IDENTIFYING PINE-INHABITING LOPHODERMIUM SPECIES USING PCR-RFLP*

Three species of Lophodermium are reported from pine in New Zealand—the previously recorded L. conigenum (Brunaud) Hilitzer and L. pinastri (Schrad.) Chevall., and the newly reported L. molitoris Minter. All are saprobes with an initial endophytic phase. The North American species L. seditiosum Minter, Staley & Millar is considered a potential threat to New Zealand’s plantation forests. Because these fungi are difficult to distinguish morphologically, a simple molecular method was developed to distinguish the four species. After PCR amplification, the ITS * region is digested using HaeIII and HpaII in a single reaction, resulting in unique RFLP banding patterns for each of the species treated. A phylogenetic analysis based on ITS sequences revealed two genetically distinct groups within L. pinastri, one North American and one European. All the New Zealand isolates of L. pinastri tested were identical to the North American group. Further development of the method described here, so that fungal DNA is targeted directly from infected plant tissue, will provide an effective quarantine tool for New Zealand’s forestry industry