In vitro propagation of Disanthus cercidifolius and assessment of genetic fidelity of regenerants

Bc. Tereza Ulvrová

Master's thesis

Abstract:

In this study, an efficient micropropagation protocol has been developed for Disanthus cercidifolius Maxim. (Hamamelidaceae), a shrub originating in China and Japan, with attractive purple fall colour of leaves and great horticultural potential as ornamental shrub in parks and gardens. For this study, sprouting buds of two mature shrubs (genotype Truba and genotype PdS) were used as an initial plant material. For the in vitro propagation experiment, the nodal segments were cultured on a MS medium (Murashige and Skoog 1962) supplemented with either BAP (0.5 - 3 mg/l) or zeatin (0.5 - 3 mg/l). As control, MS medium without plant growth regulators was used. The highest number of shoots per explant (in average 6.95 in genotype PdS and 7.93 in genotype Truba, respectively) was achieved on a medium supplemented with 2 mg/l BAP. Two types of media, half-strength MS medium and half-strength WPM medium (Lloyd and McCown, 1980) supplemented with IBA (0.1 - 0.5 mg/l), were tested for rooting of the shoots resulting in the best rooting performance on a half-strength WPM medium containing 0.5 mg/l IBA. The rooted plantlets were transferred ex vitro, with 60% survival rate in genotype PdS and 33,3% in genotype Truba. Inter simple sequence repeat (ISSR) and flow cytometry were used in three shoots per 6 randomly chosen regenerants and two control plants (one for each genotype) to assess their genetic fidelity. Ten ISSR primers were used for molecular analysis and all amplified products were monomorfic indicating no genetic variability. Similarly, flow cytometric analysis showed that the ploidy level in all in vitro regenerants was stable and identical to mother plants. This analysis confirmed that micropropagation protocol optimized here represents a reliable method for the production of genetically uniform plants. Therefore, after further improvement of ex vitro transfer, the protocol described here could be utilized for effective mass propagation of Disanthus cercidifolius.

Keywords

Disanthus cercidifolius, genetic fidelity, in vitro propagation, molecular markers, plant growth regulators

Language used: English

Date on which the thesis was submitted / produced: 27. 4. 2017

Thesis defence

• Supervisor: Ing. Iva Viehmannová, Ph.D.

Czech University of Life Sciences Prague

Faculty of Tropical AgriSciences

Master programme / field:
Tropical Agriculture / Tropical Crop Management and Ecology