Spizellomyces punctatus
Spizellomyces punctatus is a chytrid fungus living in soil. It is a saprotrophic fungus that colonizes decaying plant material. Being an early diverging fungus, S. punctatus retains ancestral cellular features that are also found in animals and amoebae. Its pathogenic relatives, Batrachochytrium dendrobatidis and B. salamandrivorans, infect amphibians and cause global biodiversity loss. The pure culture of S. punctatus was first obtained by Koch (named Phlyctochytrium punctatum). Genome The genome of S. punctatus strain DAOM BR117 was sequenced under the Origins of Multicellularity project. Its genome size is about 24.13 Mb with a GC content of 47.6%. The genome has 9,424 predicted transcripts and 8,952 predicted protein-coding genes. The DDBJ/EMBL/GenBank accession number is ACOE00000000. Genetic transformation Agrobacterium-Mediated Transformation Genetic transformation of S. punctatus zoospores by plant pathogen Agrobacterium tumefaciens EHA105 strain is successfully established. Several selection markers have been tested. The growth of S. punctatus is not inhibited by Geneticin (G418), Puromycin, and Phleomycin D10 (Zeocin) up to 800 mg/L. 200 mg/L Hygromycin and 800 mg/L Nourseothricin (CloNAT) completely inhibit S. punctatus growth. The scientists who develop this protocol use Hygromycin as the selection marker. S. punctatus HSP70 and H2B promoters drive sufficient gene expression for Hygromycin resistance and GFP