14 Jun The fresh new relationships between variables out-of genetic (P
Cities from Platanthera chlorantha (PS1 and you will PS2, PB1–PB4, circles) and you can Cephalanthera rubra (CK1 and you will CK2, CB1–CB7, triangles) communities in the north-east Poland.
Studies area and testing
We investigated half dozen P. chlorantha and you will 9 C. rubra communities within the northern-eastern Poland (Bialowieza and you may Knyszynska Primeval Forest, Szeszupa lake valley) in the sheer, semi-pure and anthropogenic organizations regarding national and you will landscaping areas, supplies and you can secure components, particularly Natura 2000 internet ( Fig. 1). Though he’s located in protected components, of a lot can be found towards rail embankments, collectively routes and paths from inside the forests or perhaps in clearings.
The fresh sampling techniques depended on people dimensions. Leaf trials regarding most ramets within this communities each and every varieties had been removed (but people PS2; Table step 1); no examples was in fact taken from busted otherwise extremely younger individuals. One hundred and you can ninety-seven examples regarding P. chlorantha and you can 95 trials away from C. rubra was indeed collected. Leaf structure try maintained frost up to it may be stored within ?80 °C, pending allozyme research. All collected samples were utilized to own allozyme analysis.
N, population size; NS, number of samples analysed; NGrams/NW, number of generative ramets/number of vegetative ramets; PPOL, percentage of polymorphic loci; A, mean number of alleles per locus; HO, observed heterozygosity; HE, expected heterozygosity; FWas, inbreeding coefficient; G, number of genotypes, G/NS, clonal diversity; GU, number of unique genotypes; GU %, percentage of unique genotypes (Fischer’s exact test: P < 0.05, **P < 0.01, ***P < 0.001); #, sum of parameters.
N, population size; NS, number of samples analysed; NG/NW, number of generative ramets/number of vegetative ramets; PPOL, percentage of polymorphic Sugar Momma Sites dating loci; A, mean number of alleles per locus; HO, observed heterozygosity; HE, expected heterozygosity; FIs actually, inbreeding coefficient; G, number of genotypes, G/NS, clonal diversity; GU, number of unique genotypes; GU %, percentage of unique genotypes (Fischer’s exact test: P < 0.05, **P < 0.01, ***P < 0.001); #, sum of parameters.
Homogenates was served by grinding this new departs within the a boundary that have 2-mercaptoethanol (1%, v/v). Electrophoresis are carried out with the 10% starch gels and you can Titan III cellulose acetate plates (Helena Labs, Beaumont, Texas, USA) following the important electrophoretic strategies. Ten loci (Adh, Gdh, Got-1, Got-2, Idh-1, Idh-2, Mdh-1, Mdh-2, Myself, Pgi, Pgm, 6Pgd, Skd, Sod, Tpi) inside P. chlorantha and sixteen loci in the C. rubra (Adh, Got-1, Got-2, Gdh, Idh-step one, Idh-2, Mdh-1, Mdh-2, Me personally, 6Pgd, Pgi, Pgm, Skd, Sod, Tpi-step one, Tpi-2) had been investigated. One or two electrode/solution shield options were used to answer chemical possibilities: GDH and you may Had (10% lithium-borate lateral starch gel within pH 8.2/8.3) and you may MDH, SKD and TPI (10% histidine-citrate buffer within pH 7.0/eight.0). Chemical passion staining accompanied Soltis Soltis ( 1989). The other enzyme expertise (ADH, IDH, Myself, 6PGD, PGI, PGM, SOD) have been processed having fun with Titan III cellulose acetate dishes, which were resolved having fun with Tris-glycine barrier at the pH 8.6 and you can Tris-citrate boundary at pH 7.6 (Richardson, Adams Baverstock, 1986). This new enzyme staining formulas was basically centered on Soltis Soltis ( 1989) and you can Richardson et al. ( 1986), having modifications.
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 (PPOL), number of alleles per locus (A), genetic diversity (i.e. observed HO and expected heterozygosity HE) and inbreeding coefficient (FAre). The occurrence of unique alleles was used to describe population distinctiveness (Slatkin, 1985). Deviations from 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 (GU) and the probability that the next ramet sampled would be a different genotype (G/NS; where NS is the number of ramets sampled). POL, A, HO and FIs actually) and population size were tested with Spearman’s pairwise rank correlations (StatSoft, 1995).