Comparison of three chemical tests to assess seed viability: the seed dispersal system of the Macaronesian endemic plant Rubia fruticosa (Rubiaceae) as an example

Summary. Three chemical viability tests were evaluated in the seed dispersal system of Rubia fruticosa, in which three main groups of dispersers participate: reptiles, birds and mammals. Tetrazolium chloride (TTC) and indigo carmine (IC) indicated a lower viability of seeds from droppings of introduced rabbits (Oryctolagus cuniculus) than of those from control plants and the native dispersers, lizards and gulls. In the rabbit seed treatment, significant differences were observed between results obtained with TTC and IC tests. Interpretation of these data, due to the presence of doubtful embryo staining, was more difficult using the IC test. Furthermore, some seeds that were clearly dead had been underestimated. In contrast with results obtained from the two staining methods, the EC test did not confirm that viability of control seeds and those seeds consumed by native dispersers were clearly higher than in seeds ingested by O. cuniculus. Further, compared to the other two tests, the EC method requires more careful handling of the embryo during the extraction process to avoid errors in viability estimation, since this method measures concentration of electrolytes that are released through cellular membranes. Thus, TTC was the most reliable test to assess seed viability in the seed dispersal system of R. fruticosa, and these results agree with those obtained in previous germination experiments made on the same set of seeds given the same treatments.