Advertisement

Microbial Ecology

, Volume 70, Issue 4, pp 904–911 | Cite as

High-Throughput Sequencing Reveals Drastic Changes in Fungal Communities in the Phyllosphere of Norway Spruce (Picea abies) Following Invasion of the Spruce Bud Scale (Physokermes piceae)

  • Audrius MenkisEmail author
  • Adas Marčiulynas
  • Artūras Gedminas
  • Jūratė Lynikienė
  • Aistė Povilaitienė
Fungal Microbiology

Abstract

The aim of this study was to assess the diversity and composition of fungal communities in damaged and undamaged shoots of Norway spruce (Picea abies) following recent invasion of the spruce bud scale (Physokermes piceae) in Lithuania. Sampling was done in July 2013 and included 50 random lateral shoots from ten random trees in each of five visually undamaged and five damaged 40–50-year-old pure stands of P. abies. DNA was isolated from 500 individual shoots, subjected to amplification of the internal transcribed spacer of fungal ribosomal DNA (ITS rDNA), barcoded and sequenced. Clustering of 149,426 high-quality sequences resulted in 1193 non-singleton contigs of which 1039 (87.1 %) were fungal. In total, there were 893 fungal taxa in damaged shoots and 608 taxa in undamaged shoots (p < 0.0001). Furthermore, 431 (41.5 %) fungal taxa were exclusively in damaged shoots, 146 (14.0 %) were exclusively in undamaged shoots, and 462 (44.5 %) were common to both types of samples. Correspondence analysis showed that study sites representing damaged and undamaged shoots were separated from each other, indicating that in these fungal communities, these were largely different and, therefore, heavily affected by P. piceae. In conclusion, the results demonstrated that invasive alien tree pests may have a profound effect on fungal mycobiota associated with the phyllosphere of P. abies, and therefore, in addition to their direct negative effect owing physical damage of the tissue, they may also indirectly determine health, sustainability and, ultimately, distribution of the forest tree species.

Keywords

Forest health Pathogens Pest insects Climate change Fungal community 

Notes

Acknowledgments

This research was funded by the European Regional Development Fund under the Global Grant measure, project no. VP1-3.1-ŠMM-07-K-02-001.

Conflict of Interest

All the authors declare no conflicts of interests.

Compliance with Ethical Standards

The authors declare the compliance of this work with ethical standards. All the authors are informed and agreed on the content of this work.

Supplementary material

248_2015_638_MOESM1_ESM.xlsx (174 kb)
ESM 1 (XLSX 173 kb)

References

  1. 1.
    Arias MMD, Batzer JC, Harrington TC, Wong AW, Bost SC, Cooley DR, Ellis MA, Hartman JR, Rosenberger DA, Sundin GW, Sutton TB, Travis JW, Wheeler MJ, Yoder KS, Gleason ML (2010) Diversity and biogeography of sooty blotch and flyspeck fungi on apple in the eastern and midwestern United States. Phytopathol 100:345–355CrossRefGoogle Scholar
  2. 2.
    Ben-Dov Y, Hodgson CJ (1997) Soft scale insects: their biology, natural enemies, and control, vol 1. Elsevier, The NetherlandsGoogle Scholar
  3. 3.
    Bradshaw RHW, Holmqvist BH, Cowling SA, Sykes MT (2000) The effects of climate change on the distribution and management of Picea abies in southern Scandinavia. Can J For Res 30:1992–1998CrossRefGoogle Scholar
  4. 4.
    Buee M, Reich M, Murat C, Morin E, Nilsson RH, Uroz S, Martin F (2009) 454 pyrosequencing analyses of forest soils reveal an unexpectedly high fungal diversity. New Phytol 184:449–456CrossRefPubMedGoogle Scholar
  5. 5.
    Chalmers N, Parker P (1989) Fieldwork and statistics for ecological projects, 2nd edn. The Open University, DorchesterGoogle Scholar
  6. 6.
    Davydenko K, Vasaitis R, Meshkova V, Menkis A (2014) Fungi associated with the red-haired bark beetle, Hylurgus ligniperda (Coleoptera: Curculionidae) in the forest-steppe zone in eastern Ukraine. Eur J Entomol 111:561–565Google Scholar
  7. 7.
    Fowler J, Cohen L, Jarvis P (1998) Practical statistics for field biology, 2nd edn. Wiley, ChichesterGoogle Scholar
  8. 8.
    Gedminas A, Lynikienė J, Marčiulynas A, Bagdžiūnaitė A (2015) Effect of Physokermes piceae Schrank. on shoot and needle growth in Norway spruce stands in Lithuania. Balt For 21:162–169Google Scholar
  9. 9.
    Graora D, Spasić R, Mihajlović L (2012) Bionomy of spruce bud scale, Physokermes piceae (Schrank.) (Hemiptera: Coccidae) in the Belgrade area, Serbia. Arch Biol Sci 64:337–343CrossRefGoogle Scholar
  10. 10.
    Hawksworth DL (2001) The magnitude of fungal diversity: the 1.5 million species estimate revisited. Mycol Res 105:1422–1431CrossRefGoogle Scholar
  11. 11.
    Hawksworth DL (2012) Global species numbers of fungi: are tropical studies and molecular approaches contributing to a more robust estimate? Biodivers Conserv 21:2425–2433CrossRefGoogle Scholar
  12. 12.
    Hibbett DS, Taylor JW (2013) Fungal systematics: is a new age of enlightenment at hand? Nature Rev Microbiol 11:129–133CrossRefGoogle Scholar
  13. 13.
    Ihrmark K, Bodeker ITM, Cruz-Martinez K, Friberg H, Kubartova A, Schenck J, Strid Y, Stenlid J, Brandstrom-Durling M, Clemmensen KE, Lindahl BD (2012) New primers to amplify the fungal ITS2 region—evaluation by 454-sequencing of artificial and natural communities. Fems Microbiol Ecol 82:666–677CrossRefPubMedGoogle Scholar
  14. 14.
    Kolar J (2007) The harmful entomofauna of woody plants in Slovakia. Acta Entomol Serb 12:67–79Google Scholar
  15. 15.
    Kosztarab M, Kozar F (1978) Scale insects—Coccoidea. Fauna Hungariae, Akadémiai Kiadó, BudapestGoogle Scholar
  16. 16.
    Kozar F (1985) New data to the knowledge of scale-insects of Bulgaria, Greece and Rumania (Homoptera: Coccoidea). Phytopathol Entomol Hung 20:201–205Google Scholar
  17. 17.
    Lagowska B (1986) Scale insects (Homoptera, Coccinea) of Roztocze and the Lublin Upland. Bulletin Entomologique de Pologne 56:475–478Google Scholar
  18. 18.
    Livsey S (1995) Ecology of endophytic microfungi in Norway spruce crowns. PhD thesis, Department of Forest Mycology and Pathology, Swedish University of Agricultural Sciences, UppsalaGoogle Scholar
  19. 19.
    Magurran AE (1988) Ecological diversity and its measurement. Princeton University Press, PrincetonCrossRefGoogle Scholar
  20. 20.
    Mead R, Curnow RN (1983) Statistical methods in agriculture and experimental biology. Chapman & Hall, LondonCrossRefGoogle Scholar
  21. 21.
    Menkis A, Burokienė D, Gaitnieks T, Uotila A, Johannesson H, Rosling A, Finlay RD, Stenlid J, Vasaitis R (2012) Occurrence and impact of the root-rot biocontrol agent Phlebiopsis gigantea on soil fungal communities in Picea abies forests of northern Europe. Fems Microbiol Ecol 81:438–445CrossRefPubMedGoogle Scholar
  22. 22.
    Menkis A, Urbina H, James TY, Rosling A (2014) Archaeorhizomyces borealis sp. nov. and a sequence-based classification of related soil fungal species. Fungal Biol 118:943–955CrossRefPubMedGoogle Scholar
  23. 23.
    Miezite O, Okmanis M, Indriksons A, Ruba J, Polmanis K, Freimane L (2013) Assessment of sanitary conditions in stands of Norway spruce (Picea abies Karst.) damaged by spruce bud scale (Physokermes piceae Schrnk.). iForest 6:73–78CrossRefGoogle Scholar
  24. 24.
    Muhlenberg E, Stadler B (2005) Effects of altitude on aphid-mediated processes in the canopy of Norway spruce. Agric For Entomol 7:133–143CrossRefGoogle Scholar
  25. 25.
    Müller MM, Hallaksela AM (1998) Diversity of Norway spruce needle endophytes in various mixed and pure Norway spruce stands. Mycol Res 102:1183–1189CrossRefGoogle Scholar
  26. 26.
    Müller MM, Hallaksela AM (2000) Fungal diversity in Norway spruce: a case study. Mycol Res 104:1139–1145CrossRefGoogle Scholar
  27. 27.
    Persson Y, Vasaitis R, Langstrom B, Ohrn P, Ihrmark K, Stenlid J (2009) Fungi vectored by the bark beetle Ips typographus following hibernation under the bark of standing trees and in the forest litter. Microb Ecol 58:651–659CrossRefPubMedGoogle Scholar
  28. 28.
    Ploetz RC, Hulcr J, Wingfield MJ, de Beer ZW (2013) Destructive tree diseases associated with ambrosia and bark beetles: black swan events in tree pathology? Plant Dis 97:856–872CrossRefGoogle Scholar
  29. 29.
    Quail M, Smith M, Coupland P, Otto T, Harris S, Connor T, Bertoni A, Swerdlow H, Gu Y (2012) A tale of three next generation sequencing platforms: comparison of Ion Torrent, Pacific Biosciences and Illumina MiSeq sequencers. BMC Genomics 13:341PubMedCentralCrossRefPubMedGoogle Scholar
  30. 30.
    Ronaghi M, Uhlén M, Nyrén P (1998) A sequencing method based on real-time pyrophosphate. Science 281:363–365CrossRefPubMedGoogle Scholar
  31. 31.
    Santini A, Ghelardini L, De Pace C, Desprez-Loustau ML, Capretti P, Chandelier A, Cech T, Chira D, Diamandis S, Gaitniekis T, Hantula J, Holdenrieder O, Jankovsky L, Jung T, Jurc D, Kirisits T, Kunca A, Lygis V, Malecka M, Marcais B, Schmitz S, Schumacher J, Solheim H, Solla A, Szabo I, Tsopelas P, Vannini A, Vettraino AM, Webber J, Woodward S, Stenlid J (2013) Biogeographical patterns and determinants of invasion by forest pathogens in Europe. New Phytol 197:238–250CrossRefPubMedGoogle Scholar
  32. 32.
    Scattolin L, Montecchio L (2009) Lophodermium piceae and Rhizosphaera kalkhoffii in Norway spruce: correlations with host age and climatic features. Phytopathol Mediterr 48:226–239Google Scholar
  33. 33.
    Schlyter P, Stjernquist I, Barring L, Jonsson AM, Nilsson C (2006) Assessment of the impacts of climate change and weather extremes on boreal forests in northern Europe, focusing on Norway spruce. Clim Res 31:75–84CrossRefGoogle Scholar
  34. 34.
    Shannon CE (1948) A mathematical theory of communication. Bell Syst Tech J 27:379–423CrossRefGoogle Scholar
  35. 35.
    Stadler B, Müller T (1996) Aphid honeydew and its effect on the phyllosphere microflora of Picea abies (L) Karst. Oecologia 108:771–776CrossRefGoogle Scholar
  36. 36.
    Stenlid J, Oliva J, Boberg JB, Hopkins AJM (2011) Emerging diseases in European forest ecosystems and responses in society. Forests 2:486–504CrossRefGoogle Scholar
  37. 37.
    Sykes MT, Prentice IC (1996) Climate change, tree species distributions and forest dynamics: a case study in the mixed conifer/northern hardwoods zone of northern Europe. Clim Chang 34:161–177CrossRefGoogle Scholar
  38. 38.
    ter Braak CJF, Smilauer P (1998) Canoco reference manual and user’s guide to Canoco for Windows: software for canonical community ordination, Version 4. Microcomputer Power, IthacaGoogle Scholar
  39. 39.
    Webb T (1986) Is vegetation in equilibrium with climate? How to interpret late-Quaternary pollen data. Vegetatio 67:75–91CrossRefGoogle Scholar
  40. 40.
    White TJ, Bruns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds) PCR protocols: a guide to methods and applications. Academic, San Diego, pp 315–322Google Scholar
  41. 41.
    Zhang X (1998) A study on the taxonomy of Exobasidium spp. according to fuzzy analysis of cultural properties and the analysis of 28S rDNA-PCR-RFLP. Scientia Silvae Sinicae 34:59–71Google Scholar
  42. 42.
    Zhuang JL, Zhu MQ, Zhang R, Yin H, Lei YP, Sun GY, Gleason ML (2010) Phialophora sessilis, a species causing flyspeck signs on bamboo in China. Mycotaxon 113:405–413CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Audrius Menkis
    • 1
  • Adas Marčiulynas
    • 2
  • Artūras Gedminas
    • 2
  • Jūratė Lynikienė
    • 2
  • Aistė Povilaitienė
    • 2
  1. 1.Department of Forest Mycology and Plant Pathology, Uppsala BioCenterSwedish University of Agricultural SciencesUppsalaSweden
  2. 2.Institute of Forestry, Lithuanian Research Centre for Agriculture and ForestryKaunasLithuania

Personalised recommendations