Results from metabarcoding communities from the National Parks published in PeerJ Preprints

by Xavier Turon

While the article is being peer-reviewed, we have published the manuscript in Pre-print format in the repository PeerJ Preprints, where it is freely available at Wangensteen OS, Palacín C, Guardiola M, Turon X. 2017. Metabarcoding litoral hard-bottom communities: unexpected diversity and database gaps revealed by two molecular markers. PeerJ Preprints 5:e3429v1.

We report in this manuscript the astonishing diversity found in communities from Islas Cies and Cabrera. We address several technical aspects, compare the two genes studied (18S and COI), and perform ecological analyses. Here goes the abstract:

We developed a metabarcoding method for biodiversity characterization of structurally complex natural marine hard-bottom communities. Novel primer sets for two different molecular markers: the “Leray fragment” of mitochondrial cytochrome c oxidase, COI, and the V7 region of ribosomal RNA 18S were used to analyse eight different marine shallow benthic communities from two National Parks in Spain (one in the Atlantic Ocean and another in the Mediterranean Sea). Samples were sieved into three size fractions from where DNA was extracted separately. Bayesian clustering was used for delimiting molecular operational taxonomic units (MOTUs) and custom reference databases were constructed for taxonomic assignment. We found unexpectedly high values for MOTU richness, suggesting that these communities host a large amount of yet undescribed eukaryotic biodiversity. Significant gaps are still found in sequence reference databases, which currently prevent the complete taxonomic assignation of the detected sequences. Nevertheless, over 90% (in abundance) of the sequenced reads could be successfully assigned to phylum or lower taxonomical level. This identification rate might be significantly improved in the future, as reference databases are updated. Our results show that marine metabarcoding, currently applied mostly to plankton or sediments, can be adapted to structurally complex hard bottom samples, and emerges as a robust, fast, objective and affordable method for comprehensively characterizing the diversity of marine benthic communities dominated by macroscopic seaweeds and colonial or modular sessile metazoans, allowing for standardized biomonitoring of these ecologically important communities. The new universal primers for COI can potentially be used for biodiversity assessment with high taxonomic resolution in a wide array of marine, terrestrial or freshwater eukaryotic communities.

Percent of MOTUs found for the main categories of organisms in the three fractions studied with 18S (left) and COI (right) in all communities.

The methods developed for metabarcoding of hard bottom communities explained in an international book chapter

by Xavier Turon

Owen Wangensteen and Xavier Turon have been invited to write a chapter in the book "Marine Animal Forests. The Ecology of Benthic Biodiversity Hotspots", of the Springer International Publishing Company (ISBN: 978-3-319-17001-5), which is edited by Sergio Rossi, Lorenzo Bramanti, Andrea Gori and Covadonga Orejas. In this chapter, entitled "Metabarcoding techniques for assessing biodiversity of marine animal forests", Owen and Xavier explain the methods developed for analysing hard bottom communities using metabarcoding. We believe that it is crucial to setup standardized protocols, and that this book can be an excellent platform to share and disseminate our experience with the different steps of the metabarcoding process.

This chapter is now accepted for publication, and we foresee that it will be published in the forthcoming months.

Techniques developed in METABARPARK, such as the size fractionation of the samples, can be adopted by other teams working on metabarcoding of marine benthos.

Measuring settlement rates of sea urchins. New paper out!

by Owen Wangensteen

Team members of the ChallenGen Project at the University of Barcelona, in collaboration with scientists from the University of La Laguna (Canary Islands) have just published a new paper on Scientia Marina journal, describing and testing new designs of artificial collectors for quantitative assessment of settlement rates in sea urchins.

The study of population dynamics of sea urchins is crucial for understanding the ecology of Mediterranean and temperate Atlantic rocky shores, since these echinoderms are often the main herbivore species in shallow benthic ecosystems and they are responsible of linking the primary production from seaweeds to other higher levels of the trophic chains. Many sea urchin populations may be limited by the settlement rates (the process by which the planktonic sea urchin larvae settle and become juvenile sea urchins). Methods for measuring these settlement rates were not standardized and yielded non-comparable results. The work presented in this paper, carried out in parallel at Canary Islands and Costa Brava (Spain) shows that three-dimensional collectors made from “bioballs” (a biofilm-friendly, high surface material often used as filters in aquaria) are the most suitable device for this assessment, allowing for an accurate and repeatable measurement of this elusive, yet important, ecological process.

The best part is that these bioballs collectors are useful not only for sea urchins settlers. They could be suitable for assessing settlement and colonization rates for a wide-range of marine invertebrates and can help marine biologists to get a more accurate view of the population dynamics of many species present in benthic ecosystems.

The three types of collector tested in this work: plastic biofilter ball (A, C), vertical brush (B, D) and horizontal triangular mat of coconut fibre (B, E).