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مقایسه ناحیه‏ های ژنی ITS، D1/D2 LSU rDNA، بتاتوبولین و RNA پلی‌مراز II در جداسازی گونه‌های Allophoma ،Didymella و Neodidymelliopsis از خانواده Didymellaceae

نوع مقاله : مقاله پژوهشی

نویسندگان

1 استادیار بیماری‌شناسی گیاهی، گروه گیاهپزشکی، دانشکده کشاورزی، دانشگاه شهید چمران اهواز، استان خوزستان، ایران

2 مرکز تحقیقات بیوتکنولوژی و علوم زیستی، دانشگاه شهید چمران اهواز، اهواز، استان خوزستان، ایران

3 استاد بیماری‌شناسی گیاهی، گروه گیاهپزشکی، دانشکده کشاورزی، دانشگاه شهید چمران اهواز، استان خوزستان، ایران

چکیده

در این پژوهش، 12 سویۀ بومی و 80 سویه از گونه‌های شناخته‌شدۀ سه جنس Allophoma،Didymella  و Neodidymelliopsis گزینش شدند و در یک تجزیه‏و‏تحلیل تبارزایشی جهت هم­سنجی ناحیه‏های ژنی ITS، دومین D1 و D2 از زیرواحد بزرگ ژن ریبوزومی (D1/D2 LSU rDNA)، بتاتوبولین و RNA پلی‌مراز در جداسازی گونه‌ها استفاده شدند. ناحیه‏های ژنی سویه‌های بومی با به‌کارگیری DNA استخراج‌شده از زیست‌تودۀ میسیلیومی خشک‌انجمادی شده تکثیر و توالی‌یابی شدند. تجزیه‏و‏تحلیل تبارزایشی با به‌کارگیری الگوریتم درست‌نمائی بیشینه انجام شد. ناحیه‏های D1/D2 LSU rDNA، ITS، rpb2 و tub2 به ترتیب 0، 20، 43 و 46 گونه از 61 گونۀ مربوط به سه جنس موردبررسی را جداسازی کردند. در تجزیه‏و‏تحلیل تبارزایشی ترکیب ناحیه‏های ژنی، همۀ توالی‌های ترکیبی (ITS-tub2، ITS-rpb2، tub2-rpb2، ITS-tub2-rpb2 و ITS-28S-tub2-rpb2) توانستند، نزدیک به 49 گونه از 61 گونۀ موردبررسی را جدا کنند. نتیجه‏ها نشان دادند که برای شناسایی و جداسازی دقیق گونه‌های Allophoma، Didymella و Neodidymelliopsis، توالی‌یابی و تجزیه‏و‏تحلیل تبارزایشی سه ناحیه ITS، tub2 و rpb2 در کنار بررسی‌های ریخت‌شناسی نیازین است. چنانچه به خاطر محدودیت‌های مالی یا زمانی در یک پژوهش، تنها تکثیر و توالی‌یابی یک ناحیه دلخواه باشد، ژنtub2  یا rpb2 بهترین نتایج را ارائه می‌کنند.

کلیدواژه‌ها


عنوان مقاله [English]

A comparison between ITS, D1/D2 LSU rDNA, tub2, and rpb2 regions to delimit Allophoma, Didymella, and Neodidymelliopsis species from the family Didymellaceae

نویسندگان [English]

  • Mehdi Mehrabi-Koushki 1 2
  • Reza Farokhinejad 3
1 Assistant Professor of Plant Pathology, Plant Protection Department, Agriculture Faculty, Shahid Chamran University of Ahvaz, Ahvaz, Khuzestan Province, Iran | Biotechnology and Biological Science Research Center, Shahid Chamran University of Ahvaz, Ahvaz, Iran
2 Assistant Professor of Plant Pathology, Plant Protection Department, Agriculture Faculty, Shahid Chamran University of Ahvaz, Ahvaz, Khuzestan Province, Iran | Biotechnology and Biological Science Research Center, Shahid Chamran University of Ahvaz, Ahvaz, Iran
3 Professor of Plant Pathology, Plant Protection Department, Agriculture Faculty, Shahid Chamran University of Ahvaz, Ahvaz, Khuzestan Province, Iran
چکیده [English]

In this study, 12 native strains and 80 strains from the known species of Allophoma, Didymella, and Neodidymelliopsis were selected. The ITS, D1/D2 LSU rDNA, tub2 and rpb2 regions of the mentioned strains were compared for species delimitation using phylogenetic analysis. The genomic regions of the native strains were amplified using DNA extracted from freeze-dried mycelia and sequenced. The phylogenetic analysis was performed using the maximum likelihood algorithm. The regions of the D1/D2 LSU rDNA, ITS, tub2, and rpb2 delimited 0, 20, 43, and 46 out of 61 species studied, respectively. In the phylogenetic analysis based on combined regions, all datasets (ITS-tub2, ITS-rpb2, tub2-rpb2, ITS-tub2-rpb2, and ITS-28S-tub2-rpb2) could delimit 49 out of the 61 species under survey.Results showed that the sequencing and phylogenetic analysis of the ITS, tub2, and rpb2 regions in combination with morphological studies are necessary for species delimitation of the Allophoma, Didymella and Neodidymelliopsis genera. In a single-locus phylogeny, tub2 or rpb2 genes are the best markers among the genomic regions used.

کلیدواژه‌ها [English]

  • Phylogenetic analysis
  • species identification
  • single-gene and multigene phylogeny
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