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.

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SUMMARYGene flow from herbicide-resistant rice (Oryza sativaL.) cultivars can affect the biodiversity ofOryzaspp. and can result in the lack of opportunity to control weedy rice through selective herbicides. The aim of the present study was to quantify the outcrossing rate from the herbicide-resistant red rice and rice cultivars carrying three different ALS (acetolactate synthase) alleles using a single plant pollen donor approach. A field experiment was performed using the encircle population combination technique. The main plots comprised the pollen-receptor IRGA 417 cultivar or a susceptible biotype of weedy rice, and the subplots comprised the pollen-donor inbred cultivars IRGA 422 CL and PUITÁ INTA CL, the hybrid SATOR CL or a resistant biotype of weedy rice. Among the pollen-donors, the outcrossing rate for pollen receptor susceptible weedy rice and the IRGA 417 cultivar was 0.0344% and 0.0142%, respectively. Rice cultivars carrying theALSgene mutations Ala122Thr, Ser653Asn and Gly654Asn showed a similar outcrossing rate of 0.0243%. The outcrossing rate decreased over a distance of up to 3.5 m from the pollen-donor and was not affected by the wind cardinal direction. The risk of gene flow of herbicide resistance from rice to weedy rice should be reduced through the development of new strategies to contain and mitigate gene flow and of the elimination of weedy rice escapees.

RESUMO Diversos biótipos de sagitária (Sagittaria montevidensis Cham. & Schlecht) foram constatados como resistentes a herbicidas inibidores da enzima ALS no Brasil e em outros locais do mundo. Os objetivos deste trabalho foram

RESUMO Diversos biótipos de sagitária (Sagittaria montevidensis Cham. & Schlecht) foram constatados como resistentes a herbicidas inibidores da enzima ALS no Brasil e em outros locais do mundo. Os objetivos deste trabalho foram