Table 1. Geographical locations of Pinus sylvestris populations studied using nuclear and chloroplast microsatellites markers. Native populations are from Poland (PL) divided for three climatic regions: I– Central Poland, II– North Poland, III– South Poland while reference populations are from Finland (F), Turkey (T), Spain (Sp), Scotland (S). Populations from Turkey, Spain and Scotland represent marginal populations, while other samples are from the central pine distribution area.
Code Provenance name Latitude, Longitude Altitude Region
PL1 Karsko 52°54´N, 15°15´E 75 I
PL2 Tabórz 53°52´N, 19°55´E 110 II
PL3 Dłużek 53°31´N, 20°38´E 145 II
PL4 Ruciane 53°37´N, 21°35´E 145 II
PL5 Rozpuda 53°55´N, 22°55´E 205 II
PL6 Supraśl 53°15´N, 23°20´E 165 II
PL7 Starzyna 52°38´N, 23°37´E 170 II
PL8 Gubin 51°55´N, 14°50´E 70 I
PL9 Rychtal 51°12´N, 17°50´E 190 I
PL10 Bolewice 52°23´N, 16°10´E 90 I
PL11 Lipowa 53°44´N, 18°15´E 130 II
PL12 Jegiel 52°40´N, 21°40´E 95 II
PL13 Spała 51°35´N, 20°15´E 150 I
PL14 Janów Lubelski 50°40´N, 22°25´E 250 I
PL15 Nowy Targ 49°20´N, 20°20´E 590 III
PL16 Piwniczna 49°20´N, 20°17´E 500 III
F17 Finland_Joutsa 61°74´N, 26°14´E 125 IV
F18 Finland_Kielajoki 69°65´N, 29°07´E 100
T19 Turkey_Çatacık 39°96´N, 31°11´E 1619 V
T20 Turkey_Bayabat-Sinop 41°64´N, 34°83´E 1228
Sp21 Spain_Sierra de Neila 42°05´N, 03°01´W 1400 VI
Sp22 Spain_Puerto de Navafría 40°98´N, 03°81´W 1800
S23 Scotland 57°30´N, 05°38´W 81 VII
1

Fig. 1. Location of 16 Polish provenances of Scots pine derived from three climatic zones and reference populations of the species distribution range in Europe and Asia. Population acronyms are as in Table 1.

Table 2. Genetic parameters for the nuclear microsatellite loci tested in all Polish pine populations. N – number of alleles per locus, R – range of allele size, A – mean number of alleles per locus, Ae – mean effective number of alleles per locus, Ho – mean observed heterozygosity, He – mean expected heterozygosity, F – mean fixation index, null – null allele frequency, ns – non significant, * P ≤ 0.05.
Locus N R A Ae Ho He F null Deviation from HW equlibrium
psyl42 6 165–177 4.19 3.31 0.673 0.698 0.035 0.000 ns
psyl25Δ 2 213–216 1.56 1.07 0.040 0.062 0.343 0.014 *
psyl18ΔΔ 6 287–305 2.50 1.14 0.119 0.124 0.041 0.000 ns
psyl44 5 166–178 3.06 1.27 0.209 0.215 0.030 0.000 ns
psyl57 7 184–202 5.38 2.35 0.561 0.575 0.024 0.006 *
psyl36 5 245–257 3.56 1.37 0.275 0.270 –0.016 0.000 ns
PtTX4011 10 229–279 5.19 2.94 0.459 0.660 0.304 0.079 *
Spac11.4 21 122–170 10.94 7.84 0.803 0.872 0.079 0.006 *
PtTX3025 10 206–298 5.94 2.61 0.548 0.617 0.112 0.018 *
PtTX4001 16 199–243 7.69 3.37 0.536 0.703 0.237 0.054 *
Δ – locus monomorphic for populations: PL2, PL3, PL5, PL11, PL12, PL14, F17, F18, Sp21, S23
ΔΔ – locus monomorphic for PL11
Table 3. Characteristics of the chloroplast microsatellite loci used to study Scots pine from the Polish species distribution. N – number of alleles per locus, R – range of allele size, A – mean number of alleles per locus, Ae – mean effective number of alleles per locus, uh – unbiased allelic diversity.
Locus N R A Ae uh
PCP36567 4 108–111 2.25 1.40 0.291
PCP30277 8 130–137 5.69 3.68 0.754
PCP102652Δ 4 109–115 1.81 1.10 0.081
PCP26106 4 142–145 2.94 1.71 0.422
PCP87314 4 111–114 3.32 1.51 0.336
PCP45071 6 146–152 3.63 1.75 0.427
PCP1289 4 107–110 3.19 1.75 0.430
PCP41131ΔΔ 5 135–139 2.00 1.23 0.182
Δ – locus monomorphic for populations: PL7, PL8, PL9, PL10,
      PL12, PL14, PL16, F18, F19, Sp21, Sp22, S23
ΔΔ – locus monomorphic for populations: PL10, PL12, PL16, F18
Table 4. Genetic diversity of Scots pine populations in Poland based on nuclear microsatellite loci. A – mean number of alleles per locus, Ae – mean effective number of alleles per locus, Ap – number of private alleles, AR – allelic richness, Ho – mean observed heterozygosity, He – mean expected heterozygosity, F – mean fixation index, LGP – latent genetic potential,
* statistically significant differentiation of populations (P ≤ 0.05).
nSSR diversity
Population Sample size A Ae AP AR Ho He F LGP
PL1 Karsko 19 4.80 2.66 0 4.59 0.496 0.506 0.005 21.42
PL8 Gubin 19 5.30 2.84 0 5.16 0.487 0.492 –0.001 24.61
PL9 Rychtal 24 4.50 2.63 0 4.81 0.388 0.491 0.180 18.70
PL10 Bolewice 21 5.00 2.48 0 4.99 0.407 0.436 0.053 25.20
PL13 Spała 20 5.10 2.72 1 4.41 0.457 0.473 0.010 23.82
PL14 Janów Lubelski 23 4.90 2.41 0 4.83 0.376 0.445 0.108 24.91
PL2 Tabórz 23 4.40 2.45 0 4.76 0.390 0.425 0.051 19.53
PL3 Dłużek 21 4.90 2.39 1 4.20 0.456 0.482 0.037 25.12
PL4 Ruciane 22 5.30 2.84 2 4.86 0.452 0.501 0.062 24.59
PL5 Rozpuda 23 5.00 2.74 0 5.13 0.424 0.464 0.077 22.60
PL6 Supraśl 21 5.10 2.47 0 4.82 0.438 0.465 0.010 26.35
PL7 Starzyna 22 5.50 2.70 1 4.85 0.411 0.461 0.075 27.99
PL11 Lipowa 21 5.10 2.27 2 4.74 0.415 0.445 0.037 28.29
PL12 Jegiel 21 5.10 2.55 0 4.91 0.411 0.471 0.070 25.51
PL15 Nowy Targ 23 5.10 2.24 1 4.64 0.361 0.417 0.085 28.58
PL16 Piwniczna 23 4.90 2.35 0 4.69 0.404 0.417 0.024 25.53
Population means (regions)
I 126 4.9 2.62 1.0 4.80 0.435 0.474 0.059 23.11
II 174 5.1 2.55 6.0 4.78 0.425 0.464 0.052 25.00
III 46 5.0 2.29 1.0 4.67 0.383 0.417 0.054 27.06
Population means (central populations)
Finland 49 5.2 2.49 0.5 4.74 0.484 0.448 –0.049 28.34
Poland 346 5.0 2.55 0.5 4.77 0.423 0.462 0.055 24.55
Populations means (marginal populations)
Turkey 50 4.65 2.35 3 4.42 0.461 0.478 0.069 23.40
Spain 60 4.75 2.46 1 4.37 0.445 0.462 0.079 22.88
Scotland 39 5.70 2.66 1 4.82 0.476 0.482 0.000 30.38
Central vs. marginal populations
Central 395 5.01 2.53 1.3 4.76* 0.435 0.462 0.037 25.03
Marginal 149 5.03 2.49 1.7 4.54* 0.461 0.474 0.049 25.55
Table 5. Genetic diversity of Scots pine populations in Poland based on chloroplast microsatellite loci. Ah – mean number of haplotypes, Ae – mean effective number of haplotypes, AP – number of private haplotypes, AR – haplotype richness, Hd – haplotype diversity, D2sh – mean genetic distance of individuals within populations.
cpSSR diversity
Population Sample size Ah Ae AP AR Hd D2sh
PL1 Karsko 23 20 18.24 7 17.51 0.988 2.518
PL8 Gubin 24 14 10.67 5 11.97 0.946 1.719
PL9 Rychtal 23 21 19.59 3 18.34 0.992 2.015
PL10 Bolewice 24 19 16.94 3 16.20 0.982 1.759
PL13 Spała 22 18 16.13 7 16.36 0.983 2.797
PL14 Janów Lubelski 23 19 16.03 8 16.60 0.980 2.536
PL2 Tabórz 23 20 18.24 9 17.51 0.988 2.875
PL3 Dłużek 21 19 17.64 8 18.00 0.990 2.818
PL4 Ruciane 24 20 18.00 8 16.96 0.986 3.706
PL5 Rozpuda 23 18 14.30 6 15.69 0.972 1.989
PL6 Supraśl 21 21 21.00 11 20.00 1.000 4.089
PL7 Starzyna 21 17 12.60 7 16.00 0.967 1.883
PL11 Lipowa 24 19 16.00 6 16.09 0.978 1.969
PL12 Jegiel 24 19 16.00 2 16.09 0.978 1.879
PL15 Nowy Targ 24 22 20.57 6 18.48 0.993 2.014
PL16 Piwniczna 24 19 14.40 5 15.98 0.971 1.757
Population means (regions)
I 139 18.9 16.53 5.5 16.67 0.981 2.468
II 181 19.2 16.96 7.1 17.36 0.984 2.894
III 48 17.5 15.13 5.5 15.15 0.969 1.867
Population means (central populations)
Finland 49 20.0 17.96 7.3 16.36 0.979 2.041
Poland 368 19.1 16.65 6.3 16.74 0.981 2.395
Populations means (marginal populations)
Turkey 50 18.0 14.66 6.5 14.86 0.970 4.794
Spain 60 21.5 16.80 10.5 15.38 0.972 3.091
Scotland 38 27.0 19.00 11.0 15.74 0.973 2.208
Central vs. marginal populations
Central 417 19.5 16.74 5.5 16.73 0.981 2.346
Marginal 148 21.2 16.38 9.0 15.24 0.971 3.596
Table 6. Hierarchical analysis of molecular variance (AMOVA) at nuclear loci (nSSR diversity) and at chloroplast loci (cpSSR diversity) based on the allelic distance matrix, df – degrees of freedom, SS – sum of squares, MS – mean squares, p – probability.
nSSR diversity cpSSR diversity
Variance component df SS MS Variance % Total p % Total p
Polish populations
Among Regs 2 20.185 10.093 0.016 0 0.095 0 0.165
Among Pops 13 107.865 8.297 0.139 3 0.001 0 0.956
Within Pops 330 1750.996 5.306 5.306 97 0.001 100 0.893
Total 345 1879.046  
Population means (central populations)
Among Regs 3 28.730 9.577 0.013 0 0.120 0 0.507
Among Pops 14 115.596 8.257 0.141 3 0.001 0 0.585
Within Pops 377 1953.509 5.182 5.182 97 0.001 100 0.657
Total 394 2097.835        
Central vs. marginal populations
Among Regs 1 122.613 122.613 0.503 8 0.001 1 0.003
Among Pops 21 258.106 12.291 0.299 5 0.001 0 0.256
Within Pops 521 2761.401 5.300 5.300 87 0.001 99 0.007
Total 543 3142.119        

Fig. 2. a) Principal coordinates plot showing the genetic relationship of the Scots pine populations from the wide distribution of this species. The proportion of variance accounted for by the first two axes was 66.49%, b) Assignment of investigated populations to the two genetic clusters (K = 2) found in the STRUCTURE analysis based on nuclear microsatellite markers. Population acronyms are as in Table 1.