To determine the minimal DNA sequence homology required for recombination in Bacillus subtilis, we developed a system capable of distinguishing between homologous and illegitimate recombination events during plasmid integration into the chromosome. In this system the recombination frequencies were measured between ts pE194 derivatives carrying segments of the chromosomal beta-gluconase gene (bglS) of various lengths and the bacterial chromosome, using selection for erythromycin resistance at the non-permissive temperature. Homologous recombination events, resulting in disruption of the bglS gene, were easily detected by a colorimetric assay for beta-gluconase activity. A linear dependence of recombination frequency on homology length was observed over an interval of 77 bp. It was found that approximately 70 bp of homology is required for detectable homologous recombination. Homologous recombination was not detected when only 25 bp of homology between plasmid and chromosome were provided. The data indicate that homology requirements for recombination in B. subtilis differ from those in Escherichia coli.
Alfalfa (Medicago sativa; =M. sativa ssp. sativa) in Lithuania is sown as albuminous forage for cattle due to favourable climatic condition. Over many generations, alfalfa plants have escaped from cultivation fields into natural ecosystems and established wild populations. We collected and analyzed individuals from seventeen wild populations of M. sativa. Using random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) analyses, 117 RAPD and 64 ISSR reproducible and highly polymorphic (90.8% for RAPD and 86.3% for ISSR) loci were established. AMOVA showed a high genetic differentiation of M. sativa populations for both types of DNA markers utilized. According to RAPD markers, the genetic variability among populations was 63.1% and 57.0% when ISSR markers were used. Taken together, these results demonstrate that wild populations of M. sativa possess a high potential of genetic variability, that could potentially result in colonization of natural ecosystems. The UPGMA cluster analysis also showed that the DNA markers discovered in this study can distinguish between M. sativa and M. falcata (=M. sativa ssp. falcata) populations and therefore may be used to study the genetic impact of M. sativa on the native populations of M. falcata.
The present study evaluated genetic diversity of Lithuanian populations of Lythrum salicaria in relation to parameters of riparian environment. Growing along Nemunas, Seaside and Lielupė river basins, 15 populations were examined using amplified fragment length polymorphism markers. Molecular data were related to the river basins, type of land use and cover, natural vice versa regulated fragments of the rivers. Population mean genetic diversity parameters were as follows: percentage of polymorphic loci (57.2), expected heterozygosity (0.183), polymorphismc information content (0.218). Mantel test revealed correlation (R2 = 0.0986, p = 0.01) between genetic and geographic distance of populations. Greater genetic diversity within, rather than among populations (ΦPT = 0.213) was observed. According to the Bayesian clustering, studied populations are admixtures of two gene pools. Analysis of molecular variance revealed significant differentiation between populations belonging to distinct river basins, between populations from natural vs. regulated fragments of the rivers.
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