Optimization of an oxygen-based approach for community-level physiological profiling of soils
- Autores
- Zabaloy, Maria Celina; Lehman, R. Michael; Frey, Serita D.; Garland, Jay L.
- Año de publicación
- 2008
- Idioma
- inglés
- Tipo de recurso
- artículo
- Estado
- versión publicada
- Descripción
- Current approaches for rapid assessment of carbon source utilization by whole soil communities (i.e., community-level physiological profiling or CLPP) provide a limited, biased view of microbial communities with little connection to in situ activities. We developed an alternative CLPP approach based upon fluorometric detection of dissolved oxygen consumption in a microtiter platform which offers flexible manipulation of experimental factors. In the attempt to reduce oxygen re-dissolution, the wells were filled with liquid to very near the top and sealed. We found that filling the wells with 240 vs. 150 µl of sample improved the sensitivity of the system to discern both the response to a substrate amendment as low as 10 mg l-1 and un-amended, endogenous respiration. The preparation of a soil slurry facilitates inoculation into the microplate. Disruption of soil samples had a limited effect on the endogenous respiration in comparison to intact soil microbags in a 24-well microplate. Storage time (up to 33 days) reduced the level of activity in intact soil microbags but not in disrupted samples. A microcosm fertilization experiment was set to study the effects of N availability on the respiratory response in the plates. The use of soil organic carbon (SOC) and amended C-substrates (50 mg l-1) was increased by the addition of nitrogen (N) in the plate, and appeared N-limited shortly after microcosm fertilization. The addition of the eukaryotic inhibitor cycloheximide delayed the initial increase in fluorescence (time to minimum response) of several C sources (casein, acetate, asparagine, coumaric acid), varying among soils, which could be explained by the fungal use of these compounds. However, the extent of the inhibition caused by cycloheximide did not increase at higher fungal to bacteria ratios as estimated by PLFA analysis, indicating that the direct estimation of the fungal biomass from cycloheximide addition is not feasible. This paper provides an optimized, standardized protocol for soil analysis, and sets the basis for further validation studies that will continue to define the underlying capabilities/biases of this approach.
Fil: Zabaloy, Maria Celina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Centro de Recursos Naturales Renovables de la Zona Semiarida. Universidad Nacional del Sur. Centro de Recursos Naturales Renovables de la Zona Semiarida; Argentina
Fil: Lehman, R. Michael. North Central Agricultural Research Lab; Estados Unidos
Fil: Frey, Serita D.. University Of New Hampshire; Estados Unidos
Fil: Garland, Jay L.. Dynamac Corporation, Kennedy Space Center; Estados Unidos - Materia
-
Soil Testing
Bd Oxygen Biosensor System
Community Level Physiological Profiling
Soil Slurry
N Limitation
Fungal Respiration - Nivel de accesibilidad
- acceso abierto
- Condiciones de uso
- https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
- Repositorio
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/19976
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Optimization of an oxygen-based approach for community-level physiological profiling of soilsZabaloy, Maria CelinaLehman, R. MichaelFrey, Serita D.Garland, Jay L.Soil TestingBd Oxygen Biosensor SystemCommunity Level Physiological ProfilingSoil SlurryN LimitationFungal Respirationhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Current approaches for rapid assessment of carbon source utilization by whole soil communities (i.e., community-level physiological profiling or CLPP) provide a limited, biased view of microbial communities with little connection to in situ activities. We developed an alternative CLPP approach based upon fluorometric detection of dissolved oxygen consumption in a microtiter platform which offers flexible manipulation of experimental factors. In the attempt to reduce oxygen re-dissolution, the wells were filled with liquid to very near the top and sealed. We found that filling the wells with 240 vs. 150 µl of sample improved the sensitivity of the system to discern both the response to a substrate amendment as low as 10 mg l-1 and un-amended, endogenous respiration. The preparation of a soil slurry facilitates inoculation into the microplate. Disruption of soil samples had a limited effect on the endogenous respiration in comparison to intact soil microbags in a 24-well microplate. Storage time (up to 33 days) reduced the level of activity in intact soil microbags but not in disrupted samples. A microcosm fertilization experiment was set to study the effects of N availability on the respiratory response in the plates. The use of soil organic carbon (SOC) and amended C-substrates (50 mg l-1) was increased by the addition of nitrogen (N) in the plate, and appeared N-limited shortly after microcosm fertilization. The addition of the eukaryotic inhibitor cycloheximide delayed the initial increase in fluorescence (time to minimum response) of several C sources (casein, acetate, asparagine, coumaric acid), varying among soils, which could be explained by the fungal use of these compounds. However, the extent of the inhibition caused by cycloheximide did not increase at higher fungal to bacteria ratios as estimated by PLFA analysis, indicating that the direct estimation of the fungal biomass from cycloheximide addition is not feasible. This paper provides an optimized, standardized protocol for soil analysis, and sets the basis for further validation studies that will continue to define the underlying capabilities/biases of this approach.Fil: Zabaloy, Maria Celina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Centro de Recursos Naturales Renovables de la Zona Semiarida. Universidad Nacional del Sur. Centro de Recursos Naturales Renovables de la Zona Semiarida; ArgentinaFil: Lehman, R. Michael. North Central Agricultural Research Lab; Estados UnidosFil: Frey, Serita D.. University Of New Hampshire; Estados UnidosFil: Garland, Jay L.. Dynamac Corporation, Kennedy Space Center; Estados UnidosElsevier2008-12info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/19976Zabaloy, Maria Celina; Lehman, R. Michael; Frey, Serita D.; Garland, Jay L.; Optimization of an oxygen-based approach for community-level physiological profiling of soils; Elsevier; Soil Biology And Biochemistry; 40; 12; 12-2008; 2960-29690038-0717CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S0038071708002885info:eu-repo/semantics/altIdentifier/doi/10.1016/j.soilbio.2008.08.015info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:32:21Zoai:ri.conicet.gov.ar:11336/19976instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 10:32:22.132CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse |
dc.title.none.fl_str_mv |
Optimization of an oxygen-based approach for community-level physiological profiling of soils |
title |
Optimization of an oxygen-based approach for community-level physiological profiling of soils |
spellingShingle |
Optimization of an oxygen-based approach for community-level physiological profiling of soils Zabaloy, Maria Celina Soil Testing Bd Oxygen Biosensor System Community Level Physiological Profiling Soil Slurry N Limitation Fungal Respiration |
title_short |
Optimization of an oxygen-based approach for community-level physiological profiling of soils |
title_full |
Optimization of an oxygen-based approach for community-level physiological profiling of soils |
title_fullStr |
Optimization of an oxygen-based approach for community-level physiological profiling of soils |
title_full_unstemmed |
Optimization of an oxygen-based approach for community-level physiological profiling of soils |
title_sort |
Optimization of an oxygen-based approach for community-level physiological profiling of soils |
dc.creator.none.fl_str_mv |
Zabaloy, Maria Celina Lehman, R. Michael Frey, Serita D. Garland, Jay L. |
author |
Zabaloy, Maria Celina |
author_facet |
Zabaloy, Maria Celina Lehman, R. Michael Frey, Serita D. Garland, Jay L. |
author_role |
author |
author2 |
Lehman, R. Michael Frey, Serita D. Garland, Jay L. |
author2_role |
author author author |
dc.subject.none.fl_str_mv |
Soil Testing Bd Oxygen Biosensor System Community Level Physiological Profiling Soil Slurry N Limitation Fungal Respiration |
topic |
Soil Testing Bd Oxygen Biosensor System Community Level Physiological Profiling Soil Slurry N Limitation Fungal Respiration |
purl_subject.fl_str_mv |
https://purl.org/becyt/ford/1.6 https://purl.org/becyt/ford/1 |
dc.description.none.fl_txt_mv |
Current approaches for rapid assessment of carbon source utilization by whole soil communities (i.e., community-level physiological profiling or CLPP) provide a limited, biased view of microbial communities with little connection to in situ activities. We developed an alternative CLPP approach based upon fluorometric detection of dissolved oxygen consumption in a microtiter platform which offers flexible manipulation of experimental factors. In the attempt to reduce oxygen re-dissolution, the wells were filled with liquid to very near the top and sealed. We found that filling the wells with 240 vs. 150 µl of sample improved the sensitivity of the system to discern both the response to a substrate amendment as low as 10 mg l-1 and un-amended, endogenous respiration. The preparation of a soil slurry facilitates inoculation into the microplate. Disruption of soil samples had a limited effect on the endogenous respiration in comparison to intact soil microbags in a 24-well microplate. Storage time (up to 33 days) reduced the level of activity in intact soil microbags but not in disrupted samples. A microcosm fertilization experiment was set to study the effects of N availability on the respiratory response in the plates. The use of soil organic carbon (SOC) and amended C-substrates (50 mg l-1) was increased by the addition of nitrogen (N) in the plate, and appeared N-limited shortly after microcosm fertilization. The addition of the eukaryotic inhibitor cycloheximide delayed the initial increase in fluorescence (time to minimum response) of several C sources (casein, acetate, asparagine, coumaric acid), varying among soils, which could be explained by the fungal use of these compounds. However, the extent of the inhibition caused by cycloheximide did not increase at higher fungal to bacteria ratios as estimated by PLFA analysis, indicating that the direct estimation of the fungal biomass from cycloheximide addition is not feasible. This paper provides an optimized, standardized protocol for soil analysis, and sets the basis for further validation studies that will continue to define the underlying capabilities/biases of this approach. Fil: Zabaloy, Maria Celina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Centro de Recursos Naturales Renovables de la Zona Semiarida. Universidad Nacional del Sur. Centro de Recursos Naturales Renovables de la Zona Semiarida; Argentina Fil: Lehman, R. Michael. North Central Agricultural Research Lab; Estados Unidos Fil: Frey, Serita D.. University Of New Hampshire; Estados Unidos Fil: Garland, Jay L.. Dynamac Corporation, Kennedy Space Center; Estados Unidos |
description |
Current approaches for rapid assessment of carbon source utilization by whole soil communities (i.e., community-level physiological profiling or CLPP) provide a limited, biased view of microbial communities with little connection to in situ activities. We developed an alternative CLPP approach based upon fluorometric detection of dissolved oxygen consumption in a microtiter platform which offers flexible manipulation of experimental factors. In the attempt to reduce oxygen re-dissolution, the wells were filled with liquid to very near the top and sealed. We found that filling the wells with 240 vs. 150 µl of sample improved the sensitivity of the system to discern both the response to a substrate amendment as low as 10 mg l-1 and un-amended, endogenous respiration. The preparation of a soil slurry facilitates inoculation into the microplate. Disruption of soil samples had a limited effect on the endogenous respiration in comparison to intact soil microbags in a 24-well microplate. Storage time (up to 33 days) reduced the level of activity in intact soil microbags but not in disrupted samples. A microcosm fertilization experiment was set to study the effects of N availability on the respiratory response in the plates. The use of soil organic carbon (SOC) and amended C-substrates (50 mg l-1) was increased by the addition of nitrogen (N) in the plate, and appeared N-limited shortly after microcosm fertilization. The addition of the eukaryotic inhibitor cycloheximide delayed the initial increase in fluorescence (time to minimum response) of several C sources (casein, acetate, asparagine, coumaric acid), varying among soils, which could be explained by the fungal use of these compounds. However, the extent of the inhibition caused by cycloheximide did not increase at higher fungal to bacteria ratios as estimated by PLFA analysis, indicating that the direct estimation of the fungal biomass from cycloheximide addition is not feasible. This paper provides an optimized, standardized protocol for soil analysis, and sets the basis for further validation studies that will continue to define the underlying capabilities/biases of this approach. |
publishDate |
2008 |
dc.date.none.fl_str_mv |
2008-12 |
dc.type.none.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://purl.org/coar/resource_type/c_6501 info:ar-repo/semantics/articulo |
format |
article |
status_str |
publishedVersion |
dc.identifier.none.fl_str_mv |
http://hdl.handle.net/11336/19976 Zabaloy, Maria Celina; Lehman, R. Michael; Frey, Serita D.; Garland, Jay L.; Optimization of an oxygen-based approach for community-level physiological profiling of soils; Elsevier; Soil Biology And Biochemistry; 40; 12; 12-2008; 2960-2969 0038-0717 CONICET Digital CONICET |
url |
http://hdl.handle.net/11336/19976 |
identifier_str_mv |
Zabaloy, Maria Celina; Lehman, R. Michael; Frey, Serita D.; Garland, Jay L.; Optimization of an oxygen-based approach for community-level physiological profiling of soils; Elsevier; Soil Biology And Biochemistry; 40; 12; 12-2008; 2960-2969 0038-0717 CONICET Digital CONICET |
dc.language.none.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
info:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S0038071708002885 info:eu-repo/semantics/altIdentifier/doi/10.1016/j.soilbio.2008.08.015 |
dc.rights.none.fl_str_mv |
info:eu-repo/semantics/openAccess https://creativecommons.org/licenses/by-nc-sa/2.5/ar/ |
eu_rights_str_mv |
openAccess |
rights_invalid_str_mv |
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/ |
dc.format.none.fl_str_mv |
application/pdf application/pdf |
dc.publisher.none.fl_str_mv |
Elsevier |
publisher.none.fl_str_mv |
Elsevier |
dc.source.none.fl_str_mv |
reponame:CONICET Digital (CONICET) instname:Consejo Nacional de Investigaciones Científicas y Técnicas |
reponame_str |
CONICET Digital (CONICET) |
collection |
CONICET Digital (CONICET) |
instname_str |
Consejo Nacional de Investigaciones Científicas y Técnicas |
repository.name.fl_str_mv |
CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas |
repository.mail.fl_str_mv |
dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar |
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1844614337130397696 |
score |
13.070432 |