Submitted by Ricardo Cedraz Duque Moliterno (ricardo.moliterno@uefs.br) on 2016-08-24T20:52:09Z No. of bitstreams: 1 Disserta??o Final_Fl?via_PPGRGV (2).pdf: 1857571 bytes, checksum: 967f2b7d728622a11a98a803f13ff0de (MD5) / Made available in DSpace on 2016-08-24T20:52:09Z (GMT). No. of bitstreams: 1 Disserta??o Final_Fl?via_PPGRGV (2).pdf: 1857571 bytes, checksum: 967f2b7d728622a11a98a803f13ff0de (MD5) Previous issue date: 2015-03-20 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Hyptis ramosa Pohl ex Benth (Lamiaceae) is native and endemic to the semi-arid northeast, with its unknown phytochemical constitution so far. Considering the pharmacological importance of the species of this family, the development of forms of propagation and in vitro culture can contribute to the inclusion of these species in sustainable production systems and the conservation of the same. Thus, the objective of this work was to study the in vitro propagation of the species H. ramosa, through direct organogenesis and callus formation, and the biochemical characterization of the obtained calluses, thus allowing the establishment of strategies for their conservation and sustainable use. To this, H. ramosa seeds were disinfected and established in medium MS / 2 culture. In the multiplication phase was tested the influence of cytokinins BAP, CIN and TDZ on different explants. The obtained shoots were individualized and transferred to MS / 2 medium containing different concentrations of auxin IBA and activated carbon for rooting them. Regenerated and rooted in vitro microplants were subjected to pre-acclimatization in different cultivation container closure and then were transferred to ex vitro conditions in commercial substrate Plantmax? being quantified plant survival rate at 30 days after transfer. For callus induction, we used explants and different concentrations of 2,4-D and BAP, determining the growth curve from the fresh weight of callus until the 28th day of cultivation, at intervals of seven days. Concurrent with obtaining the growth curve it is quantified in calluses obtained the total soluble sugar content, reducing sugar and crude protein. The in vitro propagation of H. ramosa is possible using the nodal segment as a source of explants in MS medium supplemented with BAP. In vitro rooting occurs, even in free auxin. The species showed survival of 100%, regardless of the day of pre-acclimatization phase. For callus induction the best explant is the nodal segment, and the combination of 2.4-D and BAP favor the formation of the same. The callus growth curve showed quadratic behavior with two different phases and biochemical analysis showed the maximum level of total soluble sugars, reducing sugars and crude protein at 14 ?, 21 ? and 14 ? days, respectively. / Hyptis ramosa Pohl ex Benth (Lamiaceae) ? uma esp?cie nativa e end?mica do semi?rido nordestino, sendo sua constitui??o fitoqu?mica desconhecida at? o momento. Considerando a import?ncia farmacol?gica das esp?cies dessa fam?lia, o desenvolvimento de formas de propaga??o e cultivo in vitro poder? contribuir para a inser??o dessas esp?cies em sistemas de produ??o sustent?veis e a conserva??o das mesmas. Diante disso, o objetivo deste trabalho foi estudar a propaga??o in vitro da esp?cie H. ramosa, atrav?s de organog?nese direta e calog?nese, bem como a caracteriza??o bioqu?mica dos calos obtidos, permitindo assim o estabelecimento de estrat?gias para a sua conserva??o e explora??o sustent?vel. Para isso, sementes de H. ramosa foram desinfestadas e estabelecidas em meio de cultura MS/2. Na fase de multiplica??o foi testada a influ?ncia das citocininas BAP, CIN e TDZ sobre diferentes explantes. As brota??es obtidas foram individualizadas e transferidas para meio MS/2 contendo diferentes concentra??es da auxina AIB e de carv?o ativo para o enraizamento das mesmas. As microplantas regeneradas e enraizadas in vitro foram submetidas ? pr?-aclimatiza??o em diferentes tipos de fechamento do recipiente de cultivo e, posteriormente, foram transferidas para a condi??o ex vitro em substrato comercial Plantmax?, sendo quantificada a taxa de sobreviv?ncia das plantas aos 30 dias ap?s a transfer?ncia. Para a indu??o de calos utilizou-se diferentes explantes e concentra??es de 2,4-D e BAP, determinando-se a curva de crescimento a partir da mat?ria fresca dos calos at? o 28o dia de cultivo, em intervalos de sete dias. Concomitante com a obten??o da curva de crescimento quantificou-se nos calos obtidos o teor de a??cares sol?veis totais, a??cares redutores e prote?na bruta. A propaga??o in vitro de H. ramosa ? poss?vel utilizando-se o segmento nodal como fonte de explante, em meio de cultura MS suplementado com BAP. O enraizamento in vitro ocorre, mesmo em meio isento de auxina. A esp?cie apresentou sobreviv?ncia de 100%, independentemente da realiza??o da fase de pr?-aclimatiza??o. Para indu??o de calos o melhor explante ? o segmento nodal, sendo a combina??o de 2.4-D e BAP favor?vel a forma??o dos mesmos. A curva de crescimento de calos mostrou comportamento quadr?tico com duas fases distintas e a an?lise bioqu?mica evidenciou o teor m?ximo de a??cares sol?veis totais, a??cares redutores e prote?na bruta aos 14?, 21? e 14? dias, respectivamente.

Submitted by Ricardo Cedraz Duque Moliterno (ricardo.moliterno@uefs.br) on 2016-09-27T00:25:31Z No. of bitstreams: 1 VERS?O FINAL DISSERTA??O-Luma Passos.pdf: 2866580 bytes, checksum: 631bf5534fde43f51e60719229aabc72 (MD5) / Made available in DSpace on 2016-09-27T00:25:31Z (GMT). No. of bitstreams: 1 VERS?O FINAL DISSERTA??O-Luma Passos.pdf: 2866580 bytes, checksum: 631bf5534fde43f51e60719229aabc72 (MD5) Previous issue date: 2015-03-31 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Funda??o de Amparo ? Pesquisa do Estado da Bahia - FAPEB / The genus Lippia (Verbenaceae) is characterized by having species with medicinal and aromatic properties, its economic potential aims to the production of essential oils with important biological and pharmacological activities. Even with all known diversity and potential, few species are the target of agronomic studies. Considering the propagation as the first step in the domestication of a species and the influence of external factors on essential oil production, this study aimed to evaluate the effect of cutting type and auxin concentrations in the vegetative propagation of the species L. insignis, L. lasiocalycina and L. thymoides, as well as different types of fertilization and harvest season in production, and content and chemical composition of the essential oil. In propagation tests two types of cuttings were used (apical and middle) and five IBA concentrations (0, 250, 500, 750 and 1000 mg L-1). In cultivation three forms of fertilization were tested (control, manure and manure + NPK 10:10:10) and two harvest seasons (210 and 360 days after planting). The essential oils are extracted from dry leaves by the hydrodistillation method by Clevenger apparatus, and chemical composition determined by gas chromatography with flame ionization detector (GC / FID) coupled to mass spectrometer (GC / MS). It was concluded that apical cuttings are the most suitable for the propagation of L. insignis and both apical and middle cuttings for L. thymoides and L. lasiocalycina; treatment with IBA is not necessary in the propagation by cuttings of these species; chemical fertilizers (NPK) associated with cattle manure was the best treatment in growth and biomass production of the species L. lasiocalycina and L. thymoides, while for L. insignis to the control treatment was the best; pruning, followed by the second growing season (regrowth) is recommended in the cultivation of the studied species, aiming at the production of essential oil; ?-myrcene, limonene and E-ocimenone are the major components of L. insignis; ?-myrcene and E-ocimenone the major components of L. lasiocalycina and E-caryophyllene and caryophyllene oxide the major components of L. thymoides; fertilization and harvest season promote quantitative variations in the chemical composition of essential oils; the species L. insignis and L. thymoides are the most promising for essential oil production in the conditions of Feira de Santana, Bahia, Brazil. / O g?nero Lippia (Verbenaceae) ? caracterizado por apresentar esp?cies com propriedades medicinais e arom?ticas, sendo seu potencial econ?mico voltado para a produ??o de ?leos essenciais com importantes atividades biol?gicas e farmacol?gicas. Mesmo com toda diversidade e potencial conhecidos, poucas esp?cies est?o sendo alvo de estudos agron?micos. Considerando a propaga??o como o primeiro passo na domestica??o de uma esp?cie e a influ?ncia dos fatores externos na produ??o de ?leo essencial, o presente trabalho teve como objetivo avaliar o efeito do tipo de estaca e concentra??es de auxina na propaga??o vegetativa das esp?cies L. insignis, L. lasiocalycina e L. thymoides, bem como de diferentes tipos de aduba??o e ?pocas de colheita na produ??o, teor e composi??o qu?mica do ?leo essencial. Nos testes de propaga??o foram utilizados dois tipos de estacas (apical e mediana) e cinco concentra??es de AIB (0, 250, 500, 750 e 1000 mg L-1). No cultivo foram testadas tr?s formas de aduba??o (testemunha, esterco bovino e esterco + NPK 10:10:10) e duas ?pocas de colheita (210 e 360 dias ap?s o plantio). Os ?leos essenciais foram extra?dos de folhas secas pelo m?todo de hidrodestila??o por aparelho de Clevenger e a composi??o qu?mica determinada por cromatografia gasosa com detector de ioniza??o em chama (CG/DIC) acoplado a espectr?metro de massas (CG/EM). Foi poss?vel concluir que estacas apicais s?o as mais indicadas na propaga??o de L. insignis e tanto as estacas apicais quanto medianas para L. thymoides e L. lasiocalycina; o tratamento com AIB n?o ? necess?rio na propaga??o por estaquia dessas esp?cies; a aduba??o mineral (NPK) associado a esterco bovino foi o melhor tratamento no crescimento e produ??o de biomassa das esp?cies L. lasiocalycina e L. thymoides, j? para L. insignis o tratamento testemunha foi o melhor; a poda, seguida da segunda ?poca de cultivo (rebrota) ? recomend?vel no cultivo das esp?cies estudadas, visando ? produ??o de ?leo essencial; ?-mirceno, limoneno e E-ocimenona s?o os componentes majorit?rios de L. insignis; ?-mirceno e E-ocimenona os componentes majorit?rios de L. lasiocalycina e E-cariofileno e ?xido de cariofileno os componentes majorit?rios de L. thymoides; a aduba??o e a ?poca de colheita promovem varia??es quantitativas na composi??o qu?mica dos ?leos essenciais; as esp?cies L. insignis e L. thymoides s?o as mais promissoras na produ??o de ?leo essencial nas condi??es de Feira de Santana, Bahia, Brasil.

La résistance aux antimicrobiens est un problème mondial majeur de plus en plus préoccupant. Le développement de nouveaux antimicrobiens classiques remplaçant les agents rendus inefficaces est certes l’une des stratégies les plus prometteuses visant à diminuer son ampleur. Mais cette solution demeure malheureusement momentanée. En fait tout agent antimicrobien sera tôt ou tard vaincu par la propagation des souches résistantes favorisée par le phénomène de pression de sélection exercée par l’agent lui-même. Il est donc nécessaire d’envisager toutes les alternatives possibles aux approches classiques, cela afin de diversifier l’arsenal thérapeutique antimicrobien.Les huiles essentielles utilisées par les plantes dans leur défense contre les pathogènes sont naturellement composées d’un mélange de constituants les rendant capables d’agir sur plusieurs cibles de l’organisme. De ce fait, même les pathogènes les plus résistants ne vont pouvoir échapper à leurs actions et le développement des résistances sera par conséquent limité. Dans le cadre de la présente thèse des plantes aromatiques libanaises sélectionnées en se basant surtout sur des indications ethno pharmacologiques ont été récoltées de différentes régions libanaises. Elles ont été par la suite hydrodistillées pour produire une vingtaine d’huiles essentielles qui ont été analysées par GC/MS et éventuellement par RMN, et évaluées contre une gamme de pathogènes responsables d’infections cutanées chez l’homme. L’analyse de la composition chimique des HEs a été marquée par l’isolement et la caractérisation pour la première fois du santolinoïdol, un sesquiterpène du type bisabolène qui a été retrouvé dans l’huile essentielle d’Achillea santolinoides subsp. wilhelmsii. Presque la moitié des HEs ont été actives sur au moins un des pathogènes. L’origine de l’activité des huiles les plus actives, l’effet de leurs associations avec des antimicrobiens commerciaux ainsi que leurs cytotoxicités ont été également investigués. / Antimicrobial resistance is a major global problem and a growing concern. The development of new conventional antimicrobial agents replacing inefficient ones is certainly one of the most promising strategies aiming to reduce its extent. But this solution remains unfortunately momentary. In fact, any antimicrobial agent will eventually be defeated by the spread of resistant strains favored by the selection pressure phenomenon exerted by the agent itself. It is therefore necessary to consider all possible alternatives to conventional approaches in order to diversify the antimicrobial arsenal.Essential oils used by plants in their defense against pathogens are naturally composed of a mixture of components making them able to act on several targets of the organism. Thus, even the most resistant pathogens will not be able to escape their actions and the development of resistance will be therefore limited.In this thesis, Lebanese aromatic plants selected based mainly on ethnopharmacological indications were collected from various Lebanese regions. They were subsequently hydrodistillated producing twenty essential oils analyzed by GC / MS and eventually by NMR, and assessed against a range of pathogens that cause skin infections in humans. The analysis of the chemical composition of the EOs was marked by the isolation and characterization for the first time of santolinoïdol, a bisabolene type sesquiterpene that was found in the essential oil of Achillea santolinoides subsp. wilhelmsii. Almost half of the EOs have been active on at least one pathogen. The origin of the activity of the most active oils, the effect of their associations with commercial antimicrobials and their cytotoxicities were also investigated.

Text in English / The introduction of the New Growth Path (NGP) and the New Industrial Policy Framework (NIPF) have created an opportunity for the industrialisation of indigenous medicinal and aromatic plants (MAPs Amarula (Sclerocarya birrea), Buchu (Agathosma betulina), and Pelargonium (Pelargonium sidoides) for job creation and economic development in South Africa. The aim is to create a locally sustainable industry which creates jobs as well as improvement of the balance of payments. The National Growth Plan highlights some of the opportunities for industrialization as well as creation of decent jobs. There is increasing pressure on the harvesting of wild indigenous medicinal and aromatic plants (MAPS). The establishment of a local pharmaceutical and nutraceutical industry for processing MAPS remains a challenge in South Africa and many low income countries. The objective of this study was to assess the challenges and their impact looking at particular species, Amarula, Buchu and Pelargonium by using structured focus group interviews with users in communities and businesses in KwaZulu Natal Province, Eastern Cape and the Western Cape. More data/information was also collected from key stakeholder companies including desk-top reviews. Data/information was analysed using the SPSS computer programme and the Atlas software to summarize data and the results. Graphs and tables were used where necessary. The qualitative research reported on some qualitative assessments and challenges facing the establishment of medicinal and aromatic plants in South Africa. Comparative findings from BRICS were also reported. The results of the study indicated that there is continuous exploitation of several medicinal plant species, for plant trade, from the wild and substantial loss of their habitat. The research identifies a number of challenges such as: poor standards for raw materials, lack of research and development linking industry driven revenues, demand exceeding supply, regulatory risk, community risk, lack of understanding of judicious agronomic practices, ethical challenges, liquidity risk as well as political instability fail to attract foreign direct investments. The research concludes that for the establishment of a viable local pharmaceutical industry, iii beverage industry and nutraceutical industry with sustainable harvesting, there are serious hegemonic challenges which will require multi-stakeholder analysis. However, for the South African (SA) industry to be competitive, detailed feasibility studies need to be done comparing SA with other BRICS countries. There is also a need for the Research and Development to be strengthened. It must also be linked into industrial development in the industry. Furthermore there is a need to initiate the incentivizing of the pharmaceutical manufacturing to lower the cost of setting up the businesses in the industry and lessen the need to harvest from South African forests. Furthermore there is a need for programmes of production so that key species are given opportunity for cultivation and the attention they deserve to preserve these depleting resources. / Agriculture and  Animal Health / M.Sc. (Agriculture)

Ce travail a été effectué dans le cadre d’une convention de cotutelle entre les Universités de Corse et d’Antananarivo. Il a contribué à la caractérisation de cinq Plantes de Madagascar à travers la composition chimique de leurs huiles essentielles (HE). Quatre plantes aromatiques et médicinales (Cymbopogon giganteus var. madagascariensis, Tana bojeriana, Croton kimosorum et Croton sp.) poussent à l’état spontané dans la région sub-aride du sud-ouest de Madagascar (Toliara) alors que l’espèce Vepris madagascarica est présente dans la forêt dense humide du nord-est (Vohémar). Ainsi, nous avons étudié par combinaison CPG (IR), CPG-SM et RMN 13C la composition chimique d’huiles essentielles obtenues par hydrodistillation (appareil type Clevenger) et par entraînement à la vapeur d’eau (Alambic).Les huiles essentielles de Cymbopogon giganteus Chiov. var. madagascariensis (A. Camus) ou Ahibero possèdent une composition chimique comparable avec celles des espèces Cymbopogon giganteus Chiov. africaines décrites dans la littérature. Elles sont dominées par les quatre menthadiénols (cis- et trans-p-mentha-1(7),8-dièn-2-ol, cis- et trans-p-mentha-2,8-dièn-1-ol) et le limonène. Nous avons aussi identifié quatre hydropéroxydes possédant le squelette p-menthane. Une monographie de l’huile essentielle d’Ahibero a été proposée à l’intention des utilisateurs de cette huile essentielle.La composition chimique de l’HE de Tana bojeriana Baker est caractérisée par la présence majoritaire de l’-phellandrène (30,4%), de la carvone (22,3%), du limonène (13,3%) et du dill éther (7,7%). Cette composition chimique apparait très proche de celle d’Anethum graveolens (Apiaceae). La présence du trans-p-menth-2-èn-1,6-diol et du p-mentha-1(7),2-dièn-6-ol lui confère une certaine originalité Elle pourrait donc constituer une alternative aux huiles essentielles qui sont bien implantées sur le marché des Plantes Aromatiques et Médicinales (PAM).La composition chimique de l’HE de feuilles de Vepris madagascarica (Baill.) H. Perrier est caractérisée par une très forte teneur en (E)-anéthole (78,2%). L’étude comparative de la composition chimique des huiles essentielles de feuilles (fraîches et séchées) et d’écorces de tronc confirme la prédominance du (E)-anéthole (69,5 à 84,6%). Compte tenu de la répartition restreinte de cette plante aromatique et en vue d’une gestion durable de cette espèce, son exploitation devrait être limitée aux huiles essentielles des feuilles qui pourraient ainsi constituer une source de (E)-anéthole, composé très utilisé en agro-alimentaire.Deux espèces de Croton endémiques de Madagascar ont été étudiées, Croton kimosorum Leandri (Zanapoly ou Napoly) et Croton sp. (Andriambolamena ou Riamena). Les huiles essentielles de ces deux plantes aromatiques et médicinales sont caractérisées par la présence du linalol (Zanapoly) et du limonène(Andriambolamena). La première HE est caractérisée par une majorité de composés oxygénés tandis que la seconde est dominée par les composés oléfiniques monoterpéniques. Dans les deux cas, de nombreux sesquiterpènes oxygénés ont été identifiés. Parmi eux, l’épi--bisabolol, jamais décrit dans une HE du genre Croton, est trouvé pour la première fois dans les deux espèces. / This study was conducted under an agreement of co-supervision between the Universities of Antananarivo (Madagascar) and Corsica (France). It contributed to the characterization of five malagasy plants through the chemical composition of their essential oils (EO). Four aromatic and medicinal plants: Cymbopogon giganteus var. madagascariensis, Tana bojeriana, Croton kimosorum and Croton sp. grow wild in the south-western region of Madagascar (Toliara) whereas the species Vepris madagascarica is widespread in the rainforest of the north-eastern region (Vohémar). The identification of components of EOs produced by hydrodistillation (Clevenger type apparatus) and by steam distillation (Alembic) has been obtained by combination of GC (RI), GC-MS and 13C NMR.Cymbopogon giganteus Chiov. var. madagascariensis (A. Camus) EO or Ahibero EO of Madagascar has similar chemical composition with Cymbopogon giganteus Chiov. african species described in literature. Its chemical composition is dominated by four p-menthadienols (cis- and trans-p-mentha-1(7),8-dien-2-ol, cis- and trans-p-mentha-2,8-dien-1-ol) and limonene. Moreover, four peroxides derivatives bearing the p-menthane skeleton were also identified. A monography has been proposed to Ahibero EO users.Tana bojeriana Baker EO is characterized by the predominance of -phellandrene (30.4%), carvone (22.3%), limonene (13.3%) and dill ether (7.7%). The chemical composition of this EO was very close to that of Anetum graveolens (Apiaceae family), constituents. The presence of trans-p-menth-2-en-1,6-diol and p-mentha-1(7) ,2-dien-6-ol, gives some originality to Tana bojeriana EO. It could be therefore an alternative for essential oils that are already well established on the Medicinal and Aromatic Plants market.The chemical composition of Vepris madagascarica (Baill.) H. Perrier leaf EO is characterized by a high content of (E)-anethole (78.2%). The comparative study of the chemical composition of leaves (fresh and dried) and bark trunk EOs confirmed the predominance of (E)-anethole (69.5 to 84.6%). Taking account of the restricted distribution of this aromatic plant and the need for sustainable management of this species, its potential use would be limited to leaf oil and could be a source for this compound widely used in food industry.Two species of Croton endemic from Madagascar were studied, Croton kimosorum Leandri (Zanapoly or Napoly) and Croton sp. (Andriambolamena or Riamena). Both aromatic and medicinal plants EOs are characterized by the presence of linalool (Zanapoly) and limonene (Andriambolamena). Croton kimosorum is characterized by a majority of oxygenated compounds, while the second is dominated by hydrocarbon monoterpenes. In both cases, several oxygenated sesquiterpenes are identified. Among these, the epi--bisabolol, never reported in of Croton EOs, was found for the first time in both species.

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.

Text in English / The introduction of the New Growth Path (NGP) and the New Industrial Policy Framework (NIPF) have created an opportunity for the industrialisation of indigenous medicinal and aromatic plants (MAPs Amarula (Sclerocarya birrea), Buchu (Agathosma betulina), and Pelargonium (Pelargonium sidoides) for job creation and economic development in South Africa. The aim is to create a locally sustainable industry which creates jobs as well as improvement of the balance of payments. The National Growth Plan highlights some of the opportunities for industrialization as well as creation of decent jobs. There is increasing pressure on the harvesting of wild indigenous medicinal and aromatic plants (MAPS). The establishment of a local pharmaceutical and nutraceutical industry for processing MAPS remains a challenge in South Africa and many low income countries. The objective of this study was to assess the challenges and their impact looking at particular species, Amarula, Buchu and Pelargonium by using structured focus group interviews with users in communities and businesses in KwaZulu Natal Province, Eastern Cape and the Western Cape. More data/information was also collected from key stakeholder companies including desk-top reviews. Data/information was analysed using the SPSS computer programme and the Atlas software to summarize data and the results. Graphs and tables were used where necessary. The qualitative research reported on some qualitative assessments and challenges facing the establishment of medicinal and aromatic plants in South Africa. Comparative findings from BRICS were also reported. The results of the study indicated that there is continuous exploitation of several medicinal plant species, for plant trade, from the wild and substantial loss of their habitat. The research identifies a number of challenges such as: poor standards for raw materials, lack of research and development linking industry driven revenues, demand exceeding supply, regulatory risk, community risk, lack of understanding of judicious agronomic practices, ethical challenges, liquidity risk as well as political instability fail to attract foreign direct investments. The research concludes that for the establishment of a viable local pharmaceutical industry, iii beverage industry and nutraceutical industry with sustainable harvesting, there are serious hegemonic challenges which will require multi-stakeholder analysis. However, for the South African (SA) industry to be competitive, detailed feasibility studies need to be done comparing SA with other BRICS countries. There is also a need for the Research and Development to be strengthened. It must also be linked into industrial development in the industry. Furthermore there is a need to initiate the incentivizing of the pharmaceutical manufacturing to lower the cost of setting up the businesses in the industry and lessen the need to harvest from South African forests. Furthermore there is a need for programmes of production so that key species are given opportunity for cultivation and the attention they deserve to preserve these depleting resources. / Agriculture and  Animal Health / M. Sc. (Agriculture)

Les plantes aromatiques font parties des richesses naturelles du patrimoine de la région Midi-Pyrénées mais apparaissent cependant de moins en moins valorisées. Dans le cadre de la recherche de nouveaux aromatisants naturels de type WONF, nous avons choisi d'adopter la voie de valorisation des plantes aromatiques médiévales de cette région car elles ont toujours fait l'objet de cueillette à l'état sauvage sans jamais être cultivées. Dans une première étape, une étude bibliographique a été réalisée, à partir d'anciens ouvrages botaniques et de bases de données scientifiques, afin d'identifier les plantes régionales traditionnellement utilisées par nos ancêtres pour leurs activités aromatisantes, culinaires ou médicinales. Nous avons sélectionné cinq plantes aromatiques : Achillea millefolium, Calamintha grandiflora, Tanacetum balsamita, Myrrhis odorata et Monarda didyma. Dans une deuxième étape, des études physico-chimiques, sensorielles et biologiques des fractions volatiles et non volatiles, obtenues après extractions des plantes médiévales sélectionnées, ont été réalisées pour juger de leurs activités aromatiques, antioxydantes et antibactériennes ainsi que de leur intérêt dans le cadre de nouvelles réglementations européennes. Les résultats de ces travaux montrent des activités odorantes, antioxydantes et antibactériennes significatives pour l'ensemble des plantes sélectionnées. Dans une troisième étape, du fait de la rareté du végétal étudié, nous nous sommes intéressés à deux voies analytiques d'évaluation rapide de la composition de l'huile essentielle native de ces plantes aromatiques ; premièrement au moyen d'un échantillonnage classique couplé à un détecteur à forte sensibilité comme la chromatographie bidimensionnelle et deuxièmement par un échantillonnage spécifique couplé à un détecteur classique pour lequel nous avons développé un appareillage novateur dénommé "Crushing Finger Device" (CFD). Ce dernier permet de mimer instrumentalement l'écrasement de la feuille entre les doigts et de piéger les composés volatils odorants émis en vue de leur analyse chromatographique ultérieure. Enfin, pour une application pré-industrielle, afin d'élargir le champ d'application de ces plantes aromatiques, des études d'infusion des plantes et d'aromatisation des huiles végétales par les huiles essentielles ont été mises en place. Durant le processus d'aromatisation de l'huile végétale, les volatils émis par la plante ou par l'huile essentielle donnent un goût et une saveur spécifique tout en délivrant une activité biologique notable. / Midi-Pyrenees region has a rich and unvalorized wild vegetal heritage, especially for aromatic plants. In the framework of the search for new natural flavorings e.g. WONF, we chose to adopt the way of valorization of the medieval aromatic plants of this region, because they have been always picked in a wild state without being ever grown. In a first step, a global bibliographical survey study based on historical books, library archived documents and computerized data banks, was made in order to identify regional plants which were used by our ancestors for their odorous, culinary or medicinal activities. We selected five aromatic plants, e.g. Achillea millefolium, Calamintha grandiflora, Tanacetum balsamita, Myrrhis odorata and Monarda didyma. In a second step, physico-chemical, sensory and biological studies of volatiles and non volatiles fractions, obtained by extractions of the selected medieval plants were carried out, to judge their aromatic, antioxidant and antibacterial activities and their interest within the framework of new european regulations. Theses results show a significant odorous, antioxidant and antimicrobial activities for all selected plants. In a third step, such plants were rare or their harvesting problematic, inducing a limited sampling. We decided to investigate two strategic approaches to perform a rapid evaluation of native essential oil, based either on a classic sampling with a specific/ultra sensitive detection such as Comprehensive GC, nor the use of a specific/high effective sampling system with a classic detection. This last methodology was based on a self designed apparatus “Crushing Finger Device” (CFD), allowing to copy the crushing of leaves between fingers and to trap the volatiles emitted to their later chromatographic analysis. Finally, in the aim to expand the use of aromatic plants, also to enlarge the assortment of oil products, a pre-industrial application were carried out. Infusion of plants and flavoring of vegetable oil by essential oil were studied. During processing of aromatized vegetable oil, volatiles from added aromatic plants or essential oil migrate to oil to give a specific aroma and a significant biological activity. Plantes aromatiques oubliées, Huile essentielle, Antioxydant, Antibactérien, Simulateur de froissement, Composés d’arômes, Achillea millefolium, Calamintha grandiflora, Tanacetum balsamita, Myrrhis odorata, Monarda didyma.

Les régions de Midi-Pyrénées (France) et de Chongqing (Chine) sont riches en plantes aromatiques et médicinales dites oubliées (ou médiévales). Afin de valoriser pleinement les différentes bio-molécules extractibles de ces plantes, le concept de MAP-raffinerie a été créé et appliqué à une sélection de plantes issues de ces deux régions. Plusieurs technologies d’extraction utilisant l’eau comme solvant vert (hydrodistillation, distillation à la vapeur et extraction par eau sub-critique) ont ainsi été employées et leur impact tant sur la composition des huiles essentielles que sur la récupération des molécules anti-oxydantes a été évalué. Dans un premier temps, une liste de plantes aromatiques et médicinales oubliées, voire sous-utilisées dans les deux régions a été établie selon des règles de sélection prédéfinies. Six plantes modèles de la région de Midi-Pyrénées (Tussilago farfara L., Calendula arvensis L., Robinia pseudoacacia L., Geranium robertianum L., Cytisus scoparius L. et Spartium junceum L.) et trois plantes de la région de Chongqing (Tussilago farfara L., Citrus aurantium L. et Saussurea costus) ont finalement été retenues. Puis, le concept de MAP-raffinerie a été appliqué à ces plantes afin d’étudier leur possible valorisation globale. L’étude des compositions chimiques des extraits volatils des racines de Tussilago farfara L. et de Calendula arvensis L., ainsi que des boutons de fleurs de Spartium junceum L. a été réalisée par GC et GC-MS pour la première fois. Les principaux composés chimiques dans l’extrait volatil de racines de Tussilago farfara L. étaient des hydrocarbures sesquiterpéniques et des composés aliphatiques tandis que les principaux composés chimiques dans l’extrait volatil de racines de Calendula arvensis L. étaient des sesquiterpènes oxygénés, des monoterpènes oxygénés et des diterpènes oxygénés. L’extrait volatil de boutons de fleurs de Spartium junceum L. était principalement composé de composés aliphatiques. Par ailleurs, les résultats de l’évaluation des capacités anti-oxydantes des extraits (par les tests DPPH, ABTS, FRAP, ORAC et Folin-Ciocalteu) ont montrés que plusieurs plantes comme Cytisus scoparius L., Tussilago farfara L., Citrus aurantium L. ou Robinia pseudoacacia L. pourraient être des sources potentielles d’anti-oxydants naturels. D’un point de vue technologique, les comparaisons de l’utilisation de l’hydrodistillation (HD), de la distillation à la vapeur (SD) et de l’extraction par eau sub-critique (SWE) ont montrées que si la HD et la SD ont des effets limités sur la composition des huiles essentielles, la HD semble être une méthode plus efficace pour la récupération des composés anti-oxydants à partir des résidus de distillation que la SD tandis que la SWE s’avère être une technologie prometteuse pour l’extraction directe de ces molécules à partir des plantes. Si la composition minérale de l’eau lors de l’hydrodistillation n’a que des effets très limités sur les rendements d’extraction, les teneurs en ions calcium et bicarbonate des eaux ont par contre des effets décroissants significatifs sur la capacité anti-oxydante et sur la teneur phénolique totale des extraits aqueux et méthanoliques. Au vue de ces résultats, un concept amélioré de MAP-raffinerie a été développé en intégrant une extraction à l’eau sub-critique pour l’extraction des composés anti-oxydants des résidus d’extraction primaire. Selon ce nouveau concept, cinq extraits peuvent être obtenus à partir des matières végétales: un extrait volatil, un extrait aqueux, un extrait méthanolique, un extrait à l’eau sub-critique et in fine un résidu solide. Les premiers résultats ont montrés que la "MAP-raffinerie améliorée" augmente de manière significative la récupération des antioxydants par rapport à la MAP-raffinerie originale et permet d’envisager une valorisation plus facile du résidu solide en agro-matériaux du fait de sa faible teneur en eau résiduelle. / In both Midi-Pyrénées region (France) and Chongqing region (China), there are rich and underutilized medicinal and aromatic plants (MAP). Aiming at fully exploiting different molecules in these plants, the concept of MAP-refinery was developed and applied to several underutilized medicinal and aromatic plants in these two regions. Several water-based green extraction technologies of natural products (e.g. hydrodistillation, steam distillation and subcritical water extraction) were also investigated to look at their effects on essential oil composition and antioxidants recovery from selected plants. Firstly, lists of forgotten or underutilized medicinal and aromatic plants in both regions were established according to the rules of selection. From the lists, six plants in the Midi-Pyrénées region (Tussilago farfara L., Calendula arvensis L., Robinia pseudoacacia L., Geranium robertianum L., Cytisus scoparius L. and Spartium junceum L.) and three plants in the Chongqing region (Tussilago farfara L., Citrum aurantium L. and Saussurea costus) were finally selected for investigations. Then the MAP-refinery was applied to the selected plants in two regions in order to realise their global valorisation. Volatile extracts composition in the roots of Tussilago farfara L. and Calendula arvensis L., as well as flower buds of Spartium junceum L. were firstly investigated. The main chemical compounds in volatile extract from Tussilago farfara L. roots were sesquiterpene hydrocarbons and aliphatic compounds while main chemical compounds in volatile extract from Calendula arvensis L. roots were oxygenated sesquiterpenes, oxygenated monoterpenes and oxygenated diterpenes. The volatile extract from flower buds of Spartium junceum L. was mainly composed of aliphatic compounds. Antioxidant capacity evaluation results (by DPPH, ABTS, FRAC, ORAC and Folin-Ciocalteu tests) showed that several plant samples like Cytisus scoparius L., Tussilago farfara L., Citrum aurantium L. and Robinia pseudoacacia L. could be potential sources of natural antioxidants. Comparisons of hydrodistillation (HD), steam distillation (SD) and subcritical water extraction (SWE) showed that HD and SD had limited effects on essential oil composition but HD, SD and SWE had significant impacts on the recovery of antioxidants. Hydrodistillation seemed to be a better method for recovery of antioxidant compounds from residues of distillation than steam distillation. However, SWE appeared to be a more efficient method for direct extraction of antioxidant molecules (or phenolic compounds) from plants. In the hydrodistillation process, mineral contents in water were found to have very limited effects on yields of extracts but calcium and bicarbonate ions, had significant decreasing effects on antioxidant capacity and total phenolic content of both aqueous and methanolic extracts. Finally, an improved MAP-refinery was developed. Subcritical water was used for further extraction of antioxidant compounds from residues in original MAP-refinery. In this way, five parts could be obtained from plant materials: volatile extract, aqueous extract, methanolic extract, subcritical water extract and the final residue. The results showed that the improved MAP-refinery significantly increased the recovery of antioxidants compared with original MAP-refinery. This promising process will also allow a better valorisation of the final solid residue due to the lower content of residual water.