Polyphasic identification of Cercospora cf. sigesbeckiae as causal agent of cercospora leaf spot of oilseed plants soybean, sesame and rapeseed in Golestan and Mazandaran provinces

Document Type : Research Article

Authors

1 Department of Botany, Iranian Research Institute of Plant Protection, Agricultural Research, Education and Extension Organization (AREEO), Tehran, Iran

2 Iranian Research Institute of Plant Protection, Agricultural Research, Education and Extension Organization (AREEO), Tehran, Iran

Abstract

گیاهان زراعی سویا، کلزا و کنجد از گیاهان صنعتی مهم در دنیا و ایران به شمار می‌روند. بیماری لکه‌برگی سرکوسپورایی یکی از تهدیدات جدی تولید این گیاهان به‌ویژه گیاه سویا در دنیا به حساب می‌آید. طی بازدید از مزارع گیاهان زراعی مختلف در استان‌های گلستان و مازندران، شیوع گسترده‌ای از علایم لکه‌برگی سرکوسپورایی روی گیاهان سویا، کلزا و کنجد مشاهده شد. به منظور شناسایی عوامل دخیل در این بیماری، از برگ‌های دارای علایم لکه-برگی، نمونه‌برداری به عمل آمد و کشت‌های تک اسپور به صورت مستقیم از لکه‌های برگی تهیه شدند. در مجموع تعداد 50 جدایه قارچی به دست آمد که از بین آنها هشت جدایه از گیاه سویا، چهار جدایه از کنجد و چهار جدایه از کلزا انتخاب و با استفاده از ترکیب خصوصیات ریخت‌شناختی و توالی شش ناحیه ژنی ITS، tef1، actA، cmdA ، his3 و gapdh شناسایی شدند. نتایج تجزیه و تحلیل فیلوژنتیکی توالی‌ها نشان داد که گونه Cercospora cf. sigesbeckiae به عنوان عامل لکه‌برگی سرکوسپورایی در این دو استان روی سه گیاه زراعی سویا، کنجد و کلزا فعالیت می‌نماید. بر اساس منابع موجود، تحقیق حاضر اولین گزارش از وقوع گونه‌ C. cf. sigesbeckiae از ایران و اولین گزارش این گونه روی کلزا و کنجد در دنیا می‌باشد.

Keywords


Ahmadi K., Ebadzadeh H., Abdeshah H., Kazemian A. and Rafee M. 2018. Statistics of agricultural products; crops. Ministry of Jihad-e-Agriculture,Tehran, Iran. (in Persian)
Albu S., Schneider R., Price P. and Doyle V. 2016. Cercospora cf. flagellaris and Cercospora cf. sigesbeckiae are associated with Cercospora leaf blight and purple seed stain on soybean in North America. Phytopathology106: 1376–85.
Bakhshi M. 2018. Important criteria for identification of the Cercospora species. Plant Pathology Science 7 (1): 1–14.
Bakhshi M. 2019. Epitypification of Cercospora rautensis, the causal agent of leaf spot disease on Securigera varia, and its first report from Iran. Fungal Systematics and Evolution 3: 157–163.
Bakhshi M. and Arzanlou M. 2017. Multigene phylogeny reveals a new species and novel records and hosts in the genus Ramularia from Iran. Mycological Progress16: 703–712.
Bakhshi M., Arzanlou M. and Babai-Ahari A. 2011. Uneven distribution of mating type alleles in Iranian populations of Cercospora beticola, the causal agent of Cercospora leaf spot disease of sugar beet. Phytopathologia Mediterranea 50: 101–109.
Bakhshi M., Arzanlou M. and Babai-Ahari A. 2012. Morphological and molecular characterization of Cercospora zebrina from black bindweed in Iran. Plant Pathology and Quarantine2: 125–130.
Bakhshi M., Arzanlou M., Babai-ahari A., Groenewald J.Z. and Crous P.W. 2018. Novel primers improve species delimitation in Cercospora. IMA Fungus 9(2): 299–332.
Bakhshi M., Arzanlou M., Babai-ahari A., Groenewald J.Z., Braun U. and Crous P.W. 2015. Application of the consolidated species concept to Cercospora spp. from Iran. Persoonia34: 65–86.
Bakhshi M., Zare R., Arzanlou M. and Narmani A. 2020. Occurrence of Cercospora leaf spot disease on Cucumis melo in Guilan province and characterization of the casual agent(s) using molecular and morphological approaches. Iranian Journal of Plant Pathology 55(4): in press. (in Persian)
Carbone I. and Kohn L.M. 1999. A method for designing primer sets for speciation studies in filamentous ascomycetes. Mycologia 91: 553–556.
Crous P.W. and Braun U. 2003. Mycosphaerella and its anamorphs: 1. Names published in Cercospora and Passalora. CBS Biodiversity Series 1: 1–571.
Crous P.W., Groenewald J.Z., Groenewald M., Caldwell P., Braun U. and Harrington T.C. 2006. Species of Cercospora associated with grey leaf spot of maize. Studies in Mycology 55: 189–197.
Crous P.W., Groenewald J.Z., Mansilla J.P., Hunter G.C. and Wingfield M.J. 2004. Phylogenetic reassessment of Mycosphaerella spp. and their anamorphs occurring on Eucalyptus. Studies in Mycology 50: 195–214.
de Hoog G.S. and Gerrits van den Ende A.H.G. 1998. Molecular diagnostics of clinical strains of filamentous Basidiomycetes. Mycoses 41: 183–189.
Duangsong U., Laosatit K., Somta P. and Srinives P. 2018. Genetics of resistance to Cercospora leaf spot disease caused by Cercospora canescens and Psuedocercospora cruenta in yardlong bean (Vigna unguiculata ssp. sesquipedalis) x grain cowpea (V. unguiculata ssp. unguiculata) populations. Journal of Genetics 97: 1451–1456.
Ershad D. 2009. Fungi of Iran. Ministry of Jihad-e-Agriculture. Agricultural Research, Education and Extension Organization, Tehran, Iran.
Ershad D., Asef M.R., Bakhshi M., Javadi A., Zangeneh S., Asgari B., Aliabadi F. and Mehrabi M. 2018. Genera of Fungi and Fungal Analogues of Iran. Ministry of Jihad-e-Agriculture. Agricultural Research, Education and Extension Organization, Tehran, Iran. (in Persian)
Groenewald J.Z., Nakashima C., Nishikawa J., Shin H.-D., Park J.H., Jama A.N., Groenewald M., Braun U. and Crous P.W. 2013. Species concepts in Cercospora: spotting the weeds among the roses. Studies in Mycology 75: 115–170.
Groenewald M., Groenewald J.Z., Braun U. and Crous P.W. 2006. Host range of Cercospora apii and C. beticola and description of C. apiicola, a novel species from celery. Mycologia 98: 275–285.
Guillin E.A., de Oliveira L.O., Grijalba P.E., Gottlieb A.M. 2017. Genetic entanglement between Cercospora species associating soybean purple seed stain. Mycological Progress 16: 593–603.
Katoh K. and Standley D.M. 2013. MAFFT multiple sequence alignment software version 7: improvements in performance and usability. Molecular Biology and Evolution 30: 772–780.
Kimber R.B.E. and Paull J.G. 2011. Identification and genetics of resistance to cercospora leaf spot (Cercospora zonata) in faba bean (Vicia faba). Euphytica 177: 419–429.
Maddison W.P. and Maddison D.R. 2011. Mesquite: a modular system for evolutionary analysis.Version 2.75. http://mesquiteproject.org.
Möller E., Bahnweg G., Sandermann H. and Geiger H.H. 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.
Myllys L., Stenroos S. and Thell A. 2002. New genes for phylogenetic studies
of lichenized fungi: glyceraldehyde-3-phosphate dehydrogenase
and beta-tubulin genes. Lichenologist34: 237–246.
Nguanhom J., Cheewangkoon R., Groenewald J.Z., Braun U., To-Anun C. and Crous P.W. 2016. Taxonomy and phylogeny of Cercospora spp. from Northern Thailand. Phytotaxa 233: 27–48.
Nylander J.A.A. 2004. MrModeltest. Version 2.0. Program distributed by the author. Evolutionary Biology Centre, Uppsala University, Uppsala, Sweden.
Quaedvlieg W., Binder M., Groenewald J.Z., Summerell B.A., Carnegie A.J., Burgess T.I. and Crous P.W. 2014. Introducing the consolidated species concept to resolve species in the Teratosphaeriaceae. Persoonia 33: 1–40.
Ronquist F., Teslenko M., Van der Mark P., Ayres D.L., Darling A., Höhna S., Larget B., Liu L., Suchard M.A. and Huelsenbeck J.P. 2012. MrBayes 3.2: Effcient Bayesian phylogenetic inference and model choice across a large model space. Systematic Biology 61: 539–542.
Soares A.P.G., Guillin E.A., Borges L.L., Da Silva A.C., De Almeida Á.M., GrijalbaP.E.,Gottlieb A.M., Bluhm B.H. and De Oliveira L.O. 2015. More Cercospora species infect soybeans across the Americas than meets the eye. PloSOne 10: e0133495.
Tamura K., Stecher G., Peterson D., Filipski A. and Kumar S. 2013.
MEGA6: Molecular Evolutionary Genetics Analysis Version 6.0.
Molecular Biology and Evolution30: 2725–2729.
Vaghefi N., Kikkert J.R., Hay F.S., Carver G.D., Koenick L.B., Bolton M.D., Hanson L.E., Secor G.A. and Pethybridge S.J. 2018. Cryptic diversity, pathogenicity, and evolutionary species boundaries in Cercospora populations associated with Cercospora leaf spot of Beta vulgaris. Fungal Biology 122(4): 264–282.
Weiland J., Eide J., Rivera-Varas V. and Secor G. 2001. Genetic diversity of Cercospora beticola in the US and association of molecular markers with tolerance to the fungicide triphenyltin hydroxide. Phytopathology91: 94.
White T.J., Bruns T. and Taylor J. 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis M.A., Gelfand D.H., Sninsky J.J., et al. (eds), A guide to molecular methods and applications: 315–322. Academic Press, New York.