Adeyemi O. Aremu

Somaclonal variation in plants: causes and detection methods

Plant tissue culture has become one of the fundamental tools of plant science research. It is extensively employed in the production, conservation and improvement of plant resources. The presence of somaclonal variation in populations derived from tissue culture is affecting the use of tissue culture negatively and has remained a major problem. Conversely, it is a source of new desirable clones/variants with better agronomic traits. In this review, we summarize the possible causes, detection methods and desirability of variants. Somaclonal variation is one of the most researched and reviewed topics. Hence, we restricted ourselves to outlining various examples which may be used as important references for researchers who intend to identify and/or characterize somaclonal variants while using tissue culture for research and production. Emphasis is placed on the negative effects of somaclonal variation. However, this review also includes examples of some useful variants generated as a result of somaclonal variation.

In vitro antimicrobial, anthelmintic and cyclooxygenase-inhibitory activities and phytochemical analysis of Leucosidea sericea

ETHNOPHARMACOLOGICAL RELEVANCE Leucosidea sericea is used as a vermifuge and in the treatment of ophthalmia by various tribes in southern African countries. AIM OF THE STUDY The study aimed at screening leaves and stems of Leucosidea sericea for pharmacological activity and validating the plants traditional use. A general phytochemical screening was also carried out. MATERIALS AND METHODS Petroleum ether (PE), dichloromethane (DCM), ethanol (EtOH) and water extracts of the plant parts were investigated for antimicrobial, anthelmintic and cyclooxygenase (COX) inhibitory activities. Gram-positive bacteria (Bacillus subtilis and Staphylococcus aureus), Gram-negative bacteria (Escherichia coli and Klebsiella pneumoniae) and Candida albicans were used for the antimicrobial evaluation. Caenorhabditis elegans was used for the anthelmintic assay using the microdilution technique. Cyclooxygenase-1 and -2 (COX-1 and -2) were used to evaluate the anti-inflammatory potential of the plant extracts. Phytochemical analysis for phenolic compounds, including gallotannins, condensed tannins and flavonoids was done using 50% methanol extracts of the leaves and stems employing spectrophotometric methods. RESULTS The leaf extracts exhibited broad spectrum antibacterial activity ranging from 0.025 to 6.25mg/ml. The most noteworthy minimum inhibitory concentration (MIC) of 0.025 mg/ml was exhibited by PE and DCM leaf extracts against Bacillus subtilis and Staphylococcus aureus, respectively. In the anthelmintic assay, the best minimum lethal concentration (MLC) value of 0.26 mg/ml was observed for the DCM and EtOH leaf extracts. Both leaf and stem organic solvent extracts exhibited high to moderate inhibition against COX-1 and -2 at a screening concentration of 250 microg/ml. At lower concentrations, the extracts displayed a dose-dependent inhibition, with the lowest IC(50) values of 0.06 microg/ml (COX-1) and 12.66 microg/ml (COX-2) exhibited by the PE extract of the leaves. Generally, the leaf extracts exhibited better pharmacological activities and contained higher amounts of phenolic compounds than the stem extracts. Alkaloids and saponins were only detected in the leaf and stem extracts, respectively. CONCLUSION The reported results support the local use of Leucosidea sericea against eye infections and as a vermifuge. The pharmacological activities exhibited by the leaf extracts are probably due to their higher phenolic levels.

Isolation of narciprimine from Cyrtanthus contractus (Amaryllidaceae) and evaluation of its acetylcholinesterase inhibitory activity.

ETHNOPHARMACOLOGICAL RELEVANCE Plants of the family Amaryllidaceae are used widely in traditional medicine in South Africa. Several of these, including representatives of the genus Cyrtanthus find use in the treatment of mental illness and age-related dementia. AIM OF THE STUDY Based on the distribution of central nervous system-activating alkaloidal constituents within the genus Cyrtanthus, Cyrtanthus contractus was here explored for such compounds which could interact with acetylcholinesterase (AChE), of significance in the progression of neurodegeneration associated with Alzheimers disease. MATERIALS AND METHODS The known phenanthridone alkaloid narciprimine was isolated via column chromatography of the ethanolic extract of bulbs of Cyrtanthus contractus. The structure of the compound was determined by high field 2D NMR and mass spectroscopic techniques. The classical method of Ellman et al. was used in the determination of AChE inhibitory activity. RESULTS The isolation of narciprimine from Cyrtanthus contractus is a landmark find since it has previously only been identified in Zephyranthes, Narcissus and Lycoris, genera endemic to the Americas, Europe and Asia, respectively. Narciprimine exhibited micromolar inhibitory activity (IC(50) 78.9) against the enzyme acetylcholinesterase. CONCLUSION This work represents the first isolation of narciprimine from an African Amaryllidaceae species, which may be of chemotaxonomic significance. The AChE inhibitory activity of narciprimine, taken together with activities of other structurally similar inhibitors within the series affords further insight to the structural details of the lycorine alkaloid acetylcholinesterase inhibitory pharmacophore.

Antioxidant activity, acetylcholinesterase inhibition, iridoid content and mutagenic evaluation of Leucosidea sericea.

Leucosidea sericea is an important medicinal plant widely used in traditional medicine in southern Africa. Leaf and stem petroleum ether (PE), dichloromethane (DCM) and 50% aqueous methanol (MeOH) extracts were investigated for antioxidant and acetylcholinesterase inhibitory activities. The safety of the extracts was evaluated using the Ames test. In addition, the iridoid content of L. sericea stems and leaves were quantified. For DPPH radical-scavenging activity, the stem MeOH extract (EC(50) value: 1.6 μg/ml) was more potent than ascorbic acid (EC(50) value: 1.7 μg/ml). In the β-carotene-linoleic acid model system, antioxidant activity of the leaf DCM extract (89.8%) was not significantly different to that of butylated hydroxytoluene (BHT) (98.9%). All extracts showed a dose-dependent acetylcholinesterase inhibition; in terms of the IC(50) value, the leaf DCM extract (0.14 mg/ml) was the most potent sample. Total iridoid content was 35% higher in the stem extract than in the leaf extract. Based on the Ames test, L. sericea extracts were not mutagenic, either with or without S9 metabolic activation. These findings suggest the safety as well as the potential of L. sericea as a possible source of novel/alternative antioxidant and acetylcholinesterase inhibitory compounds.

Antioxidant and acetylcholinesterase-inhibitory properties of long-term stored medicinal plants

BackgroundMedicinal plants are possible sources for future novel antioxidant compounds in food and pharmaceutical formulations. Recent attention on medicinal plants emanates from their long historical utilisation in folk medicine as well as their prophylactic properties. However, there is a dearth of scientific data on the efficacy and stability of the bioactive chemical constituents in medicinal plants after prolonged storage. This is a frequent problem in African Traditional Medicine.MethodsThe phytochemical, antioxidant and acetylcholinesterase-inhibitory properties of 21 medicinal plants were evaluated after long-term storage of 12 or 16 years using standard in vitro methods in comparison to freshly harvested materials.ResultsThe total phenolic content of Artemisia afra, Clausena anisata, Cussonia spicata, Leonotis intermedia and Spirostachys africana were significantly higher in stored compared to fresh materials. The flavonoid content were also significantly higher in stored A. afra, C. anisata, C. spicata, L. intermedia, Olea europea and Tetradenia riparia materials. With the exception of Ekebergia capensis and L. intermedia, there were no significant differences between the antioxidant activities of stored and fresh plant materials as measured in the β- carotene-linoleic acid model system. Similarly, the EC50 values based on the 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assay were generally lower for stored than fresh material. Percentage inhibition of acetylcholinesterase was generally similar for both stored and fresh plant material. Stored plant material of Tetradenia riparia and Trichilia dregeana exhibited significantly higher AChE inhibition than the fresh material.ConclusionsThe current study presents evidence that medicinal plants can retain their biological activity after prolonged storage under dark conditions at room temperature. The high antioxidant activities of stable bioactive compounds in these medicinal plants offer interesting prospects for the identification of novel principles for application in food and pharmaceutical formulations.

Antioxidant and phenolic acid profiles of tissue cultured and acclimatized Merwilla plumbea plantlets in relation to the applied cytokinins

Merwilla plumbea (Lindl.) Speta is an important medicinal plant widely used in traditional medicine. We evaluated the effect of five cytokinins [benzyladenine (BA), 2-isopentenyladenine (2iP), meta-topolin (mT), meta-topolin riboside (mTR), and meta-methoxy-9-tetrahydropyran-2-yl-topolin (MemTTHP)] on the level of phenolic acids and antioxidant activity of M. plumbea during the tissue culture and acclimatization stages. Two cytokinins (mT and mTR) significantly improved the antioxidant activity of tissue culture plantlets while the control plantlets were better after acclimatization. Using UPLC-MS/MS, the levels of hydroxybenzoic and hydroxycinnamic acid derivatives (phenolic acids) varied significantly during tissue culture and acclimatization, depending on the cytokinin and plant part analyzed. Vanillic acid (24.9 μg g⁻¹) detected in underground parts of tissue culture plants supplemented with BA was the most abundant phenolic acid detected. The current findings indicate that the phytochemicals together with the bioactivity during in vitro propagation of M. plumbea is influenced by the cytokinin type used and the stage of plant material collection.

The genus Tulbaghia (Alliaceae)—A review of its ethnobotany, pharmacology, phytochemistry and conservation needs

ETHNOPHARMACOLOGICAL RELEVANCE Tulbaghia is a monocotyledonous genus of herbaceous perennial bulbs predominantly found in Africa. The approximately 63 species have nutritive, ornamental and medicinal value. For centuries, several Tulbaghia species have found diverse uses in traditional medicine. AIM OF THE REVIEW To evaluate and document the uses, especially the medicinal value of the genus Tulbaghia. The existing gaps in knowledge and possible means for greater development are highlighted. METHODS A comprehensive and detailed literature search using electronic search (for example, Google Scholar, Scirus and Web of Science) and ethnobotanical books was conducted. RESULTS Findings based on their traditional uses and scientific evaluation indicates that Tulbaghia violacea remain the most widely used species. Compiled data indicate their efficacy in several in vitro and in vivo pharmacological properties such as antimicrobial, anti-hypertensive, antioxidant and anti-cancer activities. Some phytochemicals which are partly associated with the exhibited bioactivities have been identified and isolated from Tulbaghia violacea and Tulbaghia alliacea. The increasing high usage especially the underground organs, as well as the incessant reliance on the wild population, will eventually affect their abundance and possibly make them susceptible to extinction. CONCLUSIONS The great potential of the genus Tulbaghia, particularly other less popular species, remain to be exploited. Further studies on the structural activity relationship of some of the isolated compound may improve their biological potency. Their characteristic alliaceous aroma may also be of great interest to the food industry seeking novel source of unusual aromas to satisfy increasing consumer demands. In order to derive the much anticipated benefits from the species without compromising their conservation status, the use of different biotechnological approaches highlighted in this review will certainly guarantee their continuous availability.

Medicinal plants: An invaluable, dwindling resource in sub-Saharan Africa.

ETHNOPHARMACOLOGICAL RELEVANCE The use of plant species for different therapeutic/medicinal purposes is well-entrenched in sub-Saharan Africa. AIM OF THE REVIEW To provide a critical and updated review of the enormous medicinal plant heritage in sub-Saharan Africa with regards to the abundance, importance, conservation status and potential means to help sustain their availability for future generations. METHODS A comprehensive literature search involving different online databases, books and theses were conducted in order to obtain, collate and synthesize available information on various fundamental aspects pertaining to African medicinal plants. RESULTS African biodiversity hotspots are endowed with a high level of endemic species with a significant portion possessing medicinal value. Apart from the extensive ethnobotanical uses of medicinal plants found in Africa, scientific validation of their biological potential such as antimicrobial, antioxidant, anti-inflammatory and anti-diabetic properties have been recognized. Together with the demand arising from their biological efficacies, other anthropogenic factors are exerting conservation strains of the wild population of these medicinal plants. Even though researchers have acknowledged the importance and value of conserving these medicinal plants, several challenges have hampered these efforts on the Continent as a whole. CONCLUSIONS The rich flora occurring in sub-Saharan Africa suggests enormous potential for discovery of new chemical entity with therapeutic value. However, concerted efforts focused on documenting the conservation status of African medicinal plants are pertinent. Application of different biotechnological techniques is needed to sustain these valuable botanical entities, especially to meet increasing pharmaceutical demand. Most importantly, increased public enlightenment and awareness may help eradicate the prejudice against cultivation of medicinal plants.

Shoot proliferation and rooting treatments influence secondary metabolite production and antioxidant activity in tissue culture-derived Aloe arborescens grown ex vitro

Aloe species are valuable plants with great ornamental and medicinal value. Although micropropagation protocols have been developed to meet the increasing global demand, the effects of the series of events during micropropagation on the phytochemical and pharmacological efficacy of ex-vitro plants remains poorly understood. Thus, we evaluated the effects of cytokinin and rooting compounds used during the shoot regeneration and rooting phases respectively, on secondary metabolite production in greenhouse-grown in vitro-derived Aloe arborescens. Shoots derived from meta-methoxytopolin (MemT)-containing medium and rooted with either smoke–water (SW) or indole butyric acid (IBA) had higher levels of total phenolics and flavonoids than those rooted on plant growth regulator (PGR)-free medium. Iridoid content was significantly reduced in cytokinin-regenerated shoots rooted with IBA in comparison to PGR-free regenerated shoots rooted with IBA. Conversely, the use of SW for rooting in cytokinin-regenerated shoots significantly increased iridoid content when compared to PGR-free regenerated shoots rooted with SW. These findings suggest an antagonistic interaction between cytokinins used in this study and IBA as well as a possible synergistic or additive interaction of the cytokinins with SW on iridoid production. Significantly higher antioxidant activity was recorded in shoots regenerated from meta-topolin riboside (mTR) and MemT and rooted with IBA or SW when compared to those rooted without PGR. Overall, the type of cytokinin and rooting treatments individually and interactively had a significant carry-over effect on secondary metabolite production and antioxidant potential of tissue culture-derived A. arborescens. Therefore, when micropropagating plants for medicinal uses, it is prudent to select the right cytokinin and rooting compound for optimal production of secondary metabolites and ultimately the pharmacological efficacy of acclimatized plants.

Conservation strategy for Pelargonium sidoides DC: phenolic profile and pharmacological activity of acclimatized plants derived from tissue culture.

ETHNOPHARMACOLOGICAL RELEVANCE Pelargonium sidoides DC (Geraniaceae), a popular medicinal plant used in folk medicine in the treatment of respiratory-related infections has gained international prominence due to its usage in several herbal formulations. This has led to high demand and the subsequent decimation of wild populations. AIM OF THE STUDY Using plant tissue culture techniques, Pelargonium sidoides plants were cloned in vitro, acclimatized under greenhouse conditions and evaluated for their phytochemical content and pharmacological activity. METHODS Phenolic content in extracts of in vitro-derived, greenhouse-acclimatized and wild Pelargonium sidoides plants were analyzed using UPLC-MS/MS. The oxygen radical absorbance capacity (ORAC), 2,2-diphenyl-1-picryl hydrazyl (DPPH) radical scavenging activity and minimum inhibitory concentration (MIC) of the extracts against bacterial and fungal strains were evaluated. RESULTS Similarities in phenolic profiles were identified confirming the chemical signatures that characterize Pelargonium sidoides plants. Extracts of greenhouse-acclimatized and wild plants exhibited comparable antimicrobial and antioxidant properties. CONCLUSIONS Overall, the study highlights the potential of integrating plant tissue culture technologies in conservation strategies of medicinal plants. In particular, the results strongly suggest the feasibility of both large-scale cultivation and plant part substitution as alternative solutions to the current destructive overharvesting practices of wild Pelargonium sidoides populations.