Skip to main content
SpringerLink
Log in

An improved method for Beauveria bassiana transformation using phosphinothricin acetlytransferase and green fluorescent protein fusion gene as a selectable and visible marker

  • Original Research Paper
  • Published:
Biotechnology Letters Aims and scope Submit manuscript

Abstract

An improved transformation method for the biocontrol agent, Beauveria bassiana, was developed. For convenience of transformation selection and detection, the coding regions of the genes for phosphinothricin acetlytransferase and green fluorescent protein were fused and an expression vector, pBFT, carrying this fusion was constructed. Under optimum conditions, over 60 transformants μg−1 plasmid DNA were obtained. B. bassiana conidia frozen 1 month at −80°C were fully competent for transformation. The method was significantly less laborious and more rapid than current methods for B. bassiana. The bar::egfp provides a selectable and visible marker which may expedite future genetic engineering of this fungus.

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

Similar content being viewed by others

References

  • Cho EM, Boucias D, Keyhani NO (2006a) EST analysis of cDNA libraries from the entomopathogenic fungus Beauveria (Cordyceps) bassiana. II. Fungal cells sporulating on chitin and producing oosporein. Microbiology 152:2855–2864

    Article  PubMed  CAS  Google Scholar 

  • Cho EM, Liu L, Farmerie W et al (2006b) EST analysis of cDNA libraries from the entomopathogenic fungus Beauveria (Cordyceps) bassiana. I. Evidence for stage-specific gene expression in aerial conidia, in vitro blastospores and submerged conidia. Microbiology 152:2843–2854

    Article  PubMed  CAS  Google Scholar 

  • Daboussi MJ, Djeballi A, Gerlinger C et al (1989) Transformation of seven species of filamentous fungi using the nitrate reductase gene of Aspergillus nidulans. Curr Genet 15:453–456

    Article  PubMed  CAS  Google Scholar 

  • Fan Y, Fang W, Guo S et al (2007) Increased insect virulence in Beauveria bassiana strains overexpressing an engineered chitinase. Appl Environ Microbiol 73:295–302

    Article  PubMed  CAS  Google Scholar 

  • Fang W, Zhang Y, Zheng X et al (2004) Agrobacterium tumefaciens-mediated transformation of Beauveria bassiana using an herbicide resistance gene as a selection marker. J Invertebr Pathol 85:18–24

    Article  PubMed  CAS  Google Scholar 

  • Fang W, Leng B, Xiao Y et al (2005) Cloning of Beauveria bassiana chitinase gene Bbchit1 and its application to improve fungal strain virulence. Appl Environ Microbiol 71:363–370

    Article  PubMed  CAS  Google Scholar 

  • Fang W, Pei Y, Bidochka MJ (2006) Transformation of Metarhizium anisopliae mediated by Agrobacterium tumefaciens. Can J Microbiol 52:623–626

    Article  PubMed  CAS  Google Scholar 

  • Jiang Q, Ying SH, Feng MG (2007) Enhanced frequency of Beauveria bassiana blastospore transformation by restriction enzyme-mediated integration and electroporation. J Microbiol Methods 69:512–517

    Article  PubMed  CAS  Google Scholar 

  • Koukaki M, Giannoutsou E, Karagouni A et al (2003) A novel improved method for Aspergillus nidulans transformation. J Microbiol Methods 55:687–695

    Article  PubMed  CAS  Google Scholar 

  • Pfeifer TA, Khachatourians GG (1992) Beauveria bassiana protoplast regeneration and transformation using electroporation. Appl Microbiol Biotechnol 38:376–381

    Article  CAS  Google Scholar 

  • Reader U, Broda P (1985) Rapid preparation of DNA from filamentous fungi. Lett Appl Microbiol 1:17–20

    Article  Google Scholar 

  • Robinson M, Sharon A (1999) Transformation of the bioherbicide Colletotrichum gloeosporioides f. sp. aeschynomene by electroporation of germinated conidia. Curr Genet 36:98–104

    Article  PubMed  CAS  Google Scholar 

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

    Google Scholar 

  • Sánchez O, Aguirre J (1996) Efficient transformation of Aspergillus nidulans by electroporation of germinated conidia. Fungal Genet Newsl 43:48–51

    Google Scholar 

  • Ying SH, Feng MG (2006) Novel blastospore-based transformation system for integration of phosphinothricin resistance and green fluorescence protein genes into Beauveria bassiana. Appl Microbiol Biotechnol 72:206–210

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by the grants from the Ministry of Science and Technology of China (2003CB114203 to Y. Pei) and the Natural Science Foundation of China (30471147 to Y. Zhang).

Author information

Authors and Affiliations

  1. Key Laboratory of Biotechnology and Crop Quality Improvement of Ministry of Agriculture of China, Biotechnology Research Center, Southwest University, Chongqing, 400716, People’s Republic of China

    Kai Jin, Yongjun Zhang, Zhibing Luo, Yuehua Xiao, Yanhua Fan, Di Wu & Yan Pei

Authors
  1. Kai Jin
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yongjun Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Zhibing Luo
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yuehua Xiao
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yanhua Fan
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Di Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Yan Pei
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yan Pei.

Additional information

K. Jin and Y. Zhang contributed equally to this paper.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Jin, K., Zhang, Y., Luo, Z. et al. An improved method for Beauveria bassiana transformation using phosphinothricin acetlytransferase and green fluorescent protein fusion gene as a selectable and visible marker. Biotechnol Lett 30, 1379–1383 (2008). https://doi.org/10.1007/s10529-008-9713-6

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10529-008-9713-6

Keywords

Search

Navigation