شناسایی و بررسی تنوع‌ژنتیکی استرین‌های بیماری‌زای سودوموناس جداشده از میزبان‌های فلفل، پیاز و سیب‌زمینی در استان‌های آذربایجان‌شرقی و غربی

نوع مقاله : علمی پژوهشی-فارسی

نویسندگان

1 دانشجوی دکتری بیماری شناسی گیاهی، گروه گیاه پزشکی، واحد تبریز، دانشگاه آزاد اسلامی، تبریز، ایران

2 استادیار، گروه گیاه پزشکی، واحد تبریز، دانشگاه آزاد اسلامی، تبریز، ایران

3 استادیار گروه گیاه پزشکی، واحد مهاباد، دانشگاه آزاد اسلامی، مهاباد، ایران

چکیده

بیماری‌ پوسیدگی باکتریایی فلفل، پیاز و سیب‌زمینی یکی از بیماری‌های مهم در استان‌های آذربایجان‌شرقی و غربی است، که شناسایی دقیق عوامل این بیماری‌ و بررسی تنوع ژنتیکی آن‌ها می‌تواند در پیشگیری از وقوع این بیماری‌ها مفید باشد. در تحقیق حاضر سعی می­‌شود علاوه بر مطالعه و شناسایی فنوتیپی-بیوشیمایی و مولکولی، به مطالعه‌ی تنوع ژنتیکی گونه‌های باکتریایی از این میزبان‌های منتخب در استان‌های آذربایجان‌شرقی و غربی پرداخته شود تا اطلاعات کاملی در این خصوص حاصل گردد. بدین منظور در فصول بهار و تابستان سال‌های 1398 و 1399 نمونه‌برداری از گیاهان مزبور صورت گرفت. از کشت نمونه‌های جمع آوری شده، در کل 59 جدایه به عنوان باکتری‌های بیماری‌زا جداسازی شدند. براساس نتایج آزمون‌های فنوتیپی، همه استرین‌ها گرم منفی و کاتالاز مثبت و هوازی اجباری بوده و تولید رنگدانه فلورسنت در محیط کشت KB کردند و به عنوان جنس سودوموناس شناسایی شدند. بیماری‌زایی استرین‌ها پس از گذشت دو هفته روی میزبان سالم، اثبات گردید. تنوع استرین‌ها با مقایسه ویژگی‌های فنوتیپی، انگشت‌نگاری  ژنتیکی حاصل از BOX-PCR و ERIC-PCR با یکدیگر ارزیابی گردید. براساس انگشت‌نگاری  ژنتیکی حاصل از این دو روش، استرین‌های سیب‌زمینی به 5 گروه و استرین‌های جداشده از پیاز و فلفل به 4 گروه تقسیم شدند. این نتایج نشان دهنده تنوع ژنتیکی بالا بین استرین‌ها بود. براساس توالی یابی ژن های 16S rRNA، gyrB و rpoD، استرین‌های جداشده از میزبان‌های سیب‌زمینی، پیاز و فلفل به عنوان گونه Pseudomonas marginalis شناسایی شدند. با بررسی منابع موجود، تحقیق حاضر اولین گزارش از جداسازی، شناسایی و بررسی تنوع ژنتیکی این استرین‌ها از میزبان‌های فلفل، سیب‌زمینی و پیاز در استان‌های آذربایجان‌شرقی و غربی می‌باشد.

کلیدواژه‌ها

عنوان مقاله [English]

Identifying and studying the genetic diversity of pathogenic Pseudomonas strains isolated from pepper, onion, and potato in east and west Azerbaijan provinces

نویسندگان [English]

  • S. Arshad 1
  • S. Nematollahi 2
  • K. Rouhrazi 2
  • N. Kezrinejhad 3
1 Ph.D. Student of Plant Pathology, Department of Plant Protection, Tabriz branch, Islamic Azad University, Tabriz, Iran
2 Associate Professor, Department of Plant Protection, Tabriz branch, Islamic Azad University, Tabriz, Iran
3 Associate Professor, Department of Plant Protection, Mahabad branch, Islamic Azad University, Mahabad, Iran
چکیده [English]

Background and Objectives
The bacterial rot of pepper, onion, and potato is one of the most crucial diseases in the East and West Azerbaijan provinces of Iran. Therefore, accurate identification of the causes of these diseases and the study of their genetic diversity can be beneficial in preventing their occurrence. In the present study, in addition to studying and identifying their morphological and molecular characteristics, the genetic diversity of bacterial species from selected hosts in the provinces of East and West Azerbaijan is studied to obtain comprehensive information.
Material and Methods
Infected potatoes, onions, and peppers with soft rot symptoms were collected from different areas in the East and West Azarbaijan provinces of Iran in the timeframe between 2019 and 2020. Infected samples were cultured on the NA medium. Moreover, purified bacterial colonies were streaked on the Kings’B medium. All isolates that produced fluorescent pigments were identified using biochemical, nutritional, and physiological tests, including LOPAT tests (levan production from sucrose (L), the presence of oxidase (O), the ability to cause rot on potato tubers (P), the presence of arginine dihydrolase (A), and the ability to induce hypersensitivity reaction (HR) on tobacco leaves (T)), nitrate reduction, and the utilization of various carbon sources. All isolates that produced fluorescent pigments were identified as Pseudomonas using phenotypic properties. To assess the genetic diversity within the strains, BOX and ERIC-PCR were analyzed. The UPGMA method was employed to evaluate the similarity matrix and clustering between the strains. For genetic identification, 16S rRNA, gyrB, and ropD genes of strains were amplified and sequenced.
 
Results
A total of 59 isolates were obtained from infected onion, pepper, and potato plants. The bacterial isolates revealed cream colonies with a low convex height, jagged edge, and rugged surface on the nutrient agar medium. All strains produced fluorescent pigments on the King’s B medium and demonstrated Gram-negative and positive reactions to the catalase. In pathogenicity tests, all strains were pathogenic on healthy plants. Moreover, clustering ERIC and BOX-PCR results with UPGMA and Jaccard’s similarity coefficients showed that the strains of potato, pepper, and onion were clustered into five and four main groups, respectively. The gyrB, rpoD, and 16S rRNA nucleotide sequences showed high similarity between the strains and Pseudomonas cichorii.
Discussion
The results showed that Iranian P. marginalis strains are very diverse and are related to the isolation site and the host plant. This high diversity can be due to the long history of cultivating these plants in the provinces of Iran and the high genetic diversity in the bacterial population. Based on previous studies published on Iranian P. marginalis, this is the first report of isolation and identification of P. marginalis, the agent that causes soft rot in pepper, onion, and potato in East and West Azarbaijan provinces.

کلیدواژه‌ها [English]

  • Bacterial rot
  • Phenotypic properties
  • Genetic fingerprinting
  • Genetic diversity

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 © 2022 by the authors. Licensee SCU, Ahvaz, Iran. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution-NonCommercial 4.0 International (CC BY-NC 4.0 license) (http://creativecommons.org/licenses/by-nc/4.0/
گیاه پزشکی
دوره 45، شماره 2
مرداد 1401
صفحه 63-83

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