Abstract: DNA was extracted from the endosperm of five needles pine by improved CTAB method. The purity of DNA that extracted by this method was high, which was tested by UV spectrophotometer. The yield of single seed's endosperm DNA almost could meet the need of mass PCR amplification. The best stable reduplicative ISSR-PCR reaction systems of five needles pine and PCR amplification parameters were established. Sixteen ISSR-PCR primers that had vivid amplification band and abundant polymorphism had been screened. The work provides a standard program for using need's endosperm of pine to carry out establishment of genetic fingerprinting and other studies of molecular biology in the future.

Key words: five needles pine, seed's endosperm, DNA extracted, ISSR-PCR