Abstract
The morphological, meiotic and chromosomal variability were studied in two cultivars of Calendula officinalis L. and their mutant lines obtained though chemical mutagenesis using diethyl sulphate (DES) (0.04%, 0.08%) and dimethyl sulphate (DMS) (0.025%, 0.05%). The studied cultivars displayed different sensitivity to DMS and DES mutagens. More M1 plants with morphological changes were observed in C. officinalis cv. ‘Zolotoe more’ than in cv. ‘Rajskij sad’. DMS and DES at low concentrations had positive effects on main agro-metrical traits in both cultivars including plant height, inflorescence diameter and number of inflorescences per plant. Dose-dependent increase in number of various meiotic abnormalities was revealed in both mutant lines. Comparative karyotype analysis and FISH-based visualization of 45S and 5S rDNA indicated a high level of karyotype stability in M1 and M2 plants. Seed treatments with DMS and DES at certain concentrations resulted in higher yields of inflorescences in M1 plants compared to the control. In M2 generation, dose-dependent reduction in the yields of inflorescences was observed. Our findings demonstrate that DMS and DES at low concentrations have great potential in calendula mutation breeding.
Highlights
At the same time, polymorphism in active chemical compounds of the raw material was observed in C. officinalis, and unstable active ingredients, e.g., carotinoids, were revealed[9]
‘Rajskij sad’ had larger inflorescences with bright double orange flowers (Fig. 1). In both studied C. officinalis cultivars, seed treatments with dimethyl sulphate (DMS) and diethyl sulphate (DES) induced a number of morphological changes including plant dwarf, presence of several anthodiums in one inflorescence, polymorphism and fasciation of inflorescences, polypetalous flowers with an increased inflorescence diameter, etc. (Fig. 2)
DMS and DES mutagens stimulated the diversity in morphological characters of C. officinalis including plant height, number of inflorescences, inflorescence diameter, etc
Summary
Polymorphism in active chemical compounds of the raw material was observed in C. officinalis, and unstable active ingredients, e.g., carotinoids, were revealed[9]. As an example, seed treatment with low concentrations of sodium azide and diethyl sulphate was found to influence on seed germination percentage, plant height, leaf area, fresh plant weight, flowering date, inflorescence diameter and gas-exchange measurements in calendula plants of M1 and M2 generations These chemical mutagens had significant effects on total soluble proteins, acid phosphatase, and catalase activity fractions of the calendula leaves[21]. Currently available cytogenetic information regarding this species is rather scarce and mostly obtained by simple chromosome staining It was shown, for example, that a karyotype of C. officinalis was composed by metacentric and submetacentric chromosomes which were small in size (1.5–3.5 μm)[23,24,25]. ‘Rajskij sad’) and their mutant lines (M1 and M2 generations) obtained though chemical mutagenesis using diethyl sulphate (DES) and dimethyl sulphate (DMS) mutagens
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