Validation of DNA barcoding as an efficient tool for taxon identification and detection of species diversity in Italian conifers

DNA barcoding may be particularly important in impacting the current trends of world’s biodiversity crisis, public economic issues and science. Forests play a crucial role in self-sustaining natural ecosystems and human environment at both local and regional scales. Over 39% of the world’s forests are formed by conifers; most species are highly exposed to threats and exploited at high frequency. Conifers have an urgent need to be identified through DNA barcodes, but received little attention up to date. The application of standard barcoding protocols appears to be hampered by a lack of consensus on the selection of markers and low rates of successful species identification. We examined whether four marker regions (trnh-psba, rbcL, rpoc1 and matK) matched conifer species taxonomy in a practical biodiversity survey of Italian forested land. Our objective was to provide a test of future in situ applications of DNA barcodes by evaluating the efficacy of species discrimination under the criteria of uniformity of methods and natural co-occurrence of the species. Twenty-five conifer species were included in a floristic study based on regional samplings. Genetic distance, similarity and phylogeny approaches were used to assess identification of taxa and species monophyly. We obtained 100% total discrimination success, with rbcL?+?trnH-psbA as the suggested two-locus barcode in terms of universality and efficacy. Consistent barcoding gaps, supported monophyly and regional patterns of diversity, were evidenced. Species requiring further investigation to be fully assessed at a broader context are indicated. We conclude that the time seems now ripe for coherent barcoding campaigns of entire forest tree regional floras.