Towards the establishment of a DNA-barcoding iden8fica8on system for Iberian plants Ricarda Riina Real Jardín Botánico (CSIC, Madrid),
[email protected]
Introduc*on The ini8al establishment of a DNA-barcoding system depends heavily on taxonomists because of the need of correctly iden8fied specimens associated to the selected barcodes (Meyer and Paulay 2005; Galimber8 2015b). Once established, the system speeds up the process of species iden8fica8on and biodiversity discovery for which trained taxonomists are progressively scarcer or are limited in the 8me they can dedicate to iden8fica8on tasks.
DNA-barcoding ini8a8ves have proved the opposite effect and are seen as invigora8ng and complementary for taxonomic research (Newmaster 2008; Packer et al. 2009; Zhang et al. 2015)
Here we present a new ini8a8ve to establish a DNA-barcoding system for Iberian plants: the IbeBoL project. The Iberian flora, with about 54% of the European flora of vascular plants (Aedo 2013), represents one of the most species-rich aeas of the con8nent. It harbours ca. 6273 taxa, including many groups with unresolved taxonomy.
Objec*ves •
Establish protocols, quality criteria and standards for a full DNA-barcoding ini8a8ve, and the ini8a8on of a DNA bank for the Iberian flora.
•
Evaluate the poten8al of RJB (Real Jardín Botánico) collec8ons and in-house resources with the intend to lower the costs of building a barcode library of Iberian plants.
•
Carry out a pilot test of a group of DNA barcodes on a selec8on of divergent groups of Iberian plants.
•
Explore the use of High-throughput sequencing technologies (e.g., PacBio) as an alterna8ve to Sanger sequencing.
Taxon sampling strategy Approximately 250 taxa occurring in the Iberian Península will be targeted, each represented by 1-6 specimens depending on distribu8on range and samples availability. Rare species will probably be represented by fewer samples (1-2), and widespread ones will have up to 5-6 samples depending on size of geographic range. Having several individuals from different and separate locali8es is important to evaluate barcode intraspecific varia8on.
Following this strategy, we will have about 1000 samples (DNA isolates) from the selected 250 taxa, that will be tested for the four proposed barcode markers (see below).
Taxa will be selected to represent divergent clades from the vascular plant tree of life.
For this pilote phase, each of the genera/groups should meet the following criteria:
• The group is well represented in the Iberian Peninsula • It has been treated in a taxonomic study (e.g. Flora Iberica or elsewhere) or has a taxonomist at the Real Jardín Botánico (RJB) or at a partner ins8tu8on. • It has a significant number of collec8ons at RJB, especially in the herbarium (MA).
Collabora*ons
Barcodes and sequencing approaches
Our project (IbeBoL) has ini8ated collabora8ons with two research teams with ongoing projects aiming to develop DNA-barcoding iden8fica8on systems for their respec8ve na8onal and regional floras. One is the German Barcode of Life ini8a8ve (GBoL) (Geiger et al. 2016) and the other is the Canary Islands plant DNA-Barcoding group based at the Jardín Botánico Canario “Viera y Clavijo” (Jaén-Molina et al. 2014). Besides exchanging crucial informa8on on laboratory protocols, methods, sample storage sesngs and data processing, we will also try to test and eventually adopt some of their bioinforma8cs tools.
For the ini8al phase of the IbeBoL project we will work with the same set of markers used by the botany sec8on of the German Barcode of Life ini8a8ve (GBoL). These barcodes are three fastevolving plas8d DNA regions (trnL-F, rpl16, trnK/matK) in combina8on with the internal transcribed spacers (ITS) of the nuclear ribosomal genes. Although the original IbeBoL project was conceived to use a Sanger sequencing approach we will try to explore High-throughput sequencing technologies, which are already being implemented by our GBoL partners.
Acknowledgments We are grateful to Quandt (Nees Ins8tute for Plant Biodiversity; GBoL botany sec8on), Ruth Jaén-Molina and Juli Caujapé-Castells (Jardín Botánico Canario “Viera y Clavijo-Unidad Asociada CSIC) for their support and collabora8on. The IbeBoL project (CGL2015-73621-JIN) is funded by the Agencia Estatal de Inves8gación (AEI) and the European Regional Development Fund (FEDER).
References Aedo, C. et al. 2013. Species richness and endemicity in the Spanish vascular flora. Nordic Journal of Botany 31:478-488. Geiger, M.F. et al. 2016. How to tackle the molecular species inventory for an industrialized na8on—lessons from the first phase of the German Barcode of Life ini8a8ve GBOL (2012–2015). Genome 59:661-670. Galimber8, A. et al. 2015. DNA barcoding in mammals: what’s new and where next? Hystrix, the Italian Journal of Mammalogy 26: 13-24. Jaén-Molina, R., et al. 2015. Molecular taxonomic iden8fica8on in the absence of a ‘barcoding gap’: a test with the endemic flora of the Canarian oceanic hotspot. Molecular ecology resources 15: 42-56. Meyer, C.P. & Paulay, G. 2005. DNA Barcoding: Error Rates Based on Comprehensive Sampling. PLOS Biol. 3: e422. Newmaster, S.G., et al. 2008. Tes8ng candidate plant barcode regions in the Myris8caceae. Molecular ecology resources 8: 480-490. Packer, L. et al. 2009. The status of taxonomy in Canada and the impact of DNA barcoding. Canadian Journal of Zoology 87: 1097-1110. Zhang, J.Q. et al. 2015. DNA Barcoding of Rhodiola (Crassulaceae): A Case Study on a Group of Recently Diversified Medicinal Plants from the Qinghai-Tibetan Plateau. PloSone 10: e0119921.
