HIGH-EFFICIENCY TRANSFORMATION OF THE PLANT PATHOGENIC FUNGUS MARSSONINA BRUNNEA

C. Jiang, X. Cheng, Q. Cheng, M. Huang, L. Xu

Abstract


Marssonina brunnea is the causal agent of Marssonina leaf spot of poplar. We successfully transformed M. brunnea using Agrobacterium tumefaciens. A highly-efficient promoter was a key factor in the construction of this transformation system construction. The beta -tubulin gene is stable and highly expressed at almost all stages of the life history of M. brunnea. A β-tubulin gene promoter was obtained based on a genomic sequence using an adapter ligation-mediated PCR. This was then inserted into the pMDC83 plasmid vector for high expression of the inserted gene. A genomic DNA PCR assay and fluorescence observation of transformants authenticated successful transformation and fluorescence intensity indicated the efficacy of the cloned promoter. All M. brunnea transformants were mitotically stable. This study provides essential information for the study of the functions of virulence genes and their roles in pathogen-host interactions.

Keywords


Agrobacterium tumefaciens-mediated transformation; full length cDNA cloning; green fluorescent protein expression; promoter cloning

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DOI: http://dx.doi.org/10.4454/JPP.V96I3.005

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EDIZIONI ETS, Pisa, Italy