Micropropagation and secondary metabolites of Sclerocarya birrea.

Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2009.Sclerocarya birrea (marula, Anacardiaceae) is a highly-valued indigenous tree in most parts of sub-Saharan Africa because of its medicinal and nutritional properties. The marula tree is adapted to the semi-arid conditions that characterise most parts of sub-Saharan Africa and renders them unsuitable for conventional crop agriculture. The unique nutritional properties of marula and its high tolerance to dry conditions provide opportunities for its development into a plantation crop. On the other hand, the demand for marula plant parts, mainly the bark and roots as medicinal remedies, poses a great threat to wild populations. In the long term, the growing demand of marula products in the food, pharmaceutical and cosmetic industries will not be sustainable from wild populations alone. Plant tissue culture technologies can be useful for in vitro manipulation and mass propagation of the plant in the process of domestication and conservation. The aims of the project were to determine the optimum conditions for seed germination, in vitro propagation and plant regeneration, and to evaluate the potential bioactivity of secondary metabolites from its renewable plant parts as an alternative option in the conservation of S. birrea. An ex vitro seed germination study indicated that after-ripening and cold stratification are critical factors. Cold stratification (5 °C) of marula nuts for 14 days improved germination (65%) as compared to non-stratified nuts (32%). Direct shoot organogenesis was achieved from leaf explants through the induction of nodular meristemoids on Murashige and Skoog (MS) (1962) medium and woody plant medium (WPM) supplemented with 6-benzyladenine (BA) in combination with naphthalene acetic acid (NAA), indole-3-butryric acid (IBA) and indole-3-acetic acid (IAA). Induction of nodular meristemoids from 86% of the leaf cultures was achieved on a MS medium with 4.0 ìM BA and 1.0 ìM NAA. High levels (78–100%) of induction were also achieved on WPM with different concentrations of BA (1.0–4.0 ìM) and IBA (1.0–4.0 ìM). The highest conversion of nodular meristemoids into shoots on MS initiation medium was only 22% for 4.0 ìM BA and 1.0 ìM NAA. This was improved to 62% when nodular clusters were cultured in MS liquid medium. Histological studies revealed high numbers of unipolar meristematic buds developing from globular nodules. These embryo-like structures have in the past been mistaken for true somatic embryos. The initiation of high numbers of nodular meristemoids per explant provides potential for automated large-scale clonal propagation in bioreactors, in vitro phytochemical production and the development of synthetic seed technology, similar to somatic embryogenesis. Plant regeneration through nodule culture has potential for application in mass micropropagation and plant breeding of S. birrea. Adventitious shoot and root induction are important phases in micropropagation. Plant growth regulators play an important role in these developmental processes, and the type and concentration used have major influences on the eventual organogenic pathway. Three auxins (IAA, IBA and NAA) and four aromatic cytokinins (6-benzyladenine, meta-topolin, meta-topolin riboside, and meta-methoxytopolin riboside) were evaluated for their potential to induce adventitious shoot and root formation in S. birrea shoots, hypocotyls and epicotyls. Among the evaluated cytokinins, the highest adventitious shoot induction (62%) was achieved on MS medium supplemented with meta-topolin (8.0 ìM). The lowest adventitious shoot induction (2.5%) was obtained on MS basal medium containing 2.0 ìM meta-methoxytopolin riboside. The highest adventitious shoot induction for hypocotyls was 55% on MS medium supplemented with 8.0 ìM meta-topolin. For the tested auxins, IBA induced adventitious rooting in 91% of shoots at a concentration of 4.0 ìM after 8 weeks in culture. However, the in vitro rooted plants only survived for two weeks when transferred ex vitro. A temperature of 25 °C and 16-h photoperiod were optimum for adventitious root induction. Stomatal density (number per mm2) on the abaxial leaf surfaces was higher for the 16-h photoperiod treatment (206.6 ± 15.28) compared to that for a 24-h photoperiod (134.6 ± 12.98). Normal mature stomata with kidney-shaped guard cells and an outer ledge over the stomatal pore were observed for in vitro plants growing under a 16-h photoperiod. Total phenolic content, proanthocyanidins, gallotannins, flavonoids, and antioxidant activities of S. birrea methanolic extracts were evaluated using in vitro bioassays. Methanolic extracts of the young stem bark and leaves contained high levels of these phytochemicals. Sclerocarya birrea young stem extracts contained the highest levels of total phenolics (14.15 ± 0.03 mg GAE g-1), flavonoids (1219.39 ± 16.62 ìg CE g-1) and gallotannins (246.12 ± 3.76 ìg GAE g-1). Sclerocarya birrea leaf extracts had the highest concentration of proanthocyanidins (1.25%). The EC50 values of the extracts in the DPPH free radical scavenging assay ranged from 5.028 to 6.921 ìg ml-1, compared to ascorbic acid (6.868 ìg ml-1). A dose-dependent linear curve was obtained for all extracts in the ferric-reducing power assay. All the extracts exhibited high antioxidant activity comparable to butylated hydroxytoluene based on the rate of â-carotene bleaching (89.6 to 93.9%). Sclerocarya birrea provides a source of secondary metabolites which have potent antioxidant properties and may be beneficial to the health of consumers. Sclerocarya birrea young stem and leaf ethanolic extracts exhibited high bioactivity (MIC < 1.0 mg ml-1) against both Gram-positive (Bacillus subtilis and Staphylococcus aureus) and Gram-negative (Escherichia coli and Klebsiella pneumoniae) bacteria. The highest activity (MIC = 0.098 mg ml-1 and total activity = 1609.1 ml g-1) was recorded for young stem extracts against B. subtilis. The highest activity (MIC = 1.56 mg ml-1 and MFC = 1.56 mg ml-1) in the antifungal assay against Candida albicans was observed for young stem ethanolic extracts. Sclerocarya birrea extracts had moderate acetylcholinesterase (AChE) inhibition activity. The dichloromethane (DCM) and methanol (MeOH) fractions exhibited dose-dependent acetylcholinesterase inhibitory activity. The highest AChE inhibitory activities were from leaves (DCM fraction, IC50 = 0.1053 mg ml-1) and young stems (MeOH fraction, IC50 = 0.478 mg ml-1). High inhibitory activity against cyclooxygenase (COX-1 and COX-2) enzymes was observed. All extracts and fractions showed high COX-1 enzyme inhibition (90.7-100%). Petroleum ether (PE) and dichloromethane fractions also exhibited high inhibition against COX-2 enzyme (77.7-92.6%). The pharmacological activities observed suggest that S. birrea renewable plant parts (leaves and young stems) provide a substantial source of medicinal secondary metabolites. Based on these results, plant part substitution can be a practical conservation strategy for this species

Applications of cytokinins in horticultural fruit crops : trends and future prospects

CITATION: Aremu, Adeyemi O. et al. 2020. Applications of cytokinins in horticultural fruit crops : trends and future prospects. Biomolecules, 10(9):1222, doi:10.3390/biom10091222.The original publication is available at: https://mdpi-res.comCytokinins (CKs) are a chemically diverse class of plant growth regulators, exhibiting wide-ranging actions on plant growth and development, hence their exploitation in agriculture for crop improvement and management. Their coordinated regulatory effects and cross-talk interactions with other phytohormones and signaling networks are highly sophisticated, eliciting and controlling varied biological processes at the cellular to organismal levels. In this review, we briefly introduce the mode of action and general molecular biological effects of naturally occurring CKs before highlighting the great variability in the response of fruit crops to CK-based innovations. We present a comprehensive compilation of research linked to the application of CKs in non-model crop species in different phases of fruit production and management. By doing so, it is clear that the effects of CKs on fruit set, development, maturation, and ripening are not necessarily generic, even for cultivars within the same species, illustrating the magnitude of yet unknown intricate biochemical and genetic mechanisms regulating these processes in different fruit crops. Current approaches using genomic-to-metabolomic analysis are providing new insights into the in planta mechanisms of CKs, pinpointing the underlying CK-derived actions that may serve as potential targets for improving crop-specific traits and the development of new solutions for the preharvest and postharvest management of fruit crops. Where information is available, CK molecular biology is discussed in the context of its present and future implications in the applications of CKs to fruits of horticultural significance.Publisher's versio

An overview on Leucosidea sericea Eckl. & Zeyh.: A multi-purpose tree with potential as a phytomedicine

Ethnopharmacological relevance: Leucosidea sericea (the sole species in this genus) is a tree species found in southern Africa and possesses several therapeutical effects against infectious diseases in humans and livestock. This review aims to document and summarize the botany, phytochemical and biological properties of Leucosidea sericea. Materials and methods: Using the term ‘Leucosidea sericea’, we systematically searched literature including library catalogues, academic dissertations and databases such as PubMed, SciFinder, Web of Science, Google Scholar and Wanfang. Taxonomy of the species was validated using ‘The Plant List’ (www.theplantlist.org). Results: Leucosidea sericea remains a widely used species among the different ethnic groups in southern Africa. The species is a rich source of approximately 50 essential oils and different classes of phytochemicals (phenolics, phloroglucinols, cholestane triterpenoids, alkaloids and saponins) which may account for their diverse biological properties. Documented biological activities which were mainly observed under in vitro systems included antimicrobial, anti-parasitic, antioxidant, acetylcholinesterase (AChE) inhibition and antiinflammatory properties. Preliminary safety tests on Leucosidea sericea extracts suggest moderate cytotoxic effects based on a few cell lines that were investigated. Conclusions: Leucosidea sericea possesses diverse medicinal potential with the antimicrobial, anti-parasitic, antioxidant and anti-inflammatory activities being the most prominent. The relative abundance and tendency of Leucosidea sericea to evade in nature suggest an abundant reservoir of raw materials for potential commercialization (upon validation of its pharmacological potential). However, more stringent investigations on the extracts (and isolated bioactive compounds) focusing on the mode of actions, which will inevitably unravel their pharmacokinetics, pharmacodynamics and clinical relevance remain pertinent.University of KwaZulu-Natal (Pietermaritzburg)Cape Peninsula University of Technology (Cape Town)National Research Foundation (Grant number 98028 and 109508), South Afric

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

Abstract Background Medicinal 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. Methods The 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. Results The 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. Conclusions The 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.</p

2007. Determinants of farmer demand for fee-forservice extension in Zimbabwe‘s agricultural sector: the case

Abstract The increasing complexity of African agriculture has put greater pressure on agricultural education and extension. The important role played by agricultural extension has led man

2007. Determinants of farmer demand for fee-forservice extension in Zimbabwe‘s agricultural sector: the case

Abstract The increasing complexity of African agriculture has put greater pressure on agricultural education and extension. The important role played by agricultural extension has led man

Growth-promoting effects of a seaweed concentrate at various pH and water hardness conditions

Kelpak® – a liquid seaweed concentrate made from the kelp Ecklonia maxima (Osbeck) Papenfuss – is used as a natural biostimulant to promote rooting and improve yield in crops. Plant–soil environmental conditions and the chemistry of water used for irrigation may affect the efficiency of Kelpak. The effect of pH (pH 4.5, 6.5 and 8.5) and water hardness (200 mg/L and 400 mg/L Ca2+) on the growth-promoting ability of Kelpak was assessed using the mungbean rooting bioassay and in a pot trial with Swiss chard. Kelpak promoted rooting in all the treatments in the mungbean bioassay with maximum rooting generally achieved with 20% Kelpak. With 20% Kelpak, the addition of 200 mg/L and 400 mg/L Ca2+ decreased rooting at pH 4.5, increased rooting at pH 6.5 and did not affect rooting at pH 8.5. A similar trend was observed in the pot trial with Swiss chard: leaf and root (fresh weight) and pigment content (chl a, chl b and carotenoids) improved with the addition of 200 mg/L Ca2+ + 5% Kelpak at pH 6.5 or pH 8.5, while Kelpak was able to partially mask the negative effect of 200 mg/L Ca2+ at pH 4.5. These results suggest that while Kelpak is most effective in neutral pHs, it can be used to promote plant growth in a wide range of pH and water hardness conditions

Determination of Mineral Constituents, Phytochemicals and Antioxidant Qualities of Cleome gynandra, Compared to Brassica oleracea and Beta vulgaris

The study compared mineral, chemical and antioxidant qualities of Cleome gynandra, a wild leafy vegetable, with two widely consumed commercial vegetables, Brassica oleracea and Beta vulgaris. Mineral nutrients were quantified with inductively coupled plasma mass spectrometry (ICP-MS), phenolic compounds using ultra-high performance liquid chromatography coupled to a mass spectrometer (UHPLC-MS) and β-carotene and vitamin C using high performance liquid chromatography with a photodiode array detector (HPLC-PDA). The antioxidant potential was evaluated using 2,2–diphenyl−1–picryl hydrazyl (DPPH) and oxygen radical absorbance capacity (ORAC) assays. Cleome gynandra had highest concentrations of phosphorus, potassium, calcium, iron, zinc, ascorbic acid, total phenolics, and flavonoids; whereas sodium, magnesium, manganese, copper and β-carotene were higher for B. vulgaris. The significantly higher antioxidant activity (P ≤ 0.05) exhibited by C. gynandra in comparison to the two commercial vegetables may be due to its significantly high levels of vitamin C and phenolic acids. These findings on the mineral, chemical and antioxidant properties of C. gynandra provide compelling scientific evidence of its potential in adding diversity to the diet and contributing toward the daily nutritional requirements of millions of people for food and nutritional security

Natural Antioxidants: Fascinating or Mythical Biomolecules?

Research on the use, properties, characteristics and sources of antioxidants especially phenolic compounds, flavonoids, vitamins, synthetic chemicals and some micronutrients began in the late 18th century. Since then antioxidant research has received considerable attention and over a hundred thousand papers have been published on the subject. This has led to a rampant use of antioxidants in order to try to obtain and preserve optimal health. A number of nutraceuticals and food supplements are frequently fortified with synthetic or natural antioxidants. However, some research outcomes have led to the belief that antioxidants exist as mythical biomolecules. This review provides a critical evaluation of some common in vitro antioxidant capacity methods, and a discussion on the role and controversies surrounding non-enzymatic biomolecules, in particular phenolic compounds and non-phenolic compounds, in oxidative processes in an attempt of stemming the tidal wave that is threatening to swamp the concept of natural antioxidants