Skip to main content
Log in

Development of Strain-specific Primers for a Strain of Gliocladium catenulatum Used in Biological Control

  • Published:
European Journal of Plant Pathology Aims and scope Submit manuscript

Abstract

The randomly amplified polymorphic DNA (RAPD) technique was used to develop strain-specific primers for Gliocladium catenulatum strain J1446, which is promising in biological control. One of the primer pairs developed proved to be strain-specific; strain J1446 was differentiated from 16 G. catenulatum strains and six other strains of two Gliocladium species, as well as from Trichoderma virens, and isolates of Nectria spp. and Fusarium spp. Specific primers were also tested with DNA isolated from cucumber leaves, treated or untreated with a solution made from Gliocladium powder. The expected amplification product was produced only from treated leaves. DNA isolated from Gliocladium-treated potato tubers and fungi grown in peat was also used in amplification reactions. Strain-specific primers detected strain J1446 when the amount of DNA was 5 pg or more. Some variation between the Gliocladium strains was found by the random amplified microsatellites method (RAMS) and the universally primed polymerase chain reaction method (UP-PCR), but no clear fragments specific to strain J1446 were produced. Cross-blot hybridisation of UP-PCR products differentiated strain J1446 from T. virens, but not from the Gliocladium isolates. The 28S rDNA sequences and β-tubulin sequences were identical or very similar in all Gliocladium strains. Thus, it is possible that the Gliocladium strains of the present study are conspecific, which means that a revision in the taxonomy of Gliocladium species may be necessary.

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

Access this article

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

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Bulat SA, Lübeck M, Alekhina IA, Jensen DF and Lubeck PS (1998) UP-PCR analysis and ITS1 ribotyping of Trichoderma and Gliocladium fungi. Mycol Res 102: 933–943

    Google Scholar 

  • Burt A, Carter DA, White TJ and Taylor JW (1994) DNA sequencing with arbitrary primer pairs. Mol Ecology 3: 523–525

    Google Scholar 

  • Doohan FM, Parry DW, Jenkinson P and Nicholson P (1998) The use of species-specific PCR-based assays to analyse Fusarium ear blight of wheat. Plant Pathol 47: 197–205

    Google Scholar 

  • Hantula J, Dusabenyagasani M and Hamelin RC (1996) Random amplified microsatellites (RAMS)-a novel method for characterizing genetic variation within fungi. Eur J For Pathol 26: 159–166

    Google Scholar 

  • Knudsen IMB, Hockenhull J, Jensen DF, Gerhardson B, Hökeberg M, Tahvonen R, Teperi E, Sundheim L and Henriksen B (1997) Selection of biological control agents for controlling soil and seed-borne diseases in the field. Eur J Plant Pathol 103: 775–784

    Google Scholar 

  • Lübeck M (1997) Marker genes and PCR based approaches as tools for monitoring fungi in soil with emphasis on UP-PCR and fungi from the genera Trichoderma and Gliocladium. Ph.D. 198 Thesis, The Royal Veterinary and Agricultural University, Copenhagen, Denmark

    Google Scholar 

  • O'Donnell K and Cigelnik E (1997) Two divergenic intragenomic rDNA ITS2 types within a monophyletic lineage of the fungus Fusarium are nonorthologous. Mol Phylogen Evol 7: 103–116

    Google Scholar 

  • Paavanen-Huhtala S, Hyvönen J, Bulat SA and Yli-Mattila T (1999) RAPD-PCR, isozyme, rDNA RFLP and rDNA sequence analyses in identification of Finnish Fusarium oxysporum isolates. Mycol Res 103: 625–634

    Google Scholar 

  • Papavizas GC (1985) Trichoderma and Gliocladium: biology, ecology and potential for biocontrol. Ann Rev Phytopathology 23: 23–54

    Google Scholar 

  • Paran I and Michelmore RW (1993) Development of reliable PCR-based markers linked to downy mildew resistance genes in lettuce. Theor Appl Genet 85: 985–993

    Google Scholar 

  • Pearson WR and Lipman DJ (1988) Improved tools for biological sequence comparison. Proc Natl Acad Sci USA 85: 2444–2448

    Google Scholar 

  • Rehner SA and Samuels GJ (1994) Taxonomy and phylogeny of Gliocladium analysed from nuclear large subunit ribosomal DNA sequences. Mycol Res 94: 625–634

    Google Scholar 

  • Sambrook J, Fritsch EF and Maniatis T (1989) Molecular Cloning: A Laboratory Manual. 2nd edn. Cold Spring Harbor Laboratory Press, New York

    Google Scholar 

  • Schilling AG, Möller EM and Geiger HH (1996) Polymerase chain reaction-based assays for species-specific detection of Fusarium culmorum, F. graminearum, and F. avenaceum. Phytopathology 86: 515–522

    Google Scholar 

  • Schroers H-J, Samuels GJ, Seifert KA and Gams W (1999) Classification of the mycoparasite Gliocladium roseum in Clonostachys as C. rosea, its relationship to Bionectria ochroleuca, and notes on other Gliocladium-like fungi. Mycol 91: 365–385

    Google Scholar 

  • Teperi E, Keskinen M, Ketoja E and Tahvonen R (1998) Screening for fungal antagonists of seed-borne Fusarium culmorum on wheat using in vivo tests. Eur J Plant Pathol 104: 243–251

    Google Scholar 

  • Thompson JD, Higgins DG and Gibson TJ (1994) Clustal W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22: 4673–4680

    Google Scholar 

  • Ward E and Adams MJ (1998) Analysis of ribosomal DNA sequences of Polymyxa species and related fungi and the development of genus-and species-specific PCR primers. Mycol Res 102: 965–974

    Google Scholar 

  • White TJ, Bruns T, Lee S and Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ and White TJ (eds) PCR Protocols: A guide to Methods and Applications. Academic Press, New York

    Google Scholar 

  • Yli-Mattila T, Paavanen S, Hannukkala A, Parikka P, Tahvonen R and Karjalainen R (1996) Isozyme and RAPD-PCR analyses of Fusarium avenaceum strains from Finland. Plant Pathol 45: 126–134

    Google Scholar 

  • Yli-Mattila T, Mironenko NV, Alekhina I, Hannukkala A, Hyvönen J and BulatSA(1998) Identification of Fusarium avenaceum isolates by RAPD-PCR and UP-PCR. In: Manceau C and Spak J (eds) Advances in the Detection of Plant Pathogens by Polymerase Chain Reaction, (pp 114–123) Proceedings of COST 823 Workshop Ceske Budejovice, 20–21 June, 1996

  • Yli-Mattila T, Paavanen-Huhtala S, Avikainen H and Teperi E (1997) Molecular identification of Gliocladium and Nectria isolates in Finland. Cer Res Comm 25: 293–297

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Paavanen-Huhtala, S., Avikainen, H. & Yli-Mattila, T. Development of Strain-specific Primers for a Strain of Gliocladium catenulatum Used in Biological Control. European Journal of Plant Pathology 106, 187–198 (2000). https://doi.org/10.1023/A:1008734519852

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1008734519852

Navigation