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Endophytic fungi in European aspen (Populus tremula) leaves—diversity, detection, and a suggested correlation with herbivory resistance

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Abstract

Fungal endophytes are found in most seed plants, but their ecological function mainly remains elusive, except in pooid (or clavicipitalean) systems. The diversity and dynamics of endophytes in non clavicipitalean plants make studies of their ecological function challenging. This paper describes the advantage of using molecular techniques to survey the ecological function of endophytes in Populus tremula clones. About 1,000 endophyte isolates were distinguished using traditional methods; these isolates represented approximately 100 morphologically distinct groups. We generated 73 DNA-sequences (18S and ITS rDNA) from these groups and determined 33 distinct taxa. They represented the Basidiomycota and Ascomycota, including diverse Sordariomycetes and Dothideomycetes, and three sequences that were identified, by their ITS sequences, as a species of Epicoccum (a genus of supposedly unknown relation within the Ascomycota) were placed within the Pleosporales, in the 18S phylogeny. Primer pairs were designed for eleven of the fungi. Of these, three primers produced bands for a subset of Aspen samples. The primer pairs allowed endophytes in field samples to be readily identified, with a detection limit of 0.15 percent fungal DNA. The presence of fungi in Aspen clones was related to field damage by herbivores and the pathogen Venturia tremula. A negative association was found in two separate surveys between Aureobasidium sp. and herbivore damage, but we found no evidence that endophyte presence was related to a history of Venturia symptoms. This approach promises to enhance greatly the scope for qualitative and quantitative detection of endophyte communities, and to improve our ability to elucidate the ecological function of non clavicipitalean endophytes.

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References

  • Ahlholm J, Helander M, Elamo P, Saloniemi I, Neuvonen S, Hanhimäki S, Saikkonen K (2002) Micro-fungi and invertebrate herbivores on birch trees: fungal mediated plant-herbivore interactions or responses to host quality? Ecol Lett 5:648–655

    Article  Google Scholar 

  • Andersson A, Keskitalo J, Sjödin A, Bhalerao R, Sterky F, Wissel K, Tandre K, Aspeborg H, Moyle R, Ohmiya Y, Bhalerao R, Brunner A, Gustafsson P, Karlsson J, Lundeberg J, Nilsson O, Sandberg G, Strauss S, Sundberg B, Uhlen M, Jansson S, Nilsson P (2004) A transcriptional timetable of autumn senescence. Genome Biol 5(4):R24

    Article  PubMed  Google Scholar 

  • Arnold AE (2007) Understanding the diversity of foliar endophytic fungi: progress, challenges and frontiers. Fungal Biol Rev 21:51–66

    Article  Google Scholar 

  • Arnold AE, Maynard Z, Gilbert GS, Coley PD, Kursar TA (2000) Are tropical fungal endophytes hyperdiverse? Ecol Lett 3:267–274

    Article  Google Scholar 

  • Arnold AE, Mejia LC, Kyllo D, Enith I, Rojas EI, Maynard Z, Robbins N, Herre EA (2003) Fungal endophytes limit pathogen damage in a tropical tree. Proc Natl Acad Sci U S A 100:15649–15654

    Article  CAS  PubMed  Google Scholar 

  • Arnold AE, Henk DA, Eells RL, Lutzoni F, Vilgalys R (2007) Diversity and phylogenetic affinities of foliar fungal endophytes in loblolly pine inferred by culturing and environmental PCR. Mycologia 99:185–206

    Article  CAS  PubMed  Google Scholar 

  • Bailey JK, Deckert R, Schweitzer JA, Rehill BJ, Lindroth RL, Gehring C, Whitham TG (2005) Host plant genetics affect hidden ecological players, links among Populus, condensed tannins, and fungal endophyte infection. Can J Bot 83:356–361

    Article  Google Scholar 

  • Bazely DR, Vicari M, Emmerich S, Filip L, Lin D, Inman A (1997) Interactions between herbivores and endophyte-infected Festuca rubra from the Scottish Islands of St. Kilda, Benbecula and Rum. J Appl Ecol 34:847–860

    Article  Google Scholar 

  • Bencheqroun SK, Baji M, Massart S, Labhilili M, El Jaafari S, Jijakli MH (2007) In vitro and in situ study of postharvest apple blue mold biocontrol by Aureobasidium pullulans: evidence for the involvement of competition for nutrients. Postharvest Biol Technol 46:128–135

    Article  CAS  Google Scholar 

  • Carroll G (1995) Forest endophytes—pattern and process. Can J Bot 73(S1):S1316–S1324

    Article  Google Scholar 

  • Clay K (1988) Fungal endophytes of grasses: a defensive mutualism between plants and fungi. Ecology 69:10–16

    Article  Google Scholar 

  • Curlevski NJA, Xu ZH, Anderson IC, Cairney JWG (2010) Diversity of soil and rhizosphere fungi under Araucaria bidwillii (Bunya pine) at an Australian tropical montane rainforest site. Fungal Divers 40. doi:10.1007/s13225-009-0001-0.

  • Devarajan PT, Suryanarayanan TS (2006) Evidence for the role of phytophagous insects in dispersal of non-grass fungal endophytes. Fungal Divers 23:111–119

    Google Scholar 

  • Duong LM, Jeewon R, Lumyong S, Hyde KD (2006) DGGE coupled with ribosomal DNA gene phylogenies reveal uncharacterized fungal phylotypes. Fungal Divers 23:121–138

    Google Scholar 

  • Elamo P, Helander ML, Saloniemi I, Neuvonen S (1999) Birch family and environmental conditions affect endophytic fungi in leaves. Oecologia 118:151–156

    Article  Google Scholar 

  • Farris JS, Albert VA, Källersjö M, Lipscomb D, Kluge AG (1996) Parsimony jackknifing outperforms neighbor-joining. Cladistics 12:99–124

    Article  Google Scholar 

  • Faeth SH (2002) Are endophytic fungi defensice plant mutualists? Oikos 98:25–36

    Article  Google Scholar 

  • Faeth SH, Hammon KE (1997) Fungal endophytes in oak trees; long term patterns of abundance and associations with leafminers. Ecology 78:810–819

    Article  Google Scholar 

  • Ganley RJ, Brunsfeld SJ, Newcombe G (2004) A community of unknown, endophytic fungi in western white pine. Proc Natl Acad Sci USA 101:10107–10112

    Article  CAS  PubMed  Google Scholar 

  • Gargas A, DePriest PT (1996) A nomenclature for fungal PCR primers with examples from intron-containing SSU rDNA. Mycologia 88:745–748

    Article  CAS  Google Scholar 

  • Giordano L, Gonthier P, Varese GC, Miserere L, Nicolotti G (2009) Mycobiota inhabiting sapwood of healthy and declining Scots pine (Pinus sylvestris L.) trees in the Alps. Fungal Divers 38:69–83

    Google Scholar 

  • Gonthier P, Gennaro M, Nicolotti G (2006) Effects of water stress on the endophytic mycota of Quercus robur. Fungal Divers 21:69–80

    Google Scholar 

  • Granath G, Vicari M, Bazely DR, Ball JP, Puentes A, Rakocevic T (2007) Variation in the abundance of fungal endophytes in fescue grasses along altitudinal and grazing gradients. Ecography 30:422–430

    Google Scholar 

  • Grass Phylogeny Working Group (2001) Phylogeny and subfamilial classification of the grasses (Poaceae). Ann Mo Bot Gard 88:373–457

    Article  Google Scholar 

  • Guo LD, Huang GR, Wang Y, He WH, Zheng WH, Hyde KD (2003) Molecular identification of white morphotype strains of endophytic fungi from Pinus tabulaeformis. Mycol Res 107:680–688

    Article  CAS  PubMed  Google Scholar 

  • Hatcher MJ, Dick JTA, Dunn AM (2006) How parasites affect interactions between competitors and predators. Ecol Lett 9:1253–1271

    Article  PubMed  Google Scholar 

  • Hibbett DS et al (2007) A higher-level phylogenetic classification of the fungi. Mycol Res 111:509–548

    Article  PubMed  Google Scholar 

  • Hu HL, Jeewon R, Zhou DQ, Zhou TX, Hyde KD (2007) Phylogenetic diversity of endophytic Pestalotiopsis species in Pinus armandii and Ribes spp.: evidence from rDNA and β-tubulin gene phylogenies. Fungal Divers 24:1–22

    CAS  Google Scholar 

  • Huang WY, Cai YZ, Hyde KD, Corke H, Sun M (2008) Biodiversity of endophytic fungi associated with 29 traditional Chinese medicinal plants. Fungal Divers 33:61–75

    Google Scholar 

  • Huang WY, Cai YZ, Surveswaran S, Hyde KD, Corke H, Sun M (2009) Molecular phylogenetic identification of endophytic fungi isolated from three Artemisia species. Fungal Divers 36:69–88

    CAS  Google Scholar 

  • Hyde KD, Soytong K (2008) The fungal endophyte dilemma. Fungal Divers 33:163–173

    Google Scholar 

  • Ingvarsson PK (2005) Nucleotide polymorphism and linkage disequilibrium within and among natural populations of European aspen (Populus tremula L., Salicaceae). Genetics 169:945–953

    Article  CAS  PubMed  Google Scholar 

  • Koh S, Hik DS (2007) Herbivory mediates grass-endophyte relationships. Ecology 88:2752–2757

    Article  PubMed  Google Scholar 

  • Krohn K, Biele C, Drogies KH, Steingrover K, Aust HJ, Draeger S, Schulz B (2002) Fusidilactones, a new group of polycyclic lactones from an endophyte, Fusidium sp. Eur J Org Chem 14:2331–2336

    Article  Google Scholar 

  • Kumar DSS, Hyde KD (2004) Biodiversity and tissue-recurrence of endophytic fungi from Tripterygium wilfordii. Fungal Divers 17:69–90

    CAS  Google Scholar 

  • Lacap DC, Hyde KD, Liew ECY (2003) An evaluation of the fungal ‘morphotype’ concept based on ribosomal DNA sequences. Fungal Divers 12:53–66

    Google Scholar 

  • Lappalainen JH, Koricheva J, Helander ML, Haukioja E (1999) Densities of endophytic fungi and performance of leafminers (Lepidoptera, Eriocraniidae) on birch along a pollution gradient. Environ Pollut 104:99–105

    Article  CAS  Google Scholar 

  • Li WC, Zhou J, Guo SY, Guo LD (2007) Endophytic fungi associated with lichens in baihua mountain of Beijing, China. Fungal Divers 25:69–80

    Google Scholar 

  • Luquez V, Hall D, Albrectsen BR, Karlsson J, Ingvarsson P, Jansson S (2008) Natural phenological variation in aspen (Populus tremula): the SwAsp collection. Tree Genet Genomes 4:279–292

    Article  Google Scholar 

  • Mitchell AM, Strobel GA, Hess WM, Vargas PN, Ezra D (2008) Muscodor crispans, a novel endophyte from Ananas ananassoides in the Bolivian Amazon. Fungal Divers 31:37–43

    Google Scholar 

  • Osier TL, Lindroth RL (2001) Effects of genotype, nutrient availability, and defoliation on aspen phytochemistry and insect performance. J Chem Ecol 27:1289–1313

    Article  CAS  PubMed  Google Scholar 

  • Osono T (2008) Endophytic and epiphytic phyllosphere fungi of Camelia japonica: sasonal and leaf age-dependent variations. Mycologia 100:387–391

    Article  PubMed  Google Scholar 

  • Petrini O (1991) Endophytes of tree leaves. In: Andrews JH, Hirano SS (eds) Microbial ecology of leaves. Brock/Springer Series in Contemporary Bioscience, pp 179–196

  • Petrini O, Fisher PJ (1990) Occurrence of fungal endophytes in twigs of Salix fragilis and Quercus robur. Mycol Res 94:1077–1080

    Article  Google Scholar 

  • Preszler RW, Gaylord ES, Boecklen WJ (1996) Reduced parasitism of a leaf mining moth on trees with high infection frequencies of an endophytic fungus. Oecologia 108:159–166

    Article  Google Scholar 

  • Rudger JA, Clay K (2008) An invasive plant-fungal mutualism reduces arthropod diversity. Ecol Lett 11:831–840

    Article  Google Scholar 

  • Rudger JA, Koslow JM, Clay K (2004) Endophytic fungi alter relationships between diversity and ecosystem properties. Ecol Lett 7:42–51

    Article  Google Scholar 

  • Rungjindamai N, Pinruan U, Choeyklin R, Hattori T, Jones EBG (2008) Molecular characterization of basidiomycetous endophytes isolated from leaves, rachis and petioles of the oil palm, Elaeis guineensis, in Thailand. Fungal Divers 33:139–162

    Google Scholar 

  • Rozen S, Skaletsky H (2000) Primer3 on the WWW for general users and for biologist programmers. In: Krawetz S, Misener S (eds) Bioinformatics methods and protocols in the series methods in molecular biology. Humana Press, Totowa, pp 365–386, http://primer3.sourceforge.net/

    Google Scholar 

  • Saikkonen K (2007) Forest structure and fungal endophytes. Fungal Biol Rev 21:67–74

    Article  Google Scholar 

  • Saikkonen K, Helander M, Ranta H, Neuvonen S, Virtanen T, Suomela L, Vuorinen P (1996) Endophyte mediated interactions between woody plants and insect herbivores. J Entomol Exp Appl 80:269–271

    Article  Google Scholar 

  • Saikkonen K, Faeth SH, Helander M, Sullivan TJ (1998) Fungal endophytes: a continuum of interactions with host plants. Annu Rev Ecol Systemat 29:319–343

    Article  Google Scholar 

  • Sánchez Márquez S, Bills GF, Zabalgogeazcoa I (2008) Diversity and structure of the fungal endophytic assemblages from two sympatric coastal grasses. Fungal Divers 33:87–100

    Google Scholar 

  • Santamaria O, Diez JJ (2005) Fungi in leaves, twigs and stem bark of Populus tremula from northern Spain. Forest Pathol 35:95–104

    Article  Google Scholar 

  • Saona NM, Albrectsen BR, Ericson L, Bazely DR (2010) Environmental stresses mediate grass-endophyte interactions in a boreal archipelago. J Ecol. doi:10.1111/j.1365-2745.2009.01613.x

  • Schulz B, Boyle C (2005) The endophytic continuum. Mycol Res 109:661–686

    Article  PubMed  Google Scholar 

  • Schulz B, Boyle C, Draeger S, Rmmert AK, Krohn K (2002) Endophytic fungi: a source of novel biologically active secondary metabolites. Mycol Res 106:996–1004

    Article  CAS  Google Scholar 

  • Schweitzer JA, Madritch MD, Bailey JK, LeRoy CJ, Fischer DG, Rehill BJ, Lindroth RL, Hagerman AE, Wooley SC, Hart SC, Whitham TG (2008) From genes to ecosystems: the genetic basis of condenced tannins and their role in nutrient regulation in a Populus model system. Ecosystems 11:1005–1020

    Article  CAS  Google Scholar 

  • Seena S, Wynberg N, Bärlocher F (2008) Fungal diversity during leaf decomposition in a stream assessed through clone libraries. Fungal Divers 30:1–14

    Google Scholar 

  • Segerman B, Jansson S, Karlsson J (2007) Characterization of genes with tissue-specific differential expression patterns in Populus. Tree Genet Genomes 3:351–362

    Article  Google Scholar 

  • Service RF (2006) The race for the 1000$ genome. Science 311:1544–1546

    Article  CAS  PubMed  Google Scholar 

  • Sieber TN (2007) Endophytic fungi in forest trees: are they mutualists? Fungal Biol Rev 21:75–89

    Article  Google Scholar 

  • Sjödin A, Bylesjö M, Skogström O, Eriksson D, Nilsson P, Rydén P, Jansson S, Karlsson J (2006) UPSC-BASE—Populus transcriptomics online. Plant J 48:806–817

    Article  PubMed  CAS  Google Scholar 

  • Sjödin A, Wissel K, Bylesjö M, Trygg J, Jansson S (2008) Global expression profiling in leaves of free-growing aspen. Biomed Central Plant Biol 8:61

    Google Scholar 

  • Skrzyńska M (2004) Studies on population frequency of insects and mites causing galls on the leaves of European aspen Populus tremula L. in southern Poland. J Pest Sci 77:119–122

    Article  Google Scholar 

  • Stierle A, Strobel G, Stierle D (1993) Taxol and taxane production by Taxomyces andreanae, an endophytic fungus of Pacific yew. Science 260(5105):214–216

    Article  CAS  PubMed  Google Scholar 

  • Strobel G, Daisy B, Castillo U, Harper J (2004) Natural products from endophytic microorganisms. J Nat Products 67:257–268

    Article  CAS  Google Scholar 

  • Suryanarayanan TS, Venkatesan G, Murali TS (2003) Endophytic fungal communities in leaves of tropical forest trees: diversity and distribution patterns. Curr Sci 85:489–493

    Google Scholar 

  • Swofford DL (2002) PAUP* Phylogenetic analysis using parsimony (* and other methods), version 4.0b10. Sinauer Associates, Sunderland, Massachusetts, USA

  • Tao G, Liu ZY, Hyde KD, Lui XZ, Yu ZN (2008) Whole rDNA analysis reveals novel and endophytic fungi in Bletilla ochracea (Orchidaceae). Fungal Divers 33:101–122

    Google Scholar 

  • Tejesvi MV, Kini KR, Prakash HS, ven Subbiah, Shetty HS (2007) Genetic diversity and antifungal activity of species of Pestalotiopsis isolated as endophytes from medicinal plants. Fungal Divers 24:37–54

    Google Scholar 

  • Tejesvi MV, Tamhankar SA, Kini KR, Rao VS, Prakash HS (2009) Phylogenetic analysis of endophytic Pestalotiopsis species from ethnopharmaceutically important medicinal trees. Fungal Divers 38:167–183

    Google Scholar 

  • Tsai CJ, Harding SA, Tschaplinski TJ, Lindroth RL, Yuan YN (2006) Genome-wide analysis of the structural genes regulating defense phenylpropanoid metabolism in populus. New Phytol 172:47–62

    Article  CAS  PubMed  Google Scholar 

  • Tuskan GA et al (2006) The genome of black cottonwood, Populus tricocarpa (Torr. and Gray). Science 313:1596–1604

    Article  CAS  PubMed  Google Scholar 

  • Waller F, Achatz B, Baltruschat H, Fodor J, Becker K, Fischer M, Heier T, Huckelhoven R, Neumann C, von Wettstein D, Franken P, Kogel KH (2005) The endophytic fungus Piriformospora indica reprograms barley to salt-stress tolerance, disease resistance, and higher yield. Proc Natl Acad Sci U S A 102:13386–13391

    Article  CAS  PubMed  Google Scholar 

  • Wang Y, Guo LD, Hyde KD (2005) Taxonomic placement of sterile morphotypes of endophytic fungi from Pinus tabulaeformis (Pinaceae) in northeast China based on rDNA sequences. Fungal Divers 20:235–260

    CAS  Google Scholar 

  • Webber JF (1981) A natural biological control of Dutch elm disease. Nature 292:449–451

    Article  Google Scholar 

  • Wedin M, Döring H, Gilenstam G (2006) Stictis s. lat. (Ostropales, Ascomycota) in northern Scandinavia, with a key and notes on morphological variation in relation to lifestyle. Mycol Res 110:773–789

    Article  PubMed  Google Scholar 

  • White TJ, Bruns T, Lee S, Taylor JW (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 315–322

    Google Scholar 

  • Wilson D (1995) Endophyte—the evolution of a term, and clarification of its use and definition. Oikos 73:274–276

    Article  Google Scholar 

  • Wilson D, Carroll GC (1997) Avoidance of high-endophyte space by gall-forming insects. Ecology 78:2153–2163

    Article  Google Scholar 

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Acknowledgements

This study was inspired by a workshop held by Dr. Mark Vicari in Umeå in 2005 financed by the Edlunds Brothers foundation to BRA. Financial support was received from the Swedish Research Council, the Swedish Research Council for Environment, Agricultural Sciences and Spatial Planning (MW and SJ) and the Kempe foundation (BRA and SJ).

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Authors and Affiliations

  1. Department of Plant Physiology, Umeå University, Umeå Plant Science Centre, SE-901 87, Umeå, Sweden

    Benedicte R. Albrectsen, Lars Björkén, Akkamahadevi Varad, Jan Karlsson & Stefan Jansson

  2. Department of Ecology and Environmental Science, Umeå University, SE-901 87, Umeå, Sweden

    Åsa Hagner

  3. Department of Forest Resource Management, Swedish University of Agricultural Sciences, SE-901 87, Umeå, Sweden

    Åsa Hagner

  4. Cryptogamic Botany, The Swedish Museum of Natural History, Box 50007, SE-104 05, Stockholm, Sweden

    Mats Wedin

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  1. Benedicte R. Albrectsen
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  4. Åsa Hagner
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Correspondence to Benedicte R. Albrectsen.

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Suppl. Table 2

Summary of endophyte names for isolates in aspen 14/1 2008 (classification according to AFTOL) (DOC 42 kb)

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Albrectsen, B.R., Björkén, L., Varad, A. et al. Endophytic fungi in European aspen (Populus tremula) leaves—diversity, detection, and a suggested correlation with herbivory resistance. Fungal Diversity 41, 17–28 (2010). https://doi.org/10.1007/s13225-009-0011-y

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