شناسایی مولکولی برخی از ریزجانداران محرک رشد گیاهی و بررسی تأثیر آن‌ها بر برخی از شاخص‌های رشدی گندم

نوع مقاله : مقاله کامل علمی پژوهشی

نویسندگان

1 دانشیار مؤسسه تحقیقات خاک و آب، سازمان تحقیقات آموزش و ترویج کشاورزی، کرج، ایران،

2 استادیار مؤسسه تحقیقات خاک و آب،سازمان تحقیقات آموزش و ترویج کشاورزی، کرج، ایران

3 دانشیار مؤسسه تحقیقات خاک و آب، سازمان تحقیقات آموزش و ترویج، کشاورزی، کرج، ایران،

4 دانش آموخته کارشناسی ارشد گروه علوم خاک، دانشگاه صنعتی اصفهان

چکیده

سابقه و هدف: گندم یکی از با اهمیت‌ترین محصولات غذایی است. در کشاورزی نوین بدلیل افزایش جمعیت و تأثیر منفی استفاده از کودهای شیمیایی از قبیل آلودگی‌های زیست محیطی و به خطر افتادن سلامتی انسان به کارگیری روش‌های مناسب از قبیل تولید کودهای زیستی که چنین خطراتی را نداشته باشد ضروری بنظر می‌رسد. قارچ‌ها و باکتری‌های افزایش دهنده رشد گیاه استفاده شده در کودهای زیستی ریزجانداران مفیدی هستند که با سازوکار‌های متفاوت به صورت مستقیم و غیر مستقیم باعث افزایش رشد، عملکرد و مقاومت گیاهان در برابر تنش‌های زنده و غیر زنده می‌شوند. با اینکه در سال‌های اخیر کودهای زیستی مورد توجه بیشتری قرار گرفته‌اند اما عدم شناسایی دقیق ریزجانداران استفاده شده در تولید این نوع از کودها یکی از مشکلات کودهای زیستی در ایران است. عدم شناسایی دقیق گونه ممکن است بدلیل نقصان روش‌های ریخت‌شناسی و یا بیوشیمیایی در قارچ‌ها و باکتری‌ها باشد. بنابر این هدف از انجام این تحقیق، شناسایی مولکولی جدایه‌های قارچی و باکتری و همچنین بررسی تأثیر این جدایه‌ها به تنهایی و در ترکیب با قارچ میکوریز آربوسکولار روی برخی از خصوصیات رشدی گیاه گندم بود.

مواد و روش‌ها: پس از تهیه جدایه‌های قارچی و باکتریایی از موسسه تحقیقات خاک و آّب، که تحت عنوان ریزجانداران محرک رشد گیاهی شناخته شده‌اند، شناسایی دقیق هر یک از جدایه‌ها به روش ریخت‌شناسی و یا مولکولی صورت پذیرفت. استخراج DNA ژنومی در قارچ میکوریز آربوسکولار به روش تک اسپور و در سایر جدایه‌های باکتری و قارچی به روش مبتنی بر CTAB انجام گرفت. تکثیر ناحیه ژنی 16S rDNA در جدایه‌های باکتری، نواحی TEF-1α و ITS1-5.8S-ITS2 در گونه‌های تریکودرما، ناحیه ITS1-5.8S-ITS2 در Serendipita indica و ناحیه SSU-ITS1-5.8S-ITS2-LSU به‌روش آشیانه‌ای در قارچ میکوریز آربوسکولار صورت گرفت. طی یک آزمون گلخانه‌ای، تأثیر جدایه‌های قارچی و باکتریایی به صورت مجزا و ترکیبی همراه و عدم همراه با قارچ میکوریز آربوسکولار روی شاخص‌های رشدی گیاه گندم از جمله ارتفاع گیاه، وزن خشک هوایی و ریشه، ارتفاع و وزن خوشه و وزن دانه بررسی شدند.


یافته‌ها: بر اساس بررسی‏های ریخت‏شناسی و مولکولی، سه گونه از جنس باسیلوس شامل Bacillus velezensis، B. pumilus وB. subtilis و سه گونه از جنس تریکودرما شامل Trichoderma atroviride، T. longibrachiatum و T. harzianum و گونه‌های S. indica و Rhizophagus irregularis شناسایی شدند.از نظر تأثیر تیمارهای مختلف روی وزن خشک هوایی و وزن خوشه، تیمارهای T. harzianum + R. irregularis و T. longibrachiatum بیشترین وزن را نشان دادند. تیمارهای T. longibrachiatum + R. irregularis و T. atroviride بیشترین تأثیر را روی ارتفاع گیاه نشان دادند. گلدان‌های تیمار شده باT. harzianum + R. irregularis، T. harzianum، T. atroviride و S. indica + R. irregularis در مقایسه با شاهد بیشترین افزایش وزن دانه را نشان دادند.

نتیجه‌گیری: تیمارهای قارچی T. harzianum + R. irregularis و گونه‌های Trichoderma باعث افزایش معنی‌دار ویژگی‌های رشدی گندم گردید. بر اساس نتایج این تحقیق استفاده از زاد‌مایه مناسب ترکیبی و تهیه فرمولاسیون‌های اولیه برای کاربرد در آزمایش‌های مزرعه‌ای جهت افزایش دسترسی مواد غذایی و بهبود رشد گیاه گندم پیشنهاد می‌شود.

کلیدواژه‌ها


عنوان مقاله [English]

Molecular identification of some plant growth promoting microorganisms and their evaluation on growth characteristic of wheat

نویسندگان [English]

  • Farhad Rejali 1
  • Hossein Kari Dolatabad 2
  • Hadi Asadi Rahmani 3
  • Fahimeh Fazlikhani 4
1 Associate professor, Soil and Water Research Institute
2 Assistant professor, Soil and Water Research Institute, Agricultural Research, Education and Extension Organization (AREEO)
3 Associate professor, Soil and Water Research Institute, Agricultural Research, Education and Extension Organization (AREEO)
4 Former M.Sc. student, Department of Soil Science, College of Agriculture, Isfahan University of Technology, Isfahan, Iran
چکیده [English]

Background and objectives: Wheat is one of the most important food crops. In modern agriculture, Due to the increasing human population and the detrimental effects of chemical fertilizer such as environmental pollution and concerns about human health, adapting suitable alternatives like production of bio-fertilizers which have none of these dangerous effects would be necessary. Plant growth-promoting fungi and bacteria used in bio-fertilizers are the beneficial microorganisms that can enhance plant growth, yield and resistance to biotic and abiotic stresses directly or indirectly through a wide variety of mechanisms. Although bio-fertilizers have become more noticeable in recent years, the lack of accurate identification of microorganisms used in the production of these fertilizers is one of the problems of bio-fertilizers in Iran. Failure to accurate identification of species is due to deficiencies in morphological or biochemical methods in fungi and bacteria. Therefore, the purpose of this study was to identify of fungal and bacterial isolates molecularly and also to investigate the effect of these isolates alone and in combination with arbuscular mycorrhizal fungus on some growth characteristics of the wheat plant.

Materials and methods: After the preparation of fungal and bacterial isolates from the Soil and Water Research Institute, which are known as plant growth promoting microorganisms, accurate identification of each isolate was performed based on morphological and molecular methods. Extraction of genomic DNA in arbuscular mycorrhizal fungus was done by single-spore method and in other bacterial and fungal isolates by CTAB method. Amplification of 16S rDNA region in bacterial isolates, TEF-1α and ITS1-5.8S-ITS2 regions in Trichoderma species, ITS1-5.8S-ITS2 region in Serendipita indica and SSU-ITS1-5.8S-ITS2- LSU region by nested PCR method in the arbuscular mycorrhizal fungus was performed. To investigate the effect of fungal and bacterial isolates alone or in combination with arbuscular mycorrhizal fungus in a greenhouse, their effect on wheat growth indices such as plant height, shoot and root dry weight, head height and weight and dry weights of seeds were measured.

Results: Based on morphological and molecular investigation, three species of Bacillus including Bacillus velezensis, B. pumilus, and B. subtilis and three species of Trichoderma including Trichoderma atroviride, T. longibrachiatum, and T. harzianum and S. indica and Rhizophagus irregularis were identified. T. harzianum + R. irregularis and T. longibrachiatum showed the maximum dry weight and head weight. T. longibrachiatum + R. irregularis and T. atroviride showed the highest effect on plant height. Pots treated with T. harzianum + R. irregularis, T. harzianum, T. atroviride, and S. indica+ R. irregularis showed the highest weight of seeds compared to control plants.

Conclusion: The result showed that the effect of fungal treatments such as T. harzianum + R. irregularis and Trichoderma spp. on growth characteristics of wheat was significant. According to the results of this study, the use of an appropriate combination of inoculants and the preparation of primary formulations for use in farm experiments is proposed to increase the availability of nutrients and improve the growth of wheat plants.

Keywords: Arbuscular mycorrhizal fungus, Bacillus, Plant growth promoting fungi, Trichoderma

کلیدواژه‌ها [English]

  • Arbuscular mycorrhizal fungus
  • Bacillus
  • Plant growth promoting fungi
  • Trichoderma
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