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.
Highlights
The plant species Aspilia africana (Pers.) C
The highest percentage of callus induction from the root explants was 92.6 ± 2.80% in the Murashige and Skoog (MS) medium augmented with 0.5 mg/L BAP and 1.0 mg/L 2,4-D, followed by 90.4 ± 3.04%; 87.4 ± 3.57%; 75.6 ± 4.09%, which were all significantly higher (p ≤ 0.05) than the percentage of callus induced in the rest of the treatments (Table 1)
We reported that full-strength MS media augmented with either 1.0 mg/L BAP and 1.0 mg/L Naphthaleneacetic acid (NAA) or 1.0 mg/L BAP and 1.0 mg/L 2,4-D were the most effective for the induction of callus from leaf explants, with a callus induction percentage of over 90%
Summary
The plant species Aspilia africana (Pers.) C. D. Adams, known as the hemorrhage plant or wild sunflower, has been used for several centuries to treat a wide range of health conditions across Africa (Ajeigbe et al, 2014; Okello et al, 2020). The efficacy of A. africana in treating many diseases and managing a range of health conditions is attributed to the abundance of secondary metabolites such as flavonoids, alkaloids, saponins, phenolic compounds, and tannins in the plant (Okoli et al, 2007; Ajeigbe et al, 2014; Komakech et al, 2019)
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