Rhodosporidiobolus geoffroeae sp. nov., a basidiomycetous yeast isolated from the waste deposit of the attine ant Acromyrmex lundii.

A novel basidiomycetous yeast was isolated from the waste deposit of the attine ant Acromyrmex lundii (Hymenoptera: Formicidae). The field colony was located in Santurce town, Santa Fe province, Argentina. The description of the novel species was based on strain LLU043T. Analysis of the D1/D2 domains of the LSU rRNA gene sequences in GenBank demonstrated that strain LLU043T, belongs to the Rhodosporidiobolus clade and is closely related to Rhodosporidiobolus lusitaniae and Rhodosporidioboluscolostri with 97 % similarity to the two species. The novel species differs from R. lusitaniae and R. colostri in some physiological characteristics such as the lack of assimilation of cellobiose, salicin, succinate, citrate and ethylamine. The name Rhodosporidiobolus geoffroeae sp. nov. is proposed, with LLU043T (=CBS 12828T=CBMAI 1618T) as the type strain.

Wickerhamomyces spegazzinii sp. nov., an ascomycetous yeast isolated from the fungus garden of Acromyrmex lundii nest (Hymenoptera: Formicidae).

A novel ascomycetous yeast species in the genus Wickerhamomyces was isolated from the fungus garden of an attine ant nest, Acromyrmex lundii (Hymenoptera: Formicidae), from Santa Fe province, Argentina. Pairwise sequence alignment of D1/D2 sequences in the GenBank (http://www.ncbi.nlm.nih.gov) database revealed that the novel species is related most closely to Wickerhamomyces subpelliculosus, Wickerhamomyces linferdii, Wickerhamomyces anomalus, Wickerhamomyces siamensis and Wickerhamomycesciferrii with 96% similarity to the first four. The species name Wickerhamomyces spegazzinii sp. nov. is proposed to accommodate this novel strain, which differs from the above species in melibiose, 5-keto-D-gluconate, succinate, and DL-lactate assimilation among others. The type strain is JLU025T (=CBS 12756T=CBMAI 1619T).

Escovopsis trichodermoides sp. nov., isolated from a nest of the lower attine ant Mycocepurus goeldii.

Currently, five species are formally described in Escovopsis, a specialized mycoparasitic genus of fungus gardens of attine ants (Hymenoptera: Formicidae: tribe Attini). Four species were isolated from leaf-cutting ants in Brazil, including Escovopsis moelleri and Escovopsis microspora from nests of Acromyrmex subterraneus molestans, Escovopsis weberi from a nest of Atta sp. and Escovopsis lentecrescens from a nest of Acromyrmex subterraneus subterraneus. The fifth species, Escovopsis aspergilloides was isolated from a nest of the higher attine ant Trachymyrmex ruthae from Trinidad. Here, we describe a new species, Escovopsis trichodermoides isolated from a fungus garden of the lower attine ant Mycocepurus goeldii, which differs from the five other species by highly branched, trichoderma-like conidiophores lacking swollen vesicles, with reduced conidiogenous cells and distinctive conidia morphology. Phylogenetic analyses based on partial tef1 gene sequences support the distinctiveness of this species. A portion of the internal transcribed spacers of the nuclear rDNA was sequenced to serve as a DNA barcode. Future molecular and morphological studies in this group of fungi will certainly unravel the taxonomic diversity of Escovopsis associated with fungus-growing ants.

A Brazilian population of the asexual fungus-growing ant Mycocepurus smithii (Formicidae, Myrmicinae, Attini) cultivates fungal symbionts with gongylidia-like structures.

Attine ants cultivate fungi as their most important food source and in turn the fungus is nourished, protected against harmful microorganisms, and dispersed by the ants. This symbiosis evolved approximately 50-60 million years ago in the late Paleocene or early Eocene, and since its origin attine ants have acquired a variety of fungal mutualists in the Leucocoprineae and the distantly related Pterulaceae. The most specialized symbiotic interaction is referred to as "higher agriculture" and includes leafcutter ant agriculture in which the ants cultivate the single species Leucoagaricus gongylophorus. Higher agriculture fungal cultivars are characterized by specialized hyphal tip swellings, so-called gongylidia, which are considered a unique, derived morphological adaptation of higher attine fungi thought to be absent in lower attine fungi. Rare reports of gongylidia-like structures in fungus gardens of lower attines exist, but it was never tested whether these represent rare switches of lower attines to L. gonglyphorus cultivars or whether lower attine cultivars occasionally produce gongylidia. Here we describe the occurrence of gongylidia-like structures in fungus gardens of the asexual lower attine ant Mycocepurus smithii. To test whether M. smithii cultivates leafcutter ant fungi or whether lower attine cultivars produce gongylidia, we identified the M. smithii fungus utilizing molecular and morphological methods. Results shows that the gongylidia-like structures of M. smithii gardens are morphologically similar to gongylidia of higher attine fungus gardens and can only be distinguished by their slightly smaller size. A molecular phylogenetic analysis of the fungal ITS sequence indicates that the gongylidia-bearing M. smithii cultivar belongs to the so-called "Clade 1"of lower Attini cultivars. Given that M. smithii is capable of cultivating a morphologically and genetically diverse array of fungal symbionts, we discuss whether asexuality of the ant host maybe correlated with low partner fidelity and active symbiont choice between fungus and ant mutualists.

Association between Pseudonocardia symbionts and Atta leaf-cutting ants suggested by improved isolation methods.

Fungus-growing ants associate with multiple symbiotic microbes, including Actinobacteria for production of antibiotics. The best studied of these bacteria are within the genus Pseudonocardia, which in most fungus-growing ants are conspicuously visible on the external cuticle of workers. However, given that fungus-growing ants in the genus Atta do not carry visible Actinobacteria on their cuticle, it is unclear if this genus engages in the symbiosis with Pseudonocardia. Here we explore whether improving culturing techniques can allow for successful isolation of Pseudonocardia from Atta cephalotes leaf-cutting ants. We obtained Pseudonocardia from 9 of 11 isolation method/colony component combinations from all 5 colonies intensively sampled. The most efficient technique was bead-beating workers in phosphate buffer solution, then plating the suspension on carboxymethylcellulose medium. Placing these strains in a fungus-growing ant-associated Pseudonocardia phylogeny revealed that while some strains grouped with clades of Pseudonocardia associated with other genera of fungus-growing ants, a large portion of the isolates fell into two novel phylogenetic clades previously not identified from this ant-microbe symbiosis. Our findings suggest that Pseudonocardia may be associated with Atta fungus-growing ants, potentially internalized, and that localizing the symbiont and exploring its role is necessary to shed further light on the association.

Foraging of Psilocybe basidiocarps by the leaf-cutting ant Acromyrmex lobicornis in Santa Fé, Argentina.

BACKGROUND: It is generally accepted that material collected by leaf-cutting ants of the genus Acromyrmex consists solely of plant matter, which is used in the nest as substrate for a symbiotic fungus providing nutrition to the ants. There is only one previous report of any leaf-cutting ant foraging directly on fungal basidiocarps. FINDINGS: Basidiocarps of Psilocybe coprophila growing on cow dung were actively collected by workers of Acromyrmex lobicornis in Santa Fé province, Argentina. During this behaviour the ants displayed typical signals of recognition and continuously recruited other foragers to the task. Basidiocarps of different stages of maturity were being transported into the nest by particular groups of workers, while other workers collected plant material. CONCLUSIONS: The collection of mature basidiocarps with viable spores by leaf-cutting ants in nature adds substance to theories relating to the origin of fungiculture in these highly specialized social insects.

Association between Pseudonocardia symbionts and Atta leaf-cutting ants suggested by improved isolation methods

Fungus-growing ants associate with multiple symbiotic microbes, including Actinobacteria for production of antibiotics. The best studied of these bacteria are within the genus Pseudonocardia, which in most fungus-growing ants are conspicuously visible on the external cuticle of workers. However, given that fungus-growing ants in the genus Atta do not carry visible Actinobacteria on their cuticle, it is unclear if this genus engages in the symbiosis with Pseudonocardia. Here we explore whether improving culturing techniques can allow for successful isolation of Pseudonocardia from Atta cephalotes leaf-cutting ants. We obtained Pseudonocardia from 9 of 11 isolation method/colony component combinations from all 5 colonies intensively sampled. The most efficient technique was bead-beating workers in phosphate buffer solution, then plating the suspension on carboxymethylcellulose medium. Placing these strains in a fungus-growing ant-associated Pseudonocardia phylogeny revealed that while some strains grouped with clades of Pseudonocardia associated with other genera of fungus-growing ants, a large portion of the isolates fell into two novel phylogenetic clades previously not identified from this ant-microbe symbiosis. Our findings suggest that Pseudonocardia may be associated with Atta fungus-growing ants, potentially internalized, and that localizing the symbiont and exploring its role is necessary to shed further light on the association (AU)

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