Download Towards estimating invasion potential: Cellular Stress - DNAqua-Net

First attempt to create DNA barcode reference library for
freshwater gammarids of the Periadriatic Region
Kamil Hupało, Tomasz Mamos, Weronika Wrzesińska, Tomasz Rewicz,
Anna Wysocka, Remi Wattier, Michał Grabowski
Our projects focusing on Periadriatic Region
• Neogene and Quaternary sea level changes and origin of epigean
freshwater malacostracan fauna in the periadriatic region 2015-2018
• Cryptic diversity and phylogeography of amphipods from “Gammarus
roeselii” and “G. balcanicus” groups in Europe 2010-2014
• Diversity and origin of the freshwater fauna of the Mediterranean Islands
on the example of Gammaridae (Amphipoda) 2016-2019
All funded by the National Committee on Scientific Research
Snapshot history of the Mediterranean
200 - 20 mln years ago (early Miocene)
Tethys Ocean, Paratethys
Snapshot history of the Mediterranean
12 - 8 mln years ago (Tortonian)
appearence of the proto-Mediterranean Sea
Snapshot history of the Mediterranean
5.96 – 5.33 mln years ago (Messinian)
Messinian Salinity Crysis (MSC), Lago Mare episode
Snapshot history of the Mediterranean
2.5 – 0.016 mln years ago (Pleistocene)
sea regressions, Mega-Po River basin
Material
Amphipoda - Gammaridae
Aims
• Estimating the biodiversity of Periadriatic freshwater gammarids
• Creating a DNA barcode reference library for the Periadriatic
Region
• Revealing biogeographical affiliations of freshwater gammarids
(Amphipoda, Gammaridae) using molecular markers
• Providing spatiotemporal framework for the species origin and
diversification by reconstructing their time-calibrated phylogeny
Sampling sites
Materials & methods
• I step: Morphological analyses
 according to the taxonomical keys
 using SEM
• II step: DNA Barcoding
 COI marker
 BOLD upload
 MOTU delimitation (BIN, ABGD, GMYC, bPTP)
• III step: additional amplification of other markers
 mitochondrial – 16S rRNA
 nuclear – 28S rRNA, ITS, EF-α
• IV step: Data analysis
 evolutionary models, molecular clock,
phylogeny reconstruction etc.
Results of our studies
Gammarus balcanicus group
• 50 sampling
sites
• 318 individuals
barcoded
• 40-45 MOTUs
Gammarus balcanicus group
Gammarus roeselii group
• 26 sampling sites
(23 in Periadriatic)
• 177 individuals
barcoded
• 12-28 MOTUs
Gammarus roeselii group
Gammarus pulex group
Italy and Greece
• 55 sampling sites
• 219 individuals
barcoded
• 19-29 MOTUs
Gammarus pulex group
Italy
Gammarus pulex group
Greece
Gammarus pulex group
Italy & Greece
Echinogammarus
Italy and Greece
• 42 sampling sites
• 225 individuals
barcoded
• 29-30 MOTUs
Echinogammarus
Italy
Echinogammarus
Greece
Echinogammarus
Italy & Greece
Summary
• Material collected from over 500 sampling sites
• So far over 900 barcoded individuals from 170 sampling sites
• High cryptic diversity in freshwater gammarids from Periadriatic:
– More than 40 MOTUs of G.balcanicus group
– At least 12 MOTUs of G.roeselii group
– Nearly 20 MOTUs of G.pulex group
– Over 30 MOTUs of Echinogammarus
Significance
• Freshwater biodiversity assessment of the Periadriatic region
• Creating DNA barcode reference library for future studies
• Planning reasonable and effective strategy preventing the loss
of biodiversity
• Understanding evolutionary mechanisms and historical factors
behind formation of freshwater fauna