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

Autor: S. Giesbert, J. Schumacher, V. Kupas, J. Espino, N. Segmüller, I. Haeuser-Hahn, P. H. Schreier, P. Tudzynski
Jazyk: angličtina
Rok vydání: 2012
Předmět:
Zdroj: Molecular Plant-Microbe Interactions, Vol 25, Iss 4, Pp 481-495 (2012)
Druh dokumentu: article
ISSN: 1943-7706
0894-0282
DOI: 10.1094/MPMI-07-11-0199
Popis: 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 H2O2 in B. cinerea.
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