Streptomycete Populations Research Articles

Isolation and Identification of Streptomyces spp. from Desert and Savanna Soils in Sudan.

The purpose of this study was to investigate streptomycete populations in desert and savanna ecozones in Sudan and to identify species based on 16S rRNA gene sequences. A total of 49 different Streptomyces phenotypes (22 from sites representing the desert and semi-desert ecozone; 27 representing the savanna ecozone) have been included in the study. The isolates were characterized phenotypically and confirmed using 16S rRNA gene sequence analysis. The two ecozones showed both similarities and uniqueness in the types of isolates. The shared species were in cluster 1 (Streptomyces (S.) werraensis), cluster 2 (Streptomyces sp.), cluster 3 (S. griseomycini-like), and cluster 7 (S. rochei). The desert ecozone revealed unique species in cluster 9 (Streptomyces sp.) and cluster 10 (S. griseomycini). Whereas, the savanna ecozone revealed unique species in cluster 4 (Streptomyces sp.), cluster 5 (S. albogriseolus/ S. griseoincarnatus), cluster 6 (S. djakartensis), and cluster 8 (Streptomyces sp.). Streptomycetes are widely distributed in both desert and the savanna ecozones and many of these require full descriptions. Extending knowledge on Streptomyces communities and their dynamics in different ecological zones and their potential antibiotic production is needed.

Stability and microbial community analysis during rotary drum composting of vegetable waste

Disposal of vegetable waste in landfills and illegal sites leads to emission of greenhouse gases and leachate production, thereby imposing major environmental issues. As an alternative, this waste can be successfully recycled for its high nutrient content using composting process. Five trials were made with the ratio of 6:3:1 of trial 1 (50 kg), trial 2 (70 kg), trial 3 (90 kg), trial 4 (120 kg) and trial 5 (150 kg) by adding 10 kg of dry leaves in each of the trials as bulking agent. Due to active microbial population and high biodegradable organic matter in vegetable waste, early thermophilic phase was observed within 18–24 h of the composting process in all of the trials with a maximum of 61.4 °C in trial 3. Total mesophilic heterotrophs were observed in the range of 7.1 × 1011 CFU g−1 and gradually reduced to 2.65 × 106 CFU g−1 at the end of 20 days, which was considered due to prolonged thermophilic phase maintained in trial 3. An average of 54–56 °C temperature was maintained for 7 days in trial 3, with spore-forming population in the order of 3.82 × 109 CFU g−1 contributing to higher organic destruction. The populations of fungus, actinomycetes and streptomycetes were observed to reduce during thermophilic phase and remained in the order of 2.85 × 104, 3.8 × 106 and 4.1 × 105 CFU g−1, respectively, at the end of 20 days. CO2 evolution and OUR were in the order of 0.89 and 0.32 mg g−1 VS d−1, respectively, in trial 3 denoting maximum degradation of organic matter and stabilization of compost. Indicator organisms were found well with the standard limits due to elevated temperature. Combinations of waste materials played a major role in favoring microbial succession. Temperature in the compost system had major effect on the survival of the microbial populations. Elevated temperatures favored higher degradation of organic matter, thereby stabilizing the compost within proposed time of composting and also destructing the indicator pathogens.

Sympatric inhibition and niche differentiation suggest alternative coevolutionary trajectories among Streptomycetes

Soil bacteria produce a diverse array of antibiotics, yet our understanding of the specific roles of antibiotics in the ecological and evolutionary dynamics of microbial interactions in natural habitats remains limited. Here, we show a significant role for antibiotics in mediating antagonistic interactions and nutrient competition among locally coexisting Streptomycete populations from soil. We found that antibiotic inhibition is significantly more intense among sympatric than allopatric Streptomycete populations, indicating local selection for inhibitory phenotypes. For sympatric but not allopatric populations, antibiotic inhibition is significantly positively correlated with niche overlap, indicating that inhibition is targeted toward bacteria that pose the greatest competitive threat. Our results support the hypothesis that antibiotics serve as weapons in mediating local microbial interactions in soil and suggest that coevolutionary niche displacement may reduce the likelihood of an antibiotic arms race. Further insight into the diverse roles of antibiotics in microbial ecology and evolution has significant implications for understanding the persistence of antibiotic inhibitory and resistance phenotypes in environmental microbes, optimizing antibiotic drug discovery and developing strategies for managing microbial coevolutionary dynamics to enhance inhibitory phenotypes.

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.

Culture-Based Assessment of Microbial Communities in Soil Suppressive to Potato Common Scab.

A field in East Lansing, MI, showed a decline of potato common scab compared with an adjacent potato field. To confirm that the decline was due to biological factors, the soil was assayed. In the greenhouse, putative common-scab-suppressive soil (SS) was either treated with various temperatures or mixed with autoclaved SS at various ratios. Pathogenic Streptomyces scabies was incorporated into the treated soil at 106 CFU/cm3 of soil, followed by planting of either potato or radish. Disease severity was negatively correlated with the percentage of SS in the mixture and positively correlated with temperature above 60°C. The soil was screened for four groups of potential antagonists (general bacteria, streptomycetes, fluorescent pseudomonads, and bacilli) pairing in culture with S. scabies. The frequency of antagonistic bacteria in SS was higher than common-scab-conducive soil (CS) in all four groups but only pseudomonads and streptomycetes were significantly higher. The population of pathogenic Streptomyces spp. in the rhizosphere of CS was significantly higher than SS. Dilution plating of CS and SS samples showed no clear trends or differences in populations of total fungi, total bacteria, streptomycetes, fluorescent pseudomonads, and bacilli but terminal restriction fragment polymorphism analysis revealed two distinct microbial communities were present in SS and CS.

Diversity Analysis of Streptomycetes and Associated Phosphotranspherase Genes in Soil

An attempt was made to verify the observation that Streptomyces griseus was prevalent in soil based on isolation work. A genus-specific PCR was developed for Streptomyces based on the housekeeping gene atpD and used to investigate species diversity within selected soils. The presence of S. griseus was investigated to determine coexistence of resistance-only streptomycin phosphotransferase (strA) in the same soil as streptomycin producers. Two additional PCR-based assays were developed; one specific for strA in association with production, the other for more diverse strA and other related phosphotranferases. Both the S. griseus atpD and strA genes were below the PCR detection limit in all soils examined. A number of more diverse phosphotransferase genes were amplified, a minority of which may be associated with streptomycin production. We conclude that neither streptomycin producers nor S. griseus are prevalent in the fresh or chitin and starch-amended soils examined (less than 0.1% of soil actinobacteria). One of the soil sites had received plantomycin (active ingredient: streptomycin) and diversity studies suggested that this altered the streptomycete populations present in the soil.

Comparison of Streptomyces diversity between agricultural and non-agricultural soils by using various culture media.

Streptomycetes play a key role in the sustainability of agriculture and indicates the level of health of soil, especially when considering the richness of them that are involved in biological control of soil borne diseases. 20 different soil samples were taken from agricultural (7) and non-agricultural places (13) and populations of streptomycetes were quantified in order to select the general culture media that had better reflect the changes of these bacteria. The most efficient medium for the isolation of Streptomyces was starch casein agar by the addition of nystatin. Pretreatment of soil samples with CaCO3 (1%) increased the streptomycetes occurring on the isolation plates. To establish a correlation with soil physico-chemical parameters, such as pH, salt, N, P, K, Na, Fe, Zn and Cu were also determined, most of the correlations being significantly positive on the quantification of Streptomyces diversity. Streptomycete counts ranged from a high of 6.7 x 10 6 to a low of 2.3 x 10 6 cfu/g dry soil of non-agricultural soils. Streptomycetes constituted 4.8 to 45.8% of the total culturable bacterial community. Higher streptomycete densities were greatest in non-agricultural soils with an average of 14.0% compared to agricultural soils with average of 10.1%. These results suggest that these bacteria may be represent an unexplored resource for pharmaceutical drug discovery but also may provide additional disease control in agriculture.

Molecular Phylogenetic Classification of Streptomycetes Isolated from the Rhizosphere of Tropical Legume (Paraserianthes falcataria) (L.) Nielsen

Intrageneric diversity of 556 streptomycetes isolated from the rhizosphere of tropical legume was determined by using molecular taxonomic method based on 16S rDNA. A total of 46 isolates were taken to represent 37 colour groups of the isolates. 16S rDNA were amplified and subsequently sequenced and the sequences data were aligned with streptomycete sequences retrieved from the ribosomal data base project (RDP) data. Phylogenetic trees were generated by using the PHYLIP software package and the matrix of nucleotide similarity and nucleotide difference were generated by using PHYDIT software. The results confirmed and extended the value of 16S rDNA sequencing in streptomycete systematic. The 16S rDNA sequence data showed that most of the tested colour group representatives formed new centers of taxonomic variation within the genus Streptomyces. The generic assignment of these organisms was underpinned by 16S rDNA sequence data which also suggested that most of the strains represented new centers of taxonomic variation. The taxonomic data indicate that diverse populations of streptomycetes are associated with the roots of tropical legume (P. falcataria). Therefore, the combination of selective isolation and molecular taxonomic procedures used in this study provide a powerful way of uncovering new centers of taxonomic variation within the genus Streptomyces. Key words: molecular phylogenetic, classification, streptomycetes, rhizosphere, Paraserianthes falcatharia

Management of soil microbial communities to enhance populations of Fusarium graminearum-antagonists in soil

Fusarium head blight (FHB), incited by Fusarium graminearum Schwabe is one of the most devastating diseases of wheat. Primary inoculum generated on crop residue is the driving force of FHB epidemics. Fusarium survival on crop residues is affected by soil microbial antagonists. The incorporation of green manures has been shown to increase the density and diversity of microbes in soils, particularly the density and the pathogen-inhibitory activity of specific bacteria and fungi. Evidence of increased streptomycete populations in soil as a response to green manure incorporation, and their negative effect on the survival of Fusarium oxysporum Schlechtendahl in soil, suggests their potential use to reduce the survival of related pathogens. There is, however, no precedent for the use of green manures to promote indigenous streptomycete populations to control FHB. This study investigated the use of green manures (sorghum–sudangrass hybrid [Sorghum bicolor (L.) Moench–S. bicolor (L.) Moench var. sudanense (Piper)] and common buckwheat [Fagopyrum esculentum (Moench)]) for reducing F. graminearum survival in association with wheat residues. Soil bacterial density, streptomycete density and the density and inhibitory activity of F. graminearum-antagonists were monitored from planting until 3 and 6 months following the incorporation of green manures in greenhouse and field experiments, respectively. The decomposition of wheat residues and survival of Fusarium in residues was also assessed. The use of green manures did not statistically impact the survival of F. graminearum in wheat residue. However, green manures promoted the development of higher densities and antagonistic abilities of F. graminearum-antagonists in soils. Additionally, streptomycete densities and F. graminearum-antagonist densities were significantly and positively correlated with reduced survival of Fusarium. The results of our study suggest that the use of green manures can enhance populations of indigenous soil microorganisms antagonistic to the survival of F. graminearum in wheat residue.

Resident bacteria, nitric oxide emission and particle size modulate the effect of Brassica napus seed meal on disease incited by Rhizoctonia solani and Pythium spp

Amendment of orchard soil with low-glucosinolate Brassica napus (rape) seed meal (RSM) suppresses infection of apple roots by Rhizoctonia solani but increases incidence of Pythium spp. infection. Following incorporation of Brassica sp. seed meals, soils were monitored for changes in populations of selected saprophytic and plant pathogenic microorganisms. When conducted in pasteurized soil, which possessed high numbers of Bacillus spp. and lower than detectable numbers of Streptomyces spp., RSM amendment did not provide control of R. solani. Populations of streptomycetes in RSM-amended soil increased to stable levels >20-fold higher than in non-amended soil. Disease suppressiveness was restored to pasteurized RSM-amended soil by adding any of several Streptomyces strains. Maximal rates of nitrification in orchard soil, determined by nitric oxide emission, were observed within two weeks following RSM amendment and inhibition of nitrification via application of nitrapyrin abolished the capacity of RSM to suppress R. solani infection of apple roots when seedlings were planted one day after soil amendment. Apple seedling mortality and Pythium spp. root infection were highest for seedlings planted immediately following incorporation of B. napus cv. Athena RSM, particularly when meal was added in a flake rather than powder form. Lower infection frequencies were observed for seedlings planted four weeks after RSM incorporation, even for soil in which densities of culturable Pythium spp. had not declined. Our results demonstrate that suppression of Rhizoctonia root rot in response to RSM amendment requires the activity of the resident soil microbiota and that initial disease control is associated with the generation of nitric oxide through the process of nitrification.

Distribution of streptomycin resistance and biosynthesis genes in streptomycetes recovered from different soil sites

Streptomycin resistant streptomycetes were isolated from four diverse sites. Two sites in Germany were sampled and included an agricultural experimental plot of apple trees which had a history of streptomycin application (AR) and a control site without antibiotic application (CR). Two sites in the UK were sampled; a pastureland site which had sewage injection (DW) and an agricultural site (CW). The actinomycete counts indicated a high proportion of streptomycin resistant isolates in the CW soil. Streptomycetes were identified by partial sequencing of the 16S rDNA. PCR product of the hypervariable gamma region of 16S rDNA allowed analysis by denaturing gradient gel electrophoresis to assess the diversity within the isolates. The streptomycin and sewage sludge treated sites showed decreased diversity within streptomycete populations. Isolates were screened for the streptomycin resistance gene, strA, and flanking biosynthesis gene, strB1. Distribution of these genes indicated the prevalence of str genes in the streptomycin treated soil. Evidence of horizontal gene transfer was recorded in isolates identified as Streptomyces platensis recovered from CR and AR sites which had acquired a streptomycin resistance gene homologous to that found in Streptomyces griseus. Members of the latter species were the most abundant streptomycin resistant streptomycetes isolated from all soils.

Distribution of streptomycin resistance and biosynthesis genes in streptomycetes recovered from different soil sites

Streptomycin resistant streptomycetes were isolated from four diverse sites. Two sites in Germany were sampled and included an agricultural experimental plot of apple trees which had a history of streptomycin application (AR) and a control site without antibiotic application (CR). Two sites in the UK were sampled; a pastureland site which had sewage injection (DW) and an agricultural site (CW). The actinomycete counts indicated a high proportion of streptomycin resistant isolates in the CW soil. Streptomycetes were identified by partial sequencing of the 16S rDNA. PCR product of the hypervariable γ region of 16S rDNA allowed analysis by denaturing gradient gel electrophoresis to assess the diversity within the isolates. The streptomycin and sewage sludge treated sites showed decreased diversity within streptomycete populations. Isolates were screened for the streptomycin resistance gene, strA, and flanking biosynthesis gene, strB1. Distribution of these genes indicated the prevalence of str genes in the streptomycin treated soil. Evidence of horizontal gene transfer was recorded in isolates identified as Streptomyces platensis recovered from CR and AR sites which had acquired a streptomycin resistance gene homologous to that found in Streptomyces griseus. Members of the latter species were the most abundant streptomycin resistant streptomycetes isolated from all soils.

Occurrence and diversity of plasmids in populations of streptomycetes in soil.

Studies were made of naturally occurring plasmids hosted in Streptomyces strains isolated from two different terrestrial ecosystems: an agricultural field and a protected forest area. Six out of the 147 screened isolates contained plasmids. The strains containing these plasmids were all isolated from the agricultural soil. Plasmids were not found among the strains isolated from the forest area. Cross hybridization of the six newly isolated plasmids revealed very high similarities between four of them. However, no similarities were found between the six newly isolated plasmids and well studied streptomycete plasmids such as pIJ101 and SCP2*. The host strains of the four similar plasmids belonged to three different species S. anulatus, S. rochei and S. diastaticus. This implies a possible conjugative transfer of these plasmids within the streptomycete population in the agricultural area. The reason for the absence of streptomycete plasmids from the populations derived from the forest area is discussed.

Evidence for transfer of antibiotic-resistance genes in soil populations of streptomycetes.

Phylogenetic analysis was used to evaluate the hypothesis of gene transfer in streptomycetes, many of which are antibiotic producers. The diversity and possible origins of streptomycin-resistance genes was investigated for a population of Streptomyces strains isolated from a site in Brazil where antibiotic production had previously been implicated The analysis provides compelling evidence for the transfer of these genes. Examination of other Streptomyces-type strains also reveals a scattered distribution of streptomycin producers with respect to the overall phylogeny. These results suggest that horizontal gene transfer may be an important factor in the evolution of antibiotic genes in streptomycetes.

Interrelations of micro-organisms and mulberry

The mulberry leaves were shown to harbour substantial populations of bacteria, streptomycetes, yeasts, and moulds. Azotobacter and Beijerinckia were observed to contribute to nearly 5 to 10 per cent of the bacterial population. When grown in water culture under sterile conditions, Azotobacter inoculation on the leaf or root surface was found to increase plant growth, dry wt, and nitrogen content of the mulberry. The beneficial effect of Azotobacter was largely influenced by the presence of a carbon source in the plant nutrient solution. The root inoculation in comparison to leaf application was found to confer greater benefits to the growing plant. The presence of carbohydrates and amino acids in the leaf leachates of mulberry was shown. The mutual beneficial nature of the association of the plant and Azotobacter has been brought to light.

Effects of Prescribed Burning on Soil Microorganisms in a Minnesota Jack Pine Forest

Soil dilution plate counts on selective media for bacteria, streptomycetes, and fungi were made at intervals before and after two prescribed burns on 10—acre jack pine tracts. Carbon dioxide determinations were also made. These tracts were compared with similar tracts cut and unburned and uncut and unburned. Three—year results are given. The number and activity of most microorganisms decreased immediately after fire but rose abruptly to a very high level after the first rainfall following burning. This increase is believed caused by the leaching of ash minerals. Depth and extent of the effects were influenced by intensity of fire and moisture conditions. Number and activity of organisms were generally lower in burned soil the second growing season after fire. Some effects were still noticeable the third growing season. Streptomycete populations were greatly increased the third growing season. Rainfall also caused a fluctuation of microbial populations on the unburned tracts.