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Detection and molecular identification of Colletotricum species causing anthracnose disease of some plants in Khuzestan province (Iran) | ||
| Rostaniha | ||
| مقاله 3، دوره 19، شماره 1 - شماره پیاپی 55، دی 2018، صفحه 19-32 اصل مقاله (751.82 K) | ||
| نوع مقاله: Systematics and Biodiversity of Fungi | ||
| شناسه دیجیتال (DOI): 10.22092/botany.2018.122372.1111 | ||
| نویسندگان | ||
| Kobra Heidari1؛ Reza Farokhinejad2؛ Mehdi Mehrabi-Koushki* 3 | ||
| 1MSc Student of plant pathology, Department of Plant Protection, Agriculture Faculty, Shahid Chamran University of Ahvaz, Ahvaz, Iran | ||
| 2Professor of plant pathology, Department of Plant Protection, Agriculture Faculty, Shahid Chamran University of Ahvaz, Ahvaz, Iran | ||
| 3Assistant 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 | ||
| چکیده | ||
| 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. | ||
| کلیدواژه‌ها | ||
| Morphology؛ Phylogenetic analysis؛ sequences؛ strains | ||
| عنوان مقاله [English] | ||
| ردیابی و شناسایی مولکولی گونه های Colletotrichum عامل بیماری آنتراکنوز برخی گیاهان در استان خوزستان | ||
| نویسندگان [English] | ||
| کبری حیدری1؛ رضا فرخی نژاد2؛ مهدی مهرابی کوشکی3 | ||
| 1دانشجوی ارشد بیماری‏شناسی گیاهی، گروه گیاهپزشکی، دانشکده کشاورزی، دانشگاه شهید چمران اهواز، اهواز، ایران | ||
| 2استاد گروه بیماری شناسی گیاهی، گروه گیاهپزشکی دانشکده کشاورزی، دانشگاه شهید چمران اهواز، اهواز، | ||
| 3استادیار 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 | ||
| چکیده [English] | ||
| 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. | ||
| کلیدواژه‌ها [English] | ||
| ایران, تجزیه و تحلیل فیلوژنتیکی, توالی‌یابی, ریخت شناسی, کلتوتریکوم | ||
| مراجع | ||
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