جداسازی، شناسایی مولکولی و ارزیابی ویژگی‌های محرک رشدی باکتری‌های تشکیل دهنده بیوفیلم از ریزوسفر گیاهان غیرزراعی در شمال غرب ایران

نوع مقاله : مقاله پژوهشی

نویسندگان

1 گروه علوم و مهندسی خاک دانشگاه مراغه

2 گروه علوم و مهندسی خاک، دانشکده کشاورزی، دانشگاه تبریز

3 علوم و مهندسی خاک، دانشکده کشاورزی، دانشگاه تبریز

4 گروه زراعت و اصلاح نباتات، دانشکده کشاورزی، دانشگاه مراغه

چکیده

باکتری­های ریزوسفری تشکیل دهنده بیوفیلم با ویژگی­های مطلوب محرک رشدی می­توانند تأثیر زیادی بر تولید گیاهان زراعی در شرایط تنش داشته باشند. بنابراین به‌منظور مطالعه این باکتری‌ها، 50 نمونه­ ریزوسفری از ریشه­ گیاهان وحشی گندمیان تهیه و پس از تهیه سوسپانسون در شرایط آزمایشگاهی روی محیط کشت TSA کشت و باکتری‌ها خالص‌سازی شدند. نتایج نشان داد که بیش از 90 درصد باکتری­های به‌دست آمده توانایی تشکیل بیوفیلم را دارا بوده و می‌توانند در پنج گروه روشناور، چسبندگی زیاد، چسبندگی متوسط، چسبندگی ضعیف و بدون قابلیت چسبیدن تقسیم­بندی شوند. ارزیابی تنوع ژنتیکی باکتری­های مؤثر جداسازی شده نشان داد که در شرایط این مطالعه به شش جنس مختلف باکتریایی تعلق دارند، اما بالغ بر 80 درصد آن‌ها از جنس باسیلوس بودند. نتایج مقاومت به خشکی روی 68 جدایه از 130 جدایه جداسازی شده با قدرت بالای تشکیل بیوفیلم با روش سوربیتول مشخص نمود که در پتانسیل ماتریک 25- بار تمامی جدایه­ها قادر به فعالیت و رشد بودند. از نظر ویژگی­های محرک رشدی تمامی جدایه­ها قادر به تولید هورمون اکسین در محدوده شش تا 65 میکروگرم بر میلی‌لیتر بوده، 96 درصد از جدایه‌ها قادر به تخریب 1-آمینو سیکلوپروپان کربوکسیلیک اسید و تولید آلفا کتو بوتیریک اسید تا 14/2 میکرومول در 36 ساعت بودند. توانایی انحلال فسفر بین 2/1 تا 251 میلی­گرم بر لیتر و قدرت آزادسازی پتاسیم از کانی مسکوویت در آن‌ها بین 90 تا 250 میلی­گرم بر لیتر متغیر بود. در بین باکتری­های مورد مطالعه فقط 41/4 درصد دارای توان تولید سیدروفور بودند. به‌نظر می­رسد که مجموعه باکتریایی خوبی در این مطالعه شناسایی شده­اند که می­توانند به‌عنوان محرک رشد گیاهان در آزمایش‌های گلخانه‌ای و گلدانی و سپس مزرعه­ای در شرایط تنش کم­آبی مورد استفاده قرار گیرند.

کلیدواژه‌ها

عنوان مقاله [English]

Isolation, Molecular Identification, and Assessing Plant Growth Promoting Activities of Biofilm Forming Bacteria from Gramineae Rhizosphere in North West of Iran

نویسندگان [English]

  • Esmaeil Karimi 1
  • Naser Aliasgharzad 2
  • Mohammad Reza Neyshabouri 3
  • Ezatallah Esfandiari Esfandiari 4
1 Soil Science Department, University of MAragheh
2 Soil Science department, faculty of agriculture, University of Tabriz
3 Soil science department, faculty of agriculture, university of Tabriz
4 agronomy department, faculty of agriculture, university of Margheh.
چکیده [English]

Plant-associated rhizospheric bacteria which form biofilms have higher positive effects on plants in fluctuating environments. Thus, to study of these bacteria, 50 rhizospheric samples were taken from grass geramineae roots. After preparing the suspension, 100 microlitere was cultured on TSA medium and then pure culture of bacteria was prepared. Resulted showed that up to 90% of these isolates were able to form biofilm, so that they could be categorized in five groups as: pellicle, very adherent, moderately adherent, weakly adherent and non-adherent. 16S rDNA analysis revealed six different genus of bacteria. Bacillus are the predominant group (80%) of robust biofilm forming in the grass geramineae rhizosphere. The water deficit tolerance of isolates was assessed by rising concentration of sorbitol (1, 10, 20 and 30 %, w/v). All isolates were able to grow in -25 bar matric potential. Isolates showed good potential to producing auxins ranging from 6 - 65 µg mL-1, ACC-deaminase activity (up to 2.14 µmol α-ketobutyrate in 36 h), Ca3(PO4)2 solublization ranging from 1.2 – 251 mg L-1 and K releasing ability 90 to 250 mg L-1. Only three bacteria were able to produce siderophore. It seems that these isolates can be suitable candidates for doing some greeanhaous or pot experiment subjected to protecting plants against water deficit stress. Moreover, they can act as plant growth promoting bacteria as well.

کلیدواژه‌ها [English]

  • 16S rDNA
  • ACC-deaminase
  • Bacillus
  • K releasing
  • Water deficit stress

مراجع

Abd El-Daim I.A., 2015. Use of rhizobacteria for the alleviation of plant stress PhD. Thesis. Swedish University of Agricultural Sciences, Uppsala
Alami Y., Achouak W., Marol C., and Heulin T. 2000. Rhizosphere soil aggregation and plant growth promotion of sunflowers by an exopolysaccharide-producing Rhizobium sp. strain isolated from sunflower roots. Applied Environmental Microbiology, 66: 3393-8.
Archana D.S., Nandish M.S., Savlagi V.P. and Alagawadi A.R. 2013. Characterization of potassium solubilizing bacteria (KSB) from rhizospher soil. Bioinfolet, 10: 248-257.
Bent E., Tuzan S., Chanway C.P. and Enebak S. 2000. Alteration in plant growth and in root hormone levels of lodgeple pines inoculated with rhizobacteria. Canadian Journal of Microbiology, 47: 793-800.
Chang W.S., van de Mortel M., Nielsen L., Nino de Guzman G., and Li X. 2007. Alginate production by Pseudomonas putida creates a hydrated microenvironment and contributes to biofilm architecture and stress tolerance under water limiting conditions. Journal of Bacteriology, 189: 8290–8299.
Crowley D.E., Wang Y.C., Reid C.P.P., and Szaniszlo P.J. 1991. Mechanism of iron acquisition from siderophores by microorganisms and plants. Plant and Soil, 130: 179-198.
E.E.A., 2011. Europe’s Environmentan Assessment of Assessments. European Environment Agency, Copenhagen.
Fleury D., Jefferies S., Kuchel H. and Langridge P. 2010. Genetic and genomic tools to improve drought tolerance in wheat. Journal of Experimental Botany, 61: 3211–3222.
Foley J.A., Ramankutty N., Brauman K.A., Cassidy E.S., and Gerber J.S. 2011. Solutions for a cultivated planet. Nature, 478: 337–342.
Glick B.R, 2014. Bacteria with ACC deaminase can promote plant growth and help to feed the world. Microbiological Research, 169: 30–39.
Gupta A., and Gopal M. 2008. Siderophore production by plant growth promoting rhizobacteria. Indian Journal of Agricultral Research, 42: 153 -156.
Ipek M., and Estiken A. 2017. The action of PGPR on micronutrient availability on soil and plant under calacareous soil condition: An evalution over Fe nutrition. Plant and Microb Interaction in Agro-Ecological, 5: 81-100.
Jayasinghearachchi H.S., and Seneviratne G. 2006. Fungal solubilization of rock phosphate is enhanced by forming fungal–rhizobial biofilms. Soil Biology and Biochemistry, 38: 405-408.
Karimi E., Aliasgharzad N., Neishabouri M. R., and Esfandiari E. 2017. Effect of biofilm PGPRs in alleviation of terminal growth stage water shortage on wheat’s component yield and root. Iranian Journal of Dryland Agriculture, 6: 89-102. (In Persian)  
Kasim W.A., Gaafar RM, Abou-Ali R.M, Omar M.N., and Hewait H.M. 2016. Effect of biofilm forming plant growth promoting rhizobacteria on salinity tolerance in barley. Annals of Agricultural Science, 61: 217–227.
Kavamura V.N., Santos S.N.,   Silva J.L., Parma M.M., Avila L.A., Visconti A., Zucchi T.D., Taketani R.G., Andreote F.D., Melo I.S. 2013. Screening of Brazilian cacti rhizobacteria for plant growth promotion under drought. Microbiological Research, 168: 183–191.
Khan L.A., Lee I.J. 2016. Indol acetic acid and ACC deaminase from endophytic bacteria improves the growth of Solanum lycopersicum. Electronic Journal of Biotechnology, 21: 58-64.
Kumar J.C., and Saraf M. 2015. Plant growth promoting rhizobacteria (PGPR): A review. Journal of Agricultural Research and Development, 5:0108-0119:
Morikawa M., Kagihiro S., Haruki M., Takano K., Branda S., Kolter R., and Kanaya S. 2006: Biofilm formation by a Bacillus subtilis strain that produces c-polyglutamate. Microbiology, 152: 2801–2807.
Nosrati R., Owlia P., Sderi H., Rasooli I., Malbobi M.A. 2014. Phosphate solublization characteristic of efficient nitrogen fixing soil Azotobacter strain. Iranian Journal of Microbiology, 6: 285-295. (In Persian) 
Pandey A., and Palni L.M.S. 1997. Bacillus species: The dominant bacteria of the rhizosphere of established tea. Microbiological Research, 152: 359-365.
Postma J.A., and Lynch J.P. 2011. Root cortical aerenchyma enhances growth of Zea mays L. on soils with suboptimal availability of nitrogen, phosphorus and potassium. Plant Physiology, 156: 1190–1201.
Saiyad S.A., Jhala Y.K., Vyas R.V. 2015. Comparative efficiency of five potash and phosphate solubilizing bacteria and their key enzymes useful for enhancing and improvement of soil fertility. International Journal of Scientific and Research Publications, 5: 1-6.
Saleh-Lakha S., and Glick BR. 2006. Plant growth-promoting bacteria. In: van Elsas J.D., Jansson J.K., Trevors J.T. (Eds) Modern Soil Microbiology. CRC/Thomson Publishing, Boca Raton, FL/UK, pp. 503–520.
Sanger F., Coulson A.R. 1975. A rapid method for determining sequences in DNA by primed synthesis with DNA polymerase. Journal of Molecular Biology, 94 (3): 441–8.
Seneviratne G., Kecskes M.L., and Kennedy I.R. 2008. Biofilmed biofertilisers: Novel inoculants for efficient nutrient use in plants. In: Kennedy I.R., Choudhury A.T., Kecskes M.L., Rose M.T. (Eds.) Efficient nutrient use in rice production in Vietnam achieved using inoculants biofertilisers. Proceedings of a project (SMCN/2002/073) workshop held in Hanoi, Vietnam, 12–13 October 2007. ACIAR Proceeding No. 130, ACIAR, Canberra. pp. 126–130.
Seneviratne G., Weerasekara M.L.M.A.W., Seneviratne K.A.C.N., Zavahir J.S, Kecskes M.L., and Kennedy I.R. 2010. Importance of biofilm formation in plant growth promoting rhizobacterial cction. In: Maheshwari D.K. (Eds) Plant Growth and Health Promoting Bacteria (Microbiology Monographs). Springer-Verlag Berlin Heidelberg.
Shinozaki K., and Yamaguchi-Shinozaki K. 2007. Gene networks involved in drought stress response and tolerance. Journal of Experimental Botany, 58: 221–227.
Shintu P.V., and Jayaram K.M. 2015. Phosphate solubilising bacteria (Bacillus polymyxa) an effective approach to mitigate drought in tomato (Lycopersicon esculentum Mill). Tropical Plant Research. 2:17–22.
Srdjan S., Dragana V., Ivana D., Branislava S., and Milena S.V. 2000. A modified microtiter-plate test for quantification of staphylococcal biofilm formation. Journal of Microbiological Methods, 40:175–179.
Stewart P.S., and Franklin M.J. 2008.  Physiological heterogeneity in biofilms. Nature Reviews Microbiology, 6: 199–210.
Vardharajula S., Ali S.Z., Grover M., Reddy G., and Venkateswaralu B. 2010. Effect of plant growth promoting Pseudomonas spp. on compatible solutes antioxidant status and plant growth of maize under drought stress. Journal of Plant Growth Regulation, 62: 21–30.
Wang C.J., Yang W., Wang C., Gu C., Niu D.D., Liu H.X., Wang Y.P., and Guo J.H. 2012. Induction of drought tolerance in cucumber plants by a consortium of three plant growth-promoting rhizobacterium strains. PLoS One, 7: 525-565.
Yan F., Yu Y., Wang  L., Luo Y., Guo J., and Chai Y. 2016. The comER Gene Plays an Important Role in Biofilm Formation and Sporulation in both Bacillus subtilis and Bacillus cereus. Frontier in Microbiology, 7: 1025-1037.
Zhang C., and Kong F. 2014. Isolation and identification of potassium-solubilizing bacteria from tobacco rhizospheric soil and their effect on tobacco plants. Applied Soil Ecology, 82: 18–25.
Zhang H., Kim M.S., Krishnamachari V., Payton P., Sun Y., Grimson M., Farag M.A., Ryu, C.M. 2007. Rhizobacterial volatile emissions regulate auxin homeostasis and cell expansion in Arabidopsis. Planta, 226: 839–851.

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  • تاریخ دریافت: 16 مرداد 1396
  • تاریخ بازنگری: 26 مرداد 1398
  • تاریخ پذیرش: 28 بهمن 1396

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