Isolation and Identification of Cellulolytic Bacteria from the Gut of Holotrichia parallela Larvae (Coleoptera: Scarabaeidae)

Abstract

In this study, 207 strains of aerobic and facultatively anaerobic cellulolytic bacteria were isolated from the gut of Holotrichia parallela larvae. These bacterial isolates were assigned to 21 genotypes by amplified ribosomal DNA restriction analysis (ARDRA). A partial 16S rDNA sequence analysis and standard biochemical and physiological tests were used for the assignment of the 21 representative isolates. Our results show that the cellulolytic bacterial community is dominated by the Proteobacteria (70.05%), followed by the Actinobacteria (24.15%), the Firmicutes (4.35%), and the Bacteroidetes (1.45%). At the genus level, Gram-negative bacteria including Pseudomonas, Ochrobactrum, Rhizobium, Cellulosimicrobium, and Microbacterium were the predominant groups, but members of Bacillus, Dyadobacter, Siphonobacter, Paracoccus, Kaistia, Devosia, Labrys, Ensifer, Variovorax, Shinella, Citrobacter, and Stenotrophomonas were also found. Furthermore, our results suggest that a significant amount of bacterial diversity exists among the cellulolytic bacteria, and that Siphonobacter aquaeclarae, Cellulosimicrobium funkei, Paracoccus sulfuroxidans, Ochrobactrum cytisi, Ochrobactrum haematophilum, Kaistia adipata, Devosia riboflavina, Labrys neptuniae, Ensifer adhaerens, Shinella zoogloeoides, Citrobacter freundii, and Pseudomonas nitroreducens are reported to be cellulolytic for the first time in this study. Our results indicate that the scarab gut is an attractive source for the study of novel cellulolytic microorganisms and enzymes useful for cellulose degradation.

References 69

1. 1.

   Sun J.Z., Scharf M.E. Exploring and integrating cellulolytic systems of insects to advance biofuel technology. Insect Sci. 2010;17:163-165.

   Not in our library

2. 2.

   Lynd L.R., Cushman J.H., Nichols R.J., Wyman C.E. Fuel ethanol from cellulosic biomass. Science. 1991;251:1318-1323.

   Not in our library10.1126/science.251.4999.1318

3. 3.

   Lynd L.R., Laser M.S., Bransby D., Dale B.E., Davison B., Hamilton R., Himmel M., Keller M., McMillan J.D., Sheehan J., et al. How biotech can transform biofuels. Nat. Biotechnol. 2008;26:169-172.

   Not in our library10.1038/nbt0208-169

4. 4.

   Kim N., Choo Y.M., Lee K.S., Hong S.J., Seol K.Y., Je Y.H., Sohn H.D., Jin B.R. Molecular cloning and characterization of a glycosyl hydrolase family 9 cellulase distributed throughout the digestive tract of the cricket Teleogryllus emma. Comp. Biochem. Physiol. B Biochem. Mol. Biol. 2008;150:368-376.

   Not in our library10.1016/j.cbpb.2008.04.005

5. 5.

   Sánchez Ó.J., Cardona C.A. Trends in biotechnological production of fuel ethanol from different feedstocks. Bioresour. Technol. 2008;99:5270-5295.

   Not in our library10.1016/j.biortech.2007.11.013

6. 6.

   Sanderson K. Us biofuels: A field in ferment. Nature. 2006;444:673-676.

   Not in our library10.1038/444673a

7. 7.

   Badger P.C., Janick J., Whipkey A. Ethanol from Cellulose: A General Review. Trends in New Crops and New Uses. 2002;17-21.

   Not in our library

8. 8.

   Hamelinck C.N., van Hooijdonk G., Faaij A.P.C. Ethanol from lignocellulosic biomass: Techno-economic performance in short-, middle- and long-term. Biomass Bioenergy. 2005;28:384-410.

   Not in our library

9. 9.

   Martin M., Ahmetovic E., Grossmann I.E. Optimization of water consumption in second generation bioethanol plants. Ind. Eng. Chem. Res. 2010;50:3705-3721.

   Not in our library

10. 10.

    Mabee W.E., Saddler J.N. Bioethanol from lignocellulosics: Status and perspectives in Canada. Bioresour. Technol. 2010;101:4806-4813.

    Not in our library10.1016/j.biortech.2009.10.098

11. 11.

    Demirbas A. Options and trends of thorium fuel utilization in turkey. Energy Sources. 2005;27:597-603.

    Not in our library

12. 12.

    Balat M., Balat H., Öz C. Progress in bioethanol processing. Prog. Energy Combust. Sci. 2008;34:551-573.

    Not in our library

13. 13.

    Eriksson T., Börjesson J., Tjerneld F. Mechanism of surfactant effect in enzymatic hydrolysis of lignocellulose. Enzyme Microb. Technol. 2002;31:353-364.

    Not in our library

14. 14.

    Ferreira S., Duarte A.P., Ribeiro M.H.L., Queiroz J.A., Domingues F.C. Response surface optimization of enzymatic hydrolysis of cistus ladanifer and cytisus striatus for bioethanol production. Biochem. Eng. J. 2009;45:192-200.

    Not in our library

15. 15.

    Eijsink V.G.H., Vaaje-Kolstad G., Vårum K.M., Horn S.J. Towards new enzymes for biofuels: Lessons from chitinase research. Trends Biotechnol. 2008;26:228-235.

    Not in our library10.1016/j.tibtech.2008.02.004

16. 16.

    Mojović L., Nikolić S., Rakin M., Vukasinović M. Production of bioethanol from corn meal hydrolyzates. Fuel. 2006;85:1750-1755.

    Not in our library

17. 17.

    Piskur J., Rozpedowska E., Polakova S., Merico A., Compagno C. How did Saccharomyces evolve to become a good brewer? Trends Genet. 2006;22:183-186.

    Not in our library10.1016/j.tig.2006.02.002

18. 18.

    Alper1 H., Stephanopoulos G. Engineering for biofuels: Exploiting innate microbial capacity or importing biosynthetic potential? Nat. Rev. Microbiol. 2009;7:715-723.

    Not in our library10.1038/nrmicro2186

19. 19.

    Zhou X., Smith J.A., Oi F.M., Koehler P.G., Bennett G.W., Scharf M.E. Correlation of cellulase gene expression and cellulolytic activity throughout the gut of the termite Reticulitermes flavipes. Gene. 2007;395:29-39.

    Not in our library10.1016/j.gene.2007.01.004

20. 20.

    Geib S.M., Tien M., Hoover K. Identification of proteins involved in lignocellulose degradation using in gel zymogram analysis combined with mass spectroscopy-based peptide analysis of gut proteins from larval asian longhorned beetles, Anoplophora glabripennis. Insect Sci. 2010;17:253-264.

    Not in our library

21. 21.

    Ferreira A.H., Marana S.R., Terra W.R., Ferreira C. Purification, molecular cloning, and properties of a beta-glycosidase isolated from midgut lumen of Tenebrio molitor (Coleoptera) larvae. Insect Biochem. Mol. Biol. 2001;31:1065-1076.

    Not in our library10.1016/s0965-1748(01)00054-6

22. 22.

    Cazemier A.E., Op den Camp H.J.M., Hackstein J.H.P., Vogels G.D. Fibre digestion in arthropods. Comp. Biochem. Physiol. A Physiol. 1997;118:101-109.

    Not in our library

23. 23.

    Brune A. Termite guts: The world’s smallest bioreactors. Trends Biotechnol. 1998;16:16-21.

    Not in our library

24. 24.

    Breznak J.A., Brune A. Role of microorganisms in the digestion of lignocellulose by termites. Annu. Rev. Entomol. 1994;39:453-487.

    Not in our library

25. 25.

    Wenzel M., Schonig I., Berchtold M., Kampfer P., Konig H. Aerobic and facultatively anaerobic cellulolytic bacteria from the gut of the termite Zootermopsis angusticollis. J. Appl. Microbiol. 2002;92:32-40.

    Not in our library10.1046/j.1365-2672.2002.01502.x

26. 26.

    Watanabe H., Tokuda G. Cellulolytic systems in insects. Annu. Rev. Entomol. 2010;55:609-632.

    Not in our library10.1146/annurev-ento-112408-085319

27. 27.

    Chakraborty N., Sarkar G.M., Lahiri S.C. Cellulose degrading capabilities of cellulolytic bacteria isolated from the intestinal fluids of the silver cricket. Environmentalist. 2000;20:9-11.

    Not in our library

28. 28.

    Cook D.M., Doran-Peterson J. Mining diversity of the natural biorefinery housed within Tipula abdominalis larvae for use in an industrial biorefinery for production of lignocellulosic ethanol. Insect Sci. 2010;17:303-312.

    Not in our library

29. 29.

    Delalibera I., Handelsman J., Raffa K.F. Contrasts in cellulolytic activities of gut microorganisms between the wood borer, Saperda vestita (Coleoptera: Cerambycidae), and the bark beetles, Ips pini and Dendroctonus frontalis (Coleoptera: Curculionidae). Environ. Entomol. 2005;34:541-547.

    Not in our library

30. 30.

    Cazemier A.E., Verdoes J.C., Reubsaet F.A., Hackstein J.H., van der Drift C., Op den Camp H.J. Promicromonospora pachnodae sp. nov., a member of the (hemi)cellulolytic hindgut flora of larvae of the scarab beetle Pachnoda marginata. Antonie Van Leeuwenhoek. 2003;83:135-148.

    Not in our library10.1023/a:1023325817663

31. 31.

    Huang S.W., Zhang H.Y., Marshall S., Jackson T.A. The scarab gut: A potential bioreactor for bio-fuel production. Insect Sci. 2010;17:175-183.

    Not in our library

32. 32.

    Lavelle P., Bignell D., Lepage M., Wolters V., Roger P., Ineson P., Heal O.W., Dhillion S. Soil function in a changing world: The role of invertebrate ecosystem engineers. Eur. J. Soil Biol. 1997;33:159-193.

    Not in our library

33. 33.

    Cazemier A.E., Hackstein J.H.P., Op den Camp H.J.M., Rosenberg J., van der Drift C. Bacteria in the intestinal tract of different species of arthropods. Microb. Ecol. 1997;33:189-197.

    Not in our library10.1007/s002489900021

34. 34.

    Zhang H.Y., Jackson T.A. Autochthonous bacterial flora indicated by PCR-DGGE of 16S rRNA gene fragments from the alimentary tract of Costelytra zealandica (Coleoptera: Scarabaeidae). J. Appl. Microbiol. 2008;105:1277-1285.

    Not in our library10.1111/j.1365-2672.2008.03867.x

35. 35.

    Bayon C., Mathelin J. Carbohydrate fermentation and by-product absorption studied with labeled cellulose in Oryctes nasicornis larvae (Coleoptera: Scarabaeidae). J. Insect Physiol. 1980;26:833-840.

    Not in our library

36. 36.

    Geissinger O., Herlemann D.P.R., Mörschel E., Maier U.G., Brune A. The ultramicrobacterium “Elusimicrobium minutum” gen. nov., sp. nov., the first cultivated representative of the termite group 1 phylum. Appl. Environ. Microbiol. 2009;75:2831-2840.

    Not in our library10.1128/aem.02697-08

37. 37.

    Zhou L.M., Ju Q., Qu M.J., Zhao Z.Q., Dong S.L., Han Z.J., Yu S.L. EAG and behavioral responses of the large black chafer, Holotrichia parallela (Coleoptera: Scarabaeidae) to its sex pheromone. Acta Entomol. Sin. 2009;52:121-125.

    Not in our library

38. 38.

    Egert M., Wagner B., Lemke T., Brune A., Friedrich M.W. Microbial community structure in midgut and hindgut of the humus-feeding larva of Pachnoda ephippiata (Coleoptera: Scarabaeidae). Appl. Environ. Microbiol. 2003;69:6659-6668.

    Not in our library10.1128/aem.69.11.6659-6668.2003

39. 39.

    Egert M., Stingl U., Bruun D.L., Wagner B., Brune A., Friedrich M.W. Structure and topology of microbial communities in the major gut compartments of Melolontha melolontha larvae (Coleoptera: Scarabaeidae). Appl. Environ. Microbiol. 2005;71:4556-4566.

    Not in our library10.1128/aem.71.8.4556-4566.2005

40. 40.

    Lemke T., Stingl U., Egert M., Friedrich M.W., Brune A. Physicochemical conditions and microbial activities in the highly alkaline gut of the humus-feeding larva of Pachnoda ephippiata (Coleoptera: Scarabaeidae). Appl. Environ. Microbiol. 2003;69:6650-6658.

    Not in our library10.1128/aem.69.11.6650-6658.2003

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