Abstract
PCR products were obtained from the apple pathotype of Alternaria alternata with two sets of primers whose design was based on the aligned amino acid activating domains of cyclic peptide synthetase genes. Some of these products were subcloned and sequenced and all sequences showed considerable homology to the corresponding region of other CPSs. These were used as probes against genomic digests of saprophytic and other A. alternata pathotypes. Hybridization at low stringency produced several signals of varying size from all the strains examined, but only A. alternata apple pathotype hybridized at high stringency. Two of the sequences were then subcloned into a vector and used for gene targeting. Three out of seventy transformants appeared to lack AM-toxin production, based on both a leaf necrosis assay and HPLC analysis. However, analysis of these transformants showed that the subcloned sequence hybridized to a fragment that was the same size in the wild type, although the signal intensity was significantly lower. This may indicate a possible multi-copy nature for the AM-toxin synthetase gene. The integration profile of the transforming vector in these transformants has not been solved, so the loss of AM-toxin production is not conclusively due to a gene disruption event.
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Itoh, Y. et al. (1998). A Catalytic Domain of a Cyclic Peptide Synthetase that is Specific for the Apple Pathotype of Alternaria Alternata and its Possible Involvement in Host-Specific AM-Toxin Production. In: Kohmoto, K., Yoder, O.C. (eds) Molecular Genetics of Host-Specific Toxins in Plant Disease. Developments in Plant Pathology, vol 13. Springer, Dordrecht. https://doi.org/10.1007/978-94-011-5218-1_6
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