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1.
Appl Microbiol Biotechnol ; 98(14): 6467-85, 2014.
Article in English | MEDLINE | ID: mdl-24797311

ABSTRACT

Plastics are used widely as agricultural mulches to suppress weeds and retain soil moisture. Disposal of conventional plastic mulches requires physical removal for disposal in a landfill or incineration. Biodegradable plastic mulches that could be tilled into the soil at the end of a growing season represent an attractive alternative to conventional plastic mulches. In this study, three commercially available mulches labeled as "biodegradable" and one experimental, potentially biodegradable mulch were used during a tomato growing season, and then buried in field soil at three locations for approximately 6 months, as would occur typically in an agricultural setting. Degradation after 6 months in soil was minimal for all but the cellulosic mulch. After removal of mulches from soil, fungi were isolated from the mulch surfaces and tested for their ability to colonize and degrade the same mulches in pure culture. The majority of culturable soil fungi that colonized biodegradable mulches were within the family Trichocomaceae (which includes beneficial, pathogenic, and mycotoxigenic species of Aspergillus and Penicillium). These isolates were phylogenetically similar to fungi previously reported to degrade both conventional and biodegradable plastics. Under pure culture conditions, only a subset of fungal isolates achieved detectable mulch degradation. No isolate substantially degraded any mulch. Additionally, DNA was extracted from bulk soil surrounding buried mulches and ribosomal DNA was used to assess the soil microbial community. Soil microbial community structure was significantly affected by geographical location, but not by mulch treatments.


Subject(s)
Biodegradable Plastics , Biota , Fungi/isolation & purification , Fungi/metabolism , Soil Microbiology , Agriculture/methods , Fungi/classification , Fungi/growth & development , Molecular Sequence Data , Sequence Analysis, DNA
2.
J Vis Exp ; (75): e50373, 2013 May 10.
Article in English | MEDLINE | ID: mdl-23712218

ABSTRACT

Fungi native to agricultural soils that colonized commercially available biodegradable mulch (BDM) films were isolated and assessed for potential to degrade plastics. Typically, when formulations of plastics are known and a source of the feedstock is available, powdered plastic can be suspended in agar-based media and degradation determined by visualization of clearing zones. However, this approach poorly mimics in situ degradation of BDMs. First, BDMs are not dispersed as small particles throughout the soil matrix. Secondly, BDMs are not sold commercially as pure polymers, but rather as films containing additives (e.g. fillers, plasticizers and dyes) that may affect microbial growth. The procedures described herein were used for isolates acquired from soil-buried mulch films. Fungal isolates acquired from excavated BDMs were tested individually for growth on pieces of new, disinfested BDMs laid atop defined medium containing no carbon source except agar. Isolates that grew on BDMs were further tested in liquid medium where BDMs were the sole added carbon source. After approximately ten weeks, fungal colonization and BDM degradation were assessed by scanning electron microscopy. Isolates were identified via analysis of ribosomal RNA gene sequences. This report describes methods for fungal isolation, but bacteria also were isolated using these methods by substituting media appropriate for bacteria. Our methodology should prove useful for studies investigating breakdown of intact plastic films or products for which plastic feedstocks are either unknown or not available. However our approach does not provide a quantitative method for comparing rates of BDM degradation.


Subject(s)
Agriculture/methods , Bacteria/metabolism , Biodegradable Plastics/metabolism , Fungi/metabolism , Soil Microbiology , Biodegradable Plastics/chemistry , Biodegradation, Environmental
3.
Microb Ecol ; 55(3): 500-11, 2008 Apr.
Article in English | MEDLINE | ID: mdl-17786504

ABSTRACT

The impact and frequency of forest harvesting could significantly affect soil microbial community (SMC) structure and functioning. The ability of soil microorganisms to perform biogeochemical processes is critical for sustaining forest productivity and has a direct impact on decomposition dynamics and carbon storage potential. The Wind River Canopy Crane Research Forest in SW, WA, provided a unique opportunity to study a forest chronosequence and the residual effects of harvesting on the SMC in comparison to old-growth forests. The objective of this study was to determine the effect of clear-cutting and stand age on temporal dynamics of SMC and physiological stress markers using phospholipid fatty acid (PLFA) profiling. Soil microbial PLFA profiles were determined seven times over 22 months (Nov. 02 to Sep. 04) in old-growth coniferous forest stands (300-500 years) and 8 (CC8)- or 25 (CC25)-year-old replanted clear-cuts. PLFA patterns of the SMC shifted because of clear-cutting, but seasonal temporal changes had greater shifts than differences among stand age. The microbial biomass (total PLFA) and bacterial, fungal, and selected other PLFAs were significantly reduced in CC8 but not in CC25 sites relative to the old-growth sites. An increase in stress indicators [PLFA ratios of saturated/monsaturated and (cy17:0 + cy19:0)/(16:1omega7 + 18:1omega7)] in late summer was related to water stress. Although the canopy and litter input are quite different for a 25-year clear-cut compared to virgin old-growth forest, we conclude that the composition of the microbial communities, 25 years after clear-cutting, has recovered sufficiently to be much more similar to old-growth forests than a recent clear-cut at this Pacific Northwest forest site. The study shows the potential of PLFA analysis for profiling microbial communities and their stress status under field conditions, but wide temporal shifts emphasize the need for sampling over seasons to fully interpret ecosystem management impacts on microbial populations.


Subject(s)
Fatty Acids/analysis , Phospholipids/analysis , Pseudotsuga/microbiology , Seasons , Soil Microbiology , Soil/analysis , Animals , Bacteria/metabolism , Biomass , Eukaryota/metabolism , Fatty Acids/isolation & purification , Fungi/metabolism , Phospholipids/isolation & purification , Pseudotsuga/growth & development , Temperature , Time Factors , Trees , Washington , Water/analysis
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