Regeneration of plantlets through somatic embryogenesis from root derived calli of Hibiscus sabdariffa L. (Roselle) and assessment of genetic stability by flow cytometry and ISSR analysis

Abstract

Induction of somatic embryogenesis and complete plantlet regeneration from callus culture of Hibiscus sabdariffa L. var. HS4288 has been made. Leaf and root explants were cultured on Murashige and Skoog (MS) and Driver–Kuniyuki Walnut (DKW) basal media supplemented with different concentrations of synthetic auxins and cytokinins. Root explants on DKW medium supplemented with 2.26μM 2, 4-Dichlorophenoxyacetic acid (2, 4-D) and 4.65μM kinetin (KIN) induced highest percentage (70%) of embryogenic calli. Average number of globular embryos per root derived callus produced within 6 weeks of culture initiation on MS media with different plant growth regulators (PGRs) ranged from 2.27±0.12 to 8.80±0.17 and that of cotyledonary embryos ranged from 0.00 to 2.53±0.20. On DKW medium comparatively more globular embryos (2.70±0.15 to 14.53±0.23) and cotyledonary embryos (0.00 to 8.90±0.17) were produced than that of MS medium. Regeneration of complete plantlets was highest (76.67%) when embryogenic calli with mature somatic embryos were grown on DKW medium containing 2.32μM KIN and 2.22μM 6-Benzyladenine (BA). Plants were primarily hardened in humidity, temperature and light controlled chamber and finally in a greenhouse showed 70% survival ability. Different stages of somatic embryogenesis process in the root derived embryogenic calli were elaborated in detail by morphological, histological and SEM study. The data were statistically analyzed by Duncan Multiple range test (p ≤ 0.05) and Principal component analysis (PCA). Flow cytometry and Inter-simple sequence repeats (ISSR) marker analysis confirmed that there was no genetic variation within the regenerated plants.

Determination of Genetic Stability in Cacao Plants (Theobroma Cacao L.) Derived from Somatic Embryogenesis Using Microsatellite Molecular Markers (SSR)

“…This result supports the conclusion that the regenerated CCN51 and TSH565 plants produced via somatic embryogenesis had no variations with respect to field plants. Similar results, where a genetic homogeneity of close to 100%, have been obtained from other species derived from somatic embryogenesis, both directly and indirectly, from Hibiscus sabdariffa L (Konar et al, 2018), Abutilon indicum L. (Seth et al, 2017), Cucumis melo L. (Raji et al, 2018), Anoectochilus elatus Lindl (Sherif et al, 2018), Citrullus lanatus (Vinoth and Ravindhran, 2016) and Bacopa monnieri (Largia et al, 2015).…”

Development and regeneration of somatic embryos from leaves-derived calli of Coffea liberica

“…This showed that the somatic embryo development process is asynchronous. Similar results have been found in Hibiscus sabdariffa (Konar et al 2018), Akebia trifoliata (Zou et al 2019), Helianthus annuus L. (Fu et al 2017), and Capsicum baccatum (Venkataiah et al 2016). Finally, when the cotyledonary stage embryos were sub-cultured in MS medium containing 1 mgL -1 BAP at 12 WAC, they were converted into complete plantlets (Figure 1.G).…”

Secretome-Derived Cultured Cell System: Overview Towards Extracellular Protein Characterization and Biotechnological Applications

“…SE is already known as the most appropriate regeneration system based on the exceptional totipotency of pro-embryonic masses (PEMs) giving rise to hundred of somatic embryos able to regenerate into plants. Several explants have been used to induce PEMs including immature flower tissues [7], anthers [7,8], ovaries [7,9], microspores [10], mature and immature zygotic embryos [9,11], cotyledons [12,13], hypocotyls [13], petioles [14], root tip [15,16], shoot apex [17] and leaf explants [16,18]. Somatic embryos developed over the surface of embryogenic callus (indirect SE) or occasionally, directly from explants without intervening calls phase (direct SE).…”