A selective PCR-based method for the identification of Phytophthora hibernalis Carne

Abstract

Preharvest and postharvest brown rot of citrus fruit is responsible for important economic losses thoroughout the world. The disease is commonly caused by several species of Phytophthora. In Spain, citrus brown rot is mainly caused by P. citrophthora, but in the last years outbreaks of the disease in lemon and sweet orange caused by P. hibernalis Carne have been recorded in northwestern citrus-growing areas. In this work, a PCR method has been developed for the diagnostic of P hibernalis either from isolated mycelia or directly from fruit lesions. One specific primer pair, PHIB1 and PHIB2, was designed from the nucleotide sequences of ITS 1 and ITS2 regions. The two primers amplified a 407-bp fragment from the genomic DNA of P. hibernalis that was sequenced (Ace. No. AY827556). Results demonstrated that PCR amplification of ITS regions by primers PHIB1 and PHIB2 followed by DNA sequencing can provide a rapid, selective and reliable identification of P hibernalis.