Abstract
Ruta chalepensis L., an evergreen shrub in the citrus family, is well-known around the world for its essential oils and variety of bioactivities, indicating its potential medicinal applications. In this study, we investigated the effect of different culture conditions, including plant growth regulators, media types, pH of the medium, and carbon sources, on in vitro regeneration from nodal explants of R. chalepensis. Following 8 weeks of culture, the highest percentage of regeneration (96.3%) and maximum number of shoots (40.3 shoot/explant) with a length of 4.8 cm were obtained with Murashige and Skoog (MS) medium at pH 5.8, supplemented with 3.0% sucrose and 5.0 µM 6-Benzyladenine (BA) in combination with 1.0 µM 1-naphthaleneacetic acid (NAA). For rooting, individually harvested shootlets were transferred on ½ MS (half-strength) supplemented with IAA (indole-3-acetic acid), IBA (indole 3-butyric acid), or NAA, and the best response in terms of root induction (91.6%), number of roots (5.3), and root mean length (4.9 cm) was achieved with 0.5 µM IBA after 6 weeks. An average of 95.2 percent of healthy, in vitro regenerated plantlets survived after being transplanted into potting soil, indicating that they were effectively hardened. DNA assays (PCR-based markers) such as random amplification of polymorphic DNA (RAPD) and directed amplification of minisatellite-region (DAMD) were employed to assess in vitro cultivated R. chalepensis plantlets that produced a monomorphic banding pattern confirming the genetic stability. Additionally, no changes in the flow cytometric profile of ploidy between regenerated plantlets and donor plants were detected. Regeneration of this valuable medicinal plant in vitro will open up new avenues in pharmaceutical biotechnology by providing an unconventional steadfast system for mass multiplication and might be effectively used in genetic manipulation for enhanced bioactive constituents.
Highlights
Ruta chalepensis L. is a perennial aromatic herb belongs to Rutaceae family, known by the common name “fringed rue” and the Arabic name “Al-Shathap”
Nodal explants cultured on Murashige and Skoog (MS) basal medium failed to respond even after 4 weeks of culture, but MS media supplemented with varying doses of the cytokinins were able to stimulate multiple shoot induction
Analyzing the data (Table 1), the percentage of shoot induction depends on the concentration of cytokinins, and the best results were obtained in the medium containing 5.0 μM BA
Summary
Ruta chalepensis L. is a perennial aromatic herb belongs to Rutaceae family, known by the common name “fringed rue” and the Arabic name “Al-Shathap”. It is spread in the Mediterranean Sea area [1] and widely distributed in the Kingdom of Saudi Arabia. Chalepensis is used in traditional medicine because of its biological activities, which include antioxidant, anti-bacterial [8], anti-fungal [9], anti-inflammatory [1,10], and anthelmintic [11] properties It has been used extensively in folk medicine as an antipyretic and analgesic, as well as for the treatment of mental disorders, convulsions, rheumatism, dropsy, neuralgia, and bleeding problems [4]. It is used internally as an antispasmodic and antihypertensive [1,12,13]
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