شناسایی ریخت شناختی و مولکولی قارچ های اندوفیت سرشاخه و شاخه‌ی درختان بلوط در جنگل های ارسباران

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانشجوی سابق کارشناسی ارشد، رشته بیماری شناسی گیاهی، گروه گیاه پزشکی، دانشکده کشاورزی، دانشگاه تبریز

2 استاد رشته قارچ شناسی و بیماری شناسی گیاهی، گروه گیاه پزشکی، دانشکده کشاورزی، دانشگاه تبریز

3 دانشجوی دوره دکتری، رشته قارچ شناسی و بیماری شناسی گیاهی، گروه گیاه پزشکی، دانشکده کشاورزی، دانشگاه تبریز

چکیده

چکیده
طی دو دهه­ی اخیر، علاقه­ی فزاینده­ای به مطالعه ی اندوفیت­ها، منشاء و تنوع زیستی آنها، اثرات متقابلشان و گیاهان میزبان، نقش آنها در اکولوژی و همچنین خصوصیات شیمیایی و فعالیت­های زیستی متابولیت­های ثانویه ی تولید شده توسط آنها وجود دارد. در این تحقیق قارچ­های اندوفیت سرشاخه و شاخه­ی بلوط درجنگل­های بلوط مناطق حاتم بیگ و کلیبر استان آذربایجانشرقی جداسازی شد و بر اساس ویژگی­های ریخت شناختی و داده­های مولکولی شناسایی گردید. بدین منظور در طی ماه­های تیر و شهریور 1393، از جنگل­های حاتم­بیگ مشگین­شهر و کلیبر نمونه­برداری صورت گرفت. هویت قارچ‌های جداسازی شده، پس از خالص­سازی به روش تک­اسپور و نوک­ریسه، با بررسی­های ریخت­شناختیوداده­های توالی ناحیه ITS-rDNA وTUB تعیین گردید. در این بررسی 25 گونه ی قارچی شامل Alternaria alternata species complex، Arthrinium arundinis، Aspergillus flavus،Biscogniauxia mediterranea، Chaetomium globosum، Cladosporium cladosporioides، Clonostachys rosea، Curvularia spicifera، Daldinia loculata، D. palmensis، D. vernicosa، Discula quercina، Epicoccum nigrum،Fusarium oxysporum ، F. proliferatum، F. solani،Nigrospora oryzae ، Ochrocladosporium elatum، Paecilomyces formosus، Penicillium commune، P. spinulosum، Pyronema domesticum،Sordaria fimicola.، S.sibutii و Trichothecium roseum از بافت­های پوست و چوب سرشاخه و شاخه­ی بلوط در جنگل­های این منطقه شناسایی شدند. همچنین گونه­های قارچی شناسایی شده در این تحقیق برای نخستین بار در دنیا از روی بلوط سیاه (Quercus macranthera) گزارش می شوند و بلوط سیاه به عنوان میزبانی جدید برای این گونه­ها معرفی می­گردد. حضور این قارچ­های اندوفیت در بافت­های به ظاهر سالم درختان بلوط می­تواند نشان­گر این امر باشد که برخی از این گونه­های قارچی ممکن است بیمارگر­های احتمالی نهفته گیاهی باشند.
 

 

کلیدواژه‌ها


عنوان مقاله [English]

Morphological and Molecular Characterization of Endophytic Fungi from Oak Trees in Arasbaran Forests

نویسندگان [English]

  • Saeid Ghasemi Esfahlan 1
  • Mehdi Arzanlou 2
  • Asadollah Babayahri 2
  • Mohsen Torbati 3
  • Abolfazl Narmani 3
1 1Formre MSc Student of Plant Pathology, Department of Plant Protection, Faculty of Agriculture, University of Tabriz, Tabriz, Iran.
2 Professor of Plant Pathology and Mycology Respectively, Department of Plant Protection, Faculty of Agriculture, University of Tabriz, Tabriz, Iran.
3 PhD Student of Plant Pathology, Department of Plant Protection, Faculty of Agriculture, University of Tabriz, Tabriz, Iran
چکیده [English]

Abstract
Over the past two decades, there has been an increasing interest in the study of endophytes, their origin, biodiversity, the interactions between endophytes and host plants, the role of endophytes in ecology, as well as the chemical properties and biological activity of secondary metabolites produced by them. In this research endophytic fungi from twigs and branches of oak in Hatam-baig and Kalibar regions were identified based on molecular and morphological characteristics. Towards this aim, samples were collected from twigs and branches of oak trees in above-mentioned regions during June and September 2014. Pure cultures were established using a single spore or hyphal tip techniques. The identity of fungal isolates was determined based on morphological characteristics and sequence data from ITS-rDNA and Tub region. In this study 25 fungal species including Alternaria alternata species complex, Arthrinium arundinis, Aspergillus flavus, Bipolaris spicifera, Biscogniauxia mediterranea, Chaetomium globosum, Clonostachys rosea, Cladosporium cladosporioides, Daldinia vernicosa, Daldinia palmensis, Daldinia loculata, Discula quercina, Epicoccum nigrum, Fusarium oxysporum, Fusarium proliferatum, Fusarium solani, Nigrospora oryzae, Ochrocladosporium elatum, Paecilomyces formosus, Penicillium commune, Penicillium spinulosum, Pyronema domesticum, Sordaria fimicola, Sordaria sibutii and Trichothecium roseum were identified as endophytic fungi inhabiting twigs and branches of oak trees in Hatam-baig and Kalibar region. The majority of the species that were identified in this study, are reported for the first time from Quercus macranthera. The assemblage of endophytic fungi in healthy tissues of oak trees might indicate that some of the fungi as possible latent pathogens of oak.
 

کلیدواژه‌ها [English]

  • Keywords: Endophyte
  • Hatam-baig and Kalibar
  • Black oak
  • ITS-rDNA
  • TUB
  • Arasbaran
Amal HA, Debbab A and Proksch P, 2011. Fungal endophytes: unique plant inhabitants with great promises. Applied Microbiology and Biotechnology 90: 1820-1845.
Arnold AE, Mejia LC, Kyllo D, Rojas EI, Maynard Z, Robbins N and Herre EA, 2003. Fungal endophytes limit pathogen damage in a tropical tree. Proceedings of the National Academy of Sciences USA 100: 15649-15654.
Blumenstein K, 2010. Characterization of endophytic fungi in the genus Ulmus: putative agents for the biocontrol of Dutch elm disease (DED). Diploma thesis, University of Kassel, Germany.
Cai L, Jeewon R and Hyde KD. 2006. Phylogenetic investigations of Sordariaceae based on multiple gene sequences and morphology. Mycological Research. 110(2): 137-150.
Collado J, Platas G and Pelaez F, 1996. Fungal endophytes in leaves, twigs and bark of Quercus ilex from central Spain. Nova Hedwigia 63: 347-360.
Crous PW and Groenewald jz, 2013. A phylogenetic re-evalution of Arthrinium. IMA Fungus. 4 (1): 133-145.
Ershad D, 2009. Fungi of Iran. 3rd edition, Iranian Research Institution of Plant Protection, 531 p.
Fisher PJ, Petrini O, Petrini LE and Sutton BC, 1994. Fungal endophytes from the leaves and twigs of Quercus ilex L. from England, Majorca and Switzerland. New Phytologist 127: 133-137.
Gennaro M, Gonthier P and Nicolotti G, 2003. Fungal endophytic communities in healthy and declining Quercus robur L. and Q. cerris L. trees in northern Italy. Journal of Phytopathology 151: 529-534.
Ghasemi Esfahlan S, Arzanlou M and Babai Ahari A, 2017. Identification of endophytic Trichoderma species from oak trees in Arasbaran forests using morphological and molecular characteristics. Applied Research in Plant Protection. 6 (3): 53-66.
Ghasemi Esfahlan S, Arzanlou M, Khoadei S, Tavakoli M and Babai Ahari A, 2016. Molecular diagnostics of Biscogniauxia mediterranea, the causal agent of charcoal rot disease on oak using species-specific primers. Applied Research in Plant Protection. 5 (2): 175-186.
Hajizadeh A, Amini J and Abdollahzadeh J, 2015. New records of endophytic fungi isolated from oak trees in Kurdistan province (Iran). Rostaniha. 16 (1):109-122.
Halmschlager E, Butin H and Donaubauer E, 1993. Endophytic fungi in leaves and twigs of Quercus petraea. European Journal of Forest Pathology 23: 51-63.
 Helander M, Ahlholm J, Sieber TN, Hinneri S and Saikkonen K, 2007. Fragmented environment affects birch leaf endophytes. New Phytologist 175: 547-553.
Huhndorf SM, Miller AN and Fernandez FA, 2004. Molecular systematics of the Sordariales: the order and the family Lasiosphaeriaceae redefined. Mycologia. 96(2): 368-387.
James TY, Kauff F, Schoch CL, Matheny PB, Hofstetter V, Cox CJ, Celio G, Gueidan C, Fraker E, Miadlikowska J, Lumbsch HT, Rauhut A, Reeb V, Arnold AE, Amtoft A, Stajich JE, Hosaka K, Sung GH, Johnson D, O’Rourke B, Crockett M, Binder M, Curtis JM, Slot JC, Wang Z, Wilson AW, Schussler A, Longcore JE, O’Donnell K, Mozley-Standridge S, Porter D, Letcher PM, Powell MJ, Taylor JW, White MM, Griffith GW, Davies DR, Humber RA, Morton JB, Sugiyama J, Rossman AY, Rogers JD, Pfister DH, Hewitt D, Hansen K, Hambleton S, Shoemaker RA, Kohlmeyer J, Volkmann-Kohlmeyer B, Spotts RA, Serdani M, Crous PW, Hughes KW, Matsuura K, Langer E, Langer G, Untereiner WA, Lucking R, Budel B, Geiser DM, Aptroot A, Diederich P, Schmitt I, Schultz M, Yahr R, Hibbett DS, Lutzoni F, McLaughlin DJ, Spatafora JW andVilgalys R, 2006. Reconstructing the early evolution of fungi using a six-gene phylogeny. Nature 443: 818-822.
Ju YM and Rogers JD, 1996. A revision of the genus Hypoxylon.Mycologia Memoir.20: 1-365
Lorenzi E, Lorando E and Picco AM, 2006. Microfunghi endofitici ed epifitici di Picea abies (L.) Karst. in ambiente naturale ed antropizzato in Lombardia. Forest Pathology. 3: 426-436.
Masoudinejad M R and Rezazade M, 2003. Comparison of four methods extraction from the fruits of oak species in Iran. Hakim 1: 81-91.
Moller E M, BahnwegG, SandermanH and Geiger HH, 1992. A simple and efficient protocol for isolation of high molecular weight DNA from filamentous fungi, fruit bodies and infected plant tissues. Nucleic Acids Research 20: 6115-6116.
Nylander JAA, 2004. MrModeltest v. 2.0. Program distributed by the author. Evolutionary Biology Centre, Uppsala University, Uppsala, Sweden.
Ragazzi A, Moricca S, Capretti P, Dellavalle I and Turco E, 2003. Differences in composition of endophytic mycobiota in twigs and leaves of healthy and declining Quercus species in Italy. Forest Pathology 33: 31-38.
Ragazzi A, Vagniluca S and Moricca S, 1995, European expansion of oak decline: involved microorganisms and methodological approaches. Phytopathologia Mediterranea.34: 207 226.
Ronquist F and Huelsenbeck JP, 2003. MrBayes 3: Bayesian phylogenetic inference under mixed models. Bioinformatics 19: 1572-1574.
Saikkonen K, Faeth SH, Helander M and Sullivan TJ, 1998. Fungal endophytes: a continuum of interactions with host plants. Annual Review of Ecology and Systematics 29: 319-343.
Schulz B and Boyle C, 2005. Review: The endophytic continuum. Mycological Research 109: 661-686.
Sharma R, Kulkarni G, Sonawane MS and Shouche YS, 2014. Anew endophytic species of Arthrinium (Apiosporaceae) from Jatropha podagrica. Mycoscience. 55 (2): 118-123.
Soltis PS and Soltis DE. 2000. The role of genetic and genomic attributes in the success of polyploids. Proceedings of the National Academy of Sciences of the United States of America. 97: 7051-7057.
Tamura K, Stecher G, Peterson D, Filipski A and Kumar S, 2013. MEGA 6: molecular evolutionary genetics analysis version 6.0. Molecular Biology and Evolution 30: 2725-2729.
Tan RX and Zou WX, 2001. Endophytes: a rich source of functional metabolites. Natural Product Reports 18: 448-459.
Thomas N, 2007. Endophytic fungi in forest trees: are they mutualists? Fungal biology reviews 75- 89.
Zabalgogeazcoa I, 2008. Fungal endophytes and their interaction with plant pathogens. Spanish Journal of Agricultural Research 6: 138-146.