title: Identification of pathogenesis-associated genes by T-DNA-mediated insertional mutagenesis in Botrytis cinerea: a type 2A phosphoprotein phosphatase and an SPT3 transcription factor have significant impact on virulence.
abstract: Agrobacterium tumefaciens-mediated transformation (ATMT) was used to generate an insertional mutant library of the gray mold fungus Botrytis cinerea. From a total of 2,367 transformants, 68 mutants showing significant reduction in virulence on tomato and bean plants were analyzed in detail. As reported for other fungal ATMT libraries, integrations were mostly single copy, occurred preferentially in noncoding (regulatory) regions, and were frequently accompanied by small deletions of the target sequences and loss of parts of the border sequence. Two T-DNA integration events that were found to be linked to virulence were characterized in more detail: a catalytic subunit of a PP2A serine/threonine protein phosphatase (BcPP2Ac) and the SPT3 subunit of a Spt-Ada-Gcn5-acetyltransferase (SAGA-like) transcriptional regulator complex. Gene replacement and silencing approaches revealed that both Bcpp2Ac and SPT3 are crucial for virulence, growth, and differentiation as well as for resistance to H(2)O(2) in B. cinerea.
authors: Institut für Biologie und Biotechnologie der Pflanzen, Westf. Wilhelms-Universität, Hindenburgplatz 55, D-48143 Münster, Germany.
authors: S
authors: S
authors: Giesbert
authors: J
authors: J
authors: Schumacher
authors: V
authors: V
authors: Kupas
authors: J
authors: J
authors: Espino
authors: N
authors: N
authors: Segmüller
authors: I
authors: I
authors: Haeuser-Hahn
authors: P H
authors: PH
authors: Schreier
authors: P
authors: P
authors: Tudzynski
mesh_terms: D020171:Botrytis / Q000235:genetics* / Q000378:metabolism* / Q000472:pathogenicity; D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism*; D015966:Gene Expression Regulation, Fungal / Q000502:physiology; D015723:Gene Library; D020868:Gene Silencing; D016254:Mutagenesis, Insertional; D027805:Phaseolus / Q000382:microbiology; D010749:Phosphoprotein Phosphatases / Q000235:genetics / Q000378:metabolism*; D010935:Plant Diseases / Q000382:microbiology; D060888:Real-Time Polymerase Chain Reaction; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014774:Virulence
chemical_list: D004269:DNA, Bacterial; C036483:T-DNA; D014157:Transcription Factors; D010749:Phosphoprotein Phosphatases