Towns and cities of Platanthera chlorantha (PS1 and PS2, PB1–PB4, circles) and you may Cephalanthera rubra (CK1 and you can CK2, CB1–CB7, triangles) populations in northern-east Poland.
Research urban area and you can sampling
I examined half a dozen P. chlorantha and you may nine C. rubra communities from inside the north-eastern Poland (Bialowieza and you may Knyszynska Primeval Tree, Szeszupa river area) into the pure, semi-natural and you can anthropogenic groups away from federal and you may surroundings areas, supplies and you can safe components, such as Natura 2000 internet sites ( Fig. 1). And even though they are situated in protected parts, of a lot exist towards railway embankments, collectively channels and you will pathways from inside the forests or perhaps in clearings.
The brand new sampling techniques depended towards the population proportions. Leaf examples off almost all ramets inside populations of any types was basically removed (but society PS2; Desk 1); no trials was basically taken from broken otherwise really more youthful some one. A hundred and ninety-eight trials out-of P. chlorantha and you may 95 trials of C. rubra was in fact gathered. Leaf structure try maintained freeze until it can be held at the ?80 °C, pending allozyme studies. All-collected products were utilized to possess allozyme data.
N, population size; NS, number of samples analysed; NG/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; FIs, 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 loci; A, mean number of alleles per locus; HO, observed heterozygosity; HE, expected heterozygosity; FAre, 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.
Allozyme polymorphism
Homogenates have been made by grinding new will leave in a boundary having 2-mercaptoethanol (1%, v/v). Electrophoresis was accomplished into 10% starch ties in and you can Titan III cellulose acetate plates (Helena Labs, Beaumont, Colorado, USA) adopting the important electrophoretic actions. Ten loci (Adh, Gdh, Got-step 1, Got-2, Idh-step 1, Idh-dos, Mdh-1, Mdh-dos, Me, Pgi, Pgm, 6Pgd, Skd, Sod, Tpi) inside P. chlorantha and you will sixteen loci in C. rubra (Adh, Got-1, Got-2, Gdh, Idh-step one, Idh-dos, Mdh-1, Mdh-2, Myself, 6Pgd, Pgi, Pgm, Skd, Sod, Tpi-1, Tpi-2) had been examined. Several electrode/solution barrier systems were utilized to resolve enzyme systems: GDH and you will Had (10% lithium-borate horizontal starch gel during the pH 8.2/8.3) and you will MDH, SKD and you may TPI (10% histidine-citrate barrier from the pH seven.0/7.0). Enzyme activity staining used Soltis Soltis ( 1989). One other enzyme assistance (ADH, IDH, Me personally, 6PGD, PGI, PGM, SOD) was in fact screened having fun with Titan III cellulose acetate plates, which have been fixed playing with Tris-glycine boundary from the pH 8.6 and Tris-citrate https://datingranking.net/political-dating/ barrier from the pH eight.six (Richardson, Adams Baverstock, 1986). Brand new chemical staining treatments had been centered on Soltis Soltis ( 1989) and you will Richardson ainsi que al. ( 1986), with adjustment.
Mathematical study
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 (FIs actually). 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 FAre) and population size were tested with Spearman’s pairwise rank correlations (StatSoft, 1995).
