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For our specific grayling population under study, we found that the embryos were unable to successfully develop at temperatures above 12°C - indicating a narrow thermal tolerance.
In addition, both common-garden rearing experiments and long-term field observations of spawning temperatures indicate that grayling embryos cannot survive below 5°C, further illustrating the narrow range of temperatures in which embryos of this grayling population can survive.
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This could be important for decisions in conservation and management of grayling populations.
Current management of grayling populations includes habitat restoration and supplementary stocking, but thorough assessment of within-river population structure is clearly needed.
The main goal of the present study was to investigate the genetic diversity of grayling populations in Serbia, using two mtDNA loci (CR and ATP6), in order to clarify the phylogeography of grayling populations in this previously unstudied part of its native range.
Our study shows that (1) the Ibar, Lim and Drina Rivers grayling populations are genetically distinct from populations outside of Serbia and thus should be managed as native populations in spite of some introgression in the Drina River population and (2) the Rzav River population is not appropriate for further stocking activities since it originates from stocked Slovenian grayling.
Analyzing the ATP6 gene in grayling populations of Serbia confirms the division, previously observed from CR sequencing results, into two subclades (the northern i.e. Slovenian and the southern i.e.Serbian) in the Balkan clade defined by the synapomorphy at position 34 of the ATP6 gene (see additional file 2).
The last (Würm) glaciation in Europe ended ~10 000 years ago coinciding with both colonization of the present grayling habitat and decline of grayling populations.
Serbian grayling populations are genetically distinct from Slovenian and other European populations.
Grayling populations elsewhere in Europe clearly are able to develop successfully at these temperatures [ 41].
An even more recent decline of European grayling populations throughout central Europe, due to pollution, habitat destruction, river engineering, predation from piscivorous birds and overfishing [ 1, 13, 15, 25- 27], is also characteristic to Serbian grayling populations.
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