Skip to main content
SpringerLink
Log in

Isolation of Oxalic acid tolerating fungi and decipherization of its potential to control Sclerotinia sclerotiorum through oxalate oxidase like protein

  • Original Paper
  • Published:
World Journal of Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

Oxalic acid plays major role in the pathogenesis by Sclerotinia sclerotiorum; it lowers the pH of nearby environment and creates the favorable condition for the infection. In this study we examined the degradation of oxalic acid through oxalate oxidase and biocontrol of Sclerotinia sclerotiorum. A survey was conducted to collect the rhizospheric soil samples from Indo-Gangetic Plains of India to isolate the efficient fungal strains able to tolerate oxalic acid. A total of 120 fungal strains were isolated from root adhering soils of different vegetable crops. Out of 120 strains a total of 80 isolates were able to grow at 10 mM of oxalic acid whereas only 15 isolates were grow at 50 mM of oxalic acid concentration. Then we examined the antagonistic activity of the 15 isolates against Sclerotinia sclerotiorum. These strains potentially inhibit the growth of the test pathogen. A total of three potential strains and two standard cultures of fungi were tested for the oxalate oxidase activity. Strains S7 showed the maximum degradation of oxalic acid (23 %) after 60 min of incubation with fungal extract having oxalate oxidase activity. Microscopic observation and ITS (internally transcribed spacers) sequencing categorized the potential fungal strains into the Aspergillus, Fusarium and Trichoderma. Trichoderma sp. are well studied biocontrol agent and interestingly we also found the oxalate oxidase type activity in these strains which further strengthens the potentiality of these biocontrol agents.

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

  • Aguiar A, Brazil de Souza-Cruz P, Ferraz A (2006) Oxalic acid, Fe3+ reduction activity and oxidative enzymes detected in culture extracts recovered from Pinus taeda wood chips biotreated by Ceriporiopsis subvermispora. Enzyme Microb Technol 38:873–878

    Article  CAS  Google Scholar 

  • Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W, Lipman OJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402

    Article  CAS  Google Scholar 

  • Bateman DF, Beer SV (1965) Simultaneous production and synergistic action of oxalic acid and polygalacturonase during pathogenesis by Sclerotiorum rolfsii. Phytopathology 55:204–211

    CAS  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  Google Scholar 

  • Burke JM, Rieseberg LH (2003) Fitness effects of transgenic disease resistance in sunflowers. Science 300:1250

    Article  CAS  Google Scholar 

  • Cotton P, Kasza Z, Bruel C, Rascle C, Fevre M (2003) Ambient pH controls the expression of endopolygalacturonase genes in the necrotrophic fungus Sclerotinia sclerotiorum. FEMS Microbiol Lett 227:163–169

    Article  CAS  Google Scholar 

  • Donaldson PA, Andersonb T, Lanec BG, Davidsona AL, Simmondsa DH (2001) Soybean plants expressing an active oligomeric oxalate oxidase from the wheat gf-2.8 (germin) gene are resistant to the oxalate-secreting pathogen Sclerotinia sclerotiorum. Physiol Mol Plant Pathol 59:297–307

    Article  CAS  Google Scholar 

  • Dumas B, Freyssinet C, Pallett KE (1995) Tissue-Specific Expression of Cermin-Like Oxalate Oxidase during Development and Fungal lnfection of Barley Seedlings. Plant Physiol 107:1091–1096

    CAS  Google Scholar 

  • Escutia MR, Bowater L, Edwards A, Bottrill AR, Burrell MR, Polanco R, Vicuna R, Bornemann S (2005) Cloning and Sequencing of Two Ceriporiopsis subvermispora Bicupin Oxalate Oxidase Allelic Isoforms: implications for the Reaction Specificity of Oxalate Oxidases and Decarboxylases. Appl Environ Microbiol 71:3608–3616

    Article  CAS  Google Scholar 

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

    Article  Google Scholar 

  • Franceschi VR, Nakata PA (2005) Calcium oxalate in plants: formation and function. Ann Rev Plant Biol 56:41–71

    Article  CAS  Google Scholar 

  • Godoy G, Steadman JR, Dickman MB, Dam R (1990) Use of mutants to demonstrate the role of oxalic acid in pathogenicity of Sclerotinia sclerotiorum on Phaseolus vulgaris. Physiol Mol Plant Pathol 37:179–191

    Article  CAS  Google Scholar 

  • Graz M, Wilkołazka AJ, Pawlega PB (2009) Abortiporus biennis tolerance to insoluble metal oxides: oxalate secretion, oxalate oxidase activity, and mycelia morphology. Biometals 22:401–410

    Article  CAS  Google Scholar 

  • Grąz M, Jarosz-Wilkołazka A, Pawlikowska-Pawlęga B (2009) Abortiporus biennis tolerance to insoluble metal oxides: oxalate secretion, oxalate oxidase activity, and mycelia morphology. Biometals 22:401–410

    Article  Google Scholar 

  • Hamel R, Levasseur R, Appanna VD (1999) Oxalic acid production and aluminum tolerance in Pseudomonas fluorescens. J Inorg Biochem 76:99–104

    Article  CAS  Google Scholar 

  • Jarosz-Wilkołazka A, Graz M (2006) Organic acids production by white rot Basidiomycetes in the presence of metallic oxides. Can J Microbiol 52:779–785

    Article  Google Scholar 

  • Kesarwani M, Azam M, Natarajan K, Mehta A, Datta A (2000) Oxalate decarboxylase from Collybia velutipes. Molecular cloning and its overexpression to confer resistance to fungal infection in transgenic tobacco and tomato. J Biol Chem 275:7230–7238

    Article  CAS  Google Scholar 

  • Libert B, Franceschi VR (1987) Oxalate in crop plants. J Agric Food Chem 35:926–938

    Article  CAS  Google Scholar 

  • Liu E–E, Luo W, Zhou H, Pen X–X (2009) Determination of oxalate in plant tissues with oxalate oxidase prepared from wheat. Biol Plant 53:129–132

    Article  CAS  Google Scholar 

  • Marciano P, di Lenna P, Margo P (1983) Oxalic acid, cell-wall degrading enzymes and pH in pathogenesis and their significance in the virulence of two Sclerotinia sclerotiorum isolates on sunflower. Physiol Plant Pathol 22:339–345

    CAS  Google Scholar 

  • Maxwell DP, Bateman DF (1968) Oxalic acid biosynthesis by Sclerotium rolfsii. Phytopathology 58:1635–1642

    CAS  Google Scholar 

  • Nakata PA, He C (2010) Oxalic acid biosynthesis is encoded by an operon in Burkholderia glumae. FEMS Microbiol Lett 304:177–182

    Article  CAS  Google Scholar 

  • Punja ZK, Jenkins SF (1984) Light and scanning electron microscopic observations of calcium oxalate crystals produced during growth of Sclerotium rolfsii in culture and infected tissue. Can J Bot 62:2028–2032

    Article  CAS  Google Scholar 

  • Reinhardt LA, Svedruzic D, Chang CH, Cleland WW, Richards NGJ (2003) Heavy atom isotope effects on the reaction catalyzed by the oxalate decarboxylase from Bacillus subtilis. J Am Chem Soc 125:1244–1252

    Article  CAS  Google Scholar 

  • Ren L, Li G, Han YC, Jiang DH, Huang H-C (2007) Degradation of oxalic acid by Coniothyrium minitans and its effects on production and activity of β-1,3-glucanase of this mycoparasite. Biol Control 43:1–11

    Article  CAS  Google Scholar 

  • Ren L, Li G, Jiang D (2009) Characterization of some culture factors affecting oxalate degradation by the mycoparasite Coniothyrium minitans. J Appl Microbiol 108:173–180

    Article  Google Scholar 

  • Requena L, Bornemann S (1999) Barley (Hordeum vulgare) oxalate oxidase is a manganese-containing enzyme. Biochem J 343:185–190

    Article  CAS  Google Scholar 

  • Rollins JA, Dickman MB (2001) pH signaling in Sclerotinia sclerotiorum: Identification of a pacC/RIM1 homolog. Appl Environ Microbiol 67:75–81

    Article  CAS  Google Scholar 

  • Saitou N, Nei M (1987) The neighbor-joining method: A new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425

    CAS  Google Scholar 

  • Schmid J, Muller-Hagen D, Bekel T, Funk L, Stahl U, Sieber V, Meyer V (2010) Transcriptome sequencing and comparative transcriptome analysis of the scleroglucan producer Sclerotium rolfsii. BMC Genomics 11:329

    Article  Google Scholar 

  • Shannon CE, Weaver W (1949) The mathematical theory of communication. University of Illinois Press, Urbana

    Google Scholar 

  • Singh UP, Sarma BK, Singh DP (2003) Effect of plant growth promoting rhizobacteria and culture filtrate of Sclerotium rolfsii on phenolic and salicylic acid contents in chickpea (Cicer arietinum). Curr Microbiol 46:131–140

    Article  CAS  Google Scholar 

  • Svedruzic D, Jonsson S, Toyota CG, Reinhardt LA, Ricagno S, Lindqvist Y, Richards NGJ (2005) The enzymes of oxalate metabolism: unexpected structures and mechanism. Arch Biochem Biophys 433:176–192

    Article  CAS  Google Scholar 

  • Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol Biol Evolution 24:1596–1599

    Article  CAS  Google Scholar 

  • Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving sensitivity of progressive multiple sequence alignments through sequence weighing, position specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–7680

    Article  CAS  Google Scholar 

  • Watanabe T, Hattori T, Tengku S, Shimada M (2005) Purification and characterization of NAD-dependent formate dehydrogenase from the white-rot fungus Ceriporiopsis subvermispora and a possible role of the enzyme in oxalate metabolism. Enzyme Microb Technol 37:68–75

    Article  CAS  Google Scholar 

  • White TJ, Bruns T, Lee S, Taylor JW (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds) PCR protocols: a guide to methods and applications. Academic Press Inc., New York, pp 315–322

    Google Scholar 

  • Zhou T, Boland GJ (1999) Mycelial growth and production of oxalic acid by virulent and hypovirulent isolates of Sclerotinia sclerotiorum. Can J Plant Pathol 21:93–99

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by Indian Council of Agricultural Research, India, Network Project on “Application of Microorganisms in Agriculture and Allied Sectors”.

Author information

Authors and Affiliations

  1. National Bureau of Agriculturally Important Microorganisms, Mau Nath Bhanjan, Uttar Pradesh, 275101, India

    Shivani Yadav, Alok K. Srivastava, Dhanajay P. Singh & Dilip K. Arora

Authors
  1. Shivani Yadav
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Alok K. Srivastava
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Dhanajay P. Singh
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Dilip K. Arora
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Shivani Yadav.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Yadav, S., Srivastava, A.K., Singh, D.P. et al. Isolation of Oxalic acid tolerating fungi and decipherization of its potential to control Sclerotinia sclerotiorum through oxalate oxidase like protein. World J Microbiol Biotechnol 28, 3197–3206 (2012). https://doi.org/10.1007/s11274-012-1130-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11274-012-1130-2

Keywords

Search

Navigation