Selection and evaluation of phosphate-solubilizing bacteria from grapevine rhizospheres for use as biofertilizers

  • Min Liu Northwest A & F University, College of Enology, Yangling 712100, Shaanxi
  • Xu Liu Northwest A & F University, College of Enology, Yangling 712100, Shaanxi
  • Bao-Sen Cheng Northwest A & F University, College of Enology, Yangling 712100, Shaanxi
  • Xue-Lei Ma Northwest A & F University, College of Enology, Yangling 712100, Shaanxi
  • Xiao-Tong Lyu Northwest A & F University, College of Enology, Yangling 712100, Shaanxi
  • Xian-Fang Zhao Northwest A & F University, College of Enology, Yangling 712100, Shaanxi
  • Yan-Lun Ju Northwest A & F University, College of Enology, Yangling 712100, Shaanxi
  • Zhuo Min Northwest A & F University, College of Enology, Yangling 712100, Shaanxi
  • Yu-Lin Fang Northwest A & F University, College of Enology, Yangling 712100, Shaanxi
Keywords: promote plant growth, inorganic phosphate, stress conditions, saline-alkaline soil, Vitis vinifera L. cv. Cabernet Sauvignon


Phosphate-solubilizing bacteria (PSB) have the ability to solubilize insoluble phosphorus (P) and release soluble P. Extensive research has been performed with respect to PSB isolation from the rhizospheres of various plants, but little is known about the prevalence of PSB in the grapevine rhizosphere. In this study, we aimed to isolate and identify PSB from the grapevine rhizosphere in five vineyards of Northwest China, to characterize their plant-growth-promoting (PGP) traits, evaluate the effect of stress on their phosphate-solubilizing activity (PSA), and test their ability to stimulate the growth of Vitis vinifera L. cv. Cabernet Sauvignon. From the vineyard soils, 66 PSB isolates were screened, and 10 strains with high PSA were identified by 16S rRNA sequencing. Sequence analysis revealed that these 10 strains belonged to 4 genera and 5 species: Bacillus aryabhattai, B. megaterium, Klebsiella variicola, Stenotrophomonas rhizophila, and Enterobacter aerogenes. The selected PSB strains JY17 (B. aryabhattai) and JY22 (B. aryabhattai) were positive for multiple PGP traits, including nitrogen fixation and production of indole acetic acid (IAA), siderophores, 1-aminocyclopropane-1-carboxylate (ACC) deaminase, chitinase, and protease. JY17 and JY22 showed strong PSA under stress conditions of high pH, high salt, and high temperature. Therefore, these two isolates can be used as biofertilizers in saline-alkaline soils. The inoculation with PSB significantly facilitated the growth of V. vinifera cv. Cabernet Sauvignon under greenhouse conditions. Use of these PSB as biofertilizers will increase the available P content in soils, minimize P-fertilizer application, reduce environmental pollution, and promote sustainable agriculture.


Download data is not yet available.


Acevedo E, Galindo-Castañeda T, Prada F, Navia M, Romero HM, 2014. Phosphate-solubilizing microorganisms associated with the rhizosphere of oil palm (Elaeis guineensis Jacq.) in Colombia. Appl Soil Ecol 80(8): 26-33. https:/

Adhikary H, Sanghavi PB, Macwan SR, Archana G, Kumar GN, 2014. Artificial citrate operon confers mineral phosphate solubilization ability to diverse Fluorescent pseudomonads. Plos One 9 (9): 1-12. https:/

Anzuay MS, Frola O, Angelini JG, Ludueña LM, Ibañez F, Fabra A, Taurian T, 2015. Effect of pesticides application on peanut (Arachis hypogaea L.) associated phosphate solubilizing soil bacteria. Appl Soil Ecol 95: 31-37. https:/

Arcand MM, Schneider KD, 2006. Plant- and microbial-based mechanisms to improve the agronomic effectiveness of phosphate rock: A review. Anais Da Academia Brasileira De Ciências 78 (4): 791-807. https:/

Babu-Khan S, Yeo TC, Martin WL, Duron MR, Rogers RD, Goldstein AH, 1995. Cloning of a mineral phosphate-solubilizing gene from Pseudomonas Cepacia. Appl Environ Microbiol 61 (3): 972-978.

Bakker AW, Schippers B, 1987. Microbial cyanide production in the rhizosphere in relation to potato yield reduction and Pseudomonas spp-mediated plant growth-stimulation. Soil Biol Biochem 19 (4): 451-457. https:/

Banerjee S, Palit R, Sengupta C, Standing D, 2010. Stress induced phosphate solubilization by Arthrobacter sp. and Bacillus sp. isolated from tomato rhizoshpere. Aust J Crop Sci 4 (6): 378-383.

Bao S, 2000. Soil agricultural chemistry analysis. Agriculture Press, Beijing, China.

Beneduzi A, Moreira F, Costa PB, Vargas LK, Lisboa BB, Favreto R, Baldani JI, Passaglia LMP, 2013. Diversity and plant growth promoting evaluation abilities of bacteria isolated from sugarcane cultivated in the South of Brazil. Appl Soil Ecol 63: 94-104. https:/

Buch A, Archana G, Naresh Kumar G, 2008. Metabolic channeling of glucose towards gluconate in phosphate-solubilizing Pseudomonas aeruginosa P4 under phosphorus deficiency. Res Microbiol 159 (9-10): 635-642. https:/

Castagno LN, Estrella MJ, Sannazzaro AI, Grassano AE, Ruiz OA, 2011. Phosphate-solubilization mechanism and in vitro plant growth promotion activity mediated by Pantoea eucalypti isolated from Lotus tenuis rhizosphere in the Salado River Basin (Argentina). J Appl Microbiol 110 (5): 1151-1165. https:/

Chen YP, Rekha PD, Arun AB, Shen FT, Lai WA, Young CC, 2006. Phosphate solubilizing bacteria from subtropical soil and their tricalcium phosphate solubilizing abilities. Appl Soil Ecol 34 (1): 33-41. https:/

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 Biol Biochem 37 (10): 1970-1974. https:/

Dey R, Pal KK, Bhatt DM, Chauhan SM, 2004. Growth promotion and yield enhancement of peanut (Arachis hypogaea L.) by application of plant growth-promoting rhizobacteria. Microbiol Res 159 (4): 371-394. https:/

Fernández LA, Zalba P, Gómez MA, Sagardoy MA, 2007. Phosphate-solubilization activity of bacterial strains in soil and their effect on soybean growth under greenhouse conditions. Biol Fertil Soils 43 (6): 805-809. https:/

Finch-Savage WE, Leubner-Metzger G, 2006. Seed dormancy and the control of germination. New Phytologist 171 (3): 501-523. https:/

Freitas JRD, 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 (24): 358-364. https:/

Gianinetti A, Laarhoven LJJ, Persijn ST, Harren FJM, Petruzzelli L, 2007. Ethylene production is associated with germination but not seed dormancy in red rice. Ann Bot 99(4): 735-745. https:/

Glickmann E, Dessaux Y, 1995. A critical examination of the specificity of the Salkowski reagent for indolic compounds produced by phytopathogenic bacteria. Appl Environ Microbiol 61 (2): 793-796.

Goldstein AH, 1995. Recent progress in understanding the molecular genetics and biochemistry of calcium phosphate solubilization by gram negative bacteria. Biol Agr Hortic 12 (2): 185-193. https:/

Gupta M, Kiran S, Gulati A, Singh B, Tewari R, 2012. Isolation and identification of phosphate solubilizing bacteria able to enhance the growth and aloin-a biosynthesis of Aloe barbadensis Miller. Microbiol Res 167 (6): 358-363. https:/

Halder AK, Mishra AK, Bhattacharyya P, Chakrabartty PK, 1990. Solubilization of rock phosphate by Rhizobium and Bradyrhizobium. J Gen Appl Microbiol 36 (2): 81-92. https:/

Illmer P, Schinner F, 1992. Solubilization of inorganic phosphates by microorganisms isolated from forest soils. Soil Biol Biochem 24 (4): 389-395. https:/

Illmer P, Schinner F, 1995. Solubilization of inorganic calcium phosphates—Solubilization mechanisms. Soil Biol Biochem 27 (3): 257-263. https:/

Johri JK, Surange S, Nautiyal CS, 1999. Occurrence of salt, pH, and temperature-tolerant, phosphate-solubilizing bacteria in alkaline soils. Curr Microbiol 39 (2): 89-93. https:/

Karagöz K, Ateş F, Karagöz H, Kotan R, Çakmakçı R, 2012. Characterization of plant growth-promoting traits of bacteria isolated from the rhizosphere of grapevine grown in alkaline and acidic soils. Eur J Soil Biol 50: 144-150. https:/

Kim KY, Jordan D, Mcdonald GA, 1998. Enterobacter agglomerans, phosphate solubilizing bacteria, and microbial activity in soil: Effect of carbon sources. Soil Biol Biochem 30 (8-9): 995-1003. https:/

Krishnaraj PU, Goldstein AH, 2001. Cloning of a Serratia marcescens DNA fragment that induces quinoprotein glucose dehydrogenase-mediated gluconic acid production in Escherichia coli in the presence of stationary phase Serratia marcescens. FEMS Microbiol Lett 205 (2): 215-220. https:/

Kucera B, Cohn MA, Leubner-Metzger G, 2005. Plant hormone interactions during seed dormancy release and germination. Seed Sci Res 15 (4): 281-307. https:/

Kumar KV, Srivastava S, Singh N, Behl HM, 2009. Role of metal resistant plant growth promoting bacteria in ameliorating fly ash to the growth of Brassica juncea. J Hazard Mater 170 (1): 51-57. https:/

Lin L, Li Z, Hu C, Zhang X, Chang S, Yang L, Li Y, An Q, 2012. Plant growth-promoting nitrogen-fixing enterobacteria are in association with sugarcane plants growing in Guangxi, China. Microb Environ 27 (4): 391-398. https:/

Lugtenberg B, Kamilova F, 2009. Plant-growth-promoting rhizobacteria. Annu Rev Microbiol 63 (63): 541-556. https:/

Mamta, Rahi P, Pathania V, Gulati A, Singh B, Bhanwra RK, Tewari R, 2010. Stimulatory effect of phosphate-solubilizing bacteria on plant growth, Stevioside and Rebaudioside-a contents of Stevia rebaudiana Bertoni. Appl Soil Ecol 46 (2): 222-229. https:/

Marasco R, Rolli E, Fusi M, Cherif A, Abou-Hadid A, El-Bahairy U, Borin S, Sorlini C, Daffonchio D, 2013. Plant growth promotion potential is equally represented in diverse grapevine root-associated bacterial communities from different biopedoclimatic environments. Biomed Res Int 2013 (3): 247-261. https:/

Mayak S, Tirosh T, Glick BR, 2004. Plant growth-promoting bacteria confer resistance in tomato plants to salt stress. Plant Physiol Biochem 42 (6): 565-572. https:/

Nadeem SM, Zahir ZA, Naveed M, Arshad M, 2007. Preliminary investigations on inducing salt tolerance in maize through inoculation with rhizobacteria containing ACC deaminase activity. Can J Microbiol 53 (10): 1141-1149. https:/

Nautiyal CS, 1999. An efficient microbiological growth medium for screening phosphate solubilizing microorganisms. FEMS Microbiol Lett 170 (1): 265-270. https:/

Park JH, Bolan N, Megharaj M, Naidu R, 2011. Isolation of phosphate solubilizing bacteria and their potential for lead immobilization in soil. J Hazard Mater 185 (2-3): 829-836. https:/

Penrose DM, Glick BR, 2003. Methods for isolating and characterizing ACC deaminase-containing plant growth-promoting rhizobacteria. Physiol Plantarum 118: 10-15. https:/

Principe A, Alvarez F, Castro MG, Zachi L, Fischer SE, Mori GB, Jofre E, 2007. Biocontrol and Pgpr features in native strains isolated from saline soils of Argentina. Curr Microbiol 55 (4): 314-322. https:/

Rashid M, Khalil S, Ayub N, Alam S, 2004. Organic acids production and phosphate solubilization by phosphate solubilizing microorganisms (Psm) under in vitro conditions. Pak J Biol Sci 7 (2): 187-196. https:/

Rojas-Avelizapa L, Cruz-Camarillo R, Guerrero M, Rodriguez-Vazquez R, Ibarra J, 1999. Selection and characterization of a proteo-chitinolytic strain of Bacillus thuringiensis, able to grow in shrimp waste media. World J Microb Biot 15: 299-308. https:/

Sagoe CI, Ando T, Kouno K, Nagaoka T, 1998. Relative importance of protons and solution calcium concentration in phosphate rock dissolution by organic acids. Soil Sci Plant Nutr 44 (4): 617-625. https:/

Saravanakumar D, Samiyappan R, 2007. Acc deaminase from Pseudomonas fluorescens mediated saline resistance in groundnut (Arachis hypogea) plants. J Appl Microbiol 102 (5): 1283-1292. https:/

Schwyn B, Neilands JB, 1987. Universal chemical assay for the detection and determination of siderophores. Anal Biochem 160 (1): 47-56. https:/

Shahzad SM, Arif MS, Riaz M, Iqbal Z, Ashraf M, 2013. Pgpr with varied ACC-deaminase activity induced different growth and yield response in maize (Zea mays L.) under fertilized conditions. Eur J Soil Biol 57: 27-34. https:/

Sheik CS, Beasley WH, Elshahed MS, Zhou X, Luo Y, Krumholz LR, 2011. Effect of warming and drought on grassland microbial communities. Isme Journal 5 (10): 1692-1700. https:/

Srividya S, Soumya S, Pooja K, 2009. Influence of environmental factors and salinity on phosphate solubilization by a newly isolated Aspergillus niger F7 from agricultural soil. Afr J Biotechnol 8 (9): 1864-1870.

Sun J, Zhang R, Wang H, Fan W, Luo L, Han J, Ma Z, 2014. Stpatial variation of soil salt content and pH of salinized soil in Dingbian County, Shaanxi Province. Acta Agriculturae Boreali-Occidentalis Sinica 23 (9): 114-119.

Watanabe FS, Olsen SR, 1965. Test of an ascorbic acid method for determining P in water and NaHCO3 extracts from soil. Soil Sci Soc Am Proc 29: 677-678. https:/

Weisburg WG, Barns SM, Pelletier DA, Lane DJ, 1991. 16s ribosomal DNA amplification for phylogenetic study. J Bacteriol 173 (2): 697-703. https:/

Wu Z, Peng Y, Guo L, Li C, 2014. Root colonization of encapsulated Klebsiella oxytoca Rs-5 on cotton plants and its promoting growth performance under salinity stress. Eur J Soil Biol 60: 81-87. https:/

Yadav K, Kumar C, Archana G, Kumar GN, 2014. Artificial citrate operon and Vitreoscilla hemoglobin gene enhanced mineral phosphate solubilizing ability of Enterobacter hormaechei Dhrss. Appl Microbiol Biotechnol 98 (19): 8327-8336. https:/

Yang H, Li Q, 2008. Studies on path for transitin

How to Cite
Liu, M., Liu, X., Cheng, B.-S., Ma, X.-L., Lyu, X.-T., Zhao, X.-F., Ju, Y.-L., Min, Z., & Fang, Y.-L. (2017). Selection and evaluation of phosphate-solubilizing bacteria from grapevine rhizospheres for use as biofertilizers. Spanish Journal of Agricultural Research, 14(4), e1106.
Soil science