Academic literature on the topic 'Agathosma betulina'

Agathosma betulina, commonly known as buchu, has been used for centuries by the indigenous people of South Africa for medicinal purposes. Currently, the essential oils from buchu are used in medicine, food flavorings, and aromatic oils. Increased exploitation of natural growing buchu in the Fynbos biome and a worldwide shortage of buchu oil encouraged commercial cultivation in South Africa. The root-knot nematode (Meloidogyne spp.) is one of the most common plant-parasitic nematodes found on commercial crops grown in the Western Cape. It has also been isolated from the soil and roots of plants in the natural Fynbos vegetation (2). In June 2003, a nursery propagating buchu plants experienced problems with poor growth. Examination of the buchu roots under a stereo microscope showed extensive galling with large numbers of female root-knot nematodes with eggsacs. Nematode extractions of the soil were also done. Only second-stage juveniles of Meloidogyne spp. (311 per 250 ml of soil) were recovered. A polymerase chain reaction (PCR)-based diagnostic method (1) was used for the identification of the root-knot nematode species. Ten intact females were dissected from the roots and individually placed directly in 5 μl drops of 1× PCR reaction buffer (16 mM [NH4]2SO4, 67 mM tris-HCL, pH 8.8, 0.1% vol/vol Tween 20) ontaining 60 μg/ml of proteinase K. The tube was kept at -80°C for a minimum of 10 min. The tube was incubated at 60°C for 15 min and 5 min at 95°C. The PCR amplifications were then prepared directly in the same tube. Amplified DNA fragments were digested with HinfI and DraI. The digested DNA was loaded on a 2% agarose gel, separated by electrophoresis, and detected by ethidium bromide staining. The digested amplified DNA fragments correspond to those of Meloidogyne javanica. Morphological characteristics were used to verify the PCR-based identification of the nematode. To our knowledge, this is the first report of M. javanica causing extensive galling on the roots of Agathosma betulina. Visual damage to the roots indicates the root-knot nematode to be an important threat to the commercial cultivation of buchu. References: (1) R. Knoetze. Potential of the polymerase chain reaction for the identification of plant-parasitic nematodes. M.Sc. thesis. University of Stellenbosch, Stellenbosch, South Africa, 1999. (2) A. J. Meyer, S. Afr. J. Enol. Vitic., 20:75, 1999.

South Africa has offered the world two indigenous aromatic plants from which commercially important natural products have been developed: Pelargonium graveolens (and its hybrids) the source of geranium oil and Agathosma betulina, from which ‘Buchu’ oil is produced. Despite the historical use of ‘Buchu’ and the commercial interest developed around this coveted indigenous resource the (limited) research has not been coherently assembled. This overview aims to unite aspects on the botany, traditional and modern day uses, chemistry and pharmacological data on ‘Buchu’ which is undeniably one of South Africa’s most renowned botanical assets.

Increased management through domestication is the predicted, and often necessary, commercialisation outcome of a wild resource which is subject to a demanding market that promotes competition amongst producers and the depletion of wild stocks. This has been the case for commercial buchu (Agathosma betulina and Agathosma crenulata), a historically wild collected plant which has been cultivated on a large-scale in selected areas of the Western Cape Province of South Africa. Buchu is an endemic, aromatic shrub around which a lucrative industry spanning diverse and distant markets has developed. Alongside its medicinal properties, buchu is primarily valued for its essential oil which is exported for use in international flavour and fragrant industries. The aim of this study was to conduct an overview of the local buchu industry with a focus on how cultivation has impacted on the general trade, the different actors involved and the conservation of the plant. A shift in buchu production to large-scale, agricultural enterprises raises certain questions for the involvement of rural harvesters in the trade, especially with regard to their inclusion and the sharing of benefits arising from commercialisation. Accordingly, this research sought to identify the social and economic impacts of buchu cultivation while also exploring the environmental impacts associated with large-scale farming of the plant. The methods employed in this research were primarily qualitative, based on semi-structured interviews conducted with key actors involved in the buchu trade, including harvesters, farmers, industry representatives and environmental authorities. The study revealed that while the harvesting of buchu is an important economic activity for harvester communities, the cultivation of buchu has played a limited role in local livelihoods with cultivation mainly being confined to large-scale, commercial operations in the hands of wealthy farmers and private processing companies. The findings of this research also shed light on the shortcomings of national access and benefit-sharing legislation which has failed to secure commercial benefits for the rural poor involved in the trade. From an environmental perspective, the cultivation of buchu has contributed to the conservation of the plant in the wild through offsetting harvesting pressures experienced by wild populations, but has also contributed to the destruction of naturally occurring vegetation.

Abstract Few in vitro screening assays for biological activities of plant extracts consider the potential effect of the gastrointestinal system on orally consumed plant extracts. Crude water and methanol extracts of Tarchonanthus camphoratus (wild camphor) and Agathosma betulina (‘buchu’) were prepared and exposed to simulated gastric fluid and simulated intestinal fluid during dissolution studies to address this aspect. The crude extracts and resulting simulated gastric fluid and simulated intestinal fluid products were screened for antimicrobial activity against Staphylococcus aureus (ATCC 25923), Enterococcus faecalis (ATCC 29212), Escherichia coli (ATCC 25922) and Proteus vulgaris (ATCC 33420). The T. camphoratus crude extract exhibited antimicrobial activity which was reduced after exposure to simulated gastric fluid. After exposure to simulated intestinal fluid no antimicrobial activity was detected, which suggests chemical alteration or degradation of the active compounds. For A. betulina, the crude water extract and simulated gastric fluid product exhibited no antimicrobial activity, while the simulated intestinal fluid product exhibited antimicrobial activity. This suggests activation of antimicrobial constituents during exposure to simulated intestinal fluid. The chemical composition profiles of the crude extracts and products were determined by means of liquid chromatography coupled to an ultraviolet detector (LC-UV) and a mass spectrometer (LC-MS) to qualitatively assess the effect of exposure to simulated gastrointestinal conditions on the chemical composition of the extracts. In many cases, the peak area of compounds decreased after exposure to simulated gastric fluid and simulated intestinal fluid, while the peak area of other compounds increased. Thus, it can be deduced that the antimicrobial activity and chemical composition was altered after exposure to intestinal conditions during dissolution studies.

Thesis submitted in fulfilment of the requirements for the degree Master of Technology: Horticultural Sciences in the Faculty of Applied Sciences at the CAPE PENINSULA UNIVERSITY OF TECHNOLOGY Supervisor: Dr L Kambizi Co-supervisor: Dr NP Makunga Cape Town December 2013
Agathosma betulina (Berg.) Pillans, previously known as Barosma betulina, is a member of the Rutaceae family, and indigenous to the fynbos botanical biome of the Western Cape of South Africa. It is commonly known as buchu. Extracts as well as powdered leaves have traditionally been used for the treatment of various ailments. The increase in the international demand for A. betulina for health as well as food and beverage benefits, have raised concerns over exploitation of wild populations and the lack of horticultural information necessitates this study to evaluate the propagation of this economical important species. The main objective of this study was to establish a simple and highly productive micropropagation protocol for A. betulina through experimenting with nodal explants. Testing of the effect of various treatments (physical scarification, chemical scarification, GA, stratification, smoke and combinations thereof) on the in vitro germination of A. betulina seeds was done to elucidate the factors which control seed germination. The study revealed that the physical scarification and smoke-induced germination had a significant effect on germination percentages. In terms of germination rate, the radical generally started to appear after approximately 10 days in the physical scarification with smoke treatment. Initial decontamination methods with the exposure of various concentrations of NaOCl gave fatal results, however 1.5% NaOCl had more phenolic reactions rather than fungal or bacterial contamination. Interestingly, contamination rates of explants were influenced by the stage of maturity of the explant material. This plant material was used to test different strengths of regeneration media, to ensure that the explants receive ample nutrients. Results made exhibited that ½ MS was the best strength for growing A. betulina nodal explants. Compared comparison between in vitro derived explants and ex vitro collected explants showed that the ex vitro derived explants had significant results, but the explants lost vigour soon after the initial exponential growth leading to the explants dying off. Furthermore, ex vitro decontaminated plant material was not economically viable to continue with. Seedlings derived from germinated seeds appeared to be the preferred method of propagation as this spent the least time in culture and produced a stable plant with an established root system, which is essential during the hardening off process after in vitro growth. When exposing nodal explants to phytohormone 2,4-D it responds best to dosages 0.5mg Lˉ¹ and 1mg Lˉ¹. Phytohormone BA was very effective in producing soft friable callus. The best results were shown when 0.5mg Lˉ¹ BA was applied to ½ MS media. For both shoot length and multiple shoot production, a combination of phytohormones BA-NAA (1: 0.5mgLˉ¹) had the most significant results. Interestingly, a higher phytohormone concentration of NAA is necessary to develop multiple adventitious roots. The effect of 3mg Lˉ¹ was significant in that it resulted in multiple adventitious roots, but fewer calli was observed in this treatment. Micropropagation becomes valuable as little attention between subcultures is needed; making it less labour intensive compared to conventional nursery propagation systems where weeding watering and spraying of plants are labour intensive. In the traditional world of medicine, more so in Southern Africa, extracts are prepared by adding boiling water to the plant material; however commercial ethanol is used as an extractant. Establishment of the essential oil quality of the in vitro cultures post exposure to various treatments was done. Analysis of essential oils from A. betulina resulted in the identification of twenty one compounds. The results showed qualitative as well as quantitative differences amongst the samples used in the study. The highest relative concentration of limonene was observed in the callus of nodal explants after it was exposed to 0.5mg lˉ¹ NAA. No pulegone was found in this treatment making it ideal for limonene production. This suggests that liquid culture with the same treatment may produce more calli making it ideal for the production of limonene.

The phytochemistry, antimicrobial and antioxidant activity of several extracts of the South African traditional herbal medicine Agathosma betulina (buchu) was investigated. Antifungal screening showed that the reputed active principle in buchu oil, diosphenol, is not the main active component in the herb whereas additional compounds such as hydroxycinnamic acids and lipids may be contributory. However antibacterial screening showed that all the extracts showed very low activity against micro organisms responsible for the pathogenesis of the urinary tract. At a concentration of 1 mg.ml." acetone, 80% MeOH and aqueous extracts exhibited 75% to >90% activity against the stable radical DPPHe whereas the essential oil was weakly active. Furthermore a poor correlation between the total phenolic content and radical scavenging activity of the extracts was observed. The only extract that suppressed the oxidation of linoleic acid was the acetone extract. Four lipophilic flavonoids were detected in the acetone extract and were tentatively identified as the 3 and 3,3'-dimethyl ethers of quercetin and the 3 and 3,4' dimethyl ethers of kaempferol. Three antiradical flavonol glycosides 3, 4 and 5 were detected in the 80% MeOH extract. Flavonoid 5 was identified as rutin and this compound was the main antiradical component in the aqueous extracts. Flavonoid 3 was hyperoside whereas flavonoid 4 was identified as a novel compound designated agathosin on the basis of spectroscopic evidence. Agathosin was shown to be quercetin-3-0-p-o-glucoside esterified with oleuropeic acid, a monoterpenoid carboxylic acid. A rational basis for its structure and the significance of agathosin in the light of the traditional medicinal use of buchu is discussed in addition to its potential biological activity. Furthermore the results of the antimicrobial, antioxidant activity and total phenolic content of the plant extracts are discussed in the light of their chemical composition. In summation this work showed that buchu does not have a direct antimicrobial action, highlighted the importance of using more than one type of assay when assigning antioxidant activity to herbs and their extracts and showed that by assessing extracts from the same plant for different typesof bioactivity novel compounds may be detected.

Thesis (MSc)--University of Stellenbosch, 2005.
ENGLISH ABSTRACT: The Mountain Fynbos biome, a division of the Cape Floristic Region (CFR), is home to round-leafed Buchu [Agathosma betulina (Berg.) Pillans], one of South Africa’s best-known endangered herbal medicinal plants. Agathosma betulina is renowned as a traditional additive to brandy or tea, which is used for the treatment of a myriad of ailments. In its natural habitat, A. betulina thrives on mountain slopes in acid and highly leached gravelly soils, with a low base saturation and low concentrations of organic matter. To adapt to such adverse conditions, these plants have formed mutualistic symbioses with arbuscular mycorrhizal (AM) fungi. In this study, the effect of indigenous AM taxa on the physiology of A. betulina is investigated. In addition, the AM taxa responsible for these physiological responses in the plant were identified using morphological and molecular techniques. Agathosma betulina was grown under glasshouse conditions in its native rhizosphere soil containing a mixed population of AM fungi. Control plants, grown in the absence of AM fungi, were included in the experimentation. In a time-course study, relative growth rate (RGR), phosphorus (P)-uptake, P utilization cost, and carbon (C)-economy of the AM symbiosis were calculated. The data showed that the initial stages of growth were characterized by a progressive increase in AM colonization. This resulted in an enhanced P-uptake in relation to non-AM plants once the symbiosis was established. Consequently, the lower P utilization cost in AM plants indicated that these plants were more efficient in acquiring P than non-AM plants. When colonization levels peaked, AM plants had consistently higher growth respiration. This indicated that the symbiosis was resulting in a C-cost to the host plant, characterized by a lower RGR in AM plants compared to non-AM plants. Arbuscular mycorrhizal colonization decreased with increasing plant age that coincided with a decline in P-uptake and growth respiration, along with increases in RGR to a level equal to non-AM plants. Consequently, the AM benefit was only observed during the initial stages of growth. In order to identify the AM fungi in planta, morphological and molecular techniques were employed, which indicated colonization by AM fungi belonging to the genera Acaulospora and Glomus. Phylogenetic analyses of a dataset containing aligned 5.8S ribosomal RNA gene sequences from all families within the Glomeromycota, including sequences obtained during the study, supported the above mentioned identification.
AFRIKAANSE OPSOMMING: Die Fynbos bergbioom, ‘n onderafdeling van die Kaapse Floristiese Streek, huisves rondeblaar Boegoe [Agathosma betulina (Berg.) Pillans], een van Suid Afrika se bekendste bedreigde medisinale plante. Agathosma betulina is bekend vir sy gebruik as tinktuur vir die behandeling van verskeie kwale. Die plant kom voor in bergagtige streke, in suur en mineraal-arm grond, met ‘n lae organiese inhoud. Gevolglik, om aan te pas by hierdie ongunstige kondisies, vorm die plante simbiotiese assosiasies met blaasagtige, struikvormige mikorrisa (BSM). In die huidige studie is die effek van hierdie BSM op die fisiologie van A. betulina ondersoek. Die identiteit van die BSM is ook gevolglik met morfologiese en molekulêre identifikasie tegnieke bepaal. Agathosma betulina plante is onder glashuis kondisies in hul natuurlike grond gekweek, wat ‘n natuurlike populasie van BSM bevat het. Kontroles is ook in die eksperiment ingesluit en hierdie stel plante is met geen BSM geïnokuleer nie. Gevolglik is die relatiewe groeitempo, fosfor opname, fosfor verbuikerskoste asook die koolstof ekonomie van die plante bereken. Die data het getoon dat die eerste groeifase gekarakteriseer is deur toenames in BSM kolonisasie vlakke. Dit het tot ‘n hoër fosfor opname in BSM geïnokuleerde plante gelei. Die laer fosfor verbuikerskoste gedurende hierdie fase het aangedui dat die plante wat geïnokuleer is met BSM oor beter meganismes beskik het om fosfor uit die grond te bekom. Toe BSM kolonisasie vlakke gepiek het, was groei respirasie hoër in BSM geïnokuleerde plante as in die kontroles. Dit het aangedui dat die BSM kolonisasie van plante tot hoër koolstof kostes vir hierdie plante gelei het, wat weerspieël is in die laer groeitempo van die BSM geïnokuleerde plante. Die BSM kolonisasie vlakke het gedaal met toenemende ouderdom van hul gasheer plante, wat gekarakteriseer is deur ‘n laer opname van fosfor en laer groei respirasie, tesame met ‘n toename in relatiewe groeitempo tot vlakke soortgelyk aan die van die kontrole plante. Die BSM voordele vir die plant is dus net gedurende die eerste groeifase waargeneem. Die BSM wat verantwoordelik is vir hierdie fisiologiese veranderinge is gevolglik geïdentifiseer met behulp van morfologiese en molekulêre tegnieke en dit is gevind dat BSM wat behoort tot die genera Acaulospora en Glomus binne hierdie plante voorkom. Filogenetiese analise gegrond op opgelynde 5.8S ribosomale RNA geen volgordes afkomstig van al die families binne Glomeromycota asook volgordes gevind in die studie, het die bogenoemde identifikasie gestaaf.

Thesis (PhD (Microbiology))--University of Stellenbosch, 2010.
Dissertation presented for the degree of Doctor of Philosophy in Microbiology at the University of Stellenbosch.
ENGLISH ABSTRACT: The interaction between a soil yeast, Cryptococcus laurentii and a medicinal plant, Agathosma betulina (Berg.) Pillans (Rutaceae), was studied. Cryptococcus laurentii CAB 578 was isolated from the rhizosphere of wild A. betulina and liquid chromatographytandem mass spectrometry analysis revealed that the yeast was capable of producing polyamines such as cadaverine and spermine. Since the exogenous application of polyamines are known to impact on root growth, these findings supported the results obtained when A. betulina seedlings grown under axenic and low nutrient conditions were inoculated with C. laurentii CAB 578 and cultivated for five months under glasshouse conditions. The presence of the yeast increased root growth by 51%. Using soil dilution plates, it was demonstrated that yeast numbers were greater in the vicinity of the roots than in the bulk soil. Furthermore, fluoromicroscopy, in combination with the fluorescent probes Calcofluor White and Fungolight revealed the presence of metabolic active yeast colonies on the rhizoplane. The first part of the study thus provided evidence for a symbiosis between A. betulina and C. laurentii CAB 578. During the second part of the investigation, the effect of this symbiosis on quantitative elemental distribution in A. betulina roots grown under axenic, nutrient-poor conditions was assessed using micro-particle-induced x-ray emission spectrometry. To aid in the interpretation of heterogeneous elemental distribution patterns, apoplastic barriers and endophytic C. laurentii CAB 578 in root tissues were located using fluorescence microscopy and transmission electron microscopy, respectively. The results showed that the average concentrations of iron, manganese and phosphorus were significantly (P < 0.05) higher within roots of yeast-inoculated plants, compared to control plants. It was shown that the yeast was not a root endophyte and that elemental enrichment in the epi/exodermal-outer cortical tissues correlated with the presence of Casparian bands in the exodermal cells of both treatments. This was the first report describing the role of a soil yeast as a plant nutrient-scavenging microsymbiont. In the final part of the investigation, the effect of C. laurentii CAB 578 on the photosynthetic nitrogen, phosphorus and water-use efficiencies, as well as the carbon economy of A. betulina was studied. Agathosma betulina plants inoculated with C. laurentii CAB 578, as well as controls, were grown under axenic conditions and the following parameters measured: Apparent photon yield, foliar nitrogen and phosphorus concentrations, leaf dark respiration, maximum photosynthetic rate, photosynthetic nitrogen-use efficiency, photosynthetic phosphorus-use efficiency, photosynthetic wateruse efficiency, root construction cost, stomatal conductance, substomatal CO2 and transpiration rate. The data showed that the higher photosynthetic resource-use efficiencies in yeast-inoculated plants were a consequence of higher maximum rates of CO2 assimilation, which was not related to foliar nitrogen and phosphorus content. We hypothesize that photosynthetic stimulation in yeast-inoculated plants was a result of the increased demand for photosynthates of the yeast-root symbiosis. In summary, the study revealed that a symbiosis exists between A. betulina and the soil yeast C. laurentii CAB 578. This interaction has a significant effect on the size of the yeast population as well as on the physiology of the plant.
AFRIKAANSE OPSOMMING: Die interaksie tussen ‘n grondgis, Cryptococcus laurentii, en ‘n medisinale plant, Agathosma betulina (Berg.) Pillans, is ondersoek. Cryptococcus laurentii CAB 578 is vanuit die risosfeer van A. betulina in sy natuurlike omgewing geisoleer en vloeistof chromatografie tandem massa spektrofotometriese analise het bewys dat die gis poliamiene insluitend kadaverien en spermien produseer. Dit is bevind dat die eksogene aanwending van poli-amiene wortelgroei bevorder. Hierdie bevinding staaf die waargenome 51% verhoging in wortelgroei van mikroob-vrye A. betulina saailinge geinokuleer met C. laurentii CAB 578 en gekweek vir vyf maande onder lae nutriënt kondisies in ‘n glashuis. Met gebruik van die grond verdunningsplaat-metode, is dit verder bewys dat gisgetalle hoër was in die teenwoordigheid van wortels as in die omliggende grond. Dit is ook bewys met die gebruik van die fluoressente peilers Calcofluor White en Fungolight, in kombinasie met fluoressensie-mikroskopie, dat metabolies aktiewe giste die wortels se oppervlak gekoloniseer het. Die eerste deel van die studie het dus bewys dat ‘n simbiose tussen A. betulina en C. laurentii CAB 578 bestaan. Tydens die tweede deel van die ondersoek is die effek van C. laurentii CAB 578 op die konsentrasie en verspreiding van elemente binne A. betulina wortels, gekweek onder lae-nutriënt, mikroob-vrye kondisies, bepaal met behulp van mikro-partikel geinduseerde X-straal emissie spektrofotometrie. Om die interpretasie van heterogene verspreidingspatrone van die onderskeie elemente te ondersteun, is daar met behulp van fluoressensie en transmissie-elektron-mikroskopie vir apoplastiese versperrings en endofitiese C. laurentii CAB 578 in die wortelweefsel getoets. Dit is bevind dat die gemiddelde konsentrasie van fosfaat, mangaan en yster beduidend (P < 0.05) hoër was in gis-geinokuleerde plante, as in kontrole plante. Die gis was nie ‘n wortel endofiet nie en elementale verryking in die epi/eksodermale-buitenste korteks weefsels het gekorreleer met Casparian bande in die eksodermale selle van beide behandelings. Hierdie was die eerste verslag wat die rol van ‘n grondgis as ‘n nutriënt-bekommende mikrosimbiont vir plante beskryf het. In die laaste gedeelte van hierdie ondersoek is die effek van C. laurentii CAB 578 op die fotosintetiese fosfaat, stikstof en water-verbruiksdoeltreffendheid, asook die koolstof ekonomie in mikroob-vrye Agathosma betulina plante geinokuleer met C. laurentii CAB 578 asook kontrole plante bestudeer. Die volgende parameters is getoets: Blaar donker respirasie, blaar fosfaat en stikstof konsentrasies, fotosintetiese fosfaatverbruiksdoeltreffendheid, fotosintetiese stikstof-verbruiksdoeltreffendheid, fotosintetiese water-verbruiksdoeltreffendheid, huidmond konduktansie, huidmond CO2 konsentrasie, klaarblyklike foton opbrengs, maksimum fotosintetiese spoed, wortel konstruksie-koste, en transpirasie spoed. Die resultate het getoon dat die hoër maksimum fotosintestiese spoed in gis-geinokuleerde plante gelei het tot ‘n hoër fotosintetiese verbruiksdoeltreffendheid van fosfaat, stikstof en water en dat dit nie verband gehou het met blaar fosfaat en stikstof konsentrasies nie. Dit word voorgestel dat die stimulasie van fotosintese in gisgeinokuleerde plante ‘n gevolg is van die verhoogde aanvraag na fotosintaat deur die giswortel simbiose. Om op te som, die bevindings van hierdie studie het bewys dat ‘n simbiose tussen A. betulina en C. laurentii CAB 578 bestaan. Hierdie simbiose het ‘n beduidende effek op die populasie grootte van die gis sowel as die fisiologie van die plant.

Magister Scientiae Dentium - MSc(Dent)
Candida albicans is a clinical fungal isolate that is most frequently isolated from different host niches, and is implicated in the pathogenesis of several fungal infections, including oral candidiasis. The pathogenesis and antifungal resistance mechanisms of Candida species are complex and involve several pathways and genes. Oral candidiasis incidence rates are rapidly increasing, and the increase in resistance to conventional antifungals has led to the need to develop innocuous and more efficacious treatment modalities. The purpose of this study was to explore a single pot process for phytosynthesis of zinc oxide nanoparticles (GZnO NPs) and to assess their antifungal potential.

The aim of this thesis was to explore the socio-economic factors that impact on the sustainable harvesting of buchu in the Western Cape of South Africa. Some of the factors that were explored include poverty, natural resource tenure, legislation, and local practices with regard to the harvesting of buchu.