Indirect Regeneration and Somatic Embryogenesis from Leaf and Stem Explants of Crassula ovata (Mill.) Druce – An Ornamental Medicinal Plant

This research aims to investigate callus induction, somatic embryogenesis and indirect plant regeneration of Crassula ovata (Mill.) Druce – the famous ornamental plant. Experiment no.1: Callus induction was obtained from leaf and stem explants on Murashige and Skoog (MS) medium supplemented with various plant growth regulators (PGRs). Effects of different PGRs, plant regeneration and subsequent plantlet conversion were also assessed. Indirect plant regeneration was achieved from the callus of stem explants by the addition of 1.5 mg/L Kinetin (KN) alone. Best shoot induction was achieved (6.5 shoots/per explant) after 60 days. For successful rooting, regenerated plantlets were sub-cultured on the same MS media supplemented with 1.5 mg/L KN alone. The rooted plantlets were acclimatized and the survival rate was 90%. Experiment no.2: Results revealed that 0.5 mg/L 2,4-D alone and in combination with 1.0 mg/L 6-Benzyladenine (BA) gave 89.8% callus from the stem explants as compared to leaf explants. Callus proliferation and somatic embryo formation were also evaluated by ‘Double Staining Method’ and different stages of somatic embryogenesis were revealed by scanning electron microscope. Full Strength MS medium produced the highest number (49.6%) of cotyledonary stage somatic embryos (SEs). Mature cotyledonary stage SEs developed into plantlets after 12 weeks of culture. Wellrooted plantlets were successfully acclimatized at the survival rate of 85%. Indirectly regenerated plants did not show any detectable variation in morphological and growth characteristics when compared with the donor plant.

Somatic Embryogenesis for Agropyron cristatum on Murashige and Skoog Medium

Agropyron cristatum L. Gaertn. is a native grass of semiarid region in Iran which is quit resistant to cool and drought climate and withstand heavy grazing. This species has close phylogenetic relationship with Triticum and Hordeum. In this research, the effect of seven different concentrations of growth regulator 2,4-D on callus production and somatic embryogenesis of A. cristatum was investigated on Murashige and Skoog medium. The results showed that the rate of callus, embryo and neomorph were highest in 1 mg L-1 2,4-D. Callus production was increased in 1 mg L-1 2,4-D but dramatically decreased at 5.5 and 9 mg L-1 2,4-D. The somatic embryos were observed at 1 and 4 mg L-1 2,4-D but matured embryos and plantlet were only occurred at 1 mg L-1 2,4-D. There were significant differences between 1 mg L-1 2,4-D and other treatments for producing globular and torpedo embryos, plantlet, rooted callus and number of roots (p

An Efficient Protocol for Cyclic Somatic Embryogenesis in Neem (Azadirachta indica A Juss.)

Neem is a highly heterozygous and commercially important perennial plant. Conventionally, it is propagated by seeds which loose viability within two weeks. Strictly cross pollinating nature of the plant causes serious barrier to the genetic improvement by conventional methods. Alternative methods of tree improvement such as somatic hybridization, mutagenesis and genetic transformation require an efficient in vitro plant regeneration system. In this regard, somatic embryogenesis particularly secondary somatic embryogenesis may offer an effective system for large scale plant propagation without affecting the clonal fidelity of the regenerants. It can be used for synthetic seed production, which further bolsters conservation of this tree species which is otherwise very difficult The present report describes the culture conditions necessary to induce and maintain repetitive somatic embryogenesis, for the first time, in neem. Out of various treatments tested, the somatic embryos were induced directly from immature zygotic embryos of neem on MS + TDZ (0.1 μM) + ABA (4 μM), in more than 76 % cultures. Direct secondary somatic embryogenesis occurred from primary somatic embryos on MS + IAA (5 μM) + GA3 (5 μM) in 12.5 % cultures. Embryogenic competence of the explant as well as of the primary embryos was maintained for a long period by repeated subcultures at frequent intervals. A maximum of 10 % of these somatic embryos were converted into plantlets.

Direct and Indirect Somatic Embryogenesis from Petiole and Leaf Explants of Purple Fan Flower (Scaevola aemula R. Br. cv. 'Purple Fanfare')

Direct and indirect somatic embryogenesis (SE) from petiole and leaf explants of Scaevola aemula R. Br. cv. 'Purple Fanfare' was achieved. High frequency of somatic embryos was obtained directly from petiole and leaf explants using an inductive plant growth regulator signal thidiazuron (TDZ). Petiole explants were more responsive to SE than leaves. Plants derived from somatic embryos of petiole explants germinated more readily into plants. SE occurred more efficiently in half-strength Murashige and Skoog (MS) medium than in full-strength MS medium. Non-embryogenic callus induced by 2, 4-dichlorophenoxyacetic acid was used to investigate the feasibility of obtaining SE with TDZ as a secondary inductive plant growth regulator (PGR) signal. Non-embryogenic callus of S. aemula was able to convert into an “embryogenic competent mode" with PGR signal. Protocol developed for induction of direct and indirect somatic embryogenesis in S. aemula can improve the large scale propagation system of the plant in future.