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Isolation and characterization of phosphate-solubilizing bacteria from walnut and their effect on growth and phosphorus mobilization

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Abstract

The objectives of this work were to isolate and characterize walnut phosphate-solubilizing bacteria (PSB) and to evaluate the effect of inoculation with the selected PSB stains to walnut seedlings fertilized with or without insoluble phosphate. Thirty-four PSB strains were isolated and identified under the genera Pseudomonas, Stenotrophomonas, Bacillus, Cupriavidus, Agrobacterium, Acinetobacter, Arthrobacter, Pantoea, and Rhodococcus through a comparison of the 16S ribosomal DNA sequences. All isolated PSB strains could solubilize tricalcium phosphate (TCP) in solid and liquid media. Phosphate-solubilizing activity of these strains was associated with a drop in the pH of medium. A significantly negative linear correlation was found between culture pH and phosphorus (P) solubilized from inorganic phosphate. Three isolates Pseudomonas chlororaphis (W24), Bacillus cereus (W9), and Pseudomonas fluorescens (W12) were selected for shade house assays because of their higher phosphate-solubilizing abilities. Under shade house conditions, application of W24 or W12 remarkably improved plant height, shoot and root dry weight, and P and nitrogen (N) uptake of walnut seedlings. These increases were higher on combined inoculation of PSB with TCP addition. The most pronounced beneficial effect on growth of walnut plants was observed in the co-inoculation of the three PSB strains with TCP addition. In comparison, the isolate of W9 failed to increase available soil P, nutrient levels in plants, or to promote plant growth, suggesting that more insoluble phosphate compounds than tricalcium phosphate should be used as substrates to assess the phosphate-solubilizing ability of PSB under greenhouse conditions. The present results indicated that strains P. chlororaphis or P. fluorescens could be considered for the formulation of new inoculants of walnut, even of more woody plants.

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References

  • Afzal A, Bano A (2008) Rhizobium and phosphate solubilizing bacteria improve the yield and phosphate uptake in wheat (Triticum aestivum). Int J Agric boil 10:85–88

    CAS  Google Scholar 

  • Akhtar MS, Siddiqui ZA (2009) Effects of phosphate solubilizing microorganisms and Rhizobium sp. on the growth, nodulation, yield and root-rot disease complex of chickpea under field condition. Afr J biotechnol 8:3489–3496

    Google Scholar 

  • Alagawadi AR, Gaur AC (1992) Inoculation of Azospirillum brasilense and phosphate-solubilizing bacteria on yield of sorghum [Sorghum bicolor (L.) Moench] in dry land. Trop Agric 69:347–350

    Google Scholar 

  • Alam MM, Ladha JK (2004) Optimizing phosphorus fertilization in an intensive vegetable–rice cropping system. Biol Fertil Soils 40:277–283

    Article  Google Scholar 

  • Aslantas R, Cakmakci R, Sahin F (2007) Effect of plant growth promoting rhizobacteria on young apple tree growth and fruit yield under orchard conditions. Sci Hortic 111:371–377

    Article  Google Scholar 

  • Basak BB, Biswas DR (2010) Co-inoculation of potassium solubilizing and nitrogen fixing bacteria on solubilization of waste mica and their effect on growth promotion and nutrient acquisition by a forage crop. Biol Fertil Soils 46:641–648

    Article  Google Scholar 

  • Chanway CP (1997) Inoculation of tree roots with plant growth promoting soil bacteria: an emerging technology for reforestation. For Sci 1:99–112

    Google Scholar 

  • Chung H, Park M, Madhaiyan M, Seshadri S, Song J, Cho H, Sa T (2005) Isolation and characterization of phosphate solubilizing bacteria from the rhizosphere of crop plants of Korea. Soil Boil Biochem 37:1970–1974

    Article  CAS  Google Scholar 

  • Collavino MM, Sansberro PA, Mroginski LA, Mario Aguilar O (2010) Comparison of in vitro solubilization activity of diverse phosphate-solubilizing bacteria native to acid soil and their ability to promote Phaseolus vulgaris growth. Biol Fertil Soils 46:727–738

    Article  Google Scholar 

  • Colwell JD (1965) An automatic procedure for the determination of phosphorus in sodium hydrogen carbonate extracts of soils. Chem Ind 22:893–895

    Google Scholar 

  • Das AC (1989) Utilization of insoluble phosphates by soil fungi. J Indian Soc Soil Sci 58:1208–1211

    Google Scholar 

  • De Freitas JR, Banerjee MR, Germida JJ (1997) Phosphate-solubilizing rhizobacteria enhance the growth and yield but not phosphorus uptake of canola (Brassica napus L.). Biol Fertil Soils 24:358–364

    Article  Google Scholar 

  • Delvasto P, Valverde A, Ballester A, Igual JM, Muñoz JA, Gonzáles F, Blázquez ML, García C (2006) Characterization of brushite as a re-crystallization product formed during bacterial solubilization of hydroxyapatite in batch cultures. Soil Biol Biochem 38:2645–2654

    Article  CAS  Google Scholar 

  • Fernandez LA, Zalba P, Gomez MA, Sagarday MA (2007) Phosphate-solubilization activity of bacterial strains in soil and their effect on soybean growth under greenhouse conditions. Biol Fertil Soils 43:805–809

    Article  CAS  Google Scholar 

  • Goldstein AH (1995) Recent progress in understanding the molecular genetics and biochemistry of calcium phosphate solubilization by Gram negative bacteria. Biol Agric Hort 12:185–193

    Google Scholar 

  • Guiñazú LB, Andrés JA, MFDel P, Pistorio M, Rosas SB (2010) Response of alfalfa (Medicago sativa L.) to single and mixed inoculation with phosphate-solubilizing bacteria and Sinorhizobium meliloti. Biol Fertil Soils 46:185–190

    Article  Google Scholar 

  • Gulati A, Sharma N, Vyas P, Sood S, Rahi P, Pathania V, Prasad R (2010) Organic acid production and plant growth promotion as a function of phosphate solubilization by Acinetobacter rhizosphaerae strain BIHB 723 isolated from the cold deserts of the trans-Himalayas. Arch Microbiol 192:975–983

    Article  PubMed  CAS  Google Scholar 

  • Gull M, Hafeez FY, Saleem M, Malik A (2004) Phosphate uptake and growth promotion of chickpea by co-inoculation of mineral phosphate solubilizing bacteria and a mixed rhizobial culture. Aust J Exp Agric 44:623–628

    Article  CAS  Google Scholar 

  • Gyaneshwar PG, Naresh Kumar G, Parekh LJ, Poole PS (2002) Role of soil microorganisms in improving P nutrition of plants. Plant Soil 245:83–93

    Article  CAS  Google Scholar 

  • Halder AK, Mishra AK, Bhattacharya P, Chakrabarthy PK (1990) Solubilization of rock phosphate by Rhizobium and Bradyrhizobium. J Gen Appl Microbiol 36:81–92

    Article  CAS  Google Scholar 

  • Hameeda B, Harini G, Rupela OP, Wani SP, Reddy G (2008) Growth promotion of maize by phosphate-solubilizing bacteria isolated from composts and macrofauna. Microbiol Res 163:234–242

    Article  PubMed  CAS  Google Scholar 

  • Hwangbo H, Park RD, Kim YW, Rim YS, Park KH, Kim TH, Suh JS, Kim KY (2003) 2-Ketogluconic production and phosphate solubilization by Enterobacter intermedium. Curr Microbiol 47:87–92

    Article  PubMed  CAS  Google Scholar 

  • Illmer P, Schinner F (1995) Solubilization of inorganic calcium phosphates—solubilization mechanisms. Soil Biol Biochem 27:257–263

    Article  CAS  Google Scholar 

  • Jisha MS, Alagawadi AR (1996) Nutrient uptake and yield of sorghum [(Sorghum bicolor (L.) Moench)] inoculated with phosphate solubilizing bacteria and cellulolytic fungus in a cotton stalk amended vertisol. Microbiol Res 151:213–217

    CAS  Google Scholar 

  • Johri JK, Surange S, Nautiyal CS (1999) Occurrence of salt, pH, and temperature-tolerant, phosphate-solubilizing bacteria in alkaline soils. Curr Microbiol 39:89–93

    Article  PubMed  CAS  Google Scholar 

  • Jones JB, Wolf B, Mills HA (1991) Plant analysis handbook. Micro-Macro Publishing, Georgia

    Google Scholar 

  • Kim KY, Jordan D, Mcdonald GA (1998) Enterobacter agglomerans, phosphate solubilizing bacteria, and microbial activity in soil: effect of carbon source. Soil Biol Biochem 30:995–1003

    Article  CAS  Google Scholar 

  • Kpomblekou K, Tabatabai MA (1994) Effect of organic acids on release of phosphorus from phosphate rocks. Soil Sci 158:442–453

    Article  Google Scholar 

  • Mukherjee PK, Rai RK (2000) Effect of vesicular arbuscular mycorrhizae and phosphate-solubilizing bacteria on growth, yield and phosphorus uptake by wheat (Triticum aestivum) and chickpea (Cicer arietinum). Indian J agronomy 45:602–607

    Google Scholar 

  • Nautiyal CS (1999) An efficient microbiological growth medium for screening phosphate solubilizing microorganisms. FEMS Microbiol Lett 182:265–270

    Article  Google Scholar 

  • Peng JB (1989) Pomiculture. Chinese Agriculture Press, Beijing

    Google Scholar 

  • Pikovskaya RI (1948) Mobilization of phosphorus in soil in connection with vital activity of some microbial species. Microbiologiya 17:362–370

    CAS  Google Scholar 

  • Piper CS (1967) Soil and plant analysis. Asia Publishing House, Bombay

    Google Scholar 

  • Puente ME, Li CY, Bashan Y (2004) Microbial populations and activities in the rhizoplane of rock-weathering desert plants. II. Growth promotion of cactus seedlings. Plant Biol 6:643–650

    Article  PubMed  CAS  Google Scholar 

  • Rashid M, Khalil S, Ayub N, Alam S, Latif F (2004) Organic acids production and phosphate solubilization by phosphate solubilizing microorganisms (PSM) under in vitro conditions. Pak J Biol Sci 7:187–196

    Article  Google Scholar 

  • Reddy MS, Kumar S, Babita K (2002) Biosolubilization of poorly soluble rock phosphates by Aspergillus tubingensis and Aspergillus niger. Bioresour Technol 84:187–189

    Article  PubMed  CAS  Google Scholar 

  • Richardson AE (2001) Prospects for using soil microorganisms to improve the acquisition of phosphorus by plants. Aust J Plant Physiol 28:897–906

    Google Scholar 

  • Rodriguez H, Fraga R (1999) Phosphate solubilizing bacteria and their role in plant growth promotion. Biotechnol Adv 17:319–339

    Article  PubMed  CAS  Google Scholar 

  • Rudresh DL, Shivaprakash MK, Prasad RD (2005) Effect of combined application of Rhizobium, phosphate solubilizing bacterium and Trichoderma spp. on growth, nutrient uptake and yield of chickpea (Cicer aritenium L.). Appl Soil Ecol 28:139–146

    Article  Google Scholar 

  • Sahin F, Cakmakci R, Kantar F (2004) Sugar beet and barely yields in relation to inoculation with N2-fixing and phosphate solubilizing bacteria. Plant Soil 265:123–129

    Article  CAS  Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Press, New York

    Google Scholar 

  • Selvaraj P, Madhaiyan M, Sa T (2008) Isolation and identification of phosphate solubilizing bacteria from Chinese cabbage and their effect on growth and phosphorus utilization of plants. J Microbiol Biotechnol 18:773–777

    Google Scholar 

  • Singh S, Kapoor KK (1999) Inoculation with phosphate-solubilizing microorganisms and a vesicular–arbuscular mycorrhizal fungus improves dry matter yield and nutrient uptake by wheat grown in a sandy soil. Biol Fertil Soils 28:139–144

    Article  CAS  Google Scholar 

  • Suslow TV, Schroth MN, Isaka M (1982) Application of a rapid method for Gram differentiation of plant pathogentic and saprophytic bacteria without staining. Phytopathology 72:917–918

    Article  Google Scholar 

  • Tao GC, Tian SJ, Cai MY, Xie GH (2008) Phosphate-solubilizing and mineralizing abilities of bacteria isolated from soils. Pedosphere 18:515–523

    Article  CAS  Google Scholar 

  • Taurian T, Anzuay MS, Angelini JG, Tonelli ML, Luduena L, Pena D, Inanez F, Fabra A (2010) Phosphate-solubilizing peanut associated bacteria: screening for plant growth-promoting activities. Plant Soil 329:421–431

    Article  CAS  Google Scholar 

  • Vassilev N, Vassileva M (2003) Biotechnological solubilization of rock phosphate on media containing agroindustrial wastes. Appl Microbiol Biotechnol 61:435–440

    PubMed  CAS  Google Scholar 

  • Vonderwell JD, Enebak SA (2000) Different effects of rhizobacterial strain and dose on the ectomycorrhizal colonization of loblolly pine seedlings. For Sci 46:411–437

    Google Scholar 

  • Wani PA, Khan MS, Zaidi A (2007) Co-inoculation of nitrogen fixing and phosphate solubilizing bacteria to promote growth, yield and nutrient uptake in chickpea. Acta Agron Hung 55:315–323

    Article  CAS  Google Scholar 

  • Watanabe FS, Olsen SR (1965) Test of an ascorbic acid method for determining phosphorus in water and NaHCO3 extracts from soils. Soil Sci Soc Am J 29:677–678

    Article  CAS  Google Scholar 

  • Zhang PC, Shao GF, Zha G, Le Master DC, Parker GR, Dunning JB, Li QL (2000) China’s forest policy for 21st century. Science 288:2135–2136

    Article  PubMed  CAS  Google Scholar 

  • Zinniel DK, Lambrech P, Harris NB, Feng Z, Kuczmarski D, Higley P, Ishimaru CA (2002) Isolation and characterization of endophytic colonizing bacteria from agronomic crops and prairie plants. Appl Environ Microbiol 68:2198–2208

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

This study was supported by State Natural Science & Technology Resources’ Share Base of P. R. China (2005DKA21207-13). We thank Wanbo Wu, Huabai Han, Sichuan Academy of Forestry for providing experimental walnut seedlings.

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Authors and Affiliations

  1. College of Forestry, Sichuan Agricultural University, Ya’an, 625014, Sichuan, China

    Xuan Yu, Tian Hui Zhu, Guang Hai Liu & Cui Mao

  2. College of Enology, Northwest A&F University, Yangling, 712100, Shaanxi, China

    Xu Liu

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  1. Xuan Yu
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Correspondence to Tian Hui Zhu.

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Yu, X., Liu, X., Zhu, T.H. et al. Isolation and characterization of phosphate-solubilizing bacteria from walnut and their effect on growth and phosphorus mobilization. Biol Fertil Soils 47, 437–446 (2011). https://doi.org/10.1007/s00374-011-0548-2

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  • DOI: https://doi.org/10.1007/s00374-011-0548-2

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