Soil Fungistasis Research Articles

SOIL FUNGISTASIS AGAINST FUSARIUM GRAMINEARUM UNDER DIFFERENT CROP MANAGEMENT SYSTEMS

Soil management, in terms of tillage and cropping systems, strongly influences the biological properties of soil involved in the suppression of plant diseases. Fungistasis mediated by soil microbiota is an important component of disease-suppressive soils. We evaluated the influence of different management systems on fungistasis against Fusarium graminearum, the relationship of fungistasis to the bacterial profile of the soil, and the possible mechanisms involved in this process. Samples were taken from a long-term experiment set up in a Paleudult soil under conventional tillage or no-tillage management and three cropping systems: black oat (Avena strigose L.) + vetch (Vicia sativa L.)/maize (Zea mays L.) + cowpea (Vigna sinensis L.), black oat/maize, and vetch/maize. Soil fungistasis was evaluated in terms of reduction of radial growth of F. graminearum, and bacterial diversity was assessed using ribosomal intergenic spacer analysis (RISA). A total of 120 bacterial isolates were obtained and evaluated for antibiosis, and production of volatile compounds and siderophores. No-tillage soil samples showed the highest level of F. graminearum fungistasis by sharply reducing the development of this pathogen. Of the cropping systems tested, the vetch + black oat/maize + cowpea system showed the highest fungistasis and the oat/maize system showed the lowest. The management system also affected the genetic profile of the bacteria isolated, with the systems from fungistatic soils showing greater similarity. Although there was no clear relationship between soil management and the characteristics of the bacterial isolates, we may conclude that antibiosis and the production of siderophores were the main mechanisms accounting for fungistasis.

Assessment of germination and carnivorous activities of a nematode-trapping fungus Arthrobotrys dactyloides in fungistatic and fungicidal soil environment

Carnivorism is the ability of nematode-trapping fungi to trap and digest the nematodes by sophisticated devices called traps. Delivery of nematode-trapping fungi in soil for bio-control of pest nematodes often fails or gives inconsistent results. Possible reasons for failure could be the effect of soil fungistasis on germination of nematode-trapping fungi in soil environment, use of avirulent species and sensitivity of these fungi to fungicidal residues in soil. Exploitation of nematode-trapping fungi for nematode control demands that it be compatible with fungicides applied in soil or crops and proliferate in soil. This investigation represents is one of the first to evaluate the effect of fungicides on the nematode-trapping fungus Arthrobotrys dactyloides. A. dactyloides showed in vitro carnivorous potential against Meloidogyne incognita, Meloidogyne javanica, Meloidogyne graminicola, Helicotylenchus dihystera and Heterodera cajani. Conidia of A. dactyloides exposed to agricultural soils showed poor germination but formed conidial traps, which captured and killed the soil nematodes. Conidial traps, which trapped the nematodes, grew well in all soils after killing and nutrient absorption from nematode body. Soil amended with 20mg aikg−1 of carbendazim and thiram, 30mg aikg−1 of mancozeb, 50mg aikg−1 of captan, and 100mg aikg−1 of carboxin completely checked the conidial trap formation and nematode capturing. 30, 50 and 100mg aikg−1 of metalaxyl adversely affected the conidial trap formation and nematode capturing in soil. Propiconazole inhibited 15.2% conidial trap formation up to 50mg aikg−1 but caused 93.3% inhibition of conidial traps formation and complete inhibition of nematode capturing at 100mg aikg−1. Sulphur, triademefon, and tricyclazole showed least toxic effect on conidial trap formation and nematode capturing activities of A. dactyloides in soil up to 100mg aikg−1.

Badania właściwości fungistatycznych gleby

This paper presents the results of study the effect of rhizosphere of <i>Pennisetum typhoides</i> on the soil fungistasis. The fungistasis of three different regions of the root has been investigated in relation to rhizosphere effect.

Biocontrol potential of Microsphaeropsis ochracea on microsclerotia of Verticillium longisporum in environments differing in microbial complexity

The potential of the fungal antagonist Microsphaeropsis ochracea to control the soilborne pathogen Verticillium longisporum was investigated in environments with varying microbial complexity (in vitro vs. in vivo, sterile vs. unsterile, controlled conditions vs. field). A semi-quantitative PCR assay was developed for the detection of M. ochracea on unsterile plant debris. In vitro, M. ochracea caused high levels of mortality to V. longisporum microsclerotia (51–100 %) from 4 to 24 °C, with a broad optimum between 16 and 24 °C. In controlled conditions, M. ochracea significantly reduced the viability of V. longisporum microsclerotia grown on dead rapeseed stems in autoclaved sand, but not in unsterile soil. Likewise, in two experimental years, no significant reduction of V. longisporum inoculum was detectable on rapeseed straw buried in small plots in the field in any of the treatments (soil depths, exposure duration, doses of M. ochracea). Germination of M. ochracea pycnidiospores was inhibited by general soil fungistasis in unsterile soil from a field, botanical garden and grassland. Accordingly, V. longisporum infection of rapeseed plants in the greenhouse was reduced only at artificially high doses of M. ochracea inoculum and no biocontrol efficacy in disease control was recorded in field experiments conducted with winter oilseed rape during two subsequent seasons in an experimental field near Gottingen, with a soil homogenously infested with V. longisporum. The results demonstrate that M. ochracea, although having shown promising potential in controlling pathogens with melanised resting structures on leaf litter, evidently lacks microbial competitiveness to effectively control pathogens in the soil such as V. longisporum, even though the latter is effectively inhibited in vitro.

Fungal diversity increases soil fungistasis and resistance to microbial invasion by a non resident species

Biodiversity decline is a major concern for ecosystem functioning. Recent research efforts have been mostly focused on terrestrial plants, while, despite their importance in both natural and artificial ecosystems, little is known about soil microbial communities. This work aims at investigating the effects of fungal species richness on soil invasion by non resident microbes. Synthetic fungal communities with a species diversity ranging from 1 to 8 were assembled in laboratory microcosms and used in three factorial experiments to assess the effect of diversity on soil fungistasis, microbial invasion of soil amended with plant litter and of plant rhizosphere. The capability of different microbes to colonize environments characterized by different resident microbial communities was measured. The number of microbial species in the microcosms positively affected soil fungistasis that was also induced more rapidly in presence of synthetic communities with more species. Moreover, the increase of resident fungal diversity dramatically reduced the invasibility of both soil and plant rhizosphere. We found lower variability of soil fungistasis and invasibility in microcosms with higher species richness of microbial communities. Our study pointed out the existence of negative relationships between fungal diversity and soil invasibility by non resident microbes. Therefore, the loss of microbial species may adversely affect ecosystem functionality under specific environmental conditions.

Biochemical quality of organic amendments affects soil fungistasis

Soilborne plant pathogens are among the most important limiting factors for the productivity of agro-ecosystems. Identifying reliable and effective control methods is crucial for efficient biological control. Soil fungistasis is the capability of soils to inhibit the germination and growth of soil-borne fungi in presence of optimal abiotic conditions. The aim of this study is to clarify the relationships between soil amendments with plant residues spanning a wide variety of biochemical quality and soil fungistasis. Microcosms experiments were performed with 42 different plant residues and the effect on soil fungistasis was assessed by using four different fungi (Aspergillus niger, Botrytis cinerea, Pyrenochaeta lycoperici and Trichoderma harzianum). We measured soil respiration and FDA enzymatic activity and compared classic litter proximate chemical analysis with 13C-CPMAS NMR spectroscopy. Results showed that quality of organic amendments is a major controlling factor of soil fungistasis. The dramatic relief of soil fungistasis when soil was amended with lignin poor, but labile C rich, substrates gives strong support to the competition-based hypothesis. The positive correlation between soil respiration and fungal growth further supports the competition hypothesis. Finally, 13C NMR results showed a relationship between soil fungistasis and the biochemical quality of plant residues, and provided a quantitative assessment of the time required for fungistasis restoration after organic materials application.

Germination of Lecanicillium fungicola in the mycosphere of Agaricus bisporus

Dry bubble disease is a major problem in the commercial cultivation of the white button mushroom Agaricus bisporus and is caused by the ascomycete Lecanicillium fungicola. In the casing layer, germination of L. fungicola spores is inhibited by the microflora, a phenomenon known as fungistasis. Fungistasis is annulled when the casing is colonized by A. bisporus hyphae. We demonstrated that addition of A. bisporus-associated sugars, similarly annulled the casing fungistasis. However, casing fungistasis does not seem to be based on competition for resources as L. fungicola spores germinate regardless of nutrient availability. Pseudomonas bacteria are a dominant group of bacteria in the casing and have previously been implied to be essential for the development of fungistasis in soils. Antibiotics produced by model strain Pseudomonas fluorescens CHA0 inhibited L. fungicola spore germination. Addition of glucose desensitized spores of L. fungicola, which resulted in germination in the presence of antibiotics. We conclude that antibiotics produced by the microflora most likely cause fungistasis.

Management of Root Knot Disease in Rice Caused by Meloidogyne graminicola through Nematophagous Fungi

A glass house experiment was conducted for the effectiveness of nematophagous fungi against Meloidogynegraminicola in which Arthrobotrys oligospora and Dactylaria eudermata were used for the management of rootknot disease of rice. Root knot nematodes, Meloidogyne graminicola had proved itself as an important limitingfactor for successful cultivation and productivity of rice. By amending the soil with application of mass cultureof two nematophagous fungi (Arthrobotrys oligospora and Dactylaria eudermata) reduced the number of rootgalls by 86.9% and 81.1%, of females by 94.2% and 91.7%. The mass culture of these fungi increased the plantgrowth: shoot length by 41.9% and 38.8%, root length by 44.6% and 41.8%, fresh weight of shoot by 61.1% and58.7%, and fresh weight of root by 24.3% and 22.5%, respectively over nematode infested soil. The betterperformance of A. oligospora may be attributed to better colonization and establishment of A. oligospora than D.eudermata and may be better tolerance of the fungus to soil fungistasis.

Fungistatic Intensity of Agricultural Soil Against Fungal Agents and Phylogenetic Analysis on the Actinobacteria Involved

A total of 287 agricultural soil samples collected from 26 provinces or autonomous regions of China were tested on their ability to suppress the conidial germination of nine biocontrol fungal agents. These soil samples showed great differences in the degree to inhibit the germination of conidia (22.8% < mean inhibition rate < 97.5%), but all exhibited fungistatic activities above the moderate levels (mean inhibition rate > 50%) to most of tested fungi. Ten soil samples that have stronger fungistatic intensity (germination inhibition rate > 68.3%) to the target fungi, Trichoderma viride and Paecilomyces lilacinus, were selected to evaluate their soil actinobacteria involved fungistasis in soil. Of the 1,000 isolates from those soil samples, 345 actinobacteria exhibited fungistatic activity to conidial germination of T. viride and P. lilacinus with germination inhibition rates higher than 10%. Sequences encoding 16S rRNA gene of the 345 actinobacteria were analyzed by ARDRA and resulted 44 different ARDRA types. Fifty-six isolates, at least one from each unique ARDRA type, were selected for 16S rDNA sequencing and phylogenetic analysis. Results indicated that the actinobacteria involved in the soil fungistasis had close phylogenetic relationship with the members of Sterptomycetaceae, Microbacteriaceae, Micrococcaceae, and Nocardiacea.

Fungistasis and general soil biostasis – A new synthesis

In most soils, fungal propagules are restricted to a certain extent in their ability to grow or germinate. This phenomenon, known as soil fungistasis, has received considerable attention for more than five decades, mostly due to its association with the general suppression of soil-borne fungal diseases. Here, we review major breakthroughs in understanding the mechanisms of fungistasis. Integration of older fungistasis research and more recent findings from different biological and chemical disciplines has lead to the consensus opinion that fungistasis is most likely caused by a combination of microbial activities, namely withdrawal of nutrients from fungal propagules and production of fungistatic compounds. In addition, recent findings indicate that there are mechanistic links between these activities leading towards an integrated theory of fungistasis. Among the potentially fungistatic compounds volatiles have received particular attention. Whereas it has long been assumed that fungistasis is the result of the metabolic activity of the total soil microbial biomass, more recent research points at the importance of activities of specific components of the microbial community. These insights into fungistasis have also formed the basis for strategies to increase general soil suppression. Besides these basic and practical aspects of fungistasis, its impact on fungal ecology, in particular on fungal exploration strategies, is discussed. Finally, we take a closer look at plant–soil feedback experiments to demonstrate the occurrence of fungistasis-like phenomena and to suggest that fungistasis may be part of a much wider phenomenon: general soil biostasis.

Development of artificial conidia for ecological studies of Rhizoctonia solani in soil

Artificial conidia of Rhizoctonia solani were developed by releasing protoplasts from young mycelia with lytic enzymes and by inducing cell wall formation in stabilizer solution. Conidia produced in this way were spherical with sizes ranging from 10 to 20μm in diameter. Artificial conidia were sensitive to soil fungistasis. Young hyphae originated from artificial conidia were also sensitive to fungistasis and mycolysis in soils. These results demonstrate that the previously reported insensitivity of R. solani to fungistasis and mycolysis in soils is due to special ability of propagules used rather than the inherited nature of the organism. Germination rates of artificial conidia on soils were inversely correlated with the amount of fungicide Flutolanil added. When germination of artificial conidia was used to detect suppressive soils, 3 out of 30 soil samples collected from different parts of Taiwan were suppressive to R. solani and all these suppressive soils were low in pH. Using artificial conidia for assay of fungicide activity in soil and detection of suppressive soils has the advantages of being fast and precise in comparison with relative hyphal growth. However, preparation of artificial conidia at this stage is tedious and time-consuming.

Volatiles produced by interacting microorganisms potentially useful for the control of plant pathogens

The results of studies about interactions between microorganisms involving at least one plant pathogen are of interest to the areas of ethiology and control in Plant Pathology. Various aspects of these interactions have been studied over the years but the toxicity of volatile organic compounds (VOCs) has been emphasized only recently, developing techniques and procedures, and producing additional knowledge to those already obtained with water-soluble substances. This new facet of these interactions based on VOCs is discussed in this review involving mainly fungi, bacteria and nematodes pathogenic to plants. Also discussed is the role of VOCs produced by microorganisms, especially fungi and bacteria, in soil fungistasis and the effect of VOCs on fungal agents used in the control of plant parasitic nematodes. The evidence of VOCs broadens the research studies about these interactions. However, the scarcity of the research results in this area show up gaps which need to be filled and some research proposals are discussed. The present and future accumulated VOC knowledge will perhaps be beneficial to farmers, especially aspects related to increasing soil suppressiveness to plant disease and to the finding of analog molecules of VOCs highly effective against plant pathogens.

Soil fungistasis and its relations to soil microbial composition and diversity: A case study of a series of soils with different fungistasis

Fungistasis is one of the important approaches to control soil-borne plant pathogens. Some hypotheses about the mechanisms for soil fungistasis had been established, which mainly focused on the soil bacterial community composition, structure, diversity as well as function. In this study, the bacterial community composition and diversity of a series of soils treated by autoclaving, which coming from the same original soil sample and showing gradient fungistasis to the target soil-borne pathogen fungi Fusarium graminearum, was investigated by soil bacterial 16S rDNA-PCR (polymerase chain reaction) cloning, restriction fragment length polymorphism (RFLP), and sequencing. The results showed that the soil fungistasis capacity was closely correlated with soil bacterial community composition and diversity, such as soil fungistasis declined with the decrease of soil bacterial diversity. Meanwhile, the bacterial community composition and structure were significantly different along the gradient of soil fungistasis tested. alpha-Proteobacteria, beta-Proteobacteria, Flexibacter, and some uncultured soil bacteria were contributed to soil fungistasis in combination with some other special bacteria (Pseudomonas and Acidobacteria) which were know to be key species in suppression of fungal growth.

Possible contributions of volatile-producing bacteria to soil fungistasis

Soil fungistasis can adversely affect the germination and growth of most fungal species in the field. Among the inhibitors, volatiles of microbial origins are potentially very important. In this study, we investigated the frequency and identity of bacteria producing fungistatic volatiles. Among the 1018 bacterial isolates tested, 328 were found to produce antifungal volatiles that could inhibit spore germination and mycelial growth of two nematicidal fungi Paecilomyces lilacinus and Pochonia chlamydosporia. A phylogenetic analysis based on restriction fragment length polymorphism (RFLP) and 16S rDNA sequence placed the 328 bacteria in five groups: Alcaligenaceae, Bacillales, Micrococcaceae, Rhizobiaceae and Xanthomonadaceae. Volatile compounds of 39 bacterial isolates were identified by gas chromatography/mass spectrum (GC/MS). Tests with commercially available antifungal compounds suggested that seven volatile compounds of bacterial origins (acetamide, benzaldehyde, benzothiazole, 1-butanamine, methanamine, phenylacetaldehyde and 1-decene) likely play important roles in soil fungistasis.

Compounds inhibitory to nematophagous fungi produced by Bacillus sp. strain H6 isolated from fungistatic soil

Soil fungistasis can cause inconsistent control by nematophagous fungi of plant-pathogenic nematodes in field situations. Recent studies have shown that production of fungistatic compounds by bacteria was the principal explanation for soil fungistasis. The culture filtrate of Bacillus sp. strain H6, a strain representative of the dominant colony types isolated from fungistatic soils, showed strong inhibitory activity against nematophagous fungi by inducing unusual swelling in the conidia and the germ tubes of nematophagous fungi, thereby preventing the fungi from proliferating. This inhibitory mechanism is novel in comparison with other known mechanisms. Antifungal activity of the culture filtrate of strain H6 was maximal after culture in Luria Bertani (LB) broth (pH 7.0) at 36°C for 36 h. The inhibitory effect of the compounds produced by Bacillus sp. strain H6 was not significantly influenced by pH, and the inhibitory compounds in the culture filtrate were thermostable. After being partially purified by thin layer chromatography (TLC) on silica gel plates, characterization by colour reactions and positive fast atom bombardment mass spectrometry indicated that the inhibitory compounds showed similarity to iturin A group compounds.

Screening of fungal strains responsible for strong fungistasis against Fusarium oxysporum f. sp. radicis-lycopersici in a coffee compost-amended soil

(pp. 817–824) Disease incidence of crown and root rot of tomato, caused by Fusarium oxysporum f. sp. radicis-lycopersici J3 (FOL J3) was significantly lower in a soil (CC-soil) amended with coffee compost and chemical fertilizers (CF) than in a soil (CF-soil) amended with only CF. Germination of microconidia of various plant-pathogenic Fusarium oxysporum strains was consistently lower in the CC-soil than in the CF-soil, suggesting that the CC-soil possessed a higher degree of fungistasis. When the CC-soil was supplemented with rifampicin and kanamycin, germination of FOL J3 didn't increase, suggesting that the higher degree of fungistasis in the CC-soil may not be of bacterial origin. The substrate-induced respiration inhibition method demonstrated that the CC-soil possessed higher microbial activity and was dominated by fungi. PCR-DGGE analysis showed that the microbial community structure of the two soils was different. Fungal mycelia were isolated from the soils and the effect of the isolates on soil fungistasis was examined. Three isolates, all belonging to F. oxysporum out of 49 showed strongest suppressive effect on the germination of FOL J3 and two isolates suppressed Fusarium crown and root rot disease when they were inoculated into autoclaved CC-soil. These results might suggest that the isolates close to F. oxysporum were responsible for highly fungistatic capability in the CC-soil and were a possible source for disease suppression.

Soil volatile fungistasis and volatile fungistatic compounds

Fungistasis is a widespread phenomenon that can be mediated by soil microorganisms and volatile organic compounds (VOCs). The relationship between soil microorganisms and VOCs is still unclear, however, and many fungistatic compounds remain to be identified. We assessed the effects of soils (soil direct fungistasis) and VOCs produced by natural soils (soil volatile fungistasis) on the spore germination of several fungi. Both strong soil direct fungistasis and soil volatile fungistasis were observed in a wide range of soils. Soil fungistasis and VOC fungistasis were significantly correlated ( P<0.001). The volatile fungistatic activity of soils stopped after autoclaving. Some VOCs were identified by using solid-phase microextraction-gas chromatography/mass spectrum. VOC composition and in vitro antagonism of relatively pure commercial compounds also were measured. Some VOCs, trimethylamine, 3-methyl-2-pentanone, dimethyl disulfide, methyl pyrazine, 2,5-dimethyl-pyrazine, benzaldehyde, N, N-dimethyloctylamine and nonadecane, were produced by various fungistatic soils. Moreover, antifungal activity test of above VOCs showed that trimethylamine, benzaldehyde, and N, N-dimethyloctylamine have strong antifungal activity even at low levels (4–12 mg l −1).

Microbial community composition affects soil fungistasis.

Most soils inhibit fungal germination and growth to a certain extent, a phenomenon known as soil fungistasis. Previous observations have implicated microorganisms as the causal agents of fungistasis, with their action mediated either by available carbon limitation (nutrient deprivation hypothesis) or production of antifungal compounds (antibiosis hypothesis). To obtain evidence for either of these hypotheses, we measured soil respiration and microbial numbers (as indicators of nutrient stress) and bacterial community composition (as an indicator of potential differences in the composition of antifungal components) during the development of fungistasis. This was done for two fungistatic dune soils in which fungistasis was initially fully or partly relieved by partial sterilization treatment or nutrient addition. Fungistasis development was measured as restriction of the ability of the fungi Chaetomium globosum, Fusarium culmorum, Fusarium oxysporum, and Trichoderma harzianum to colonize soils. Fungistasis did not always reappear after soil treatments despite intense competition for carbon, suggesting that microbial community composition is important in the development of fungistasis. Both microbial community analysis and in vitro antagonism tests indicated that the presence of pseudomonads might be essential for the development of fungistasis. Overall, the results lend support to the antibiosis hypothesis.

Influence of pH and Matric Potential on Germination of Cephalosporium gramineum Conidia

Germination of Cephalosporium gramineum conidia in soil was up to twofold greater at -0.064 MPa than at -0.037 and -0.007 MPa when incubated at 5°C for 2 days. Soil pH from 4.7 to 7.5 did not have a significant influence on germination of conidia and the interaction between soil pH and matric potential on germination was not significant. Soil fungistasis, which was previously observed for conidia of C. gramineum, was not observed in these studies. Germination of conidia on mineral salts agar containing phosphate buffer was significantly less at pH 4.5 than at 5.5, 6.5, or 7.5 at 5°C in one of two experiments; however, pH had no influence on germination at 10 or 20°C in two experiments. Although Cephalosporium stripe is more severe under conditions of high soil moisture and low soil pH, increased germination of conidia in response to these factors does not explain the observed increase in disease.

Effect of Soil pH and Soil Type on the Germinability of Phialospores of Gliocladium virens Millers, Giddens and Foster

Phialospores of four isolates of Gliocladium virens viz., 15GV 1 , 26GV 1 , 1AGV 2 and IAGV 6 responded differentially to soil fungistasis in different soils of West Bengal according to the properties of soils explored. The pH of soil appeared to be the cardinal factor. The effect of soil fungistasis was more pronounced in alkaline soil followed by neutral soil but gradually decreased due to reduction of soil pH to acidic level. Fungistasis was more in unsterilized soil compared to sterilised one.