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Only matings between individuals of different mating type are allowed.
The Leupold strain 968 (h mating type) corresponds to CBS1042, whereas Leupold strains 972 (h -S mating type) and 975 (h +N mating type) are spontaneously occurring heterothallic derivatives of CBS1042 (Munz et al. 1989).
The majority of natural isolates are haploid, but they possess one of two mating type alleles, α or a, and in the laboratory, strains of opposite mating type are capable of sexual reproduction [11].
Further studies with genotyping by pulsed field gel electrophoresis and mating type are needed.
In a classical breeding strategy, haploid strains of opposite mating type are crossed to produce new diploid progeny.
Thus, simple, rapid, and efficient methods for bulk isolation of pure ascospores sorted by mating type are needed.
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Only one stable haploid derivative (4124-S60) was isolated, while 14 other segregants with a stable mating type were aneuploid.
Finally the engineered haploids with opposite mating type were mated back to diploid strains, obtaining a heterozygous deletion strain (CAR1/car1) and a homozygous defect strain (car1/car1).
A significant bias towards the self-fertile mating type was observed in all single ascospore masses taken from sexual structures produced in haploid-selfed cultures.
Strains of opposite mating type were mixed in equal amount for the heterothallic a-α mating assay, otherwise, cell suspensions of single mating type strain were directly conducted for the same-sex mating assay.
Specifically, 10 isolates with identical genotypes and the MATa mating type were isolated from the Tu422 site, which was associated with a mopane tree (Table S1).
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