Agricultural Biotechnology Research Institute for Northwest & West of Iran, Tabriz, Iran.
Mol Biol Rep. 2010 Feb;37(2):833-7. doi: 10.1007/s11033-009-9634-z. Epub 2009 Aug 2.
Various investigations have been so far performed for extraction of genomic DNA from plant tissues, in which the extracted intact DNA can be exploited for a diverse range of biological studies. Extraction of high quality DNA from leathery plant tissues (e.g., coniferous organs) appears to be a critical stage. Moreover, for some species such as Taxus trees, bioprocess engineering and biosynthesis of secondary metabolites (e.g., paclitaxel) is a crucial step due to the restrictions associated with extinction of these species. However, extraction of intact genomic DNA from these plants still demands a rapid, easy and efficient protocol. To pursue such aim, in the current work, we report on the development of a simple and highly efficient method for the extraction of DNA from Taxus baccata. Based upon our protocol, interfering phenolic compounds were removed from extraction using polyvinylpyrrolidone and RNA contamination was resolved using LiCl. By employing this method, high quality genomic DNA was successfully extracted from leaves of T. baccata. The quality of extracted DNA was validated by various techniques such as RAPD marker, restriction digestions and pre-AFLP. Upon our findings, we propose this simple method to be considered for extraction of DNA from leathery plant tissues.
目前已经进行了各种从植物组织中提取基因组 DNA 的研究，从中提取的完整 DNA 可用于广泛的生物学研究。从革质植物组织（例如针叶器官）中提取高质量的 DNA 似乎是一个关键阶段。此外，对于某些物种，如紫杉树，由于与这些物种灭绝相关的限制，生物加工工程和次生代谢物（例如紫杉醇）的生物合成是一个关键步骤。然而，从这些植物中提取完整的基因组 DNA 仍然需要一种快速、简单和有效的方法。为了实现这一目标，在目前的工作中，我们报告了一种从欧洲红豆杉中提取 DNA 的简单而高效的方法。根据我们的方案，通过使用聚乙烯吡咯烷酮去除提取过程中的干扰性酚类化合物，并使用 LiCl 解决 RNA 污染。通过采用这种方法，成功地从欧洲红豆杉的叶片中提取出高质量的基因组 DNA。通过 RAPD 标记、限制酶消化和预 AFLP 等各种技术验证了提取 DNA 的质量。根据我们的发现，我们建议将这种简单的方法用于从革质植物组织中提取 DNA。
