol, and (d) other unidentified substances. Non-volatile inhibitors include high molecular weight substances revealed by molecular sieve chromatography of soil extracts. Microbial metabolites such as those present in staled fungal cultures also have been proposed to account for fungistasis. In a few soils fungistasis persists after sterilization because of the presence of inhibitory concentrations of calcium carbonate, iron or aluminium. Inherent in the proposition that inhibitory substances provide the primary mechanism of fungistasis is the concept of a highly complex phenomenon, involving various highly specific inhibitory and counteracting stimulatory substances, with the outcome for the fungus depending on the kinds and relative amounts of each present. 9. By the nutrient-deficiency hypothesis, the level of available nutrients in soil is insufficient to support germination of nutrient-dependent propagules, except in nutrient-rich microsites. Inhibition of nutrient-independent propagules is explained by loss of endogenous nutrients required for germination, through microbial nutrient competition. Evidence for this hypothesis is (a) the imposition of fungistasis on numerous nutrient-independent propagules during incubation on leaching model systems designed to simulate microbial nutrient competition in soil, (b) similar losses of endogenous nutrients occurring on soil and the leaching system, and (c) the fact that soils are chronically deficient in energy in relation to the microbial populations present, with the consequence that enforced inactivity is imposed upon most of the population at any given time for this reason alone, regardless of the presence or absence of fungistatic substances. Journal series article no. 7747 from the Michigan Agricultural Experiment Station.