وقوع بیماری لکه برگی سرکوسپورایی خربزه در استان گیلان و شناسایی عوامل مرتبط براساس روش های مولکولی و ریخت شناختی

نوع مقاله : مقاله کامل پژوهشی

نویسندگان

1 بخش تحقیقات رستنی‌ها، موسسه تحقیقات گیاهپزشکی کشور، سازمان تحقیقات، آموزش و ترویج کشاورزی، تهران، ایران

2 Agricultural Research, Education and Extension Organization (AREEO), Tehran, Iran

3 استاد/دانشگاه تبریز

4 فارغ التحصیل دکترا

چکیده

طالبی و خربزه (با نام علمی Cucumis melo) از تیره کدوئیان (Cucurbitaceae) یکی از مهمترین گیاهان جالیزی می باشند که در اکثر مناطق ایران کشت می‌شود. طی بازدید از مزارع کشت کدوئیان در استان گیلان، شیوع گسترده‌ای از علایم لکه برگی روی گیاه خربزه مشاهده شد. به منظور شناسایی عوامل دخیل در این بیماری، از برگ‌های دارای علایم لکه برگی، نمونه‌برداری به عمل آمد. طی بررسی میکروسکوپی نمونه ها، مشخص شد، عامل بیماری متعلق به جنس Cercospora است و کشت‌های تک اسپور به صورت مستقیم از لکه های برگی تهیه شدند. در مجموع تعداد 45 جدایه قارچی از گیاهان مختلف به دست آمد که از بین آنها 16 جدایه با استفاده از ترکیب خصوصیات ریخت‌شناختی و توالی پنج ناحیه ژنی ITS، tef1، actA، cmdA و his3 شناسایی شدند. نتایج تجزیه و تحلیل فیلوژنتیکی توالی ها نشان داد، از 16 جدایه قارچی، 4 جدایه به گونه Cercospora sp. G و 12 جدایه به گونه Cercospora cf. flagellaris تعلق داشتند. بر اساس منابع موجود، تحقیق حاضر اولین گزارش از وقوع گونه‌های Cercospora cf. flagellaris و Cercospora sp. G روی خربزه (Cucumis melo) در دنیا و اولین گزارش از وقوع بیماری لکه برگی سرکوسپورایی روی خربزه در ایران است.

کلیدواژه‌ها


عنوان مقاله [English]

Occurrence of Cercospora leaf spot disease on Cucumis melo in Guilan province and characterization of the casual agent(s) using molecular and morphological approaches

نویسندگان [English]

  • Mounes Bakhshi 1
  • Rasoul Zare 2
  • Mahdi Arzanlou 3
  • Abolfazl Narmani 4
1 Department of Botany, Iranian Research Institute of Plant Protection, Agricultural Research, Education and Extension Organization (AREEO), Tehran, Iran
2 Agricultural Research, Education and Extension Organization (AREEO), Tehran, Iran
3 Plant Protection Department, Agriculture Faculty, University of Tabriz, P.O. Box: 5166614766, Tabriz, Iran
4 فارغ التحصیل دکترا
چکیده [English]

Melon (Cucumis melo) which belongs to the family Cucurbitaceae, is one of most important plants, cultivated in different regions of Iran. In a field survey in Guilan province, the widespread distribution of leaf spot symptoms on melon was observed. In order to identify the fungal agents associated with this disease, symptomatic leaves were collected from the field and taken to the laboratory. Single spore cultures derived directly from leaf spots. In total, 45 isolates were obtained of which 16 were identified based on sequences of five genomic loci (ITS, ACT, tef1, cmdA and his3), host plants, and morphological data. The results of five-gene phylogenetic analysis have revealed that, of the 16 isolates, four isolates were identified as Cercospora sp. G and the remaining 12 isolates nested within Cercospora cf. flagellaris. To our knowledge, this is the first report of the Cercospora sp. G and Cercospora cf. flagellaris on Cucumis melo in the world. Moreover, it is the first report of Cercospora leaf spot (CLS) on Cucumis melo in Iran.

کلیدواژه‌ها [English]

  • Cercosporoid fungi
  • Cucurbitaceae
  • Leaf spot
  • multi-gene phylogeny
  • Taxonomy
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.
Arzanlou M., Abeln E.C., Kema G.H., Waalwijk C., Carlier J., Vries I.D., Guzmán M. and Crous P.W. 2007. Molecular diagnostics for the Sigatoka disease complex of banana. Phytopathology 97: 1112–1118.
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., 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. Comprehensive check list of Cercosporoid fungi from Iran. Plant Pathology and Quarantine2: 44–55.
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.
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. and Groenewald J.Z. 2005. Hosts, species and genotypes: opinions versus data. Australasian Plant Pathology 34: 463–470.
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.
Ershad D. 2009. Fungi of Iran. Ministry of Jihad-e-Agriculture. Agricultural Research, Education and Extension Organization, Tehran, Iran.(in Persian)
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)
Fresenius G. 1863. Beiträge zur Mykologie.Vol. 3. Frankfurt: H.L. Brönner.
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.
Ismail H.I., Chan K.W., Mariod A.A. and Ismail M. 2010. Phenolic content and antioxidant activity of cantaloupe (Cucumis melo) methanolic extracts. Food Chemistry 119: 643–647.
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.
Kushalappa A.C., Boivin G. and Brodeur L. 1989. Forecasting incidence thresholds of Cercospora blight in carrots to initiate fungicide application. Plant Disease 73: 979–983
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.
Nylander J.A.A. 2004. MrModeltest. Version 2.0. Program distributed by the author. Evolutionary Biology Centre, Uppsala University, Uppsala, Sweden.
Praveena R. and Naseema A. 2004. Fungi occurring on water hyacinth (Eichhornia crassipes (Mart.) Solms) in Kerala. Journal of Tropical Agriculture 42: 21–23.
Quaedvlieg W., Binder M., Groenewald J.Z., Summerell B.A., Carnegie A.J., BurgessT.I.and Crous P.W. 2014. Introducing the consolidated species concept to resolve species in the Teratosphaeriaceae. Persoonia 33: 1–40.
Ritschel P.S., Lins T.C., Rodrigo Lourenço Tristan R.L., Buso G.S., Buso J.A. and Ferreira M.E. 2004. Development of microsatellite markers from an enriched genomiclibrary for genetic analysis of melon (Cucumis melo L.). BMC Plant Biology 4: 9.
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.
Silva M. and Pereira O.L. 2008. Postharvest Cercospora apii fruit rot disease on Cucurbita maxima (Cucurbitaceae). Australasian Plant Disease Notes3: 21–23.
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.
Tessmann D.J., Charudattan R., Kistler H.C. and Rosskopf E.N. 2001. A molecular characterization of Cercospora species pathogenic to water hyacinth and emendation of C. piaropi. Mycologia 93: 323–334.
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.