This new matchmaking ranging from parameters away from genetic (P
Metropolitan areas regarding Platanthera chlorantha (PS1 and you may PS2, PB1–PB4, circles) and you may Cephalanthera rubra (CK1 and you may CK2, CB1–CB7, triangles) communities during the northern-east Poland.
Studies area and you will sampling
We examined half a dozen P. chlorantha and you will 9 C. rubra populations inside north-east Poland (Bialowieza and you may Knyszynska Primeval Tree, Szeszupa river area) into the sheer, semi-absolute and anthropogenic groups out-of national and you may land parks, reserves and protected section, eg Natura 2000 internet ( Fig. 1). Despite the reality they are located in secure portion, many exists towards railway embankments, collectively roads and you can pathways into the woods or in clearings.
This new testing processes relied into the population proportions. Leaf products of almost all ramets inside populations of any types was in fact removed (except population PS2; Desk step one); zero products have been extracted from damaged or extremely more youthful someone. A hundred and you can 90-seven samples away from P. chlorantha and you may 95 products of C. rubra have been amassed. Leaf cells was maintained freeze up until it may be held on ?80 °C, pending allozyme analysis. All-collected products were used for 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 loci; A, mean number of alleles per locus; HO, observed heterozygosity; HE, expected heterozygosity; FTry, 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 had been made by grinding the fresh new leaves for the a buffer which have 2-mercaptoethanol (1%, v/v). Electrophoresis try accomplished with the ten% starch ties in and you will Titan III cellulose acetate dishes (Helena Laboratories, Beaumont, Colorado, USA) following basic electrophoretic methods. Fifteen loci (Adh, Gdh, Got-step one, Got-dos, Idh-step one, Idh-2, Mdh-step 1, Mdh-dos, Me personally, Pgi, Pgm, 6Pgd, Skd, Sod, Tpi) into the P. chlorantha and sixteen loci from inside the C. rubra (Adh, Got-step one, Got-2, Gdh, Idh-step one, Idh-2, Mdh-step 1, Mdh-dos, Me, 6Pgd, Pgi, Pgm, Skd, Sod, Tpi-step 1, Tpi-2) was investigated. Several electrode/gel barrier assistance were used to respond to chemical expertise: GDH and you may Had (10% lithium-borate lateral starch gel at pH 8.2/8.3) and you may https://datingranking.net/equestrian-dating/ MDH, SKD and you can TPI (10% histidine-citrate shield within pH eight.0/7.0). Enzyme passion staining used Soltis Soltis ( 1989). Additional chemical expertise (ADH, IDH, Me personally, 6PGD, PGI, PGM, SOD) was in fact screened having fun with Titan III cellulose acetate dishes, that have been fixed having fun with Tris-glycine boundary at pH 8.6 and you can Tris-citrate shield on pH 7.six (Richardson, Adams Baverstock, 1986). The new enzyme staining treatments was in fact according to Soltis Soltis ( 1989) and you will Richardson ainsi que al. ( 1986), having modifications.
Mathematical investigation
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).
