scholarly journals Seed-Colonizing Bacterial Communities Associated with the Suppression of Pythium Seedling Disease in a Municipal Biosolids Compost

2012 ◽  
Vol 102 (5) ◽  
pp. 478-489 ◽  
Author(s):  
Mei-Hsing Chen ◽  
Allison L. H. Jack ◽  
I. Cristina McGuire ◽  
Eric B. Nelson
Keyword(s):  
Restriction Fragment ◽  
Bacterial Communities ◽  
Rna Extraction ◽  
Pythium Ultimum ◽  
Disease Suppression ◽  
Seedling Disease ◽  
Treated Seed ◽  
Municipal Biosolids ◽  
The Impact ◽  
Biosolids Compost

This study was designed to characterize seed-colonizing microbial communities that were previously shown to be involved in the suppression of seedling disease caused by Pythium ultimum in a municipal biosolids compost. Selective microbial inhibitors were employed to inactivate portions of the microbial community associated with seed germinated in a compost medium to evaluate their impact on disease suppression. After initial screenings for toxicity to both cucumber and P. ultimum, six selective inhibitors were eventually used to assess the impact of seed treatment on the reduction of bacterial and fungal populations and on disease suppression. Rifampicin was the most effective inhibitor for inactivating disease suppression. Bacterial communities that colonized cucumber seed sown in compost medium for 8 h and seed sown in compost medium for 8 h followed by a 3-h treatment of either rifampicin at 500 ppm or water were dislodged from seed surfaces and subjected to RNA extraction and reverse transcription to cDNA. Differences in the composition of seed-colonizing bacterial communities were assessed using terminal restriction fragment length polymorphisms (T-RFLP) of polymerase chain reaction-amplified 16S rDNA genes. T-RFLP profiles revealed a diversity of distinct bacterial taxa, a number of which dominate seed surfaces within 8 h of sowing. Analysis of similarity (ANOSIM) using terminal restriction fragment (T-RF) presence or absence showed that community profiles of nontreated and water-treated seed were quite similar whereas community profiles from rifampicin-treated seed were distinct. Differences in community profiles based on T-RF abundance (peak height and peak area) indicated that all treatments were unique (ANOSIM, all pairwise comparisons P < 0.05) Peaks heights and areas of relatively few T-RFs were reduced to zero following rifampicin treatment and 34 T-RFs explained 85% of the observed difference between treatments. Tentative taxon assignments for each of the T-RFs that contributed to the treatment differences revealed a preponderance of sequences with affinities to the α-, β-, and γ-Proteobacteria and Firmicutes. Limited sequencing of clones associated with water-treated and rifampicin-treated seed revealed the presence of similar taxa dominated by members of the γ-Proteobacteria. Many species within these taxa (such as Pseudomonas spp., Enterobacter spp., and Bacillus spp.) are known to be suppressive to Pythium diseases. Results of our study have confirmed that Pythium disease suppression in a municipal biosolids compost is mediated by compost-associated bacteria that colonize seed within hours after sowing. By focusing on actively growing microbes in the infection court during important stages of pathogen infection, we believe we can more efficiently determine the mechanisms of disease suppression and the microbes involved. Although specific to this pathosystem and compost, our results have a much broader scope of inference and illustrate the utility of such a targeted approach in identifying a relatively small subset of microbial taxa from complex communities likely to be involved in disease suppression.

scholarly journals Seed-Colonizing Microbes from Municipal Biosolids Compost Suppress Pythium ultimum Damping-Off on Different Plant Species

2008 ◽  
Vol 98 (9) ◽  
pp. 1012-1018 ◽  
Author(s):  
M.-H. Chen ◽  
E. B. Nelson
Keyword(s):  
Plant Species ◽  
Pythium Ultimum ◽  
Disease Suppression ◽  
Microbial Consortia ◽  
Damping Off ◽  
Microbial Species ◽  
Pathogen Suppression ◽  
Municipal Biosolids ◽  
Leaf Compost ◽  
Biosolids Compost

Composts are known for their suppressive properties toward many different seed- and root-infecting pathogens and diseases. Although disease and pathogen suppression induced by composts is believed to be mediated by microbial activities, the nature of the microbial species and processes responsible for suppressiveness remain unknown. We demonstrated previously that seed-colonizing microbial consortia from leaf compost could explain the observed levels of Pythium ultimum-induced damping-off suppression on cotton. The aim of the present work was to determine whether seed-colonizing microbial consortia could explain Pythium damping-off suppression in municipal biosolids compost on three different plant species. Significant levels of disease suppression were observed on cucumber, wheat, and pea at water potentials of –2 kPa. The suppression of damping-off on cucumber and wheat could be eliminated by autoclaving the compost prior to sowing. High levels of suppressiveness were expressed both on cucumber and on wheat seed surfaces within 8 h of sowing. However, the expression of damping-off suppression on the surface of pea seeds was inconsistent and highly variable. Our results demonstrate that compost-induced suppression of P. ultimum damping-off of cucumber and wheat can be explained by the microbial consortia colonizing seeds within 8 h of sowing. These results further suggest that disease suppression in composts is related to microbial species that interact with the pathogen in its infection court and not in the bulk compost.

scholarly journals Microbial-Induced Carbon Competition in the Spermosphere Leads to Pathogen and Disease Suppression in a Municipal Biosolids Compost

2012 ◽  
Vol 102 (6) ◽  
pp. 588-596 ◽  
Author(s):  
Mei-Hsing Chen ◽  
Eric B. Nelson
Keyword(s):  
Linoleic Acid ◽  
Microbial Communities ◽  
Germ Tube ◽  
Disease Suppression ◽  
Cucumber Seed ◽  
Seed Exudate ◽  
Municipal Biosolids ◽  
Carbon Competition ◽  
Germ Tubes ◽  
Biosolids Compost

The aim of this study was to understand whether competition for fatty acids in plant seed exudates by compost-derived seed-colonizing microbial communities could explain the suppression of plant infections initiated by sporangia of Pythium ultimum. The germination behavior of P. ultimum sporangia in response to cucumber seeds was measured to determine the impact of seed-colonizing microbes on pathogen suppression. Seed-colonizing microbial communities from municipal biosolids compost utilized cucumber seed exudates and linoleic acid in vitro, reducing the respective stimulatory activity of these elicitors to P. ultimum sporangial germination. However, when sporangia were observed directly in the spermosphere of seeds sown in the compost medium, levels of germination and sporangial emptying did not differ from the responses in sand. The percentage of aborted germ tubes was greater after incubating sporangia in compost medium for 12-h than the level of germ tube abortion when sporangia were incubated in sand. Abortion did not occur if previously germinated sporangia were supplemented with cucumber seed exudate. Furthermore, removal of cucumber seed exudate after various stages of germ tube emergence resulted in an increase in aborted germ tubes over time. Adding increasing levels of glucose directly to the compost medium alleviated germ tube abortion in the spermosphere and also eliminated disease suppression. These data fail to support a role for linoleic acid competition in Pythium seedling disease suppression but provide evidence for general carbon competition mediated by seed-colonizing microbial communities as a mechanism for the suppression of Pythium seed infections in municipal biosolids compost.

scholarly journals Impact of Sacbrood Virus on Larval Microbiome of Apis mellifera and Apis cerana

Insects ◽  
2020 ◽  
Vol 11 (7) ◽  
pp. 439
Author(s):  
Rujipas Yongsawas ◽  
Veeranan Chaimanee ◽  
Jeffery S. Pettis ◽  
Humberto Freire Boncristiani Junior ◽  
Dawn Lopez ◽  
...  
Keyword(s):  
Apis Mellifera ◽  
Illumina Sequencing ◽  
Bacterial Communities ◽  
Apis Cerana ◽  
Rna Extraction ◽  
Treatment Groups ◽  
Sacbrood Virus ◽  
Polymerase Chain ◽  
The Impact ◽  
Food Mixture

In this study, we examined the impact of Sacbrood virus (SBV), the cause of larval honeybee (Apis mellifera) death, producing a liquefied a larva sac, on the gut bacterial communities on two larval honeybee species, Apis mellifera and Apis cerana. SBV was added into a worker jelly food mixture and bee larvae were grafted into each of the treatment groups for 24 h before DNA/RNA extraction. Confirmation of SBV infection was achieved using quantitative reverse transcription polymerase chain reaction (RT-qPCR) and visual symptomology. The 16S rDNA was sequenced by Illumina sequencing. The results showed the larvae were infected with SBV. The gut communities of infected A. cerana larvae exhibited a dramatic change compared with A. mellifera. In A. mellifera larvae, the Illumina sequencing revealed the proportion of Gilliamella, Snodgrassella and Fructobacillus was not significantly different, whereas in A. cerana, Gilliamella was significantly decreased (from 35.54% to 2.96%), however, with significant increase in Snodgrassella and Fructobacillus. The possibility of cross-infection should be further investigated.

scholarly journals Impact of Nosema Disease and American Foulbrood on Gut Bacterial Communities of Honeybees Apis mellifera

Insects ◽  
2021 ◽  
Vol 12 (6) ◽  
pp. 525
Author(s):  
Poonnawat Panjad ◽  
Rujipas Yongsawas ◽  
Chainarong Sinpoo ◽  
Chonthicha Pakwan ◽  
Phakamas Subta ◽  
...  
Keyword(s):  
Apis Mellifera ◽  
Bacterial Communities ◽  
454 Pyrosequencing ◽  
Gut Bacteria ◽  
Nosema Ceranae ◽  
Paenibacillus Larvae ◽  
American Foulbrood ◽  
Nosema Disease ◽  
The Impact ◽  
Pathogenic Infection

Honeybees, Apis mellifera, are important pollinators of many economically important crops. However, one of the reasons for their decline is pathogenic infection. Nosema disease and American foulbrood (AFB) disease are the most common bee pathogens that propagate in the gut of honeybees. This study investigated the impact of gut-propagating pathogens, including Nosema ceranae and Paenibacillus larvae, on bacterial communities in the gut of A. mellifera using 454-pyrosequencing. Pyrosequencing results showed that N. ceranae was implicated in the elimination of Serratia and the dramatic increase in Snodgrassella and Bartonella in adult bees’ guts, while bacterial communities of P. larvae-infected larvae were not affected by the infection. The results indicated that only N. ceranae had an impact on some core bacteria in the gut of A. mellifera through increasing core gut bacteria, therefore leading to the induction of dysbiosis in the bees’ gut.

scholarly journals The impact of RNA extraction method on accurate RNA sequencing from formalin-fixed paraffin-embedded tissues

BMC Cancer ◽  
2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Michal Marczyk ◽  
Chunxiao Fu ◽  
Rosanna Lau ◽  
Lili Du ◽  
Alexander J. Trevarton ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Gene Expression ◽  
Rna Sequencing ◽  
Rna Extraction ◽  
Formalin Fixed Paraffin ◽  
Ffpe Samples ◽  
Formalin Fixed Paraffin Embedded ◽  
The Impact ◽  
Formalin Fixed

Abstract Background Utilization of RNA sequencing methods to measure gene expression from archival formalin-fixed paraffin-embedded (FFPE) tumor samples in translational research and clinical trials requires reliable interpretation of the impact of pre-analytical variables on the data obtained, particularly the methods used to preserve samples and to purify RNA. Methods Matched tissue samples from 12 breast cancers were fresh frozen (FF) and preserved in RNAlater or fixed in formalin and processed as FFPE tissue. Total RNA was extracted and purified from FF samples using the Qiagen RNeasy kit, and in duplicate from FFPE tissue sections using three different kits (Norgen, Qiagen and Roche). All RNA samples underwent whole transcriptome RNA sequencing (wtRNAseq) and targeted RNA sequencing for 31 transcripts included in a signature of sensitivity to endocrine therapy. We assessed the effect of RNA extraction kit on the reliability of gene expression levels using linear mixed-effects model analysis, concordance correlation coefficient (CCC) and differential analysis. All protein-coding genes in the wtRNAseq and three gene expression signatures for breast cancer were assessed for concordance. Results Despite variable quality of the RNA extracted from FFPE samples by different kits, all had similar concordance of overall gene expression from wtRNAseq between matched FF and FFPE samples (median CCC 0.63–0.66) and between technical replicates (median expression difference 0.13–0.22). More than half of genes were differentially expressed between FF and FFPE, but with low fold change (median |LFC| 0.31–0.34). Two out of three breast cancer signatures studied were highly robust in all samples using any kit, whereas the third signature was similarly discordant irrespective of the kit used. The targeted RNAseq assay was concordant between FFPE and FF samples using any of the kits (CCC 0.91–0.96). Conclusions The selection of kit to purify RNA from FFPE did not influence the overall quality of results from wtRNAseq, thus variable reproducibility of gene signatures probably relates to the reliability of individual gene selected and possibly to the algorithm. Targeted RNAseq showed promising performance for clinical deployment of quantitative assays in breast cancer from FFPE samples, although numerical scores were not identical to those from wtRNAseq and would require calibration.

scholarly journals Impact of Intensive Land-Based Fish Culture in Qingdao, China, on the Bacterial Communities in Surrounding Marine Waters and Sediments

2011 ◽  
Vol 2011 ◽  
pp. 1-8 ◽  
Author(s):  
Qiufen Li ◽  
Yan Zhang ◽  
David Juck ◽  
Nathalie Fortin ◽  
Charles W. Greer
Keyword(s):  
Bacterial Communities ◽  
Gel Electrophoresis ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Fish Culture ◽  
Fish Ponds ◽  
Marine Area ◽  
Gradient Gel Electrophoresis ◽  
Reducing Bacteria ◽  
The Impact ◽  
Nitrate Reducing Bacteria

The impact of intensive land-based fish culture in Qingdao, China, on the bacterial communities in surrounding marine environment was analyzed. Culture-based studies showed that the highest counts of heterotrophic, ammonium-oxidizing, nitrifying, and nitrate-reducing bacteria were found in fish ponds and the effluent channel, with lower counts in the adjacent marine area and the lowest counts in the samples taken from 500 m off the effluent channel. Denaturing gradient gel electrophoresis (DGGE) analysis was used to assess total bacterial diversity. Fewer bands were observed from the samples taken from near the effluent channel compared with more distant sediment samples, suggesting that excess nutrients from the aquaculture facility may be reducing the diversity of bacterial communities in nearby sediments. Phylogenetic analysis of the sequenced DGGE bands indicated that the bacteria community of fish-culture-associated environments was mainly composed of Flavobacteriaceae, gamma- and deltaproteobacteria, including generaGelidibacter, Psychroserpen, Lacinutrix,andCroceimarina.

scholarly journals Alteration of Bacterial Communities in Anterior Nares and Skin Sites of Patients Undergoing Arthroplasty Surgery: Analysis by 16S rRNA and Staphylococcal-Specific tuf Gene Sequencing

2020 ◽  
Vol 8 (12) ◽  
pp. 1977
Author(s):  
Søren Iversen ◽  
Thor Bech Johannesen ◽  
Anna Cäcilia Ingham ◽  
Sofie Marie Edslev ◽  
Staffan Tevell ◽  
...  
Keyword(s):  
16S Rrna ◽  
Bacterial Communities ◽  
Gene Sequencing ◽  
Alpha Diversity ◽  
Clinical Samples ◽  
Staphylococcal Species ◽  
Tuf Gene ◽  
Staphylococcus Hominis ◽  
The Impact ◽  
Arthroplasty Surgery

The aim was to study alterations of bacterial communities in patients undergoing hip or knee arthroplasty to assess the impact of chlorhexidine gluconate soap decolonisation and systemic antibiotic prophylaxis. A Swedish multicentre, prospective collection of samples obtained from elective arthroplasty patients (n = 83) by swabbing anterior nares, skin sites in the groin and the site of planned surgery, before and after arthroplasty surgery, was analysed by 16S rRNA (V3-V4) gene sequencing and a complementary targeted tuf gene sequencing approach to comprehensively characterise alterations in staphylococcal communities. Significant reductions in alpha diversity was detected for both bacterial (p = 0.04) and staphylococcal (p = 0.03) groin communities after arthroplasty surgery with significant reductions in relative Corynebacterium (p = 0.001) abundance and Staphylococcus hominis (p = 0.01) relative staphylococcal abundance. In nares, significant reductions occurred for Staphylococcus hominis (p = 0.02), Staphylococcus haemolyticus (p = 0.02), and Staphylococcus pasteuri (p = 0.003) relative to other staphylococci. Staphylococcus aureus colonised 35% of anterior nares before and 26% after arthroplasty surgery. Staphylococcus epidermidis was the most abundant staphylococcal species at all sampling sites. No bacterial genus or staphylococcal species increased significantly after arthroplasty surgery. Application of a targeted tuf gene sequencing approach provided auxiliary staphylococcal community profiles and allowed species-level characterisation directly from low biomass clinical samples.

scholarly journals Characterisation of Bt maize IE09S034 in decomposition and response of soil bacterial communities

2021 ◽  
Author(s):  
Xiaoli Zhou ◽  
Jingang Liang ◽  
Ying Luan ◽  
Xinyuan Song ◽  
Zhengguang Zhang
Keyword(s):  
Metabolic Activity ◽  
Bacterial Communities ◽  
Bt Maize ◽  
Gm Crop ◽  
Soil Bacterial Communities ◽  
Significant Difference ◽  
Maize Residues ◽  
Soil Bacterial ◽  
June July ◽  
The Impact

Returning straw to the soil is an effective way to improve the soil quality. As genetically modified (GM) crops experience expanded growing scales, returning straw to the soil could also be necessary. However, the impact of GM crop straws on soil safety remains unclear. The environment (including soil types, humidity and temperature) can result in a significant difference in the diversity of soil bacterial communities. Here, we compared the impacts of the straw from Bt maize IE09S034 (IE) and near-isogenic non-Bt maize Zong31 (CK) on soil bacterial community and microbial metabolic activity in three different environments. Sampling was carried out following 6–10 months of decomposition (May, June, July, and August) in three localities in Chinese cities (Changchun, Jinan, and Beijing). Our results showed that Bt maize residues posed no direct impact on soil bacterial communities in contrast to the environment and decomposed time. The microbial functional diversity and metabolic activity showed no significant difference between IE and CK. The results could be a reference for further assessing the effect of Bt maize residues on the soil that promotes the commercialisation of Bt maize IE09S034.

scholarly journals Land use influences stream bacterial communities in lowland tropical watersheds

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Karina A. Chavarria ◽  
Kristin Saltonstall ◽  
Jorge Vinda ◽  
Jorge Batista ◽  
Megan Lindmark ◽  
...  
Keyword(s):  
Water Quality ◽  
Land Use ◽  
Bacterial Communities ◽  
Secondary Forest ◽  
Stream Water ◽  
Community Diversity ◽  
Forested Catchments ◽  
Unifrac Distance ◽  
Cattle Pasture ◽  
The Impact

AbstractLand use is known to affect water quality yet the impact it has on aquatic microbial communities in tropical systems is poorly understood. We used 16S metabarcoding to assess the impact of land use on bacterial communities in the water column of four streams in central Panama. Each stream was influenced by a common Neotropical land use: mature forest, secondary forest, silvopasture and traditional cattle pasture. Bacterial community diversity and composition were significantly influenced by nearby land uses. Streams bordered by forests had higher phylogenetic diversity (Faith’s PD) and similar community structure (based on weighted UniFrac distance), whereas the stream surrounded by traditional cattle pasture had lower diversity and unique bacterial communities. The silvopasture stream showed strong seasonal shifts, with communities similar to forested catchments during the wet seasons and cattle pasture during dry seasons. We demonstrate that natural forest regrowth and targeted management, such as maintaining and restoring riparian corridors, benefit stream-water microbiomes in tropical landscapes and can provide a rapid and efficient approach to balancing agricultural activities and water quality protection.

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