A Móra Ferenc Múzeum Évkönyve, 1968. (Szeged, 1968)

Gallé, László: The xerothermic lichen species cladonia magyarica VAIN

sugared - the bacteria and penicillia are developing very fast and in a few days they oppress the gonidia propagated much more slowly. In some cultures, anyway, also the gonidia have produced discernible cultures. The necessary inoculations were performed from these cultures. Finally, after repeated inoculations, we have obtained pure cultures, without any bacteria and penicillia. In the last series of culture, the inoculation took place with a micromanipulator. We have striven for getting a pure culture from a single gonidium. For that purpose JANSE-PÉTERFI's micro-manipulator has prover the best. For its use we have applied the method recommended by JAAG (1929, pp. 19-22). Some nice, well-developed thallus squamae of Cladonia magyarica were pured from the contaminations sticked to them and visible even macroscopically. Then they were held under running water for half an hour, and the foreign materials, supposedly sticked to the thallus squamae, incessantly, were washed away with a strong water-spout. After that, the thallus pieces were kept in distilled water sterilized - about ten minutes long - and washed in distilled water with a strong spout. The thallus squamae, rinsed clean in that way, were blotted, placed between elderpith pieces cut in a sterile way, and dried a little. Then, on a slide sterilized in flame, the upper surface of the thallus pieces was scraped somewhat with an annealed lancet. In that way we have obtained the result that the epiphytal algae that sticked to the surface and could not be removed by rinsing, have got into the scrape, as well. And even the upper crust was removed with this method from the layer of gonidia under it. At last, we have separated - similarly with a lancet - the green layer of gonidia from the lower colourless hypha layer. This operation with the moist thallus pieces can be carried out easily. The layer pieces that contained the separeted gonidia were cut up into very tiny pieces on another slide being, similarly sterile. After dropping sterile water on them, we have covered the thallus pieces with a cover-plate and with a mild pressure on the plate we have separated the gonidia and hyphae. After removing the cover-plate, we have obtained water drops containing hypha pieces and free gonidia. We have introduced a small drop of that upon the lower side of the plate covering the wet chamber of the micro-manipulator. In this way we obtained the water drop containing gonidia like a hanging drop. On the right and left sides of the cover plate, beside the water drop with gonidia, we have placed sterile water drops. The drying up of the wet chamber was prevented by the help of a moist wadding. The wet chamber prepared like this was placed on the microscopic slide, and the manipulation was carried out with microscopic magnifications. The pipettes were made according to PÉTERFI's method. We have sucked up a gonidium with a pipette sterilized in absolute alcohol and flame. For removing bacteria or foreign algacells that possibly sticked to it, we have rinsed the gonidium chosen, as well. Therefore, we blew out the content of pipette upon the clean, empty surface of the cover plate, sucking away the water from the gonidium again and again, taking care that the algalcell itself doesn't get into the pipette. Repeating this operation three-four times, we have achieved the result that the selected gonidium cell was available for us clean, in a state suitable for inoculation. With that procedure, which is much more laborious than the former one was, we could inoculate a single gonidium and investigate really the gonidia of Cladonia magyarica in the cultures obtaine by the help of it. An indisputable 242