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ŠUMARSKI LIST 11-12/2019 str. 35     <-- 35 -->        PDF

Embryo and megagametophyte were homogenized in 0.08-0.1 M Tris-HCI buffer with pH 7.5. Other compounds added were: 5 g saccharose, 150 mg DTT and 3 g PVP in 100 ml of buffer. Extracts from the megagametophyte and the embryo were positioned adjacent to each other in the gels. Horizontal starch gel electrophoresis (10.5% starch concentration plus 2.5-3.5% sucrose) was performed as described by Feret and Bergmann (1976), Conkle et al. (1982) and Liengsiri et al. (1990). The buffer system of Ashton pH 8.6 for GOT and PGM as well as Tris-Citro pH 7.3 were used as electrode and gel buffers for SKDH, MDH and 6-PGDH. In addition, used enzyme systems and 10 gene loci are listed in Table 2.
Genetic variation within populations is measured by counting the number of alleles or genotypes per gene locus (AL, GL), with the gene pool diversity in combination with the hypothetical gametic multilocus diversity (v, Vgam) (Gregorius 1978; Gregorius 1987) and the intra-populational genetic differentiation (dT) (Gregorius 1987). Heterozygosity is described by the observed proportion (Ho) of heterozygotes, fixation coefficients according to Wright (1978). Differences between frequencies of genetic types are statistically tested applying the G-test of homogeneity. In the present study, GSED version 1.1 (Gillet 1998) and BIOSYS-2 (Swofford and Selander 1997) programs were used to calculate the measures.