Detection and molecular identification of Colletotricum species causing anthracnose disease of some plants in Khuzestan province (Iran)

Document Type : Systematics and Biodiversity of Fungi

Authors

1 MSc Student of plant pathology, Department of Plant Protection, Agriculture Faculty, Shahid Chamran University of Ahvaz, Ahvaz, Iran

2 Professor of plant pathology, Department of Plant Protection, Agriculture Faculty, Shahid Chamran University of Ahvaz, Ahvaz, Iran

3 Assistant Professor of plant pathology, Department of Plant Protection, Agriculture Faculty, Shahid Chamran University of Ahvaz, Ahvaz, Iran; 2 Biotechnology and Bioscience Research Center, Shahid Chamran University of Ahvaz, Ahvaz, Iran

Abstract

Anthracnose is a general term for sunken lesions and blights on stems, leaves and fruits which causes economic damage to agricultural products. This study, conducted to identify Colletotrichum species associated with anthracnose symptoms on some plants in Khuzestan province (Iran) using morphological and molecular methods. During 2016–17, 40 leaf and fruit samples showing anthracnose symptoms (from alfalfa, banana, sour lemon, orange and strawberry) were collected and surveyed for morphological study. Accordingly, 12 isolates of the genus Colletotrichum were obtained. Mycelial biomass of the isolates were produced in PDB and freeze-dried. DNA was extracted using an organic method based on phenol and chloroform. The ITS and partial regions of the 28S-D1/D2 and Tub2 were amplified using universal and specific primers and sequenced. Obtained sequences were compared with index sequences belonging to the type strains in GenBank by BLASTn algorithm and phylogenetic analysis (maximum likelihood algorithm). The species of Colletotrichum gloeosporioides, C. karstii, C. musae, C. nymphaeae, and C. truncatum were identified along with their morphological study. In phylogenetic tree constructed based on the ITS and Tub2 combined dataset, Colletotrichum isolates under study were clustered with type strain of each species and formed monophyletic clades with 96–99% bootstrap support. 
  
  
 

Keywords

Main Subjects

Article Title [Persian]

ردیابی و شناسایی مولکولی گونه ‏های Colletotrichum عامل بیماری آنتراکنوز برخی گیاهان در استان خوزستان

Authors [Persian]

  • کبری حیدری 1
  • رضا فرخی نژاد 2
  • مهدی مهرابی کوشکی 3
1 دانشجوی ارشد بیماری‏شناسی گیاهی، گروه گیاهپزشکی، دانشکده کشاورزی، دانشگاه شهید چمران اهواز، اهواز، ایران
2 استاد گروه بیماری شناسی گیاهی، گروه گیاهپزشکی دانشکده کشاورزی، دانشگاه شهید چمران اهواز، اهواز،
3 استادیار گروه بیماری شناسی گیاهی، گروه گیاهپزشکی، دانشکده کشاورزی، دانشگاه شهید چمران اهواز، اهواز، ایران؛ 2 استادیار گروه بیوتکنولوژی، مرکز تحقیقات بیوتکنولوژی و علوم زیستی، دانشگاه شهید چمران اهواز، اهواز، ایران
Abstract [Persian]

تحقیق حاضر به منظور شناسایی گونه‏ های Colletotrichum همراه علایم آنتراکنوز در برخی گیاهان در استان خوزستان، با استفاده از روش‏ های مبتنی بر ریخت‌شناسی و تجزیه و تحلیل مولکولی انجام شد. طی سال ‏های 96-1395، 40 نمونه برگ و میوه دارای علایم آنتراکنوز از گیاهان توت‏فرنگی، پرتقال، لیمو‏ترش، موز، یونجه و نارنج جمع‏آوری شد که براساس بررسی‌های ریخت‏ شناسی، 12 جدایه آن ‏ها، از جنس Colletotrichum بودند. از این جدایه‏ ها، زیست ‏توده میسیلیومی در محیط PDB تهیه و خشک ‏انجمادی انجام شد. استخراج DNA با استفاده از یک روش آلی مبتنی بر فنل و کلروفرم انجام شد و سپس، ناحیه ITS و بخش ‏هایی از نواحی 28S-D1/D2 و Tub2با استفاده از آغازگرهای عمومی و اختصاصی تکثیر و توالی‌یابی شدند. توالی‏ های حاصله با توالی‏ های مربوط به استرین ‏های تیپ در بانک ژن با استفاده از الگوریتم BLASTn و تجزیه و تحلیل فیلوژنتیکی (الگوریتم درست ‏نمایی بیشینه) مورد مقایسه قرار گرفتند. گونه‏ های Colletotrichum gloeosporioides ،C. karstii، C. musae، C. nymphaeae و C. truncatum شناسایی و ریخت ‏شناسی آن‏ ها بررسی شد. در درخت فیلوژنتیکی مبتنی بر توالی‏ های ترکیبی، دو ناحیه ژنی ITS و Tub2، جدایه ‏های Colletotrichum تحت مطالعه با استرین ‏های معتبر از هر گونه خوشه ‏بندی شدند و کلاد‏هایی معتبر با ارزش بوت‏استراپ 99-96 درصد ایجاد کردند.
 
 
 

Keywords [Persian]

  • ایران
  • تجزیه و تحلیل فیلوژنتیکی
  • توالی‌یابی
  • ریخت ‏شناسی
  • کلتوتریکوم

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History

  • Receive Date: 24 June 2018
  • Revise Date: 05 August 2018
  • Accept Date: 21 August 2018

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