Glomus halonatum Rose & Trappe (Glomeromycota) in South America: comments on the morphological characteristics of the species

Glomus halonatum Rose & Trappe (Glomeromycota) na América do Sul: comentários sobre a morfologia das espécies

ARTICLES

Glomus halonatum Rose & Trappe (Glomeromycota) in South America: comments on the morphological characteristics of the species

Glomus halonatum Rose & Trappe (Glomeromycota) na América do Sul: comentários sobre a morfologia das espécies

Bruno Tomio Goto; Cynthia Maria Carneiro Costa; Leonor Costa Maia¹

Departamento de Micologia, Universidade Federal de Pernambuco, 50670-420, Recife, PE, Brasil

RESUMO

Glomerosporos ornamentados de um fungo micorrízico arbuscular foram coletados em uma área de Floresta Atlântica em Goiana (Pernambuco, Brasil). A área tinha sido cultivada com Coffea canephora Pierre ex. Froehner [=Coffea robusta (L.) Linden)] e Bixa orellana L., abandonada, e hoje está coberta por espécies herbáceas invasoras. Após análise taxonômica usando critérios descritivos e terminológicos atuais, a espécie foi identificada como Glomus halonatum. São apresentados descrição, ilustrações e comentários sobre a espécie.

Palavras-chave:Glomeraceae, Glomus, diversidade, taxonomia

ABSTRACT

Ornamented glomerospores of an arbuscular mycorrhizal fungus were collected from an area of Atlantic Forest in Goiana (Pernambuco, Brazil). The area had been cultivated with Coffea canephora Pierre ex. Froehner [=Coffea robusta (L.) Linden)] and Bixa orellana L., then left fallow and today is covered by herbaceous raider species. After taxonomic analysis using current descriptive and terminological criteria, the species was identified as Glomus halonatum. A description, illustrations and comments about the species are presented.

Key words:Glomeraceae, Glomeromycota, Glomus, diversity, taxonomy

Introduction

Arbuscular mycorrhizal fungi are widely distributed in terrestrial ecosystems and can be found in both natural and agricultural areas. The documentation of new patterns of species distribution is necessary for the accurate estimation of the diversity and distribution of this important group of symbiotic fungi.

Some morphological characteristics used for the identification of AMF species, such as color and size of spores vary and can be affected by intrinsic or environmental factors (Bentivenga et al. 1997; Walker & Vestberg 1998). Thus, isolates of a single species collected from different localities can present variations in color and size, but, although serving as useful morphological support, these differences should not be used on their own to identify a species (Silva et al. 2005).

Descriptions of AMF species became more detailed when Walker (1983) proposed the use of terminology based on separation and types of spore walls. Today, with particular emphasis given to the ontogenesis of the spore wall layers most researchers have adopted this terminology (Morton 1995; Stürmer & Morton 1997, 1999; http://invam.caf.wvu.edu).

The purpose of this work was to record the occurrence of Glomus halonatum in Brazil, to describe, illustrate, and compare its description with those of similar species, providing additional information on the morphological aspects of this AMF. This species was earlier recorded in England and Mexico, where it was originally described (Rose & Trappe 1980) and, recently, in a list of AMF species found in the rhizosphere of Eucalyptus, in Bahia (Carrenho et al. 2008).

Material and methods

Soil samples were collected (0-20 cm deep) at random from an area of Atlantic Forest located at the Experimental Station of Itapirema (07º34'00"S; 35º00'00"W) belonging to IPA (Instituto Agronômico de Pernambuco), Goiana Municipality, Pernambuco state, Northeast Brazil). The area had been cultivated with Coffea canephora Pierre ex. Froehner [=Coffea robusta (L.) Linden], and Bixa orellana L., then left fallow, and today is covered by a variety of herbaceous raider species. The local soil has the following characteristics: 3 mg P dm^-3 of soil; pH in water 6.20; 0.09 (K), 0.00 (Al), 1.55 (Ca) and 0.60 (Mg) cmol_c dm^-3; 64% (coarse sand), 24% (fine sand) 2% (silt), and 10% (sandy loam).

Spores of AMF were extracted from soil by wet sieving and sucrose centrifugation (Gerdemann & Nicolson 1963; Jenkins 1964), separated and mounted with PVLG (alcohol polivinyl lactoglycerol) or PVLG + Melzer. Pot cultures were mounted using Panicum miliaceum L. and Sorghum bicolor L. as plant hosts, during two cycles of four months, though without success on spore multiplication. For the description of the species the terminology proposed by Morton was adopted (http://invam.caf.wvu.edu) and, for spores, that suggested by Goto & Maia (2006).

Results and discussion

Glomus halonatum Rose & Trappe Mycotaxon 10: 413, 1980.

Fig. 1, 2, 3 and 4.

Sporocarps not observed. Glomerospores usually globose (325-) 202.5×200 (-297.5) μm from yellow to orange brown (Fig. 1). Spore wall with three layers. The first evanescent, hyaline, mucilaginous, varying from 12.5–27.5 μm in thickness and in some cases presenting a tenuous Melzer's reaction (Fig. 2 and 3). This layer is normally smooth in young spores and granulose or with wide depressions at maturity. In senescent spores it can be absent or totally covered by soil debris. The second layer is laminated (17.5-25 µm), yellow brown to orange brown, with 1.0-2.4 µm tall spines on the external surface (Fig. 4). Occasionally this second layer can be divided, giving the impression that two laminated layers are present. The third layer is hyaline, with membranous appearance (< 2.0 µm) and apparently forms a septum in the insertion of the subtending hypha. One subtending hypha per spore, with a septum near the insertion, yellow to yellow brown wall (17.5-37.5 µm), formed by two layers: an external one continuous with the evanescent layer, hyaline to mucilaginous, and a laminated layer similar to that of the spore and ornamented. This ornamentation is observed for approximately 10 µm in the subtending hypha. No layer produces a distinct Melzer reaction, but an ill-defined reaction on the surface of the evanescent wall was observed. Germinal tube not observed.

Distribution: Mexico and England (Rose & Trappe 1980). In Brazil, spores were found in the rhizosphere of Eucalyptus in the state of Bahia (Carrenho et al. 2008) and isolated from soil at Goiana, Pernambuco state.

Examined Material: Pernambuco: Municipality of Goiana, G. halonatum Rose & Trappe. (X/2002), Costa, URM 45715; isolated from soil in area covered by herbaceous vegetation located at the Experimental Station of Itapirema GERMANY. Vogtsburg: Glomus spinuliferum Oehl & Sieverding, (VI/2002), Oehl, URM 45714 isotype, isolated from areas covered by semi-natural grassland in Vogelsang Pass, near Vogtsburg (Natural Reserve of Kaiserstuhl), Germany.

The morphological characteristics (color, size, and form and wall structure) of glomerospores of G. halonatum found in Pernambuco are similar to those described in the protologue (Rose & Trappe 1980), except for the absence of sporocarps and by the thickness of the first spore wall layer. In the examined material this layer was hyaline and evanescent and slightly thicker (12.5-27.5 µm) than that mentioned in the original description (8-20 µm).

Glomus halonatum can be mistaken for Glomus monosporum Gerdemann & Trappe due to the similarity of color and ornamentation of the glomerospores (Gerdemann & Trappe 1974), but the latter have only two wall layers, while G. halonatum has three spore wall layers.

Glomerospores of G. halonatum can also be mistaken for those of G. pansihalos Berch & Koske, G. spinosum Hu, G. verruculosum Blaszkowski and G. spinuliferum Sieverd. & Oehl. For example, Glomus pansihalos, G. spinosum and G. halonatum present the same number of spore wall layers and in all of them a hyaline, thick wall layer is present. However, in G. pansihalos the external layer presents radial striations in columns when in PVLG (Berch & Koske 1986), while in G. halonatum no striations are found. Moreover, G. halonatum presents homogenously distributed spine ornamentation, whereas G. pansihalos and G. spinosum present sparser ornamentation, similar to warts (Berch & Koske 1986; Hu 2002).

The glomerospores of G. halonatum have three wall layers whereas those of G. verruculosum have only two. Moreover, the latter have round tip ornamentation (warts) (Blaszkowski & Tadych 1997) whereas in G. halonatum the ornamentation has spiny tips.

Glomerospores of G. halonatum are similar to those of G. spinuliferum in color and spore wall structure. Both species have spores with three wall layers, one of which with spines immersed in the evanescent, hyaline and mucilaginous layer. However, G. halonatum has bigger spores than G. spinuliferum and sporocarps covered by a peridium of hypha loosely arranged (Rose & Trappe 1980), while in G. spinuliferum no sporocarps were observed at all (Oehl et al. 2003). Rose & Trappe (1980) mentioned one, two or three glomerospores loosely grouped in sporocarps of G. halonatum. The technique for extraction of spores from soil can disturb the sporocarps, contributing to the non-observation of such structures in the examined material. Up to now, spore cultures of G. halonatum and G. spinuliferum were not successful, so the descriptions are based on glomerospores isolated directly from the field (Rose & Trappe 1980; Oehl et al. 2003).

Glomus halonatum is also distinguished from G. spinuliferum by the thick evanescent layer that forms a halo similar to a ring in most spores. Oehl et al. (2003) stated that the external spore wall of G. spinuliferum (1-2 µm) can occasionally expand up to 50 µm in thickness in PVLG. This effect has also been described for Glomus pansihalos Berch & Koske, Glomus macrocarpum Tul. & Tul., Glomus coronatum Giovannetti and lately in Glomus caesaris Sieverding & Oehl (Berch & Koske 1986; Berch & Fortin 1983; Giovannetti et al. 1991; Oehl et al. 2002), as discussed by Morton (1986 a,b). It was not possible to observe the expansion of the external wall layer in glomerospores of G. spinuliferum probably due to the small number of spores present in the isotype examined. The absence of other distinctive characteristics shows the need for better examination of G. halonatum and G. spinuliferum isolates, to look for more differences between these species, considering that both are, in some aspects, morphologically similar.

In the same way, none of the species discussed in comparison to G. halonatum present this thick, evanescent wall layer, forming a distinct halo around the glomerospore. Thus, G. halonatum differs from all other Glomus species by presenting this distinctive morphological feature.

Acknowledgements

To Dr. Fritz Oehl, for providing slides of the isotype of G. spinuliferum, Dr. Reginaldo Silva, for helping with the photographs, and Dr. David Bousfield, for correcting the English text and making suggestions. The authors also acknowledge the following Institutions: Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Fundação de Apoio ao Desenvolvimento Científico e Tecnológico de Pernambuco (FACEPE) and Instituto Tecnológico de Pernambuco (ITEP) which provided, respectively, a research grant to L.C. Maia, a PhD scholarship to B.T. Goto and a post-doctoral scholarship to C.M.C. Costa.

Received: December 17, 2007

Accepted: May 11, 2009

• Bentivenga, S.P.; Bever, J.D. & Morton, J.B. 1997. Genetic variation of morphological characters within a single isolate of the endomycorrhizal fungus Glomus clarum (Glomaceae). American Journal of Botany 84: 1211-1216.
• Berch, S.M. & Fortin, J.A. 1983. Lectotypification of Glomus macrocarpum and proposal of new combinations: Glomus australe, Glomus versiforme, and Glomus tenebrosum (Endogonaceae). Canadian Journal of Botany 61: 2608-2617.
• Berch, S.M. & Koske, R.E. 1986. Glomus pansihalos, a new species in the Endogonaceae, Zygomycetes. Mycologia 78: 832-836.
• Blaszkowski, J. & Tadych, M. 1997. Glomus multiforum and G. verruculosum, two new species from Poland. Mycologia 89: 804 - 811.
• Carrenho, R.; Barbosa, F.F.; Araújo, C.V.M.; Alves, L.A. & Santos, O.M. 2008. Mycorrhizal Associations in Eucalyptus: Status and Needs. Tree and Forestry Science and Biotechnology 2: 57-67.
• Caproni, A.L.; Franco, A.A.; Berbara, R.L.L.; Gralha, J.R.D.O.; Ribeiro, E.M.S. & Sagin Junior, O.J. 2003. Capacidade infectiva de fungos micorrízicos arbusculares em áreas reflorestadas após mineração de bauxita no Pará. Pesquisa Agropecuária Brasileira 38: 937-945.
• Gerdemann, J.W. & Nicolson, T.H. 1963. Spores of mycorrhizal Endogone species extracted from soil by wet sieving and decanting. Transactions of the British Mycological Society 46: 235-244.
• Gerdemann, J.W. & Trappe, J.M. 1974. The Endogonaceae in the Pacific Northwest. Mycologia Memoir 5: 1-75.
• Giovanetti, M.; Avio, L. & Salutini, L. 1991. Morphological, cytological, and ontogenetic characteristics of a new species of vesicular-arbuscular mycorrhizal fungus. Canadian Journal of Botany 69: 161-167.
• Goto, B.T. & Maia, L.C. 2006. Glomerospores: a new denomination for the spores of Glomeromycota, a group molecularly distinct from the Zygomycota. Mycotaxon 96: 129-132.
• Hu, H.T. 2002. Glomus spinosum sp. nov. in the Glomaceae from Taiwan. Mycotaxon 83: 159-164.
• Jenkins, W.R. 1964. A rapid centrifugal-flotation technique for separating nematodes from soil. Plant Disease Reporter 48: 692.
• Moreira-Souza, M. Trufem, S.F.B. Gomes-da-Costa, S.M. & Cardoso, E.J.B.N. 2003. Arbuscular mycorrhizal fungi associated with Araucaria angustifolia (Bert.) O. Ktze. Mycorrhiza 13: 211-215.
• Morton, J.B. 1986a. Three new species of Acaulospora (Endogonaceae) from high aluminum, low pH soils in West Virginia. Mycologia 78: 641-648.
• Morton, J.B. 1986b. Effects of mountants and fixatives on wall structure and Melzer's reaction in spores of two Acaulospora species (Endogonaceae). Mycologia 78: 787-794.
• Morton, J.B. 1995. Taxonomic and phylogenetic divergence among five Scutellospora species (Glomales, Zygomycetes) based on comparative developmental sequence. Mycologia 87: 127-137.
• Oehl, F.; Wiemken, A. & Sieverding, E. 2002. Glomus caesaris, a new arbuscular mycorrhizal fungus from the Kaiserstuhl in Germany. Mycotaxon 84: 379-385.
• Oehl, F.; Wiemken, A. & Sieverding, E. 2003. Glomus spinuliferum, a new ornamented species in the Glomales. Mycotaxon 86: 157-162.
• Rose, S.L. & Trappe, J.M. 1980. Three new endomycorrhizal Glomus spp. associated with actinorrhizal shrubs. Mycotaxon 10: 413-420.
• Silva, G.A.; Maia, L.C. & Stürmer, S.L. 2005. A dichotomous key to Scutellospora species (Gigasporaceae, Glomeromycota) using morphological characters. Mycotaxon 94: 293-301.
• Stürmer, S.L. & Morton, J.B. 1997. Developmental patterns defining morphological characters in spores of four species in Glomus Mycologia 89: 72-81.
• Stürmer, S.L. & Morton, J.B. 1999. Taxonomic reinterpretation of morphological characters in Acaulosporaceae based on developmental patterns. Mycologia 91: 849-857.
• Walker, C. 1983. Taxonomics concepts in the Endogonaceae: Spore wall characteristics in species descriptions. Mycotaxon 18: 443-455.
• Walker, C. & Vestberg, M. 1998. Synonymy amongst the arbuscular mycorrhizal fungi: Glomus claroideum, G. maculosum, G. multisubstensum and G. fistulosum Annals of Botany 82: 601-624.

Publication Dates

• Publication in this collection
  07 Apr 2010
• Date of issue
  Dec 2009

History

• Received
  17 Dec 2007
• Accepted
  11 May 2009