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Abstract

The effects of prescribed burning of bushes on the soil are evaluated in the short term, at 5 and 9 years, using glomalin fractions as indicators. This evaluation is carried out for three soil thicknesses (0-1, 1-2 and 2-3 cm) and in quadruplicate at each sampling time. The glomalin values (total, residual and labile), although higher, in absolute value, in the shallowest cm of soil, rarely show significant differences in depth. Immediately after burning, soils double their total and residual glomalin content with respect to control or unburned soils; these values decrease with time. Labile glomalin, which is not significantly affected immediately after burning, is reduced by half after a few years. In short, a series of changes in the different glomalin fractions are identified, despite the low severity of this type of burning.
477
Revista de Ciências Agrárias
,
2022,
45(4): 477-481
Evolución de la glomalina tras la quema prescrita
de matorral en suelos de alta montaña
(Pirineo Central, NE-España)
Glomalin-related soil protein evolution after prescribed burning
of scrub in mountain soils (Central Pyrenees, NE-Spain)
Andoni Alfaro-Leranoz1,*, Mohamed Emran1,2 & David Badía-Villas1
1 GEOFOREST, Departamento de Ciencias Agrarias y del Medio Natural, Instituto de Investigación en Ciencias Ambientales (IUCA), Escuela Politécnica Superior de Huesca, Universidad de
Zaragoza, 22071 Huesca, España
2 Land and Water Technologies Department, Arid Lands Cultivation Research Institute, City of Scientic Research and Technological Applications (SRTA-City), 21934 New Borg El-Arab City,
Alexandria, Egypt
(*E-mail: a.alfaroler@unizar.es)
https://doi.org/10.19084/rca.28572
RESUMEN
Se evalúan los efectos de la quema prescrita de matorral sobre el suelo a corto plazo, a los 5 y a los 9 años usando como
indicadores las fracciones de glomalina. Dicha evaluación se efectúa para tres espesores (0-1, 1-2 y 2-3 cm) de suelo y por
cuatriplicado en cada momento de muestreo. Los valores de glomalina (total, residual y lábil) aunque son más elevadas,
en valor absoluto, en el cm del suelo más supercial, raramente muestra diferencias signicativas en profundidad.
Inmediatamente tras la quema, los suelos duplican su contenido en glomalina total y residual con respecto a los suelos
control o no quemados; dichos valores disminuyen con el tiempo. La glomalina lábil, que no se ve signicativamente
afectada justo tras la quema, se reduce a la mitad pasados los años. En denitiva, se identican una serie de cambios en
las diferentes fracciones de glomalina, a pesar de la baja severidad de este tipo de quemas.
Palabras clave: Quema prescrita; Proteínas del suelo relacionadas con la glomalina.
ABSTRACT
The eects of prescribed burning of bushes on the soil are evaluated in the short term, at 5 and 9 years, using
glomalin fractions as indicators. This evaluation is carried out for three soil thicknesses (0-1, 1-2 and 2-3 cm) and in
quadruplicate at each sampling time. The glomalin values (total, residual and labile), although higher, in absolute value,
in the shallowest cm of soil, rarely show signicant dierences in depth. Immediately after burning, soils double their
total and residual glomalin content with respect to control or unburned soils; these values decrease with time. Labile
glomalin, which is not signicantly aected immediately after burning, is reduced by half after a few years. In short,
a series of changes in the dierent glomalin fractions are identied, despite the low severity of this type of burning.
Keywords: Prescribed re, Glomalin-related soil protein.
478 Revista de Ciências Agrárias
,
2022,
45(4): 477-481
INTRODUCCIÓN
Se denomina glomalina a un conjunto de pequeñas
glicoproteínas que se encuentran en abundancia
en las esporas e hifas de los hongos micorrízicos
arbusculares; por ello, se suele hablar proteínas
del suelo relacionadas con la glomalina (PSRG). Su
presencia se relaciona con la agregación del suelo
(Hontoria et al., 2009; Emran et al., 2012). Una gran
parte de la glomalina es de cierta recalcitrancia,
por lo que interviene en el secuestro de carbono
orgánico y se utiliza como indicador de la calidad
del suelo (Rillig et al., 2004; Emran et al., 2020; 2021).
La cantidad total de glomalina y sus fracciones se
usan como indicadores de perturbaciones en el
suelo como el laboreo intenso o los incendios (Lo-
zano et al., 2018; Shari et al., 2018).
Por otro lado, las quemas prescritas de matorral en
alta montaña tratan de recuperar zonas de pastos
de verano para el ganado trashumante sin gene-
ran daños en el suelo. Sin embargo, las propieda-
des edácas más sensibles, como las biológicas,
pueden verse afectadas por dichas quemas (Giro-
na-García et al., 2018). Por ello, en este trabajo se
usa la glomalina, y sus fracciones, como indicador
de cambios en suelos sometidos a quemas prescri-
tas de matorral.
MATERIAL Y MÉTODOS
Con el objetivo de valorar si el fuego afecta al suelo
se muestrearon zonas control y zonas de matorral
recién quemadas (Q0), a medio (Q5, 5 años) y a lar-
go plazo (Q9, 9 años), todas ellas por cuatriplicado
(n=4). Se muestrearon suelos a diversas profundi-
dades (0-1, 1-2 y 2-3 cm) para detectar qué espesor
podía verse afectado por la quema.
Las concentraciones de glomalina (total, residual
y fácilmente extraíble o lábil) se determinaron en
muestras de suelo, tamizado a 2 mm, según lo
descrito por Wright y Upadhyaya (1996). La frac-
ción total de la glomalina se extrajo con citrato
de sodio 50 mM, pH 8.0, en ciclos secuenciales de
autoclavado a 121ºC (250 ºF) de 60 minutos, hasta
que el sobrenadante no mostró el color caracte-
rístico de la glomalina. La glomalina fácilmente
extraíble o lábil, se extrajo usando citrato sódico
20 mM a pH 7.0, tras un solo ciclo de autoclavado
a 121 ºC de 30 minutos (Wright y Upadhyaya,
1996).
Se obtuvo la glomalina residual por diferencia en-
tre la glomalina total y la lábil. La concentración de
glomalina en los extractos se determinó mediante
el ensayo de proteínas de Bradford, con albúmina
de suero bovino como estándar.
Adicionalmente se determinó el carbono orgánico
por calcinación de la materia orgánica y aplica-
ción del factor de Van Bemmelen, el ujo de CO2
por captura en sosa, el C microbiano por fumiga-
ción-extracción y la estabilidad de los agregados
(1-2 mm) por tamizado en húmedo (Alfaro-Lera-
noz et al., 2022).
El análisis estadístico se realizó con el progra-
ma STATISTICA 10 de StatSoft, Inc. Se realizó un
ANOVA de medidas repetidas para analizar la va-
riabilidad de las fracciones de glomalina en fun-
ción de los dos factores categóricos: tiempo desde
la quema prescrita y profundidad del suelo. Tam-
bién se aplicó la prueba HSD de Tukey de compa-
ración de medias.
Área de estudio
La zona de estudio (Asín de Broto) se encuentra en
el piso subalpino del Pirineo Central, en el Noreste
de España (Tabla 1).
Se trata de una zona de pastos de verano que, ante
el progresivo descenso de la carga ganadera de las
últimas décadas, se ha visto matorralizada por eri-
zón (Echinospartium horridum (Vahl) Rothm), comu-
nidad vegetal con muy pocas especies (n=6 ± 3) y,
Tabla 1 - Características de la zona de estudio
Asín de Broto
Coordenadas 42º 31’ 12.3” N
0º 06’ 02.4” O
Altitud (m) 1650
Orientación Oeste
Pendiente (%) 35
Temperatura media anual (ºC) 8.8
Precipitación (mm/año) 1120
Suelo (WRB) Calcaric Cambisol
479
Alfaro-Leranoz et al., Evolución de la glomalina tras la quema prescrita
por tanto, muy baja diversidad (índice de Shannon
de 0,858 ± 0,467) y bajo valor pastoral (Badía-Villas
et al., 2017). Las quemas prescritas de este matorral
espinoso las realizaron los Equipos de Prevención
Integral de Incendios Forestales (EPRIF), en épo-
ca invernal (Tabla 2), realizándose el muestreo de
suelos simultáneamente en todas ellas en marzo
del año 2021.
RESULTADOS
Los valores de glomalina (total, residual y lábil) son
más elevados, en valor absoluto, en el cm del suelo
más supercial y descienden progresivamente en
profundidad, si bien las diferencias son raramen-
te signicativas (Tabla 3). Inmediatamente tras la
quema (Q0), los suelos duplican su contenido en
glomalina total y residual con respecto a los suelos
control o no quemados (C). Con el tiempo, a los 5
años (Q5), y especialmente a los 9 años (Q9) tras la
quema, esos valores disminuyen (Tabla 3). Por otro
lado, la glomalina lábil no se ve signicativamen-
te afectada tras la quema (Q0) pero se reduce a la
mitad posteriormente (Q5 y Q9 años), a pesar de
la recuperación de la cubierta vegetal (Badía-Villas
et al., 2017).
Existe una correlación positiva y signicativa
(p<0,01) del C orgánico oxidable del suelo con las
tres fracciones de glomalina estudiadas, aspecto
contrastado en estudios previos (Emran et al., 2012;
Lozano et al., 2016; Shari et al., 2018). Se ha iden-
ticado una signicativa correlación de la fracción
lábil de glomalina con la estabilidad estructural de
los agregados (Tabla 4).
Lozano et al. (2016), en un muestreo en dos loca-
lidades (Gorga y Gata) del SE-España, inmediata-
mente tras un incendio forestal de pino carrasco,
tampoco identican variaciones en el contenido
en glomalina lábil de los suelos, lo que atribuyen a
una baja severidad de la quema; los mismos auto-
res muestran como la glomalina lábil varía estacio-
nalmente en suelos de pinares no quemados.
Shari et al. (2018), en bosquetes de Quercus recién
quemados, observan como, a pesar de no variar
el contenido en carbono orgánico del suelo, las
fracciones de glomalina lábil y residual aumentan
con la severidad del incendio. También observan
un aumento de la relación glomalina/carbono or-
gánico, al igual que en este trabajo (con valores
en suelos control inferiores a 0,7 que aumentan a
Tabla 2 - Características de las quemas de matorral
Localidad Asín de Broto
Cubierta vegetal (%) 95
Biomasa (t/ha) 34,1
Quema 0 años (Q0) Marzo 2021
Quema 5 años (Q5) Noviembre 2016
Quema 9 años (Q9) Enero 2012
Tabla 3 - Fracciones de glomalina tras la quema prescrita de
matorral
Glomalina
Tsuelo
(cm)
Total
(mg g-1)
Residual
(mg g-1)
Lábil
(mg g-1)
L/Total
ratio
0-1 3,6ab 2,2a 1,4c 0,39c
C 1-2 3,6ab 2,5ab 1,1abc 0,31bc
2-3 2,9a 2,3a 0,6ab 0,23ab
0-1 7,8d 6,2e 1,6c 0,20ab
Q0 1-2 6,9cd 5,6de 1,3bc 0,19ab
2-3 5,6bc 4,5cde 1,2abc 0,21ab
0-1 5,0abc 4,3bcde 0,7ab 0,14a
Q5 1-2 4,5ab 3,9abcd 0,6a 0,14a
2-3 4,2ab 3,5abc 0,6ab 0,16ab
0-1 4,6ab 3,9abcd 0,65ab 0,14a
Q9 1-2 4,3ab 3,7abcd 0,55a 0,13a
2-3 3,9ab 3,4abc 0,55a 0,14a
F-valor 10,93 9,41 7,32 6,14
P-valor <0,001 <0,001 <0,001 <0,001
Letras distintas dentro de cada columna indican diferencias signicativas
(p<0,05) según el test de Tukey
Tabla 4 - Coecientes de correlación (r) entre las fracciones
de glomalina y diferentes propiedades edácas.
n=48
GR GL Corg CO2Cmic EE
GT 0,96** 0,53** 0,55** 0,44** 0,41** 0,64**
GR 0,28 0,40** 0,39** 0,30* 0,60**
GL 0,73** 0,35* 0,54** 0,38*
GT (glomalina total), GR (glomalina residual), GL (glomalina lábil),
Corg (carbono orgánico), CO2 (respiración basal), Cmic (carbono
microbiano), EE (estabilidad estructural). * p<0,05; **p<0,01
480 Revista de Ciências Agrárias
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2022,
45(4): 477-481
1,0 con la quema). Aunque los cambios a largo
plazo podrían relacionarse una menor actividad
degradativa de la glomalina por parte de los mi-
croorganismos o un aumento de la actividad fún-
gica con la sucesión vegetal, el brusco cambio en la
fracción residual post-quema solo parece explica-
ble por la liberación de glomalina ocluida en agre-
gados fragmentados tras el paso del fuego (Lozano
et al., 2016). Sin embargo, las quemas prescritas no
se caracterizan precisamente por generar un in-
tenso shock térmico, por lo que deben investigarse
otras causas para explicar las citadas variaciones.
Una posibilidad es que con la extracción de la glo-
malina se incluyan otros compuestos, incluso no
proteicos (Gillespie et al., 2011).
CONCLUSIONES
Se ha observado un aumento signicativo de las
fracciones de glomalina total y residual inmediata-
mente después de la quema prescrita de matorral;
la fracción lábil, no afectada justo tras la quema,
disminuye a los 5 y 9 años. Se postula la hipótesis
de que con el calentamiento hay una mayor extrac-
ción, aunque este tipo de quemas es de baja severi-
dad. Se requiere de una mayor investigación para
descartar que con la cuanticación de PSRG no se
cuantiquen también otros compuestos.
AGRADECIMIENTOS
A. Alfaro-Leranoz está contratado como personal
investigador predoctoral en formación por el De-
partamento de Ciencia, Universidad y Sociedad
del Conocimiento del Gobierno de Aragón. Por
su lado, la estancia del Dr. M. Emran en la EPS
de Huesca ha sido nanciada por el Ministerio de
Educación Superior e Investigación Cientíca de
Egipto. La toma de muestras no habría sido posible
sin la colaboración del Equipo de Prevención Inte-
gral de Incendios Forestales (EPRIF) de Aragón y
de los ganaderos locales.
481
Alfaro-Leranoz et al., Evolución de la glomalina tras la quema prescrita
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It has been speculated that arbuscular mycorrhizal fungi (AMF) produce extracellular compounds and that these are involved in soil stabilization. An unusual and abundant protein was found on hyphae of AMF, and it was hypothesized that the hyphal protein could be found in soil. The purpose of this work was to test extraction conditions for soil protein on a range of soils collected from undisturbed sites, to compare protein from hyphae with protein from soils, and to examine the relationship between soil protein and percent of total dry weight of soil made up of ca. 1 to 2-mm water stable aggregates. The A-horizons of 12 acidic soils collected from the midAtlantic area of the United States were used to develop a protocol for protein extraction. Protein was measured by the Bradford assay, with bovine serum albumin as the standard. Also, an indirect enzyme-linked immunosorbent assay (ELISA), using the monoclonal antibody that had been used to detect the protein on hyphae of AMF, was performed on 0.5 μg of protein from soil or hyphae. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of denatured protein was used to compare soil protein with hyphal protein. The extraction conditions that gave the best results for total protein were 0.25 g soil in 2 mL of 50 mM citrate, pH 8.0, for 90 min at 121°C yielding a range of 4.4 to 14.4 mg protein/g of soil for these soils. Highest ELISA values were obtained by using 20 mM citrate, pH 7.0, for 30 min at 121°C, but less protein was extracted under these conditions. Crude extracts of protein from hyphae and soil showed the same banding patterns and density of bands on SDS-PAGE gels. ELISA values for soils were between 60 and 107% of hyphal ELISA values. Total protein concentration was correlated linearly with organic carbon in soil (r2 = 0.85, P ≤ 0.001). The percent dry weight of soil composed of water-stable aggregates was correlated positively (P ≤ 0.05) with silt and ELISA values and correlated negatively with sand. The possible contribution of this protein to soil stabilization is discussed.
Article
Glomalin is reportedly a stable and persistent protein produced in copious quantities by mycorrhizal fungi and may be an important pool of organic N in soil. Glomalin-related soil protein (GRSP), however, is only operationally defined by its extraction method, and has been only poorly characterized at best. The goal of this study was to characterize the molecular structures within GRSP. Synchrotron-based X-ray absorption near-edge structure (XANES) spectroscopy and pyrolysis field-ionization mass spectrometry (Py-FIMS) revealed that GRSP contains a consortium of proteins along with many impurities. Employing proteomic techniques, we found that glomalin itself may be a thioredoxin-containing chaperone; however, no homologies with proteins or DNA of mycorrhizal origin were detected. Proteomics techniques further revealed that this fraction contains large amounts of soil-related heat-stable proteins and proteins of non-mycorrhizal origin. Results of this research show that the current extraction procedure that defines GRSP yields a mixture of compounds and thereby overestimates glomalin stocks when quantified using the Bradford assay. The chemical nature of glomalin has yet to be conclusively determined; it is unlikely that the chemical structure of glomalin can be elucidated from the mixture extracted as GRSP. Instead, an investigation into the specific biochemistry of immunoreactive assays currently used to define GRSP, followed by proteomic characterization of monoxenic mycorrhizal cultures may be required to advance our understanding of the chemical nature and agronomic significance of GRSP in soils.
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Arbuscular myc-orrhizae are important factors of soil quality through their effects on host plant physiology, soil ecological interactions, and their contributions to maintaining soil structure. The symbiosis is faced with numerous challenges in agroecosystems; in order to inform sustainable management strategies it is hence a high priority to work towards mechanistic understanding of arbuscular mycorrhizae contributions to soil quality. This review focuses on glomalin-related soil protein (GRSP), operationally defined soil C pools that have been linked to arbuscular mycorrhizal fungi (AMF). In discussing this protein pool, we propose a new terminology used to describe fractions of soil proteins and glomalin. GRSP concentrations in soil are positively correlated with aggregate water stability. GRSP has relatively slow turnover in soil, contributing to lasting effects on aggregation. Controls on production of GRSP at the phenomenological and mechanistic level are evaluated. While there are significant gaps in our knowledge about GRSP and glomalin (particularly at the biochemical level), it is concluded that research on GRSP holds great promise for furthering our knowledge of soil structure and quality, for informing suitable management, and as a foundation for novel biotechnological applications in agriculture and beyond.
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Glomalin, a substance produced by arbuscular mycorrhizal fungi, is reported to play a role in soil aggregation, but this role has been questioned in soils rich in calcium carbonate. We studied the relationship between aggregation stability and glomalin in a Haplic Calcisol comparing abandoned and active cultivation of olive groves. Abandonment was associated with increases in soil organic carbon, the percentage of water stable aggregates (WSA1–2mm), and easily extractable and total Bradford-reactive soil protein. WSA1–2mm was strongly positively correlated with both easily extractable and total Bradford-reactive soil protein. While easily extractable Bradford-reactive soil protein measured in both stable and unstable aggregates did not show any significant differences, Bradford-reactive soil protein was twice as high in stable than in unstable aggregates under both tillage and abandonment. Our results suggest that Bradford-reactive soil protein influences aggregate stability, even in soils with low organic matter and high calcium carbonate contents. However, more research is needed to elucidate the role of easily extractable Bradford-reactive soil protein in soil aggregation.