شناسایی گونه های تریکودرمای اندوفیت درختان بلوط در جنگل های ارسباران با استفاده از معیارهای ریخت شناختی و مولکولی

نوع مقاله : مقاله پژوهشی

نویسندگان

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

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

چکیده

چکیده
گونه­های جنس Trichoderma از پراکنش جهانی بر­خوردار بوده و مایکوبیوتای غالب خاک­های مناطق مختلف می­باشند. گونه­های تریکودرما علیرغم این­که از پتانسیل ساپروفیتی قوی برخوردار می­باشند، دارای فاز اپیفیتکی قابل توجهی بوده و از پتانسیل نفوذ به بافت­های داخلی میزبان گیاهی بدون ایجاد خسارت و صدمه به میزبان برخوردار هستند. در سال­های اخیر، توجه ویژه­ای به شناسایی و برهمکنش گونه­های تریکودرمای اندوفیت با میزبان­های گیاهی معطوف گردیده است، با این وجود، اطلاعات چندانی در مورد تنوع زیستی گونه­های تریکودرمای اندوفیت در ایران وجود ندارد. تحقیق حاضر با هدف شناسایی گونه­های تریکودرمای اندوفیت درختان بلوط در جنگل­های ارسباران انجام گردید. برای این منظور در تابستان سال  1393 از قسمت­های تنه و سرشاخه­ی درختان بلوط سالم و درختان دارای علایم زوال در مناطق حاتم بیگ )مشگین شهر) و کلیبر نمونه­برداری شد. در مجموع، 23 جدایه­ی­ تریکودرما از مناطق نمونه­برداری شده جداسازی و خالص­سازی گردید. هویت جدایه­ها بر اساس ویژگی­های ریخت­شناختی و داده­های توالی ناحیه ITS-rDNA، Trichoderma atroviride، T. citrinoviride، T. harzianum، T. longibrachiatum و T. polysporum تعیین گردید. در تبار­نما­ی ترسیم شده براساس توالی ناحیه ITS-rDNA، جدایه­های مربوط به هر یک از گونه­های شناسایی شده به همراه توالی نمونه­ی تیپ هریک از گونه­ها، با اعتبار سنجی بالا گروه­بندی شدند.  کلیه­ی گونه­ها برای اولین بار از جنگل­های ارسباران گزارش می­شوند و بلوط سیاه (Quercus macranthera) به عنوان میزبان جدید برای تمامی گونه­های شناسایی شده در این تحقیق معرفی می شود. گزارش این گونه­ها ­از روی بلوط سیاه برای دنیا جدید می­باشد. با شناسایی این گونه­ها امکان ارزیابی کارایی این گونه­ها در کنترل زیستی بیمارگرهای گیاهی و تقویت رشد گیاهان فراهم خواهد شد.

 

کلیدواژه‌ها


عنوان مقاله [English]

Identification of Endophytic Trichoderma Species From Oak Trees in Arasbaran Forests Using Morphological and Molecular Characteristics

نویسندگان [English]

  • Saeeid Ghasemi Esfahlan 1
  • Mehdi Arzanluou 2
  • Asaloulah Babaei Ahari 2
1 Former 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.
چکیده [English]

Abstract
Trichodermaspecies have a global distribution, representing dominant mycobiota of soils in different regions. Even though Trichodermaspecies exhibit strong saprophytic potential, they possess significant epiphytic phase and can penetrate internal tissues of plants without causing any harm to host plant.  In recent years, substantial attention has been paid on the identification and interaction of endophytic Trichoderma species with plant hosts. However, there is a huge paucity of knowledge on biodiversity of endophytic Trichoderma species in Iran. The present study was aimed to characterize endophytic Trichoderma species from oak trees in Arasbaran forests. For this purpose, during August-September 2014, samples from twig and trunk of oak trees were collected in the Hatam-baig (Meshgin-Shahr) and the Kaleibar regions. A total number of 23 Trichoderma isolates were recovered from sampled areas. Trichoderma isolates were characterized based on morphological data and sequence data of ITS-rDNA region as Trichoderma atroviride,T. citrinoviride,T.harzianum, T. longibrachiatumand T. polysporum. In a phylogeny inferred based on sequence data of ITS-rDNA region, Trichoderma spp were clustered with the representative sequence of the type strain for each of the species with high bootstrap support. All five species are newly recorded from Arasbaran forests and black oak (Quercus macranthera) is reported as new host for these species. With the identification of these species, it will be possible to evaluate their efficacy in control of plant pathogens and promotion of plant growth.
 

کلیدواژه‌ها [English]

  • Keywords: Endophyte
  • Black oak
  • Trichoderma
  • ITS-rDNA
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.
Bailey BA and Lumsdon RD, 1998. Direct effects of Trichoderma and Gliocladium, In: Harman GE,  Kubicek CP (eds.), Trichoderma and Gliocladium: enzymes, biological control and commercial applications. Taylor & Francis, Bristol, PA, pp. 185-204.
Bailey BA and Melnick RL, 2013. The endophytic Trichoderma. In: Mukherjee PK, Horwitz BA, Singh US, Mukherjee M, Schmoll M (eds.), Trichoderma: biology and applications, 1st edn. CAB International, London, pp. 152-172.
Bills GF and Polishook JD, 1991. Microfungi from Carpinus caroliniana. Canadian Journal of Botany 69: 1477-1482.
Bisset J, 1984. A revision of the genus Trichoderma. I. Section Longibrachiatum sect. nov. Canadian Journal of Botany 62: 924-931.
Bisset J, 1991. A revision of the genus Trichoderma. II. Infrageneric classification. Canadian Journal of Botany 69: 2357–2372.
Bissett J, 1992. Trichoderma atroviride. Canadian Journal of Botany 70: 639-641.
Bissett J, Gams W, Jaklitsch W, Samuels GJ, 2015. Accepted Trichoderma names in the year 2015. IMA Fungus 6: 263-295.
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.
Bultman TL and Murphy JC, 2000. Do fungal endophytes mediate wound-induced resistance? In: Bacon CW, White JF Jr, (eds.), Microbial endophytes. Marcel Dekker, pp: 421-453.
Chaverri P, Samuels GJ, 2003. Hypocrea/Trichoderma (Ascomycota, Hypocreales, Hypocreaceae): species with green ascospores. Studies in Mycology 48: 1–116.
Dehghanpour FS, Sharifnabi B and Mirlohi AF, 2006. Application of 5.8 S gene and ITS, PCR-RFLP pattern in taxonomy of Neotyphodium endophytic fungi. Rostaniha 7(1): 1-15.
Ershad D, 2009. Fungi of Iran. 3rd edition, Iranian Research Institution of Plant Protection, 531 p.
Farr DF, Rossman AY. 2017. Fungal Databases, U.S. National Fungus Collections, ARS, USDA. Retrieved January, 12, 2017, from https://nt.ars-grin.gov/fungaldatabases/
Gams W and Bissett J, 1998. Morphology and identification of Trichoderma In: Kubicek Trichoderma and Gliocladium 1: 3-34.
Hanada RE, De Souza JT, Pomella AWV, Hebbar KP, Pereira JO, Ismaiel A and Samuels GJ , 2008. Trichoderma martiale sp. nov., a new endophyte from sapwood of Theobroma cacao and a potential agent of biological control. Mycological Research 112: 1335-1343.
Helander M, Ahlholm J, Sieber TN, Hinneri S and Saikkonen K, 2007. Fragmented environment affects birch leaf endophytes. New Phytologist 175: 547-553.
Jaklitsch WM, Samuels GJ and Dodd SL, 2006. Hypocrea rufa / Trichoderma viride: a reassessment and description of five closely related species with and without warted conidia. Studies in Mycology 56: 135-177.
Klein D and Everleigh DE, 1998. Ecology of Trichoderma, In: Kubicek CP, Harman GE (eds.). Trichoderma and Gliocladium. Taylor & Francis, London, UK.  pp: 57-74.
Kubicek CP and  Penttila ME, 1998. Regulation of production of plant polysaccharide degrading enzymes by Trichoderma In: Kubicek CP, Harman GE (eds.). Trichoderma and Gliocladium: enzymes, biological control and commercial applications. Tailor & Frrancis Ltd. London UK.  pp. 49-71.
Lucia M, Annalisa C, Antonio F, Benedetto TL, Salvatore C, Tonina R and Daniela P, 2009. Occurrence and characterization of peptaibols from Trichoderma citrinoviride, an endophytic fungus of cork oak, using electrospray ionization quadrupole time- of- flight mass spectrometry. Microbiology 155: 3371-3381.
Malinowski DP and Belesky DP, 2000. Adaptation of endophyte infected cool-season grasses to environmental stresses. Crop Science 40: 923-940.
Mirlohi AF, Sabzalian MR and Khayam nekoie M, 2004. Endophytic fungi, characteristics and their potential for genetic manipulation. Iranian Journal of Biotechnology 2: 75-83.
Moller EM, Bahnweg G, Sanderman H 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.
Parsaeian M, Mirlohi AF, Rezaie AM and Khayam nekoie M, 2007. The effect of endophytic fungi on physiological characteristics and cold tolerance of two species of meadow fescue and tall fescue. Journal of Science and Technology of Agriculture and Natural Resources 10: 197-212.
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.
Rifai MA, 1969. A revision of the genus Trichoderma. Mycologia 116: 1-56.
Roghanian M, Amini J, Abdollahzadeh J and Zafari D, 2013. A record of endophytic Trichoderma species in Kurdistan. Plant Protection Journal 5: 115-124.
Ronquist F and Huelsenbeck JP, 2003. MrBayes 3: Bayesian phylogenetic inference under mixed models. Bioinformatics 19: 1572-1574.
Rossman AY, 1996. Morphological and molecular perspectives on systematics of Hypocreales. Mycologia 88: 1-19.
Sabzalian MR, Hatami B and Mirlohi AF, 2004. Mealybug, Phenocccous solani, and barley aphid, Sipha maydis, response to endophyteinfected tall and, meadow fescues. Entomologia Experimentalis et Applicata 113: 205-209.
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.
Samuels GJ, 1996. Trichoderma: a review of biology and systematics of the genus. Mycological Research 100: 923-935.
Samuels GJ and Ismaiel A, 2009. Trichoderma evansii and T. lieckfeldtiae: two new T. hamatum-like species. Mycologia 101: 142-156.
Samuels GJ, Ismaiel A, Mulaw TB, Szakacs G, Druzhinina IS, Kubicek CP, Jaklitsch WM, 2012. The Longibrachiatum clade of Trichoderma: a revision with new species. Fungal Diversity 55: 77-108.
Schulz B and  Boyle C, 2005. The endophytic continuum. Mycological Research 109: 661-686.
Sivasithamparam K and Ghisalberti EL, 1998. Secondary metabolism in Trichoderma and Gliocladium, In: Kubicek CP, Harman GE (eds.). Trichoderma and Gliocladium: basic biology, taxonomy and genetics. Taylor and Francis Ltd. London, UK. pp: 131-191.
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.
White JFJ and Torres MS, 2010. Is plant endophyte-mediated defensive mutualism the result of oxidative stress protection?. Physiologia Plantarum 138: 440-446.
White TJ, Bruns T, Lee S and 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 Press, San Diego. pp: 315-322.
Zabalgogeazcoa I, 2008. Fungal endophytes and their interaction with plant pathogens. Spanish Journal of Agricultural Research 6: 138-146.