In Vitro Adventitious Shoot Regeneration through Direct and Indirect Organogenesis from Seedling-derived Hypocotyl Segments of Ficus religiosa L.: An Important Medicinal Plant

Abstract

<jats:p><jats:italic>Ficus religiosa</jats:italic> is an important industrial, medicinal, and ornamental plant, so in vitro regeneration is of high paramount in this valuable germplasm. Two efficient protocols were developed for indirect and direct shoot organogenesis through hypocotyl explants. In the first experiment, different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) and indole butyric acid (IBA) (0.5, 1.0, and 1.5 mg·L<jats:sup>−1</jats:sup>) in combination with 6-benzyl amino purine (BAP) (ratio 10:1, respectively) were used for callus formation. Two types of callus were obtained from different concentrations of plant growth regulators (PGRs). Also, 2,4-D produced yellow-brownish and friable callus (Type I), whereas the green and compact callus (Type II) was achieved in IBA. The highest callus fresh weight (2.43 g) was observed in Murashige and Skoog (MS) medium containing 0.5 mg·L<jats:sup>−1</jats:sup> 2,4-D plus 0.05 mg·L<jats:sup>−1</jats:sup> BAP. In the later experiments, various concentrations of thidiazuron (TDZ), 6-furfuryl amino purine (KN), and BAP (0.25, 0.5, 1.0, and 1.5 mg·L<jats:sup>−1</jats:sup>) in combination with IBA (ratio 10:1, respectively) were applied for shoot regeneration (direct and indirect organogenesis). In shoot regeneration from callus, the highest regeneration frequency (86.66%) and shoot number per callus (4.13) were achieved in MS medium supplemented with 1.5 mg·L<jats:sup>−1</jats:sup> BAP plus 0.15 mg·L<jats:sup>−1</jats:sup> IBA from type I calli. However, no regeneration was observed in type II calli. In direct shoot regeneration, the highest regeneration frequency (96.66%) and shoot number (6.26) were obtained in the medium mentioned previously. In root induction experiment, different concentrations of naphthalene acetic acid (NAA) and IBA alone or in combination were applied, and MS medium containing 2.0 mg·L<jats:sup>−1</jats:sup> IBA along with 0.1 mg·L<jats:sup>−1</jats:sup> NAA was the best hormonal balance for root induction. The rooted plantlets’ survival rate was more than 95% in the acclimatization stage. These results demonstrated that the direct regeneration method provides more shoot regeneration frequency and take a less time for shoot organogenesis than the indirect regeneration method. Based on our knowledge, this study is the first report of direct and indirect shoot organogenesis of <jats:italic>F. religiosa</jats:italic> via hypocotyl from in vitro–grown seedling.</jats:p>

Modeling and Optimizing in vitro Sterilization of Chrysanthemum via Multilayer Perceptron-Non-dominated Sorting Genetic Algorithm-II (MLP-NSGAII)

“…In general, breeders have made extensive use of the conventional breeding approaches to evaluate desirable traits but still need some techniques due to lack of cross-incompatibility and gene pool resources (Noda et al, 2017). Meanwhile, application of biotechnology in the breeding area could pave the way of evaluating desirable traits by making in vitro regeneration procedures more efficient such as the production of numerous high quality plants in a relatively short time (Naing et al, 2013; Hesami and Daneshvar, 2016, 2018; Hesami et al, 2018a,b). However, contamination during in vitro regeneration procedures is one of the greatest problems that can act as a barrier for making this technique more efficient (Da Silva and Kulus, 2014; Hesami et al, 2017a).…”

Microplants production of Eucalyptus cloeziana from indirect organogenesis

“…This characteristic is important for the plant regeneration, such as reported by Mycock and Watt (2012), when evaluating the callus anatomy in E. grandis W. Hill ex Maiden × E. urophylla S. T. Blake observed that the roots developed from the stem region immediately above the callus or from the callus itself and that of the primordial meristems of the root appear to have been derived from the pericycle of the stem, i.e., from the parenchyma layer between the endodermis and the phloem. The efficiency of in vitro organogenesis depends primarily on the source of the explant, on the components of the culture medium and on the environmental conditions (Hesami andDaneshvar 2018, Silva et al 2019). Another factor influencing the organogenic responses in Eucalyptus is the endogenous and exogenous hormonal balance between the cytokinin and auxin present in the plant tissue, which serve as inducing agents for in vitro morphogenesis (Silva et al 2019, Souza et al 2019.…”

“…Another factor influencing the organogenic responses in Eucalyptus is the endogenous and exogenous hormonal balance between the cytokinin and auxin present in the plant tissue, which serve as inducing agents for in vitro morphogenesis (Silva et al 2019, Souza et al 2019. Many studies report a favorable effect of TDZ at different stages of callus formation (Jafari et al 2017), with TDZ concentrations below 2.5 mg L -1 , being recommended for effective applications to the tissue of woody species, since high concentrations of this plant growth regulator may limit the indirect organogenesis by reducing the induction of buds and increasing hyperhydricity (Hesami and Daneshvar 2018). At 90 days after the start of the experiment, the combination of plant growth regulators and type of explants had an effect on the indirect organogenesis in E. cloeziana.…”

“…The use of indirect organogenesis allows the formation of adventitious bud and shoot elongation for producing large quantities of microplants in a short period and enables the study of different morphogenetic and physiological phenomena (Hesami and Daneshvar 2018), production of secondary metabolites (Mutawil et al 2016), transgenic plants, polyploidy, and somaclonal variations (Hesami and Daneshvar 2018) and callus culture for growing cells in suspension (Silva et al 2019).…”

Morphological Characterization of Ficus religiosa Genotypes in Iran by Multivariate Analysis

“…Ficus is the genus of 1000 species in the family Moraceae, mainly distributed throughout tropical and subtropical regions (Hesami and Daneshvar, 2018). Many of this species have ornamental value and are also used as medicinal plants.…”