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Healing of cultural varieties paniculate phlox (Phlox paniculata) from nematodes using in-vitro cultures
Nogly, David ; Matiska, Pavel (advisor) ; Augustinová, Ludmila (referee)
Phlox paniculata belongs to the family Poleminiaceae. Phlox paniculata is very important perenial which is used in commons gardens, park landscaping and also to product cut flowers. Although garden phlox are classified as resistant plants to pests and pathogens, they suffer from leaf and stem nematodes of genus Ditylenchus spp. Method eliminating these nematodes from plants for 100 % is still unknown. This thesis investigate the possibility of eliminating nematodes by using explant cultures and creating healthy plants for future mother plants of Phlox paniculata. For this experiment was chosen 10 varieties, 'Aňa Gaganova', 'Blue Paradise', 'Errötense Mädchen', 'Molodost', Igor Talkov', 'Jubilee', 'Laura', 'Mies Copijn', 'Sekret' and 'Winsdor'. These plants were also researched for regeneration of calluses and growing ratio of shoots according to their varietes.
Plants were tested for presence of nematodes Ditylenchus dipsaci by using modified Baermann Technique. All varietes of Phlox paniculata were grown in greenhouse and all of them were tested. For experiment were chosen leaf segments measuring 0,5 - 1 cm2 and the cultivation took place on MS medium (Murashige and Skoog 1962) + 30 g sucrose, pyridoxine 0,5 mg. l-1, thiamine 0,5 mg. l-1, glycine 2 mg. l-1 , nicotinic acid 0,05 mg. l-1. Leaf segments were placed to MS medium enriched by 1,5 mg . l-1 TDZ (Thidiazuron) and 0,5 mg . l-1 IAA (Indole-3-acetic acid). Callus regeneration at the level of plant growth regulators was almost fully vertified during this experiment. Best results were obtained from follow varieties 'Aňa Gaganova', 'Blue Paradise', 'Errötense Mädchen' and 'Molodost'. These results were also obtained from repeated experiment. Positive influence of controlled conditions to growing shoots and callus regeneration was also proved. Explants which were grown in controlled conditions reached higher efficiency on average 0,33 shoots per callus.
By successful recovery of the plants from pests Ditylenchus dipsaci using explantate cultures, it was verifed the initial hypothesis, where resulting explants are transferred to ex-vitro conditions were 100 % pest free. Although, callus cultures grown on MS medium of 1,5 mg . l-1 TDZ and 0,5 mg . l-1 IAA showed some degree of nematode occurrence during testing, consequent passage eliminated the remaining population of nematodes.
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