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Soil microbial dna extraction

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https://www.readbyqxmd.com/read/28330132/an-efficient-and-cost-effective-method-for-dna-extraction-from-athalassohaline-soil-using-a-newly-formulated-cell-extraction-buffer
#1
Avinash Narayan, Kunal Jain, Amita R Shah, Datta Madamwar
The present study describes the rapid and efficient indirect lysis method for environmental DNA extraction from athalassohaline soil by newly formulated cell extraction buffer. The available methods are mostly based on direct lysis which leads to DNA shearing and co-extraction of extra cellular DNA that influences the community and functional analysis. Moreover, during extraction of DNA by direct lysis from athalassohaline soil, it was observed that, upon addition of poly ethylene glycol (PEG), isopropanol or absolute ethanol for precipitation of DNA, salt precipitates out and affecting DNA yield significantly...
June 2016: 3 Biotech
https://www.readbyqxmd.com/read/28294483/the-detection-and-quantification-of-bacillus-thuringiensis-spores-from-soil-and-swabs-using-quantitative-pcr-as-a-model-system-for-routine-diagnostics-of-bacillus-anthracis
#2
Vera Sedlackova, Radka Dziedzinska, Vladimir Babak, Petr Kralik
AIMS: To optimize the DNA isolation for the routine detection and quantification of bacillary spores in soil and swabs. The procedure is primarily intended for diagnostics of Bacillus anthracis spores, but due to its high pathogenicity B. thuringiensis served as its surrogate organism. METHODS AND RESULTS: Various commercial kits for soils and swabs in combination with quantitative PCR were tested with different results. The PowerSoil DNA kit and the Ultra Clean Microbial DNA kit gave the best results for the extraction from soil and swabs, respectively...
March 10, 2017: Journal of Applied Microbiology
https://www.readbyqxmd.com/read/28217901/the-choice-of-the-dna-extraction-method-may-influence-the-outcome-of-the-soil-microbial-community-structure-analysis
#3
Sylwia Zielińska, Piotr Radkowski, Aleksandra Blendowska, Agnieszka Ludwig-Gałęzowska, Joanna M Łoś, Marcin Łoś
Metagenomics approaches and recent improvements in the next-generation sequencing methods, have become a method of choice in establishing a microbial population structure. Many commercial soil DNA extraction kits are available and due to their efficiency they are replacing traditional extraction protocols. However, differences in the physicochemical properties of soil samples require optimization of DNA extraction techniques for each sample separately. The aim of this study was to compare the efficiency, quality, and diversity of genetic material extracted with the use of commonly used kits...
February 20, 2017: MicrobiologyOpen
https://www.readbyqxmd.com/read/28203533/modification-of-extraction-method-for-community-dna-isolation-from-salt-affected-compact-wasteland-soil-samples
#4
Purvi Zaveri, Rushika Patel, Meghavi Patel, Devki Sarodia, Nasreen S Munshi
To overcome the issue of interferences by salt and compactness in release of bacterial cell required for lysis, method described by Yeates et al. (1998), was optimized for isolation of genomic material (Deoxyribo Nucleic Acid, DNA) from soil microbial community by addition of Al(NH4)SO4. Very low total viable count was observed in the samples tested and hence use of higher amount of soil is required primarily for DNA isolation from wasteland soils. The method proves itself efficient where commercially available bead beating and enzymatic lysis methods could not give isolation of any amount of community genomic DNA due to compact nature and salt concentrations present in soil...
2017: MethodsX
https://www.readbyqxmd.com/read/28168105/successive-dna-extractions-improve-characterization-of-soil-microbial-communities
#5
Mauricio R Dimitrov, Annelies J Veraart, Mattias de Hollander, Hauke Smidt, Johannes A van Veen, Eiko E Kuramae
Currently, characterization of soil microbial communities relies heavily on the use of molecular approaches. Independently of the approach used, soil DNA extraction is a crucial step, and success of downstream procedures will depend on how well DNA extraction was performed. Often, studies describing and comparing soil microbial communities are based on a single DNA extraction, which may not lead to a representative recovery of DNA from all organisms present in the soil. The use of successive DNA extractions might improve soil microbial characterization, but the benefit of this approach has only been limitedly studied...
2017: PeerJ
https://www.readbyqxmd.com/read/28093673/metagenomic-insights-into-effects-of-spent-engine-oil-perturbation-on-the-microbial-community-composition-and-function-in-a-tropical-agricultural-soil
#6
Lateef B Salam, Sunday O Obayori, Francisca O Nwaokorie, Aisha Suleiman, Raheemat Mustapha
Analyzing the microbial community structure and functions become imperative for ecological processes. To understand the impact of spent engine oil (SEO) contamination on microbial community structure of an agricultural soil, soil microcosms designated 1S (agricultural soil) and AB1 (agricultural soil polluted with SEO) were set up. Metagenomic DNA extracted from the soil microcosms and sequenced using Miseq Illumina sequencing were analyzed for their taxonomic and functional properties. Taxonomic profiling of the two microcosms by MG-RAST revealed the dominance of Actinobacteria (23...
January 16, 2017: Environmental Science and Pollution Research International
https://www.readbyqxmd.com/read/28074923/bacterial-community-composition-in-costal-dunes-of-the-mediterranean-along-a-gradient-from-the-sea-shore-to-the-inland
#7
Haggai Wasserstrom, Susanne Kublik, Rachel Wasserstrom, Stefanie Schulz, Michael Schloter, Yosef Steinberger
Sand dunes are unique ecosystems with distinct features which limited the accumulation of biomass. The distance from seashore affects both the physical properties of the sand dunes and the biota living above- and below ground. The goal of the present study was to determine the effects of the distance from shore to inland on soil bacterial community composition during wet and dry season. We studied a chronosequence of sites close to the eastern Mediterranean coast. Bacterial diversity was assessed using directly extracted DNA from soil samples and 16 S ribosomal RNA gene fingerprinting...
January 11, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28050848/microbial-communities-in-pyrene-amended-soil-compost-mixture-and-fertilized-soil
#8
Iris K U Adam, Márcia Duarte, Jananan Pathmanathan, Anja Miltner, Thomas Brüls, Matthias Kästner
Polycyclic aromatic hydrocarbons are distributed ubiquitously in the environment and form metabolites toxic to most organisms. Organic amendment of PAH contaminated soil with compost and farmyard manure has proven to be efficient for PAH bioremediation mediated by native microorganisms, even though information on the identity of PAH degraders in organic-amended soil is still scarce. Here we provide molecular insight into the bacterial communities in soil amended with compost or farmyard manure for which the degradation mass balances of (13)C-labeled pyrene have been recently published and assess the relevant bacterial genera capable of degrading pyrene as a model PAH...
December 2017: AMB Express
https://www.readbyqxmd.com/read/28028549/ssunique-detecting-sequence-novelty-in-microbiome-surveys
#9
Michael D J Lynch, Josh D Neufeld
High-throughput sequencing of small-subunit (SSU) rRNA genes has revolutionized understanding of microbial communities and facilitated investigations into ecological dynamics at unprecedented scales. Such extensive SSU rRNA gene sequence libraries, constructed from DNA extracts of environmental or host-associated samples, often contain a substantial proportion of unclassified sequences, many representing organisms with novel taxonomy (taxonomic "blind spots") and potentially unique ecology. Indeed, these novel taxonomic lineages are associated with so-called microbial "dark matter," which is the genomic potential of these lineages...
November 2016: MSystems
https://www.readbyqxmd.com/read/27660624/the-willow-microbiome-is-influenced-by-soil-petroleum-hydrocarbon-concentration-with-plant-compartment-specific-effects
#10
Stacie Tardif, Étienne Yergeau, Julien Tremblay, Pierre Legendre, Lyle G Whyte, Charles W Greer
The interaction between plants and microorganisms, which is the driving force behind the decontamination of petroleum hydrocarbon (PHC) contamination in phytoremediation technology, is poorly understood. Here, we aimed at characterizing the variations between plant compartments in the microbiome of two willow cultivars growing in contaminated soils. A field experiment was set-up at a former petrochemical plant in Canada and after two growing seasons, bulk soil, rhizosphere soil, roots, and stems samples of two willow cultivars (Salix purpurea cv...
2016: Frontiers in Microbiology
https://www.readbyqxmd.com/read/27627898/modified-method-for-the%C3%A2-extraction-of-mrna-from-paddy-soils
#11
Hongling Qin, Xiangbi Chen, Yafang Tang, Haijun Hou, Rong Sheng, Jinlin Shen
OBJECTIVE: Paddy soils contain relatively greater organic matter and water contents than other soils thereby limiting effective mRNA extraction. A modification of the conventional mRNA soil extraction method specific to paddy soils is described. RESULTS: Two main modifications for co-extraction of DNA and RNA are: (1) addition of 20 % (w/v) sodium dodecyl sulphate to 10 % (w/v) hexadecyltrimethylammonium bromide extraction buffer, and (2) fresh soil, initially frozen at -80 °C, is immediately immersed in extraction buffer...
December 2016: Biotechnology Letters
https://www.readbyqxmd.com/read/27580466/solanum-lycopersicum-tomato-hosts-robust-phyllosphere-and-rhizosphere-bacterial-communities-when-grown-in-soil-amended-with-various-organic-and-synthetic-fertilizers
#12
Sarah M Allard, Christopher S Walsh, Anna E Wallis, Andrea R Ottesen, Eric W Brown, Shirley A Micallef
Due to the intimate association between plants and their microbial symbionts, an examination of the influence of agricultural practices on phytobiome structure and diversity could foster a more comprehensive understanding of plant health and produce safety. Indeed, the impact of upstream crop producti006Fn practices cannot be overstated in their role in assuring an abundant and safe food supply. To assess whether fertilizer type impacted rhizosphere and phyllosphere bacterial communities associating with tomato plants, the bacterial microbiome of tomato cv...
December 15, 2016: Science of the Total Environment
https://www.readbyqxmd.com/read/27528083/soil-pretreatment-and-fast-cell-lysis-for-direct-polymerase-chain-reaction-from-forest-soils-for-terminal-restriction-fragment-length-polymorphism-analysis-of-fungal-communities
#13
Fei Cheng, Lin Hou, Keith Woeste, Zhengchun Shang, Xiaobang Peng, Peng Zhao, Shuoxin Zhang
Humic substances in soil DNA samples can influence the assessment of microbial diversity and community composition. Using multiple steps during or after cell lysis adds expenses, is time-consuming, and causes DNA loss. A pretreatment of soil samples and a single step DNA extraction may improve experimental results. In order to optimize a protocol for obtaining high purity DNA from soil microbiota, five prewashing agents were compared in terms of their efficiency and effectiveness in removing soil contaminants...
October 2016: Brazilian Journal of Microbiology: [publication of the Brazilian Society for Microbiology]
https://www.readbyqxmd.com/read/27524249/potential-risks-of-the-residue-from-samarco-s-mine-dam-burst-bento-rodrigues-brazil
#14
Fabiana Roberta Segura, Emilene Arusievicz Nunes, Fernanda Pollo Paniz, Ana Carolina Cavalheiro Paulelli, Gabriela Braga Rodrigues, Gilberto Úbida Leite Braga, Walter Dos Reis Pedreira Filho, Fernando Barbosa, Giselle Cerchiaro, Fábio Ferreira Silva, Bruno Lemos Batista
On November 5th, 2015, Samarco's iron mine dam - called Fundão - spilled 50-60 million m(3) of mud into Gualaxo do Norte, a river that belongs to Rio Doce Basin. Approximately 15 km(2) were flooded along the rivers Gualaxo do Norte, Carmo and Doce, reaching the Atlantic Ocean on November 22nd, 2015. Six days after, our group collected mud, soil and water samples in Bento Rodrigues (Minas Gerais, Brazil), which was the first impacted area. Overall, the results, water samples - potable and surface water from river - presented chemical elements concentration according to Brazilian environmental legislations, except silver concentration in surface water that ranged from 1...
November 2016: Environmental Pollution
https://www.readbyqxmd.com/read/27454095/ecological-restoration-of-a-copper-polluted-vineyard-long-term-impact-of-farmland-abandonment-on-soil-bio-chemical-properties-and-microbial-communities
#15
Luciano Cavani, Luisa M Manici, Francesco Caputo, Elisabetta Peruzzi, Claudio Ciavatta
This study aimed at investigating the degree of interference of high soil copper (Cu) contamination when an old vineyard is converted into a protected area. This study was performed within an intensive agricultural system; it was organized into a two-factorial nested design to analyze the impact of management (conventional vs re-naturalized orchard) and position within each orchard (tree-rows and strips). Chemical and biochemical properties along with bacterial and fungal communities, evaluated with PCR-DGGE starting from total soil DNA, were analyzed...
November 1, 2016: Journal of Environmental Management
https://www.readbyqxmd.com/read/27450478/desert-perennial-shrubs-shape-the-microbial-community-miscellany-in-laimosphere-and-phyllosphere-space
#16
Varsik Martirosyan, Adrian Unc, Gad Miller, Tirza Doniger, Chaim Wachtel, Yosef Steinberger
Microbial function, composition, and distribution play a fundamental role in ecosystem ecology. The interaction between desert plants and their associated microbes is expected to greatly affect their response to changes in this harsh environment. Using comparative analyses, we studied the impact of three desert shrubs, Atriplex halimus (A), Artemisia herba-alba (AHA), and Hammada scoparia (HS), on soil- and leaf-associated microbial communities. DNA extracted from the leaf surface and soil samples collected beneath the shrubs were used to study associated microbial diversity using a sequencing survey of variable regions of bacterial 16S rRNA and fungal ribosomal internal transcribed spacer (ITS1)...
October 2016: Microbial Ecology
https://www.readbyqxmd.com/read/27403572/removal-of-exogenous-materials-from-the-outer-portion-of-frozen-cores-to-investigate-the-ancient-biological-communities-harbored-inside
#17
Robyn A Barbato, Natàlia Garcia-Reyero, Karen Foley, Robert Jones, Zoe Courville, Thomas Douglas, Edward Perkins, Charles M Reynolds
The cryosphere offers access to preserved organisms that persisted under past environmental conditions. In fact, these frozen materials could reflect conditions over vast time periods and investigation of biological materials harbored inside could provide insight of ancient environments. To appropriately analyze these ecosystems and extract meaningful biological information from frozen soils and ice, proper collection and processing of the frozen samples is necessary. This is especially critical for microbial and DNA analyses since the communities present may be so uniquely different from modern ones...
2016: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/27387909/active-microorganisms-in-forest-soils-differ-from-the-total-community-yet-are-shaped-by-the-same-environmental-factors-the-influence-of-ph-and-soil-moisture
#18
Karl J Romanowicz, Zachary B Freedman, Rima A Upchurch, William A Argiroff, Donald R Zak
Predicting the impact of environmental change on soil microbial functions requires an understanding of how environmental factors shape microbial composition. Here, we investigated the influence of environmental factors on bacterial and fungal communities across an expanse of northern hardwood forest in Michigan, USA, which spans a 500-km regional climate gradient. We quantified soil microbial community composition using high-throughput DNA sequencing on coextracted rDNA (i.e. total community) and rRNA (i.e...
October 2016: FEMS Microbiology Ecology
https://www.readbyqxmd.com/read/27313569/size-matters-assessing-optimum-soil-sample-size-for-fungal-and-bacterial-community-structure-analyses-using-high-throughput-sequencing-of-rrna-gene-amplicons
#19
C Ryan Penton, Vadakattu V S R Gupta, Julian Yu, James M Tiedje
We examined the effect of different soil sample sizes obtained from an agricultural field, under a single cropping system uniform in soil properties and aboveground crop responses, on bacterial and fungal community structure and microbial diversity indices. DNA extracted from soil sample sizes of 0.25, 1, 5, and 10 g using MoBIO kits and from 10 and 100 g sizes using a bead-beating method (SARDI) were used as templates for high-throughput sequencing of 16S and 28S rRNA gene amplicons for bacteria and fungi, respectively, on the Illumina MiSeq and Roche 454 platforms...
2016: Frontiers in Microbiology
https://www.readbyqxmd.com/read/27145490/using-geochemical-indicators-to-distinguish-high-biogeochemical-activity-in-floodplain-soils-and-sediments
#20
Amy Kenwell, Alexis Navarre-Sitchler, Rodrigo Prugue, John R Spear, Amanda S Hering, Reed M Maxwell, Rosemary W H Carroll, Kenneth H Williams
A better understanding of how microbial communities interact with their surroundings in physically and chemically heterogeneous subsurface environments will lead to improved quantification of biogeochemical reactions and associated nutrient cycling. This study develops a methodology to predict potential elevated rates of biogeochemical activity (microbial "hotspots") in subsurface environments by correlating microbial DNA and aspects of the community structure with the spatial distribution of geochemical indicators in subsurface sediments...
September 1, 2016: Science of the Total Environment
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