Studying the mating system in Botrytis cinerea

iv Introduction 1 Sex in fungi 1 Mating systems and MAT loci in ascomycetous fungi 2 Botrytis cinerea Pers. Fr.(teleomorph: Botryotinia fuckelinia [de Bary] Whetz.) 3 Sexual fruiting bodies: "Apothecia" 3 Aberrant mating behavior of Botrytis cinerea 4 Materials and Methods 5 Botrytis cinerea growth conditions for DNA isolation 5 Botrytis cinerea growth conditions for mating experiments 5 Sexual crosses 6 MAT loci amplification 6 Checking MAT idiomorphs using PCR 7 Target mutagenesis of genes in the MAT loci of B. cinerea 7 Designing primers for gene replacement 7 Amplification of constructs 8 Protoplast formation 9 Transformation 10 Screening for targeted mutagenesis. 10 Results 11 Sequence analysis of the MAT locus in B. cinerea strains 11 Mating behavior and MAT locus analysis of different B. cinerea strains. 12 Cross of dual mater strain RS26 with MAT 1-1 reference strain 12 Cross of dual mater strain RS26 with MAT 1-2 reference strain 12 Cross of MAT1-1 reference SAS 56 with MAT1-2 reference SAS 405 13 Self-fer le strain Bc16 and its progeny 15 Targeted mutagenesis of genes in the MAT loci of B. cinerea strains 15 Discussion 17 Mating behavior and MAT loci analysis of different B. cinerea strains 17 Sequence analysis of the MAT loci of B. cinerea strains. 18 Targeted mutagenesis of genes in the MAT locus 19