五针松胚乳ISSR-PCR反应体系的建立

采用改良的CTAB法提取5种五针松胚乳DNA，经检测，该方法提取的DNA纯度和含量较高，单粒种子胚乳DNA得率基本满足大量PCR扩增的需要。建立了稳定的、可重复的五针松ISSR-PCR最佳反应体系及PCR扩增程序，筛选出了扩增条带清晰、多态性丰富的16个ISSR引物，为今后利用五针松种子胚乳开展遗传图谱构建等分子生物学研究提供一个标准化程序。

THE ISSR-PCR REACTION SYSTEM'S ESTABLISHMENT ABOUT SEED'S ENDOSPERM OF FIVE NEEDLES PINE

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.