Laiou Angeliki, Mandolini Luca Aconiti, Piredda Roberta, Bellarosa Rosanna, Simeone Marco Cosimo
Department of Agriculture, Forests, Nature and Energy (DAFNE) - Università degli Studi della Tuscia, via S. Camillo de' Lellis, 01100 Viterbo, Italy.
Zookeys. 2013 Dec 30(365):197-213. doi: 10.3897/zookeys.365.5670.
Since the pre-historic era, humans have been using forests as a food, drugs and handcraft reservoir. Today, the use of botanical raw material to produce pharmaceuticals, herbal remedies, teas, spirits, cosmetics, sweets, dietary supplements, special industrial compounds and crude materials constitute an important global resource in terms of healthcare and economy. In recent years, DNA barcoding has been suggested as a useful molecular technique to complement traditional taxonomic expertise for fast species identification and biodiversity inventories. In this study, in situ application of DNA barcodes was tested on a selected group of forest tree species with the aim of contributing to the identification, conservation and trade control of these valuable plant resources. The "core barcode" for land plants (rbcL, matK, and trnH-psbA) was tested on 68 tree specimens (24 taxa). Universality of the method, ease of data retrieval and correct species assignment using sequence character states, presence of DNA barcoding gaps and GenBank discrimination assessment were evaluated. The markers showed different prospects of reliable applicability. RbcL and trnH-psbA displayed 100% amplification and sequencing success, while matK did not amplify in some plant groups. The majority of species had a single haplotype. The trnH-psbA region showed the highest genetic variability, but in most cases the high intraspecific sequence divergence revealed the absence of a clear DNA barcoding gap. We also faced an important limitation because the taxonomic coverage of the public reference database is incomplete. Overall, species identification success was 66.7%. This work illustrates current limitations in the applicability of DNA barcoding to taxonomic forest surveys. These difficulties urge for an improvement of technical protocols and an increase of the number of sequences and taxa in public databases.
自史前时代以来,人类就一直将森林作为食物、药物和手工艺品的宝库。如今,利用植物原材料生产药品、草药、茶、烈酒、化妆品、糖果、膳食补充剂、特殊工业化合物和原材料,在医疗保健和经济方面构成了一项重要的全球资源。近年来,DNA条形码技术被认为是一种有用的分子技术,可补充传统分类学专业知识,用于快速物种鉴定和生物多样性清查。在本研究中,对一组选定的林木物种进行了DNA条形码的原位应用测试,目的是为这些珍贵植物资源的鉴定、保护和贸易控制做出贡献。对陆地植物的“核心条形码”(rbcL、matK和trnH-psbA)在68个树木标本(24个分类单元)上进行了测试。评估了该方法的通用性、数据检索的便利性以及使用序列特征状态进行正确物种分配的情况、DNA条形码间隙的存在以及GenBank鉴别评估。这些标记显示出不同的可靠适用性前景。RbcL和trnH-psbA显示出100%的扩增和测序成功率,而matK在一些植物类群中未扩增。大多数物种具有单一单倍型。trnH-psbA区域显示出最高的遗传变异性,但在大多数情况下,种内序列高度分歧表明不存在明显的DNA条形码间隙。我们还面临一个重要限制,因为公共参考数据库的分类学覆盖不完整。总体而言,物种鉴定成功率为66.7%。这项工作说明了DNA条形码技术在分类学森林调查应用中的当前局限性。这些困难促使技术方案得到改进,并增加公共数据库中的序列和分类单元数量。