Nitrogen application influences quality, pharmacological activities and metabolite profiles of Athrixia phylicoides DC. (Bush tea) cultivated under greenhouse and field conditions

Bush tea (Athrixia phylicoides DC.) is a popular aromatic plant, indigenous to South Africa. For many decades, indigenous people have widely consumed the tea as a healthy beverage and a medicine. The objective of the study was to investigate the influence of nitrogen fertiliser application on the quality, pharmacological activities and metabolite profiles of the leaves and twigs of bush tea cultivated under greenhouse and field conditions. Treatments consisted of 0, 75, 150, 225, 300 and 375 kg/ha N, arranged in a randomised complete block design with five replicates. Parameters measured were leaf tissue nitrogen, total polyphenols, total flavonoids, total tannins, total antioxidants, antimicrobial activity and cytotoxicity. In addition, metabolites were identified and measured using gas chromatography linked to mass spectrometry (GC-MS) and liquid chromatography linked to mass spectrometry (LC-MS). The results of this study showed a significant quadratic response of total polyphenols, total flavonoids, total tannins and total antioxidants of bush tea leaves and twigs, irrespective of the growth environment ranging from 150 to 225 kg/ha N. The results further suggested that, despite the growing environment, nitrogen treatments did not improve the minimum inhibition concentration and minimum microbicide concentration of bush tea with extract concentrations ranging from 25.0 to 3.1 mg/g. Bush tea was found to be non-toxic (IC50 above 100) under both growing conditions, irrespective of nitrogen treatments. Despite trials being conducted under greenhouse and field conditions and with different nitrogen treatments, there were no distinct differences regarding the gas chromatography linked to mass spectrometry and liquid chromatography linked to mass spectrometry compounds. The future aims of the study are to investigate the effects of nitrogen fertilisation, the timing of nitrogen fertilisation and the planting density on the yield, quality, plant metabolites and pharmacological activity of field-grown bush tea

Resprouters versus reseeders : are wild rooibos ecotypes genetically distinct?

CITATION: Brooks, J. et al. 2021. Resprouters versus reseeders : are wild rooibos ecotypes genetically distinct? Frontiers in Genetics, 12:761988, doi:10.3389/fgene.2021.761988.The original publication is available at https://www.frontiersin.orgAspalathus linearis (Burm. F.) R. Dahlgren (Fabaceae) or rooibos, is a strict endemic species, limited to areas of the Cederberg (Western Cape) and the southern Bokkeveld plateau (Northern Cape) in the greater Cape Floristic Region (CFR) of South Africa. Wild rooibos, unlike the cultivated type, is variable in morphology, biochemistry, ecology and genetics, and these ecotypes are broadly distinguished into two main groups, namely, reseeders and resprouters, based on their fire-survival strategy. No previous assessment of genetic diversity or population structure using microsatellite markers has been conducted in A. linearis. This study aimed to test the hypothesis that wild rooibos ecotypes are distinct in genetic variability and that the ecotypes found in the Northern Cape are differentiated from those in the Cederberg that may be linked to a fire-survival strategy as well as distinct morphological and phytochemical differences. A phylogeographical and population genetic analyses of both chloroplast (trnLF intergenic region) and newly developed species-specific nuclear markers (microsatellites) was performed on six geographically representative wild rooibos populations. From the diversity indices, it was evident that the wild rooibos populations have low-to-moderate genetic diversity (He: 0.618–0.723; Ho: 0.528–0.704). The Jamaka population (Cederberg, Western Cape) had the lowest haplotype diversity (H = 0.286), and the lowest nucleotide diversity (π = 0.006) even though the data revealed large variations in haplotype diversity (h = 0.286–0.900) and nucleotide diversity (π = 0.006–0.025) between populations and amongst regions where wild rooibos populations are found. Our data suggests that populations of rooibos become less diverse from the Melkkraal population (Suid Bokkeveld, Northern Cape) down towards the Cederberg (Western Cape) populations, possibly indicative of clinal variation. The largest genetic differentiation was between Heuningvlei (Cederberg, Western Cape) and Jamaka (FST = 0.101) localities within the Cederberg mountainous region, and, Blomfontein (Northern Cape) and Jamaka (Cederberg) (FST = 0.101). There was also a significant isolation by distance (R2 = 0.296, p = 0.044). The presence of three main clusters is also clearly reflected in the discriminant analysis of principal components (DAPC) based on the microsatellite marker analyses. The correct and appropriate management of wild genetic resources of the species is urgently needed, considering that the wild Cederberg populations are genetically distinct from the wild Northern Cape plants and are delineated in accordance with ecological functional traits of reseeding or resprouting, respectively. The haplotype divergence of the ecotypes has also provided insights into the genetic history of these populations and highlighted the need for the establishment of appropriate conservation strategies for the protection of wild ecotypes.Publisher's versio

Fusarium inhibition by wild populations of the medicinal plant Salvia africana-lutea L. linked to metabolomic profiling

Publication of this article was funded by the Stellenbosch University Open Access Fund.The original publication is available at http://www.biomedcentral.com/1472-6882/14/99Abstract Background: Salvia africana-lutea L., an important medicinal sage used in the Western Cape (South Africa), can be termed a ‘broad-spectrum remedy’ suggesting the presence of a multiplicity of bioactive metabolites. This study aimed at assessing wild S. africana-lutea populations for chemotypic variation and anti-Fusarium properties. Methods: Samples were collected from four wild growing population sites (Yzerfontein, Silwerstroomstrand, Koeberg and Brackenfell) and one garden growing location in Stellenbosch. Their antifungal activities against Fusarium verticillioides (strains: MRC 826 and MRC 8267) and F. proliferatum (strains: MRC 6908 and MRC 7140) that are aggressive mycotoxigenic phytopathogens were compared using an in vitro microdilution assay. To correlate antifungal activity to chemical profiles, three techniques viz. Gas chromatography-mass spectrometry (GC-MS); Liquid chromatography-mass spectrometry (LC-MS) and 1H Nuclear Magnetic Resonance (NMR) were employed. Principal Component Analysis (PCA) was applied to the NMR data. The partial least squares-discriminant analysis (PLS-DA) was used to integrate LC-MS and NMR data sets. All statistics were performed with the SIMCA-P + 12.0 software. Results: The dichloromethane:methanol (1:1; v/v) extracts of the plant species collected from Stellenbosch demonstrated the strongest inhibition of F. verticillioides and F. proliferatum with minimum inhibitory concentration (MIC) values of 0.031 mg ml-1 and 0.063 mg ml-1 respectively. GC-MS showed four compounds which were unique to the Stellenbosch extracts. By integrating LC-MS and 1H NMR analyses, large chemotype differences leading to samples grouping by site when a multivariate analysis was performed, suggested strong plant-environment interactions as factors influencing metabolite composition. Signals distinguishing the Stellenbosch profile were in the aromatic part of the 1H NMR spectra. Conclusions: This study shows the potential of chemotypes of Salvia africana-lutea in controlling fungal growth and consequently mycotoxin production. Products for use in the agricultural sector may be developed from such chemotypes.Stellenbosch UniversityPublishers' Versio

In vitro propagation of the wild carrot Daucus carota L. subsp. halophilus (Brot.) A. Pujadas for conservation purposes

Daucus carota subsp. halophilus, is a wild crop relative of domestic carrot. It is an aromatic plant widely used in folk medicine due to recognized therapeutic properties of its essential oils. Experiments were carried out to evaluate the potential of in vitro propagation techniques to the conservation of this endemic and endangered taxon. The results showed that shoot tips of in vitro germinated seeds were able to proliferate in the presence of benzyladenine, with the best results being achieved using 4.4μM, both in the first and second cultures. Shoots rooted after being transferred to 1/2- Murashige and Skoog basal medium. The results indicated that the concentration of benzyladenine used during the multiplication phase did not interfere with the rate of root formation. The obtained plantlets were morphologically and anatomically identical to those obtained by seeds. Some of the in vitro produced shoots developed flowers that produced viable pollen. Plant regeneration was also achieved by somatic embryogenesis induction in cotyledons and root segments cultured in the presence of 4.5μM 2,4-dichlorophenoxyacetic acid. Somatic embryos converted into plantlets in a medium without growth regulators. Plants obtained either by shoot proliferation or somatic embryogenesis were acclimatized and are now growing at the Coimbra Botanical Garden. The first attempts to reintroduce these plants in the original habitat were successful. It can be concluded that the protocols developed are a useful approach to the conservation of this endemic species