Drábková Lenka Záveská
Department of Taxonomy, Institute of Botany, Academy of Sciences of the Czech Republic, Pruhonice, Czech Republic.
Methods Mol Biol. 2014;1115:69-84. doi: 10.1007/978-1-62703-767-9_4.
With the expansion of molecular techniques, the historical collections have become widely used. Studying plant DNA using modern molecular techniques such as DNA sequencing plays an important role in understanding evolutionary relationships, identification through DNA barcoding, conservation status, and many other aspects of plant biology. Enormous herbarium collections are an important source of material especially for specimens from areas difficult to access or from taxa that are now extinct. The ability to utilize these specimens greatly enhances the research. However, the process of extracting DNA from herbarium specimens is often fraught with difficulty related to such variables as plant chemistry, drying method of the specimen, and chemical treatment of the specimen. Although many methods have been developed for extraction of DNA from herbarium specimens, the most frequently used are modified CTAB and DNeasy Plant Mini Kit protocols. Nine selected protocols in this chapter have been successfully used for high-quality DNA extraction from different kinds of plant herbarium tissues. These methods differ primarily with respect to their requirements for input material (from algae to vascular plants), type of the plant tissue (leaves with incrustations, sclerenchyma strands, mucilaginous tissues, needles, seeds), and further possible applications (PCR-based methods or microsatellites, AFLP).
随着分子技术的扩展，历史标本收藏已得到广泛应用。利用DNA测序等现代分子技术研究植物DNA，在理解进化关系、通过DNA条形码进行鉴定、保护状况以及植物生物学的许多其他方面发挥着重要作用。大量的植物标本馆收藏是重要的材料来源，特别是对于来自难以进入地区或现已灭绝类群的标本。利用这些标本的能力极大地促进了研究。然而，从植物标本中提取DNA的过程常常充满困难，这些困难与植物化学、标本干燥方法以及标本化学处理等变量有关。尽管已经开发出许多从植物标本中提取DNA的方法，但最常用的是改良的CTAB和植物DNeasy Mini试剂盒方案。本章中选择的九种方案已成功用于从不同种类的植物标本馆组织中提取高质量的DNA。这些方法的主要区别在于对输入材料（从藻类到维管植物）的要求、植物组织的类型（有结壳的叶子、厚壁组织束、粘液组织、针叶、种子）以及进一步可能的应用（基于PCR的方法或微卫星、AFLP）。
