China Agricultural University scientists developed a gene disruption method for plant pathogen Botryosphaeria dothidea using polyethylene glycol-mediated protoplast transformation. The details of the method are published in BMC Biotechnology.
Botryosphaeria dothidea causes apple white rot and infects several tree plants. Though the genome data for this pathogen are available and many pathogenesis-related genes have been predicted, there is a need for a gene manipulation method to understand the pathogenic mechanism of B. dothidea.
Using the developed method, the gene homologous recombination cassette resulted in higher gene disruption than a GHR plasmid. A high GD efficiency and low frequency of random insertions were observed when using the developed method. Furthermore, disruption of genes was attained in two strains. One strain produced less melanin, while the other one exhibited a slower growth rate and a stronger resistance to Congo red test.