Mogni Virginia Y, Kahan Mariano A, de Queiroz Luciano Paganucci, Vesprini José L, Ortiz Juan Pablo A, Prado Darién E
Facultad de Ciencias Agrarias, Universidad Nacional de Rosario, Campo Experimental Villarino, S2125ZAA Zavalla, Santa Fe Argentina ; IICAR, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET-UNR), Zavalla, Argentina.
SINERGIUM Biotech, Ruta Panamericana Km 38.7, 1619 Garin, Buenos Aires Argentina.
Springerplus. 2016 Apr 18;5:477. doi: 10.1186/s40064-016-2118-4. eCollection 2016.
The Anacardiaceae is an important and worldwide distributed family of ecological and socio-economic relevance. Notwithstanding that, molecular studies in this family are scarce and problematic because of the particularly high concentration of secondary metabolites-i.e. tannins and oleoresins-that are present in almost all tissues of the many members of the group, which complicate the purification and amplification of the DNA. The objective of this work was to improve an available DNA isolation method for Schinopsis spp. and other related Anacardiaceae, as well as the PCR protocols for DNA amplification of the chloroplast trnL-F, rps16 and ndhF and nuclear ITS-ETS fragments. The modifications proposed allowed the extraction of 70-120 µg of non-degraded genomic DNA per gram of dry tissue that resulted useful for PCR amplification. PCR reactions produced the expected fragments that could be directly sequenced. Sequence analyses of amplicons showed similarity with the corresponding Schinopsis accessions available at GenBank. The methodology presented here can be routinely applied for molecular studies of the group aimed to clarify not only aspects on the molecular biology but also the taxonomy and phylogeny of this fascinating group of vascular plants.
漆树科是一个重要且分布于全球的科，具有生态和社会经济意义。尽管如此，由于该科许多成员几乎所有组织中都特别富含次生代谢物，即单宁和油树脂，这使得DNA的纯化和扩增变得复杂，因此对该科的分子研究较少且存在问题。这项工作的目的是改进一种现有的用于Schinopsis属植物及其他相关漆树科植物的DNA提取方法，以及用于叶绿体trnL-F、rps16和ndhF以及核ITS-ETS片段DNA扩增的PCR方案。所提出的改进方法使得每克干燥组织能够提取70 - 120微克未降解的基因组DNA，这些DNA可用于PCR扩增。PCR反应产生了可直接测序的预期片段。扩增子的序列分析显示与GenBank中现有的相应Schinopsis登录号相似。本文介绍的方法可常规应用于该类群的分子研究，旨在不仅阐明分子生物学方面的问题，还能阐明这一迷人的维管植物类群的分类学和系统发育。
