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The aim of our research was to evaluate the genetic diversity among 25 commercial varieties registered in Poland and 145 landraces of Phaseolus vulgaris var. nanus Asch. (the dwarf common bean) and Phaseolus coccineus L. (the runner bean) maintained in the National Centre of Plant Genetic Resources in Radzików. An additional goal of this study was to compare the precision and efficiency of two techniques of PCR (RAPD and AFLP), used to estimation the genetic diversity of bean. The breeding varieties of bean were registered in the period between 1950 and 2000. The landraces, collected during expeditions conducted from 1985 to 1988, mainly originated from the eastern and southern part of Poland. In the plant genetic diversity research of RAPD and AFLP markers are commonly used. Complex electrophoresis pictures of DNA fragments were taken, and revealed a considerable polymorphism. The polymorphic fragments were obtained on the basis of 6 differentiating primers using the RAPD method and 15 differentiating primers using the AFLP method. P. vulgaris and P. coccineus accessions formed distinct groups. Each of the RAPD and AFLP analyses allowed for the unique distinguishing of all accessions.
Lipoxygenase was localized in the primary leaves of Phaseolus coccineus (L.), seedlings treated with 25 µM Cd and in control plants using the immunogold method. The enzyme was localized mainly in the peripheral parts of protoplast of control plant cells. It was found in the cell wall, along the ER elements, at plastid lamellae and inside the mitochondria. In Cd-treated seedlings the elements of parenchyma cells showed an atypical inner structure. The immunolabelling of LOX was less intensive in comparison with control. The enzyme was found in the cytoplasm, at the cell wall area, vacuoles and in the plastid stroma as single gold particles. LOX activity optima were determined at pH 7.0 and 8.0 for both linoleic and linolenic acid used as substrates. After 2 days of seedlings exposure to Cd the activity of LOX decreased at pH 7.0 and 8.0 when linoleic acid was used as substrate, and strongly declined at pH 7.0 after 4 days of the metal treatment. When linolenic acid was the substrate LOX activity slightly increased after 2 days of the plants exposure to Cd, but after 4 days it rapidly decreased at pH 7.0. The changes in LOX activity are discussed.
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