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Identification of three mutant loci conferring carboxin-resistance and development of a novel transformation system in Aspergillus oryzae

Identification of three mutant loci conferring carboxin-resistance and development of a novel transformation system in Aspergillus oryzae,10.1016/j.fg

Identification of three mutant loci conferring carboxin-resistance and development of a novel transformation system in Aspergillus oryzae   (Citations: 10)
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Mutants exhibiting resistance to the fungicide, carboxin, were isolated from Aspergillus oryzae, and the mutations in the three gene loci, which encode succinate dehydrogenase (SDH) B, C, and D subunits, were identified to be independently responsible for the resistance. A structural model of the SDH revealed the different mechanisms that confer carboxin-resistance in different mutations. The mutant AosdhB gene (AosdhBcxr) was further examined for possible use as a transformant selection marker. After transformation with AosdhBcxr, carboxin-resistant colonies appeared within 4 days of culture, and all of the examined colonies carried the transgene. Insertion analyses revealed that the AosdhBcxr gene was integrated into AosdhB locus via homologous recombination at high efficiency. Furthermore, AosdhBcxr functioned as a successful selection marker in a transformation experiment in Aspergillus parasiticus, suggesting that this transformation system can be used for Aspergillus species.
Journal: Fungal Genetics and Biology - FUNGAL GENET BIOL , vol. 46, no. 1, pp. 67-76, 2009
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    • ...Other mutations leading to an amino acid substitution in the SdhB, SdhC and SdnD were responsible for laboratory resistance to boscalid analogue-fungicides in some bacteria and basidiomycete and ascomycete fungi (Broomfield and Hargreaves 1992; Ito et al. 2004; Keon et al. 1991; Li et al. 2006; Matsson and Hederstedt 2001; Matsson et al. 1998; Shima et al. 2008; Skinner et al. 1998)...
    • ...The sequences obtained contained the flanking regions from the second (S2) to third cysteine-rich (S3) center of the subunit (Fig. 1). The substitution of a proline residue in the S2 center (Stammler et al. 2008) and a histidine residue in the S3 center (Avenot et al. 2008; Broomfield and Hargreaves 1992; Keon et al. 1991; Matsson and Hederstedt 2001; Shima et al. 2008; Skinner et al. 1998) have been reported to be responsible for reduced ...
    • ...Mutants derived from UV mutagenesis in Aspergillus oryzae also had several levels of sensitivity to carboxin (Shima et al. 2008)...

    Takuya Miyamotoet al. Occurrence of boscalid resistance in cucumber powdery mildew in Japan ...

    • ...solution (Sanderson and Srb 1965), 5 g sodium acetate, 0.01 M citric acid, adjusted at pH 6.0 with KOH]...

    Rita M. De Miccolis Angeliniet al. Selection, characterization and genetic analysis of laboratory mutants...

    • ...In fact, a mutated sdhB gene encoding the Ip subunit has been employed as a selective marker in Pleurotus ostreatus (Honda et al. 2000; Irie et al. 1998), Lentinula edodes (Irie et al. 2003), and Aspergillus oryzae (Shima et al. 2008); however, there has been no report of a transformation system using a homologous gene, a mutated sdhB gene, in...

    Akinori Andoet al. Transformation of an oleaginous zygomycete Mortierella alpina 1S-4 wit...

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