Abstract
Abstract Leucojum aestivum is a valuable and endangered plant species with bulb scales best suited as explants in micropropagation. In the current study, its micropropagation was investigated by using two different explants and various concentrations and combinations of plant growth regulators (PGRs). Bulbs were first disinfected with benomyl® for 5 hours. After meeting the chilling requirements, two-scale and four-scale explants were provided for direct and indirect organogenesis. Explants were exposed to hot water, 70% ethanol and 2.5% sodium hypochlorite for further disinfestation. Four-scale explants were treated with different concentrations and combinations of naphthaleneacetic acid (NAA), 6-benzyladenine (BA), and kinetin (Kin) for bulblet regeneration. For callogenesis, 0.5 mg L-1 of BA combined with 1, 2, 3, 4, 5 or 6 mg L-1 of 2,4-dichlorophenoxyacetic acid (2,4-D) were applied. Regarding two-scale explants, different combinations and concentrations of BA, Indole-3-butyric acid (IBA) and NAA were used for bulblet induction, and various combinations of Indoleacetic acid (IAA), NAA, 2, 4-D and BA were used for callus induction. None of the two-scale explants responded to the bulblet regeneration and callus induction media. Unlike, four-scale explants regenerated bulblets and roots in the control medium and MS media enriched with different PGRs. Callus was generated on MS medium supplemented with 2,4-D and BA, and indirect regeneration was observed in some cases. On the control medium, the regenerated roots had a natural form, but in PGRs-rich media, they were deformed. Regarding the regeneration percentage, bulblet number and length and root length, no significant differences were found between the control and the best PGR-treatment in each case. Therefore, it seems logical suggesting not to use PGRs, which will considerably reduce the costs at large-scale production.
Highlights
Nowadays, numerous species are in danger of extinction in their natural habitats
During the past three decades, many natural L. aestivum habitats have fallen under threat due to the increasing demand by the pharmaceutical companies (Ivanov et al, 2019), while Ekıcı (2017) claimed that this plant species has the potential for an alternative tourism type, i.e. flora tourism, due to its visual impacts on the landscape
Based on the statistical results, the effect of different plant growth regulators (PGRs)-treatments on four-scale explants in MS medium was significant at 5% for bulblet regeneration rate and number, and in regard to bulblet length, it was significant at 1% level
Summary
Numerous species are in danger of extinction in their natural habitats. Leucojum aestivum L., popularly known as summer snowflake, is an example of valuable plants which, despite its extremely high industrial potential, has received little attention regarding its protection and propagation (Kohut et al, 2007). Member of the Amaryllidaceae family, L. aestivum is a perennial bulbous plant distributed in various countries, including Czechoslovakia, Ireland, Turkey and Iran This plant species is found from east to the west of Iran and south of the Caspian coastal lowlands and north of Iran (Bakian et al, 2020). Because of its attractive flowers, L. aestivum is planted in landscapes, lakes or pools, rock gardens, borders with shrubs and trees (Hundur et al, 2018) This medical species is valuable both in medicine and pharmacy (Cherneva and Dimitrova, 2016) due to alkaloid production, as well as the other plants belonging to the Amaryllidaceae family (Reis et al, 2019)
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