竹子DNA提取方法改良及ITS-RFLP分析的初步研究

Improvement of Extracting Genomic DNA from Bamboo Species and RFLP Analysis of PCR Amplified rDNA Internal Transcribed Spacer Region

通过对传统方法的比较、改进,形成一种快速、简易的适用于竹子DNA提取的方法。另外,通过PCR扩增和7种限制性内切酶(AluⅠ、BbeⅠ、BbvⅠ、DpnⅠ、EagⅠ、hindⅢ和RsaⅠ)酶切,对云南世博园内的灰香竹、秀叶箭竹、云南箭竹和滑竹等4个竹种rDNA-ITS进行分析,发现竹子种间甚至是属间的rDNA-ITS序列的RFLP图谱差异很小,不适宜用来作竹子种属鉴定的标准。

A better way of extracting genomic DNA from bamboos was found via comparing with and improving the traditional ways. The rDNA internal transcribed spacer （ITS） regions of Chimonocalamus pollens, Fargesia yuanjiangensis, Fargesia yunnaaensis and Yushania polytricha were amplified with universal primers from the purified genomic DNA. The RFLP analysis was generated with seven restriction endonuclenses such as AluⅠ , Bbe Ⅰ , Bbv Ⅰ, Dpn Ⅰ, EagⅠ , hind Ⅲand Rsa Ⅰ . The results showed that the differences of RFLP fingerprintings of these bamboos were very small, which were unsuitable to identify the bamboo species and genras.