Integrated Method for Sockeye Salmon Stock Differentiation in the West Pacific and the Sea of Okhotsk
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Russian Federal Institute of Fisheries and Oceanography Moscow, Russia INTEGRATED METHOD FOR SOCKEYE SALMON STOCK DIFFERENTIATION IN THE WEST PACIFIC AND THE SEA OF OKHOTSK ANASTASIA KHRUSTALEVA Distribution of sockeye salmon in Asia Chukotka West Coast of the Sea of Okhotsk Kamchatka Bering Sea Peninsula Kuril North Pacific Ocean Islands Sockeye salmon population structure studies Population differentiation methods Biological markers Genetic markers Tagging Morphological Parasitologic Immune-genetic Molecular genetic Biochemical Micro- DNA- mtDNA SNP satellites fingerprinting RFLP AFLP RAPD Micosatellite markers Microsatellite Microsatellite locus repeat Allele 1 DNA Allele 1 molecule Allele 2 Allele 2 Mcrosatellite DNA polymorphism Distribution of sockeye salmon Chukotka peninsula West Coast of the Sea of Okhotsk Kamchatka Bering Sea Peninsula Kuril Islands North Pacific Ocean - Sockeye salmon areal - Unexplored area Populations studied Ch – Chukotka peninsula Kur – North Kuril Islands Okh – Okhota R. Kamchatka peninsula: KPh - Pakhacha R.; KK - Kamchatka R., late run; KKa - Kamchatka R., early run; KB - Bol’shaya R.; KO -OzernayaR.; KP - Palana R. Sampling locations, sampling dates, and number of fish studied Number of fish studied Age DNA Population Group Date of sampling Bio- Electro- determi- extrac- analysis phoresis nation tion Chukotka peninsula, Vaamochka Lake Ch 28 July 2004 50 50 50 50 East Kamchatka, Pakhacha River KPh 17-27 June 2005 67 67 60 55 East Kamchatka, Kamchatka River, late KK 29 June - 9 July 2004 200 200 100 55 run East Kamchatka, Kamchatka River, KKa 3-13 July 2004 150 - 80 60 Azabache Lake, early run West Kamchatka, Ozernaya River KO 4-7 August 2003 200 200 100 100 KB-03 23-30 July 2003 200 200 100 100 West Kamchatka, Bolshaya River KB-04 11-20 August 2004 100 - 80 80 West Kamchatka, Palana River KP 10-21 July 2003 200 200 100 100 West coast of the Sea of Okhotsk, Okh 22 July 2004 100 100 80 70 Okhota River North Kuril Islands, Shumshu Island, 30 July - 5 August Kur 60 60 60 50 Bettobu Lake 2004 DNA extraction, DNA amplification, gel electrophoresis, and band analysis 3’ F 5’ 5’ 3’ DNA R nх230 copies of one DNA 30 cycles F == fragment 3’ 5’ 5’ R 3’ Age structure in sockeye salmon populations studied Freshwater period duration 1 year 1 – 2 years 2 years Bol’shaya R. Chukotka peninsula Ozernaya R. Okhota R. Pakhacha R. Palana R. North Kuril Islands Kamchatka R. Mean size and weight of an individual in sockeye salmon populations 3500 3227 2971 3023 2765 2848 3000 2745 2750 2714 Ch – Chukotka peninsula Kur – North Kuril Islands Weight, g Okh – Okhota R. 2500 2181 Kamchatka peninsula: KPh - Pakhacha R.; 2000 KK - Kamchatka R., late Ch KPh KK KKa KO KB KP Okh Kur run; 66 63.61 KKa - Kamchatka R., 64 61.87 62.14 early run; 60.4 60.84 60.52 60.2 62 KB - Bol’shaya R.; 59.01 58.06 KO - Ozernaya R.; 60 KP -PalanaR. 58 Body length, cm 56 54 Ch KPh KK KKa KO KB KP Okh Kur Multidimensional scaling plot based on biological data Ch KPh Coordinate 2 KK KP KKa KB Okh KO Kur Coordinate 1 Ch – Chukotka peninsula Kur – North Kuril Islands Okh – Okhota R. Kamchatka peninsula: KPh - Pakhacha R.; KK - Kamchatka R., late run; KKa - Kamchatka R., early run; KB - Bol’shaya R.; KO - Ozernaya R.; KP - Palana R. Correlation between genetic and geographic distances among sockeye salmon populations Hypothetic colonization way Fst =0.003ln(D)+0.002 0.06 r=0.24 0.05 Mantel p=0.180 0.04 t s 0.03 F 0.02 0.01 0 369 ln(D) Correlation between genetic and geographic distances among Pacific coast populations (1) and Sea of Okhotsk coast populations (2) Hypothetic colonization way (1) 0.06 0.04 -5 Fst =3·10 D-0.002 st F r=0.71 0.02 Mantel p=0.036 0 0 200 400 600 800 1000 1200 1400 1600 D (2) 0.03 0.02 t F =0.004ln(D)-0.01 s st F r=0.65 0.01 Mantel p=0.504 0 500 1000 1500 2000 D Multidimensional scaling plot based on genetic distances Ch KPh KK Coordinate 2 KP KKa KB Okh KO Kur Coordinate 1 Ch – Chukotka peninsula Kur – North Kuril Islands Okh – Okhota R. Kamchatka peninsula: KPh - Pakhacha R.; KK - Kamchatka R., late run; KKa - Kamchatka R., early run; KB - Bol’shaya R.; KO - Ozernaya R.; KP - Palana R. UPGMA-dendrogram of δμ2 (1) and chord distances (2) for 10 samples of Asian sockeye salmon (1) (2) Ch – Chukotka peninsula Kur – North Kuril Islands Okh – Okhota R. Kamchatka peninsula: KPh - Pakhacha R.; KK - Kamchatka R., late run; KKa - Kamchatka R., early run; KB - Bol’shaya R.; KO - Ozernaya R.; KP -PalanaR. Mean estimated percentage stock compositions of single- population mixtures (correct = 100%) for 9 populations of Asian sockeye salmon Population Estimate Chukotka (Ch) 79.2% Kamchatka, Pakhacha R. (KPh) 80.8% Kamchatka, Kamchatka R., late-run (KK) 74.5% Kamchatka, Kamchatka R., Asabache L., 72.4% early-run (KKa) Kamchatka, Ozernaya R. (KO) 60.0% Kamchatka, Bolshaya R. (KB) 60.8% Kamchatka, Palana R. (KP) 73.7% Okhota R. (Okh) 61.7% North Kuril Islands (Kur) 93.6% 3-D multidimensional scaling plot based on biological and genetic data Ch – Chukotka peninsula 1.5 Kur Ch Kur – North Kuril Islands 1.0 KB Okh – Okhota R. KKa Kamchatka peninsula: .5 KPh - Pakhacha R.; KK - Kamchatka R., late 0.0 run; Coordinate 2 KP -.5 KK KKa - Kamchatka R., KO early run; -1.0 KPh KB - Bol’shaya R.; Okh KO - Ozernaya R.; 2.0 1.5 1.5 2.0 KP -PalanaR. 1.0 .5 .5 1.0 0.0 -.5 -.5 0.0 Coordinate 1 -1.0 -1.0 Coordinate 3 Thank you for your attention!.