Antifungal Streptomyces spp., Plausible Partners for Brood-Caring of the Dung Beetle Copris tripartitus
Abstract
:1. Introduction
2. Materials and Methods
2.1. Sample Collection
2.2. Microbial Isolation and Identification
2.3. Antifungal Screening by the Dual Culture Assay
2.4. Extraction of Nucleic Acids from Samples
2.5. Nested PCR Amplification
2.6. DGGE Analysis
2.7. Identification of DGGE Bands
2.8. HPLC analysis of Culture Extracts
2.9. Culture and Extraction of Strain AT67
2.10. Separation and Bioassay of Bioactive Fraction
3. Results
3.1. Microbial Isolation from the Dung Beetle and Brood Balls
3.2. Antifungal Activities of Isolates Derived from the Dung Beetle
3.3. Identification and Phylogenetic Analysis of Antifungal Isolates
3.4. Association between the Dung Beetle and Antifungal Isolates
3.5. Chemical Profile of 16 Antifungal Strains
3.6. Antifungal Metabolite Produced by Strain AT67
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Bang, H.-S.; Crespo, C.H.; Na, Y.-E.; Han, M.-S.; Lee, J.-H. Reproductive development and seasonal activity of two Korean native Coprini species (Coleoptera: Scarabaeidae). J. Asia-Pac. Èntomol. 2008, 11, 195–199. [Google Scholar] [CrossRef]
- Bang, H.S.; Lee, J.-H.; Kwon, O.S.; Na, Y.E.; Jang, Y.S.; Kim, W.H. Effects of paracoprid dung beetles (Coleoptera: Scarabaeidae) on the growth of pasture herbage and on the underlying soil. Appl. Soil Ecol. 2005, 29, 165–171. [Google Scholar] [CrossRef]
- Hanski, I.; Cambefort, Y. Dung Beetle Ecology; Princeton University Press: Princeton, NJ, USA, 1991; pp. 481–486. [Google Scholar]
- Scholtz, C.H.; Davis, A.L.V.; Kryger, U. Evolutionary Biology and Conservation of Dung Beetles; Pensoft Sofia-Moscow: Moscow, Russia, 2009. [Google Scholar]
- Bang, H.S.; Huerta, C.; Kim, J.I.; Goo, T.W. Studies on the Ecology of Oviposition of Copris tripartitus Waterhouse and Copris ochus (Motschulsky) (Coleoptera: Scarabaeidae). Korean J. Entomol. 2001, 31, 237–242. [Google Scholar]
- Tyndale-Biscoe, M. Adaptive significance of brood care of Copris diversus Waterhouse (Coleoptera: Scarabaeidae). Bull. Èntomol. Res. 1984, 74, 453–461. [Google Scholar] [CrossRef]
- Halffter, G.; Huerta, C.; Lopez-Portillo, J. Parental care and offspring survival inCopris incertusSay, a sub-social beetle. Anim. Behav. 1996, 52, 133–139. [Google Scholar] [CrossRef] [Green Version]
- Halffter, G.; Edmonds, W.D. The nesting behavior of dung beetles (Scarabaeinae). In An Ecological and Evolutive Approach. The Nesting Behavior of Dung Beetles (Scarabaeinae); Instituto de Ecologia: Ciudad de Mexico, Mexico, 1982. [Google Scholar]
- Byrne, M.J.; Watkins, B.; Bouwer, G. Do dung beetle larvae need microbial symbionts from their parents to feed on dung? Ecol. Entomol. 2013, 38, 250–257. [Google Scholar] [CrossRef]
- Florez, L.V.; Biedermann, P.; Engl, T.; Kaltenpoth, M. Defensive symbioses of animals with prokaryotic and eukaryotic microorganisms. Nat. Prod. Rep. 2015, 32, 904–936. [Google Scholar] [CrossRef] [Green Version]
- Currie, C.R.; Scott, J.; Summerbell, R.C.; Malloch, D. Fungus-growing ants use antibiotic-producing bacteria to control garden parasites. Nature 1999, 398, 701–704. [Google Scholar] [CrossRef]
- Kaltenpoth, M.; Goettler, W.; Dale, C.; Stubblefield, J.W.; Herzner, G.; Roeser-Mueller, K.; Strohm, E. ‘Candidatus Streptomyces philanthi’, an endosymbiotic streptomycete in the antennae of Philanthus digger wasps. Int. J. Syst. Evol. Microbiol. 2006, 56, 1403–1411. [Google Scholar] [CrossRef] [PubMed]
- Scott, J.; Oh, D.-C.; Yuceer, M.C.; Klepzig, K.D.; Clardy, J.; Currie, C.R. Bacterial Protection of Beetle-Fungus Mutualism. Science 2008, 322, 63. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Vasquez, A.; Forsgren, E.; Fries, I.; Paxton, R.J.; Flaberg, E.; Szekely, L.; Olofsson, T.C. Symbionts as Major Modulators of Insect Health: Lactic Acid Bacteria and Honeybees. PLoS ONE 2012, 7, e33188. [Google Scholar] [CrossRef]
- Oh, D.-C.; Poulsen, M.; Currie, C.R.; Clardy, J. Dentigerumycin: A bacterial mediator of an ant-fungus symbiosis. Nat. Chem. Biol. 2009, 5, 391–393. [Google Scholar] [CrossRef] [PubMed]
- Kroiss, J.; Kaltenpoth, M.; Schneider, B.; Schwinger, M.-G.; Hertweck, C.; Maddula, R.K.; Strohm, E.; Svatos, A. Symbiotic streptomycetes provide antibiotic combination prophylaxis for wasp offspring. Nat. Chem. Biol. 2010, 6, 261–263. [Google Scholar] [CrossRef]
- Van Arnam, E.B.; Currie, C.R.; Clardy, J. Defense contracts: Molecular protection in insect-microbe symbioses. Chem. Soc. Rev. 2017, 47, 1638–1651. [Google Scholar] [CrossRef]
- Estes, A.M.; Hearn, D.J.; Snell-Rood, E.C.; Feindler, M.; Feeser, K.; Abebe, T.; Hotopp, J.C.D.; Moczek, A.P. Brood Ball-Mediated Transmission of Microbiome Members in the Dung Beetle, Onthophagus taurus (Coleoptera: Scarabaeidae). PLoS ONE 2013, 8, e79061. [Google Scholar] [CrossRef]
- Kim, S.-H.; Ko, H.; Bang, H.-S.; Park, S.-H.; Kim, D.-G.; Kwon, H.C.; Kim, S.Y.; Shin, J.; Oh, D.-C. Coprismycins A and B, neuroprotective phenylpyridines from the dung beetle-associated bacterium, Streptomyces sp. Bioorganic Med. Chem. Lett. 2011, 21, 5715–5718. [Google Scholar] [CrossRef]
- Park, S.-H.; Moon, K.; Bang, H.-S.; Kim, S.-H.; Kim, D.-G.; Oh, K.-B.; Shin, J.; Oh, D.-C. Tripartilactam, a Cyclobutane-Bearing Tricyclic Lactam from a Streptomyces sp. in a Dung Beetle’s Brood Ball. Org. Lett. 2012, 14, 1258–1261. [Google Scholar] [CrossRef] [PubMed]
- Kim, S.-H.; Kwon, S.H.; Park, S.-H.; Lee, J.K.; Bang, H.-S.; Nam, S.-J.; Kwon, H.C.; Shin, J.; Oh, D.-C. Tripartin, a Histone Demethylase Inhibitor from a Bacterium Associated with a Dung Beetle Larva. Org. Lett. 2013, 15, 1834–1837. [Google Scholar] [CrossRef]
- Um, S.; Bang, H.-S.; Shin, J.; Oh, D.-C. Actinofuranone C, a new 3-furanone-bearing polyketide from a dung beetle-associated bacterium. Nat. Prod. Sci. 2013, 19, 71–75. [Google Scholar]
- Um, S.; Shin, J.; Oh, K. New bioactive secondary metabolites from dung beetle-associated bacteria. Planta Medica 2016, 81, S1–S381. [Google Scholar] [CrossRef]
- Tyndale-Biscoe, M.; Wallace, M.M.H.; Walker, J.M. An ecological study of an Australian dung beetle, Onthophagus granulatus Boheman (Coleoptera: Scarabaeidae), using physiological age-grading techniques. Bull. Èntomol. Res. 1981, 71, 137–152. [Google Scholar] [CrossRef]
- Huerta, C.; Bang, H.S. Fecundity and Offspring Survival of Copris tripartitus Waterhouse (Coleoptera, Scarabaeidae: Scarabaeinae) Under Laboratory Rearing Conditions. Coleopt. Bull. 2004, 58, 501–507. [Google Scholar] [CrossRef]
- Lane, D.J. 16S/23S rRNA sequencing. In Nucleic Acid Techniques in Bacterial Systems; Jhon Wiley & Sons: Hoboken, NJ, USA, 1991. [Google Scholar]
- White, T.; Bruns, T.; Lee, S.; Taylor, J. Amplification and Direct Sequencing Of Fungal Ribosomal Rna Genes For Phylogenetics; Jhon Wiley & Sons: Hoboken, NJ, USA, 1990; pp. 315–322. [Google Scholar] [CrossRef]
- Hall, T.A. BioEdit: A user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acid Symp. Ser. 1999, 41, 95–98. [Google Scholar]
- Zhang, Z.; Schwartz, S.; Wagner, L.; Miller, W. A Greedy Algorithm for Aligning DNA Sequences. J. Comput. Biol. 2000, 7, 203–214. [Google Scholar] [CrossRef]
- Thompson, J.D.; Higgins, D.; Gibson, T.J. CLUSTAL W: Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 1994, 22, 4673–4680. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Kumar, S.; Tamura, K.; Nei, M. MEGA: Molecular Evolutionary Genetics Analysis software for microcomputers. Bioinformatics 1994, 10, 189–191. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Nei, M.; Kumar, S. Molecular Evolution and Phylogenetics; Oxford University Press: Oxford, UK, 2000. [Google Scholar]
- Thong, W.L.; Shin-Ya, K.; Nishiyama, M.; Kuzuyama, T. Methylbenzene-Containing Polyketides from a Streptomyces that Spontaneously Acquired Rifampicin Resistance: Structural Elucidation and Biosynthesis. J. Nat. Prod. 2016, 79, 857–864. [Google Scholar] [CrossRef]
- Barke, J.; Seipke, R.F.; Grüschow, S.; Heavens, D.; Drou, N.; Bibb, M.J.; Goss, R.J.; Yu, D.W.; I Hutchings, M. A mixed community of actinomycetes produce multiple antibiotics for the fungus farming ant Acromyrmex octospinosus. BMC Biol. 2010, 8, 109. [Google Scholar] [CrossRef] [Green Version]
- Heuer, H.; Krsek, M.; Baker, P.; Smalla, K.; Wellington, E.M. Analysis of actinomycete communities by specific amplification of genes encoding 16S rRNA and gel-electrophoretic separation in denaturing gradients. Appl. Environ. Microbiol. 1997, 63, 3233–3241. [Google Scholar] [CrossRef] [Green Version]
- Sheffield, V.C.; Cox, D.R.; Lerman, L.S.; Myers, R.M. Attachment of a 40-base-pair G + C-rich sequence (GC-clamp) to genomic DNA fragments by the polymerase chain reaction results in improved detection of single-base changes. Proc. Natl. Acad. Sci. USA 1989, 86, 232–236. [Google Scholar] [CrossRef] [Green Version]
- Sneath, P.H.A.; Sokal, R.R. Numerical Taxonomy. The Principles and Practice of Numerical Classification; W. H. Freeman and Co.: San Francisco, CA, USA, 1973. [Google Scholar]
- Labeda, D.P.; Dunlap, C.; Rong, X.; Huang, Y.; Doroghazi, J.R.; Ju, K.-S.; Metcalf, W.W. Phylogenetic relationships in the family Streptomycetaceae using multi-locus sequence analysis. Antonie van Leeuwenhoek 2016, 110, 563–583. [Google Scholar] [CrossRef] [PubMed]
- Mehling, A.; Wehmeier, U.F.; Piepersberg, W. Nucleotide sequences of streptomycete 16S ribosomal DNA: Towards a specific identification system for streptomycetes using PCR. Microbiology 1995, 141, 2139–2147. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Poulsen, M.; Oh, D.-C.; Clardy, J.; Currie, C.R. Chemical Analyses of Wasp-Associated Streptomyces Bacteria Reveal a Prolific Potential for Natural Products Discovery. PLoS ONE 2011, 6, e16763. [Google Scholar] [CrossRef] [Green Version]
- Oh, D.-C.; Poulsen, M.; Currie, C.R.; Clardy, J. Sceliphrolactam, a Polyene Macrocyclic Lactam from a Wasp-Associated Streptomyces sp. Org. Lett. 2011, 13, 752–755. [Google Scholar] [CrossRef] [PubMed]
- Leger, R.S.; Cooper, R.; Charnley, A. Cuticle-degrading enzymes of entomopathogenic fungi: Cuticle degradation in vitro by enzymes from entomopathogens. J. Invertebr. Pathol. 1986, 47, 167–177. [Google Scholar] [CrossRef]
- Butt, T.M.; Ibrahim, L.; Clark, S.J.; Beckett, A. The Germination Behavior of Metarhizium-Anisopliae on the Surface of Aphid and Flea Beetle Cuticles. Mycol. Res. 1995, 99, 945–950. [Google Scholar] [CrossRef]
- Steinkraus, D.C.; Geden, C.J.; Rutz, D.A. Susceptibility of Lesser Mealworm (Coleoptera, Tenebrionidae) to Beauveria-Bassiana (Moniliales, Moniliaceae)—Effects of Host Stage, Substrate, Formulation, and Host Passage. J. Med. Entomol. 1991, 28, 314–321. [Google Scholar] [CrossRef]
- Gindin, G.; Levski, S.; Glazer, I.; Soroker, V. Evaluation of the entomopathogenic fungiMetarhizium anisopliae andBeauveria bassiana against the red palm weevilRhynchophorus ferrugineus. Phytoparasitica 2006, 34, 370–379. [Google Scholar] [CrossRef]
- Hwang, J.-S.; Lee, J.; Kim, Y.-J.; Bang, H.-S.; Yun, E.-Y.; Kim, S.-R.; Suh, H.-J.; Kang, B.R.; Nam, S.-H.; Jeon, J.-P.; et al. Isolation and Characterization of a Defensin-Like Peptide (Coprisin) from the Dung Beetle, Copris tripartitus. Int. J. Pept. 2009, 2009, 1–5. [Google Scholar] [CrossRef]
- Lee, J.; Hwang, J.-S.; Hwang, I.-S.; Cho, J.; Lee, E.; Kim, Y.; Lee, D.G. Coprisin-induced antifungal effects in Candida albicans correlate with apoptotic mechanisms. Free Radic. Biol. Med. 2012, 52, 2302–2311. [Google Scholar] [CrossRef]
- Wang, Y.; Rozen, D.E. Gut Microbiota Colonization and Transmission in the Burying Beetle Nicrophorus vespilloides throughout Development. Appl. Environ. Microbiol. 2017, 83, e03250-16. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Duarte, A.; Welch, M.; Swannack, C.; Wagner, J.; Kilner, R.M. Strategies for managing rival bacterial communities: Lessons from burying beetles. J. Anim. Ecol. 2017, 87, 414–427. [Google Scholar] [CrossRef] [Green Version]
- Shukla, S.P.; Sanders, J.G.; Byrne, M.; Pierce, N.E. Gut microbiota of dung beetles correspond to dietary specializations of adults and larvae. Mol. Ecol. 2016, 25, 6092–6106. [Google Scholar] [CrossRef] [PubMed]
- Hwang, S.; Kim, E.; Lee, J.; Shin, J.; Yoon, Y.J.; Oh, D.-C. Structure Revision and the Biosynthetic Pathway of Tripartilactam. J. Nat. Prod. 2020, 83, 578–583. [Google Scholar] [CrossRef]
- Egonyu, J.P.; Torto, B. Responses of the ambrosia beetle Xylosandrus compactus (Coleoptera: Curculionidea: Scolytinae) to volatile constituents of its symbiotic fungus Fusarium solani (Hypocreales: Nectriaceae). Arthropod-Plant Interact. 2017, 12, 9–20. [Google Scholar] [CrossRef]
- Currie, C.R. A Community of Ants, Fungi, and Bacteria: A Multilateral Approach to Studying Symbiosis. Annu. Rev. Microbiol. 2001, 55, 357–380. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Kasson, M.T.; Wickert, K.L.; Stauder, C.M.; Macias, A.M.; Berger, M.C.; Simmons, D.R.; Short, D.P.; DeVallance, D.B.; Hulcr, J. Mutualism with aggressive wood-degrading Flavodon ambrosius (Polyporales) facilitates niche expansion and communal social structure in Ambrosiophilus ambrosia beetles. Fungal Ecol. 2016, 23, 86–96. [Google Scholar] [CrossRef] [Green Version]
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Kim, S.H.; Park, G.; Park, J.-S.; Kwon, H.C. Antifungal Streptomyces spp., Plausible Partners for Brood-Caring of the Dung Beetle Copris tripartitus. Microorganisms 2021, 9, 1980. https://doi.org/10.3390/microorganisms9091980
Kim SH, Park G, Park J-S, Kwon HC. Antifungal Streptomyces spp., Plausible Partners for Brood-Caring of the Dung Beetle Copris tripartitus. Microorganisms. 2021; 9(9):1980. https://doi.org/10.3390/microorganisms9091980
Chicago/Turabian StyleKim, Sung Hun, Goeun Park, Jin-Soo Park, and Hak Cheol Kwon. 2021. "Antifungal Streptomyces spp., Plausible Partners for Brood-Caring of the Dung Beetle Copris tripartitus" Microorganisms 9, no. 9: 1980. https://doi.org/10.3390/microorganisms9091980