Indirect in vitro Regeneration of the Medicinal Plant, Aspilia africana, and Histological Assessment at Different Developmental Stages

Abstract

The medicinal plant, Aspilia africana, has been traditionally used in several African countries to treat many diseases such as tuberculosis, cough, inflammation, malaria, osteoporosis, and diabetes. In this study, we developed a protocol for in vitro propagation of A. africana using indirect shoot organogenesis from leaf and root explants of in vitro-grown seedlings and assessed the tissues at different developmental stages. The highest callus induction (91.9 ± 2.96%) from leaf explants was in the Murashige and Skoog (MS) medium augmented with 1.0 mg/L 6-Benzylaminopurine (BAP) and 1.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) while from root explants, the highest callus induction (92.6 ± 2.80%) was in the same plant tissue culture medium augmented with 0.5 mg/L BAP and 1.0 mg/L 2,4-D. The best shoot regeneration capacity from leaf-derived calli (i.e., 80.0 ± 6.23% regeneration percentage and 12.0 ± 6.23 shoots per callus) was obtained in medium augmented with 1.0 mg/L BAP and 0.05 mg/L α-Naphthaleneacetic acid (NAA); the best regeneration capacity for root-derived calli (i.e., 86.7 ± 6.24% shoot regeneration percentage and 14.7 ± 1.11 shoots per callus) was obtained in the MS medium augmented with 1.0 mg/L BAP, 0.05 mg/L NAA, and 0.1 mg/L Thidiazuron (TDZ). Regenerated plantlets developed a robust root system in 1/2 MS medium augmented with 0.1 mg/L NAA and had a survival rate of 93.6% at acclimatization. The in vitro regenerated stem tissue was fully differentiated, while the young leaf tissue consisted of largely unorganized and poorly differentiated cells with large intercellular airspaces typical of in vitro leaf tissues. Our study established a protocol for the indirect regeneration of A. africana and offers a basis for its domestication, large-scale multiplication, and germplasm preservation. To the best of our knowledge, this is the first study to develop an indirect regeneration protocol for A. africana and conduct anatomical assessment through the different stages of development from callus to a fully developed plantlet.

Ploidy Status, Nuclear DNA Content and Start Codon Targeted (SCoT) Genetic Homogeneity Assessment in Digitalis purpurea L., Regenerated In Vitro

“…The results are in accordance with Hesami and Dhaneshvar [ 47 ], who found that 1.5 mg/L BAP along with 0.15 mg/L IBA showed 100% shoot regeneration frequency. Contrary to our observation, BAP with NAA was proved to be an efficient combination for indirect shoot regeneration in Digitalis ferruginea [ 37 ], Rauvolfia serpentina [ 48 ] and Aspilia africana [ 49 ].…”

Phenolic profiling and bioactivity assessment of in vitro propagated Psidium cattleianum Sabine: A promising study

“…In this regard, Rai and his research team [ 30 ] showed that using BA in a concentration of 1 mg/L achieved a high number of buds of P. guajava while increasing the concentration to 2 mg/L induced a sharp decrease in bud formation. Other research groups [ 31 , 32 ] have reported the same observation they found that high concentrations of cytokinin have deleterious effects on the multiplication of shoots in Aspilia africana and Ruta chalepensis , respectively. BA as an adenine cytokinin promotes cell division in plant tissues; hence increasing its level will increase the number of cells, subsequently with cell contents, and the percentage of absorbed water in the cell tissues.…”

Assessment of antibacterial activity and cytotoxic effects of in vitro and in vivo plant parts of a medicinal plant Gynura procumbens (Lour.) Merr.

“…A medicinal plant called Dolichandra unguis-cati (L.) Lohmann showed the best morphogenetic response on MS medium with 0.5 mg/L BAP, and 1.0 mg/L TDZ added in terms of explant response, number of shoots, and length of shoots [ 40 ]. Many other medicinal species have found that full strength of MS medium works best for axillary shoot proliferation, such as G. porcumbens [ 41 , 42 ]; Aspilia africana [ 43 ]; Phaleria macrocarpa [ 44 ].…”