Molecular Analysis of an Endopolygalacturonase Gene from a Eucalyptus Canker Pathogen, Cryphonectria cubensis

Abstract

Cryphonectria cubensis causes a serious Eucalyptus canker disease. Fungal cell wall degrading enzymes (CWDEs) are important during the early stages of interaction of the fungus with Eucalyptus. To improve our understanding of the molecular regulation of the interaction of Eucalyptus and C. cubensis, the relevant genes involved in this interaction should be identified, cloned and studied. The aim of this study was, therefore, to clone the endopolygalacturonase (endoPG) gene of C. cubensis. C. cubensis was grown on a medium supplemented with Eucalyptus cell wall extracts. Degenerate primers were designed to amplify part of the endoPG gene from C. cubensis genomic DNA. The resulting sequence was used to design specific primers for use in inverse PCR to amplify the entire endoPG gene of C. cubensis (ccen-1). The endoPG sequence of C. cubensis has 93% amino acid sequence similarity to that of the chestnut blight pathogen, Cryphonectria parasitica.