CLONING OF 2 CHITIN SYNTHASE GENE FRAGMENTS FROM A PROTOPLASTIC ENTOMOPHTHORALE

Chitin synthase expression was studied in spontaneously produced proto plasts and in hyphal bodies of the Entomophthorale species Entomophaga aulicae. The absence of wall in protoplasts was correlated to an abse nce of chitin synthase. Two chitin synthase activities with different biochemical characteristics have been detected in the hyphal bodies. T wo chitin synthase gene fragments EaCHS1 and EaCHS2 of 600 bp were obt ained using PCR amplification of genomic DNA. Their amino acid sequenc es showed 75% identity. Compared with other fungal chitin synthases, t hey belong to class II. EaCHS1 and EaCHS2 were used to probe total RNA from E. aulicae hyphal bodies and protoplasts. A single transcript of 2.4 kb hybridized only with EaCHS1 in protoplasts and hyphal bodies.