Agrobacterium tumefaciens-mediated genetic transformation of the entomopathogenic fungus Beauveria bassiana

Journal of Microbiological Methods
Maria Cecília dos ReisM C Furlaneto

Abstract

Agrobacterium tumefaciens-mediated transformation (agro-transformation) was successfully applied to the entomogenous fungus Beauveria bassiana. Conidia of B. bassiana were transformed to hygromycin B resistance using the hph gene of Escherichia coli as the selective trait, under the control of a heterologous fungal promoter and the Aspergillus nidulans trpC terminator. The efficiency of transformation was up to 28 and 96 transformants per 10(4) and 10(5) target conidia, respectively, using three distinct vectors. High mitotic stability of the transformants (80-100%) was demonstrated after five successive transfers on a nonselective medium. Abortive transformants were observed for all the hph(r) vectors used. Putative transformants were analysed for the presence of the hph gene by PCR and Southern analysis. The latter analysis revealed the integration of two or more copies of the hph gene in the genome. The agro-transformation method was found to be effective for the isolation of B. bassiana hygromycin resistant transformants and may represent a useful tool for insertional mutagenesis studies in this fungus.

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Citations

Jul 19, 2012·World Journal of Microbiology & Biotechnology·Liang ShiMingwen Zhao
Jun 7, 2005·Journal of Microbiological Methods·Aída V Rodríguez-TovarBeatriz Xoconostle-Cázares
May 13, 2005·Current Genetics·Caroline B MichielseArthur F J Ram
Dec 10, 2009·Journal of Microbiological Methods·J G FigueiredoC Glienke
Apr 25, 2008·Bioresource Technology·Jie WangJunfang Lin
May 19, 2006·Letters in Applied Microbiology·I G P LimaM C Furlaneto
Nov 18, 2005·Trends in Genetics : TIG·Benoît LacroixVitaly Citovsky
Aug 19, 2005·Journal of Microbiological Methods·Richard J WeldHayley J Ridgway
Aug 16, 2015·Fungal Genetics and Biology : FG & B·Changwen ShaoZhongkang Wang
Aug 13, 2013·Fungal Biology·Maria Beatriz Calderan RodriguesAline Aparecida Pizzirani-Kleiner
Apr 14, 2009·Methods in Enzymology·Russell J Cox, Thomas J Simpson
Feb 10, 2006·Cell Research·Richard J WeldHayley J Ridgway
May 15, 2007·Canadian Journal of Microbiology·Marcia M MataMaria Helena P Fungaro
Sep 8, 2017·Scientific Reports·Pamil TayalAtul Kumar Johri
Feb 26, 2019·3 Biotech·Kawkab KanjoGurvinder Kaur Saini
Nov 18, 2018·Molecular Genetics and Genomics : MGG·Inoka K HettiarachchigeGerman C Spangenberg

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