ترکیب جمعیتی باکتری های اندوفیت بقولات وحشی به عنوان عوامل بالقوه کنترل زیستی بیماری‌های بقولات زراعی ‌ (مطالعه موردی: جنگل زاگرس کرمانشاه)

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانش‌آموخته مقطع دکتری، دانشکده علوم کشاورزی و صنایع غذایی، واحد علوم و تحقیقات دانشگاه آزاد اسلامی، تهران، ایران.

2 دانشیار، دانشکده علوم کشاورزی و صنایع غذایی، واحد علوم و تحقیقات دانشگاه آزاد اسلامی، تهران، ایران

3 استادیار، پژوهشکده زیست‌فناوری صنعت و محیط‌زیست، پژوهشگاه ملی مهندسی ژنتیک و زیست‌فناوری، تهران، ایران

4 محقق گروه تنوع زیستی، مؤسسه بوم زیستا، ونکوور، کانادا

5 استادیار، گروه بیوتکنولوژی و به‌نژادی، دانشکده علوم کشاورزی و صنایع غذیی، واحد علوم و تحقیقات دانشگاه آزاد اسلامی، تهران، ایران

چکیده

اکوسیستم‌های جنگلی مخازن اصلی تنوع زیستی جهانی هستند. میکروارگانیسم‌ها نقش اساسی در چرخه‌های بیوشیمیایی این اکوسیستم‌ها دارند و برای محافظت از گیاهان و تحریک رشد آن‌ها مورد استفاده قرار می‌گیرند از اینرو، شناخت آن ها حائز اهمیت است. بدلیل محدودیت‌های ناشی از اعمال روش‌های پیشین در جداسازی و شناسایی اندوفیت، پتانسیل و عملکرد آنها بدرستی شناخته نشده است. در این پژوهش، از روش غیر وابسته به کشت برای بررسی جامعه باکتریایی اندوفیتی برخی بقولات وحشی استفاده شده است.از جنگل‌های زاگرس کرمانشاه از گیاهان Astragalus ovinus و Vicia lutea نمونه‌برداری شد. پس از استخراج DNA تام از ریشه و برگ‌های گیاهان با کیت استخراج و تکثیر بخشی از ژن 16S rDNA باکتریایی، توالی‌یابی به کمک توالی‌یابی نسل جدید (NGS) و با استفاده از پلتفرم Illumina MiSeq انجام گردید. آنالیز توالی‌ها نشان داد فراوانی نسبی، اندوفیت‌های جداسازی شده متعلق به هفت راسته Rhizobiales، Xanthomonadales، Sphingomonadales، Pseudomonadales، Chitinophagales، Enterobacteriales و Betaproteobacteriales بودند که در مورد اخیر، این راسته به سه رده Gammaproteobacteria، Alphaproteobacteria و Bacteroidia و دو شاخه Proteobacteria و Bacteroidetes تعلق داشت. نتایج نشان داد باکتری‌های شاخه Proteobacteria، اندوفیت‌های باکتریایی غالب بوده و بیش از 99 % کل جمعیت اندوفیت در گونه V. lutea و 68 %در گونه A. ovinus را به خود اختصاص دادند. باکتری‌های مذکور با بیشترین %واحدهای تاکسونومیکی ((OTUs، دارای بیشترین پتانسیل استفاده در کنترل بیولوژیک بیماری‌های بقولات زراعی هستند. این مطالعه برای اولین بار، تصویر روشنی از جمعیت، چیرگی و تنوع اندوفیت‌های باکتریایی گیاهان بقولات وحشی در جنگل‌های زاگرس را با استفاده از آنالیز متاژنومی ارائه می دهد.

کلیدواژه‌ها

عنوان مقاله [English]

Composition of endophytic bacterial communities of wild legumes as potential biological control agents of crop legumes diseases (Case study: Kermanshah Zagros forest)

نویسندگان [English]

  • Sh. Rostami 1
  • nader hasanzadeh 2
  • S. Rajaei 3
  • A. Golnaraghi 4
  • R. Azizinezhad 5
1 1- Department of Plant Protection, Faculty of Agricultural Sciences and Food Industries, Science and Research Branch, Islamic Azad University, Tehran, Iran
2 Department of Plant Protection, Faculty of Agricultural Sciences and Food Industries, Science and Research Branch, Islamic Azad University, Tehran, Iran
3 Department of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran
4 Department of Biodiversity, BoomZista Institute, Vancouver, British Columbia, Canada
5 Department of Plant Breeding and Biotechnology, Faculty of Agricultural Sciences and Food Industries, Science and Research Branch, Islamic Azad University, Tehran, Iran
چکیده [English]

Forest ecosystems are the main reservoirs of global biodiversity. Microorganisms play a key role in the biochemical cycles of these ecosystems and are used to protect plants and stimulate their growth. Due to the limitations in the application of classic methods in the isolation and identification of endophytic microorganisms, their potentials and functions have yet to be properly understood. In this study, a culture-independent method was used to study the endophytic bacterial communities of some wild legumes. To this end, the Zagros forests of Kermanshah were surveyed and two varieties of Astragalus ovinus and Vicia lutea were sampled. Total DNAs from different parts of the plant samples were extracted and the bacterial 16S rDNA genes were subsequently amplified. Sequencing analysis using the Illumina MiSeq platform showed the most abundant endophytic bacteria belonged to seven orders of Rhizobiales, Xanthomonadales, Sphingomonadales, Pseudomonadales, Chitinophagales, Enterobacteriales, Betaproteobacteriales. The latter were comprised to three classes of Gammaproteobacteria, Alphaproteobacteria and Bacteroidia, and two phyla of Proteobacteria and Bacteroidetes. Our findings showed that members of Proteobacteria were the dominant bacterial endophytes and accounted for more than 99% and 68% of the total population of endophytic bacteria in V. lutea and A. ovinus, respectively. These bacteria, with the highest percentage of taxonomic units (OTUs), have the highest potential to use as biological control agents of crop legumes diseases. This study provides a clear picture of the population, dominance and diversity of bacterial endophytes of wild legumes in the Zagros forests using metagenomic analysis.

کلیدواژه‌ها [English]

  • Second generation sequencing
  • microbial communities
  • plant endophytes
  • biological control

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  • تاریخ دریافت: 30 اردیبهشت 1400
  • تاریخ بازنگری: 28 خرداد 1400
  • تاریخ پذیرش: 20 تیر 1400

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