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Microbial Ecology

, Volume 62, Issue 1, pp 188–197 | Cite as

Endophytes of Grapevine Flowers, Berries, and Seeds: Identification of Cultivable Bacteria, Comparison with Other Plant Parts, and Visualization of Niches of Colonization

  • Stéphane CompantEmail author
  • Birgit Mitter
  • Juan Gualberto Colli-Mull
  • Helmut Gangl
  • Angela Sessitsch
Plant Microbe Interactions

Abstract

Endophytic bacteria can colonize various plants and organs. However, endophytes colonizing plant reproductive organs have been rarely analyzed. In this study, endophytes colonizing flowers as well as berries and seeds of grapevine plants grown under natural conditions were investigated by cultivation as well as by fluorescence in situ hybridization. For comparison, bacteria were additionally isolated from other plant parts and the rhizosphere and characterized. Flowers, fruits, and seeds hosted various endophytic bacteria. Some taxa were specifically isolated from plant reproductive organs, whereas others were also detected in the rhizosphere, endorhiza or grape inflo/infructescence stalk at the flowering or berry harvest stage. Microscopic analysis by fluorescence in situ hybridization of resin-embedded samples confirmed the presence of the isolated taxa in plant reproductive organs and enabled us to localize them within the plant. Gammaproteobacteria (including Pseudomonas spp.) and Firmicutes (including Bacillus spp.) were visualized inside the epidermis and xylem of ovary and/or inside flower ovules. Firmicutes, mainly Bacillus spp. were additionally visualized inside berries, in the intercellular spaces of pulp cells and/or xylem of pulp, but also along some cell walls inside parts of seeds. Analysis of cultivable bacteria as well as microscopic results indicated that certain endophytic bacteria can colonize flowers, berries, or seeds. Our results also indicated that some specific taxa may not only derive from the root environment but also from other sources such as the anthosphere.

Keywords

Plant Part Firmicutes Gammaproteobacteria Endophytic Bacterium Xylem Vessel 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

We are grateful to Anton Grashl (Seibersdorf, Austria) for the help with sampling. We additionally thank Dr. Elsa Arcalis from the University of Natural Resources and Life Sciences of Vienna (Austria) for allowing use of a microtome as well as glass knives for plant sections. This work was supported by a Hertha Firnberg program from AIT.

References

  1. 1.
    Altschul SF, Madden TL, Schäffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402PubMedCrossRefGoogle Scholar
  2. 2.
    Amann RI, Binder BJ, Olson RJ, Chisholm SW, Devereux R, Stahl DA (1990) Combination of 16S rRNA-targeted oligonucleotide probes with flow cytometry for analyzing mixed microbial populations. Appl Environ Microbiol 56:1919–1925PubMedGoogle Scholar
  3. 3.
    Amann R, Ludwig W, Schulze R, Spring S, Moore E, Schleifer K-H (1996) rRNA-targeted oligonucleotide probes for the identification of genuine and former pseudomonads. Syst Appl Microbiol 19:501–509Google Scholar
  4. 4.
    Barac T, Taghavi S, Borremans B, Provoost A, Oeyen L, Colpaert JV, Vangronsveld J, van der Lelie D (2004) Engineered endophytic bacteria improve phytoremediation of water-soluble, volatile, organic pollutants. Nat Biotechnol 22:583–588PubMedCrossRefGoogle Scholar
  5. 5.
    Bell CR, Dickie GA, Harvey WLG, Chan JWYF (1995) Endophytic bacteria in grapevine. Can J Microbiol 41:46–53CrossRefGoogle Scholar
  6. 6.
    Bulgari D, Casati P, Brusetti L, Quaglino F, Brasca M, Daffonchio D, Bianco PA (2009) Endophytic bacterial diversity in grapevine (Vitis vinifera L.) leaves described by 16S rRNA gene sequence analysis and length heterogeneity-PCR. J Microbiol 47:393–401PubMedCrossRefGoogle Scholar
  7. 7.
    Cankar K, Kraigher H, Ravnikar M, Rupnik M (2005) Bacterial endophytes from seeds of Norway spruce (Picea abies L. Karst). FEMS Microbiol Lett 244:341–345PubMedCrossRefGoogle Scholar
  8. 8.
    Compant S, Clément C, Sessitsch A (2010) Plant growth-promoting bacteria in the rhizo- and endosphere of plants. Their role, colonization, mechanisms involved and prospects for utilization. Soil Biol Biochem 42:669–678CrossRefGoogle Scholar
  9. 9.
    Compant S, Duffy B, Nowak J, Clément C, Ait Barka E (2005) Use of plant growth-promoting bacteria for biocontrol of plant diseases: principles, mechanisms of action, and future prospects. Appl Environ Microbiol 71:4951–4959PubMedCrossRefGoogle Scholar
  10. 10.
    Compant S, Kaplan H, Sessitsch A, Nowak J, Ait Barka E, Clément C (2008) Endophytic colonization of Vitis vinifera L. by Burkholderia phytofirmans strain PsJN: from the rhizosphere to inflorescence tissues. FEMS Microbiol Ecol 63:84–93PubMedCrossRefGoogle Scholar
  11. 11.
    Compant S, Reiter B, Sessitsch A, Nowak J, Clément C, Ait Barka E (2005) Endophytic colonization of Vitis vinifera L. by plant growth-promoting bacterium Burkholderia sp. strain PsJN. Appl Environ Microbiol 71:1685–1693PubMedCrossRefGoogle Scholar
  12. 12.
    Daims H, Brühl A, Amann R, Schleifer K-H, Wagner M (1999) The domain-specific probe EUB338 is insufficient for the detection of all Bacteria: Development and evaluation of a more comprehensive probe set. Syst Appl Microbiol 22:434–444PubMedGoogle Scholar
  13. 13.
    Edwards U, Rogall T, Blocker H, Emde M, Bottger EC (1989) Isolation and direct complete nucleotide determination of entire genes. Characterization of a gene coding for 16S ribosomal RNA. Nucleic Acids Res 17:7843–7853PubMedCrossRefGoogle Scholar
  14. 14.
    Felske A, Akkermans ADL, De Vos WM (1998) In situ detection of an uncultured predominant Bacillus in Dutch grassland soils. Appl Environ Microbiol 64:4588–4590PubMedGoogle Scholar
  15. 15.
    Gerrath JM (1993) Developmental morphology and anatomy of grape flowers. Hortic Rev 13:315–337Google Scholar
  16. 16.
    Hallmann J (2001) Plant interactions with endophytic bacteria. In: Jeger MJ, Spence NJ (eds) Biotic interactions in plant–pathogen associations. CABI, Wallingford, pp 87–119CrossRefGoogle Scholar
  17. 17.
    Hallmann J, Berg B (2007) Spectrum and population dynamics of bacterial root endophytes. In: Schulz BJE, Boyle CJC, Sieber TN (eds) Microbial root endophytes. Springer, Berlin, pp 15–31Google Scholar
  18. 18.
    Hardoim PR, van Overbeek LS, Elsas JD (2008) Properties of bacterial endophytes and their proposed role in plant growth. Trends Microbiol 16:463–471PubMedCrossRefGoogle Scholar
  19. 19.
    Küsel K, Pinkart HC, Drake HL, Devereux R (1999) Acetogenic and sulfate-reducing bacteria inhabiting the rhizoplane and deep cortex cells of the sea grass Halodule wrightii. Appl Environ Microbiol 65:5117–5123PubMedGoogle Scholar
  20. 20.
    Liu WT, Marsh TL, Cheng H, Forney LJ (1997) Characterization of microbial diversity by determining terminal restriction fragment length polymorphisms of genes encoding 16S rRNA. Appl Environ Microbiol 63:4516–4522PubMedGoogle Scholar
  21. 21.
    López-López A, Rogel MA, Ormeño-Orrillo E, Martínez-Romero J, Martínez-Romero E (2010) Phaseolus vulgaris seed-borne endophytic community with novel bacterial species such as Rhizobium endophyticum sp. nov. Syst Appl Microbiol 33:322–327PubMedCrossRefGoogle Scholar
  22. 22.
    Loy A, Maixner F, Wagner M, Horn M (2007) probeBase—an online resource for rRNA-targeted oligonucleotide probes: new features 2007. Nucleic Acids Res 35:800–804CrossRefGoogle Scholar
  23. 23.
    Lugtenberg B, Kamilova F (2009) Plant-growth-promoting rhizobacteria. Annu Rev Microbiol 63:541–556PubMedCrossRefGoogle Scholar
  24. 24.
    Massol-Deya AA, Odelson DA, Hickey RF, Tiedje JM (1995) Bacterial community fingerprinting of amplified 16S and 16S-23S ribosomal DNA genes sequences and restriction endonuclease analysis (ARDRA). In: Akkermans ADL, van Elsas JD, de Bruijn FJ (eds) Molecular microbial ecology manual. Kluwer, Dordrecht, pp 1–18Google Scholar
  25. 25.
    Meier U (2001) Grapevine. Growth Stage of Mono- and Dicotyledonous Plants, BBCH Monograph. (Federal Biological Research Centre for Agriculture and Forestry, ed), Blackwell Wissenschafts, Berlin, pp. 91–93Google Scholar
  26. 26.
    Mundt JO, Hinkle NF (1976) Bacteria within ovules and seeds. Appl Environ Microbiol 32:694–698PubMedGoogle Scholar
  27. 27.
    Okunishi S, Sako K, Mano H, Imamura A, Morisaki H (2005) Bacterial flora of endophytes in the maturing seed of cultivated rice (Oryza sativa). Microbes Environ 20:168–177CrossRefGoogle Scholar
  28. 28.
    Rasche F, Hödl V, Poll C, Kandeler E, Gerzabek MH, van Elsas JD, Sessitsch A (2006) Rhizosphere bacteria affected by transgenic potatoes with antibacterial activities compared with the effects of soil, wild-type potatoes, vegetation stage and pathogen exposure. FEMS Microbiol Ecol 56:219–235PubMedCrossRefGoogle Scholar
  29. 29.
    Rosenblueth M, Martínez-Romero E (2006) Bacterial endophytes and their interaction with hosts. Mol Plant-Microbe Interact 19:827–837PubMedCrossRefGoogle Scholar
  30. 30.
    Weisburg WG, Barns SM, Pelletier DA, Lane DJ (1991) 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol 173:697–703PubMedGoogle Scholar
  31. 31.
    Welbaum G, Sturz AV, Dong Z, Nowak J (2004) Fertilizing soil microorganisms to improve productivity of agroecosystems. Crit Rev Plant Sci 23:175–193CrossRefGoogle Scholar
  32. 32.
    West ER, Cother EJ, Steel CC, Ash GJ (2010) The characterization and diversity of endophytes of grapevine. Can J Microbiol 56:209–216PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Stéphane Compant
    • 1
    • 2
    • 3
    Email author
  • Birgit Mitter
    • 1
  • Juan Gualberto Colli-Mull
    • 1
    • 4
  • Helmut Gangl
    • 5
  • Angela Sessitsch
    • 1
  1. 1.Bioresources UnitAIT Austrian Institute of Technology GmbHSeibersdorfAustria
  2. 2.Départment Bioprocédés et Systèmes Microbiens, ENSAT-INP de ToulouseUniversité de Toulouse, LGC UMR 5503 (CNRS/INPT/UPS)Castanet-Tolosan Cedex 1France
  3. 3.Centre de Viticulture-Œnologie de Midi PyrénéesUniversité de ToulouseCastanet-Tolosan Cedex 1France
  4. 4.Cinvestav-U. IrapuatoIrapuatoMexico
  5. 5.Bundesamt für WeinbauEisenstadtAustria

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