جداسازی ازتوباکترهای بومی از خاک مناطق مختلف تهران و بررسی اثرتلقیح آنها بر رشد گیاه گوجه‌فرنگی (Solanum lycopersicum L.)

محورهای موضوعی : اکوفیزیولوژی گیاهان زراعی

شقایق گلچین ایرانی ¹ , غلامرضا طاهری سنگسری ² , اکرم سادات طباطبایی بفرویی ³ , محمدجواد اوستا ⁴

1 - دانش‌آموخته گروه زیست شناسی، واحد تهران شرق، دانشگاه آزاد اسلامی، تهران، ایران
2 - عضو هیات علمی گروه زیست شناسی، واحد تهران شرق، دانشگاه آزاد اسلامی، تهران، ایران
3 - استادیار گروه زیست شناسی، واحد تهران شرق، دانشگاه آزاد اسلامی، تهران، ایران
4 - عضو شرکت دانش بنیان رویان تیسان سبز، تهران، ایران

تاریخ دریافت : 1399/07/15 تاریخ پذیرش : 1400/05/31 تاریخ انتشار : 1401/07/01

کلید واژه: بهینه سازی, کود بیولوژیک, ازتوباکتر, جداسازی, گیاه گوجه‌فرنگی, nif H,

چکیده مقاله :

ازتوباکتر، باکتری هوازی، گرم منفی، شیمیوارگانوتروف است که قادر به تثبیت نیتروژن مولکولی به صورت غیر‌همزیست می‌باشد. نقش ازتوباکتر در رشد گیاه به واسطه تولید هورمون‌های محرک رشد، توان حل کنندگی فسفات‌های نامحلول، تثبیت نیتروژن، افزایش تحمل به تنش‌ها و بیوکنترل عوامل بیماری زای گیاهی می‌باشد. این پژوهش با هدف جداسازی و شناسایی ازتوباکتر بومی تثبیت کننده نیتروژن از خاک مناطق مختلف شهر تهران صورت گرفت. همچنین، تاثیر تلقیح جدایه ها بر تقویت رشد گیاه گوجه فرنگی نیز بررسی و در نهایت شرایط رشد جدایه برتر بهینه سازی شد. در این پژوهش، از شش منطقه تهران نمونه ‌برداری انجام و جدایه های ازتوباکتر با استفاده از روش تهیه رقت متوالی از نمونه‌های خاک، جداسازی و با کمک تست‌های بیوشیمیایی متداول تعیین هویت شدند. سپس، ژن nif H، کد کننده آنزیم نیتروژناز، در جدایه ها با استفاده از تکنیک واکنش زنجیره پلیمراز ریل تایم شناسایی و سپس بذرهای گوجه فرنگی با ایزوله ها تلقیح و میزان رشد گیاهچه، شامل طول ساقه و ریشه در طی 34 روز سنجیده شدند. پارامتر‌های دما، pH، میزان هوادهی، منبع کربن و نیتروژن برای جدایه برتر بهینه‌سازی شدند. در این مطالعه 27 جدایه ازتوباکتر تثبیت کننده نیتروژن به دست آمد. تمامی جدایه ها افزایش بارزی ( p<0.05 ) در طول ساقه و ریشه گیاه گوجه فرنگی نسبت به سویه استاندارد و کنترل منفی ایجاد کردند. جدایه شماره 21 بیشترین تاثیر را بر رشد گیاه نشان داد و بهترین شرایط رشد آن، در حضور منبع کربن مانیتول، منبع نیتروژن پپتون، دور گردشی rpm200، دمای 30 درجه سلسیوس و 7 =pH به دست آمد. با توجه به نتایج این مطالعه، جدایه های بومی به دست آمده به ویژه جدایه شماره 21 پس از بررسی های بیشتر دارای قابلیت استفاده به عنوان کود بیولوژیکی هستند.

چکیده انگلیسی:

Azotobacter is an aerobic, gram negative and chemoorganotrophic bacterium, that is able to stabilize molecular nitrogen nonsymbiotically. The role of Azotobacter in plant growth is due to the production of growth-promoting hormones, the ability to dissolve insoluble phosphates, nitrogen fixation, increase stress resistance and biocontrol of plant pathogens. The aim of this study was to isolate and identify the indigenous Azotobacter from the soil of different areas of Tehran. The effect of tomato plant inoculation with isolates on growth promoting was also investigated. Finally, the growth conditions of the superior isolate were optimized.  Azotobacter isolates were obtained from soil samples using serial dilution method and identified by conventional biochemical tests. The nif H gene, encoding nitrogenase enzyme, was identified in isolates using real-time PCR technique. Then the tomato seeds were inoculated with isolates and seedling growth rate including stem and root length were measured during 34 days. The parameters of temperature, pH, aeration rate, and carbon and nitrogen sources were optimized for superior isolate. In this study, 27 isolates were identified as nitrogen fixing Azotobacter. Considering the results, all isolates showed a significant increase (p <0.05) in stem and root length of tomato plants compared to standard strain and negative control. Among them, isolate No. 21 had the greatest effect during 34 days of study and its best growth conditions in the presence of mannitol carbon source, peptone nitrogen source, 200 rpm rotation, 30°C and pH 7were acquired. According to the results of this study, the obtained indigenous isolates particularly isolate No.21 have the potential to be used as biological fertilizer after further investigations.

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