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Biological control of soil-borne pathogens by fluorescent pseudomonads.

Particular bacterial strains in certain natural environments prevent infectious diseases of plant roots. How these bacteria achieve this protection from pathogenic fungi has been analysed in detail in biocontrol strains of fluorescent pseudomonads. During root colonization, these bacteria produce antifungal antibiotics, elicit induced systemic resistance in the host plant or interfere specifically with fungal pathogenicity factors. Before engaging in these activities, biocontrol bacteria go through several regulatory processes at the transcriptional and post-transcriptional levels.

Anti-Bacterial Agents↗

Biological control of fungal pathogens.

Biological control of soil-borne plant pathogens is a potential alternative to the use of chemical pesticides, which have already been proved to be harmful to the environment. Several strains of the fungus Trichoderma have been isolated and found to be effective biocontrol agents of various soil-borne plant pathogenic fungi under greenhouse and field conditions. Different application approaches have been used including integration of Trichoderma with reduced doses of chemical agents. Biochemical and molecular biology studies carried out to explore the mechanisms involved in biological control revealed that Trichoderma is a rather specific mycoparasite. Lectins were found to be involved in the recognition between Trichoderma and its host fungi, whereas chitinase is involved in the degradation of the host cell wall. Genetic engineering techniques were employed in order to increase the effectiveness, stability, and biocontrol capacity of Trichoderma spp. as well as other biocontrol agents, such as Pseudomonass spp. and Rhizobium.

Chitinases↗

Rhizobia as a biological control agent against soil borne plant pathogenic fungi.

Rhizobia promote the growth of plants either directly through N2 fixation, supply of nutrients, synthesis of phytohormones and solubilization of minerals, or indirectly as a biocontrol agent by inhibiting the growth of pathogens. The biocontrol effect of rhizobia is due to the secretion of secondary metabolites such as antibiotics and HCN. Siderophore production in iron stress conditions provides rhizobia an added advantage, resulting in exclusion of pathogens due to iron starvation.

Fungi↗

Kenneth Frank Baker--pioneer leader in plant pathology.

Kenneth F. Baker (1908-1996) made major contributions to understanding diseases of ornamental plants, seed pathology, soil-borne plant pathogens, biological control, and history of plant pathology. His work set the stage for the success of today's ornamentals and nursery industries. His leadership and writings created the scientific framework for research and teaching on soil-borne plant pathogens and biological control. After B.Sc. and Ph.D. degrees from Washington State University in 1930 and 1934, respectively, and one year as a National Research Council Fellow with B.M. Dugger at Wisconsin, he took jobs in 1935 with the U.S. Department of Agriculture in Nebraska on establishment of shelter belts and 1936-39 with the Pineapple Producers Cooperative Association in Hawaii. He worked on diseases of ornamental plants at the University of California, Los Angeles, starting in 1939, moving to Berkeley in 1961 when the UCLA program closed. He retired in 1975 and moved to Corvallis, OR, as Emeritus Professor, Oregon State University, and Collaborator, U.S. Department of Agriculture, Agricultural Research Service. He spent four sabbatical leaves in Australia, and was elected fellow of the American Association for the Advancement of Science in 1950, fellow of the American Phytopathological Society in 1969, and the Horticultural Hall of Fame in 1976.

Botany↗

Isolation and characterisation of a partial peptide synthetase gene from Trichoderma asperellum.

Many species of Trichoderma have attracted interest as agents for the biological control of soil borne fungal pathogens of a range of crop plants. Research on the biochemical mechanisms associated with this application has focused on the ability of these fungi to produce enzymes which lyse fungal cell walls, and antifungal antibiotics. An important group of the latter are the non-ribosomal peptides called peptaibols. In this study Trichoderma asperellum, a strain used in biological control in Malaysia, was found to produce the peptaibol, trichotoxin. This type of peptide molecule is synthesised by a peptide synthetase (PES) enzyme template encoded by a peptide synthetase (pes) gene. Using nucleotide sequences amplified from adenylation (A-) domains as probes, to hybridise against a lambda FIXII genomic library from T. asperellum, 25 clones were recovered. These were subsequently identified as representative of four groups based on their encoding properties for specific amino acid incorporation modules in a PES. This was based on analysis of their amino acid sequences which showed up to 86% identity to other PESs including TEX 1.

Antimicrobial Cationic Peptides↗

Genome sequencing and annotation of two rhizobacteria with antifungal activity: Pseudomonas tolaasii strain A46 and Pseudomonas palleroriana P61.

Here, we report the draft genome sequences of P. tolaasii A46 and P. palleroriana P61, two rhizobacteria previously shown to inhibit Rhizoctonia solani. These genomic resources will support future efforts to elucidate the molecular basis of fungal suppression and to assess the biocontrol potential of these Pseudomonas strains.

antagonistic rhizobacteria↗

Indigenous microflora responses to introduction of cyanogenic strains of Pseudomonas fluorescens into soil.

The effects of cyanogenic Pseudomonas fluorescens strains introduced into soil on the kinetic of colony formation and bacterial community structure were investigated. About 7.8 x 10(8) and 1.2 x 10(9) cfu per g dry soil of TA1 and B2 were added to the soil portions, respectively. The parameters of colony formation by heterotrophic soil bacteria were determined. The bacterial community structure and phenotypic diversity were studied using concept of r/K strategies and echophysiological index, respectively. The physiological state of indigenous heterotrophic bacteria and gram-negative group did not change under the influence of the cyanogenic strains introduced. Phenotypic diversity of the soil bacteria also did not change significantly. However, some short-term shifts in community structure of indigenous heterotrophic bacteria were noticed. This study shows that the introduction of great numbers of cyanogenic P. fluorescens strains could be safely used as potential agents in biological control of soil-born pathogens.

Bacteria↗

Biological control of plant root pathogens.

Rhizobacteria introduced to control soil-borne root diseases must establish metabolically active populations that mediate protection either by direct antagonism of pathogens or by stimulation of host plant defenses. Recent interest has focused on the genetic and biochemical basis of disease control and the influence of environmental factors on the expression and activity of biocontrol mechanisms. The cloning and sequencing of genes involved in the production of microbial metabolites playing key roles in plant defense opens new possibilities for improving the performance of biocontrol agents.

Anti-Bacterial Agents↗

An immunological approach to quantifying the saprotrophic growth dynamics of Trichoderma species during antagonistic interactions with Rhizoctonia solani in a soil-less mix.

Studies of the saprotrophic growth dynamics of Trichoderma species and their fungal hosts during antagonistic interactions are severely hampered by the absence of methods that allow the unambiguous identification and quantification of individual genera in complex environments such as soil or compost containing mixed populations of fungi. Furthermore, methods are required that allow discrimination between active hyphal growth and other components of fungal biomass such as quiescent spores that are produced in large numbers by Trichoderma species. This study details the use of monoclonal antibodies to quantify the saprotrophic growth dynamics of the soil-borne plant pathogen Rhizoctonia solani and biological control strains of Trichoderma asperellum and Trichoderma harzianum during antagonistic interactions in peat-based microcosms. Quantification was based on the immunological detection of constitutive, extracellular antigens that are secreted from the growing tip of Rhizoctonia and Trichoderma mycelium and, in the case of Trichoderma harzianum, from quiescent phialoconidia also. The Trichoderma-specific monoclonal antibody (MF2) binds to a protein epitope of the enzyme glucoamylase, which was shown by immunofluorescence and immunogold electron gold microscopy studies of Trichoderma virens in vitro to be produced at the origin of germ tube emergence in phialoconidia and from the growing tip of germ tubes. In addition, a non-destructive immunoblotting technique showed that the enzyme was secreted during active growth of Trichoderma asperellum mycelium in peat. The Rhizoctonia solani-specific monoclonal antibody (EH2) similarly binds to a protein epitope of a glycoprotein that is secreted during active mycelial growth. Extracts derived from lyophilized mycelium were used as a quantifiable and repeatable source of antigens for construction of calibration curves. These curves were used to convert the absorbance values obtained in ELISA tests of peat extracts to biomass equivalents, which allowed comparisons of the saprotrophic growth dynamics of the pathogen and antagonists to be made in single or mixed species microcosms. Trichoderma species were able to compete successfully with R. solani for nutrients and to prevent saprotrophic growth of the pathogen. Specificity of the Trichoderma quantitative assay was tested in non-sterile soil-based microcosms artificially inoculated with T. asperellum. The assay was highly specific and only detected T. asperellum population dynamics. No cross-reactivity was found with extracts from soil samples containing contaminant fungi.

Animals↗

Mycorrhizosphere interactions to improve plant fitness and soil quality.

Arbuscular mycoruhizal fungi are key components of soil microbiota and obviously interact with other microorganisms in the rhizosphere, i.e. the zone of influence of plant roots on microbial populations and other soil constituents. Mycorrhiza formation changes several aspects of plant physiology and some nutritional and physical properties of the rhizospheric soil. These effects modify the colonization patterns of the root or mycovrhizas (mycorrhizosphere) by soil microorganisms. The rhizosphere of mycorrhizal plants, in practice a mycorrhizosphere, harbors a great array of microbial activities responsible for several key ecosystem processes. This paper summarizes the main conceptual principles and accepted statements on the microbial interactions between mycorrhizal fungi and other members of rhizosphere microbiota and discusses current developments and future trends concerning the following topics: (i) effect of soil microorganisms on mycorrhiza formation; (ii) mycorrhizosphere establishment; (iii) interactions involved in nutrient cycling and plant growth; (iv) interactions involved in the biological control of plant pathogens; and (v) interactions to improve soil quality. The main conclusion is that microbial interactions in the rhizosphere of mycorrhizal plants improve plant fitness and soil quality, critical issues for a sustainable agricultural development and ecosystem functioning.

Bacteria↗

Genetic diversity and biological control activity of novel species of closely related pseudomonads isolated from wheat field soils in South Australia.

Rhizobacteria closely related to two recently described species of pseudomonads, Pseudomonas brassicacearum and Pseudomonas thivervalensis, were isolated from two geographically distinct wheat field soils in South Australia. Isolation was undertaken by either selective plating or immunotrapping utilizing a polyclonal antibody raised against P. brassicacearum. A subset of 42 isolates were characterized by amplified 16S ribosomal DNA restriction analysis (ARDRA), BIOLOG analysis, and gas chromatography-fatty acid methyl ester (GC-FAME) analysis and separated into closely related phenetic groups. More than 75% of isolates tested by ARDRA were found to have >95% similarity to either Pseudomonas corrugata or P. brassicacearum-P. thivervalensis type strains, and all isolates had >90% similarity to either type strain. BIOLOG and GC-FAME clustering showed a >70% match to ARDRA profiles. Strains representing different ARDRA groups were tested in two soil types for biological control activity against the soilborne plant pathogen Gaeumannomyces graminis var. tritici, the causative agent of take-all of wheat and barley. Three isolates out of 11 significantly reduced take-all-induced root lesions on wheat plants grown in a red-brown earth soil. Only one strain, K208, was consistent in reducing disease symptoms in both the acidic red-brown earth and a calcareous sandy loam. Results from this study indicate that P. brassicacearum and P. thivervalensis are present in Australian soils and that a level of genetic diversity exists within these two novel species but that this diversity does not appear to be related to geographic distribution. The result of the glasshouse pot trial suggests that some isolates of these species may have potential as biological control agents for plant disease.

Bacterial Typing Techniques↗

BIOCONTROL WITHIN THE CONTEXT OF SOIL MICROBIAL COMMUNITIES: A Substrate-Dependent Phenomenon.

Broad spectrum biological control of diseases caused by soilborne plant pathogens such as Pythium, Phytophthora, and Rhizoctonia solani requires the introduction into or presence of edaphic sources of organic nutrients in soil for sustenance of biocontrol agents. The decomposition level of organic matter critically affects the composition of bacterial taxa as well as the populations and activities of biocontrol agents. Competition, antibiosis, parasitism, and systemic induced resistance are all affected. Highly stabilized sources of Sphagnum peat consistently fail to support sustained biological control, even when inoculated with biocontrol agents. Composts, on the other hand, can serve as an ideal food base for biocontrol agents and offer an opportunity to introduce and establish specific biocontrol agents into soils, which in turn leads to sustained biological control based on the activities of microbial communities.

Journal Article↗

Antibiotic and biosurfactant properties of cyclic lipopeptides produced by fluorescent Pseudomonas spp. from the sugar beet rhizosphere.

Cyclic lipopeptides (CLPs) with antibiotic and biosurfactant properties are produced by a number of soil bacteria, including fluorescent Pseudomonas spp. To provide new and efficient strains for the biological control of root-pathogenic fungi in agricultural crops, we isolated approximately 600 fluorescent Pseudomonas spp. from two different agricultural soils by using three different growth media. CLP production was observed in a large proportion of the strains (approximately 60%) inhabiting the sandy soil, compared to a low proportion (approximately 6%) in the loamy soil. Chemical structure analysis revealed that all CLPs could be clustered into two major groups, each consisting of four subgroups. The two major groups varied primarily in the number of amino acids in the cyclic peptide moiety, while each of the subgroups could be differentiated by substitutions of specific amino acids in the peptide moiety. Production of specific CLPs could be affiliated with Pseudomonas fluorescens strain groups belonging to biotype I, V, or VI. In vitro analysis using both purified CLPs and whole-cell P. fluorescens preparations demonstrated that all CLPs exhibited strong biosurfactant properties and that some also had antibiotic properties towards root-pathogenic microfungi. The CLP-producing P. fluorescens strains provide a useful resource for selection of biological control agents, whether a single strain or a consortium of strains was used to maximize the synergistic effect of multiple antagonistic traits in the inoculum.

Agriculture↗

Assessment and management of soil microbial community structure for disease suppression.

Identification of the biological properties contributing to the function of suppressive soils is a necessary first step to the management of such systems for use in the control of soilborne diseases. The development and application of molecular methods for the characterization and monitoring of soil microbial properties will enable a more rapid and detailed assessment of the biological nature of soil suppressiveness. Although suppressive soils have provided a wealth of microbial resources that have subsequently been applied for the biological control of soilborne plant pathogens, the full functional capabilities of the phenomena have not been realized in production agricultural ecosystems. Cultural practices, such as the application of soil amendments, have the capacity to enhance disease suppression, though the biological modes of action may vary from that initially resident to the soil. Plants have a distinct impact on characteristics and activity of resident soil microbial communities, and therefore play an important role in determining the development of the disease-suppressive state. Likewise, plant genotype will modulate these same biological communities, and should be considered when developing strategies to exploit the potential of such a natural disease control system. Implementation of consistently effective practices to manage this resource in an economically and environmentally feasible manner will require more detailed investigation of these biologically complex systems and refinement of currently available methodologies.

Agriculture↗

[Survival rate of Leptospira pomona in the soil at a natural leptospirosis focus].

At the area of a natural focus of leptospirosis (caused by L. pomona) in the Mozdok region of the North Occetic ASSR leptospires were detected by the method of dark field microscopy in 22% of intact soil samples. The presence of pathogenic leptospires in this soil was not confirmed by the method of the biological assay on Syrian golden hamsters. In controlled tests lasting 1-277 days, in 30.4% of cases L. pomona retained their viability, pathogenic and antigenic properties for as long as 74 days, while staying in the soil at the focus of infection with humidity being 15.2-31.4% and pH = 6.7-7.2. 11 Leptospira cultures isolated after staying in the soil retained their pathogenic properties and the death of the animals used in the bioassay from the acute form of leptospiral infection.

Animals↗

Influence of disease-suppressive strains of Streptomyces on the native Streptomyces community in soil as determined by the analysis of cellular fatty acids.

Analysis of cellular fatty acid profiles was used to distinguish among introduced pathogen- suppressive strains and indigenous strains of Streptomyces spp. isolated from soil of field plots established to test the efficacy of Streptomyces strains PonSSII and PonR in the biological control of potato scab. Reference libraries of fatty acid profiles were developed for a collection of known pathogenic strains and the introduced suppressive strains. Population densities of pathogen-related, suppressive, and saprophytic Streptomyces strains were determined from the relationship of field isolates to mean library profiles using cluster analysis and the unweighted pair-group method using arithmetic averages. Community diversity was similarly determined. Streptomyces strains PonSSII and PonR were distinguished from each other and from the pathogen group (which clustered together) based on fatty acid profiles. The introduced, suppressive strains successfully colonized the soil and represented 2-19% of the isolates sampled over 2 years. The introduction of the suppressive strains inhibited the population of strains related to the pathogen library at each sample date; the pathogen population was substantially lower in soil from treatments where the suppressive strains were introduced compared with the nonamended control. At harvest, the pathogen-related population was suppressed 85-93 and 36-44% in 1991 and 1992, respectively, in treatments with the suppressive strains compared with the nonamended control. Diversity of the community was not affected by the introduced strains, and diversity and equitability indices were similar among treatments at any sample time. The inhibition of the pathogen-related population was correlated with a reduction of scab symptoms observed in the field plots into which the suppressive strains were introduced. Implications of a fundamental shift in the pathogen-related population in response to the introduction of the suppressive strains for long-term biological control of potato scab are encouraging.

Fatty Acids↗

The potential role of arbuscular mycorrhizal (AM) fungi in the bioprotection of plants against soil-borne pathogens in organic and/or other sustainable farming systems.

Sustainable farming systems strive to minimise the use of synthetic pesticides and to optimise the use of alternative management strategies to control soil-borne pathogens. Arbuscular mycorrhizal (AM) fungi are ubiquitous in nature and constitute an integral component of terrestrial ecosystems, forming symbiotic associations with plant root systems of over 80% of all terrestrial plant species, including many agronomically important species. AM fungi are particularly important in organic and/or sustainable farming systems that rely on biological processes rather than agrochemicals to control plant diseases. Of particular importance is the bioprotection conferred to plants against many soil-borne pathogens such as species of Aphanomyces, Cylindrocladium, Fusarium, Macrophomina, Phytophthora, Pythium, Rhizoctonia, Sclerotinium, Verticillium and Thielaviopsis and various nematodes by AM fungal colonisation of the plant root. However, the exact mechanisms by which AM fungal colonisation confers the protective effect are not completely understood, but a greater understanding of these beneficial interactions is necessary for the exploitation of AM fungi within organic and/or sustainable farming systems. In this review, we aim to discuss the potential mechanisms by which AM fungi may contribute to bioprotection against plant soil-borne pathogens. Bioprotection within AM fungal-colonised plants is the outcome of complex interactions between plants, pathogens and AM fungi. The use of molecular tools in the study of these multifaceted interactions may aid the optimisation of the bioprotective responses and their utility within sustainable farming systems.

Agriculture↗

[Evaluation of Trichoderma spp. as antagonist of Rhizoctonia solani in vitro and as biocontrol of greenhouse tomato plants].

Five Trichoderma isolates were compared in their ability for controlling Rhizoctonia solani attack to tomato plants in greenhouse and as antagonists of this pathogen in three independent laboratory assays. Four out of five isolates showed biocontrol ability and decreased pathogen growth and survival of its sclerotia in soil. Results suggest that dual cultures in Petri dishes and mycoparasitism assays against R. solani sclerotia may be useful for detecting isolates effective as biological control agents against this pathogen in tomato plants.

Agriculture↗