Role of bacterial virulence proteins in Agrobacterium-mediated transformation of Aspergillus awamori.
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.
Michielse
C B
CB
Institute of Biology, Clusius Laboratory, Leiden University, Wassenaarseweg 64, 2333 AL, Leiden, The Netherlands. michielse@rulbim.leidenuniv.nl
Ram
A F J
AF
Hooykaas
P J J
PJ
Hondel
C A M J J van den
CA
D016960:Agrobacterium tumefaciens / Q000235:genetics* / Q000502:physiology*; D001230:Aspergillus / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics / Q000502:physiology; D003227:Conjugation, Genetic; D004269:DNA, Bacterial / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics / Q000502:physiology; D017353:Gene Deletion; D005798:Genes, Bacterial; D007473:Ion Channels / Q000235:genetics / Q000502:physiology; D016254:Mutagenesis, Insertional; D036741:Plant Tumor-Inducing Plasmids; D013696:Temperature; D013997:Time Factors; D014170:Transformation, Genetic; D037521:Virulence Factors / Q000235:genetics / Q000502:physiology
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D007473:Ion Channels; C036483:T-DNA; D037521:Virulence Factors; C051583:virE2 protein, Agrobacterium