Abstract: In order to realize the early detection of Pestalotiopsis funerea in Pine needles,the specific primer AF(R/F) was designed,and PCR detection system was established according to the conserved sequence of ITS region of P.funerea.The results showed that the method could amplify a single band of 480 bp in genomic DNA of P.funerea and pine needle genomic DNA. P.funerea could be detected in tissues without obvious symptoms.The established PCR detection system was suitable for molecular identification of P.funerea and early diagnosis of pine needle blight.