تنوع ژنتیکی جدایه‏ های گونه Colletotrichum fructicola با استفاده از نشانگر مولکولی ISSR و بررسی کارآمدی این تکنیک در تفکیک این گونه از برخی از گونه ‏های جنس Colletotrichum

نوع مقاله : مقاله پژوهشی

نویسندگان

1 گروه گیاهپزشکی، دانشکده کشاورزی، دانشگاه شهید مدنی آذربایجان، تبریز، ایران.

2 مجتمع آموزش عالی شهید باکری میاندوآب، دانشگاه ارومیه.

چکیده

در این پژوهش تنوع ژنتیکی 70 جدایه Colletotrichum fructicola بدست آمده از علایم لکه‌برگی روی انواع گیاهان اهلی و وحشی در مناطق مختلف سه استان شمالی ایران شامل گیلان، مازندران و گلستان با استفاده از نشانگر مولکولی ISSR بررسی شد. تجزیه و تحلیل خوشه ‏ای نتایج حاصل از انگشت‏ نگاری DNA با استفاده از پنج آغازگر ISSR برای 70 جدایه‏ C. fructicola به خوبی توانست روابط ژنتیکی بین جدایه‏ ها را نشان دهد. بر این اساس تعداد 70 جدایه در قالب هشت دودمان همسان ه­ای از هم تفکیک شدند. این نتایج نشان داد تنوع ژنتیکی زیادی بین جدایه‏ های این گونه قارچی وجود دارد. هیچ ارتباط مستقیمی بین گروه‏ های انگشت ‏نگاری DNA و منشاء جغرافیایی و میزبانی جدایه ‏ها مشاهده نشد. به منظور ارزیابی کارآمدی نشانگر مولکولی ISSR در تمایز و تفکیک جدایه ­های C. fructicola از جدایه­ های، برخی گونه‏ های نزدیک و غالب جنس Colletotrichum، انگشت ­نگاری DNA با استفاده از این نشانگر مولکولی برای 89 جدایه شامل 85 جدایه C. fructicola، دو جدایه C. gloeosporioides، یک جدایه C. aenigma و یک جدایه C. karstii انجام شد. بر اساس مقایسه الگوی انگشت‌نگاری DNA و فنوگرام ترسیم شده تعداد 89 جدایه در سطح تشابه ژنتیکی 70 درصد در چهار گروه اصلی (A-D) قرار گرفتند. به طوری که جدایه ‏های دودمان ‏های همسانه­ای A-D به ترتیب به گونه‏های C. karstii، C. fructicola، C. gloeosporioides و C. aenigma تعلق داشتند. برای اطمینان از صحت شناسایی، تعداد 28 جدایه از دودمان‏ های کلونی مختلف به عنوان نماینده برای شناسایی مولکولی بر اساس تعیین توالی ناحیه ژنی بتاتوبولین (TUB2) و فاصله بین ژنی APN2/MAT1 (ApMAT) انتخاب شدند. تجزیه و تحلیل تبارزایی صحت شناسایی و نیز کارآمدی نشانگر مولکولی ISSR به عنوان یک ابزار مفید در گروه ­بندی و تفکیک برخی گونه ‏های غالب جنس Colletotrichum را نشان داد.

کلیدواژه‌ها

عنوان مقاله [English]

Genetic diversity of Colletotrichum fructicola isolates using ISSR molecular marker and efficiency of this technique in differentiating this species from several Colletotrichum species

نویسندگان [English]

  • Somayeh Akbarzadeh 1
  • Alireza Alizadeh 1
  • Abdollah Ahmadpour 2
  • Akbar Shirzad 1
1 Department of Plant Protection, Faculty of Agriculture, Azarbaijan Shahid Madani University, Tbariz, Iran.
2 Higher Education Center Shahid Bakeri Miyandoab, Urmia University. Urmia, Iran.
چکیده [English]

In this study, genetic diversity of 70 isolates of Colletotrichum fructicola obtained from the leaf spot symptoms on a variety of cultivated and wild plants in different regions of three northern provinces of Iran, including Guilan, Mazandaran and Golestan, was investigated using the ISSR molecular marker. Cluster analysis of the results of DNA fingerprinting using five ISSR primers for 70 isolates of C. fructicola cleraly reverlaed genetic diversity among the isolates. Based on this, 70 isolates were differentiated into eight colonal lineages. These results showed that there is a high level of genetic diversity among isolates of this fungal species. No direct correlation between DNA fingerprinting groups and geographical origin and host of the isolates was observed. In order to evaluate the efficiency of ISSR molecular marker in differentiating C. fructicola isolates from isolates of some close and dominant species of the genus Colletotrichum, DNA fingerprinting using this molecular marker for 89 isolates including 85 C. fructicola isolates, two C. gloeosporioides isolates, one isolate for each of C. aenigma and C. karstii was performed. Based on the comparison of the DNA fingerprinting pattern and the drawn phenogram, 89 isolates were placed in four main groups (A-D) at the genetic similarity level of 70%. So that the isolates of colonal lineages A-D belonged to C. karstii, C. fructicola, C. gloeosporioides and C. aenigma species, respectively. To ensure the accuracy of the identification, 28 isolates from different clonal lineages were selected as representatives for molecular identification based on the sequencing of the beta-tubulin (TUB2) gene region and the APN2/MAT1 (ApMAT) intergenic space. Phylogenetic analysis showed the accuracy of identification and the efficiency of ISSR molecular marker as a useful tool in grouping and separating the dominant species of the genus Colletotrichum.

کلیدواژه‌ها [English]

  • DNA fingerprinting
  • Genomic sequencing
  • Molecular markers
  • PCR
  • Taxonom

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پژوهش های کاربردی در گیاهپزشکی
دوره 11، شماره 4
دی 1401
صفحه 43-61

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  • تاریخ دریافت: 08 آذر 1400
  • تاریخ بازنگری: 02 دی 1400
  • تاریخ پذیرش: 12 مرداد 1401

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