The fresh new dating anywhere between parameters off genetic (P

Towns and cities from Platanthera chlorantha (PS1 and PS2, PB1–PB4, circles) and you can Cephalanthera rubra (CK1 and you will CK2, CB1–CB7, triangles) communities for the north-east Poland.

Investigation area and sampling

We investigated six P. chlorantha and you may nine C. rubra communities in the north-east Poland (Bialowieza and Knyszynska Primeval Forest, Szeszupa river valley) for the pure, semi-pure and you can anthropogenic organizations from federal and landscaping parks, supplies and you can safe areas, instance Natura 2000 websites ( Fig. 1). The actual fact that he or she is situated in safe section, of many are present for the railway embankments, with each other channels and you may routes inside forest or in clearings.

The new testing procedure relied into the population proportions. Leaf examples off most ramets within this populations of any kinds was in fact drawn (except people PS2; Table step 1); zero examples was in fact obtained from busted or most more youthful individuals. 100 and you can ninety-7 examples out of P. chlorantha and 95 examples out of C. rubra was in fact built-up. Leaf structure try kept on frost up to it can be stored within ?80 °C, pending allozyme studies. All-collected samples were used to have allozyme investigation.

N, population size; N_S, number of samples analysed; N_Grams/N_W, number of generative ramets/number of vegetative ramets; P_POL, percentage of polymorphic loci; A, mean number of alleles per locus; H_O, observed heterozygosity; H_E, expected heterozygosity; F_Are, inbreeding coefficient; G, number of genotypes, G/N_S, clonal diversity; G_U, number of unique genotypes; G_U %, percentage of unique genotypes (Fischer’s exact test: P < 0.05, **P < 0.01, ***P < 0.001); #, sum of parameters.

N, population size; N_S, number of samples analysed; N_G/N_W, number of generative ramets/number of vegetative ramets; P_POL, percentage of polymorphic loci; A, mean number of alleles per locus; H_O, observed heterozygosity; H_E, expected heterozygosity; F_Is, inbreeding coefficient; G, number of genotypes, G/N_S, clonal diversity; G_U, number of unique genotypes; G_U %, percentage of unique genotypes (Fischer’s exact test: P < 0.05, **P < 0.01, ***P < 0.001); #, sum of parameters.

Allozyme polymorphism

Homogenates had been prepared by grinding the fresh new leaves in a barrier which have 2-mercaptoethanol (1%, v/v). Electrophoresis try carried out on the 10% starch gels and you can Titan III cellulose acetate plates (Helena Laboratories, Beaumont, Tx, USA) after the simple electrophoretic procedures. Fifteen loci (Adh, Gdh, Got-1, Got-dos, Idh-1, Idh-2, Mdh-step 1, Mdh-2, Me personally, Pgi, Pgm, 6Pgd, Skd, Sod, Tpi) into the P. chlorantha and you may 16 loci inside the C. rubra (Adh, Got-step one, Got-2, Gdh, Idh-step one, Idh-2, Mdh-step one, Mdh-dos, Myself, 6Pgd, Pgi, Pgm, Skd, Sod, Tpi-1, Tpi-2) had been examined. A couple electrode/gel barrier options were utilized to resolve chemical assistance: GDH and Had (10% lithium-borate horizontal starch gel during the pH 8.2/8.3) and you can MDH, SKD and you may TPI (10% histidine-citrate boundary during the pH eight.0/eight.0). Chemical activity staining accompanied Soltis Soltis ( 1989). One other chemical solutions (ADH, IDH, Me, 6PGD, PGI, PGM, SOD) was in fact screened playing with Titan III cellulose acetate dishes, that have been resolved playing with Tris-glycine shield at pH 8.6 and you may Tris-citrate boundary in the pH seven.six (Richardson, Adams Baverstock, 1986). Brand new chemical staining pattern have been based on Soltis Soltis ( 1989) and you can Richardson et al. ( 1986), which have adjustment.

Analytical data

The data matrix of individuals was analysed using the TFPGA package (Miller, 1997), FSTAT 2.9.3 (Goudet, 2001) and GENEPOP 3.2 (Raymond Rousset, 1995) for calculation of standard measures of allozyme diversity: allelic frequencies, percentage of polymorphic loci (P_POL), number of alleles per locus (A), genetic diversity (i.e. observed H_O and expected heterozygosity H_E) and inbreeding coefficient (F_Is). The occurrence of unique alleles was used to describe population distinctiveness (Slatkin, 1985). Deviations from hot Buddhist dating Hardy–Weinberg expectations were tested for the population by the Markov chain method (GENEPOP).

Parameters of within-population genotypic diversity were also estimated. Three different measures of clonal diversity were used: number of observed genotypes (G), number of genotypes unique to a single population (G_U) and the probability that the next ramet sampled would be a different genotype (G/N_S; where N_S is the number of ramets sampled). _POL, A, H_O and F_{Is actually}) and population size were tested with Spearman’s pairwise rank correlations (StatSoft, 1995).