Soil Streptomycetes Research Articles

Development Strategy for the New Environmentally Friendly Multifunctional Bioformulations Based on Soil Streptomycetes

Development Strategy for the New Environmentally Friendly Multifunctional Bioformulations Based on Soil Streptomycetes

Impact of metabolic biopreparations based on soil streptomycetes on productivity of spring wheat

It has been established the new multifunctional metabolic bioformulations based on a set of biologically active substances from soil sreptomycetes Avercom, with the addition of elicitors of salicylic acid or chitosan, Violar and Phytovit promote the growth of microorganisms of major ecological and functional groups in the root area of spring wheat, and they increase the activity of the enzyme phenylanine- ammonia-lyase (PAL) in plant cells that indicates the induction of plant resistance to pathogens. This reduces the destruction of plants of spring wheat by the root rot and plant-parasitic nematodes. As a consequence of the action of Averсom nova-2, Violar and Phytovit the grain yield increased by 16.9–32.5% relative to control and 10.9–25.7% compared with chemical disinfectant Vitavaks 200 FF. At the same time the quality grain and proficiency were increased. The reduction of pesticide load on agrocenoses, an improvement of the ecological condition and the raise of productivity of agroecosystems can be achieved by using the new multifunctional metabolic bioformulations.

Антинематодна активність метаболітів, що продукуються грунтовими стрептоміцетами

To study in vitro the antinematicide activity of soil streptomycetes metabolite and bioformulations based on them against root-knot, cyst and leaf nematodes of species. Action of streptomycetes metabolites and their bioformulations were determined in vitro against larvae at two age of root-knot nematode M incognita (Kofoid & White, 1919) Chitwood 1949 and sugar beet cyst nematode H. schachtii A. Schmidt 1871, and leaf nematodes ofAphlenchoides genus by cultivation them in solutions of biomass ethanol extracts separate individual components metabolites and bioformulations during 24 hours. Nematicide and nematistatic effects of the substances were evaluated by the change in the motility activity of the nematodes. Soil streptomycetes S. violaceus IMV Ac-5027, S. averinitilis IMV Ac-5015 and metabolite bioformulations based on them (Violar, Aver- com and Avercom-nova, respectively) showed significant nematicidal activity against plant pathogenic nematodes M incognita and H. schachtii. The biomass extracts of S. avermiti- lis IMV Ac-5015 and S. violaceus IMV Ac-5027 caused 100 % death of nematodes, and S. netropsis IMV Ac-5025 caused only 75 % mortality during 24 hours of action. Using an antibiotic complex from S. violaceus IMV Ac-5027 was divided on six fractions that were different from the avermectin complex. Purified fractions of individual antibiotic complex of S. violaceus IMV Ac-5 027 have a negative impact against the root-knot nematode M incognita. The highest nematicidal activity showed fractions #4 and #5 since their efficiency was over 90 % after a 0.5 hours of action. Nematicidal efficiency of fractions ## 1, 2, 3 and 6 remained between 72.6-86.2 % during 24 hours of action. Fractions 4 and 5 showed high nematicidal effectiveness against leaf nematodes of Aphelenchoides genus where 90-95 % level of helminthes death was observed after 4 hours of action. Among created complex metabolite bioformulations Avercom-nova was the most effective which caused 100 % mortality of nematodes M incognita and H. schachtii after 24 hours of action. Bioformu- lation Violar caused 96.6-96.8 and 95.2-97.0 % of death of nematodes M incognita and H. schachtii, after 24 hours of action. Phytovit based on S. netropsis IMV Ac-5025 showed low nematicide activity against of nematodes, which did not exceed 15.2-18.4 % of their death after 24 hour of action. Bioformulations based on streptomycetes metabolites did not inferior to chemical insecticide Konfidor Maxi, which caused the death of 93.3-94.6 % nematodes at doses recommended by the manufacturer. Soil streptomycetes S. violaceus IV Ac-5027 and S. avermitilis IMV Ac-5015 are promising producers for creation of bioformulations with nematicidal activity against plant-parasitic nematode of M incoenita and H. schachtii.

Идентификация и антагонистические свойства почвенного стрептомицета Streptomyces sp. 100

Determination of the taxonomic status of the soil streptomycete Streptomyces sp. 100 and study of its antagonistic properties against phytopathogenic and opportunistic human microorganisms. For the identification of the strain a set of conventional methods morphological and cultural, physiological and biochemical characteristics of the producer, as well as molecular genetic analysis of 16S rRNA gene were used. Streptomycete was cultivated on agar nutrient and liquid soy medium until the stationary phase of growth. The antagonistic activity of the strain was studied by agar diffusion method. The study of morphological and cultural properties showed that Streptomyces sp. 100 formed the colonies with irregular edges protruding from the depressed center, straight sporophores were short, gathered in whorls; spores were oval, smooth shell dispute. Growing on agar medium (pH 6.8-7.4, temperature 28 °C, microaerophilic conditions) this strain formed mycelium of various colors: the air white, white-yellow, white-brown or substrate tan, cream and yellow, creamy carmine, yellow-brown. A soluble pigment was yellow and yellow-brown, while melanoid pigment was not detected. The morphological, cultural, physiological, biochemical and molecular genetic characteristics of the soil streptomycete let to identify it as Streptomyces netropsis (Finlay et al., 1951) IMV Ac-5025 (UCM Ac-2186) that is an active antagonist IMV Ac-5025 against plant pathogens. Growing on a surface of agaric nutrient media it inhibits phytopathogenic bacteria (Xanthomonas axonopodis pv. glycines 8609, Pseudomonas savastanoi pv. glycinea 8571, P. syringae pv. coronafaciens 9030) and fungi (Alternaria alternata 16814, Fusarium оxysporum 54201) zone of growth inhibition were 20 - 32 mm and 16 - 30 mm respectively. The supernatant of culture medium and the ethanol extract of biomass inhibited the growth of pathogenic bacteria and fungi. The most sensitive to action of a supernatant of cultural liquid were P. syringae pv. atrofaciens 7886 and Clavibacter michiganensis ssp. michiganensis 102, growth inhibition zones - 42 and 30 mm respectively. It should be noted that in the majority of cases the supernatant of cultural liquid suppressed growth of phytopathogenic bacteria in comparison with biomass extract more actively. At the same time only biomass extract inhibited the growth of P. syringae pv. coronafaciens 9060, P. corrugatа 9070, X. anoxopodis pv. glycines 9075, X. anoxopodis pv. glycines 8609 and Pantoea agglomerans 8490. Tolerant to metabolites of S. netropsis IMV Ac-5025 were P. syringae pv. atrofaciens 8291 and X. visicatoriae 7790. The extract of biomass S. netropsis IMV Ac-5025 inhibited growth of all studied strains of phytopathogenic fungi (A. alternata 16814, A. culmorum 00790, F. оxysporum 54201, F. tricinetum 00795, F. oxysporum n.33, Cladosporium herbarum 16863, Cochliobolus spicifas 16860, Nigrospora oryzae 16864). The supernatant of the cultural liquid also showed the oppressing action on fungi, except for Cladosporium herbarum 16863 and Cochliobolus spicifas 16860. The strain was almost ineffective against opportunistic human microorganisms (Escherichia coli ATCC 25922, Bacillus pumilus NCTC 8241, Staphylococcus aureus FDA 209P et al.). The lack of action of Streptomyces netropsis IMV Ac-5025 on the opportunistic human microorganisms and the active antagonism of phytopathogens, both, define potential its application for plant protection.

CHARACTERISTIC OF SELECTED SOIL STREPTOMYCETES WITH ANTIMICROBIAL POTENTIAL AGAINST PHYTOPATHOGENIC MICROORGANISMS

The main objective of the present study was isolation and identification of soil streptomycetes, having antibacterial and antifungal activity against 8 selected phytopathogenic microorganisms, which attack crops in Slovakia. Out of 27 morphologically different streptomycete isolates, 14 of them demonstrated antimicrobial activity against at least two tested microorganisms in primary screening. Ethylacetate extract used for secondary screening showed different inhibitory pattern in comparison with primary screening. The tested isolates were mostly active against Gram-positive bacterium Clavibacter michiganensis subps. sepedonicus, the causal agent of bacterial canker of tomaten and against fungus Fusarium poae, pathogen of cereals, which can also infest stored grains. Only three isolates, namely VY59, VY87 and VY47 showed broad spectrum of inhibition activity and therefore were used for further identification. On the basis of various morphological (color of aerial and substrate mycelium, growth, production of pigments and melanin on ISP media), physiological (optimal pH, temperature, NaCl tolerance and C-utilization) and biochemical (ApiZym and ApiCoryne stripes) methods these strains belonged to the genus Streptomyces. In addition to the antimicrobial profile, the strains differed in API ZYM test results, which imply that the selected strains might produce different antimicrobial substances. Throught the comparative analysis of 16S rRNA gene, the most active isolates contained different nucleotide sequences for the 16S rRNA gene. Sequence similarity search by BLAST program revealed that they show sequence similarities to Streptomyces somaliensis (VY59), Streptomyces sp. (VY87) and Streptomyces albidoflavus (VY47). These three isolates with broader spectrum of antimicrobial activity can be used in the development of substances for agriculture purposes.

Extracellular synthesis gold nanotriangles using biomass of Streptomyces microflavus.

Applications of nanotechnology and nano-science have ever-expanding breakthroughs in medicine, agriculture and industries in recent years; therefore, synthesis of metals nanoparticle (NP) has special significance. Synthesis of NPs by chemical methods are long, costly and hazardous for environment so biosynthesis has been developing interest for researchers. In this regard, the extracellular biosynthesis of gold nanotriangles (AuNTs) performed by use of the soil Streptomycetes. Streptomycetes isolated from rice fields of Guilan Province, Iran, showed biosynthetic activity for producing AuNTs via in vitro experiments. Among all 15 Streptomyces spp. isolates, isolate No. 5 showed high biosynthesis activity. To determine the bacterium taxonomical identity at genus level, its colonies characterised morphologically by use of scanning electron microscope. The polymerase chain reaction (PCR) molecular analysis of active isolate represented its identity partially. In this regard, 16S rRNA gene of the isolate was amplified using universal bacterial primers FD1 and RP2. The PCR products were purified and sequenced. Sequence analysis of 16S rDNA was then conducted using National Center for Biotechnology Information Basic Local Alignment Search Tool method. The AuNTs obtained were characterised by ultraviolet-visible spectroscopy, atomic force microscopy, transmission electron microscopy and energy dispersive X-ray spectroscopy, Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction spectroscopy analyses. The authors results indicated that Streptomyces microflavus isolate 5 bio-synthesises extracellular AuNTs in the range of 10-100 nm. Synthesised SNPs size ranged from 10 to 100 nm. In comparison with chemical methods for synthesis of metal NPs, the biosynthesis of AuNTs by Streptomyces source is a fast, simple and eco-friendly method. The isolate is a good candidate for further investigations to optimise its production efficacy for further industrial goals in biosynthesis of AuNTs.

Effect of fertilization on biological activity of community of soil streptomycetes

The search for new natural mechanisms to inhibit the growth of phytopathogenic microorganisms has become widely widespread. Therefore, the main objective of the present study was determination of antimicrobial activities of actinomycetes isolated from agricultural soil, which was fertilized mainly by organic fertilizers, against 8 selected phytopathogenic strains. Among the actinomycetes, Streptomyces species have been extensively studied, because they have been recognized as an important source of secondary metabolites, which can suppress the growth of undesirable pests in crops. The results indicated that the richest source of Streptomyces colonies was soil fertilized with compost (103 x104 CFU*g-1 dry soil). On the basis of morphological signs, total of 65 isolates were selected and examined for antimicrobial activities. Isolates exhibited the best activity against Gram negative bacterium Clavibacter michiganensis subsp. sepedonicus, disease agent of “ring rot” of potatoes and against fungus Fusarium poae, disease agent of Fusarium head blight of wheat. Twelve isolates exhibited promising broad-spectrum activity against tested organisms. On the basis of results, six of them were selected for further screening. Comparison of polyphasic studies with available literature led to identification of biological active strains S. olivochromogenes (13SC11), S. avermitilis (13SC2), S. rishiriensis (13SC13), S. globisporus (13SC19), S. sampsonii (13SPC10) and S. avidinii (13SPC4). After quantification analysis of various enzymes, tested isolates produced alkaline phosphatase, leucinearylamidase, valinearylamidase, acid phosphatase, naphtol-AS-BI-phosphohydrolase, glucosidase in high values (>40 nmol) and were positive for nitrate reduction, hydrolysis of gelatin, urease, and esculin. These isolates can be used in the development of new biopesticides anf biofertilizers with antibacterial and antifungal effect.

AUXINS BIOSYNTHESIS BY SOIL STREPTOMYCES – THE ANTAGONISTS OF PHYTOPATHOGENIC MICROORGANISMS AND NEMATODES

Aim. Research of the auxins biosynthesis by soil streptomycetes – the antagonists of phytopathogenic microorganismses and plant parasitic nematodes on synthetic and organic nutrient mediums. Methods. Auxins were determined by the quantitative of spectrodensitometric thin layer chromatography in the supernatants of cultural liquid and ethanol extracts of biomass. Results. In the stationary growth phase in the supernatants of cultural liquid and biomass of S. netropsis UCM Ac-2186 and S. violaceus UKM Ac-2191 accumulated a significant amount of auxins. Besides the basic active forms of indole-3-acetic acid and indole-3-butyric acid the metabolites contained the indole-3-acetic acid hydrazide, indole-3-carbinol, indole-3-carboxaldehyde and indole-3-carboxylic acid. Conclusions. The plant growth regulate activity of metabolite bioformulations developed on the basis of soil streptomycetes is largely due to their increased ability to auxins biosynthesis, t particularly indole-3-acetic acid and indole-3-butyric acid.

Molecular and functional characteristics of streptomycete communities in relation to soil factors and potato common scab

Naturally-occurring disease-suppressive soils provide control of plant pathogens via the activities of indigenous microbes. While all soils contain antagonistic microbes, there is little systematic understanding of the correlates of variation in indigenous antagonist populations and on their relationships with plant diseases. We characterized the population densities, inhibitory capacities, and phylogenetic composition of soil streptomycete communities in a potato field over time in relation to edaphic factors, antagonist inoculation, and potato common scab severities. Antagonistic Streptomyces populations were highly variable in time and space. Similarly, metagenomic analyses of streptomycete communities showed extensive spatial and temporal variation in composition. Soil characteristics (pH, potassium, organic matter, nitrate, and phosphorous) sometimes explained up to 50% of the spatial variation in Streptomyces population densities, proportion of inhibitory isolates, or pathogen suppressive capacity among locations in the field. Soil pH was positively correlated with common scab severity, and negatively correlated with the proportion of pathogen-inhibitory Streptomyces among locations in the field. This suggests that high pH may have direct beneficial effects on pathogen populations and disease development, and/or indirect effects on pathogen populations via reductions in pathogen-inhibitory Streptomyces populations. Mean pathogen suppressive capacity of antagonistic Streptomyces was negatively correlated with common scab disease. Streptomyces inoculants had no discernible effect on streptomycete community composition, antagonistic capacities, or potato common scab severities, likely reflecting poor colonization of inoculants. Further understanding of the relationships between indigenous antagonist populations and plant diseases will be important to harnessing the potential of indigenous communities to contribute to sustainable disease management.

Streptomyces somaliensis mediated green synthesis of silver nanoparticles

Objective(s): The development of reliable and ecofriendly process for the synthesis of nano-metals is an important aspect in the field of nanotechnology. Nano-metals are a special group of materials with broad area of applications. Materials and Methods: In this study, extracellular synthesis of silver nanoparticles (SNPs) performed by use of the gram positive soil Streptomycetes. Streptomycetes isolated from rice fields of Guilan Province, Iran (5 isolates). Initial characterization of SNPs was performed by visual change color. To determine the bacterium taxonomical identity, its colonies characterized morphologically by use of scanning electron microscope. The PCR molecular analysis of active isolate represented its identity partially. In this regard, 16S rDNA of isolate G was amplified using universal bacterial primers FD1 and RP2. The PCR products were purified and sequenced. Sequence analysis of 16S rDNA was then conducted using NCBI GenBank database using BLAST. Also SNPs were characterized by, transmission electron microscopy (TEM) and X-ray diffraction spectroscopy (XRD). Results: From all 5 collected Streptomyces somaliensis isolates, isolate G showed highest extracellular synthesis of SNPs via in vitro. SNPs were formed immediately by the addition of (AgNO3) solution (1 mM). UV-visible spectrophotometry for measuring surface plasmon resonance showed a single absorption peak at 450 nm, which confirmed the presence of SNPs. TEM revealed the extracellular formation of spherical silver nanoparticles in the size range of 5-35 nm. Conclusions: The biological approach for the synthesis of metal nanoparticles offers an environmentally benign alternative to the traditional chemical and physical synthesis methods. So, a simple, environmentally friendly and cost-effective method has been developed to synthesize AgNPs using Streptomycetes.

Фiтозахиснi та рiстрегулювальнi властивостi метаболiтних препаратiв на основi грунтових стрептомiцетiв

Фiтозахиснi та рiстрегулювальнi властивостi метаболiтних препаратiв на основi грунтових стрептомiцетiв

Mitrecin A , an endolysin‐like bacteriolytic enzyme from a newly isolated soil streptomycete

An open reading frame with homology to known endolysin genes was identified in the genome of Streptomyces sp. strain 212, which is a newly isolated soil bacterium. The heterologously expressed gene product of this endolysin-like gene, called Mitrecin A, demonstrated bacteriolytic activity against several Gram-negative bacteria. The genome of the bacterial strain was sequenced to draft quality using pyrosequencing followed by genome assembly and gene annotation. Within the sequence, a chromosomally located endolysin-like open reading frame was predicted. The gene product, designated Mitrecin A, was heterologously expressed and isolated from contaminating proteins as a fusion protein to a 6-histidine tag. Mitrecin A consists of 127 amino acids arranged in modular domains of activity. It has an estimated molecular weight of 14·3 kDa and retains sequence homology to the M15C peptidase subfamily of zinc metallocarboxypeptidases. The heat-labile purified recombinant protein has an overall positive charge, has optimal catalytic activities at 26°C in solution of pH 9 with 1% saline and has bacteriolytic activity against Gram-negative bacteria of the medically important genera Aeromonas, Escherichia, Salmonella, Shigella, Vibrio and Yersinia.Significance and Impact of the StudyThe gene of a new protein antimicrobial, Mitrecin A, was discovered in the genome of a soil bacterium. The purified recombinant enzyme, resulting from heterologous over expression of the gene, was found to be tolerant of increased pH conditions and to have bacteriolytic activity against Gram-negative bacteria of the medically important genera Aeromonas, Escherichia, Salmonella, Shigella, Vibrio and Yersinia. Characterization of enzymes such as Mitrecin A from previously uncharacterized bacteria provides potential options for new biocontrol agents in medically and economically important applications like therapeutics, disinfectants, food preservatives, agricultural livestock antimicrobials, and inhibitors of biofilm production.

Growth of streptomycetes in soil and their impact on bioremediation

The impact of the extremely heavy metal resistant actinomycete Streptomyces mirabilis P16B-1 on heavy metal mobilization/stabilization, phytoremediation and stress level of plants was analyzed in the presence and absence of Sorghum bicolor in sterile microcosms containing highly metal contaminated or control soil. For control, a metal sensitive S. lividans TK24 was used. The metal contents with respect to the mobile and specifically adsorbed fractions of the contaminated soil were considerably decreased by addition of both, living and dead biomass of the strains, with the heavy metal resistant S. mirabilis P16B-1 showing considerably higher impact. Both strains could grow in control soil, while only S. mirabilis P16B-1 formed new tip growth in the metal contaminated soil. A plant growth promoting effect was visible for S. mirabilis P16B-1 in contaminated soil enhancing the dry weight of inoculated Sorghum plants. Thus, metal resistant strains like S. mirabilis P16B-1 are able to enhance phytoremediation of heavy metal contaminated soils.

Isolation and characterization of soil Streptomyces species as potential biological control agents against fungal plant pathogens

The use of antagonist microorganisms against fungal plant pathogens is an attractive and ecologically alternative to the use of chemical pesticides. Streptomyces are beneficial soil bacteria and potential candidates for biocontrol agents. This study reports the isolation, characterization and antagonist activity of soil streptomycetes from the Los Petenes Biosphere Reserve, a Natural protected area in Campeche, Mexico. The results showed morphological, physiological and biochemical characterization of six actinomycetes and their inhibitory activity against Curvularia sp., Aspergillus niger, Helminthosporium sp. and Fusarium sp. One isolate, identified as Streptomyces sp. CACIS-1.16CA showed the potential to inhibit additional pathogens as Alternaria sp., Phytophthora capsici, Colletotrichum sp. and Rhizoctonia sp. with percentages ranging from 47 to 90%. This study identified a streptomycete strain with a broad antagonist activity that could be used for biocontrol of plant pathogenic fungi.

Taking nature into lab: biomineralization by heavy metal-resistant streptomycetes in soil

Abstract. Biomineralization by heavy metal-resistant streptomycetes was tested to evaluate the potential influence on metal mobilities in soil. Thus, we designed an experiment adopting conditions from classical laboratory methods to natural conditions prevailing in metal-rich soils with media spiked with heavy metals, soil agar, and nutrient-enriched or unamended soil incubated with the bacteria. As a result, all strains were able to form struvite minerals (MgNH4PO4• 6H2O) on tryptic soy broth (TSB)-media supplemented with AlCl3, MnCl2 and CuSO4, as well as on soil agar. Some strains additionally formed struvite on nutrient-enriched contaminated and control soil, as well as on metal contaminated soil without addition of media components. In contrast, switzerite (Mn3(PO4)2• 7H2O) was exclusively formed on minimal media spiked with MnCl2 by four heavy metal-resistant strains, and on nutrient-enriched control soil by one strain. Hydrated nickel hydrogen phosphate was only crystallized on complex media supplemented with NiSO4 by most strains. Thus, mineralization is a dominant property of streptomycetes, with different processes likely to occur under laboratory conditions and sub-natural to natural conditions. This new understanding might have implications for our understanding of biological metal resistance mechanisms. We assume that biogeochemical cycles, nutrient storage and metal resistance might be affected by formation and re-solubilization of minerals like struvite in soil at microscale.

Biotransformation of the Fungal Phytotoxin Fomannoxin by Soil Streptomycetes

Rhizosphere-associated Streptomyces sp. AcH 505 (AcH 505) promotes infection of Norway spruce (Picea abies) with the pathogenic fungus Heterobasidion abietinum 331, while Streptomyces sp. GB 4-2 (GB 4-2) enhances spruce defense against the fungus. To identify whether these bacteria influence the availability of the fungal phytotoxin fomannoxin and hence spruce infection, we analyzed the fomannoxin yield in H. abietinum 331-AcH 505 dual cultures. Further, the fate of fomannoxin was studied by adding the compound to cultures of AcH 505, GB 4-2 and nine other soil streptomycetes. Culture filtrates were extracted with ethyl acetate and analyzed by HPLC. Structures of novel compounds were elucidated by HPLC-HR-ESI-Orbitrap-MS and NMR spectroscopy. Phytotoxicity of the compounds was determined by in vivo measurement of maximum photosystem II efficiency of Arabidopsis thaliana seedlings. The amount of fomannoxin in H. abietinum 331-AcH 505 dual cultures was reduced compared to axenic fungus cultures and fungus-plant dual cultures. Following addition of fomannoxin to AcH 505 cultures, the compound disappeared and three novel fomannoxin derivatives without phytotoxic activity were detected. Another novel compound, fomannoxin amide, was discovered following fomannoxin addition to GB 4-2 cultures. Nine other streptomycetes converted fomannoxin into fomannoxin acid or fomannoxin amide. Both compounds exhibit the same phytotoxicity as fomannoxin. We, thus, conclude that the streptomycete-mediated modulation of spruce infection with H. abietinum 331 does not depend on the availability of fomannoxin. We further add evidence to the observation that the lipophilic side chain of fomannoxin is an important structural element for its phytotoxicity.

Use of soil streptomycetes from Maseno (Kenya) to inhibit growth of Pyricularia grisea invitro cultures.

The most serious biotic constraint to finger millet worldwide is blast caused by the fungus Pyricularia grisea. Its management has been very challenging. However not a formidable control solution has been offered and therefore a need for other approachs. Actinomycetes belong to the phylum proteobacteria, they are gram +ve, terrestrial or aquatic with high guanine and cytosine content. Soil actinomycetes have antagonistic activity against a wide range of plant pathogens.This study isolated and characterizes some soil actinomycetes in Maseno area (Western Kenya) and studied their antifungal effect on P. grisea pathogenic on finger millet. Soil sample collected was dissolved in distilled water, homogenized and spread over modified Kuster's agar. Colonies of actiomycetes were isolated as from the fourth day of incubation. . Finger millet heads and leaves displaying symptoms of blast were collected and Pyricularia grisea isolated on autoclaved powered finger-millet debris and maintained on PDA. A single steak of actinomycetes culture was made on a fresh Kuster's agar and incubated for 3-4 days at 26 0 C -28°C, and P. grisea, 20 mm, at right angles to the original streak of actinomycetes. The results indicate that isolates AT2 and AT4 are pathogenic to Pyricularia grisea and can be a basis for bio-control of blast in finger millet.

Effects of plant host species and plant community richness on streptomycete community structure

We investigated soil streptomycete communities associated with four host plant species (two warm season C4 grasses: Andropogon gerardii, Schizachyrium scoparium and two legumes: Lespedeza capitata, Lupinus perennis), grown in plant communities varying in species richness. We used actinobacteria-selective PCR coupled with pyrosequencing to characterize streptomycete community composition and structure. The greatest pairwise distances between communities were observed in contrasts between monocultures of different plant species, indicating that plant species exert distinct selective effects on soil streptomycete populations. Increasing plant richness altered the composition and structure of streptomycete communities associated with each host plant species. Significant relationships between plant community characteristics, soil edaphic characteristics, and streptomycete community structure suggest that host plant effects on soil microbial communities may be mediated through changes to the soil environment. Co-occurring streptomycete taxa also shared consistent relationships with soil edaphic properties, providing further indication of the importance of habitat preference for taxon occurrence. Physical distance between sampling points had a significant influence on streptomycete community similarity. This work provides a detailed characterization of soil streptomycete populations across a field scale and in relation to plant host identity and plant community richness.

Imaging secondary metabolism of Streptomyces sp. Mg1 during cellular lysis and colony degradation of competing Bacillus subtilis

Soil streptomycetes are saprotrophic bacteria that secrete numerous secondary metabolites and enzymes for extracellular functions. Many streptomycetes produce antibiotics thought to protect vegetative mycelia from competing organisms. Here we report that an organism isolated from soil, Streptomyces sp. Mg1, actively degrades colonies and causes cellular lysis of Bacillus subtilis when the organisms are cultured together. We predicted that the inhibition and degradation of B. subtilis colonies in this competition depends upon a combination of secreted factors, including small molecule metabolites and enzymes. To begin to unravel this complex competitive phenomenon, we use a MALDI imaging mass spectrometry strategy to map the positions of metabolites secreted by both organisms. In this report, we show that Streptomyces sp. Mg1 produces the macrolide antibiotic chalcomycin A, which contributes to inhibition of B. subtilis growth in combination with other, as yet unidentified factors. We suggest that efforts to understand competitive and cooperative interactions between bacterial species benefit from assays that pair living organisms and probe the complexity of metabolic exchanges between them.

Plant community effects on the diversity and pathogen suppressive activity of soil streptomycetes

Ecological factors that promote pathogen suppressive microbial communities remain poorly understood. However, plants have profound impacts on the structure and functional activities of soil microbial communities, and land-use changes which alter plant diversity or community composition may indirectly affect the structure and function of microbial communities. Previous research has suggested that the streptomycetes are significant contributors to pathogen suppression in soils. We compared soil streptomycete communities from high and low plant diversity treatments using an experimental manipulation that altered plant diversity while controlling for soil structure and disturbance. Specifically, we characterized an isolate collection for inhibition of plant pathogens as a measure of functional activity, and for 16S rDNA sequence to measure community structure. In this system, high and low diversity plant communities supported streptomycete communities with similar diversity, phylogenetic composition, and pathogen suppressive activity. However, inhibitory phenotypes differed among treatments for several phylogenetic groups, indicating that local selection is leading to divergence between streptomycetes from high and low plant diversity communities. Although the ability to inhibit plant pathogens was common among soil streptomycetes, pathogen-inhibitory activity differed widely among phylogenetic groups. The breadth and intensity of pathogen inhibition by soil streptomycetes were positively related.