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Role of bacterial virulence proteins in Agrobacterium-mediated transformation of Aspergillus awamori

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Author: Michielse, C.B. · Ram, A.F.J. · Hooykaas, P.J.J. · Hondel, C.A.M.J.J. van den
Type:article
Date:2004
Institution: TNO Voeding
Source:Fungal Genetics and Biology, 5, 41, 571-578
Identifier: 237732
doi: doi:10.1016/j.fgb.2004.01.004
Keywords: Biology · Biotechnology · bacterial protein · protein VirA · protein virC · protein virD · protein virG · regulator protein · unclassified drug · article · Aspergillus awamori · bacterial virulence · biotransformation · controlled study · cultivar · DNA strand · DNA transfer · host pathogen interaction · nonhuman · plant · priority journal · protein targeting · reaction optimization · reduction · reproducibility · Rhizobium · system analysis · Aspergillus · Bacterial Proteins · Conjugation, Genetic · DNA, Bacterial · DNA-Binding Proteins · Gene Deletion · Genes, Bacterial · Ion Channels · Mutagenesis, Insertional · Plant Tumor-Inducing Plasmids · Rhizobium radiobacter · Temperature · Time Factors · Transformation, Genetic · Virulence Factors · Agrobacterium · Agrobacterium tumefaciens · Aspergillus · Aspergillus awamori · Bacteria (microorganisms) · Viruses (infective agents)

Abstract

The Agrobacterium-mediated transformation of Aspergillus awamori was optimized using defined co-cultivation conditions, which resulted in a reproducible and efficient transformation system. Optimal co-cultivation conditions were used to study the role of Agrobacterium tumefaciens virulence proteins in T-DNA transfer. This study revealed that inactivation of either of the regulatory proteins (VirA, VirG), any of the transport pore proteins (VirB), proteins involved in generation of the T-strand (VirD, VirC) or T-strand protection and targeting (VirE2) abolishes or severely reduces the formation of transformants. The results indicate that the Agrobacterium-mediated transformation of A. awamori requires an intact T-DNA machinery for efficient transformation; however, the plant host range factors, like VirE3, VirH, and VirF, are not important. Index descriptors: Agrobacterium tumefaciens; T-DNA; Virulence proteins; DNA-transformation; Aspergillus awamori; Filamentous fungi © 2004 Elsevier Inc. All rights reserved.