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Diversity, Molecular Phylogeny, and Bioactive Potential of Fungal Endophytes Associated with the Himalayan Blue Pine (Pinus wallichiana)

  • Plant Microbe Interactions
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Abstract

In this study, we investigated the diversity of fungal endophytes associated with Pinus wallichiana from the Western Himalayas, with emphasis on comparison of endophytic communities harbored by the stem and needle tissues of the host and their antimicrobial potential. A total number of 130 isolates, comprising of 38 different genera, were recovered from 210 fragments of the plant. Among the isolated fungi, only a single isolate, Tritirachium oryzae, belonged to the Phylum Basidiomycota whereas the rest belonged to Ascomycota. Dothideomycetes was the dominant class with the highest isolation frequency of 49.2 %. The most frequent colonizers of the host were Alternaria spp., Pestalotiopsis spp., Preussia spp., and Sclerostagonospora spp. The diversity and species richness were higher in needle tissues than in the stems. Antimicrobial activities were displayed by extracts from a total number of 22 endophytes against one or more pathogens. Endophytes designated as P1N13 (Coniothyrium carteri), P2N8 (Thielavia subthermophila), P4S6b (Truncatella betulae), P7N10 (Cochliobolus australiensis), and P8S4 (Tritirachium oryzae) were highly active against Candida albicans. Broad spectrum antimicrobial activities were obtained with the extracts of P8-S4 (Tritirachium oryzae) and P5-N26 (Coniochaeta gigantospora) that were potentially active against the Gram-positive and Gram-negative bacteria as well as the fungal pathogen, Candida albicans. The most prominent antagonistic activity against fungal pathogens was shown by P8-S4 (Tritirachium oryzae), P5-N31a (Truncatella spadicea), and P5-N20 (Fusarium larvarum). Our findings indicate that Pinus wallichiana harbors a rich endophytic fungal community with potential antimicrobial activities. Further studies are needed to understand the ecology and evolutionary context of the associations between the Himalayan pine and its endophytes.

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References

  1. Altschul SF, Madden TL, Schaffer AA, Zhang JH, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  2. Aly AH, Debbab A, Proksch P (2011) Fungal endophytes: unique plant inhabitants with great promises. Appl Microbiol Biotechnol 90:1829–1845

    Article  CAS  PubMed  Google Scholar 

  3. Anonymous (2008) A digest of forest statistics. Statistics Division, Department of Forests, Jammu and Kashmir Government, Srinagar, p 55

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

    Article  Google Scholar 

  5. 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 

  6. Arnold AE, Lutzoni F (2007) Diversity and host range of foliar fungal endophytes: are tropical leaves biodiversity hotspots? Ecology 88:541–549

    Article  PubMed  Google Scholar 

  7. Aslam M, Reshi Z, Siddiqi TO (2011) Genetic divergence in half-sib progenies of Pinus wallichiana A.B.. Jackson plus trees in the Kashmir Himalaya, India. Trop Ecol 52:201–208

    Google Scholar 

  8. Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (1994) Current protocols in molecular biology. Wiley, New York

    Google Scholar 

  9. Bacon CW, White JF (2000) Microbial endophytes. Dekker, New York, pp 341–388

    Google Scholar 

  10. Barnett HL, Hunter BB (1998) Illustrated genera of imperfect fungi, 4th edn. APS, St. Paul

    Google Scholar 

  11. Botella L, Diez JJ (2011) Phylogenic diversity of fungal endophytes in Spanish stands of Pinus halepensis. Fungal Divers 47:9–18

    Article  Google Scholar 

  12. Chang KK, Ju-Kyeong E, Ahn-Heum E (2013) Diversity and seasonal variation of endophytic fungi isolated from three conifers in Mt. Taehwa, Korea. Mycobiology 41:82–85

    Article  Google Scholar 

  13. Clardy J, Walsh C (2004) Lessons from natural molecules. Nature 432:829–837

    Article  CAS  PubMed  Google Scholar 

  14. Coppen JJW, Robinson JM, Kaushal AN (1988) Composition of xylem resin from Pinus wallichiana and P. roxburghii. Phytochemistry 27:2873–2875

    Article  CAS  Google Scholar 

  15. Dar AR, Dar GH (2006) The wealth of Kashmir Himalaya—gymnosperms. Asian J Plant Sci 5:251–259

    Article  Google Scholar 

  16. Dar MY, Shah WA, Mubashir S, Rather MA (2012) Chromatographic analysis, anti-proliferative and radical scavenging activity of Pinus wallichiana essential oil growing in high altitude areas of Kashmir, India. Phytomedicine 19:1228–1233

    Article  Google Scholar 

  17. Doty SL, Oakley B, Xin G, Kang JW, Singleton G, Khan Z, Vajzovic A, Staley JT (2009) Diazotrophic endophytes of native black cottonwood and willow. Symbiosis 47:23–33

    Article  CAS  Google Scholar 

  18. Ek-Ramos MJ, Zhou W, Valencia CU, Antwl JB, Kalns LL, Morgan GD, Kerns DL, Sword GA (2013) Spatial and temporal variation in fungal endophyte communities isolated from cultivated cotton (Gossypium hirsutum). PLoS ONE 8:e66049

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  19. Ellof JN (1998) A sensitive and quick microplate method to determine the minimal inhibitory concentration of plant extracts for bacteria. Planta Med 64:711–713

    Article  Google Scholar 

  20. Ezra D, Hess WH, Strobel GA (2004) New endophytic isolates of M. albus, a volatile antibiotic-producing fungus. Microbiology 150:4023–4031

    Article  CAS  PubMed  Google Scholar 

  21. Felsenstein J (1985) Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39:783–791

    Article  Google Scholar 

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

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  23. Ganley RJ, Newcombe G (2006) Fungal endophytes in seeds and needles of Pinus monticola. Mycol Res 110:318–327

    Article  PubMed  Google Scholar 

  24. Guo LD (2002) Pestalotiopsis besseyi, a new record of endophytic fungi from Pine in China. Mycosystema 21:455–456

    Google Scholar 

  25. Heywood VH (ed) (1995) Global biodiversity assessment. Cambridge University Press, Cambridge

    Google Scholar 

  26. Hoffman MT, Arnold AE (2008) Geographic locality and host identity shape fungal endophyte communities in cupressaceous trees. Mycol Res 112(Pt3):331–344

    Article  CAS  PubMed  Google Scholar 

  27. Iqbal J, Nelson JA, McCulley RL (2013) Fungal endophyte presence and genotype affect plant diversity and soil-to-atmosphere trace gas fluxes. Plant Soil 364:15–27

    Article  CAS  Google Scholar 

  28. Keller NP, Turner G, Bennett JW (2005) Fungal secondary metabolism—from biochemistry to genomics. Nat Rev Microbiol 3:937–947

    Article  CAS  PubMed  Google Scholar 

  29. Khosla C (1997) Harnessing the biosynthetic potential of modular polyketide synthases. Chem Rev 97:2577–2590

    Article  CAS  PubMed  Google Scholar 

  30. Kiffer E, Morelet M (2000) Deuteromycetes. Mitosporic fungi. Classification and generic keys. Science, Enfield

    Google Scholar 

  31. Korsten L, Jager ESD, Villiers EED, Lourens A, Kotze JM, Wehner FC (1995) Evaluation of bacterial epiphytes isolated from avocado leaf and fruit surfaces for biocontrol of avocado postharvest diseases. Plant Dis 79:1149–1156

    Article  Google Scholar 

  32. Kumar M, Qadri M, Sharma PR, Kumar A, Andotra SS, Kaur T, Kapoor K, Gupta VK, Kant R, Hamid A, Johri S, Taneja SC, Vishwakarma RA, Riyaz-Ul-Hassan S, Shah BA (2013) Tubulin inhibitors from an endophytic fungus isolated from Cedrus deodara. J Nat Prod 76:194–199

    Article  CAS  PubMed  Google Scholar 

  33. MacArthur R (1965) Patterns of species diversity. Biol Rev 40:510–533

    Article  Google Scholar 

  34. Minter DW, Millar CS (1980) Ecology and biology of three Lophodermium species on secondary needles of Pinus sylvestris. Eur J For Pathol 10:169–181

    Article  Google Scholar 

  35. Mousa WK, Raizada MN (2013) The diversity of anti-microbial secondary metabolites produced by fungal endophytes: an interdisciplinary perspective. Front Microbiol 4:65

    PubMed Central  PubMed  Google Scholar 

  36. Nagabhyru P, Dinkins RD, Wood CL, Bacon CW, Schardl CL (2013) Tall fescue endophyte effects on tolerance to water-deficit stress. BMC Plant Biol 13:127

    Article  PubMed Central  PubMed  Google Scholar 

  37. Osono T, Mori A (2004) Distribution of phyllosphere fungi within the canopy of giant dogwood. Mycoscience 45:161–168

    Article  Google Scholar 

  38. Porras-Alfaro A, Bayman P (2011) Hidden fungi, emergent properties: endophytes and microbiomes. Annu Rev Phytopathol 49:291–315

    Article  CAS  PubMed  Google Scholar 

  39. Qadri M, Johri S, Shah BA, Khajuria A, Sidiq T, Lattoo SK, Abdin MZ, Riyaz-Ul-Hassan S (2013) Identification and bioactive potential of endophytic fungi isolated from selected plants of the Western Himalayas. Springer Plus 2:8

    Article  PubMed Central  PubMed  Google Scholar 

  40. Reinhold-Hurek B, Hurek T (2011) Living in side plants: bacterial endophytes. Curr Opin Plant Biol 14:435–443

    Article  PubMed  Google Scholar 

  41. Rodriguez R, Redman R (2008) More than 400 million years of evolution and some plants still can’t make it on their own: plant stress tolerance via fungal symbiosis. J Exp Bot 59:1109–1114

    Article  CAS  PubMed  Google Scholar 

  42. Rodriguez RJ, Henson J, Van Volkenburgh E, Hoy M, Wright L, Beckwith F, Kim Y, Redman RS (2008) Stress tolerance in plants via habitat-adapted symbiosis. ISME J 2:404–416

    Article  PubMed  Google Scholar 

  43. Rudgers JA, Miller TE, Ziegler SM, Craven KD (2012) There are many ways to be a mutualist: endophyte fungus reduces plant survival but increases population growth. Ecology 93:565–574

    Article  PubMed  Google Scholar 

  44. Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425

    CAS  PubMed  Google Scholar 

  45. Schulz B, Guske S, Dammann U, Boyle C (1998) Endophyte host interactions II. Defining symbiosis of the endophyte host interaction. Symbiosis 25:213–227

    Google Scholar 

  46. Shannon CE, Weaver W (1963) The mathematical theory of communication. University of Illinois Press, Urbana

    Google Scholar 

  47. Sieber SA, Marahiel MA (2005) Molecular mechanisms underlying nonribosomal peptide synthesis: approaches to new antibiotics. Chem Rev 105:715–738

    Article  CAS  PubMed  Google Scholar 

  48. Simpson GG (1964) Species density of North American recent mammals. Syst Zool 13:57–73

    Article  Google Scholar 

  49. Sinclair WS, Lyon HH, Johnson WT (1987) Diseases of trees and shrubs, 4th edn. Cornell University Press, Ithaca, pp 32–33

    Google Scholar 

  50. Singh LP, Gill SG, Tuteja N (2011) Unraveling the role of fungal symbionts in plant abiotic stress tolerance. Plant Signal Behav 6:175–191

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  51. Staley JT, Castenholz RW, Colwell RR, Holt JG, Kane MD, Pace NR, Saylers AA, Tiedje JM (1997) The microbial world: foundation of the biosphere. American Academy of Microbiology, Washington, DC, p 32

    Google Scholar 

  52. Stone JK, Bacon CW, White JF Jr (2000) An overview of endophytic microbes: endophytism defined. In: Bacon CW, White JF Jr (eds) Microbial endophytes. Marcel Dekker, New York, pp 3–29

    Google Scholar 

  53. Strobel GA, Daisy B (2003) Bioprospecting for microbial endophytes and their natural products. Microbiol Mol Biol Rev 67:491–502

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  54. Tamura K, Nei M, Kumar S (2004) Prospects for inferring very large phylogenies by using the neighbor-joining method. Proc Natl Acad Sci 101:11030–11035

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  55. Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol Bio Evol 24:1596–1599

    Article  CAS  Google Scholar 

  56. Troup RS (1921) The silviculture of Indian trees. Vol.III. Clarendon, Oxford

    Google Scholar 

  57. Vandenkoornhuyse P, Baldauf SL, Leyval C, Straczek J, Young JP (2002) Extensive fungal diversity in plant roots. Science 295:2051

    Article  PubMed  Google Scholar 

  58. 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 315–322

    Google Scholar 

  59. Yuan Z, Zhang C, Lin F, Kubicek CP (2010) Identity, diversity, and molecular phylogeny of the endophytic mycobiota in the roots of rare wild rice (Oryza granulate) from a nature reserve in Yunnan, China. Appl Environ Microbiol 76:1642–1652

    Article  PubMed Central  CAS  PubMed  Google Scholar 

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Acknowledgments

The first author is grateful to the Indian Council of Medical Research, New Delhi for her Senior Research Fellowship. We acknowledge the financial support obtained from CSIR 12th FYP project, “PMSI” (BSC0117) of the Council of Scientific and Industrial Research (CSIR), New Delhi, India. The authors are also thankful to Mr. Fairoz I. Nazki, a resident of Lolab valley, for facilitating the collection of plant samples, and Mr. Jasbir Singh and Rajinder Kumar for preserving the cultures. This article bears the institutional manuscript number IIIM/1626/2013.

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Authors and Affiliations

  1. Microbial Biotechnology Division, CSIR—Indian Institute of Integrative Medicine, Canal Road, Jammu, 180001, India

    Masroor Qadri, Roopali Rajput, Ram A. Vishwakarma & Syed Riyaz-Ul-Hassan

  2. Centre for Transgenic Plant Development, Department of Biotechnology, Jamia Hamdard, New Delhi, 110062, India

    Malik Z. Abdin

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  1. Masroor Qadri
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Correspondence to Syed Riyaz-Ul-Hassan.

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Qadri, M., Rajput, R., Abdin, M.Z. et al. Diversity, Molecular Phylogeny, and Bioactive Potential of Fungal Endophytes Associated with the Himalayan Blue Pine (Pinus wallichiana). Microb Ecol 67, 877–887 (2014). https://doi.org/10.1007/s00248-014-0379-4

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