IJAAR 4 (2016) 6-9 ISSN 2053-1265

Acaulospora excavata () in agricultural soils of Chiapas, México

Pérez -Luna Y.1*, Alvarez-Solís D.2, Hernández Cuevas L.3 and Sánchez-Roque Y.1

1Universidad Politécnica de Chiapas, Eduardo J. Selvas s. n. Col. Magisterial. C.P. 29080, Tuxtla Gutiérrez, Chiapas, México. 2El Colegio de la Frontera Sur (CONACYT), Carretera Panamericana y Periférico Sur s/n. C. P. 29290, San Cristóbal de Las Casas, Chiapas, México. 3Universidad Autónoma de Tlaxcala. km 10 Carretera Texmelucan-Ixtacuixtla, Tlaxcala 90122, Tlaxcala, México.

Article History ABSTRACT Received 02 February, 2016 In the State of Chiapas, Mexico, one of the most biodiverse country, there have Received in revised form 18 been few studies to determine the richness of arbuscular mycorrhizal fungi and February, 2016 Accepted 22 February, 2016 to date there have been only 24 . The aim of this study was to record the presence of excavate (a species recorded for few locations in Key words: Mexico) in agricultural plots of corn, adjacent to Mopri Forest Reserve in Ivory Hongos Coast. This finding was the result of a study conducted in diverse fields of Ejido micorrizógenos La Bella Illusion, where more than 22 species of Glomeromycota, belonging to arbusculares (HMA), the genera Acaulospora, Entrophospora, Gigaspora, Glomus and Archaeospora Espore, were identified. Inoculum.

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INTRODUCTION

Arbuscular mycorrhizal fungi (AMF) are soil micro- most biologically rich region (González-Espinosa et al., organisms that form symbiotic associations with plant 2005). Studies focused on AMF, evaluated the effect of roots, increasing the amount of nutrients absorbed by the these fungal symbiotic associations with crops of plant, especially phosphorus (P), copper (Cu) and zinc economic importance to the State (Adriano et al., 2011; (Zn), they are considered as biological resource because Pérez-Luna et al., 2012; Díaz-Hernández et al., 2013). In they generate environmental benefits by improving the the last 20 years, approximately 200 species of AMF physicochemical and biological soil conditions have been classified into four orders: Glomerales, (Karagiannidis et al., 2011). Due to the fact that the Diversisporale, Paraglomerales and Archaeosporales benefits generated by these fungi are increasingly (Schüßler and Walker, 2010). However the diversity of important, it is therefore necessary to identify them in these microorganisms could be higher than those already different soils, creating the opportunity to isolate and recognized (Öpik et al., 2008). In natural ecosystems, it identify different AMF (Zenget al., (2013). has been found that the diversity of AMF is very high The taxonomic diversity of AMF in Chiapas, México, (Schüβlera et al., 2001; Öpik et al., 2006; Guadarrama, has been little studied, even though this is one of the 2008). But diversity of AMF in farming systems decreases due to agricultural practices such as tillage which destroys the hyphal network in the soil (Evans and Miller, 1990; Jansa et al., 2002; Schalamuk et al., 2006; *Corresponding author. E-mail: [email protected]. Tel: Lovera and Cuenca, 2007; Schalamuk and Cabello, +52 (961) 6120484. Fax: +52 (961) 6120499. 2009). The diversity of AMF in México is recorded in

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Varela and Trejo (2001) who reported the presence of and isolated according to Gerdemann and Nicholson and Funneliformi sconstrictus (1963). Later the spores were mounted in lactoglycerol (Glomus constrictum), in sugarcane soils and coconut, polyvinyl (PVLG) with or without Melzer’s reagent for respectively in Chiapas. However, for this entity, in recent identification under microscope (Méndez-Cortés et al., years 44 species of AMF from areas of corn fields, pine 2011; Schenck and Pé rez, 1990). forests were recorded and mountain cloud forest (Violi et The spore size and wall thickness and layers were al., 2008), and only 23 species of corn field (Pérez-Luna measured using a calibrated micrometer. et al., 2012); demonstrating that the diversity of AMF in Taxonomic identification was performed by comparing Chiapas can be very high and that it is little explored the morphological characteristics observed in the present considering the great diversity of natural and agro- study with those described for Acaulospora spp. that ecosystems (González-Espinosa et al., 2005). produce spores with ornamented walls (Taylor et al., , the species reported in this 2014). Permanent preparations spores which is located in research, was described for the first time in soil of Agro-industrial Engineering Laboratory of the Universidad Terminalia superb and of the Mopri Politécnica de Chiapas, Tuxtla Gutierrez, Chiapas, Forest Reserve, Côte d'Ivoire, West Africa (Ingleby et al., México. 1994) and it has only been reported from Mexican grasslands, tropical forest and agroforestry systems in the region of the Tuxtlas (Varela et al., 2008) and coffee RESULTS AND DISCUSSION growing areas and mountain cloud forest in central Veracruz (Heredia and Arias, 2008; Arias et al., 2011), of A. excavata is one of 23 AMF species present in the avocado orchards in Michoacan (Bárcenas-Ortega et al., rhizosphere of corn in two places where the conditions of 2011) and now areas of cornfield in the State of Chiapas. agricultural management and vegetation are very The aim of this study was to show the presence of A. different, one of them the corn is grown intercropping excavata in soils cultivated with corn in Chiapas. Nescafé bean (Mucuna deeringiana Merr.), while in the other there is no cover, in both plots it was driven rotation corn common bean (Phaseolus spp.).The Nescafé bean MATERIALS AND METHODS is planted between February and March, when it reaches its vegetative development is broken and is left as ground The experiment was established in the Ejido La Bella cover where corn is planted. The soils are clay texture, Ilusión, located in the City of Tenejapa, Chiapas, México, pH 6.3-6.5, organic matter content of 6.0-6.6% total at an altitude of 400 meters above sea level. The nitrogen concentration of 0.30-0.33%, phosphorous total geographic location is latitude 16° 09’ 45” North and concentration of 5.3-5-6 mg kg-1. Other AMF species longitude 91° 13’ 10”, corresponding to tropical regions, found in this study belong to the Glomus and with more than 2500 mm annual rainfall (Figure 1A), with Acaulospora such as Glomus claroideum, Glomus the objective of evaluating the AMF diversity associated geosporum, Glomus intrarradices, Glomus aff. with the cultivation of corn in organic farming, 28 samples lamellosum, Glomus luteum, Glomus sinuosum, Glomus were collected, each made up of 15 sub-samples of corn verruculosum, , Acaulospora spinosa, rhizosphere soil at a distance of 5 cm from corn plants A. scrobiculata, as well as Entrophospora infrequens, and 15 cm deep; physical and chemical attributes of the Gigaspora gigantea and Intraspora schenkii, to name but soil were determined, such as pH (relation 1:2 with H2O), a few. organic matter (method of Walkley and Black), Total The spores of Acaulospora found in the soils of the nitrogen (semi-micro Kjeldahl method) and total explored fields, are ornamented with large and prominent phosphorus (Olsen method). depression. The spores isolated during this study presented morphological characteristics similar to those described by Ingleby et al. (1994), who described the Characterization of the rhizosphere soil species as A. excavata. The spores are present individually in the soil, Edocher to The hongos micorrizógenos arbusculares (HMA) was orange, and 150-170 μm in diameter. Some spores obtained from rhizosphere soil, defined as that part of the possessed the remains of sporiferous saccule adhered to soil that is influenced by the roots, where the root them, in this case the saccule was not observed; which exudates affect processes soil and microorganisms found may be due to mature spores collapses. Spores of A. in it, of the Ejido La Bella Ilusión; this is located in the excavata possess a distinctly ornamented outer wall municipality of Tenejapa, Chiapas, México. surface of densely crowded, flat-bottomed pits; deep The characterization of the rhizosphere soil of the 420 depressions by way of wells, of 5 μm deep, while that in sub-samples allows obtaining the number of spores of the surface of the spore are observed plates together. AMF g-1 rhizosphere soil. The spores were extracted In A. excavata are observed three walls, a laminated

Pérez-Luna et al. 8

A

B C

Figure 1. A. excavata spores isolated from the rhizosphere of corn. A) Plots of Ejido “La Bella Illusion”, Chiapas, México; B) Reaction with Melzer in the last stratum (indicated by an arrow); C) Detail of ornamentation in the laminated layer (Melzer reagent assembly).

brown-pale wall, which does not react with the reactive (Varela et al., 2008), such as coffee plantations of the Melzer in which circular holes are observed with 5 μm Tuxtlas, Veracruz, with different plant structure and deep, surrounded by an angled edge; followed by a intensity of handling (Arias et al., 2011). Recently, Pereira single-walled, hyaline, membranous, and finally an et al. (2015) reported their occurrence in the semi-humid amorphous wall which reacts with Melzer and changes its Atlantic rainforest and semi-arid Caatinga biomes of color from hyaline to deep purple (Figure1B and C). North-eastern Brazil.

Conclusion REFERENCES

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