AFLP Analysis of Single-Zoospore Isolates of Phytophthora ramorum
Christopher Fung1, Kelly Ivors2, Matteo Garbelotto2
Phytophthora ramorum, the oomycete pathogen responsible for Sudden Oak Death, is thought to reproduce asexually through production of zoospores, chlamydospores and sporangia. AFLP (Amplified Fragment Length Polymorphism) analyses were conducted on ten sets of ten single-zoospore isolates. These samples were generated from cultures of P. ramorum collected from five different host species (Quercus agrifolia, Quercus kelloggii, Umbellularia californica, Rhododendron spp., and Lithocarpus densiflora) located in different Northern California counties (Marin, Sonoma, Solano, Santa Cruz and San Mateo). Genomic DNA was extracted from pure cultures of each single zoospore isolate and digested with the restriction enzymes EcoRI and MseI. Fragments of the genomic DNA were ligated to standard blunt end adaptors and amplified by two successive rounds of PCR (Polymerase Chain Reaction). This resulted in the creation of a unique genetic "fingerprint". Fingerprint patterns were analyzed using an ABI 3100 Genetic Analyzer. Genetic similarity or variation between single-zoospore isolates determined from these analyses will be presented.
1University of California, Department of ESPM-ES, 23 Hilgard Hall, Berkeley, CA 94720; (510) 643-4282; cfung@nature.berkeley.edu
2University of California, Department of ESPM-ES, 23 Hilgard Hall, Berkeley, CA 94720
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