Türkiye’nin kuzeybatısındaki bir plantasyonda yerli ve egzotik çam türlerinin ibrelerinde görülen fungal endofitler

Abstract

İzmit Kerpe Araştırma Ormanı'nda 2016 yılında Pinus sylvestris, Pinus pinaster, Pinus nigra, Pinus taeda ve Pinus radiata ib-relerinde bulunan endofitik funguslar araştırılmıştır. Her çam türünden on ağacın tepe tacının alt kısmından sistematik olarak önceki yılın ibreleri örneklenmiştir. Her bir ibre yüzeysel olarak steril edilmiş ve 0,5 cm büyüklüğünde parçalara ayrılmış daha sonra Malt Ekstrakt Agar besi ortamına yerleştirilmiştir. Gelişen izolatlardan seçilen temsili 7 izolattan DNA ekstraksiyonu ya-pılarak, 5.8S rDNA, 18S ve 28S rDNA genleri arasında yer alan iki ITS bölgesini hedef alan ITS1 ve ITS4 primerleri kullanıla-rak amplifiye edilmiştir. Amplikonlar, evrensel fungus primerleri kullanılarak her iki yönde dizilenmiştir. Toplam 1000 ibreden (her çam türünden 200 ibre) yapılan izolasyonda 750 fungus izolatı elde edilmiştir. En fazla oranda Pestalotiopsis funerea izole edilmiş, bunu Acremonium sp., Cladiosporum sp. ve Cyclaneusma minus takip etmiştir.

Keywords

• Fungal topluluk
• Marmara Bölgesi
• ITS bölgesi

Fungal endophytes in the needles of native and exotic pine species in a planta-tion in Northwestern Türkiye

Abstract

The endophytic fungi present in needles of Pinus sylvestris, Pinus pinaster, Pinus nigra, Pinus taeda and Pinus radiata were investigated in Kerpe Research Forest, İzmit in 2016. Ten trees of each pine species were sampled systematically. Previous years green needles were sampled from the lower part of the canopy, from two equally spaced positions around the tree. Each needle was surface sterilized and cut into 0.5 cm sections then individually placed onto malt extract agar plates. DNA was ex-tracted from reprensentative seven isolates and amplified using primers ITS1 and ITS4 targeting the nuclear 5.8S rDNA gene and the two ITS regions flanked between 18S and 28S rDNA genes. Amplicons sequenced in both directions using the univer-sal fungal primers ITS1 and ITS4. Isolations from a total of 1000 needles (200 from each pine species) yielded 750 fungal isolates. Pestalotiopsis funerea (Desm.) Steyaert was isolated at the highest frequency followed by Acremonium sp., Cladio-sporum sp. and Cyclaneusma minus.

Keywords

• Fungal community
• Marmara Region
• ITS region

References

1. Agrios, G.N., 1988. Plant Pathology. Academic Press, New York, USA.
2. Arnold, A.E., Henk, D.A., Eells, R.L., Lutzoni, F., Vilgalys, R., 2007. Diversity and phylogenetic affinities of foliar fungal endophytes in loblolly pine inferred by culturing and envi-ronmental PCR. Mycologia, 99: 185-206.
3. Bills, G.F., 1996. Isolation and analysis of endophytic fungal communities from woody plants. See Reference, 124: 31–65.
4. Brown, K.B., Hyde, K.D., Guest, D.I., 1998. Preliminary studies on endophytic fungal communities of Musa acuminata spe-cies complex in Hong Kong and Australia. Fungal Diversity, 1: 27-51.
5. Botella, L., Dize, J.J., 2011. Phylogenic diversity of fungal endo-phytes in Spanish stands of Pinus halepensis. Fungal Diversity, 47:9-18.
6. Collado, J., Platas, G., Peláez, F., González, I., Peláez, F., 1999. Geographical and seasonal influences on the distribution of fungal endophytes in Quercus ilex L. New Phytologist, 144: 525–532.
7. Collado, J., Platas, G., Peláez, F., González, I., Peláez, F., 2000. Host specificity in fungal endophytic populations of Quercus ilex and Quercus faginea from Central Spain. Nova Hedwigia, 71: 42–430.
8. Danti, R., Sieber, T.N., Sanguineti, G., 2002. Endophytic mycobi-ota in bark of European beech (Fagus sylvatica) in the Apen-nines. Mycological Research, 106: 1343-1348.

9. El-Morsy, E.M., Dohlob, S.M., Hyde, K.D., 2006. Diversity of Alternaria alternata a common destructive pathogen of Eich-hornia crassipes in Egypt and its potential use in biological control. Fungal Diversity, 23: 139-158.
10. Ercan, M., 2002. İzmit- Kerpe Araştırma Ormanı. Çevre ve Or-man BakanlığıTeknik Bülten 213 (15): 114s.
11. Ganley, R.J., Brunsfeld, S.J., Newcombe, G., 2004. A community of unknown, endophytic fungi in western white pine. Pro-ceedings of the National Academy of Sciences, USA 101: 10107–10112.
12. Giordano, L., Gonthier, P., Varese, G.C., Miserere, L., 2009. Mycobiota inhabiting sapwood of healthy and declining Scots pine (Pinus sylvestris L.) trees in the Alps. Fungal Diversity, 69-83.
13. Gou, L.D., Huang, G.R., Wang, Y., 2008. Seasonal and tissue age influences on endophytic fungi of Pinus tabulaeformis (Pina-ceae) in the Dongling Mountains, Beijing. Journal of Integra-tive Plant Biology, 50(8): 997–1003.
14. Göre, M.E., Bucak, C., 2007. Geographical and seasonal influ-ences on the distribution of fungal endophytes in Laurus no-bilis. Forest Pathology, 37: 281-288.
15. Hata, K., Futai, K., Tsuda, M., 1998. Seasonal and needle age-dependent changes of the endophytic mycobiota in Pinus thunbergii and Pinus densiflora needles. Can J Bot, 76:245–250.
16. Jumpponen, A., Jones, K.L., 2009. Massively parallel 454 se-quencing indicates hyperdiverse fungal communitiesin tem-perate Quercus macrocarpa phyllosphere. New Phytologist, 184: 438-448.
17. Jurc, D., Jurc, M., Sieber, T.N., Bojovic, S., 2000. Endophytic Cenangium ferruginosum (Ascomycota) as a reservoir for an epidemic of Cenangium dieback in Austrian pine. Phyton-Annales Rei Botanicae, 40: 103–108.
18. Kirk, P.M., Cannon, P.F., David, J.C., Stalpers, J.A., 2001. Ains-worth & Bisby's Dictionary of the Fungi, 9th Edition. CABI Publishing.
19. Kowalski, T., 1982. Fungi infecting Pinus sylvestris needles of various ages. European Journal of Forest Pathology, 12: 182-190.
20. Kowalski, T., 1993. Fungi in living symptomless needles of Pinus sylvestris with respect to some observed disease processes. Journal of Phytopathology, 139: 129-145.
21. Kowalski, T., Zych, P., 2002. Endophytic fungi in needles of Pinus nigra growing under different site conditions. Polish Botanical Journal, 47 (2(: 251-257.
22. Lehtijärvi, A. T., Barklund P., 2000. Seasonal patterns of coloni-zation of Norway spruce needles by Lophodermium piceae. Forest Pathology, 30: 185-193.
23. Lehtijärvi, A., Oskay, F., Aday, A.G., Doğmuş Lehtijärvi, H.T., 2010. Isparta- Yenişarbademli ilçesi konifer ormanlarında ib-re ve sürgün hastalıklarına neden olan fungal etmenlerin tes-piti. III. Ulusal Karadeniz Ormancılık Kongresi, 20-22 Mayıs, Trabzon, s. 1420-1430.
24. Martín, P., Pajares, J.A., Pando, V., Diez, J.J., 2006. Fungi isolated from diseased nursery seedlings in Spain. New Forests, 31: 41-56.
25. Minter, D.W., Millar, C.S., 1980. Ecology and biology of three Lophodermium species on secondary needles of Pinus syl-vestris. European Journal of Forest Pathology, 10: 169- 181.
26. Müller, M.M., Hallaksela, A.M., 2000. Fungal diversity in Nor-way spruce: a case study. Mycological Research, 104(9):1139-1145.
27. Petrini, O., 1991. Fungal endophytes in tree leaves, In: Microbial Ecology of Leaves (eds. J.H. Andrews and S.S Hiraro) Springer New York, pp. 179-197.
28. Rodriguez, R.J., White, J.F., Arnold, A.E., Redman, R.S., 2009. Fungal endophytes: diversity and functional roles. New phy-tologist, 182(2): 314-330.
29. Rojas, E.I, Rehner, S.A., Samuels, G.J., Van Bael, S.A., Herre, E.A., 2010. Colletotrichum gloeosporioides s.l. associated with theobroma cacao and other plants in Panama: multilocus phylogenies distinguish host associated pathogens from asymptomatic endophytes. Mycologia, 102: 1318-1338.
30. Santamaría, O., Díez, J.J., 2005. Fungi in leaves, twigs and stem bark of Populus tremula from northern Spain. Forest Patholo-gy, 35: 95– 104.
31. Santamaría, O., Tejerina, L., Pajares, J.A., Diez, J.J., 2006. Effects of associated fungi Sclerophoma pythiophila and Cenangium ferruginosum on Gremmeniella abietina dieback in Spain. Forest Pathology, 36: 1-8.
32. Sieber, T., 2007. Endophytic fungi in forest trees: are they mutu-alists? Fungal Biology Reviews, 21:75-89.
33. Sinclair, W.A., Lyon, H., 1987. Diseases of trees and shrubs. Comstock Publishing Associates, London.
34. Suryanarayanan, T.S., Senthilarasu, G., Muruganandam, V., 2000. Endophytic fungi from Cuscuta reflexa and its host plants. Fungal Divers, 4:117–123.
35. Tokumasu, S., 1978. Leaf litter fungi of the forests of Pinus densiflora and four introduced pines at Sugadaira, central Ja-pan. Transactions of the Mycolological Society of Japan, 19: 383-390.
36. Wei, J.G., Xu, T., Guo, L.D., Liu, A.R., Zhang, Y., Pan, X.H., 2007. Endophytic Pestalotiopsis species associated with plants of Podocarpaceae, Theaceae and Taxaceae in southern China. Fungal Diversity, 24: 55-74.
37. White, T., Bruns, T., Lee, S., Taylor, J., 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylo-genetics. In: PCR Protocols, a Guide to Methods and Applica-tions (Ed: Innis, M.A., Gelfand, D.H., Sninsky, J.J., White, T.J.), Academic Press, New York, pp. 315-322.
38. Zamora, P., Martínez-Ruiz, C., Diez, J.J., 2008. Fungi in needles and twigs of pine plantations from northern Spain. Fungal Di-vers, 30:171–184.