engleski

Serological relationships between some Thalictrum species (Ranunculaceae) in Europe

Background and purposes: The taxonomy of genus Thalictrum (Ranunculaceae), especially of T. minus complex, offers a considerable number of unsolved problems. In spite of the accumulation of a large volume of data the systematic and evolutionary relationships between taxa is not clear. First time immunological studies among representatives of several T. minus populations, T. aquilegifolium and T. lucidum in Europe, that aimed at resolving taxonomic problems at subgeneric level are carried out. Materials and methods Proteins were extracted from seed belonging to eight specimens of Thalictrum minus, T. lucidum and T. aquilegifolium species. The colorimetric assay for total protein amount following detergent solubilization was used. The rats were used for antiserum preparation by immunisation three times at 2-week intervals, receiving 100 mg of whole protein concentration subcutaneously for each immunisation. Enzyme-linked immunoadsorbent assay (ELISA) was used to determine serum antibody titters against selected protein antigens. The optical density (OD) of antigen/antiserum reaction was measured at wavelength 405 nm using an automated plate reader. From OD values was calculated Immunological Similarity Index (ISI). ISI matrix was subjected to cluster analysis and Principal Component Analysis. Results and conclusions Currently recognised and separation of T. lucidum and T. aquilegifolium from the members of T. minus complex are supported by serological crossreactivity. The representatives of T. minus complex seem to be clustered in agreement with geographical origin. Average phenetic distance among specimens within T. minus complex indicate that the inter-specific distance is higher than among some member of T. minus complex and well recognised species, i.e. T. aquilegifolim and T. lucidum. The T. minus complex as a whole is as heterogeneous serologically, as it is polymorphic in other aspects.

taxonomy ; serology ; ELISA ; Thalictrum ; Ranunculaceae

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