Skip to main content

Advertisement

SpringerLink
Log in

Tricalcium phosphate solubilisation by new endophyte Bacillus methylotrophicus CKAM isolated from apple root endosphere and its plant growth-promoting activities

  • Original Paper
  • Published:
Acta Physiologiae Plantarum Aims and scope Submit manuscript

Abstract

Bacillus methylotrophicus CKAM obtained from root endosphere of healthy apple trees was selected on the basis of higher P-solubilisation (687 mg/L), nitrogenase activity (237.6 ηmole C2H4 h−1mg−1 protein), IAA (34 µg/mL), siderophore unit (96.4 %) and antifungal activity against F. oxysporum (88.22 %), Phytophthora sp. (70.00 %), D. necatrix (61.73 %), S. rolfsii (44.54 %) and P. aphanidermatum (62.56 %). We investigated the ability of isolate CKAM to solubilise insoluble P via two possible mechanisms: proton excretion by ammonium assimilation and organic acid production. There were no clear differences in pH and P-solubilisation between glucose–ammonium and glucose–nitrate media. P-solubilisation was significantly promoted with glucose compared with fructose. HPLC study showed that isolate CKAM produced mainly gluconic and oxalic acids with small amounts of 2-ketogluconic, formic acids. During the culture, the pH was reduced with increase in gluconic acid concentration and was inversely correlated with soluble P concentration. Analysis of antifungal compounds involved in their antagonistic activity showed that isolate CKAM produced chitinase, proteases, pectinase and the antibiotic lipopeptides surfactin, fengycin and iturin A. It was notable that isolate CKAM exhibited highest protection against S. rolfsii (58 %) followed by F. oxysporum (54.5 %), D. necatrix (52.7 %), P. aphanidermatum (36.3 %) and Phytophthora sp. (21.8 %) in biocontrol trials using the pathosystem tomato. Remarkable increase was observed in seed germination (27.07 %), shoot length (42.33 %) root length (52.6 %), shoot dry weight (62.01 %) and root dry weight (45.7 %) of tomato under net house condition. Isolate CKAM possessed traits related to plant growth promotion, therefore, could be a potential candidate for the development of biofertiliser or biocontrol agent.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Abdul Babi AA, Anderson JD (1973) Vigour determination in soyabean seed by multiple criteria. Crop Sci 13:630–633

    Article  Google Scholar 

  • Ali B, Sabri AN, Hasnain S (2010) Rhizobacterial potential to alter auxin content and growth of Vigna radiata (L.). World J Microbiol Biotechnol 26:1379–1384

    Article  CAS  Google Scholar 

  • Altschul SF, Thomas LM, Alejandro AS, Jinghui Z (1997) Gapped BLAST and PSIBLAST: a new generation of protein database search programs. Nucl Acids Res 25:3389–3402

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Baker 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 19:451–457

    Article  Google Scholar 

  • Bakthavatchalu S, Shivakumar S, Sullia SB (2012) Identification of multi-trait PGPR isolates and evaluation of their potential as biocontrol agents. Acta Biologica Indica 1:61–67

    Google Scholar 

  • Cazorla FM, Romero D, Garcıa Perez A, Lugtenberg BJJ, de Vicente, Bloemberg G (2007a) Isolation and characterization of antagonistic Bacillus subtilis strains from the avocado rhizoplane displaying biocontrol activity. J Appl Microbiol 103:1950–1959

    Article  CAS  PubMed  Google Scholar 

  • Cazorla FM, Romero D, Garcia AP, Lugtenberg BJJ, Vincente A, Bloemberg G (2007b) Isolation and characterization of antagonistic B. subtilis strains from the avocado rhizoplane displaying biocontrol activity. J Appl Microbiol 103:1950–1959

    Article  CAS  PubMed  Google Scholar 

  • Chatli Anshu S, Beri V, Sidhu BS (2008) Isolation and characterisation of phosphate solubilizing microorganisms from the cold desert habitat of Salix alba Linn. in trans Himalayan region of Himachal Pradesh. Indian J Microbiol 48:267–273

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Chen YP, Rekha PD, Arun AB, Shen FT, Lai WA (2006) Phosphate solubilizing bacteria from subtropical soil and their tri-calcium phosphate solubilizing abilities. Appl Soil Ecol 34:33–41

    Article  Google Scholar 

  • Collins DP, Jacobsen BJ (2003) Optimizing a Bacillus subtilis isolate for biological control of sugar beet cercospora leaf spot. Biol Control 26:153–161

    Article  Google Scholar 

  • Del Campillo SE, Van der Zee S, Torrent J (1999) Modeling long-term phosphorus leaching and changes in phosphorus fertility in excessively fertilized acid sandy soils. Eur J Soil Sci 50:391–399

    Article  Google Scholar 

  • Felsenstein J (1985) Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39:783–791

    Article  Google Scholar 

  • Glick BR (1995) The enhancement of plant growth by free living bacteria. Can J Microbiol 41:109–117

    Article  CAS  Google Scholar 

  • Hardy RWF, Holsten RD, Jacken EK (1968) The acetylene ethylene assay for N2- fixation-laboratory and field evaluation. Plant Physiol 43:118–127

    Google Scholar 

  • Jackson ML (1973) Soil chemical analysis. Prentice Hall, New Delhi, pp 49–95

    Google Scholar 

  • Jeffries S, Gianinazzi S, Perotto S, Turnau K, Barea JM (2003) The contribution of arbuscular mycorhizal fungi in sustainable maintenance of plant health and soil fertility. Biol Fertil Soils 37:1–16

    Google Scholar 

  • Joshi R, McSpadden Gardener B (2006) Identification of genes associated with pathogen inhibition in different strains B. subtilis. Phytopathol 96:145–154

    Article  CAS  Google Scholar 

  • Kumar RS, Ayyadurai N, Pandiaraja P, Reddy AV, Venkateswarlu Y, Sakthivel OPN (2005) Characterization of antifungal metabolite produced by a new strain Pseudomonas aeruginosa PUPa3 that exhibits broad-spectrum antifungal activity and biofertilizing traits. J Appl Microbiol 98:145–154

    Article  CAS  PubMed  Google Scholar 

  • Kumar D, Shivay YS, Dhar S, Kumar C, Prasad R (2013) Rhizospheric flora and the influence of agronomic practices on them—a review. Proc Natl Acad Sci India 83:1–14

    Article  Google Scholar 

  • McSpadden Gardener BB (2004) Ecology of Bacillus and Paenibacillus spp. in agricultural systems. Phytopathol 94:1252–1258

    Article  CAS  Google Scholar 

  • Mehta P, Chauhan A, Mahajan R, Mahajan PK, Shirkot CK (2010) Strain of Bacillus circulans isolated from apple rhizosphere showing plant growth promoting potential. Curr Sci 98:538–542

    CAS  Google Scholar 

  • Mehta P, Walia A, Chauhan A, Shirkot CK (2013) Phosphate solubilization and plant growth promoting potential by stress tolerant Bacillus sp. isolated from rhizosphere of apple orchards in trans Himalayan region of Himachal Pradesh. Ann Appl Biol 163:430–443

    CAS  Google Scholar 

  • Omar SA (1998) The role of rock-phosphate-solubilizing fungi and vesicular-arbuscular-mycorrhiza (VAM) in growth of wheat plants fertilized with rock phosphate. World J Microbiol Biotechnol 14:211–218

    Article  CAS  Google Scholar 

  • Ongena M, Jacques P, Toure Y, Destain J, Jabrane A, Thonart P (2005) Involvement of fengycin-type lipopeptides in the multifaceted biocontrol potential of Bacillus subtilis. Appl Microbiol Biotechnol 69:29–38

    Article  CAS  PubMed  Google Scholar 

  • Park KH, Lee CY, Son HJ (2009) Mechanism of insoluble phosphate solubilization by Pseudomonas fluorescens RAF15 isolated from ginseng rhizosphere and its plant growth promoting activities. Lett Appl Microbiol 49:222–228

    Article  PubMed  Google Scholar 

  • Patel DK, Archana G, Kumar GN (2008) Variation in the nature of organic acid secretion and mineral phosphate solubilization by Citrobacter sp. DHRSS in the presence of different sugars. Curr Microbiol 56:168–174

    Article  CAS  PubMed  Google Scholar 

  • Penrose DM, Glick BR (2003) Methods for isolating and characterizing ACC deaminase-containing plant growth-promoting rhizobacteria. Physiol Plant 118:10–15

    Article  CAS  PubMed  Google Scholar 

  • Powers EM (1995) Efficacy of the Ryu nonstaining KOH technique for rapidly determining gram reactions of foodborne and water-borne bacteria and yeast. Appl Environ Microbiol 61:3758–3765

    Google Scholar 

  • Romero D, Perez-Garcia A, Rivera ME, Cazorla FM, de Vicente A (2004a) Isolation and evaluation of antagonistic bacteria towards the curcurbit powdery mildew fungus Podosphaera fusca. Appl Microbiol Biotechnol 64:263–269

    Article  CAS  PubMed  Google Scholar 

  • Romero D, Perez-Garcia A, Rivera ME, Cazorla FM, de Vicente A (2004b) Isolation and evaluation of antagonistic bacteria towards the curcurbit powdery mildew fungus Podosphaera fusca. Appl Microbiol Biotechnol 64:263–269

    Article  CAS  PubMed  Google Scholar 

  • Romero D, de Vicente A, Rakotoaly RV, Dufour SE, Veening JW, Arrebola E, Cazorla FM, Kuipers OP (2007) The iturin and fengycin families of lipopeptides are key factors in antagonism of Bacillus subtilis towards Podosphaera fusca. Mol Plant Microbe Interact 20:430–440

    Article  CAS  PubMed  Google Scholar 

  • Roos W, Luckner M (1984) Relationships between protonextrusion and fluxes of ammonium ions and organic acid in Penicillium cyclopium. J Gen Microbiol 130:1007–1014

    CAS  Google Scholar 

  • Sasser M, Wichman MD (1991) Identification of microorganisms through use of gas chromatography and high-performance liquid chromatography. In: Balows A, Hausler WJ, Herrman KL, Isenberg HD, Shadomy HJ (eds) Manual of clinical microbiology. American Society for Microbiology, Washington, DC

    Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  • Stein T (2005) Bacillus subtilis antibiotics: structures, syntheses and specific functions. Mol Microbiol 56:845–857

    Article  CAS  PubMed  Google Scholar 

  • Sundara Rao WVB, Sinha MK (1963) Phosphate dissolving microorganisms in the soil and rhizosphere. Indian J Agric Sci 33:272–278

    Google Scholar 

  • Vassilev N, Vassileva M, Nikolaeva I (2006) Simultaneous P-solubilizing and biocontrol activity of microorganisms: potentials and future trends. Appl Microbiol Biotechnol 71:137–144

    Article  CAS  PubMed  Google Scholar 

  • Walia A, Mehta P, Chauhan A, Shirkot CK (2013) Effect of Bacillus subtilis strain CKT1 as inoculum on growth of tomato seedlings under net house conditions. Proc Natl Acad Sci India. doi:10.1007/s40011-013-0189-3

    Google Scholar 

Download references

Acknowledgments

The authors would like to thank ICAR (AINP on Soil Biodiversity & Biofertilizer), New Delhi, India, for providing the financial support.

Author information

Authors and Affiliations

  1. DBT-IOC Centre for Advance Bioenergy Research, Research and Development Centre, Indian Oil Corporation Limited, Sector-13, 121 007, Faridabad, 110067, New Delhi, India

    Preeti Mehta

  2. Forensic Science Laboratory, Sector-14, Rohini, 110085, India

    Abhishek Walia

  3. Analytical Department, Jubilant Chemsys Limited, B-34 Sector-58, Noida, 20130, India

    Nitin Kakkar

  4. Department of Basic Sciences, Dr. Y. S. Parmar University of Horticulture and Forestry, Nauni, Solan, 173230, Himachal Pradesh, India

    C. K. Shirkot

Authors
  1. Preeti Mehta
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Abhishek Walia
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Nitin Kakkar
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. C. K. Shirkot
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Preeti Mehta.

Additional information

Communicated by A. Krolicka.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Mehta, P., Walia, A., Kakkar, N. et al. Tricalcium phosphate solubilisation by new endophyte Bacillus methylotrophicus CKAM isolated from apple root endosphere and its plant growth-promoting activities. Acta Physiol Plant 36, 2033–2045 (2014). https://doi.org/10.1007/s11738-014-1581-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11738-014-1581-1

Keywords

Search

Navigation