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Article

Generation of Nutrients and Detoxification: Possible Roles of Yeasts in Leaf-Cutting Ant Nests

by
Thais D. Mendes
1,
André Rodrigues
2,
Ifeloju Dayo-Owoyemi
2,3,
Fernando A. L. Marson
3 and
Fernando C. Pagnocca
2,3,*
1
EMBRAPA—Agroenergy/Parque Estação Biológica, Brasília, DF 70770-901, Brazil
2
Department of Biochemistry and Microbiology, UNESP—São Paulo State University, Rio Claro, SP 13506-900, Brazil
3
Centre for the Study of Social Insects, UNESP—São Paulo State University, Rio Claro, SP 13506-900, Brazil
*
Author to whom correspondence should be addressed.
Insects 2012, 3(1), 228-245; https://doi.org/10.3390/insects3010228
Submission received: 14 January 2012 / Revised: 12 February 2012 / Accepted: 13 February 2012 / Published: 17 February 2012
(This article belongs to the Special Issue Symbiosis: A Source of Evolutionary Innovation in Insects)
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Abstract

:
The possible roles played by yeasts in attine ant nests are mostly unknown. Here we present our investigations on the plant polysaccharide degradation profile of 82 yeasts isolated from fungus gardens of Atta and Acromyrmex species to demonstrate that yeasts found in ant nests may play the role of making nutrients readily available throughout the garden and detoxification of compounds that may be deleterious to the ants and their fungal cultivar. Among the yeasts screened, 65% exhibited cellulolytic enzymes, 44% exhibited pectinolytic activity while 27% and 17% possess enzyme systems for the degradation of protease and amylase, respectively. Galacturonic acid, which had been reported in previous work to be poorly assimilated by the ant fungus and also to have a negative effect on ants’ survival, was assimilated by 64% and 79% of yeasts isolated from nests of A. texana and Acromyrmex respectively. Our results suggest that yeasts found in ant nests may participate in generation of nutrients and removal of potentially toxic compounds, thereby contributing to the stability of the complex microbiota found in the leaf-cutting ant nests.

1. Introduction

Ants in the tribe Attini maintain a mutualistic association with basidiomycetous fungi cultivated for food [1]. The phylogenetically derived genera in this tribe, Atta and Acromyrmex, known as the leaf-cutting ants, cut and collect fresh leaves and flower parts as substrate for the cultivation of mutualistic fungi [1,2]. This fungus is responsible for the production of extracellular enzymes that help to breakdown the plant substrate found in the fungus gardens [3,4,5,6]. In this process, simple sugars and other nutrients are generated and accumulated in the fungus gardens and it is thought these may in part support development of workers [7].
Because ants provide an optimum environment for fungal growth, additional alien microbes may also find the fungus garden a suitable substrate for development. Thus, it is fundamental for the ants to protect the mutualistic fungus from potential harmful microorganisms. For this reason ants employ several mechanisms such as: (i) massive inoculation of the cultivar onto fresh collected plant substrate [8]; (ii) weeding and grooming behaviors to remove harmful fungi [9]; (iii) secretion of antimicrobial compounds by workers [10,11,12,13,14]; (iv) the use of antibiotic-producing actinobacteria [15,16,17,18,19].
Despite the hygienic strategies adopted by ants to keep their gardens free of alien microbes, members of the fungal genus Escovopsis are frequently found in attine gardens. Escovopsis is considered a specialized parasite that attacks the cultivar hyphae [20] and long-term infections occasionally drive the colony to death [2]. Furthermore, colonies infected with Escovopsis show lesser garden biomass and lower number of workers and brood than uninfected colonies, thus indicating the negative effects of this parasite in the symbiosis [2]. In addition to Escovopsis, several other microorganisms are found in this symbiosis [21,22,23,24,25,26,27,28,29,30,31,32,33,34]. It is likely that many microbes found in the fungus gardens may not play significant roles, but some may provide unknown but important functions.
Several authors have reported the occurrence of yeasts in attine gardens. Craven et al. [22] studied the fungus garden of Atta cephalotes and Acromyrmex octospinosus and reported the first occurrence of yeasts in attine ant nests. Angelis et al. [23] isolated yeasts in gardens of Atta sexdens and Atta laevigata. Carreiro et al. [24] isolated the yeast genera Candida, Cryptococcus, Rhodotorula and Trichosporon from laboratory nests of Atta sexdens rubropilosa. Black yeasts were also isolated from different parts of the ant’s integument [25,26,27]. Rodrigues et al. [28] isolated yeasts from fungus garden of A. texana and observed inhibition of the growth of Escovopsis and alien filamentous fungi, suggesting that they may also be involved in defending the fungus garden. In addition, three new yeast species namely Sympodiomyces attinorum [32], Cryptococcus haglerorum [33] and Trichosporon chiarellii [34] were described from this environment.
Preliminary studies suggest that bacteria may act as co-participants with the mutualistic fungus, in the degradation of the plant substrate. Ribeiro [29] and Bacci et al. [30] isolated bacteria from fungus garden of Atta sexdens and found strains that exhibited cellulases, pectinases, amylases and proteases. Recently, Suen et al. [35] revealed that the fungus garden contain a diverse community of bacteria with high lignocellulose-degrading capacity. Pinto-Tomás et al. [36] reported that bacteria in the fungus garden may fix nitrogen and it seems to be an important contribution for the ants. The genus Klebsiella was found as one of the most effective in this task.
To investigate the potential of yeasts as co-participants in the degradation of the plant material, we carried out a preliminary screening of cellulase, pectinase, amylase, xylanase and protease enzymes exhibited by yeasts associated with fungus gardens of leaf-cutting ants. By assessing the enzymatic profile of garden yeasts, we can begin to determine the putative contribution of these microorganisms in the degradation of the main plant polysaccharides, protein material and in the assimilation of its hydrolysis products. We show that the yeast community found in fungus gardens may play more important roles than previously thought.

2. Experimental Section

2.1. Yeast Isolates and Identification

We screened a total of 82 yeasts recovered from eight leaf-cutting ant species. Forty-four yeast strains were isolated and identified by Rodrigues et al. [28] from the fungus garden of Atta texana in Texas, USA. In addition, 38 isolates were recovered from 18 nests of several Acromyrmex species namely Acromyrmex ambiguus (n = 3 nests), Ac. coronatus (2), Ac. disciger (1), Ac. heyeri (3), Ac. laticeps (4), Ac. lundi (3), Ac. subterraneus (1)and Acromyrmex sp. (1) collected in south Brazil (Table S1). Samples from fungus gardens of Acromyrmex were processed as indicated in Rodrigues et al. [28] but using YMA medium without rose bengal as growth restrictor.
Yeasts isolated from Acromyrmex were first grouped based on phenotypic characteristics. Then, genomic DNA was extracted according to Almeida [37] and subjected to microsatellite-primed PCR (MSP-PCR) with primer (GTG)5, following the method of Sampaio et al. [38]. The D1/D2 domains of 26S rDNA of representatives strains (Table S2) were amplified using primers NL1 and NL4 [39]. Reactions were composed of 8.3 μL of Milli-Q water, 2.0 μL of each primer (10 µM), 2.5 μL of 10× buffer, 1.0 μL of MgCl2 (50 mM), 4.0 μL of dNTPs (1.25 mM each), 0.2 μL of Taq polymerase (5 U/μL) and 5.0 μL of diluted DNA templates (1:750). The reaction conditions were: 96 °C for 3 min, followed by 35 cycles at 96 °C for 30 s, 61 °C for 45 s and final extension at 72 °C for 1 min. Amplicons were purified with the Illustra GFX PCR DNA and Gel Band Purification Kit (GE Healthcare). Forward and reverse sequences were generated using the same amplification primers and using the BigDye Terminator kit on ABI 377 and ABI 3100 (Life Technologies).
Sequences were assembled in contigs and manually edited using BioEdit v.7.0.5.3 [40]. Contigs were queried at the NCBI-GenBank database (National Center for Biotechnology Information) using BLASTn algorithm [41]. Sequences derived from yeast isolated from Acromyrmex were deposited at GenBank (JQ317161-JQ317168). Sequence accessions for yeasts isolated from A. texana are provided in [28].

2.2. Assaying Yeasts for Hydrolytic Enzymes

Screenings for cellulase, pectinase, amylase, xylanase and protease enzymes were carried out qualitatively on different media supplemented with a specific polymer as described below. All plates were incubated at 25 °C for seven days for the detection of amylase, protease, and pectinase and 7–14 days for cellulase and xylanase. The formation of hydrolysis halos around the colonies indicated the production of enzymes and consequent degradation of polymers (see Figure S1 for representative examples of halo formation).
Starch hydrolysis was evaluated using the basal medium I described by Looder [42] supplemented with 20 g·L−1of soluble starch (Mallinckrodt). After incubation, plates were stained with lugol. For the assessment of CMCellulase, pectinase and xylanase a basal medium composed of 6.7 g·L−1 of YNB (Yeast Nitrogen Base, Difco) and 18 g·L−1 of agar supplemented with 5 g·L−1 of carboxymethylcellulose, 10 g·L−1 of polygalacturonic acid or 10 g·L−1 of xylan was used, respectively. The plates were stained with Congo red for CMCellulase [43], ruthenium red for pectinase [44] and lugol for xylanase [45]. Protein hydrolysis was assessed in solid medium containing 100 g·L−1 of Skim Milk medium (Difco). All polymers were supplied by Sigma Chemical Company.

2.3. Assimilation of the Hydrolysis Products

In addition to the hydrolytic enzymes, we evaluated the ability of yeasts to assimilate the simple sugars and products resultant of plant polysaccharides hydrolysis: maltose, cellobiose, galacturonic acid and xylose (Sigma Chemical Company). The determination was carried out on solid media containing 6.7 g·L−1 of YNB and 18 g·L−1 of agar supplemented with 5 g·L−1 of each carbon source. The growth of colonies on the simple sugars was compared with negative (medium without any carbon source) and positive controls (medium supplemented with glucose).

3. Results and Discussion

Several studies reported the occurrence of yeasts in the fungus gardens of both lower and higher attine ants [22,23,24,25,26,27,28,32,33,34,46]. Yeasts may enter the fungus gardens by the foraging activity of workers; specifically for leaf-cutting ants, yeasts may derive from the fresh plant material collected by workers as the phylloplane is considered a rich source of yeast species [47].
Degradation profile and assimilation of hydrolysis products by 82 yeasts isolated from Atta texana and Acromyrmex are shown in Table 1 and Table 2, respectively. Specifically for yeasts isolated from fungus gardens of A. texana, 77% showed at least one of the studied enzymes. Among the positive strains, six exhibited only one enzyme whereas the three Cryptococcus laurentii isolates exhibited four enzymes (Table 1 and Table S2 for results of specific isolates). Cryptococcus flavus and Pseudozyma sp. were the only isolates positive for all enzymes evaluated (Table 1 and Table S2 for results of specific isolates). Xylanase was exhibited by 59% of the isolates, amylase, CMCellulase, pectinase and protease activities were observed in 25%, 43%, 20% and 32% of the isolates, respectively (Figure 1). All the tested yeast isolates assimilated maltose, cellobiose and xylose and 63% assimilated galacturonic acid (Table 1).
Except for xylanolytic activity, which was not evaluated for yeasts isolated from Acromyrmex, all isolates exhibited at least one of the evaluated enzymes (Table 2). Amylase, CMCellulase, pectinase and protease activities were observed in 8%, 89%, 71% and 16% of the isolates, respectively (Figure 1). All yeasts isolated from Acromyrmex assimilated xylose and approximately 61%, 61% and 79% assimilated maltose, cellobiose and galacturonic acid, respectively (Table 2).
Our results indicated that the yeast species composition differed between gardens of A. texana and the various Acromyrmex (Table 1 and Table 2). Cryptococcus was the only genus shared by both ant genera. This yeast genus is widely distributed in the environment and has been shown to be prevalent among the yeast communities found on the phylloplane [47,48,49], but is also commonly found in soils from grasslands, pastures, and tundra [49]. Because of its widespread occurrence, Cryptococcus is expected to be in contact with workers and be transported to nests through the plant material collected by workers [24]. Previous studies on attine ants also recorded this genus as member of the microbiota of attine ants [24,25,33,34,46].
Table
Table 1. Enzymatic activity and assimilation profile of yeasts and yeast-like fungi isolated from fungus gardens of Atta texana.
Table 1. Enzymatic activity and assimilation profile of yeasts and yeast-like fungi isolated from fungus gardens of Atta texana.
Yeast Species 3N 4Hydrolytic Enzymes 1 Assimilation 2
AMCELPXYLPRT MCAX
Aureobasidium pullulans3-3 5-32 3323
Bullera sinensis111-1- 1111
Bulleromyces albus1-1-1- 1111
Candida membranifaciens3----- 33-3
Candida melibiosica1----- 11-1
Cryptococcus laurentii33212- 3333
Cryptococcus flavus333132 3333
Cryptococcus magnus3-3-22 33-3
Cryptococcus luteolus1----1 1111
Cryptococcus podzolicus11--1- 1111
Cryptococcus flavescens1---1- 1111
Cryptococcus cf. cellulolyticus2-2-2- 2222
Cryptococcus sp. (ATT178)1---1- 1111
Cryptococcus sp. 1 (ATT079)1-1-1- 11-1
Cryptococcus sp. 2 (ATT080)1----- 11-1
Cryptococcus sp. 3 (ATT123)1----- 1111
Cryptococcus sp. 4 (ATT176)11-11- 1111
Farysizyma sp. 61---11 1111
Hannaella kunmingensis 61----- 1111
Kodamaea ohmeri 1----- 11-1
Pseudozyma sp.111111 1111
Rhodotorula nothofagi11-11- 11-1
Rhodotorula mucilaginosa1--1-- 1111
Rhodotorula lactosa1----- 1111
Rhodotorula sp. HB 12111----- 1111
Sporidiobolus ruineniae2--2-- 22-2
Sporisorium penniseti (yeast-like)2--112 2222
Sympodiomycopsis paphiopedili2-1-21 22-2
unidentified yeast-like fungus2-1-12 2222
Total44111992614 44442844
1 AM: Amylase; CEL: CMCellulase; P: pectinase; XYL: xylanase; PRT: protease; -: negative results; 2 M: maltose; C: cellobiose; A: galacturonic acid; X: xylose; -: negative results; 3 GenBank accession were provided in Rodrigues et al. [28]; 4 N: Total number of yeast isolates in each species; 5 Figures indicate the number of positive isolates for a particular test; 6 These strains were previously identified in Rodrigues et al. [28] as Rhodotorula cf. taiwaniana and Cryptococcus cf. luteolus and now are confirmed to belong to Farysizyma sp. and H. kunmingensis, respectively.
Table
Table 2. Enzymatic activity and assimilation profile of yeasts isolated from fungus gardens of Acromyrmex.
Table 2. Enzymatic activity and assimilation profile of yeasts isolated from fungus gardens of Acromyrmex.
Yeast SpeciesN 4Closest Relative 1 Hydrolytic Enzymes 2 Assimilation 3
%Accession AMCELPPRT MCAX
Cryptococcus laurentii1100JQ317168 ---1 5 1111
Galactomyces candidus15100JQ317161 -15122 001415
Meyerozyma caribbica1100JQ317162 -1-- 1111
Meyerozyma guilliermondii6100JQ317164 -661 6636
Trichosporon chiarellii6100JQ317165 -341 6666
Trichosporon montevideense6100JQ317167 1621 6636
Trichosporon multisporum3100JQ317166 233- 3323
Total38 334276 23233038
1 According to BLASTn (NCBI-GenBank) results; %: Percent identity to sequences deposited at GenBank; 2 AM: Amylase; CEL: CMCellulase; P: pectinase; XYL: xylanase; PRT: protease; -: negative results; 3 M: maltose; C: cellobiose; A: galacturonic acid; X: xylose; -: negative results; 4 N: Total number of yeast isolates in each species; 5 Figures indicate the number of positive isolates for a particular test.
Insects 03 00228 g001 1024
Figure 1. Proportion of hydrolytic yeast isolates recovered from fungus gardens of the leaf-cutting ant genera Acromyrmex (n = 38 yeast isolates) and Atta (n = 44 isolates). For details check Table 1 and Table 2. Xylanase activity was not determinate for yeasts recovered from Acromyrmex.
Figure 1. Proportion of hydrolytic yeast isolates recovered from fungus gardens of the leaf-cutting ant genera Acromyrmex (n = 38 yeast isolates) and Atta (n = 44 isolates). For details check Table 1 and Table 2. Xylanase activity was not determinate for yeasts recovered from Acromyrmex.
Insects 03 00228 g001
Interestingly, Trichosporon chiarellii [34] was found in three nests, one of Ac. heyeri and two of Ac. lundi located at different sites (Table S1). This species was described from fungus gardens of Myrmicocrypta camargoi, a recently described lower attine [50]. So far, there are no reports of the isolation of T. chiarellii from sources other than attine gardens.
Although differences between the yeast communities from the target ant genera were evident, the degradation profile of plant polysaccharides by yeasts was similar between the two communities (Figure 1). In nature, yeasts are associated with specific habitats [51]; one might expect that their presence and abundance in gardens of attine ants may not be coincidental. The acidic pH and nutrient availability [52] may allow some yeast species to be selected and multiply in attine gardens. Thus, species found in such environment rich in readily available nutrients, may not only use the nutrients for their development, but also to exhibit enzymes that aid in the hydrolysis of the plant tissues, resulting in increased nutrient availability for the microbiota and facilitating substrate colonization by the mutualistic fungus.
Pectinolytic enzymes seem to play a key role in the fungus gardens because the mutualistic fungus of leaf-cutting ants exhibit such enzymes in large quantities [3,4]. The mutualistic fungus of Atta sexdens rubropilosa exhibits pectinase 7 times higher than amylase and more than 24 times higher than xylanase and 38 times higher than CMCellulase [3]. Pectinolytic enzymes are important because they promote the degradation of the pectic substances present in the middle lamella of plant cells, facilitating the colonization of the plant substrate and access to other cell wall components [53]. This seems to explain the large amounts of pectinolytic enzymes exhibited by the mutualistic fungus.
The proportion of yeasts that degrade pectin was 20% and 71% of isolates recovered from A. texana and Acromyrmex, respectively (Table 1 and Table 2). This would result in a further increase in pectin breakdown in the garden. However, despite the large amount of pectinolytic enzymes exhibited by the fungus cultivar, it does not present good growth in the presence of galacturonic acid, the most important hydrolysis product of pectin [3]. In addition, Silva et al. [7] experimentally demonstrated that galacturonic acid does not contribute to the nutrition of ants, but rather decrease the survival of Atta sexdens workers. Thus, it is expected that galacturonic acid would accumulate in the fungus gardens. In contrast to what was observed for the mutualistic fungus and workers, galacturonic acid was assimilated by 64% (Table 1) and 79% (Table 2) of the yeasts isolated from nests of A. texana and Acromyrmex, respectively. In addition, Carreiro [54] working with fungus gardens and refuse dumps of laboratory nests of A. sexdens rubropilosa recovered a very different yeast community profile when compared to our findings. However, considering these distinct communities, 22% of the isolates (n = 93 yeasts) found in Carreiro’s study also assimilated galacturonic acid. Thus, the results of two independent studies suggest that yeasts may perform a detoxification (filtering) process in attine gardens by consuming galacturonic acid. It was also observed that the assimilation of galacturonic acid was not restricted to a specific group of yeast; instead, this ability was distributed among the majority of the taxa (Table 1, Table 2 and Table S2). Thus, the utilization of galacturonic acid may be an advantage for the survival of yeasts recently introduced into the gardens and at same time it would be a remarkable contribution to the maintenance of the nest homeostasis.
An additional possible contribution of yeasts is the degradation of cellulose. This polysaccharide is the most abundant component of plant cell wall [55]. It was formerly assumed that cellulose was the main source of carbohydrates for the ant cultivar [56]. This assumption was reinforced by Bacci et al. [57], who showed that the mutualistic fungus exhibits cellulolytic enzymes. However, further studies demonstrated that the ability of the mutualistic fungus to degrade cellulose is limited when compared with other polysaccharidases as pectinases, amylases and xylanases [3,4]. In addition, Abril and Bucher [58] suggested that the mutualistic fungus does not exhibit cellulase, showing that the cellulolytic activity by the cultivar does not seem to be relevant. Recently, Suen and colleagues [35] determined the amount of cellulose present in the fungus gardens. The authors discovered low amounts of cellulose in mature garden parts when compared to younger portions where newly plant material is added by the ants, suggesting that cellulose degradation does occur in the fungus gardens. Using cultivation-independent techniques, the authors also showed that the fungus garden harbors a community of cellulose-degrading bacteria [35].
Our data shows that yeasts can also contribute to the degradation of cellulose present in the fungus gardens. The CMCellulose was degraded by 43% and 89% of the yeasts isolated from A. texana and Acromyrmex, respectively (Table 1 and Table 2). These data indicate that yeasts associated with fungus gardens may contribute, along with cellulose-degrading bacteria [35], to the production of easily assimilated sugars, especially glucose, for other members of the symbiosis, and also facilitate the colonization of plant tissues by the mutualistic fungus. On the other hand, cellobiose, the disaccharide resulting from the hydrolysis of cellulose, was assimilated by all yeast isolates from Atta texana (Table 1) and by 61% of yeasts from Acromyrmex (Table 2),revealing that this sugar may support the growth of yeasts. Also, there is evidence that Atta sexdens workers cannot utilize this sugar for survival [7] and the mutualistic fungus only grows at an intermediate rate on this carbon source [3]. Thus, cellobiose resulting from cellulose breakdown would be available for yeasts, thereby providing for their survival without competing with the other members of the symbiosis as also observed in respect to galacturonic acid.
The yeast participation in the generation of assimilable compounds from starch seems to be as important as that observed for the other plant polysaccharides. We found that 25% of the yeasts isolated from gardens of A. texana exhibited amylase, while only 8% of yeasts isolated from Acromyrmex exhibited this enzyme. The hydrolysis products of starch are mainly glucose and maltose. Glucose is the carbon source that best supports the development of the mutualistic fungus [3] and the survival of workers of Atta sexdens [7]. According to Silva et al. [59] glucose resultant from the degradation of starch by the ant cultivar is present in the fungus garden. Although the proportion of yeast capable of starch degradation is not high, they may also contribute to the hydrolysis of starch in gardens, generating easily assimilable sugars for their growth and the other members of the symbiosis. On the other hand, except for Galactomyces candidus (differentiated from G. geotrichum by growth at 35 °C according to [60]) all yeast species assimilated maltose (Table 1 and Table 2). These data indicate that the degradation of starch is essential for the maintenance of workers [7,59] and is also important for the yeasts present in nests.
Few studies have explored the use of nitrogen sources by the ants and their associated microbes in comparison to carbon sources. Abril and Bucher [61] reported that the ant cultivar grows on inorganic sources of nitrogen, but does not grow in media containing protein (peptone) as a nitrogen source. In addition, Silva et al. [7] reported that workers of Atta sexdens are not able to use peptone and suggest that ants are not able to feed on plant proteins. Recently, Pinto-Tomás et al. [36] observed nitrogen enrichment in the fungus gardens of attine ants. In the present study, the production of proteolytic enzymes was observed in 32% and 16% of the yeasts isolated from A. texana and Acromyrmex, respectively. The community of yeasts and the proteolytic bacteria associated with nests [29,30] could contribute to the degradation of proteins present in the plant material and generate available nitrogen, which together with the nitrogen incorporated by the action of nitrogen-fixing bacteria [36] would support the development of the mutualistic fungus, ants and other microbes.
Xylanolytic enzymes were exhibited by 59% of the yeasts isolated from A. texana gardens. Xylanase is the second most abundant enzyme exhibited by the mutualistic fungus [3]. Both ants and the cultivar may have mutual development supported by xylose [3,7], the hydrolysis product of xylan. Xylan-degrading yeasts may contribute to the degradation of hemicellulose present in the plant material used by the fungal cultivar. Moreover, all yeast isolates screened in the present study were able to assimilate xylose (Table 1 and Table 2), suggesting that this carbon source would also be important in the nutrition of yeasts.
It seems that the degradation of polymers present in the fungus gardens is the result of a complementary action of enzymes exhibited by the mutualistic fungus, bacteria and also yeasts that are found in the fungus garden. Such microbial consortium would generate readily available nutrients that may help to sustain the homeostasis of the attine ant-microbe symbiosis.

4. Conclusion

Our results demonstrated that yeasts found in attine gardens exhibit hydrolytic enzymes capable of breaking down the plant polysaccharides found in the substrate used to culture the mutualistic fungus. In addition, we showed that yeasts are able to grow on most of the oligosaccharides derived from the digestion of the plant polysaccharides. Thus, the observed enzymatic capacity of yeasts would contribute to the ant nest by generating readily available nutrients for their own growth or for the growth of other organisms involved in the symbiosis. Consequently, by doing so, yeasts may contribute to the detoxification of compounds such as galacturonic acid that is potentially harmful to the ants and not assimilated by the mutualistic fungus.

Acknowledgments

We would like to thank FAPESP (Fundação de Amparo à Pesquisa do Estado de São Paulo) and CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico) for financial support. We also thank CAPES (Coordenação de Aperfeiçoamento de Pessoal de Ensino Superior) and TWAS/CNPq for providing a graduate fellowship to T.D. Mendes and I. Dayo-Owoyemi, respectively. The authors also thank M. Bacci Jr., A. Ortiz and U.G. Mueller for collecting the ant nests from South Brazil and kindly provide the fungus garden material used in this study. We thank four anonymous reviewers and the guest editor for helpful comments on this manuscript.

References

  1. Hölldobler, E.; Wilson, E.O. Journey to the Ants: A Story of Scientific Exploration, 3rd ed.; Harvard University Press: Cambridge, London, UK, 1995. [Google Scholar]
  2. Currie, C.R. A community of ants, fungi and bacteria: A multilateral approach to study symbiosis. Annu. Rev. Microbiol. 2001, 55, 357–380. [Google Scholar] [CrossRef]
  3. Siqueira, C.G.; Bacci, M., Jr.; Pagnocca, F.C.; Bueno, O.C.; Hebling, M.J.A. Metabolism of plant polysaccharides by Leucoagaricus gongylophorus, the symbiotic fungus of the leaf-cutting ant Atta sexdens L. Appl. Environ. Microbiol. 1998, 64, 4820–4822. [Google Scholar]
  4. Silva, A.; Bacci, M., Jr.; Pagnocca, F.C.; Bueno, O.C.; Hebling, M.J.A. Production of polysaccharidases in different carbon sources by Leucoagaricus gongylophorus Möller (Singer), the symbiotic fungus of the leaf-cutting ant Atta sexdens Linnaeus. Curr. Microbiol. 2006, 53, 68–71. [Google Scholar]
  5. Schiott, M.; Rogowska-Wrzesinka, A.; Roepstorff, P.; Boomsma, J.J. Leaf-cutting ant fungi produce cell wall degrading pectinase complexes reminiscent of phytopathogenic fungi. BMC Biol. 2010, 8, 1–12. [Google Scholar] [CrossRef]
  6. Kooij, P.W.; Schiott, M.; Boomsma, J.J.; de Fine Licht, H.H. Rapid shifts in Atta cephalotes fungus-garden enzyme activity after a change in fungal substrate (Attini, Formicidae). Insectes Soc. 2011, 58, 145–151. [Google Scholar] [CrossRef]
  7. Silva, A.; Bacci, M., Jr.; Siqueira, G.C.; Bueno, O.C.; Pagnocca, F.C.; Hebling, M.J.A. Survival of Atta sexdens workers on different food sources. J. Insect Physiol. 2003, 49, 307–313. [Google Scholar] [CrossRef]
  8. Weber, N.A. Gardening Ants: The Attines, Memoirs of American Philosophical Society; American Philosophical Society: Philadelphia, PA, USA, 1972; Volume 92, pp. 1–146. [Google Scholar]
  9. Currie, C.R.; Stuart, A.E. Weeding and grooming of pathogens in agriculture by ants. Proc. R. Soc. Lond. B Biol. Sci. 2001, 268, 1033–1039. [Google Scholar] [CrossRef]
  10. Beattie, A.J.; Turnbull, C.L.; Hough, T.; Knox, R.B. Antibiotic production: A possible function for the metapleural glands of ants (Hymenoptera: Formicidae). Ann. Entomol. Soc. Am. 1986, 79, 448–450. [Google Scholar]
  11. Nascimento, R.R.; Schoeters, E.; Morgan, E.D.; Billen, J.; Stradling, D.J. Chemistry of metapleural gland secretions of three attine ants, Atta sexdens rubropilosa, Atta cephalotes, and Acromyrmex octospinosus (Hymenoptera: Formicidae). J. Chem. Ecol. 1996, 22, 987–1000. [Google Scholar] [CrossRef]
  12. Poulsen, M.; Hughes, W.O.H.; Boomsma, J.J. Differential resistance and the importance of antibiotic production in Acromyrmex echinatior leaf-cutting ant castes towards the entomopathogenic fungus Aspergillus nomius. Insectes Soc. 2006, 53, 349–355. [Google Scholar] [CrossRef]
  13. Marsaro, A.L., Jr.; Della Lucia, T.M.C.; Barbosa, L.C.A.; Mafia, L.A.; Morandi, M.A.B. Efeito de secreções da glândula mandibular de Atta sexdens rubropilosa Forel (Hymenoptera: Formicidae) sobre a germinação de conídios de Botrytis cinerea Pers. Fr. Neotrop. Entomol. 2001, 30, 403–406. [Google Scholar] [CrossRef]
  14. Rodrigues, A.; Carletti, C.D.; Bueno, O.C.; Pagnocca, F.C. Leaf-cutting ant faecal fluid and mandibular gland secretion: Effects on microfungi spore germination. Braz. J. Microbiol. 2008, 39, 64–67. [Google Scholar] [CrossRef]
  15. Currie, C.R.; Scott, J.A.; Summerbell, R.C.; Malloch, D. Fungus-growing ants use antibiotic-producing bacteria to control garden parasites. Nature 1999, 398, 701–704. [Google Scholar]
  16. Currie, C.R.; Scott, J.A.; Summerbell, R.C.; Malloch, D. Corrigendum: Fungus-growing ants use antibiotic-producing bacteria to control garden parasites. Nature 2003, 423, 461. [Google Scholar]
  17. Sen, R.; Ishak, H.D.; Estrada, D.; Dowd, S.E.; Hong, E.; Mueller, U.G. Generalized antifungal activity and 454-screening of Pseudonocardia and Amycolatopsis bacteria in nests of fungus-growing ants. Proc. Natl. Acad. Sci. USA 2009, 106, 17805–17810. [Google Scholar]
  18. Mueller, U.G.; Dash, D.; Rabeling, C.; Rodrigues, A. Coevolution between attine ants and actinomycete bacteria: A reevaluation. Evolution 2008, 62, 2894–2912. [Google Scholar] [CrossRef]
  19. Zucchi, T.D.; Guidolin, A.S.; Cônsoli, F.L. Isolation and characterization of actinobacteria ectosymbionts from Acromyrmex subterraneus brunneus (Hymenoptera, Formicidae). Microbiol. Res. 2011, 166, 68–76. [Google Scholar] [CrossRef]
  20. Reynolds, H.T.; Currie, C.R. Pathogenicity of Escovopsis weberi: The parasite of the attine ant-microbe symbiosis directly consumes the ant-cultivated fungus. Mycologia 2004, 96, 955–959. [Google Scholar] [CrossRef]
  21. Fisher, P.J.; Stradling, D.J.; Sutton, B.C.; Petrini, L.E. Microfungi in the fungus gardens of the leaf-cutting ant Atta cephalotes: A preliminary study. Mycol. Res. 1996, 100, 541–546. [Google Scholar] [CrossRef]
  22. Craven, S.E.; Dix, M.W.; Michaelis, G.E. Attine fungus garden contain yeasts. Science 1970, 169, 184–189. [Google Scholar]
  23. Angelis, C.; Serzedello, A.; de Angelis, D.F. Yeasts found in gardens of Atta sexdens rubropilosa and Atta laevigata. Naturalia 1983, 8, 149–151. [Google Scholar]
  24. Carreiro, S.C.; Pagnocca, F.C.; Bueno, O.C.; Bacci, M., Jr.; Hebling, M.J. A.; da Silva, O.A. Yeasts associated with nests of the leaf-cutting ant Atta sexdens rubropilosa Forel, 1908. Antonie van Leeuwenhoek 1997, 71, 243–248. [Google Scholar] [CrossRef]
  25. Pagnocca, F.C.; Rodrigues, A.; Nagamoto, N.S.; Bacci, M., Jr. Yeasts and filamentous fungi carried by the gynes of leaf-cutting ants. Antonie van Leeuwenhoek 2008, 94, 517–526. [Google Scholar] [CrossRef]
  26. Guedes, A.F.A.; Attili-Angelis, D.; Pagnocca, F.C. Selective isolation of dematiaceous fungi from the workers of Atta laevigata (Formicidae: Attini). Folia Microbiol. 2011. [Google Scholar]
  27. Little, A.E.F.; Currie, C.R. Symbiotic complexity: Discovery of a fifth symbiont in the attine ant-microbe symbiosis. Biol. Lett. 2007, 3, 501–504. [Google Scholar] [CrossRef]
  28. Rodrigues, A.; Cable, R.N.; Mueller, U.G.; Bacci, M., Jr.; Pagnocca, F.C. Antagonistic interactions between gardens yeasts and microfungal garden pathogens of leaf-cutting ants. Antonie van Leeuwenhoek 2009, 96, 331–342. [Google Scholar] [CrossRef]
  29. Ribeiro, S.B. Caracterização de espécies bacterianas encontradas em ninhos de Atta sexdens L. e isolamento de Streptomyces de formigas da tribo Attini. Ph.D. Thesis, Instituto de Biociências, Universidade Estadual Paulista, Rio Claro, Brazil, 2000. [Google Scholar]
  30. Bacci, M., Jr.; Ribeiro, S.B.; Casarotto, M.E.F.; Pagnocca, F.C. Biopolymer-degrading bacteria from nests of the leaf-cutting ant Atta sexdens rubropilosa. Braz. J. Med. Biol. Res. 1995, 28, 79–82. [Google Scholar]
  31. Rodrigues, A.; Bacci, M., Jr.; Mueller, U.G.; Ortiz, A.; Pagnocca, F.C. Microfungal “weeds” in the leafcutter ant symbiosis. Microb. Ecol. 2008, 56, 604–614. [Google Scholar] [CrossRef]
  32. Carreiro, S.C.; Pagnocca, F.C.; Bacci, M., Jr.; Lachance, M.A.; Bueno, O.C.; Hebling, M.J.A.; Ruivo, C.C.; Rosa, C.A. Simpodiomyces attinorum sp. nov., a yeast species associated with nests of the leaf-cutting ant Atta sexdens. Int. J. Syst. Evol. Microbiol. 2004, 54, 1891–1894. [Google Scholar]
  33. Middelhoven, W.J.; Fonseca, A.; Carreiro, S.C.; Pagnocca, F.C.; Bueno, O.C. Cryptococcus haglerorum sp. nov., an anamorphic basidiomycetous yeast isolated from nests of the leaf-cutting ant Atta sexdens. Antonie van Leeuwenhoek 2003, 83, 167–174. [Google Scholar] [CrossRef]
  34. Pagnocca, F.C.; Legaspe, M.F.C.; Rodrigues, A.; Ruivo, C.C.C.; Nagamoto, N.S.; Bacci, M., Jr.; Forti, L.C. Yeast isolated from a fungus-growing ant nest including the description of Trichosporon chiarellii sp. nov., an anamorphic basidiomycetous yeast. Int. J. Syst. Evol. Microbiol. 2010, 60, 1454–1459. [Google Scholar] [CrossRef]
  35. Suen, G.; Scott, J.J.; Aylward, F.O.; Adams, S.M.; Tringe, S.G.; Pinto-Tomás, A.A.; Foster, C.E.; Pauly, M.; Weimer, P.J.; Barry, K.W.; et al. An insect herbivore microbiome with high plant biomass-degrading capacity. PloS Genet. 2011, 6, 1–14. [Google Scholar]
  36. Pinto-Tomás, A.A.; Anderson, M.A.; Suen, G.; Stevenson, D.M.; Chu, F.S.T.; Cleland, W.W.; Weimer, P.J.; Currie, C.R. Symbiotic nitrogen fixation in the fungus gardens of leaf-cutter ants. Science 2009, 326, 1120–1123. [Google Scholar]
  37. de Almeida, J.M.G.C.F. Yeast community survey in the Tagus estuary. FEMS Microbiol. Lett. 2005, 52, 295–303. [Google Scholar]
  38. Sampaio, J.P.; Gadanho, M.; Santos, S.; Duarte, F.L.; Pais, C.; Fonseca, A.; Fell, J.W. Polyphasic taxonomy of basidiomycetous yeasts genus Rhodosporidium: Rhodosporidium kratochvilovae and related anamorphic species. Int. J. Syst. Evol. Microbiol. 2001, 51, 687–697. [Google Scholar]
  39. Kurtzman, C.P.; Robnett, C.J. Identification and phylogeny of ascomycetous yeasts from analysis of nuclear large subunit (26S) ribosomal DNA partial sequences. Antonie van Leeuwenhoek 1998, 73, 331–371. [Google Scholar] [CrossRef]
  40. Hall, T.A. BioEdit: A user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp. Ser. 1999, 41, 95–98. [Google Scholar]
  41. Altschul, S.F.; Madden, T.L.; Schaffer, A.A.; Zhang, J.; Zhang, Z.; Miller, W.; Lipman, D. Gapped BLAST and PSI-BLAST: A new generation of protein database search programs. Nucleic Acids Res. 1997, 25, 3389–3402. [Google Scholar]
  42. Looder, J. The Yeast,a Taxonomic Study, 2nd ed.; North-Holland Pub. Co.: Amsterdam, The Netherlands, 1970. [Google Scholar]
  43. Maijala, P.; Fagerstedt, K.V.; Raudaskoski, M. Detection of extracellular cellulolytic and proteolytic activity in ectomycorrhyzal fungi and Hetererobasidion annosum (Fr.) Bref. New Phytol. 1991, 117, 643–648. [Google Scholar] [CrossRef]
  44. Mckay, A.M. A plate assay method for the detection of fungal polygalacturonase secretion. FEMS Microbiol. Lett. 1988, 56, 355–358. [Google Scholar] [CrossRef]
  45. Williams, A.G. Staining reactions for the detection of hemicellulose-degrading bacteria. FEMS Microbiol. Lett. 1983, 20, 253–358. [Google Scholar] [CrossRef]
  46. Carreiro, S.C.; Pagnocca, F.C.; Bacci, M., Jr.; Bueno, O.C.; Hebling, M.J.A.; Middelhoven, W.J. Occurrence of killer yeasts in leaf-cutting ant nests. Folia Microbiol. 2002, 47, 259–262. [Google Scholar] [CrossRef]
  47. Inacio, J.; Pereira, P.; de Carvalho, M.; Fonseca, A.; Amaral-Collaço, M.T.; Spencer-Martins, I. Estimation and diversity of phylloplane mycobiota on selected plants in a Mediterranean-type ecosystem in Portugal. Microb. Ecol. 2002, 44, 344–353. [Google Scholar] [CrossRef]
  48. Glushakova, A.M.; Chernov, I.Y. Seasonal dynamics of the structure of epiphytic yeasts communities. Microbiology 2010, 79, 830–839. [Google Scholar] [CrossRef]
  49. Spencer, J.F.T.; Spencer, D.M. Yeasts in Natural and Artificial Habitats; Springer: Berlin, Germany, 1997. [Google Scholar]
  50. Sosa-Calvo, J.; Schultz, T.R. Three remarkable new fungus-growing ant species of the genus Myrmicocrypta (Hymenoptera: Formicidae), with a reassessment of the characters that define the genus and its position within the Attini. Ann. Entomol. Soc. Am. 2010, 103, 181–195. [Google Scholar] [CrossRef]
  51. Phaff, H.J.; Starmer, W.T. Yeasts associated with plants, insects and soil. In The Yeasts, 2nd ed.; Rose, A.H., Harrison, J.S., Eds.; Academic Press: London, UK, 1987; pp. 123–180. [Google Scholar]
  52. Papa, P.J.; Papa, F. Inhibition des bactéries dans les nids d’Acromyrmex octospinosus Reich. Bull. Soc. Pathol. Exot. 1982, 75, 415–425. [Google Scholar]
  53. Sakay, T.; Sakamoto, T.; Hallaert, J.; Vandamme, E.J. Pectin, pectinase, and protopectinase: Production, properties, and applications. Adv. Appl. Microbiol. 1993, 39, 213–293. [Google Scholar] [CrossRef]
  54. Carreiro, S.C. Pesquisa de fator killer e análise de degradação de polissacarídeos vegetais por leveduras associadas aos ninhos de Atta sexdens. Ph.D. Thesis, Instituto de Biociências de Rio Claro, Universidade Estadual Paulista, Rio Claro, Brazil, 1999. [Google Scholar]
  55. Stolp, H. Microbial Ecology: Organisms, Habitats, Activities, 1st ed.; Cambridge University Press: London, UK, 1988. [Google Scholar]
  56. Martin, M.M.; Weber, N.A. The celullose-utilizing capability of the fungus cultured by the attine ant Atta colombica tonsipes. Ann. Entomol. Soc. Am. 1969, 62, 1386–1387. [Google Scholar]
  57. Bacci, M., Jr.; Anversa, M.M.; Pagnocca, F.C. Cellulose degradation by Leucocoprinus gongylophorus, the fungus cultured by the leaf-cutting ant Atta sexdens rubropilosa. Antonie van Leeuwenhoek 1995, 67, 385–386. [Google Scholar] [CrossRef]
  58. Abril, A.B.; Bucher, E.H. Evidence that the fungus cultured by leaf-cutting ants does not metabolize cellulose. Ecol. Lett. 2002, 5, 325–328. [Google Scholar] [CrossRef]
  59. Silva, A.; Bacci, M., Jr.; Pagnocca, F.C.; Bueno, O.C.; Hebling, M.J.A. Starch metabolism in Leucoagaricus gongylophorus, the symbiotic fungus of leaf-cutting ants. Microbiol. Res. 2005, 161, 299–303. [Google Scholar]
  60. Kurtzman, C.P.; Fell, J.W.; Boekhout, T. The Yeasts: A Taxonomic Study, 5th ed.; Elsevier: San Diego, CA, USA, 2011. [Google Scholar]
  61. Abril, A.B.; Bucher, E.H. Nutritional sources of the fungus cultured by leaf-cutting ants. Appl. Soil Ecol. 2003, 26, 243–247. [Google Scholar]

Supplemental Material

Insects 03 00228 g002 1024
Figure S1. Polymer hydrolysis assays. Figure shows zones of hydrolysis of the tested polymers: (A) Amylase, produced by Pseudozyma sp. (ATT068); (B) CMCellulase, produced by Trichosporon montevideense (88c1); (C) Pectinase, produced by Pseudozyma sp. (ATT068); (D) Xylanase, produced by Cryptococcus cf. cellulolyticus (ATT064) and (E) Protease, produced by Pseudozyma sp. (ATT068).
Figure S1. Polymer hydrolysis assays. Figure shows zones of hydrolysis of the tested polymers: (A) Amylase, produced by Pseudozyma sp. (ATT068); (B) CMCellulase, produced by Trichosporon montevideense (88c1); (C) Pectinase, produced by Pseudozyma sp. (ATT068); (D) Xylanase, produced by Cryptococcus cf. cellulolyticus (ATT064) and (E) Protease, produced by Pseudozyma sp. (ATT068).
Insects 03 00228 g002
Table
Table S1. Yeast and yeast-like strains examined.
Table S1. Yeast and yeast-like strains examined.
Strain CodeYeast SpeciesNest IDAnt SpeciesCity/State/Country 1Nest Location 2
8aGalactomyces candidusAOMB100904-04Acromyrmex heyeriSentinela do Sul/RS/BrazilS 30º37'09''; W 51º33'18''
10aMeyerozyma guilliermondiiAOMB120904-05Acromyrmex subterraneusSantana da Boa Vista/RS/BrazilS 31º19'35''; W 52º44'40''
15aGalactomyces candidus
16a1Meyerozyma guilliermondiiAOMB130904-02Acromyrmex coronatusVacaria/RS/BrazilS 28º27'51''; W 50º53'07''
16a2Meyerozyma guilliermondii
24aGalactomyces candidusAOMB040904-02Acromyrmex coronatusPiraquara/PR/BrazilS 25º25'50''; W 49º04'56''
27aGalactomyces candidusAOMB110904-04Acromyrmex lundiChuvisca/RS/BrazilS 30º50'10''; W 51º55'10''
27a1Galactomyces candidus
27b1Trichosporon chiarellii
27c1Trichosporon chiarellii
27c2’Cryptococcus laurentii
28b2Galactomyces candidusAOMB110904-05Acromyrmex lundiChuvisca/RS/BrazilS 30º50'10''; W 51º55'10''
30bTrichosporon chiarelliiAOMB110904-03Acromyrmex heyeriChuvisca/RS/BrazilS 30º50'10''; W 51º55'10''
30cTrichosporon chiarellii
31aGalactomyces candidusAOMB120904-09Acromyrmex sp.Santana da Boa Vista/RS/BrazilS 30º56'40''; W 53º05'10''
31bTrichosporon multisporum
31b2Trichosporon montevideense
31cGalactomyces candidus
32aTrichosporon multisporumAOMB120904-03Acromyrmex ambiguusSantana da Boa Vista/RS/BrazilS 31º19'35''; W 52º44'40''
32cTrichosporon multisporum
39aMeyerozyma caribbicaAOMB140904-03Acromyrmex discigerBlumenau/SC/BrazilS 26º54'04''; W 49º10'51''
39bGalactomyces candidus
56bGalactomyces candidusAOMB140904-05Acromyrmex laticepsBlumenau/SC/BrazilS 26º54'04''; W 49º10'51''
59aGalactomyces candidusAOMB100904-07Acromyrmex heyeriSentinela do Sul/RS/BrazilS 30º37'09''; W 51º33'18''
59bGalactomyces candidus
63aTrichosporon chiarelliiAOMB110904-11Acromyrmex lundiChuvisca/RS/BrazilS 30º50'10''; W 51º55'10''
63cTrichosporon chiarellii
77bTrichosporon montevideenseAOMB110904-20Acromyrmex laticepsChuvisca/RS/BrazilS 30º50'10''; W 51º55'10''
78aMeyerozyma guilliermondiiAOMB130904-10Acromyrmex laticepsLages/SC/Brazil-
78bMeyerozyma guilliermondii
78b2Meyerozyma guilliermondii
79bGalactomyces candidusAOMB120904-01Acromyrmex ambiguusSantana da Boa Vista/RS/BrazilS 31º19'35''; W 52º44'40''
87aGalactomyces candidusAOMB110904-10Acromyrmex laticepsChuvisca/RS/BrazilS 30º50'10''; W 51º55'10''
88a1Trichosporon montevideenseAOMB060904-03Acromyrmex ambiguusNova Petrópolis/RS/BrazilS 29º23'18''; W 50º54'40''
88a2Trichosporon montevideense
88bTrichosporon montevideense
88b2Galactomyces candidus
88c1Trichosporon montevideense
ATT001Candida membranifaciensUGM051218-02Atta texanaBastrop County/TX/USAN 30°05'49''; W 97°13'29''
ATT002Candida membranifaciens
ATT004Candida membranifaciens
ATT003Candida melibiosica
ATT175unidentified yeast-like fungus
ATT176Cryptococcus sp. 4
ATT177Rhodotorula nothofagi
ATT178Cryptococcus sp.
ATT203unidentified yeast-like fungus
ATT204Cryptococcus podzolicus
ATT205Kodamaea ohmeri
ATT252Rhodotorula mucilaginosa
ATT253Cryptococcus laurentii
ATT254Sporidiobolus ruineniae
ATT256Sporidiobolus ruineniae
ATT255Sporisorium penniseti (yeast-like)
ATT257Sporisorium penniseti (yeast-like)
ATT258Rhodotorula lactosa
ATT259Cryptococcus flavus
ATT064Cryptococcus cf. cellulolyticusUGM060121-02Atta texanaAustin/TX/USAN 30°13'56.40"; W 97°39'10.80"
ATT067Cryptococcus cf. cellulolyticus
ATT068Pseudozyma sp.
ATT069Cryptococcus magnus
ATT070Farysizyma sp. 3
ATT121Cryptococcus magnus
ATT122Cryptococcus luteolus
ATT123Cryptococcus sp. 3
ATT147Rhodotorula sp. HB 1211
ATT148Cryptococcus magnus
ATT260Cryptococcus flavus
ATT262Aureobasidium pullulans
ATT263Aureobasidium pullulans
ATT269Aureobasidium pullulans
ATT073Cryptococcus laurentiiUGM060121-01Atta texanaAustin/TX/USAN 30°13'58.38"; W 97°39'06.06"
ATT075Cryptococcus laurentii
ATT074Bullera sinensis
ATT078Bulleromyces albus
ATT079Cryptococcus sp. 1
ATT080Cryptococcus sp. 2
ATT082Hannaella kunmingensis 3
ATT120Cryptococcus flavescens
ATT264Sympodiomycopsis paphiopedili
ATT268Cryptococcus flavus
ATT271Sympodiomycopsis paphiopedili
1 PR: Paraná; RS: Rio Grande do Sul; SC: Santa Catarina; TX: Texas; 2 GPS data unit: dd mm ss and dd mm ss.ss; 3 These strains were previously identified in Rodrigues et al. [28] as Rhodotorula cf. taiwaniana and Cryptococcus cf. luteolus and now are confirmed to belong to Farysizyma sp. and H. kunmingensis, respectively.
Table
Table S2. Enzymatic activity and assimilation of compounds by yeast and yeast-like fungi examined in this study.
Table S2. Enzymatic activity and assimilation of compounds by yeast and yeast-like fungi examined in this study.
Strain CodeYeast SpeciesHydrolytic Enzymes 1 Assimilation 2
AMCELPXYLPRTMCAX
27c2’Cryptococcus laurentii 4---ND 3+ ++++
8aGalactomyces candidus 4-++ND- --++
15aGalactomyces candidus 4-++ND+ --++
24aGalactomyces candidus-++ND- --++
27aGalactomyces candidus-++ND- --++
27a1Galactomyces candidus-++ND- --++
28b2Galactomyces candidus-++ND- --++
31aGalactomyces candidus-++ND- --++
31cGalactomyces candidus-++ND- --++
39bGalactomyces candidus-++ND- --++
56bGalactomyces candidus-+-ND- --++
59aGalactomyces candidus-++ND- --++
59bGalactomyces candidus-++ND- --++
79bGalactomyces candidus-++ND+ --++
87aGalactomyces candidus-+-ND- --++
88b2Galactomyces candidus-+-ND- ---+
39aMeyerozyma caribbica 4-+-ND- ++++
10aMeyerozyma guilliermondii-++ND- ++++
16a1Meyerozyma guilliermondii 4-++ND- ++-+
16a2Meyerozyma guilliermondii-++ND- ++-+
78aMeyerozyma guilliermondii-++ND+ ++++
78bMeyerozyma guilliermondii-++ND- ++++
78b2Meyerozyma guilliermondii-++ND- ++-+
27b1Trichosporon chiarellii--+ND- ++++
27c1Trichosporon chiarellii-+-ND- ++++
30bTrichosporon chiarellii--+ND- ++++
30cTrichosporon chiarellii 4--+ND- ++++
63aTrichosporon chiarellii-++ND+ ++++
63cTrichosporon chiarellii-+-ND- ++++
31b2Trichosporon montevideense-++ND+ ++++
77bTrichosporon montevideense+++ND- ++++
88a1Trichosporon montevideense-+-ND- ++-+
88a2Trichosporon montevideense-+-ND- ++-+
88bTrichosporon montevideense-+-ND- ++-+
88c1Trichosporon montevideense 4-+-ND- ++++
31bTrichosporon multisporum 4+++ND- ++++
32aTrichosporon multisporum+++ND- ++-+
32cTrichosporon multisporum-++ND- ++++
ATT262Aureobasidium pullulans-+-+- ++++
ATT263Aureobasidium pullulans-+-++ ++++
ATT269Aureobasidium pullulans-+-++ ++-+
ATT003Candida melibiosica----- ++-+
ATT001Candida membranifaciens----- ++-+
ATT002Candida membranifaciens----- ++-+
ATT004Candida membranifaciens----- ++-+
ATT074Bullera sinensis++-+- ++++
ATT078Bulleromyces albus-+-+- ++++
ATT064Cryptococcus cf. cellulolyticus-+-+- ++++
ATT067Cryptococcus cf. cellulolyticus-+-+- ++++
ATT259Cryptococcus flavus++-++ ++++
ATT260Cryptococcus flavus++-++ ++++
ATT268Cryptococcus flavus++++- ++++
ATT120Cryptococcus flavescens---+- ++++
ATT253Cryptococcus laurentii+-++- ++++
ATT073Cryptococcus laurentii++-+- ++++
ATT075Cryptococcus laurentii++--- ++++
ATT122Cryptococcus luteolus----+ ++++
ATT069Cryptococcus magnus-+-++ ++-+
ATT148Cryptococcus magnus-+--+ ++-+
ATT121Cryptococcus magnus-+-+- ++-+
ATT204Cryptococcus podzolicus+--+- ++++
ATT178Cryptococcus sp.---+- ++++
ATT079Cryptococcus sp. 1-+-+- ++-+
ATT080Cryptococcus sp. 2----- ++-+
ATT123Cryptococcus sp. 3----- ++++
ATT176Cryptococcus sp. 4+-++- ++++
ATT070Farysizyma sp.---++ ++++
ATT082Hannaella kunmingensis----- ++++
ATT205Kodamaea ohmeri----- ++-+
ATT068Pseudozyma sp.+++++ ++++
ATT258Rhodotorula lactose----- ++++
ATT252Rhodotorula mucilaginosa--+-- ++++
ATT177Rhodotorula nothofagi+-++- ++-+
ATT147Rhodotorula sp. HB 1211----- ++++
ATT254Sporidiobolus ruineniae--+-- ++-+
ATT256Sporidiobolus ruineniae--+-- ++-+
ATT255Sporisorium penniseti (yeast-like)--+++ ++++
ATT257Sporisorium penniseti (yeast-like)----+ ++++
ATT264Sympodiomycopsis paphiopedili-+-+- ++-+
ATT271Sympodiomycopsis paphiopedili---++ ++-+
ATT175unidentified yeast-like fungus-+-++ ++++
ATT203unidentified yeast-like fungus----+ ++++
1 AM: Amylase; CEL: CMCellulase; P: pectinase; XYL: xylanase; PRT: protease; 2 M: maltose; C: cellobiose; A: galacturonic acid; X: xylose; 3 ND: xylanase was not determined for yeasts isolated from Acromyrmex spp.; 4 Strains selected as representative for DNA sequencing.

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Mendes, T.D.; Rodrigues, A.; Dayo-Owoyemi, I.; Marson, F.A.L.; Pagnocca, F.C. Generation of Nutrients and Detoxification: Possible Roles of Yeasts in Leaf-Cutting Ant Nests. Insects 2012, 3, 228-245. https://doi.org/10.3390/insects3010228

AMA Style

Mendes TD, Rodrigues A, Dayo-Owoyemi I, Marson FAL, Pagnocca FC. Generation of Nutrients and Detoxification: Possible Roles of Yeasts in Leaf-Cutting Ant Nests. Insects. 2012; 3(1):228-245. https://doi.org/10.3390/insects3010228

Chicago/Turabian Style

Mendes, Thais D., André Rodrigues, Ifeloju Dayo-Owoyemi, Fernando A. L. Marson, and Fernando C. Pagnocca. 2012. "Generation of Nutrients and Detoxification: Possible Roles of Yeasts in Leaf-Cutting Ant Nests" Insects 3, no. 1: 228-245. https://doi.org/10.3390/insects3010228

APA Style

Mendes, T. D., Rodrigues, A., Dayo-Owoyemi, I., Marson, F. A. L., & Pagnocca, F. C. (2012). Generation of Nutrients and Detoxification: Possible Roles of Yeasts in Leaf-Cutting Ant Nests. Insects, 3(1), 228-245. https://doi.org/10.3390/insects3010228

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