جداسازی، شناسایی و بررسی خصوصیات باکتری‌های اندوفیت ارقام ایرانی گندم

نوع مقاله : علمی پژوهشی-فارسی

نویسندگان

1 دانشجوی دکتری بیماری شناسی گیاهی، دانشکده کشاورزی، دانشگاه فردوسی مشهد، مشهد، ایران

2 دانشیار گروه گیاه‌پزشکی، دانشکده کشاورزی، دانشگاه فردوسی مشهد، مشهد، ایران

3 استاد گروه گیاه‌پزشکی، دانشکده کشاورزی، دانشگاه فردوسی مشهد، مشهد، ایران

چکیده

گندم یکی از محصولات زراعی مهم در سراسر جهان است. امروزه کشاورزی سنتی وابسته به کنترل شیمیایی می‌باشد که می‌تواند اکوسیستم‌ها را نابود نماید. هدف از این تحقیق، شناسایی باکتری‌های اندوفیت عامل مهار زیستی و افزایش دهنده رشد گندم بود. در این پژوهش 464 باکتری از بذر‌ها، اندام‌های هوایی و ریشه‌های 14 رقم ایرانی گندم (ارگ، بهار، بهاران، بهرنگ، چمران، چمران2، دنا، فلات، حیدری، پارسی، پیشگام، سایون، سیروان و سیوند) جداسازی شدند و علیه دو بیمارگر مهم Fusarium graminearum و Xanthomonas translucens مورد ارزیابی قرار گرفتند. حداکثر شاخص بازدارندگی از رشد قارچ در کشت متقابل، 66/86 درصد مربوط به جدایه CB2 و حداکثر هاله جلوگیری از رشد باکتری، 33/49 میلی‌متر در جدایه‌هایPaR28  و PiR5 مشاهده شد. دوازده جدایه آنتاگونیست قوی که جوانه‌زنی بذر و رشد گیاهچه را نیز در آزمایشگاه بالا بردند، برای بررسی بیشتر خصوصیاتشان انتخاب شدند. همگی قادر به تولید پروتئاز، پکتیناز، سلولاز، آمیلاز، IAA، آمونیاک، سیدروفور و قابل حل نمودن فسفات بودند. شاخص‌های رشدی گیاه از جمله طول، وزن تر و خشک اندام‌های هوایی و ریشه‌ها، در گیاهچه‌های گندم تیمار شده با اندوفیت‌ها، افزایش یافتند. پنج جدایه نماینده با توالی‌یابی ژن 16S rRNA به عنوانsp.  Bacillus (MT258405) و sp. Paenibacillus (MT258403، MT258407، MT258404 و MT258406) شناسایی شدند. شباهت جدایه Bacillus با گونه‌های B. subtilis، B. amyloliquefaciens و B. velezensis و شباهت جدایه‌های Paenibacillus با P. jamilae، P. polymyxa و P. peoriae بیش از 99 درصد بود. نتایج بیانگر وجود دو جنس غالب باکتری اندوفیت همراه گندم بود که توانایی مهارزیستی و تحریک رشد گیاه را داشتند.

کلیدواژه‌ها

عنوان مقاله [English]

Isolation, identification, and characterization of bacterial endophytes in Iranian wheat cultivars

نویسندگان [English]

  • E. Taheri 1
  • S. Tarighi 2
  • P. Taheri 3
1 PhD student of plant pathology, Department of Plant Protection, College of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran
2 Associate Professor, Department of Plant Protection, College of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran
3 Professor, Department of Plant Protection, College of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran
چکیده [English]

Background and Objectives
Wheat (Triticum aestivum L.) is one of the most important crops worldwide, responsible for a large portion of the human diet. In light of the increasing population, increased demand of 20 million tons by 2025 has been predicted by the Iranian Ministry of Agriculture. However, Fusarium graminearum and Xanthomonas translucens are among the most destructive diseases that lead to high yield loss. The present study aimed to isolate and evaluate the antagonistic plant growth-promoting endophytic bacteria in Iranian wheat cultivars.
Materials and Methods
In this study, endophytic bacteria were isolated from different tissues (e.g., seeds, roots, and shoots) of 14 wheat cultivars: Arg, Bahar, Baharan, Behrang, Chamran, Chamran 2, Dena, Falat, Heidari, Parsi, Pishgam, Sayonz, Sirvan, and Sivand. Bacterial strains were cultured in different media (i.e., NA, LBA, and TSA 10%). Then, their antifungal and antibacterial activity against F. graminearum and X. translucens were analyzed by dual culture assays. Moreover, the effect of isolates on seed germination was evaluated in vitro. In addition, extracellular enzyme production and plant growth-promoting properties were assessed. Furthermore, the effect of endophytes on plant growth was evaluated in greenhouse trials, and finally, the selected isolates were identified by 16S rRNA gene sequencing using two sets of universal primers (rD1, fD1, and 27F, 1492R).
Results
In this study, a total of 464 endophytic bacteria were isolated from wheat surface-sterilized tissues. Forty-four isolates successfully suppressed both Fusarium and Xanthomonas in vitro. The highest percentage of the fungal growth inhibition was evaluated at 86.66% (CB2), and the highest clear halo around isolates, which indicated the antibacterial activity, was 49.33% (PaR28, PiR5). Among the isolated bacteria, twelve were capable of increasing seed germination in vitro by at least 10% more than untreated seeds and could increase shoot and root length, which were selected for further characterization. The clear halos, which indicated enzyme production, were calculated as follows: protease 0.36 to 2.53, cellulase 1.03 to 1.75, pectinase 1.06 to 2.65, and amylase 0.43 to 2.1 cm. The diameter of siderophore production on CAS agar medium varied from 0.21 to 1.46 cm. Moreover, IAA was between 7.05 and 14.14 µg/mL, ammonia production varied from 0.29 to 34.49 mg/mL, and phosphate solubilization was between 6.09 and 14.95 µg/mL. Greenhouse experiments indicated that the biomass was significantly increased in plants treated with endophytes. In addition, DNA sequencing using the 16S rRNA gene showed that the isolate CB2 had 99.06% homology with B. subtilis, B. velezensis, and B. amyloliquefaciens. Plus, the isolates PaR28, SeR8, BaR15, and BR20 indicated a homology of more than 99% with P. jamilae, P. polymyxa, and P. peoriae. The sequence of the mentioned isolates has been deposited in the GenBank under accession numbers MT258405 (CB2), MT258403 (BaR15), MT258407 (SeR8), MT258404 (BR20), and MT258406 (PaR28).
Discussion
The present study is the first report on the endophytic population in mentioned wheat cultivars and an indication of their significance in the biocontrol of X. translucens. As isolates identified in this study have several desirable characteristics, a better-informed knowledge of them can be expected to have promising applications as biofertilizers and biopesticides.

کلیدواژه‌ها [English]

  • Biofertilizers and biopesticides
  • Bacillus sp
  • Fusarium graminearum
  • Paenibacillus sp
  • Xanthomonas translucens

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گیاه پزشکی
دوره 45، شماره 2
مرداد 1401
صفحه 91-108

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