Evaluation of rapid micropropagation of healthy plants of Echinacea purpurea (IDA variety) under influence of cytokinins. Preeliminary study for in vitro protocol

Introduction: Echinacea purpurea (L.) Moench is a valuable ornamental and medicinal plant used in humanand animal medicine. The technique of micropropagation using in vitro plant culture is an promisingalternative to obtain large amounts of healthy plant propagation material in a short time during climatechanges and environmental contamination.Methods: Clonal propagation may be applied for the multiplication of selected genotypes under controlledconditions. This technique also allows acclimating the new plants to elevated temperatures.Objective: The aim of this study was to determine the influence of micodermine, nystatin, miconazoleon purity seed and their germination (n=800, four groups, three replicates). Next aim was to evaluate theinfluence of various concentrations of four cytokinins on induction of adventitious shoots using shoot tips.Results: Benzyloaminopurine (BAP), meta-topolin (mT), and additionally kinetin (KIN), and zeatin (Ze)(1.00-2.00 mg l-1 ) with indole-3-acetic acid (IAA) (0.1-0.2 mg l-1) in Murashige and Skoog (MS) mediumhave been used. Testing the effectiveness of BAP (1.0 mg l-1) and mT (1.0 mg l-1) with IAA (0.1 mg l-1) inMS medium has been caried out for 15 explants in three replicates. For rooting of plants (n=60), IAA atconcentrations of 0.1 mg l-1and 0.5 mg l-1 (three groups in duplicate) has been used. Finally, the effectivenessof acclimatization of rooted plants (n=60) to ex vitro conditions was assessed. All antifungal substanceshave been effective in the obtanment of 100% purity of seeds (n=300). Application of mycodermine resultedin 51% germinating of seeds with signficat differences vs. control seeds (p<0.001), vs. nystatin (p<0.05)and vs. miconazol (p<0.05), and during 16.904.26 days without statisctical differences vs. nystatin and vs.mycodermin. MS basal medium supplemented with BAP (1.0 mg l-1) with IAA (0.1 mg l-1) produced thegreatest number of adventious shoots (n=144), resulting in 9.62.77 shoots per explant on average withsignficat differences vs. control medium MS (p<0.001). However, this result was not statisticaly significantcompared with medium MS supplemented by mT (1.0 mg l-1) with IAA (0.1 mg l-1) (8.933.99 shoots perexplant) and with MS supplemented with BAP (2.0 mg l-1) + IAA (0.2 mg l-1) (7 3.39 shoots per explant).IAA at concentration of 0.1 mg l-1 and 0.5 mg l-1 produced of 100% of rooted plants (n=40) similarly as MSmedium without IAA (n=20) (differences between groups are not statistically significant). Moreover, 70%obtained plants (n=42) have been acclimated to ex vitro conditions using elevated temperature of 35C.Conclusions: Compared to previous study, our experiment showed that low concetration of BAP and mTto effective micropropagation without callus induction can be successfully used. Our results provide theproposal for rapid protocol of clonal propagation of E. purpurea (IDA variety).