جداسازی میکروارگانیسم‌های حل‌کننده فسفات از ریزوسفر گندم و بررسی توان حل-کنندگی آنها در دو منبع فسفات نامحلول

نوع مقاله: مقاله پژوهشی

نویسندگان

1 دانشجوی کارشناسی ارشدگروه علوم خاک دانشکده کشاورزی دانشگاه ارومیه

2 دانشیار گروه علوم خاک، دانشکده کشاورزی دانشگاه ارومیه

3 استادیار گروه علوم خاک، دانشکده کشاورزی دانشگاه ارومیه

چکیده

فسفر یکی از عناصر ضروری برای رشد گیاهان بوده و در اغلب خاک­ها رفتار پیچیده­ای داشته و با اجزا خاک بصورت ترکیبات کم محلول تا نامحلول در می­آید. بهره­برداری از پتانسیل بیولوژیک خاک­ها می­تواند حلالیت و تحرک آن را برای تغذیه مطلوب گیاهان فراهم نماید. این تحقیق به­منظور جداسازی و غربالگری سویه­های باکتریایی (PSB) و قارچی (PSF) حل­کننده فسفات از نظر توان انحلال فسفات­ها در حضور دو منبع تری­کلسیم فسفات (TCP) و خاک فسفات (RP) انجام گردید. تعداد 55 نمونه ریزوسفری از مزارع گندم ارومیه برداشت و جداسازی اولیه و ارزیابی توان حل­کنندگی بصورت کیفی در محیط کشت جامد NBRIP انجام شد. توانایی انحلال کمی سویه­های کارآمد در آزمایشی به­صورت فاکتوریل و در قالب طرح کاملاً تصادفی شامل منبع فسفاته (TCP و RP) و تلقیح میکروبی (شاهد، PSB­45­، PSB15­، PSB30، PSB2، PSB­11، PSB12، PSB20، PSF1، PSF3، PSF4 و PSF7) در هفت زمان انکوباسیون در دمای 28 درجه سلسیوس در انکوباتور (0، 1، 3، 6، 9، 12 و 15 روز ) مورد سنجش قرار گرفت. نتایج نشان داد که منبع فسفاته، سویه­های میکروبی و زمان انکوباسیون تاثیر معنی­داری (P<0.001) بر مقادیر فسفر حل­شده و pH محیط داشت. بیشترین میزان فسفر محلول با 636 میکروگرم بر میلی­لیتر توسط سویه­ی PSF1 از منبع تری­کلسیم فسفات آزاد شد. همچنین سویه­ی PSF1 در روز دوازدهم با 570 میکروگرم بر میلی­لیتر بیشترین توان حلالیت را نسبت به سایر سویه­ها نشان داد. میزان فسفر حل­شده توسط سویه­ها با pH محیط کشت رابطه­ی خطی منفی و معنی­داری نشان داد. میزان pH در نمونه تلقیح شده با سویه­ی PSF1 در منبع تری­کلسیم فسفات بیشترین مقدار کاهش (4 واحد) را نسبت به سایر سویه­ها و تیمار شاهد نشان داد و همچنین بیشترین کاهش pH در روز دوازدهم (90/3pH=) توسط همین سویه مشاهده شد درحالی که میزان pH در تیمار شاهد بدون تغییر باقی ماند.

کلیدواژه‌ها


عنوان مقاله [English]

Isolation of phosphate-solubilizing microorganisms from wheat rhizosphere and evaluation of the their solubilizing potential in presence of two insoluble phosphate sources

نویسندگان [English]

  • Razieh Ebrahimi Karim Abad 1
  • Mir Hasan Rasouli Sadaghiani 2
  • Mohsen Barin 3
چکیده [English]

Phosphorus (P) is one of nutrient elements for plant growth. In most soils P has complex behavior and forms sparingly soluble and insoluble compounds with soil particles. Phosphorus bioavailability can be controlled by soil biological activities for optimum nutrition of plants. This study was carried out in order to isolation of P-solubilizing bacteria (PSB) as well as P-solubilizing fungi (PSF) and their screening in terms of P solubilizing potential in presence of tricalcium phosphate (TCP) and rock phosphate (RP). A total of 55 soil samples were taken from the wheat rhizosphere of Urmia region, and the isolation as well as qualitative solubilizing potential was done in solid NBRIP media. Quantitative P-dissolution ability of isolated strains were assessed in a factorial experiment based on completely randomized design including phosphate sources (RP and TCP) and selected microbial isolates at incubation condition at 28 degrees Celsius. The studied strains included control, PSB45, PSB15, PSB30, PSB2, PSB11, PSB12, PSB20, PSF1, PSF3, PSF4 and PSF7 and sampling were done at seven incubation times (0, 1, 3, 6, 9, 12 and 15 days). The results showed that P solubilization and pH of medium significantly (P <0.001) influenced by insoluble phosphate sources, microbial isolates and incubation time. So that the maximum release of soluble P (636 µg ml-1) was observed by strain PSF1 from TCP source. Furthermore, the PSF1 strain on the day-12 showed the highest solubilized P (570 µg ml-1) compared to other strains. Negative significant correlation was observed between medium pH and dissolved. The pH levels in treatment inoculated with strain PSF1 on TCP (approximately 4 units) showed the highest pH decrement compared to other strains and control media. Highest decline of pH was observed in day-12 (pH = 3.90) by the same strain, while the pH in the control treatments unvaried. 

Abd-Elmonem, E.A., and Amberger, A. (2000). Studies on some factors affecting the solubilization of P from rock phosphate. In: X International Colloquium for the Optimization of Plant Nutrition, eds. Wally, Y. and Shehab, X. National Research Centre (NRC), Cairo, Egypt, pp. 6-7.

Ahmad, N., and Jha, K. K. (1968). Solubilization of rock phosphate by micro-organisms isolated from Bihar soils. The Journal of General and Applied Microbiology, 14(1), 89-95.

Alikhani, H. A., Saleh-Rastin, N., and Antoun, H. (2007). Phosphate solubilization activity of rhizobia native to Iranian soils. In: First International Meeting on Microbial Phosphate Solubilization, Springer Netherlands, pp. 35-41.

Asea, P. E. A., Kucey, R. M. N., and Stewart, J. W. B. (1988). Inorganic phosphate solubilization by two Penicillium species in solution culture and soil. Soil Biology and Biochemistry, 20(4), 459-464.

Cotteni, A. (1980). Methods of Plant Analysis. In: Soil and Plant Testing FAO Soils Bulletin, NO 38/2, pp. 64-100.

Cunningham, J. E., and Kuiack, C. (1992). Production of citric and oxalic acids and solubilization of calcium phosphate by Penicillium bilaiiApplied and Environmental Microbiology, 58(5), 1451-1458.

Dalai, R. C. (1977). Soil organic phosphorus. Advances in Agronomy, 29, 83-117.

Deubel, A., Gransee, A., and Merbach, W. (2000). Transformation of organic rhizodeposits by rhizoplane bacteria and its influence on the availability of tertiary calcium phosphate. Journal of Plant Nutrution and Soil Science, 163, 387-392.

Edi–Premono, M., Moawad,A., and Vleck, P.L.G. (1996). Effect of phosphate solubilizing Pseudmonas putida on the growth of maize and its survival in the rhizosphere. Indonasian Journal of Crop Science, 11, 13–23.

Ghaderi, A., Aliasgharzad, N., Oustan, S. and Olsson, P.A. (2008). Efficiency of three Pseudomonas isolates in releasing phosphate from an artificial variable charge mineral (iron III hydroxide). Soil Environment, 27, 71-76.

Goenadi, D.H., Siswanto, M. and Sugiaro, Y. (2000). Bioactivation of poorly soluble phosphate rocks with a phosphorus-solubilizing fungus. Soil Science Society of America Journal, 64, 927–932.

Guinazu, L.B, Andres, J.A., Mfdel, P., Pistorio, M. and Rosas, S.B. (2010). Response of alfalfa (Medicago sativa L.) to single and mixed inoculation with phosphate-solubilizing bacteria and Sinorhizobium meliloti. Biology and Fertility of Soils, 46, 185–190.

Gupta, N., Sabat, J., Parida, R., and Kerkatta, D. (2007). Solubilization of tricalcium phosphate and rock phosphate by microbes isolated from chromite, iron and manganese mines. Acta Botanica Croatica, 66(2), 197-204.

Jung, I., Park, D. H., and Park, K. (2002). A study of the growth condition and solubilization of phosphate from hydroxyapatite by Pantoea agglomerans. Biotechnology and Bioprocess Engineering, 7(4), 201-205.

Kang, S. C., Pandey, P., Khillon, R., and Maheshwari, D. K. (2008). Process of rock phosphate solubilization by Aspergillus sp PS 104 in soil amended medium. Journal of Environmental Biology, 29(5), 743-746.

Kannapiran, E., and Ramkuma, V. S. (2011). Isolation of phosphate solubilizing bacteria from sediments of Thondi coast, Palk Strait, Southeast Coast India. Annals of Biological Research, 2, 157-163.

Keneni, A., Assefa, F., and Prabu, P. C. (2010). Isolation of phosphate solubilizing bacteria from the rhizosphere of faba bean of Ethiopia and their abilities on solubilizing insoluble phosphates. Journal of Agricultural Science and Technology, 12, 79-89.

Kucey, R. M. N. (1983). Phosphate-solubilizing bacteria and fungi in various cultivated and virgin Alberta soils. Canadian Journal of Soil Science, 63(4), 671-678.

Malakooti, M. J. (1998). Sustainable agriculture and increase performance by optimizing the use of fertilizers in Iran. Publish Agricultural Training. Karaj, Iran, 460p. (in Persian).

Malakooti, M. J. and Riazi Hamadani, S.A. (1990). Fertilizers and soil fertility. First Edition, The University of Tehran Press, 808p. (in Persian).

Mehnaz, S., and Lazarovits, G. (2006). Inoculation effects of Pseudomonas putida, Gluconacetobacter azotocaptans, and Azospirillum lipoferum on corn plant growth under greenhouse conditions. Microbial Ecology, 51(3), 326-335.

Meunchang, S., Thongra-Ar, P., Sanoh, S., Kaewsuralikhit, S., and Ando, S. (2006). Development of rhizobacteria as a biofertilizer for rice production. In: International workshop on Sustained Management of the Soil-Rhizosphere System for Efficient Crop Production and Fertilizer Use, pp. 16-20.

Mittal, V., Singh, O., Nayyar, H., Kaur, J., and Tewari, R. (2008). Stimulatory effect of phosphate-solubilizing fungal strains (Aspergillus awamori and Penicillium citrinum) on the yield of chickpea (Cicer arietinum L. cv. GPF2). Soil Biology and Biochemistry, 40(3), 718-727.

Miyasaka, S. C., and Habte, M. (2001). Plant mechanisms and mycorrhizal symbioses to increase phosphorus uptake efficiency. Communications in Soil Science and Plant Analysis, 32(7-8), 1101-1147.

Molla, A. H., Shamsuddin, Z. H., Halimi, M. S., Morziah, M., and Puteh, A. B. (2001). Potential for enhancement of root growth and nodulation of soybean co-inoculated with Azospirillum and Bradyrhizobium in laboratory systems. Soil Biology and Biochemistry, 33(4), 457-463.

Nahas, E. (1996). Factors determining rock phosphate solubilization by microorganism isolated from soil. World Journal Microbiology Biotechnology, 12(6), 18-23.

Narsian, V., and Patel, H. H. (2000). Aspergillus aculeatus as a rock phosphate solubilizer. Soil Biology and Biochemistry, 32(4), 559-565.

Nautiyal, C. S. (1999). An efficient microbiological growth medium for screening phosphate solubilizing microorganisms. FEMS Microbiology Letters, 170(1), 265-270.

Oliveira, C. A., Alves, V. M. C., Marriel, I. E., Gomes, E. A., Scotti, M. R., Carneiro, N. P., Guimaraes, C., Schaffert, R.E. and Sa, N. M. H. (2009). Phosphate solubilizing microorganisms isolated from rhizosphere of maize cultivated in an oxisol of the Brazilian Cerrado Biome. Soil Biology and Biochemistry, 41(9), 1782-1787.

Rajan SSS, Watkinson JH and Sinclair AG 1996. Phosphate rocks for direct application to soil. Advance in Agronomy, 57, 77-14.

Rajan, S. S. S., Watkinson, J. H., and Sinclair, A. G. (1996). Phosphate rocks for direct application to soils. Advances in Agronomy, 57, 77-159.

Rasipoor, L. and, Aliasgharzad, N. (2005). Interaction of phosphate solubilizing bacteria and Bradyrhizobium japonicum on yield and nutrient uptake by Soybean. Agriculture Science, 15, 141-156, (In Persian).

Reddy, M. S., Kumar, S., Babita, K., and Reddy, M. S. (2002). Biosolubilization of poorly soluble rock phosphates by Aspergillus tubingensis and Aspergillus nigerBioresource Technology, 84(2), 187-189.

Rejali, f., Asadi, H., Khavazi, k., Asgharzadeh, A., and Afshari, M. (2010). The status of biological phosphate fertilizers and the necessity of  its development in Iranian agricultural. The 1st Iranian Fertilizer Challenges Congress: Half a Century of the Fertilizer Consumption. 1-3 March, Tehran, Iran, (in Persian).

Reyes I, Bernier L, Simard R, Tanguay P. H and Antoun H. 1999.  Characteristics of phosphate solubilization by an isolate of a tropical Penicillium rugulosum and two UV-induced mutants. FEMS Microbiology Ecology, 28, 291–295.

Rodriguez, H., and Frage, R. (1999). Phosphate solubilizing bacteria and their role in plant growth promotion. Biotechnology Advances, 17, 319-339.

Shankarrao, O. (2012). Isolation and characterization of phosphate solubilizing bacteria from rhizospheric Soil Samples. International Interdisciplinary Research Journal, 2(4), 28-29.

Sharma, S., Kumar, V., and Tripathi, R. B. (2011). Isolation of phosphate solubilizing microorganism (PSMs) from soil. Journal Microbial Biotechnology Research, 1(2), 90-95.

Sharma, K. (2005). Isolation, Purification and Identification of Bacteria. In: Manual of Microbiology, Ane Books Publication, New Delhi, 41p.

Turan, M., Ataoglu, N., and Sahιn, F. (2006). Evaluation of the capacity of phosphate solubilizing bacteria and fungi on different forms of phosphorus in liquid culture. Journal of Sustainable Agriculture, 28(3), 99-108.

Vance, C. P., Uhde‐Stone, C., and Allan, D. L. (2003). Phosphorus acquisition and use: critical adaptations by plants for securing a nonrenewable resource. New Phytologist, 157(3), 423-447.

Vassilev, N., Vassileva, M., Fenice, M., and Federici, F. (2001). Immobilized cell technology applied in solubilization of insoluble inorganic (rock) phosphates and P plant acquisition. Bioresource Technology, 79(3), 263-271.

Vazquez, P., Holguin, G., Puente, M. E., Lopez-Cortes, A., and Bashan, Y. (2000). Phosphate-solubilizing microorganisms associated with the rhizosphere of mangroves in a semiarid coastal lagoon. Biology and Fertility of Soils, 30(5-6), 460-468.

Vyas, P., and Gulati, A. (2009). Organic acid production in vitro and plant growth promotion in maize under controlled environment by phosphate-solubilizing fluorescent PseudomonasBMC Microbiology, 9(1), 1.

Venkateswarlu, B., Rao, A. V., and Raina, P. (1984). Evaluation of phosphorus solubilisation by microorganisms isolated from Aridisols. Journal of the Indian Society of Soil Science, 32(2), 273-277.

Whitelaw, M. A. (1999). Growth promotion of plants inoculated with phosphate-solubilizing fungi. Advances in Agronomy, 69, 99-151.

Yu, X., Liu, X., Zhu, T. H., Liu, G. H., and Mao, C. (2011). Isolation and characterization of phosphate-solubilizing bacteria from walnut and their effect on growth and phosphorus mobilization. Biology and Fertility of Soils, 47(4), 437-446.