Abstract
Rauvolfia serpentina (L.) Benth. ex Kurz is an endangered medicinal woody species, widely distributed in Asia and used in several traditional medicine systems. Application of in vitro clonal propagation offers alternative strategies for biomass production useful in the production of pharmaceuticals but, difficulty in explant selection and low response to clonal production are impediments to the success. The aim of the present study is to evaluate the efficiency of in vitro rejuvenation of nodal segment explants derived from basal offshoots and terminal buds collected across growth seasons and the effect of serial subcultures on shoot morphogenesis in R. serpentina. Effect of culture medium strength (quarter, half and full-strength Murashige and Skoog medium- MS) on shoot morphogenesis and proliferation through four (4) subcultures were also evaluated. Of the plant growth regulators (PGRs) tested, N6-Benzylaminopurine (BAP) was more efficient (33.87 ± 0.17) over Kinetin (Kin) with 13.92 ± 0.25 and 18.88 ± 0.19 for Thidiazuron (TDZ), and the addition of Naphthalene acetic acid (NAA—0.5 mg L−1) to the PGRs promoted shoot morphogenesis (48.16 ± 0.15, 16.50 ± 0.13 and 26.31 ± 0.17, respectively). Rhizogenesis experiment was carried out using half basal MS medium added with Indole-3-butyric acid (IBA), NAA and Indole-3-acetic acid (IAA). IBA proved the most efficient over the other auxins tested with the production of maximum 12.14 ± 0.24, 9.71 ± 0.31 and 5.26 ± 0.19 roots per micro shoot respectively. However, lower concentration of the IBA showed the most appropriate results on good root differentiation. As a result, IBA at lower concentrations is the most appropriate for good root differentiation. Differential accumulation of pigment molecules and cellular osmolytes in response to the culture condition were also evaluated in the dark-green (DG) and pale-green (PG) leaf morpho-types observed in the shoot cultures; PG leaves showed lower Chla content over the DG ones while Chlb content showed an inverse relationship but, total Chl content was higher in DG leaves (1.20 ± 0.18 mg g−1 FW) over the PG leaves (0.38 ± 0.14 mg g−1 FW). Carotenoids content of the leaves showed a similar trend to the Chl for their complementary photo-protective role to the photosynthetic machinery of the leaves. Accumulation of the osmo-protectants (glycinebetaine, proline and total soluble sugars) showed an inverse trend, with higher values obtained in PG leaves over the DG ones. Results of the present experiment suggest that explant collection season and PGRs influenced in vitro rejuvenation of nodal segment explants through physiological and biochemical changes essential for shoot morphogenesis.
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Abbreviations
- MS:
-
Murashige and Skoog medium
- PGRs:
-
Plant growth regulators
- BAP:
-
N6-benzylaminopurine
- TDZ:
-
Thidiazuron
- Kin:
-
Kinetin
- NAA:
-
Naphthalene acetic acid
- IAA:
-
Indole 3-acetic acid
- IBA:
-
Indole 3-butyric acid
- DG:
-
Dark-green leaves
- PG:
-
Pale-green leaves
- Chl:
-
Chlorophyll
- TSS:
-
Total soluble sugars
- FW:
-
Fresh weight
- PSM:
-
Plant secondary metabolites
- ANOVA:
-
Analysis of variance
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Isah, T., Zafar, N. In vitro rejuvenation of nodal segment explants for clonal propagation of Rauvolfia serpentina (L.) Benth. ex Kurz. Vegetos (2024). https://doi.org/10.1007/s42535-024-00844-0
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DOI: https://doi.org/10.1007/s42535-024-00844-0