Agriculture, Ecosystems and Environment 185 (2014) 245–252
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Agriculture, Ecosystems and Environment
j ourna l h om epage: www.elsev ier .com/ locate /agee
iversity of arbuscular mycorrhizal fungi in Atlantic forest areasnder different land uses
amilla Maciel Rabelo Pereiraa,∗, Danielle Karla Alves da Silvaa,raeska Carenna de Almeida Ferreiraa, Bruno Tomio Gotob, Leonor Costa Maiaa
Programa de Pós-Graduac ão em Biologia de Fungos, Departamento de Micologia, Universidade Federal de Pernambuco, Avenida da Engenharia, s/n,ecife, PE 50740-600, BrazilDepartamento de Botânica Ecologia e Zoologia, Universidade Federal do Rio Grande do Norte, Campus Universitário, Natal, RN 59072-970, Brazil
r t i c l e i n f o
rticle history:eceived 17 July 2013eceived in revised form0 December 2013ccepted 3 January 2014vailable online 29 January 2014
eywords:lomeromycotacological indicesndicator species
a b s t r a c t
Agricultural land use compromises the viability of Atlantic forest remnants and may permanently alterthe structure of the biological soil community. Soil micro-organisms such as arbuscular mycorrhizal fungi(AMF) which participate in symbiotic associations with plant roots are of particular importance. In orderto assess the impact of cultivation we measured the diversity of AMF in six areas in Goiana, PE, Brazil:a sapodilla plantation, a rubber tree plantation, a mahogany plantation, a eucalyptus plantation, a croprotation area currently being used to cultivate cassava, and an area of Atlantic forest. A total of 96 samplesof rhizospheric soil were collected in the wet (June 2011) and dry (March 2012) seasons. Glomerosporeswere extracted from the soil, counted and used for AMF species identification. A total of 50 speciesbelonging to 15 genera were recorded. Acaulospora spp. and Glomus spp. predominated, accounting for52% of total species. The low value found in non-metric multidimensional scaling (NMS) multivariate
gricultural practices analyses (33.2%) indicated that AMF community composition was more affected by different land usesthan by physical and chemical characteristics of the soil. Diversity, evenness and richness indices werehigher for the environment under greater stress (crop rotation), indicating that mycorrhizal symbiosiscould be a strategy by which fungi and plants overcome biotic and abiotic stresses that occur in thesoil. Diversity, evenness and richness indices tended to be lower in communities established in climaxenvironments, such as in the Atlantic forest, rather than in the ones established in cultivation areas.
. Introduction
The Brazilian Atlantic forest is one of 25 world biodiversityotspots (Myers et al., 2000) and contains more species’ diversityhan most Amazonian forest formations (Morellato and Haddad,000). The Atlantic forest extends from Rio Grande do Norte toio Grande do Sul and encompasses a variety of different land
ormations, landscapes and climates (Silva and Casteleti, 2003).t is considered one of the two most threatened biomes on Earth.nly about 7% of the original vegetation remains today and what
s left has a highly fragmented distribution along the coast (SOS
ata Atlântica, 2013). Two processes that have led to massive
estruction of the forest, particularly in the northeast of Brazilre the cultivation of extensive areas and the process of intense
∗ Corresponding author at: Departamento de Micologia, Universidade Federal deernambuco, Avenida da Engenharia, s/n, Recife, PE 50740-600, Brazil.el.: +55 81 2126 8865; fax: +55 81 2126 8482.
urbanization, resulting in very small forest fragments widelyspaced one from another (Tabarelli et al., 2005; Wright andMuller-Landau, 2006; Ribeiro et al., 2009).
Compared to plants and animals relatively little is known aboutmicro-organisms diversity in Atlantic forest soils and the functionalroles they play in this biome. Among these organisms, arbuscularmycorrhizal fungi (AMF, Glomeromycota) form a mutualistic sym-biosis (Bonfante and Genre, 2010) in which the plant provides thefungus with energy for growth and maintenance through its photo-synthetic products, while the fungus provides water and nutrientssuch as phosphorus to the plant (Smith and Read, 2008).
The conversion of natural vegetation into agricultural fieldstriggers serious damage to the soil, such as negative influenceson the energy and biogeochemical cycles and changes in particleaggregation, as well as exposing it to insolation, erosion and nutri-ent leaching (Islam and Weil, 2000). These changes in vegetationcover have a global impact on biodiversity, soil degradation, and
the ability of biological systems to support human needs (Lambinet al., 2003).
These new land uses can also alter the structure of mycorrhizalcommunities in the soil, affecting their functions and hence
cosystem sustainability. In this regard AMF may be used as bio-ogical soil quality indicators (Oehl et al., 2004, 2010; Lamb et al.,005). Due to the agricultural and ecological importance of AMF,axonomic studies on the group have been intensified (Caruso et al.,012; van der Heijden et al., 1996; Oehl et al., 2011; Redecker et al.,013). In this context, the study of morphological and functionaliversity of AMF in Atlantic forest areas can contribute to increasednowledge on the distribution of these fungi, as well as provideseful information about their role in edaphic dynamics.
Assessing the impact of land use changes on AMF communi-ies is important for their management and for understanding theffects caused by this human action on the environment in order toenerate effective options for creating recovery strategies and/oriodiversity conservation. AMF are a key functional group in ter-estrial ecosystems but more studies are needed to qualitativelyvaluate their performance in the environment and understand theffects of conversion of natural areas to cultivation.
This study was designed to determine AMF diversity in Atlanticorest areas under different land uses and specifically to testhether agricultural practices negatively affect these micro-
rganisms and to see whether different systems of land useompromise AMF species diversity, evenness and richness.
. Material and methods
.1. Study areas
The study was conducted at the Itapirema Experimental Station,gronomic Institute of Pernambuco – -IPA, located in the cityf Goiana, Pernambuco, Brazil (07◦38′20′′S, 034◦57′10′′W, altitude3 m). The climate in the area is Ams′ (Köppen) type-rainy tropi-al monsoon with dry summer, with average annual precipitationnd temperature of 24 ◦C and 2000 mm, respectively. The soilype is Ultisol. Six areas under different soil use were selectedTable 1): sapodilla plantation (SA), rubber tree plantation (RT),
ahogany plantation (MA), eucalyptus plantation (EU), a crop rota-ion area currently producing cassava (CA), and an Atlantic forestrea (AF). These areas were divided in three classes of intensityf use according to the ongoing pressure factors such as planttilization, proximity of roads, soil disturbance and frequency ofertilization (Table 1).
.2. Sampling
Rhizosphere soil samples (0–20 cm deep) were collected in June011 and March 2012. In each of the five areas under different vege-ation cover (SA, RT, MA, EU, CA) and also in the reference area (AF),
plot of 1000 m2 and eight composite samples (five sub-samples atquidistant points) were collected around the host plants, packed inlastic bags and transported to the Mycorrhizae Laboratory/UFPE atmbient temperature. The soil collected was divided into portionsntended for: mounting trap cultures, AMF community assessment,nd physicochemical soil characterization. This last analysis waserformed at the Federal Rural University of Pernambuco, at theugarcane and Sugar Experimental Station of Carpina according toMBRAPA (1997). The results indicated that the soils had low levelsf phosphorus and organic matter (Table 2).
.3. Glomerospore analysis
Glomerospores were extracted from 50 g of field soil fromach sample by wet sieving followed by sucrose centrifugation
Gerdemann and Nicolson, 1963; Jenkins, 1964), and quantifiedsing a stereomicroscope (40×). In order to assist taxonomic anal-sis, trap cultures were set with soil collected from the field,n 2 L plastic pots with millet (Panicum miliaceum L.), corn (Zea
and Environment 185 (2014) 245–252
mays L.) and sunflower (Helianthus annuus L.) as hosts. The trapcultures were maintained in a greenhouse for three multiplica-tion cycles (four months each). At the end of each cycle, theplants were allowed to dry and cut off and aliquots of 50 g ofsoil were collected; after that, the soil was reseeded for the sub-sequent cycle. The aliquots of the soil collected at the end ofeach cycle were used for glomerospore extraction and subsequentAMF identification. For the taxonomic study, after counting, theglomerospores were mounted on microscope slides with PVLG(polyvinyl alcohol and lactoglycerol) and PVLG + Melzer’s reagent(1:1, v/v). Species identification was performed with the aid of adefined bibliography (Schenck and Perez, 1990), publications withdescriptions of new species and by consulting the international cul-ture collection of arbuscular mycorrhizal fungi database-INVAM(http://invam.caf.wvu.edu) and the on-line AMF collection ofthe Department of Plant Pathology, University of Agriculture inSzczecin, Poland (http://www.agro.ar.szczecin.pl/∼jblaszkowski/).In this work we adopted the classification proposed by Oehl et al.(2011), including recently described new taxa (e.g. Goto et al., 2012;Błaszkowski and Chwat, 2013).
2.4. AMF diversity analysis
AMF communities from field samples were evaluated bothquantitatively and qualitatively from population data (occurrenceand distribution frequency) and had their structure analyzed usingecological indices to measure species richness and diversity. Thefrequency of occurrence (FO) of the species was estimated accord-ing to the equation: Fi = Ji/k, where Fi = occurrence frequency ofspecies i, Ji = number of samples in which species i occurred andk = total number of soil samples. The species were classified asdominant (FO > 0.50), very common (FO between 0.31 and 0.50),common (FO between 0.10 and 0.30) and rare (FO < 0.10) (Zhanget al., 2004). The species were also classified as generalists (presentin all six areas), intermediate (present in two to five areas) or exclu-sive (present in one area) (Stürmer and Siqueira, 2011). Speciesrichness was measured as the ratio between the number of speciesobserved and the sample size, and the estimated number of specieswas calculated using the jackknife first-order index (Jackknife1).For the calculation of diversity in the study areas, the Shan-non index was used on a logarithmic base: H′ = �pi ln pi, wherepi = glomerospore number of each species/total glomerospores. Thespecies evenness was calculated using the Pielou evenness (J′)index, where: R = H′/log S, where H′ = value obtained using Shan-non and S = total number of AMF species present in the sample. TheSimpson dominance index (C) was calculated according to the for-mula C = �(ni(ni − 1)/N(N − 1) where ni = the abundance of species iand N = total abundance. Similarity between AMF communities wasassessed with the Sørensen index (Brower et al., 1990).
2.5. Statistical analyses
The data from soil physicochemical attributes were submittedto ANOVA and the means compared by the LSD test (p < 0.05) usingthe STATISTICA program (Statsoft, 1997).
To assess the impact of changes in land use on the AMF commu-nity, the NMS multivariate ordination method (non-metric multi-dimensional scaling) with Sørensen distance was used to explorethe relationship between soil properties and AMF distribution. Theanalysis of indicator species was randomized using the Monte Carlotest to determine which AMF species were sensitive to land use. Theindication value (VI) and the significance (p) value are products of
the relative abundance and occurrence frequency of species in eacharea (Dufrêne and Legendre, 1997). The values adopted in this workto consider a species as an indicator were VI > 50 and p < 0.001. NMSanalysis and the identification of indicator species were calculated
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Table 1Characterization of collection areas.
Area Character Rise time Plant species Fertilization Liming Intensity of use
Atlantic forest (AF) About 200 ha of remnant Atlanticforest. Dense ombrophylous forestvegetation
Unknown 120 tree species No No Lowa
Sapodilla plantation (SA) 2 ha abandoned area, with thepresence of grasses
50 years Manilkara zapota Forsberg NPK Limestone Mediumb
Rubber tree plantation (RT) 1 ha area preceded by pasture 32 years Hevea brasiliensis Muell. Arg., NPK Limestone MediumMahoganyplantation (MA) 2 ha area preceded by Atlantic forest 9 years Swietenia macrophylla King NPK Limestone LowEucalyptus plantation (EU) 2 ha area preceded by Atlantic forest 8 years Eucalyptus spp. NPK Limestone LowCrop rotation with cassava (CA) 2 ha area preceded by sugarcane
= 90 kg ha of ammonium sulphate, P = 90 kg ha Single superphosphate, K = 60 ka Low use = far from the roads, plants not used or managed.b Medium = near the roads, plants managed and rubber tree used for extraction oc High = annual tillage
sing PC-ORD version 5.0 software (McCune and Mefford, 2006)nd the species accumulation curve, cluster analysis and ecologi-al indices were calculated using Primer 6.0 software (Clarke andorley, 2006). The comparison of diversity indices was performedsing the PAST program, version 2.17 (Hammer et al., 2001).
. Results
.1. AMF community composition
Including the two collection periods and the six areas studied, 50 taxa weredentified (Table 3) belonging to 17 genera and 12 Glomeromycota familiesAcaulosporaceae, Archaeosporaceae, Ambisporaceae, Dentiscutataceae, Diversis-oraceae, Entrophosporaceae, Gigasporaceae, Glomeraceae, Intraornatosporaceae,acisporaceae, Racocetraceae, Scutellosporaceae). Of the total taxa, 44 (88%) weredentified at the specific level and six (12%) only at the generic level (Table 3).
Thirty-four species were recorded during the rainy season and 40 in the dry,nd the similarity of the AMF community between periods was 67.5%. The largestumber of identified species belonged to the genera Acaulospora and Glomus with3 and 12 species, respectively, followed by Racocetra with five and Gigaspora withour species. Ambispora, Cetraspora, and Fuscutata were represented by two speciesach, while Archaeospora, Claroideoglomus, Corymbiglomus, Dentiscutata, Funelli-ormis, Orbispora, Pacispora, Paradentiscutata, Scutellospora and Simiglomus wereepresented by only one species. The representativeness of genera corresponded to6% of Acaulospora, 24% of Glomus, 10% of Racocetra, 8% of Gigaspora, 4% of Ambispora,nd Cetraspora, and Fuscutata and 2% of other genera.
Four species (8%) found in all areas were classified as generalists: Acaulosporaoveata, A. scrobiculata, Gigaspora decipiens and Glomus macrocarpum. Twenty-seven
pecies were considered intermediate (54%) and 19 exclusive (38%). The highestumber of exclusive species occurred in the crop rotation area with cassava10 species) followed by the area of Atlantic forest (5 species) (Table 3). Thereas subjected to some type of agricultural practice had a higher number oflomerospores, with the exception of rubber tree plantations (Table 4).
able 2hysicochemical characterization of soil in areas of Atlantic forest (AF) and sapodilla planlantations (EU) and crop rotation with cassava (CA), in Itapirema Experimental Station –
Physicochemical characterization of soil AF SA
pH (H2O) 5.18 d 5.31cd
P (mg/dm3) 3.38 d 6.00 c
Fe (mg/dm3) 130.90 a 113.70 a
Cu (mg/dm3) 0.44 cd 0.63cd
Zn (mg/dm3) 2.10 c 3.35 bc
Mn (mg/dm3) 1.48 d 1.84 d
K (cmolc/dm3) 0.06 b 0.06 b
Na (cmolc/dm3) 0.05 a 0.04 a
Al (cmolc/dm3) 0.20 a 0.15 ab
Ca (cmolc/dm3) 1.00 c 1.31 bc
Mg (cmolc/dm3) 0.30 cd 0.36 bcd
H (cmolc/dm3) 2.06 b 2.59 a
S.B. (cmolc/dm3) 1.40 b 1.77 b
CTC (cmolc/dm3) 3.66 b 4.52 b
V (%) 38.30 b 37.70 b
C (%) 0.90 bc 1.05 ab
O.M. 1.56 bc 1.72 ab
= active acidity; SB = sum of bases; CTC = cation exchange capacity; V = base saturatiotatistically differ according to the Tukey test (5%).
of potassium chloride (added at planting seedlings).
.
Only three species had consistent differences regarding their presence in thetwo seasons (Table 3). Spores of Acaulospora foveata and Ambispora appendicula weremore abundant and frequent during the rainy season, while Acaulospora morrowiaewas very common during the dry season. Regarding the occurrence frequency,species of Acaulospora, Glomus, Gigaspora and Racocetra were dominant in all cul-tivated areas. Most species were found in less than 30% of the samples, presentingunusual or common forms.
Except for Glomus macrocarpum, no sporocarpic species (Glomus brohultii, G.coremioides, G. glomerulatum, G. sinuosum, G. taiwanense and G. sp. 2) occured inthe area with crop rotation (Table 4). Five species were considered indicators ofone of the areas: Cetraspora gilmorei (50.0) and Orbispora Pernambucana (62.5) wereindicators of the Atlantic forest, Gigaspora sp. 1 (68.3) and Racocetra tropicana (56.2)were indicators of the crop rotation area with cassava and Glomus sp. 2 (75) of themahogany plantation (Table 3).
3.2. Diversity
The area under crop rotation with cassava (CA) had the largest number of iden-tified species, with 30 taxa (Table 3), followed by the sapodilla (24), mahogany (19),Atlantic forest (17), rubber (16) and eucalyptus planting areas (16). According to thefirst-order jackknife richness estimator, the expected number of species was 42 inthe crop rotation with cassava, 32 in sapodilla, 28 in mahogany, 25 in the Atlanticforest, 21 in rubber and 24 in eucalyptus plantations (Fig. 1). The sampling effort ofthe present study was sufficient to recover 70–77% of the AMF species present inthe areas.
The Shannon diversity index was higher in the crop rotation area than inthe Atlantic forest (p < 0.05), while the Margalef’s diversity index separatedthe crop rotation area from all other areas. The Simpson’s dominance indexdiffered statistically among areas and was higher in the Atlantic forest, while the
evenness presented the lowest values in this area (AF) (Table 4). The non-metricmultidimensional scaling (NMS) analysis showed little relationship between soilphysicochemical properties and AMF community composition, since the proportionof the explained variance was low in both axes (Fig. 2). The axes of NMS analysisexplained only about 30% of the data variation, with the first axis explaining 16.8%
5.56 bcd 5.90 ab 6.05 a 5.65 abc21.10 a 8.50 c 8.93 c 12.9 b
104.60 ab 75.90 b 132.10 a 113.78 a4.09 a 0.22 cd 0.01 d 2.69 bc3.68 bc 4.61 ab 3.70 bc 6.07 a6.23 ab 7.49 a 3.36 c 3.99 bc0.14 a 0.06 b 0.05 b 0.04 b0.05 a 0.04 a 0.05 a 0.03 b0.04 bc 0.00 c 0.00 c 0.08 bc1.14 bc 2.31 a 2.25 a 1.55 b0.30 cd 0.44 b 0.72 a 0.30 d2.13 ab 2.18 ab 1.97 b 1.56 c1.62 b 2.85 a 3.06 a 1.93 b3.78 b 5.02 a 5.03 a 3.56 b
43.40 b 56.60 a 60.50 a 53.60 a0.90 bc 1.08 a 1.02 ab 0.76 c1.55 bc 1.86 a 1.77 ab 1.31 c
n; O.M. = organic matter. Means followed by the same letter, in the row, do not
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Fig. 1. Species accumulation curve (Sobs) and richness estimation by the first order jackknife index (jackknife 1), in the Atlantic forest areas (A) and sapodilla (B) rubber (C),mahogany (D), eucalyptus (E) plantations and crop rotation with cassava (F).
C.M.R. Pereira et al. / Agriculture, Ecosystems and Environment 185 (2014) 245–252 249
Table 3Frequency of occurrence and indicator species index of AMF species in Atlantic forest areas (AF) and sapodilla (SA), rubber (RT), mahogany (MA), eucalyptus (EU) plantationsand crop rotation with cassava (CA), in the Experimental Station of Itapirema - IPA, Brazilian northeastern.
Frequency of Occurrence Indicator Species
AMF Species AF SA RT MA EU CA Area Indicator value P
AcaulosporaceaeAcaulospora elegans Trappe & Gerd. – R – R – – MA 9.1 1.0A. excavata Ingleby & C. Walker – R D C R D RT 35.6 0.01A. endographis B.T. Goto – – – – – R CA 12.5 1.0A. foveata Trappe & Janos R MC MC R R MC SA 31.2 0.01A. longula Spain & N.C. Schenck – – – – – R CA 12.5 1.0A. mellea Spain & N.C. Schenck R MC MC – R MC RT 28.3 0.1A. morrowiae Spain & N.C. Schenck – C C – C C EU 22.7 0.1A. scrobiculata Trappe R C MC MC C D CA 32.2 0.02A. spinosa Walker & Trappe – R – – – R CA 12.5 1.0A. tuberculata Janos & Trappe – R R – – C CA 21.4 0.14Acaulospora sp.1 – C D – – – SA 21.7 0.13Acaulospora sp.2 – C C – – R RT 21.9 0.12Acaulospora sp.3 – – – – – R CA 12.5 1.0ArchaeosporaceaeArchaeospora trappei Ames & Linderman R – – – – – SA 8.2 1.0AmbisporaceaeAmbispora apendicula Spain, Oehl & Sieverd – C C R C MC CA 40.9 0.01A. gerdermanii (S.L. Rose, B.A. Daniels & Trappe) J.B. Morton & D. Redecker – R – R – EU 12.5 1.0DiversisporaceaeCorymbiglomus tortuosum (N.C. Schenck et G.S. Sm.) Błaszk. et Chwat – – – – – R CA 42.3 0.006DentiscutataceaeDentiscutata cerradensis Spain & J. Miranda – – – C R – MA 9.4 1.0Fuscutata aurea Oehl, C.M. Mello & G.A. Silva – C – – – C SA 20.7 0.1F. heterogama Oehl, F.A. Souza, L.C. Maia & Sieverd. – – – – – C CA 23.7 0.07EntrophosporaceaeClaroideoglomus etunicatum (W.N. Becker & Gerd.) C. Walker & A. Schüßler R – – – – – AF 12.5 1.0GigasporaceaeGigaspora decipiens Hall & Abbott – R – MC C C CA 33.3 0.04G. gigantea Gerd. & Trappe C D R C D MC CA 38.5 0.01G. margarita Becker & Hall – C – MC – C EU 27.6 0.07Gigaspora sp.1 – – – – – D CA 68.3 0.0003GlomeraceaeFunneliformis mosseae (T.H. Nicolson & Gerd.) C. Walker & A. Schüssler R – – R – – AF 12.5 1.0Glomus aggregatum N.C. Schenck & G.S.Sm. – – C – – – SA 36.7 0.01G. ambisporum G.S.Sm. & N.C. Schenck – – – – – R CA 12.5 1.0G. brohultii Herrera, Sieverding & Ferrer R – – – – – AF 12.5 1.0G. clarum T.H. Nicolson & N.C. Schenck – R – – – –G. coremioides (Berk. & Broome) D. Redecker & J.B. Morton MC – – – C – AF 23.1 0.1G. glomerulatum Sieverding C – C R – SA 34.4 0.01G. macrocarpum Tul. & C. Tul. MC D MC D D CG. sinuosum (Gerd. & Bakshi) Almeida & Schenck – – – C R – MA 21.4 0.1G. taiwanense (Wu & Chen) Almeida & Schenck C C – C – – AF 5.0 1.0G. trufemii B.T. Goto, G.A. Silva & Oehl – C R C R C RT 9.5 1.0Glomus sp.1 – R – C – – MA 9.4 1.0Glomus sp. 2 (Sporocarpic) – – – MC – – MA 75 0.0001Simiglomus hoi (S.M. Berch & Trappe) G.A. Silva, Oehl & Sieverd. – – – – – R CA 12.5 1.0IntraornatosporaceaeParadentiscutata maritima B.T. Goto, D.K. Silva, Oehl & G.A. Silva – – – – – C CA 37.5 0.01PacisporaceaePacispora boliviana Sieverd. & Oehl C R – – – R AF 10.0 0.5RacocetraceaeCetraspora gilmorei (Trappe & Gerd.) Oehl, F.A. Souza & Sieverd. C – – – – – AF 50.0 0.001C. pellucida T.H. Nicolson & N.C. Schenck) Oehl, F.A. Souza & Sieverd. – – R – R – RSE 12.5 1.0Racocetra alborosea (Ferrer & R.A. Herrera) Oehl, F.A. Souza & Sieverd. – R – – – –R. coralloidea (Trappe, Gerd. & I. Ho) Oehl, F.A. Souza & Sieverd. – – – C – R MA 30.5 0.03R. fulgida (Koske & C. Walker) Oehl, F.A. Souza & Sieverd. C C – – – R RT 10.6 0.6R. tropicana Oehl, B.T.Goto & G.A.Silva – C – MC C D CA 56.2 0.0009R. weresubiae (Koske & C. Walker) Oehl, F.A. Souza & Sieverd – – – – – R CA 12.5 1.0ScutellosporaceaeOrbispora pernambucana Oehl, D.K. Silva, N. Freitas & L.C. Maia MC – – – – – AF 62.5 0.0002Scutellospora aurigloba C. Walker & F.E. Sanders C – – – – R AF 16.7 0.4
Species richness 17 24 16 19 16 30
D
oCvw
= dominant; MC = most common; C = common and R = rare.
f the variation correlated negatively with Fe, Al and positively with Zn, Mn, pH,a, Mg, SB, CTC, V, C, MO, total sand and silt while axis 2 explained 16.4% of theariation and was negatively correlated with Cu, Mn, P, K and positively correlatedith Ca, Mg and CTC (Table 5).
4. Discussion
The six taxa identified only at the genus level probably are newto science, considering that the characteristics did not resemble any
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Table 4Number of glomerospores (in 50 g of soil), diversity index, evenness and dominance of AMF communities in Atlantic forest areas (AF) and sapodilla (SA), rubber (RT), mahogany(MA), eucalyptus (EU) plantations and crop rotation with cassava (CA), in the Itapirema Experimental Station-IPA, northeastern Brazil.
Areas AF SA RT MA EU CA
Number of glomerospores* 87.0 ab 124.0 a 63.0 b 138.0 a 137.0 a 60.0 bMargalef’s index (d)** 3.102 bc 3.885 b 2.685 c 3.297 bc 3.317 bc 4.794 aShannon index (H′)** 1.832 b 2.357 a 2.349 a 2.462 a 2.181 ab 2.533 aPielou evenness (J′)** 0.660 c 0.752 b 0.867 a 0.836 a 0.787 b 0.752 bSimpson dominance (�)** 0.318 a 0.161 b 0.124 b 0.130 b 0.161 b 0.134 b
Means followed by the same letter, in the row, do not statistically differ according to tpermutation (0.1%).
Fig. 2. Non-metric multidimensional scaling analysis based on the communityof arbuscular mycorrhizal fungi correlated with soil physicochemical attributesieE
ose11
TC2
n
n areas of Atlantic forest (AF) and sapodilla (SA), rubber (RT), mahogany (MA),ucalyptus (EU) plantations and crop rotation with cassava (CA), in the Itapiremaxperimental Station – IPA, Brazilian northeastern.
ther known AMF species. The number of species recorded in thistudy (50) was sometimes superior to those found in other natural
cosystems and in different areas converted to crop fields. In Brazil,0 AMF species have been recorded in the Cerrado (Alvarenga et al.,999) and 61 in the Amazon region (Stürmer and Siqueira, 2011).
able 5orrelation coefficient of the variables analyzed (secondary matrix) with axes 1 and
of the NMS ordination.
Variables Correlation coefficient
NMS – -axis 1 NMS – -axis 2
Fe −0.340** 0.022 nsCu 0.012 ns −0.406**
Zn 0.399** −0.006 nsMn 0.440*** −0.243*
P 0.155 ns −0.556***
pH 0.699*** −0.004nsK 0.133 ns −0.465***
Na −0.150 ns −0.006 nsAl −0.707*** 0.051nsCa 0.679*** 0.266*
Mg 0.500*** 0.266*
H −0.068 ns 0.152 nsS.B. 0.686*** 0.232 nsCEC 0.533*** 0.303*
s: not significant at 5% probability.* Significant at 5% probability.
** Significant at 1% probability.*** Significant at 0.1% probability.
he *Tukey test (5%) and **by the randomization procedures of bootstrapping and
Worldwide, 39 AMF species have been reported in the temperateand dry tropical forests of Mexico (Gavito et al., 2008; González-Cortés et al., 2012), 37 in high plains in central Europe (Oehl et al.,2004) and 12 in rainforest in Kenya converted to agriculture (Jefwaet al., 2012).
Knowledge of AMF diversity in Atlantic forest areas in differ-ent states of conservation and land use is an important biologicalparameter for the assessment of environmental disturbances. Ofthe 250 AMF species described around the world (Oehl et al.,2011), 119 have been recorded to Brazil (de Souza et al., 2008).Of these, 78 were recorded in Atlantic forest areas (Zangaro andMoreira, 2010) and 79 in Brazilian agro-ecosystems (Carrenho et al.,2010). However, there are few studies investigating the impact ofthe conversion of natural vegetation to crop production. In thisregard, it is worth mentioning that the four managed forest areasdid not differ in terms of their AMF community structure, thoughthey did differ in terms of their community composition, demon-strating the importance of the plant host on the selection of AMFspecies.
Generally, species of Glomus and Acaulospora are more com-mon in both natural and managed environments (Aidar et al.,2004; Jefwa et al., 2012; Oehl et al., 2003). This is due to both thehigh number of described species in these genera as well as theirspread and adaptability (Daniell et al., 2001). Glomus (lato sensu)species mainly dominate in disturbed environments because theirhigh sporulation rate enables their colonization of these environ-ments, regardless of climatic conditions (Caproni et al., 2003).Acaulospora species are better established in soils with acidic pHranges (Sieverding, 1991) characteristic of tropical regions. How-ever, some Glomus species seem to be affected by the type of landuse, such as sporocarpic species (Tchabi et al., 2008), as thesespecies were missing in area with greater intensity of use (croprotation), due possibly to the prolonged dormancy of spores formedin sporocarps (Santos and Carrenho, 2011).
The occurrence frequency provides some indication of howadapted a species is to various environmental and soil conditions(Stürmer and Siqueira, 2008). Acaulospora foveata, A. scrobiculataand Glomus macrocarpum have been considered generalists inprevious studies in the Atlantic forest environment and in its sur-roundings (Silva et al., 2006; Trufem, 1990; Trufem et al., 1994).The dominance of these species in several different environmentsindicates high plasticity and high adaptation to different impactsof biotic or abiotic origin (Zangaro and Moreira, 2010).
The presence of different plant species, recurring preparationcycles and collection in the system with crop rotation (CA) main-tain the area in constant instability, inducing the AMF to frequentlyproduce propagules to ensure their survival (Carrenho et al., 2010).Abbott and Gazey (1994) have suggested that the intensity of soiluse modifies AMF species composition with diversity being smallerin cases of minimal disturbances. The low glomerospore density
and AMF species richness in the Atlantic forest area (AF) may bea consequence of its greater stability, less competition for fungalniches and prevalence of species with low sporulation that arenot pioneers (k strategist species). Greater glomerospore density
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C.M.R. Pereira et al. / Agriculture, Ecosy
nd species richness equal or superior in other areas indicate thatrolonged monoculture selects fungi of rapid growth and sporu-
ation, with high adaptability (r strategist species), with selectionccurring for survival and not for efficiency in the host (Moreirand Siqueira, 2002). At the same time, the Atlantic forest soil hasharacteristically low natural fertility (Siqueira and Saggin-Júnior,001), which causes high dependence of many plant species toycorrhizal association, as demonstrated, for example, in cassava,
ven with constant additions of chemical inputs.In the present work, the species accumulation curves did not
each the stabilization point (plateau), indicating that the samplingffort did not retrieve all the AMF species present in the areas. How-ver, the sampling effort was enough to assess between 70 and7% of the estimated species for the areas by the jackknife index 1.
n natural and managed areas in the Amazon region, these curvesere also applied, and in most areas the plateau was not reached
Stürmer and Siqueira, 2011). The AMF total richness is sometimesnderestimated because many species may be present only in theegetative form and species are identified primarily from the glom-rospores (Bartz et al., 2008). Thus, we suggest new samplings andhe use of trap cultures for a longer period, which would enable theermination of species not found in field samples.
Regardless of the physical and chemical characteristics of theoil, species diversity was higher in cultivated areas, mainly inhe crop rotation area currently being used for the cultivation ofassava. Crop rotation involves a short period of bare soil, a highurnover of plant species and more frequent addition of fertilizer.hese factors may have potentially contributed to the increase ofMF diversity (Hijri et al., 2006).
The calculation of the diversity index takes into considerationwo important attributes of the community: rare species and rela-ive abundance. The Shannon index used herein, is one of the mostidely used in studies of AMF communities (Jefwa et al., 2012; Silva
t al., 2012; Stürmer et al., 2013) because it gives greater valueo rare species and is considered ideal for studying the disruptionffects suffered by the environments since they are the first to feelhe impacts (Caproni et al., 2003).
Of the five species considered indicators, or in other words,he most characteristic of a particular area, two were from thetlantic forest. The replacement of the Atlantic forest by agri-ultural crops raised the AMF diversity, as also demonstrated bytürmer and Siqueira (2011) in the Amazon region. However,espite the Atlantic forest area (AF) having a lower diversity thanhe cultivated areas, this area had a high number of unique speciesArchaeospora trappei, Claroideoglomus etunicatum, Glomus brohul-ii, Cetraspora gilmorei and Orbispora pernambucana). Therefore, thisvidence supports the importance of conservation of natural areas,specially those threatened with extinction, such as the Atlanticorest, in order to maintain species found only in these environ-
ents.The weak relationship between the physicochemical properties
f soil and AMF community composition revealed by NMS multi-ariate analysis has also been observed in other areas of subtropicalnd temperate forests converted into cultivated areas (Alguacilt al., 2008; González-Cortés et al., 2012). The area that turnednto an agricultural field with greater intensity of use (CA) changedhe AMF community more than areas with use less intense. Whenonsidering all areas studied, the Atlantic forest shared only 40% ofMF species with the other areas.
The results of the NMS analysis are in agreement with thosebtained from the similarity calculation (data not shown), suggest-ng that land use is more important than the chemical–physical
roperties of the soil in determining the AMF community compo-ition. However, we found significant correlations between someoil variables analyzed and the ordination axes. The content of Fend Al observed in the Atlantic forest may have contributed to the
and Environment 185 (2014) 245–252 251
area separation on axis 1, while cultivated areas showed an increasein pH and levels of Mn, Ca, Mg, sand, silt and organic matter, possi-bly due to agricultural practices that were performed and fertilizerapplied. Regarding the ordination axis 2, the levels of Cu, Mn, P andK contributed to differentiate the cultivated areas with higher con-centrations of these elements (MA and EU areas), while CTC, Ca andMg separate the rubber planting from other areas on axis 2.
5. Conclusions
The AMF community was more influenced by the land use thanthe physicochemical properties of the soil. We confirmed that thediversity, evenness and richness index tended to be lower in com-munities established on vegetation climax environments than inagricultural crops. Despite the greater number of species in culti-vated areas, the conditions in these areas promoted the selectionof common AMF species, and led to the loss of species in areasconsidered hotspots of biodiversity.
Acknowledgments
The authors acknowledge: the Instituto Agronômico dePernambuco for logistical support; the Conselho Nacional deDesenvolvimento Científico e Tecnológico (CNPq) for Master andPhD scholarships and a research fellowship provided to C.M.R.Pereira, D.K.A. Silva and L.C. Maia, respectively, and financing ofprojects included in PROTAX, SISBIOTA, and INCT-Virtual Herbar-ium. The authors are indebted also to Inácio Monte Júnior andAngelo Santana for helping during soil collections; Everardo V.S.B.Sampaio for suggestions to improve the manuscript; Scott Healdand David Bousfield for English’s revision of the manuscript.
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