Microbial Ecology

, Volume 54, Issue 2, pp 324–331 | Cite as

Culturable Bacteria Present in the Fluid of the Hooded-Pitcher Plant Sarracenia Minor Based on 16S rDNA Gene Sequence Data

  • Alex J. Siragusa
  • Janice E. Swenson
  • Dale A. Casamatta
Article

Abstract

The culturable microbial community within the pitcher fluid of 93 Sarracenia minor carnivorous plants was examined over a 2-year study. Many aspects of the plant/bacterial/insect interaction within the pitcher fluid are minimally understood because the bacterial taxa present in these pitchers have not been identified. Thirteen isolates were characterized by 16S rDNA sequencing and subsequent phylogenetic analysis. The Proteobacteria were the most abundant taxa and included representatives from Serratia, Achromobacter, and Pantoea. The Actinobacteria Micrococcus was also abundant while Bacillus, Lactococcus, Chryseobacterium, and Rhodococcus were infrequently encountered. Several isolates conformed to species identifiers (>98% rDNA gene sequence similarity) including Serratia marcescens (isolates found in 27.5% of pitchers), Achromobacterxylosoxidans (37.6%), Micrococcus luteus (40.9%), Bacillus cereus (isolates found in 10.2%), Bacillus thuringiensis (5.4%), Lactococcus lactis (17.2%), and Rhodococcusequi (2.2%). Species–area curves suggest that sampling efforts were sufficient to recover a representative culturable bacterial community. The bacteria present represent a diverse community probably as a result of introduction by insect vectors, but the ecological significance remains under explored.

References

  1. 1.
    Altayar, M, Sutherland, AD (2005) Bacillus cereus is common in the environment but emetic toxin producing isolates are rare. J Appl Microbiol 100: 7–14CrossRefGoogle Scholar
  2. 2.
    Aronson, NN, Halloran, BA, Alexyev, MF, Amable, L, Madura, JD, Pasupulati, L, Worth, C, Van Roey, P (2003) Family 18 chitinase-oligosaccharide substrate interaction: subsite preference and anomer selectivity of Serratia marcescens chitinase. A Biochem J 376: 87–95CrossRefGoogle Scholar
  3. 3.
    Arora, N, Ahmad, T, Rajagopal, R, Bhatnagar, RK (2003) A constitutively expressed 36kDa exochitinase from Bacillus thuringiensis HD-1. Biochem Biophys Res Commun 307: 620–625PubMedCrossRefGoogle Scholar
  4. 4.
    Belimov, AA, Hontzeas, N, Safronova, VI, Demchinskaya, SV, Piluzza, G, Bullitta, S, Glick, BR (2005) Cadmium-tolerant plant growth-promoting bacteria associated with the roots of Indian mustard (Brassica juncea L. Czern.). Soil Biol Biochem 37: 241–250CrossRefGoogle Scholar
  5. 5.
    Bernardet, JF, Vancanneyt, M, Matte-Tailliez, O, Grisez, L, Tailliez, P, Bizet, C, Nowakowski, M, Kerouault, B, Swings, J (2005) Polyphasic study of Chryseobacterium strains isolated from diseased aquatic animals. Syst Appl Microbiol 28: 640–660PubMedCrossRefGoogle Scholar
  6. 6.
    Bextine, B, Lampe, D, Lauzon, C, Jackson, B, Miller, TA (2005) Establishment of a genetically marked insect-derived symbiont in multiple host plants. Curr Microbiol 50: 1–7PubMedCrossRefGoogle Scholar
  7. 7.
    Boursaux-Eude, C, Gross, R (2000) New insights into symbiotic associations between ants and bacteria. Res Microbiol 151: 513–519PubMedCrossRefGoogle Scholar
  8. 8.
    Bradshaw, WE, Creelman, RA (1984) Mutualism between the carnivorous purple pitcher plant and its inhabitants. Am Midl Nat 112: 294–303CrossRefGoogle Scholar
  9. 9.
    Brandao, PFB, Clapp, JP, Bull, AT (2002) Discrimination and taxonomy of geographically diverse strains of nitrile-metabolizing actinomycetes using chemometric and molecular sequencing techniques. Environ Microbiol 4: 262–276PubMedCrossRefGoogle Scholar
  10. 10.
    Broderick, NA, Raffa, KF, Goodman, RM, Handelsman, J (2004) Census of the bacterial community of the Gypsy moth larval midgut by using culturing and culture-independent methods. Appl Environ Microbiol 70: 293–300CrossRefGoogle Scholar
  11. 11.
    Brosius, J, Palmer, ML, Kennedy, PJ, Noller, HF (1978) Complete nucleotide sequence of a 16S ribosomal RNA gene from Escherichia coli. Proc Natl Acad Sci USA 75: 4801–4805PubMedCrossRefGoogle Scholar
  12. 12.
    Cochran-Stafira, DL, von Ende, CN (1998) Integrating bacteria into food webs: studies with Sarracenia purpurea inquilines. Ecology 79: 880–898Google Scholar
  13. 13.
    Currie, CR, Bot, ANM, Boomsma, JJ (2003) Experimental evidence of a tripartite mutualism: bacteria protect ant fungus gardens from specialized parasites. Oikos 101: 92–102CrossRefGoogle Scholar
  14. 14.
    Dorman, HJD, Deans, SG (2000) Antimicrobial agents from plants: antibacterial activity of plant volatile oils. J Appl Microbiol 88: 308–316PubMedCrossRefGoogle Scholar
  15. 15.
    Dugas, JE, Zurek, L, Paster, BJ, Keddie, BA, Leadbetter, ER (2001) Isolation and characterization of a Chryseobacterium strain from the gut of the American cockroach, Periplaneta americana. Arch Microbiol 175: 259–262PubMedCrossRefGoogle Scholar
  16. 16.
    Ellison, AM, Farnsworth, EJ (2005) The cost of carnivory for Darlingtonia californica (Sarraceniaceae): Evidence from relationships among leaf traits. Am J Bot 92: 1085–1093Google Scholar
  17. 17.
    Feng, Y, Shen, D, Song, W (2006) Rice endophyte Pantoea agglomerans YS19 promotes host plant growth and affects allocations of host photosynthesis. J Appl Microbiol 100: 938–945PubMedCrossRefGoogle Scholar
  18. 18.
    Fish, D (1976) Insect-plant relationships of the insectivorous pitcher plant Sarracenia minor. Fla Entomol 2: 199–203Google Scholar
  19. 19.
    Germida, JJ, Siciliano, SD, Freitas, RD, Seib, AM (1998) Diversity of root-associated bacteria associated with field-grown canola (Brassica nuapus L.) and wheat (Triticum aestivum L.). FEMS Microbiol Ecol 26: 43–50CrossRefGoogle Scholar
  20. 20.
    Gitaitis, RD, Walcott, RR, Wells, ML, Diaz Perez, JC, Sanders, FH (2003) Transmission of Pantoea ananatis, causal agent of center rot of onion, by tobacco thrips, Frankliniella fusca. Plant Dis 87: 675–678CrossRefGoogle Scholar
  21. 21.
    Gohel, V, Megha, C, Vyas, P, Chhatpar, HS (2004) Strain improvement of chitinolytic enzyme producing isolate Pantoea dispersa for enhancing its biocontrol potential against fungal plant pathogens. Ann Microbiol 54: 503–515Google Scholar
  22. 22.
    Harvey, E, Miller, TE (1996) Variance in composition of inquiline communities in leaves of Sarracenia purpurea L. on multiple spatial scales. Oecologia 108: 562–566CrossRefGoogle Scholar
  23. 23.
    Hejazi, A, Falkiner, FR (1997) Serratia marcescens. J Med Microbiol 46: 903–912PubMedCrossRefGoogle Scholar
  24. 24.
    Hepburn, JS, St. John, EQ (1927) A bacteriological study of the pitcher liquor of the Sarraceniaceae. Trans Wagner Free Inst Sci Phila 11: 75–83Google Scholar
  25. 25.
    Horn, SJ, Sorbotten, A, Synstad, B, Sikorski, P, Sorlie, M, Varum, KM, Eijsink, VGH (2006) Endo/exo mechanism and processivity of family 18 chitinases produced by Serratia marcescens. FEBS J 273: 491–503PubMedCrossRefGoogle Scholar
  26. 26.
    Hui Xie, G, Zongjun, C, Yu, J, Yan, J, Hai, W, Steinberger, Y (2006) Identification of nif genes in N2-fixing bacterial strains isolated from rice fields along the Yangtze river plain. J Basic Microbiol 46: 56–63CrossRefGoogle Scholar
  27. 27.
    Lane, DJ (1991) 16S/23S rRNA sequencing. Nucleic acid techniques in bacterial systematics. Wiley, Chichester, United Kingdom, pp 115–175Google Scholar
  28. 28.
    Li, H, Medina, F, Vinson, SB, Coats, CJ (2005) Isolation, characterization, and molecular identification of bacteria from the red imported fire ant (Solenopsis invicta) midgut. J Invertebr Pathol 89: 203–209PubMedCrossRefGoogle Scholar
  29. 29.
    Liba, CM, Ferrara, FIS, Manfio, GP, Fantinatti-Garboggini, F, Albuquerque, RC, Pavan, C, Ramos, PL, Moreira-Filho, CA, Barbosa, HR (2006) Nitrogen-fixing chemo-organotrophic bacteria isolated from cyanobacteria-deprived lichens and their ability to release amino acids and phytohormones. J Appl Microbiol 101: 1076–1086PubMedCrossRefGoogle Scholar
  30. 30.
    Loranger-Merciris, G, Barthes, L, Gastine, A, Leadley, P (2006) Rapid effects of plant species diversity and identity on soil microbial communities in experimental grassland ecosystems. Soil Biol Biochem 38: 2336–2343CrossRefGoogle Scholar
  31. 31.
    Moran, NA, Telang, A (1998) Bacteriocyte-associated symbionts of insects. Bioscience 48: 295–304CrossRefGoogle Scholar
  32. 32.
    Muscatello, G, Browning, GF (2004) Identification and differentiation of avirulent and virulent Rhodococcus equi using selective media and colony blotting DNA hybridization to determine their concentrations in the environment. Vet Microbiol 100: 121–127PubMedCrossRefGoogle Scholar
  33. 33.
    Nomura, M, Kobayashi, M, Narita, T, Kimoto-Nira, H, Okamoto, T (2006) Phenotypic and molecular characterization of Lactococcus lactis from milk and plants. J Appl Microbiol 101: 396–405PubMedCrossRefGoogle Scholar
  34. 34.
    Pankhurst, CE, Ophel-Keller, K, Doube, BM, Gupta, VVSR (1996) Biodiversity of soil microbial communities in agricultural systems. Biodivers Conserv 5: 197–209CrossRefGoogle Scholar
  35. 35.
    Plummer, GL, Jackson, TH (1963) Bacterial activities within the sarcophagus of the insectivorous plant, Sarracenia flava. Am Midl Nat 69: 462–469CrossRefGoogle Scholar
  36. 36.
    Posada, D, Crandall, KA (1998) MODELTEST: testing the model of DNA substitution. Bioinformatics 14: 817–818PubMedCrossRefGoogle Scholar
  37. 37.
    Prankevicius, AB, Cameron, DM (1991) Bacterial dinitrogen fixation in the leaf of the northern pitcher plant (Sarracenia purpurea). Can J Bot 69: 2296–2298Google Scholar
  38. 38.
    Schnepf, E, Crickmore, N, Van Rie, J, Lereclus, D, Baum, J, Feitelson, J, Zeigler, DR, Dean, DH (1998) Bacillus thuringiensis and its pesticidal crystal proteins. Microbiol Mol Biol Rev 62: 775–806PubMedGoogle Scholar
  39. 39.
    Schreiber, L, Krimm, U, Knoll, D, Sayed, M, Auling, G, Kroppenstedt, RM (2005) Plant-microbe interactions: identification of epiphytic bacteria and their ability to alter leaf surface permeability. New Phytol 166: 589–594PubMedCrossRefGoogle Scholar
  40. 40.
    Swofford, DL (1998) PAUP—Phylogenetic Analysis Using Parsimony, ver. 4.02. Sinaur Associates, Sunderland, MassachusettsGoogle Scholar
  41. 41.
    Thompson, JD, Gibson, TJ, Plewniak, F, Jeanmougin, F, Higgins, DG (1997) The Clustal X-windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25: 4876–4882PubMedCrossRefGoogle Scholar
  42. 42.
    Wang, G, Skipper, HD (2004) Identification of denitrifying rhizobacteria from bentgrass and bermuda grass golf greens. J Appl Microbiol 97: 827–837PubMedCrossRefGoogle Scholar
  43. 43.
    Wellington, EMH, Berry, A, Krsek, M (2003) Resolving functional diversity in relation to microbial community structure in soil: exploiting genomics and stable isotope probing. Curr Opin Microbiol 6: 295–301PubMedCrossRefGoogle Scholar
  44. 44.
    Whitman, RL, Byers, SE, Shively, DA, Ferguson, DM, Byappanahalli, M (2005) Occurrence and growth characterisitics of Escherichia coli and enterococci within the accumulated fluid of the northern pitcher plant (Sarracenia purpurea L.). Can J Microbiol 51: 1027–1037PubMedCrossRefGoogle Scholar
  45. 45.
    Wolfe, LM (1981) Feeding behavior of a plant: Differential prey capture in old and new leaves of the pitcher plant (Sarracenia purpurea). Am Midl Nat 106: 352–359CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2007

Authors and Affiliations

  • Alex J. Siragusa
    • 1
  • Janice E. Swenson
    • 1
  • Dale A. Casamatta
    • 1
  1. 1.Department of BiologyUniversity of North FloridaJacksonvilleUSA

Personalised recommendations