783 SEAKEYS: unlocking foundational marine biodiversity knowledge in South Africa using DNA barcoding

1,2Monica Mwale, 2G. Gouws, 3J Landschoff, 3JA Toms, 3CL Griffiths and 4K. Sink

1National Zoological Gardens of South Africa, PO BOX 754, Pretoria 0001, South Africa. 2South African Institute for Aquatic Biodiversity, P/Bag 105, Grahamstown, South Africa 3Zoology Department, Marine Biology Research Centre, University of Cape Town, Rondebosch, South Africa 4South African National Biodiversity Institute, Kirstenbosch Research Centre, Cape Town, South Africa

ABSTRACT The information on the biodiversity of most southern African marine ecosystems is limited1. The development of comprehensive databases is therefore critical to support the effective monitoring of marine ecosystems and species for the implementation of evidence-based policy and conservation management for sustainable use. The SEAKEYS project funded by the DST/NRF/SANBI Foundational Biodiversity Knowledge and Information research funding Programme (FBIP) therefore aims to unlock fundamental biodiversity information using DNA barcoding to generate species records and distribution databases for priority ecosystems and groups.

AIMS & OBJECTIVES High levels of genetic variation

To describe and undertake associated genetic (DNA barcoding),  Taxonomically discordant BINS (28) with good sample sizes (5-65 systematic and ecological research for; samples) were re-analysed and corrected using the DNA barcode.

 Priority taxa: harvested, habitat forming, indicator, threatened and Example: Genetic variation of Gymnothorax undulatus potentially invasive species. (Anguilliformes: Muraenidae) in the WIO

 Priority ecosystems: with key ecosystem services and those that are G. undulatus is an Indo-Pacific species defined by presence of particularly sensitive to fishing, mining and other impacts. irregular brown rectangular spots with reticulated pattern1,2,4

The research aims to support biodiversity assessments and evidence-  Taxonomic status is unclear and the species is highly misidentified based decision making and effective marine biodiversity. and confused with other Indo-Pacific species3,4.

 This was observed in both DNA barcode data and morphological RESULTS analysis.

 DNA barcodes of 573 specimens of coastal marine fishes and

invertebrates from 128 nominal species GG07-A080 G. undulatus SA HM09-091 Gymnothorax sp. SA GG07-A079 G. undulatus SA GG07-A088 G. undulatus SA Clade 2 99 GG07-A089 G. undulatus SA Clade 2  Collected from the cool-temperate, warm-temperate and sub-tropical 100 HM09-064 G. undulatus SA Smith-41.24 1 G. undulatus SA HM09-065 G. undulatus SA T-355 G. griseus bioregions and offshore (Agulhas Bank) in South Africa, mascarene T-453 G. chilospilus KNY2011/164 Gymnothorax sp. EA RB09-144 Gymnothorax sp. Moz plateau and Inhaca Island in southern Mozambique. RB09-104 Gymnothorax sp. Moz 100 DB11-103 G. undulatus SA 99 T-341 G. rueppellii T-340 G. rueppellii MAL226 G. undulatus Arc 85  These specimens included invasive, commercially important, rare, 100 LS07-0793 G. undulatus EA LS07-0638 G. undulatus EA Clade 1 100 T-346 G. undulatus Sey Clade 2 threatened and endangered species. Average COI sequence T-421 G. undulatus Sey 87 T-422 G. undulatus Sey CB10-351 Gymnothorax sp. EA divergences within species, genera and families were moderate to high GG07-A050 G. flavimarginatus 98 RB10-A014 G. flavimarginatus JX242970 G. flavimarginatus at 1.76%, 20.0% and 23.5% respectively. 100 JX242970 G. flavimarginatus AB695709 G. undulatus Jap AB695707 G. undulatus Jap Clade 3 100 AB695708 G. undulatus Jap Clade 3 AB695653 G. thyrsoideus 100 AB695651 G. thyrsoideus  The barcode-gap analysis indicated that 14 species were indistinct, AB695652 G. thyrsoideus 88 JX242971 G. fuscomaculatus JX242968 G. chilospilus while 23 had high intraspecific distances (>2%) due to either deep 100 AB695696 G. chilospilus 100 AB695695 G. chilospilus 95 AB695694 G. chilospilus divergences or cryptic speciation. AB695649 G. pictus AB695650 G. pictus 100 AB695648 G. pictus JX242976 G. pictus AB702979 S. tigrina AB702972 S. tigrina 100 AB702973 S. tigrina  An additional 12 species had very low interspecific variation suggesting 92 AB702976 S. tigrina misidentifications or a need for taxonomic evaluation. 0.02 Figure 2: DNA barcoding (COI) ML tree and WIO colour  The 138 Barcode Index Numbers (BIN) identified included 93 morphs of G. undulatus in the WIO (Top: South Africa, Middle: concordant assignments. Seychelles, Bottom: FISHBASE distribution map.  17 singletons specimens that will require more sampling for species verification. SIGNIFICANCE

2% 1% Southern African marine taxa are very Arthropoda (phylum) diverse, with more species to be discovered. 4% Mollusca (phylum) 6% Cnidaria (phylum) BINs are higher than the number of 7% 37% identified species. Echinodermata (phylum) 8% Chordata (phylum) The vouchered specimens from this study will Annelida (phylum) be critical in linking all new and unclear BIN clusters to species and new descriptions. Bryozoa (phylum) 35% 4 others (phylum) Thus more holotype and type locality barcodes will become vital.

Barcodes consistent with good morphological identifications for species.

Therefore, this project will contribute towards the estimation of marine biodiversity in South Africa.

Funding Sources/Acknowledgements

1. DST/NRF/SANBI Foundational Biodiversity Knowledge Figure 1: Summary of taxa analysed from SeaKeys project. 2. Dr Kerry Sink – SANBI Graph show diversity among fishes (e.g. endemic species of 3. Yonela Sithole (MSc) 4. BOLD team Cafforgobius). 5. IZIKO museums

References 1. Branch GM, Griffiths CL, Branch ML, Beckley LE. 1994. Two Oceans: A Guide to the Marine Life of Southern Africa. David Phillip Publishers (Pty) Ltd, South Africa, pp 1-456 2. Reece JS. 2010. Phylogenetics and phylogeography of moray eels (Muraenidae). PhD thesis, Washing University, St. Louis, pp 1-112. 3. Reece JS, Bowen BW, Smith DG, Larson A. 2010a. Molecular phylogenetics of moray eels (Muraenidae) demonstrates multiple origins of a shell-crushing jaw (Gymnomuraena, Echidna) and multiple colonizations of the Atlantic Ocean. Molecular Phylogenetics and Evolution 57: 829-835. 4. Castle PHJ, McCosker JE. 1986. Family Muraenidae. In: Smith MM, Heemstra PC (eds), Smiths’ Sea Fishes. MacMillan, South Africa, pp 165-176. Email: [email protected]