Browse > Article
http://dx.doi.org/10.7845/kjm.2015.5018

Analysis of gut bacterial diversity and exploration of cellulose-degrading bacteria in xylophagous insects  

Choi, Min-Young (Agricultural Microbiology Division, National Academy of Agricultural Science (NAAS), Rural Development Administration (RDA))
Ahn, Jae-Hyung (Agricultural Microbiology Division, National Academy of Agricultural Science (NAAS), Rural Development Administration (RDA))
Song, Jaekyeong (Agricultural Microbiology Division, National Academy of Agricultural Science (NAAS), Rural Development Administration (RDA))
Kim, Seong-Hyun (Applied Entomology Division, NAAS, RDA)
Bae, Jin-Woo (Department of Biology, Kyung Hee University)
Weon, Hang-Yeon (Agricultural Microbiology Division, National Academy of Agricultural Science (NAAS), Rural Development Administration (RDA))
Publication Information
Korean Journal of Microbiology / v.51, no.3, 2015 , pp. 209-220 More about this Journal
Abstract
In this study, gut bacterial communities in xylophagous insects were analyzed using the pyrosequencing of 16S rRNA genes for their potential biotechnological applications in lignocelluloses degradation. The result showed that operational taxonomic units (OTUs), species richness and diversity index were higher in the hindgut than in the midgut of all insect samples analyzed. The dominant phyla or classes were Firmicutes (54.0%), Bacteroidetes (14.5%), ${\gamma}-Proteobacteria$ (12.3%) in all xylophagous insects except for Rhinotermitidae. The principal coordinates analysis (PCoA) showed that the bacterial community structure mostly clustered according to phylogeny of hosts rather than their habitats. In our study, the two carboxymethyl cellulose (CMC)-degrading isolates which showed the highest enzyme activity were most closely related to Bacillus toyonensis $BCT-7112^T$ and Lactococcus lactis subsp. hordniae $NCDO\;2181^T$, respectively. Cellulolytic enzyme activity analysis showed that ${\beta}-1,4-glucosidase$, ${\beta}-1,4-endoglucanase$ and ${\beta}-1,4-xylanase$ were higher in the hindgut of Cerambycidae. The results demonstrate that xylophagous insect guts harbor diverse gut bacteria, including valuable cellulolytic bacteria, which could be used for various biotechnological applications.
Keywords
cellulolytic bacteria; lignocellulose degradation; pyrosequencing; xylophagous insect;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Arias-Cordero, E., Ping, L.Y., Reichwald, K., Delb, H., Platzer, M., and Boland, W. 2012. Comparative evaluation of the gut microbiota associated with the below- and above-ground life stages (larvae and beetles) of the forest cockchafer, Melolontha hippocastani. PLoS ONE 7, e51557.   DOI
2 Bradford, M.M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248-254.   DOI   ScienceOn
3 Brune, A. 2014. Symbiotic digestion of lignocellulose in termite guts. Nat. Rev. Microbiol. 12, 168-180.   DOI
4 Cazemier, A.E., Verdoes, J.C., Reubsaet, F.A.G., Hackstein, J.H.P., van der Drift, C., and den Camp, H. 2003. Promicromonospora pachnodae sp. nov., a member of the (hemi)cellulolytic hindgut flora of larvae of the scarab beetle Pachnoda marginata. Antonie van Leeuwenhoek 83, 135-148.   DOI
5 Chun, J., Kim, K.Y., Lee, J.H., and Choi, Y. 2010. The analysis of oral microbial communities of wild-type and toll-like receptor 2-deficient mice using a 454 GS FLX Titanium pyrosequencer. BMC Microbiol. 10, 101.   DOI   ScienceOn
6 Clemente, J.C., Ursell, L.K., Parfrey, L.W., and Knight, R. 2012. The impact of the gut microbiota on human health: an integrative view. Cell. 148, 1258-1270.   DOI   ScienceOn
7 Cole, J.R., Wang, Q., Cardenas, E., Fish, J., Chai, B., Farris, R.J., Kulam-Syed-Mohideen, A.S., McGarrell, D.M., Marsh, T., Garrity, G.M., et al. 2009. The Ribosomal Database Project:improved alignments and new tools for rRNA analysis. Nucleic Acids Res. 37, D141-D145.   DOI
8 Colman, D.R., Toolson, E.C., and Takacs-Vesbach, C.D. 2012. Do diet and taxonomy influence insect gut bacterial communities? Mol. Ecol. 21, 5124-5137.   DOI
9 Cook, D.M. and Doran-Peterson, J. 2010. Mining diversity of the natural biorefinery housed within Tipula abdominalis larvae for use in an industrial biorefinery for production of lignocellulosic ethanol. Insect Sci. 17, 303-312.   DOI
10 Despres, L., David, J.P., and Gallet, C. 2007. The evolutionary ecology of insect resistance to plant chemicals. Trends Ecol. Evol. 22, 298-307.   DOI
11 Dillon, R. and Charnley, K. 2002. Mutualism between the desert locust Schistocerca gregaria and its gut microbiota. Res. Microbiol. 153, 503-509.   DOI
12 Dillon, R.J. and Dillon, V.M. 2004. The gut bacteria of insects:Nonpathogenic interactions. Annu. Rev. Entomol. 49, 71-92.   DOI
13 Edgar, R.C. 2013. UPARSE: highly accurate OTU sequences from microbial amplicon reads. Nat. Methods 10, 996-998.   DOI
14 Eijsink, V.G.H., Vaaje-Kolstad, G., Varum, K.M., and Horn, S.J. 2008. Towards new enzymes for biofuels: lessons from chitinase research. Trends Biotechnol. 26, 228-235.   DOI
15 Engel, P. and Moran, N.A. 2013. The gut microbiota of insects - diversity in structure and function. FEMS Microbiol. Rev. 37, 699-735.   DOI
16 Eriksson, T., Borjesson, J., and Tjerneld, F. 2002. Mechanism of surfactant effect in enzymatic hydrolysis of lignocellulose. Enzyme Microb. Technol. 31, 353-364.   DOI
17 Farrell, A.E., Plevin, R.J., Turner, B.T., Jones, A.D., O'Hare, M., and Kammen, D.M. 2006. Ethanol can contribute to energy and environmental goals. Science 311, 506-508.   DOI   ScienceOn
18 Geib, S.M., Jimenez-Gasco, M.D.M., Carlson, J.E., Tie, M., and Hoover, K. 2009. Effect of host tree species on cellulase activity and bacterial community composition in the gut of larval asian longhorned beetle. Environ. Entomol. 38, 686-699.   DOI
19 Gruenwald, S., Pilhofer, M., and Hoell, W. 2010. Microbial associations in gut systems of wood- and bark-inhabiting longhorned beetles [Coleoptera: Cerambycidae]. Syst. Appl. Microbiol. 33, 25-34.   DOI
20 Grieco, M.A., Cavalcante, J.J., Cardoso, A.M., Vieira, R.P., Machado, E.A., Clementino, M.M., Medeiros, M.N., Albano, R.M., Garcia, E.S., de Souza, W., et al. 2013. Microbial community diversity in the gut of the south American termite Cornitermes cumulans (Isoptera: Termitidae). Microb. Ecol. 65, 197-204.   DOI
21 Hahn-Hagerdal, B., Galbe, M., Gorwa-Grauslund, M.F., Liden, G., and Zacchi, G. 2006. Bio-ethanol - the fuel of tomorrow from the residues of today. Trends Biotechnol. 24, 549-556.   DOI
22 Hamelinck, C.N., van Hooijdonk, G., and Faaij, A.P.C. 2005. Ethanol from lignocellulosic biomass: techno-economic performance in short-, middle- and long-term. Biomass Bioenerg. 28, 384-410.   DOI
23 Hebert, P.D.N., Cywinska, A., Ball, S.L., and DeWaard, J.R. 2003. Biological identifications through DNA barcodes. Proc. R. Soc. B-Biol. Sci. 270, 313-321.   DOI
24 Huang, X.F., Bakker, M.G., Judd, T.M., Reardon, K.F., and Vivanco, J.M. 2013. Variations in diversity and richness of gut bacterial communities of termites (Reticulitermes flavipes) fed with grassy and woody plant substrates. Microb. Ecol. 65, 531-536.   DOI
25 Huang, S.W., Zhang, H.Y., Marshall, S., and Jackson, T.A. 2010. The scarab gut: A potential bioreactor for bio-fuel production. Insect Sci. 17, 175-183.   DOI
26 Kaufman, M.G. and Klug, M.J. 1991. The contribution of hindgut bacteria to dietary carbohydrate utilization by crickets (Orthoptera, Gryllidae). Comp. Biochem. Physiol. 98, 117-123.   DOI
27 Kukor, J.J., Cowan, D.P., and Martin, M.M. 1988. The role of ingested fungal enzymes in cellulose digestion in the larvae of cerambycid beetles. Physiol. Zool. 61, 364-371.   DOI
28 Kim, O.S., Cho, Y.J., Lee, K., Yoon, S.H., Kim, M., Na, H., Park, S.C., Jeon, Y.S., Lee, J.H., Yi, H., et al. 2012. Introducing EzTaxon-e:A prokaryotic 16S rRNA gene sequence database with phylotypes that represent uncultured species. Int. J. Syst. Evol. Microbiol. 62, 716-721.   DOI
29 Konig, H., Li, L., and Frohlich, J. 2013. The cellulolytic system of the termite gut. Appl. Microbiol. Biotechnol. 97, 7943-7962.   DOI
30 Koroiva, R., Souza, C.W.O., Toyama, D., Henrique-Silva, F., and Fonseca-Gessner, A.A. 2013. Lignocellulolytic enzymes and bacteria associated with the digestive tracts of Stenochironomus (Diptera: Chironomidae) larvae. Genet. Mol. Res. 12, 3421-3434.
31 Lykidis, A., Mavromatis, K., Ivanova, N., Anderson, I., Land, M., DiBartolo, G., Martinez, M., Lapidus, A., Lucas, S., Copeland, A., et al. 2007. Genome sequence and analysis of the soil cellulolytic actinomycete Thermobifida fusca YX. J. Bacteriol. 189, 2477-2486.   DOI
32 Maki, M., Leung, K.T., and Qin, W.S. 2009. The prospects of cellulaseproducing bacteria for the bioconversion of lignocellulosic biomass. Int. J. Biol. Sci. 5, 500-516.
33 Menon, V. and Rao, M. 2012. Trends in bioconversion of lignocellulose: Biofuels, platform chemicals & biorefinery concept. Prog. Energy Combust. Sci. 38, 522-550.   DOI   ScienceOn
34 Miller, G.L. 1959. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal. Chem. 31, 426-428.   DOI
35 Schauer, C., Thompson, C.L., and Brune, A. 2012. The bacterial community in the gut of the cockroach Shelfordella lateralis reflects the close evolutionary relatedness of cockroaches and termites. Appl. Environ. Microbiol. 78, 2758-2767.   DOI
36 Park, D.S., Oh, H.W., Jeong, W.J., Kim, H., Park, H.Y., and Bae, K.S. 2007. A culture-based study of the bacterial communities within the guts of nine longicorn beetle species and their exo-enzyme producing properties for degrading xylan and pectin. J. Microbiol. 45, 394-401.
37 Reid, N.M., Addison, S.L., Macdonald, L.J., and Lloyd-Jones, G. 2011. Biodiversity of active and inactive bacteria in the gut flora of wood-feeding huhu beetle larvae (Prionoplus reticularis). Appl. Environ. Microbiol. 77, 7000-7006.   DOI
38 Schauer, C., Thompson, C., and Brune, A. 2014. Pyrotag sequencing of the gut microbiota of the cockroach Shelfordella lateralis reveals a highly dynamic core but only limited effects of diet on community structure. PLoS ONE 9, e85861.   DOI
39 Schloss, P.D., Delalibera, I., Handelsman, J., and Raffa, K.F. 2006. Bacteria associated with the guts of two wood-boring beetles: Anoplophora glabripennis and Saperda vestita (Cerambycidae). Environ. Entomol. 35, 625-629.   DOI
40 Schloss, P.D., Westcott, S.L., Ryabin, T., Hall, J.R., Hartmann, M., Hollister, E.B., Lesniewski, R.A., Oakley, B.B., Parks, D.H., Robinson, C.J., et al. 2009. Introducing mothur: Open-source, platform-independent, community-supported software for describing and comparing microbial communities. Appl. Environ. Microbiol. 75, 7537-7541.   DOI
41 Shi, W., Ding, S.Y., and Yuan, J.S. 2011. Comparison of insect gut cellulase and xylanase activity across different insect species with distinct food sources. Bioenerg. Res. 4, 1-10.   DOI
42 Wang, A.L., Yao, Z.C., Zheng, W.W., and Zhang, H.Y. 2014. Bacterial communities in the gut and reproductive organs of Bactrocera minax (Diptera: Tephritidae) based on 454 pyrosequencing. PLoS ONE 9, e106988.   DOI
43 Sims, R.E.H., Mabee, W., Saddler, J.N., and Taylor, M. 2010. An overview of second generation biofuel technologies. Bioresour. Technol. 101, 1570-1580.   DOI
44 Sudakaran, S., Salem, H., Kost, C., and Kaltenpoth, M. 2012. Geographical and ecological stability of the symbiotic mid-gut microbiota in European firebugs, Pyrrhocoris apterus (Hemiptera, Pyrrhocoridae). Mol. Ecol. 21, 6134-6151.   DOI
45 Sun, J.Z. and Scharf, M.E. 2010. Exploring and integrating cellulolytic systems of insects to advance biofuel technology. Insect Sci. 17, 163-165.   DOI
46 Warnecke, F., Luginbuhl, P., Ivanova, N., Ghassemian, M., Richardson, T.H., Stege, J.T., Cayouette, M., McHardy, A.C., Djordjevic, G., Aboushadi, N., et al. 2007. Metagenomic and functional analysis of hindgut microbiota of a wood-feeding higher termite. Nature 450, 560-565.   DOI
47 Watanabe, H. and Tokuda, G. 2010. Cellulolytic systems in insects. Annu. Rev. Entomol. 55, 609-632.   DOI
48 Xu, J. and Gordon, J.I. 2003. Honor thy symbionts. Proc. Natl. Acad. Sci. USA 100, 10452-10459.   DOI   ScienceOn
49 Yun, J.H., Roh, S.W., Whon, T.W., Jung, M.J., Kim, M.S., Park, D.S., Yoon, C., Nam, Y.D., Kim, Y.J., Choi, J.H., et al. 2014. Insect gut bacterial diversity determined by environmental habitat, diet, developmental stage, and phylogeny of host. Appl. Environ. Microbiol. 80, 5254-5264.   DOI