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Mycorrhizal fungi arbuscular 
in forage grasses cultivated 
in Cerrado soil
Leidiane dos Santos Lucas1,2, Aurelio Rubio Neto1, Jadson Belem de Moura2,5*, 
Rodrigo Fernandes de Souza2,3, Maria Eduarda Fernandes Santos2, 
Lorena Fernandes de Moura4, Elitania Gomes Xavier2, José Mateus dos Santos2,5, 
Ryan Nehring6 & Sandro Dutra e Silva3,5
The Cerrado is one of the most important regions for agricultural development in the world and is the 
main productive breadbasket of the Americas. One of the main agricultural activities in the region is 
high-tech livestock. Cerrado soils are predominantly low in fertility, and arbuscular mycorrhizal fungi 
play a fundamental role in plant nutrition in this biome. Understanding the behavior of mycorrhizal 
fungi in the soil under pasture is essential for the development of more efficient and sustainable 
management practices. Thus, this work aims to verify the activity of arbuscular mycorrhizal fungi 
in different species of forage grasses cultivated in cerrado soil. To measure mycorrhizal activity, 
soil spore density factors and mycorrhizal colonization rates in roots of 14 forage grass genotypes 
were investigated. No significant differences were identified in spore density values between the 
investigated genotypes. Panicum maximum cv and Mombasa showed the lowest values of mycorrhizal 
colonization, and the highest values were found in the roots of Brachiaria decumbens. Among the 
identified genera associated with the rhizosphere of the genotypes studied, Gigaspora, Scutelospora 
and Sclerocysts are less frequent, which indicates that the association with these fungal genera is less 
recurrent than with the others.
The Cerrado is the second largest Brazilian biome, extending over an area of 2,045,064  km2 and spanning eight 
states of Central Brazil: Minas Gerais, Goiás, Tocantins, Bahia, Maranhão, Mato Grosso, Mato Grosso do Sul, 
Piauí and Distrito  Federal1. It is divided by three of the largest hydrographic basins in South America, with 
regular rainfall indices that provide great biodiversity. After the Amazon, the Cerrado today is considered the 
last agricultural frontier of the  Americas2–4.
The Cerrado is considered to be a “biodiversity hotspot”, as it has one of the greatest levels of biodiversity 
on the planet. Such levels of biological diversity are achieved due to it being a transition biome that is in direct 
geographic contact with other important South American biomes, such as the Amazon, Caatinga, Atlantic 
Forest, Pantanal and Bolivian  Chacos2,5. From a natural history perspective, the Cerrado could be considered a 
biogeographic region that is more than 40 million years  old6–9. Such biogeographic continuity has resulted in a 
symbiosis between flora, fauna and  microorganisms3. Due to its privileged location, the Cerrado stands out as 
one of the most important agricultural frontiers in the  world3,4, with much of the area today consisting of large-
scale industrial agriculture and degraded  pastures10,11.
With the current productive paradigm, environmental sustainability is considered to be an important factor 
in determining the success of production systems. In agriculture, no-tillage systems are being promoted as a 
cultivation system that promotes soil and water  conservation12,13. For no-tillage systems to maintain levels of 
productivity, vegetation cover is an adequate factor. For conditions in the Cerrado, vegetation cover must have a 
low carbon–nitrogen ratio, which decreases the decomposition speed and increases the time in which the cover 
protects the soil from erosive  processes14. In this sense, grasses stand out as ideal plant cover for no-tillage sys-
tems in the Cerrado, in addition to being important forages for animal  grazing15. Natural cover systems based 
OPEN
1Graduate Studies in Agricultural Sciences / Agronomy, Instituto Federal Goiano, Rio Verde, Goiás, Brazil. 2Sedmo 
- Soil Research Group, Ecology and Dynamics of Organic Matter, Evangelical College of Goianésia, Goianesia, 
Goiás, Brazil. 3Graduate Studies in Natural Resources of the Cerrado, State University of Goiás, Anápolis, Goiás, 
Brazil. 4SEMPA Seeds Technology, Goiania, GO, Brazil. 5Graduate Studies in Social, Technological and Environment 
Science, Evangelical University of Goiás, Anápolis, Goiás, Brazil. 6Department of History and Philosophy of 
Science, University of Cambridge, Cambridge, UK. *email: jadsonbelem@gmail.com
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on symbiotic systems may also reduce the use of herbicides, which have been used extensively for no-till systems 
in the tropical climates of  Brazil16.
Generally, Cerrados are environments that naturally offer adverse abiotic conditions for plant growth and 
development. With low phosphorus levels and irregular rainfall, vegetation depends directly on the performance 
of mycorrhizal fungi to resist surviving such conditions, which is attributed to the association between fungi and 
plants as an important factor in building resilience to stressful  situations1,17–19.
The association of mycorrhizal fungi with vegetation started its evolution in tropical regions, and there are 
even species that are found only in these  regions20. Today, however, the presence of these fungi is reported in 
different regions of the planet, regardless of  climate19,21–25.
The average density of mycorrhizal fungal species in the soils of the Cerrado varies from 25 to 50 spores per 50 
 cm3 of soil on average. In the neighboring region of the Caatinga, there is a variation in the number of propagules 
of these fungi, probably due to differences in the plant community, and in relation to chemical composition and 
land use, with ranges containing high  phosphorus6,26.
For agricultural production, combined with the recovery of degraded areas, understanding the behavior of 
forage grass species with soil biology is essential for the development of more efficient practices for the manage-
ment of natural resources. Cerrados are environments that offer adverse abiotic conditions for plant growth and 
development, with low levels of phosphorus and a limited water regime, and the development of their vegetation 
depends directly on the action of soil microorganisms. Under these conditions, mycorrhizal fungi stand out as 
organisms that promote plant growth and contribute to plant resilience to stressful  situations1,17–19,27. Therefore, 
this work aims to verify the mycorrhizal population dynamics in forage grass species in Cerrado soils.
Materials and methods
The experiment was conducted at the Agrostological Field of the Ricardo Fontoura Experimental Station of the 
Cerrado, which is part of the Evangelical College of Goianésia, in the state of Goiás, Brazil. The climate is classi-
fied as a tropical season (AW) characterized by two well-defined seasons: dry and  rainy14. The density of spores 
and the mycorrhizal colonization rate of 14 varieties of forage grasses were evaluated (Table 1).
Samples of rhizospherical soil containing the treatment roots described in Table 1 were collected. Each sam-
ple taken to the laboratory was composed of 3 simple samples randomly collected from each plot. The design 
had a completely updated design with 6 replicates. Sampling was carried out at the end of the dry season in 
September 2020.
The analyses were carried out in the laboratory of agricultural microbiology of the Evangelical College of 
Goianésia. The spores of arbuscular mycorrhizal fungi (AMF) were extracted from 50  cm3 of rhizospherical 
soil by wet  sieving15 followed by centrifugation in water and a 50% sucrose solution. The spores were separated 
according to their phenotypic characteristics, such as color, size and shape, composing the different morphotypes 
under stereoscopic binocular magnifying  glass28.
To determine the percentage of colonization, the roots were clarified and ordered with 0.05% Trypan Blue 
in  lactoglycerol16 and the colonization was evaluated under under a stereoscopic microscope, following the 
quadrant intersection  technique17.
To identify the genera of AMF from morphological characteristics, the spores were separated according to 
their morphotypes and mounted on blades with pure polyvinyl-lactoglycerol (PVLG) and PVLG mixed with 
Melzer (1:1 v/v). To support the identification work, original articles from the descriptions of species were pro-
vided on the website of the "International Culture Collection of Arbuscular and Vesicular–Arbuscular Mycor-
rhizal Fungi"18,29.
Table 1.  Forage grasses installed at the Agrostological Field of the Ricardo Fontoura Experimental Station of 
the Cerrado, Evangelical College of Goianésia.
Forage Grasses
Urochloa decumbens
Brachiaria Ruziziensis
Brachiaria brizantha cv marandu
Brachiaria brizantha cv piatã
Brachiaria brizantha cv. Xaraes
Brachiaria brizantha cv. Paiaguas
Brachiaria brizantha cv. Ipyporan
Brachiaria brizantha cv. Humidicola
Megathyrsus maximus cv. Mombasa
Megathyrsus maximus cv. Kenya
Megathyrsus maximus cv. Zuri
Megathyrsus maximus cv. Aruana
Megathyrsus maximus cv. Tamani
Megathyrsus maximus cv. Massai
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The data were submitted to variance analysis by the  Assistat30 analyses of canonical correspondence were 
performed by the  Past20,31 software. Spore density variables, and the rate of mycorrhizal colonization was deter-
mined by a 5% Tukey test. The presence of identified genera was used as the parameter for multivariate analysis.
The conduct of the experiment followed the International guidelines of the IUCN Policy Statement on 
Research Involving Species at Risk of Extinction. The plant material studied, as a perennial species, is available 
for study and review at the agrostological field of the Evangelical College of Goianésia.
Results and discussion
For the determination of ecological interactions between AMF and forage grasses, the values of density of spores 
in the soil, mycorrhizal colonization rate in the root, and the presence of genera of AMF associated with the 
rhizosphere are used as parameters. No significant difference was verified between the analyzed varieties when 
investigating the density of spores in rhizospherical soil of forage grass varieties in Cerrado soils (Fig. 1a). P 
values for spore density were 0.2868 and for colonization were 0.1662.
The mycorrhizal colonization rate showed a significant difference (p < 0.05). Urochloa decumbens presented 
the highest mycorrhizal colonization rate (78%) compared to the others. The species Megathyrsus maximus cv. 
Zuri, Megathyrsus maximus cv. Aruana and Megathyrsus maximus cv. Mombasa presented the lowest values of 
mycorrhizal colonization in its roots, 33%, 34% and 27%, respectively (Fig. 1b).
The absence of a significant difference in spore density values is because the varieties were installed in the same 
area and where they are probably being colonized by the same fungal species, since they present low specificity. 
It is expected that there is no difference in the sporulation of fungi from the same area, since spore production 
103
109
125
111
140
152
137
91
104
92
120
117
101
113
0 20 40 60 80 100 120 140 160
Brachiaria decumbens
Brachiaria ruziziensis
Brizantha marandu
Brizantha piatã
Brachiaria brizantha cv. Xaraes
Brachiaria brizantha cv. Paiaguas
Brachiaria brizantha cv. Ipyporã
Brachiaria brizantha cv. Humidicola
Panicum maximum cv. Mombaça
Panicum maximum cv. Quenia
Panicum maximum cv. Zuri
Panicum maximum cv. Aruana
Panicum maximum cv. Tamani
Panicum maximum cv. Massai
No. of spores / 50 mL of soil
78% a
66% ab
45% ab
63% ab
65% ab
66% ab
45% ab
55% ab
27% b
43% ab
33% b
34% b
48% ab
45% ab
0% 10% 20% 30% 40% 50% 60% 70% 80% 90%
Brachiaria decumbens
Brachiaria ruziziensis
Brizantha marandu
Brizantha piatã
Brachiaria brizantha cv. Xaraes
Brachiaria brizantha cv. Paiaguas
Brachiaria brizantha cv. Ipyporã
Brachiaria brizantha cv. Humidicola
Panicum maximum cv. Mombaça
Panicum maximum cv. Quenia
Panicum maximum cv. Zuri
Panicum maximum cv. Aruana
Panicum maximum cv. Tamani
Panicum maximum cv. Massai
Mycorrhizal Colonization (%)
a
b
Figure 1.  Density of mycorrhizal fungi spores in rhizospherical soil (a) and rate of mycorrhizal colonization (b) 
of different forage grasses in Cerrado soils.
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is a response of the fungus and not of the host plant. The production of spores is a reflection of the fungus to 
environmental changes that, under stressful conditions, begin to produce spores as a resistance  structure21,22,32.
The samplings were carried out at the end of the dry season. In the cerrado, the climate is classified as tropical 
seasonal (AW) characterized by two well-defined seasons: a dry period and a rainy  period33. These climatic condi-
tions are considered stressful for most soil organisms due to the absence of rainfall for more than 4 consecutive 
months, which explains the high values of spore density values in the  soil12,23.
Forage plants do not present specificity for the colonization of mycorrhizal fungi and can be colonized by 
more than one species of  fungus24. However, some plant species have higher mycorrhizal colonization rates 
than others. Different species may present different values of colonization in the same environment, which is a 
reflection of the evolutionary adaptability of this symbiotic  association34–37.
Mycorrhizal colonization values indicate the intensity to which fungi have to associate with vegetation to 
assist with functions such as water and nutrient absorption (MOREIRA; SIQUEIRA, 2006). Because it is the 
same soil, the variation in mycorrhizal colonization values is explained by the physiological differences of plants 
and not fungi. This behavior can be observed when comparing forage plants, such as Megathyrsus maximus and 
Brachiaria brizanta, which presented similar colonization rates, regardless of  cultivar38–40.
Table 2.  Presence (1) and absence (0) of genera of arbuscular mycorrhizal fungi associated with the 
rhizosphere of different forage grasses in Cerrado soil.
Forage Acaulospora Claroideglomus Diversispora Scutellospora Sclerocystis Glomus Gigaspora
B. decumbens 1 1 1 1 0 1 1
B. Ruziziensis 1 1 1 0 0 1 1
B. Brizantha Marandu 1 1 1 1 0 1 1
B. Brizantha Piatan 1 1 1 0 1 1 1
B. brizantha cv. Xaraes 1 0 1 1 0 1 1
B. brizantha cv. Paiaguas 1 1 0 1 0 1 1
B. brizantha cv. Ipyporan 1 0 1 1 0 1 1
B. brizantha cv. Humidicola 1 0 1 0 0 1 1
P. maximum cv. Mombasa 1 0 0 0 0 1 1
P. maximum cv. Kenya 1 1 1 0 0 1 0
P. maximum cv. Zuri 1 1 0 1 0 1 0
P. maximum cv. Aruana 1 1 1 1 0 1 0
P. maximum cv. Tamani 1 1 1 1 0 1 1
P. maximum cv. Massai 1 1 1 0 0 1 0
Figure 2.  Canonical correspondence analysis of the associated genera found in rhizospheric soil of different 
forage grasses in cerrado soil.
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Table 2 shows the genera identified in the soil of the grasses investigated. The genera Acaulospora and Glomus 
were identified in all plants investigated, while the genus Slerocystis was identified to be associated only with 
B. brizantha piatã.
Canonical correspondence analysis aims to identify the proximity of the presence of mycorrhizal fungi identi-
fied with the grass species investigated (Fig. 2).
The genera of mycorrhizal fungi identified were commonly found in the rhizosphere of all grasses investigated, 
except for the genera Gigaspora, Scutelospora and Sclerocysts, which indicates that the association with these 
genera of fungi is less recurrent than with the other genera. The genera Glomus and Acaulospora are commonly 
found in Cerrado  soils6. When investigating the biodiversity of AMF in Cerrado soils, the same genera were 
found to be associated with  bamboo41,  sugarcane42,43, sorghum and  corn44.
The grasses present substantial mycorrhizal colonization volume for the root system of the grasses, which 
favors fungal colonization. The adaptability of this plant family to the natural conditions of the Cerrado also 
favors the exposure of the plant to the action of the fungus when subjected to situations of environmental stress, 
especially water.
Conclusion
The spore density values do not vary among the species of fodder studied. This parameter is independent of the 
plant, as it is a physiological response of the fungus. On the other hand, Urochloa decumbens presented higher 
values of mycorrhizal colonization. The genera of mycorrhizal fungi identified are commonly found in the 
rhizosphere of all grasses investigated, except for the genera Gigaspora, Scutelospora and Sclerocysts, and these 
genera of fungi are less recurrent than the other genera.
Received: 27 August 2021; Accepted: 11 February 2022
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Acknowledgements
Acknowledgments to the Foundation for Research Support in the State of Goiás - FAPEG, National Council for 
Scientific and Technological Development - CNPq, Evangelical Educational Association - AEE and Federal Insti-
tute of Goiás - IFGoiano. This research was funded in whole, or in part, by the Wellcome Trust [Grant number 
217968/Z/19/Z]. For the purpose of open access, the author has applied a CC BY public copyright license to any 
Author Accepted Manuscript version arising from this submission.
Author contributions
L.L.S.—Responsible for the execution of the projectR.N.A. advisor of master’s analysisM.J.B.—General coordi-
nator of the project, responsible for writingS.R.F.—Responsible for the soil laboratoryS.M.E.F.—responsible for 
spore analysisM.L.F., responsible for the field experimentX.E.G.—responsible for field guidanceS.J.M.—respon-
sible for colonization analysesD.eS.S.—Responsible for correcting the text.N.R.—Responsible for correcting the 
text.
Competing interests 
The authors declare no competing interests.
Additional information
Correspondence and requests for materials should be addressed to J.B.M.
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