Effects of glyphosate on soil fungal communities: A field study / Efecto del glifosato en las comunidades fúngicas del suelo: estudio a campo

ABSTRACT The driving forces behind many soil processes are microorganisms and they are able to respond immediately to environmental changes. The soil microbial community impacts on many soil properties. More than one-third of the terrestrial ecosystems are semiarid. However, a limited number of studies have been conducted to characterize soil fungal communities in semiarid grasslands, in particular those of agricultural fields. The aim of this study was to explore changes in the diversity and structure of soil fungal communities in semiarid grasslands, after different doses of glyphosate were applied under field conditions. Changes in soil fungal communities were examined using different approaches including culturing, calcofluor white stain and denaturing gradient gel electrophoresis (DGGE). The different approaches complement each other, revealing different aspects of the effect of glyphosate on soil fungal communities. We demonstrated a negative effect of glyphosate on soil fungal biomass at high doses and an early and transitory stimulatory effect on soil fungal biomass. We also found a negative effect of glyphosate on the species richness of cultivable fungi and changes in the molecular structure of soil fungal communities after double doses or long-term glyphosate application. In summary, our findings demonstrate an overall negative effect of glyphosate on soil fungal communities.

RESUMEN Los microorganismos del suelo son los responsables de llevar a cabo la mayoría de los procesos biológicos que ocurren en el suelo, y son capaces de reaccionar ante el estrés ambiental. Más de un tercio de los ecosistemas terrestres son semiáridos. Sin embargo, son escasos los estudios realizados para caracterizar las comunidades fúngicas en suelos agrícolas en ecosistemas semiáridos. El objetivo del presente trabajo fue estudiar los cambios que se producen en la biomasa, la diversidad y la estructura de las comunidades fúngicas del suelo, luego de la aplicación de distintas dosis de glifosato en condiciones de campo. Se emplearon diferentes técnicas incluidas el cultivo, la tinción directa con blanco de calcoflúor y PCR acoplada a electroforesis en geles de gradiente desnaturalizante (DGGE). Las distintas metodologías empleadas se complementan entre sí al detectar cada una distintos aspectos del efecto del glifosato en las comunidades fúngicas del suelo. Se encontró que el glifosato produce un efecto negativo sobre la biomasa fúngica, también se encontró un efecto transitorio estimulante inmediatamente posterior a la aplicación del herbicida. Además, se vio un efecto negativo sobre la riqueza de hongos cultivables, así como también cambios en la estructura molecular de las comunidades luego de aplicaciones repetidas. En conclusión, se demostró un efecto negativo generalizado sobre las comunidades fúngicas del suelo.

Effects of glyphosate on soil fungal communities: A field study.

The driving forces behind many soil processes are microorganisms and they are able to respond immediately to environmental changes. The soil microbial community impacts on many soil properties. More than one-third of the terrestrial ecosystems are semiarid. However, a limited number of studies have been conducted to characterize soil fungal communities in semiarid grasslands, in particular those of agricultural fields. The aim of this study was to explore changes in the diversity and structure of soil fungal communities in semiarid grasslands, after different doses of glyphosate were applied under field conditions. Changes in soil fungal communities were examined using different approaches including culturing, calcofluor white stain and denaturing gradient gel electrophoresis (DGGE). The different approaches complement each other, revealing different aspects of the effect of glyphosate on soil fungal communities. We demonstrated a negative effect of glyphosate on soil fungal biomass at high doses and an early and transitory stimulatory effect on soil fungal biomass. We also found a negative effect of glyphosate on the species richness of cultivable fungi and changes in the molecular structure of soil fungal communities after double doses or long-term glyphosate application. In summary, our findings demonstrate an overall negative effect of glyphosate on soil fungal communities.

[Fungal biomass estimation in soils from southwestern Buenos Aires province (Argentina) using calcofluor white stain]. / Estimación de la biomasa fúngica en un suelo del sudoeste de la provincia de Buenos Aires (Argentina) con una tinción directa con blanco de calcoflúor.

Soil microorganisms are vital for ecosystem functioning because of the role they play in soil nutrient cycling. Agricultural practices and the intensification of land use have a negative effect on microbial activities and fungal biomass has been widely used as an indicator of soil health. The aim of this study was to analyze fungal biomass in soils from southwestern Buenos Aires province using direct fluorescent staining and to contribute to its use as an indicator of environmental changes in the ecosystem as well as to define its sensitivity to weather conditions. Soil samples were collected during two consecutive years. Soil smears were prepared and stained with two different concentrations of calcofluor, and the fungal biomass was estimated under an epifluorescence microscope. Soil fungal biomass varied between 2.23 and 26.89µg fungal C/g soil, being these values in the range expected for the studied soil type. The fungal biomass was positively related to temperature and precipitations. The methodology used was reliable, standardized and sensitive to weather conditions. The results of this study contribute information to evaluate fungal biomass in different soil types and support its use as an indicator of soil health for analyzing the impact of different agricultural practices.

Estimación de la biomasa fúngica en un suelo del sudoeste de la provincia de Buenos Aires (Argentina) con una tinción directa con blanco de calcoflúor / Fungal biomass estimation in soils from southwestern Buenos Aires province (Argentina) using calcofluor white stain

Los microorganismos del suelo son vitales para el correcto funcionamiento de los ecosistemas, principalmente por su papel en el ciclado de nutrientes. La intensificación del uso del suelo y las prácticas agrícolas alteran negativamente la actividad microbiana. La biomasa fúngica es uno de los parámetros más utilizados para estudiar el impacto de las actividades agrícolas en la estructura y el funcionamiento del suelo. El objetivo del presente trabajo fue estimar la biomasa fúngica en un suelo del sudoeste bonaerense con el fin de obtener valores de referencia que permitan usar este parámetro como un indicador de cambios en el ecosistema y, por otro lado, demostrar que la metodología empleada es sensible a las variaciones en las condiciones climáticas. Se colectaron muestras de suelos durante 2 años consecutivos. Se prepararon frotis de suelo y se tiñeron con soluciones de distintas concentraciones de blanco de calcoflúor y luego se estimó la biomasa fúngica observando los frotis con microscopio de epifluorescencia. Los valores de biomasa fúngica estimados variaron entre 2,23 y 26,89 μg Cfúngico/g de suelo y estuvieron dentro del rango esperable para el tipo de suelo estudiado. La biomasa fúngica mostró una relación positiva con la temperatura y las precipitaciones. La metodología empleada resultó ser confiable, repetible y sensible a cambios en las condiciones climáticas. Los resultados podrían usarse como valores de referencia para estudiar la biomasa fúngica de suelos bajo distintas condiciones y emplearse como indicadores del impacto de las distintas prácticas agrícolas sobre el ecosistema.

Soil microorganisms are vital for ecosystem functioning because of the role they play in soil nutrient cycling. Agricultural practices and the intensification of land use have a negative effect on microbial activities and fungal biomass has been widely used as an indicator of soil health. The aim of this study was to analyze fungal biomass in soils from southwestern Buenos Aires province using direct fluorescent staining and to contribute to its use as an indicator of environmental changes in the ecosystem as well as to define its sensitivity to weather conditions. Soil samples were collected during two consecutive years. Soil smears were prepared and stained with two different concentrations of calcofluor, and the fungal biomass was estimated under an epifluorescence microscope. Soil fungal biomass varied between 2.23 and 26.89 μg fungal C/g soil, being these values in the range expected for the studied soil type. The fungal biomass was positively related to temperature and precipitations. The methodology used was reliable, standardized and sensitive to weather conditions. The results of this study contribute information to evaluate fungal biomass in different soil types and support its use as an indicator of soil health for analyzing the impact of different agricultural practices.

Biomasa fúngica y bacteriana como indicadoras del secuestro de C en suelos de sabanas sustituidos por pinares en Uverito, Venezuela

Fungal and bacterial biomass as indicators of soil C sequestration in savannas soils substituted by pine plantations. A transformation of any natural ecosystem to an agricultural or forest system leads to an important soil modification, not only in the total carbon pool, but also in the carbon associated to the microbial biomass. This way, carbon quantification on soil quality is important for the determination of impacts of agricultural practices and land use changes. The aim of this study was to the determine, through the selective inhibition technique, the fungal and bacterial biomass, and fungal-to-bacterial ratio (F:B) in pine plantations (Pinus caribaea var. hondurensis), to establish if these parameters are sensible indicators of changes in the carbon content in Uverito soils (Venezuela). Furthermore, the inhibitor additivity ratio (IAR) and total combined inhibition (TCI) were carried out to determine if the antibiotics caused non-target inhibition. The quantification of fungal and bacterial biomass was carried out by using of cyloheximide as fungal inhibitor, and streptomycin and chloranphenicol as specific bacterial inhibitors. This research evidences that this land use change exerted a significant effect on soil microbial biomass, and shows that in pine plantations there is a dominance of the fungal component, in contrast to the native savanna, in which the bacterial biomass dominates. The substitution of native savanna by pine plantation in Uverito promotes a major soil carbon sequestration. The values of the inhibitor additivity ratio (IAR) as for native savanna as pine system, were both>1.0. The total combined inhibition (TCI) was smaller in the pine systems, from which it is possible to infer that a high proportion of microbial biomass was affected by the combination of the inhibitors. Rev. Biol. Trop. 58 (3): 977-989. Epub 2010 September 01.

Cualquier transformación de un ecosistema natural a un sistema agrícola o forestal conduce a una modificación importante no sólo del pool del carbono total, sino también del carbono asociado con la biomasa microbiana. Su cuantificación es importante en la determinación del impacto de las prácticas agrícolas y el cambio de uso de la tierra sobre la calidad del suelo. El objetivo de este estudio fue determinar, a través del método de inhibición selectiva, la biomasa fúngica y bacteriana y la relación (H:B) en suelos de sabana nativa sustituidos por pinares (Pinus caribaea var. hondurensis), para establecer si éstos parámetros son indicadores sensibles de cambios en el contenido de carbono en suelos de Uverito, Venezuela. La relación de aditividad del inhibidor (RAI) y la inhibición total por efecto combinado del inhibidor (ITC) se llevaron a cabo para determinar, si los inhibidores microbianos tuvieron actividad sobre otros organismos para los cuales éstos no estaban destinados. La cuantificación de la biomasa fúngica y bacteriana se llevó a cabo mediante el uso de la cycloheximida como inhibidor fúngico, y la estreptomicina y el cloranfenicol como inhibidores bacterianos. Esta investigación evidencia que este cambio de uso de la tierra ejerció un efecto significativo sobre la biomasa microbiana del suelo, y muestra que en el sistema de pinares existe una dominancia del componente fúngico, en contraste con la sabana nativa, en la cual domina la biomasa bacteriana. La sustitución de la sabana nativa por plantaciones de pino en Uverito, promueve un mayor secuestro del carbono en el suelo. Los valores de la relación de aditividad del inhibidor (RAI) tanto para la sabana nativa como para el sistema de pinares, resultaron ambos >1.0. La inhibición total combinada (ITC) resultó menor en el sistema de pinares; a partir de lo cual, es posible inferir que una elevada proporción de la biomasa microbiana fue afectada por la combinación de los inhibidores.