Microbial Ecology

, Volume 64, Issue 3, pp 714–724 | Cite as

Exploring Biodiversity in the Bacterial Community of the Mediterranean Phyllosphere and its Relationship with Airborne Bacteria

  • Despoina VokouEmail author
  • Katerina Vareli
  • Ekaterini Zarali
  • Katerina Karamanoli
  • Helen-Isis A. Constantinidou
  • Nikolaos Monokrousos
  • John M. Halley
  • Ioannis Sainis
Plant Microbe Interactions


We studied the structure and diversity of the phyllosphere bacterial community of a Mediterranean ecosystem, in summer, the most stressful season in this environment. To this aim, we selected nine dominant perennial species, namely Arbutus unedo, Cistus incanus, Lavandula stoechas, Myrtus communis, Phillyrea latifolia, Pistacia lentiscus, Quercus coccifera (woody), Calamintha nepeta, and Melissa officinalis (herbaceous). We also examined the extent to which airborne bacteria resemble the epiphytic ones. Genotype composition of the leaf and airborne bacteria was analysed by using denaturing gradient gel electrophoresis profiling of a 16S rDNA gene fragment; 75 bands were cloned and sequenced corresponding to 28 taxa. Of these, two were found both in the air and the phyllosphere, eight only in the air, and the remaining 18 only in the phyllosphere. Only four taxa were found on leaves of all nine plant species. Cluster analysis showed highest similarity for the five evergreen sclerophyllous species. Aromatic plants were not grouped all together: the representatives of Lamiaceae, bearing both glandular and non-glandular trichomes, formed a separate group, whereas the aromatic and evergreen sclerophyllous M. communis was grouped with the other species of the same habit. The epiphytic communities that were the richest in bacterial taxa were those of C. nepeta and M. officinalis (Lamiaceae). Our results highlight the remarkable presence of lactic acid bacteria in the phyllosphere under the harsh conditions of the Mediterranean summer, the profound dissimilarity in the structure of bacterial communities in phyllosphere and air, and the remarkable differences of leaf microbial communities on neighbouring plants subjected to similar microbial inocula; they also point to the importance of the leaf glandular trichome in determining colonization patterns.


Bacterial Community Colony Form Unit Aromatic Plant Bacterial Taxon Natamycin 
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.



We thank Prof. S. E. Lindow, University of California, Berkeley, for his constructive comments on an earlier version of our manuscript.


  1. 1.
    Andrews JH (1992) Biological control in the phyllosphere. Annu Rev Phytopathol 30:603–635PubMedCrossRefGoogle Scholar
  2. 2.
    Baldotto LEB, Olivares FL (2008) Phyloepiphytic interaction between bacteria and different plant species in a tropical agricultural system. Can J Microbiol 54:918–931PubMedCrossRefGoogle Scholar
  3. 3.
    Bowers RM, McLetchie S, Knight R, Fierer N (2011) Spatial variability in airborne bacterial communities across land-use types and their relationship to the bacterial communities of potential source environments. ISME J 5:601–612PubMedCrossRefGoogle Scholar
  4. 4.
    Chalkos DG (2010) Effects of secondary metabolites of aromatic plants on the structure and metabolism of the soil microbial communities. PhD thesis. Aristotle University of Thessaloniki: Thessaloniki, Greece (in Greek with English summary)Google Scholar
  5. 5.
    Constantinidou ΗΑ, Hirano SS, Baker LS, Upper CD (1990) Atmospheric dispersal of ice nucleation-active bacteria: the role of rain. Phytopathology 80:934–937CrossRefGoogle Scholar
  6. 6.
    de Jager ES, Wehner FC, Korsten L (2001) Microbial ecology of the mango phylloplane. Microb Ecol 42:201–207PubMedGoogle Scholar
  7. 7.
    Delmotte N, Knief C, Chaffron S, Innerebner G, Roschitzki B, Schlapbach R, von Meringb C, Vorholta JA (2009) Community proteogenomics reveals insights into the physiology of phyllosphere bacteria. Proc Natl Acad Sci USA 106:16428–16433PubMedCrossRefGoogle Scholar
  8. 8.
    Fierer N, McCain CM, Meir P, Zimmermann M, Rapp JM, Silman MR, Knight R (2011) Microbes do not follow the elevation diversity patterns of plants and animals. Ecology 92:797–804PubMedCrossRefGoogle Scholar
  9. 9.
    Karamanoli K, Vokou D, Menkissoglu O, Constantinidou HΙ (2000) Bacterial colonization of the phyllosphere of Mediterranean aromatic plants. J Chem Ecol 26:2035–2048CrossRefGoogle Scholar
  10. 10.
    Knief C, Ramette A, Frances L, Alonso-Blanco C, Vorholt JA (2010) Site and plant species are important determinants of the Methylobacterium community composition in the plant phyllosphere. ISME J 4:719–728PubMedCrossRefGoogle Scholar
  11. 11.
    Lambais MR, Crowley DE, Cury JC, Bull RC, Rodrigues RR (2006) Bacterial diversity in tree canopies of the Atlantic forest. Science 312:1917–1917PubMedCrossRefGoogle Scholar
  12. 12.
    Lavermicocca P, Gobbetti M, Corsetti A, Caputo L (1998) Characterization of lactic acid bacteria isolated from olive phylloplane and table olive brines. Ital J Food Sci 10:27–39Google Scholar
  13. 13.
    Lindemann J, Constantinidou HΑ, Barchet WR, Upper CD (1982) Plants as sources of airborne bacteria, including ice nucleation-active bacteria. Appl Environ Microbiol 44:1059–1063PubMedGoogle Scholar
  14. 14.
    Lindow SE (1996) Role of immigration and other processes in determining epiphytic bacterial populations: implications for disease management. In: Morris CE, Nicot PC, Nguyen C (eds) Aerial plant surface microbiology. Plenum, New York, pp 155–168CrossRefGoogle Scholar
  15. 15.
    Mariano RLR, McCarter SM (1993) Epiphytic survival of Pseudomonas viridiflava on tomato and selected weed species. Microb Ecol 26:47–58CrossRefGoogle Scholar
  16. 16.
    Mew TW, Vera Cruz CM (1986) Epiphytic colonization of host and non-host plants by phytopathogenic bacteria. In: Fokkema NJ, van den Heuvel J (eds) Microbiology of the phyllosphere. Cambridge University Press, New York, pp 269–282Google Scholar
  17. 17.
    Muller T, Behrendt U, Muller M (1996) Antagonistic activity in plant-associated lactic acid bacteria. Microbiol Res 151:63–70CrossRefGoogle Scholar
  18. 18.
    Mundt JO, Coggins JH, Johnson LF (1962) Growth of Streptococcus faecalis var. liquefaciens on plants. Appl Microbiol 10:552–555PubMedGoogle Scholar
  19. 19.
    Muyzer G, Teske A, Wirsen CO, Jannasch HW (1995) Phylogenetic relationships of Thiomicrospira species and their identification in deep-sea hydrothermal vent samples by denaturing gradient gel electrophoresis of 16S rDNA fragments. Arch Microbiol 164:165–171PubMedCrossRefGoogle Scholar
  20. 20.
    Pugh GJF, Buckley NJ (1971) The leaf surface as substrate for colonization by fungi. In: Preece TF, Dickinson CH (eds) Ecology of leaf surface micro-organisms. Academic, London, pp 431–445Google Scholar
  21. 21.
    Quaiser A, Ochsenreiter T, Lanz C, Schuster SC, Treusch AH, Eck J, Schleper C (2003) Acidobacteria form a coherent but highly diverse group within the bacterial domain: evidence from environmental genomics. Mol Microbiol 50:563–575PubMedCrossRefGoogle Scholar
  22. 22.
    Redford AJ, Bowers RM, Knight R, Linhart Y, Fierer N (2010) The ecology of the phyllosphere: geographic and phylogenetic variability in the distribution of bacteria on tree leaves. Environ Microbiol 12:2885–2893PubMedCrossRefGoogle Scholar
  23. 23.
    Schönherr J, Baur P (1996) Cuticle permeability studies: a model for estimating leaching of plant metabolites to leaf surfaces. In: Morris CE, Nicot PC, Nguyen C (eds) Aerial plant surface microbiology. Plenum, New York, pp 1–23CrossRefGoogle Scholar
  24. 24.
    Sokal R, Michener C (1958) A statistical method for evaluating systematic relationships. Univ Kans Sci Bull 38:1409–1438Google Scholar
  25. 25.
    Vareli K, Briasoulis E, Pilidis G, Sainis I (2009) Molecular confirmation of Planktothrix rubescens as the cause of intense, microcystin-synthesizing cyanobacterial bloom in Lake Ziros, Greece. Harmful Algae 8:447–453CrossRefGoogle Scholar
  26. 26.
    Vokou D (2007) Allelochemicals, allelopathy and essential oils: a field in search of definitions and structure. Allelopathy J 19:119–135Google Scholar
  27. 27.
    Vokou D, Chalkos D, Karamanoli K (2006) Microorganisms and allelopathy: a one-sided approach. In: Reigosa MJ, Pedrol N, Gonzalez L (eds) Allelopathy: a physiological process with ecological implications. Springer, Dordrecht, pp 341–371Google Scholar
  28. 28.
    Vokou D, Margaris NS (1988) Decomposition of terpenes by soil microorganisms. Pedobiologia 31:413–419Google Scholar
  29. 29.
    Vokou D, Margaris NS, Lynch JM (1984) Effects of volatile oils from aromatic shrubs on soil microorganisms. Soil Biol Biochem 16:509–513CrossRefGoogle Scholar
  30. 30.
    Whipps JM, Hand P, Pink D, Bending GD (2008) Phyllosphere microbiology with special reference to diversity and plant genotype. J Appl Microbiol 105:1744–1755PubMedCrossRefGoogle Scholar
  31. 31.
    Yadav RKP, Bosabalidis AM, Vokou D (2004) Leaf structural features of Mediterranean perennial species: plasticity and life form specificity. J Biol Res 2:21–34Google Scholar
  32. 32.
    Yadav RKP, Halley JM, Karamanoli K, Constantinidou HI, Vokou D (2004) Bacterial populations on the leaves of Mediterranean plants: quantitative features and testing of distribution models. Environ Exp Bot 52:63–77CrossRefGoogle Scholar
  33. 33.
    Yadav RKP, Karamanoli K, Vokou D (2011) Bacterial populations on the phyllosphere of Mediterranean plants: influence of leaf age and leaf surface. Front Agr China 5:60–63CrossRefGoogle Scholar
  34. 34.
    Yadav RKP, Karamanoli K, Vokou D (2005) Bacterial colonization of the phyllosphere of Mediterranean perennial species as influenced by leaf structural and chemical features. Microb Ecol 50:185–196PubMedCrossRefGoogle Scholar
  35. 35.
    Yadav RKP, Papatheodorou EM, Karamanoli K, Constantinidou H-IA, Vokou D (2008) Abundance and diversity of the phyllosphere bacterial communities of Mediterranean perennial plants that differ in leaf chemistry. Chemoecology 18:217–226CrossRefGoogle Scholar
  36. 36.
    Yang CH, Crowley DE, Borneman J, Keen NT (2001) Microbial phyllosphere populations are more complex than previously realized. Proc Natl Acad Sci USA 98:3889–3894PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Despoina Vokou
    • 1
    Email author
  • Katerina Vareli
    • 2
  • Ekaterini Zarali
    • 3
  • Katerina Karamanoli
    • 4
  • Helen-Isis A. Constantinidou
    • 4
  • Nikolaos Monokrousos
    • 1
  • John M. Halley
    • 2
  • Ioannis Sainis
    • 5
  1. 1.Department of Ecology, School of BiologyAristotle UniversityThessalonikiGreece
  2. 2.Department of Biological Applications and TechnologyUniversity of IoanninaIoanninaGreece
  3. 3.Medical SchoolUniversity of IoanninaIoanninaGreece
  4. 4.School of AgricultureAristotle UniversityThessalonikiGreece
  5. 5.Interscience Molecular Oncology Laboratory, Human Cancer Biobank CenterUniversity of IoanninaIoanninaGreece

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