Comparison of the Effect of Meta-Topolin and Benzyladenine during Daphne mezereum L. Micropropagation

Karolina Nowakowska,Andrzej Pacholczak

doi:10.3390/agronomy10121994

Karolina Nowakowska, Andrzej Pacholczak

Open Access

https://doi.org/10.3390/agronomy10121994

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Abstract

Meta-Topolin (mT) is one of the lesser-known and only recently discovered cytokinins that can be used as an alternative to the commonly used benzyladenine (BA). The experiment aimed to compare the effects of BA and mT on the regeneration from explants of Daphne mezereum, on the levels of several chemical components, the catalase activity, and to assess genetic diversity of microcuttings. The Murashige and Skoog medium (MS) and the woody plant medium (WPM) were used, enhanced with auxin and 1 mg·L−1 mT or 1 mg·L−1 BA. The results suggest that mT has a more positive effect on shoot regeneration. It also positively affected the levels of various compounds which are generally taken as indicative of micro cutting conditions. The levels of chlorophyll and carbohydrates were the highest in microcuttings from the WPM with 0.1 mg·L−1 NAA. Total soluble proteins and amino acids were the highest in plants from the MS medium with 1 mg·L−1 mT. Increased levels of hydrogen peroxide and catalase activity were observed on media with 1 mg·L−1 BA. Screening with 30 randomly amplified polymorphic DNAs (RAPD) and 20 inter-simple sequence repeat (ISSR) primers demonstrated a considerable level of genetic variation among regenerating plants. This variation is attributed to the condition of the starting material (seed propagated); there were no indications that the two cytokinins tested contributed to increased somaclonal variation.

Highlights

February daphne, Daphne mezereum L., belongs to the Thymelaeaceae family of plants
20 inter-simple sequence repeat (ISSR) primers demonstrated a considerable level of genetic variation among regenerating plants
This variation is attributed to the condition of the starting material; there were no indications that the two cytokinins tested contributed to increased somaclonal variation

Summary

Introduction

Daphne mezereum L., belongs to the Thymelaeaceae family of plants. In Poland, the shrub is legally protected. It needs protection as its habitat, deciduous forests, is being replaced by conifers. Its natural stands are often vandalized by breaking off the flowering branches or digging bushes out for transfer into gardens [1]. Apart from flowers, which appear very early in spring, ornamental features of the plant include ornamental red berries of 1 cm in diameter which ripen at the end of June. The shrub is mainly used in gardens, on flower beds, and in rock gardens [2]

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