Chicory (Cichorium intybus L) is a deep rooted biennial (or weakly perennial) composite that is grown as an annual in South Africa for its parsnip-like root. In its wild state the plant is a perennial with thin fibrous roots, but, through selection and propagation, cultivars with large, fleshy roots have been developed. From a purely horticultural standpoint, chicory is of interest as a pot-herb, a salad plant and as a root (Bailey, 1942) . As a root, the chicory plant represents an important article of commerce. In South Africa, the plants are grown virtually exclusively for their roots which, after drying, roasting and grinding, are used as an additive to, or substitute for, coffee (Introduction p. 11)

The biochemical changes taking place during soybean seed development and gennination, and some aspects of desiccation tolerance were assessed with reference to lipid peroxidation and antioxidant systems. During nonnal seed development, fresh weight and dry weight increased between 20 and 50 days after flowering (DAF), concomitant with the accumulation of triacylglycerols and sugar reserves, after which dry weight remained almost unchanged, and fresh weight decreased. Seed moisture content decreased rapidly during the last stages of development. High levels of lipid peroxidation were evident between 20 and 45 DAF, and decreased thereafter. An examination of antioxidant systems revealed that whereas total glutathione levels accumulated continuously throughout the 80 days of seed development, both dehydroascorbic acid (DHA) reductase and ascorbate free radical (AFR) reductase increased concurrently with the increase in total ascorbate content, and the overall levels did not decrease markedly during maturation drying. Ascorbate peroxidase (ASC POD) activity was high during the period ofgreatest ascorbate accumulation. Both catalase (CAn and superoxide dismutase (SOD) activities increased progressively during early seed development (20-40 DAF), but showed variable patterns of change during maturational drying, in marked contrast to ASC POD which declined from 40 DAF to undetectable levels at 70 DAF. An assessment of the relationship between the antioxidant systems and lipid peroxidation was made during imbibition and gennination, as it has been suggested that controlling free radicals was a critical event in early imbibition. Unexpectedly, lipid peroxidation increased progressively in both seeds and isolated axes, and were eight-fold higher at 48 hours of imbibition compared to dry tissues. A progressive, and co-ordinated, increase in CAT, total glutathione, total ascorbate pool, guaiacol POD, ASC POD, and SOD appeared to parallel the rise in lipid peroxidation in both whole seeds and axes. Variable responses were evident between seeds and axes for the enzymes AFR reductase and DHA reductase In order to gain a further insight into the dynamics of desiccation-tolerance and desiccationsensitivity, imbibing seeds were subjected to an unscheduled dehydration treatment, and then rehydrated for up to 24 hours. During these hydration-dehydration-rehydration (H-D-R) treatments, changes in lipid peroxidation and antioxidant systems were measured. Concurrent with the loss of viability in the axes of seeds dehydrated after 24 and 36 hours of imbibition, there were increases in both lipid peroxidation and solute leakage. Unscheduled drying was seen to be a critical stage, as intolerant axes showed four- to eightfold increases in lipid peroxidation, which were only partially reduced on subsequent rehydration. Tolerant axes, on the other hand, were able to maintain low, basal levels of lipid hydroperoxides on drying. The relationship between these observations and the antioxidant systems showed that the antioxidant enzymes CAT, ASC POD, AFR reductase, DHA reductase, guaiacol POD and SOD declined markedly during the unscheduled drying, whereas GSH and ASC declined only slightly. On rehydration, most of the enzymes, total glutathione, and total ascorbate pool increased, the only exception being the loss of ASC POD activity. ORA reductase, which was seen to decrease as a part of nonnal gennination, declined progressively also in H-D-R treatments. These results suggested that loss of viability was not attributable to a decline of the antioxidant systems but rather to the combined deleterious effects of increased lipid peroxidation, and a generalized and moderately compromised antioxidant system. These studies have indicated that the occurrence of lipid peroxidation can be seen as a nonnal part of seed development and gennination. The H-D-R studies, on the other hand, supported the concept that the balance between peroxidation reactions and the protective systems was critical to the development of desiccation tolerance. / Thesis (Ph.D.)-University of Natal, Durban, 2003.

In recent years research has shown that exposing seed to smoke or smoke solutions results in increased germination of some, but not all the species tested. Tests showed that exposing seeds of some commercial crops to smoke increased early plant growth. The stimulatory effects of smoke were shown to benefit the germination of species regardless of whether or not fire played a part of the species ecological cycle. In commercial forest nurseries any method of increasing the recovery rate of seed presents opportunities for realising savings of related production costs. Greater recovery rates of genetically improved seed present opportunities for capturing more related growth in field operations. Improving efficiencies of seed recovery at an early point in the production chain have multiplied effects further on down the chain. Two species of pine, namely Pinus taeda and Pinus elliottii that do not generally present high rates of germination were selected to test if applications of smoke solution could increase germination or emergence rates by more than 5%. As seed of both species are known to respond positively to existing seed pre-treatments the effects of smoke needed to be tested in combination, and apart from the pre-treatments. A secondary aim of the study was to examine the effect of smoke on early plant growth. Attempts to optimizes the concentration of the smoke solution were not undertaken as part of this study, as a rinsing treatment, included in the trials, is known to remove any inhibitory effects of a high concentration of the smoke solution. Tests to determine the variability of the seedlots was carried out for statistical purposes. The interaction between smoke application and pre-treatments were tested, firstly in Petri dishes under controlled environmental conditions, and then in nursery trays under standard commercial nursery conditions for both species. The inclusion of smoke in combination with the target moisture stratification (TMS and rinse pre treatment) had a significantly positive effect on P. taeda in a controlled environment. The same combination did not yield a positive results when tested under nursery conditions. Recommendations are made regarding future tests to see if the beneficial combination found in the controlled environment could be replicated under nursery conditions. Further motivation for conducting the tests exists in that the particular combination set gave significantly better early plant growth under nursery conditions. No other combinations tested yielded positive results. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2008.

Orientador: Sandra Maria Carmello Guerreiro / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-25T23:22:45Z (GMT). No. of bitstreams: 1 Milani_JulianaForesti_D.pdf: 4831941 bytes, checksum: 1e831abd54a3b76f7aa5f9a4b31ca239 (MD5) Previous issue date: 2014 / Resumo: A morfologia externa e interna dos frutos e sementes de Passifloraceae Juss. ex Roussel é pouco conhecida ou até desconhecida para muitas espécies. Estudos morfoanatômicos de frutos e sementes têm grande importância já que os mesmos exibem pequena plasticidade fenotípica. Neste contexto, este trabalho teve como objetivo elucidar a ontogenia do fruto e da semente de quatro espécies de Passiflora L. subgênero Decaloba (DC.) Rchb. superseção Decaloba (DC.) J.M. Macdougal & Feulliet seção Xerogona (Raf.) Killip que apresentam frutos do tipo cápsula, condição incomum na família. Para tanto, o material vegetal foi coletado e processado segundo técnicas convencionais em microscopia de luz e eletrônica, além da aplicação de técnica específica (TUNEL) para detecção de morte celular programada no estádio de deiscência do fruto. Foram estabelecidos estádios de desenvolvimento para os frutos e as sementes: Estádio I: Ovário e óvulo; Estádio II - fruto e semente em início de desenvolvimento: frequentes divisões celulares; Estádio III - fruto e semente jovens: alongamento celular; Estádio IV - fruto e semente maduros: diferenciação celular e deiscência do fruto. Nos estádios II a IV do fruto, o epicarpo e o endocarpo são uniestratificados e as maiores modificações decorrentes do desenvolvimento do fruto ocorrem no mesocarpo. Ao longo do desenvolvimento aumentam os espaços intercelulares a partir do endocarpo em direção ao epicarpo. Não há a formação de uma linha de deiscência. Os espaços intercelulares ocorrem devido à morte celular programada formando lacunas que acabam por romper o pericarpo. Os testes histoquímicos indicaram a presença de idioblastos contendo compostos fenólicos e proteínas no tecido fundamental do ovário e no mesocarpo. As sementes das espécies estudadas são bitegumentadas. A testa é constituída de duas camadas: exotesta e endotesta. O tégmen é composto por três camadas: exotégmen, mesotégmen e endotégmen. No início do desenvolvimento da semente observou-se o arilo de origem funicular, formado por células parenquimáticas e idioblastos contendo compostos fenólicos e amido. O desenvolvimento dos tegumentos da semente se dá pelo alongamento diferencial das células do exotégmen e da endostesta. Esse processo resulta na ruminação do endosperma. Na semente madura o tegumento externo formará a sarcotesta. Neste estádio, o exotégmen constitui a camada mecânica formada por macroesclereídes em paliçada, representando a esclerotesta. As informações encontradas no presente estudo revelam que as características morfoanatômicas de fruto e semente são bastante conservadas e unificadoras na seção. Diante do exposto, destaca-se a importância de novos estudos abrangendo mais espécies e abordando a evolução de caracteres e a inclusão de outros novos para facilitar a elucidação das relações infragenéricas de Passiflora que tem sido ampliada graças à cooperação de estudos morfoanatômicos e genéticos / Abstract: External and internal morphology of Passifloraceae Juss. ex Roussel fruits and seeds is little known or even unknown for many species. Morphological and anatomical studies of fruits and seeds have great importance since they exhibit little phenotypic plasticity. In this context, this study aimed to elucidate the fruit and seed ontogeny of four species of Passiflora L. Decaloba (DC.) Rchb. subgenus Decaloba (DC.) J.M. Macdougal & Feulliet supersection Xerogona (Raf.) Killip section that present capsule type of fruit, unusual condition in the family. For this, the plant material were collected and processed according to conventional techniques for light and electron microscopy; specific technique (TUNEL) were also applied to detect programmed cell death in the stage of fruit dehiscence. Four developmental stages were established: Stage I: Ovary and ovule; Stage II - fruit and seed in early development: frequent cell divisions; Stage III - young fruit and young seed: cell elongation; Stage IV - mature fruit and mature seed: cell differentiation and fruit dehiscence. In the stages II to IV of the fruit, epicarp and endocarp are unistratified and major changes from the development of the fruit occurs on mesocarp. During the development, the intercellular spaces increase from the endocarp towards the epicarp. There is no line of dehiscence. The intercellular spaces occur due to programmed cell death forming gaps that breaks the pericarp. Histochemical test indicated the presence of phenolic compounds and idioblasts containing proteins in fundamental tissue of ovary and mesocarp. The seeds of all species were bitegmic. The testa consists of two layers: exotesta and endotesta. The tegmen is composed of three layers: exotegmen, mesotegmen and endotegmen. At the beginning of seed development, aryl of funicular origin, formed by parenchyma cells that may containing phenolic compounds and starch, was observed. The development of the integument of the seed occurs by differential cell elongation of exotegmen and endostesta. This process results in a ruminate endosperm. In the mature seed, the outer integument forms the sarcotesta. At this stage, the exotegmen is the mechanical layer formed by macrosclereids in palisade, representing esclerotesta. Information found in this study reveals that morphoanatomical characteristics of fruit and seed are quite conserved and unified in the section. Given that, we highlight the importance of further studies including more species and addressing the evolution of characters and adding new ones to facilitate the elucidation of infrageneric relationships in Passiflora which has been expanded due to the cooperation of morpho-anatomical and genetic studies / Doutorado / Biologia Vegetal / Doutora em Biologia Vegetal

Bibliography: leaves 158-176. / xxii, 200 leaves : ill. (chiefly col.) ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Yield variation of faba bean in southern Australia is associated with seasonal and site variation in stress levels during podsetting and podfilling. This study examined the development of pods and seed growth in the field; quantified the effects of water stress and temperature on pod and seed development; quantified the importance of stem and podwall dry matter to seed growth and seed yield and examined the importance of genotype to these responses. / Thesis (Ph.D.)--University of Adelaide, Dept. of Plant Science, 1995

Bibliography: leaves 158-176. Yield variation of faba bean in southern Australia is associated with seasonal and site variation in stress levels during podsetting and podfilling. This study examined the development of pods and seed growth in the field; quantified the effects of water stress and temperature on pod and seed development; quantified the importance of stem and podwall dry matter to seed growth and seed yield and examined the importance of genotype to these responses.

Cowpea (Vigna unguiculata. L) is an important African crop. However, it is also an underutilized grain legume. Consequently, there is not enough research data on cowpea seed physiology. Whereas there is evidence of cowpea being a drought tolerant crop, there is no evidence to associate plant drought tolerance with seed quality in response to water stress. This study sought to understand the effect of production site and water stress on cowpea seed quality development with respect to germination capacity and vigour. Patterns of raffinose family of oligosaccharides (RFO) during seed development to mature dry stage were used to physiologically relate seed performance to water stress. The effect of water stress and exogenous ABA on the accumulation of stress LEA proteins (dehydrins) in relation to seed quality development and germination was investigated. RFOs are known for their roles in desiccation sensitivity but no studies have shown their significance in cowpeas. Seeds of six cowpea cultivars were produced at two distinct growth sites characterised by irrigated and dry land conditions. The seeds were assessed during six developmental stages, for water content, dry matter accumulation, and performance. Harvested seeds were then planted in a pot experiment under controlled conditions to examine the effect of water stress on seed quality development and data collected during three developmental stages. Harvested seeds from the pot experiment were subsequently analyzed for changes in RFO accumulation during development using gas chromatography. The seeds were also used to investigate the effect of water stress and ABA on the accumulation of stress LEA proteins (dehydrins) in relation to seed quality development in cowpea. In addition, this study evaluated the use of image analysis as a method that can be used to objectively determine seed coat colour variation in cowpea. Statistical variation in individual seed’s solute leakage for cowpea cultivars differing in seed coat colour and produced under different environmental conditions was explored and correlations were done between seed conductivity test with other aspects of seed performance during germination. Furthermore the results of the conductivity test were compared with accelerated aging test, in relation to seed performance. The study provided evidence that cowpea seed lots produced under different environmental, and possibly management conditions may not differ with respect to seed quality as determined by germination capacity and vigour. However, significant differences between sites with respect to seed maturation patterns determined by water content and dry matter accumulation were observed. Adverse maternal environmental effects on the subsequent performance of seeds in a drought tolerant crop may not necessarily lead to poor performance. Cultivar differences in response to simulated drought conditions at the whole plant and tissue level can be considerable and highly variable; however, these differences may not have adverse effects on the germination and vigour of the seeds. Drought avoidance mechanisms at the whole plant level in cowpea are quite efficient in allowing the species to adapt to simulated drought conditions. These mechanisms may allow the cowpea cultivars to maintain metabolism and restore conditions for their continued growth under water stress; and produce few seeds of high germination capacity and vigour. Stachyose was found to be the predominant member of the raffinose family of oligosaccharides in cowpea. It is suggested that stachyose accumulation could be used as an indicator of stress tolerance in cowpea. However, the relationship between RFO concentration and the acquisition of desiccation remained as a matter of speculation in the present study and is still generally inconclusive. There was no evidence to suggest the acquisition of maximum desiccation tolerance is associated with maximum seed vigour. It is suggested in cowpea, which is drought tolerant, that maximum vigour does not necessarily imply the acquisition of maximum desiccation tolerance; rather there is a minimum level of desiccation tolerance that is required for the development of optimal seed vigour. The use of an in vivo approach in the study of LEA function in cowpea enabled the accurate comparison of two different groups of LEA proteins in developing cowpea seeds under conditions of water stress and in relation to germination and vigour. Both group 1 LEA and group 2 LEA (dehydrin) were shown to increase in concentration in response to water stress. In addition group 1 LEA protein was observed to be relatively abundant in cowpea seeds. A maternal influence on LEA protein gene expression under conditions of water stress, which may induce dehydrin accumulation vii during the earlier stages of seed development, was implied by the observation that dehydrin-like proteins were induced after two weeks of development in cowpea plants subjected to stress during the vegetative phase. In addition, the exogenous application of ABA delayed radicle protrusion; this was associated with a delay in the disappearance of LEA proteins and is suggestive of a relationship between LEA protein accumulation and the acquisition of desiccation tolerance. The study has demonstrated that image analysis can objectively discriminate seed coat colour variation in cowpea. Dark coloured seeds in general performed better than light coloured seeds; however seed coat colour was not always associated with better performance. A newly developed Aging Stress Differential Index (ASDI) has been used in this study to demonstrate a link between seed coat colour and sensitivity to water stress. The ASDI correlated well with the observations relating stress tolerance to stachyose accumulation. The skewed distribution patterns in individual electrical conductivity and the presence of extreme values may have implications with respect to the suitability of using standard statistical analyses which compare mean values to evaluate such data. In addition variation in individual electrical conductivity may also be influenced by cultivar differences and the chemical composition of the seed coat. Therefore associations between seed coat colour and electrical conductivity as a measure of performance should be treated with caution. The AA test does reflect changes in seed vigour, however ranked electrical conductivity values after AA did not consistently reflect differences in seed performance between cultivars and sites, and they did not correlate well with other aspects of performance. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2007.

The aim of the study was to determine the effect of day/night temperatures (22/l6°C, 2712l oC and 33/27°C) and phosphorus levels (0, 0.12 and 1.2g per 20 kg soil) on seedling establishment and seed viability during three stages of seed development (15, 22 and 33 days after flowering) for seed of normal and quality protein maize cultivars. Soluble carbohydrate accumulation and mineral element content were determined using environmental scanning electron microscopy (ESEM) in relation to seed phytate levels and seed germination capacity at different stages of development. Leaf emergence rate and plant height during seed development were significantly (P < 0.05) influenced by temperature and phosphorus nutrition. Phosphorus in seed is stored primarily in the form of phytic acid, also known as phytate. Accumulation of phytate takes place during maturation phase of seed development. Phosphorus nutrition and temperature also caused a.significant (P < 0.05) increase in seed germination at all stages of seed development. Furthermore, phosphorus nutrition and temperature influenced occurrence of soluble carbohydrates in seeds. Myo-inositol, the sugar alcohol that forms the basic structure of phytate, was increased by P nutrition and increasing growth temperature. Whereas, QPM maize was generally found to perform poorly than normal maize, with respect to phytate content, seed germination and seedling establishment, both cultivars displayed the same responses to phosphorus nutrition and temperature. In both cultivars, globoids, the sites of phytate synthesis and storage, were found only in the embryonic axis. Subsequently, there were significantly low levels of mineral elements (P, Mg and K) found in the endosperm, compared with embryonic axis. This finding suggested that the embryonic axis plays a major role in seed performance, through its effects emanating from phyate metabolism. Myo-inositol plays a role in membrane biogenesis during stress conditions such as temperature by maintaining the integrity of the cell wall and minimizes the leaching of cations essential during germination. Myo-inositol, although it occurs in small concentrations, could be used to indicate seed quality in maize, because its accumulation was found to be associated with enhanced phyate levels and better seed germination in a wide range of temperatures. Low vigour seeds are associated with high electrolyte leakage during imbibition. Mineral elements form a significant portion of the imbibition leachate, which causes seeds to lose nutrients for early seedling growth. This study provided evidence that phosphorus nutrition can alleviate poor seed vigour of maize by improving phytate levels. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2006.

The International Seed Testing Association and the Association of Official Seed analysts define seed vigour as the ability of a seed lot to perform during the germination process and crop stand establishment under a wide range of environmental conditions. There are many ways to determine seed vigour, but few satisfy the requirements of being simple, inexpensive and reproducible, among others, to permit the seed industry to adopt seed vigour as an indicator of seed quality when they sell seeds. Hence, the standard germination test, which is performed under uniform and favourable conditions, is generally used to indicate seed quality when seeds are marketed. The objective of this study was to determine the performance of tomato and pepper seeds in response to pre-germination hydration and dehydration relative humidities (12%, 49% and 75% RH). Before hydration, seeds were hydrated at three temperatures (10°C, 20°C and 30°C). Hydration was performed by imbibing pre-weighed pepper ('Santarini' and 'California Wonder'), Chilli and tomato ('Heinz', 'Marondera' and 'Roma') seeds for 2 h in 10 ml of distilled H(2)0 per 100 seeds at 10°C, 20°C and 30°C. Dehydration was performed by change in seed mass determined during a 72-hour dehydration at 12%, 49% and 75% RH. Seed performance in response to imbibition and dehydration was determined by leakage of electrolytes from seeds during imbibition, laboratory germination capacity and seedling emergence under simulated shadehouse nursery conditions. A pot experiment was conducted to determine the effects of seed treatments on yield. Seed mass increased by about 50% during the 2-hour of hydration. Dehydration was hastened by decreasing the RH, and 12% RH significantly (P < 0.05) reduced the post-imbibition seed moisture content compared with 49% and 75% RH. The latter two relative humidities reduced the seed moisture content to about 10% and 15%, respectively, for all cultivars, irrespective of imbibition temperature. Low imbibition temperature (10°C) significantly (P < 0.01) increased electrolyte leakage, compared with high imbibition temperatures (20°C and 30°C), which were not significantly different from each other. At all hydration temperatures, low RH (12%) caused a significant (P < 0.01) decrease in seed germination whereas 49% RH and 75% RH apparently had a priming effect on seeds. There was no significant difference between imbibition temperatures, with respect to seed germination, but 100G caused a significant decrease in germination index, a measure of seed vigour. Seedling emergence was significantly (P < 0.01) reduced by both low imbibition temperature (10oG) and low dehydration relative humidity (12% RH). The negative effects of low imbibition temperature and rapid dehydration at 12% RH were also observed as stunted seedling growth. Principal component analysis and linear regression were used to determine a statistical model to predict seedling emergence from germination percentage. The model predicted emergence consistently, but it overestimated it by about 2% to 3%. It is concluded that low imbibition temperature and rapid dehydration can be used to simulate stress to determine seed performance in pepper and tomato. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2005.

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.