Increased temperatures due to anthropogenic-induced climate change may raise the threat of extinction for taxa with sessile life histories (e.g., plants) in the near future. Linking climate change models to demographic models may provide useful insights into the potential effects of environmental changes on rare plants, and therefore aid in their current and future conservation. Population demographers generally agree that mechanistic models from a reductionist perspective are necessary to test assumptions in population drivers. For the first study, I assessed the climate vulnerability of a rare plant species, Pyrrocoma radiata, with a mechanistic model of four climatically-similar populations. I used environmentally-driven demographic models to estimate vital rates and population sizes from a nonlinear, nonparametric regression with local climate variables. I assessed the utility of this environmentally-correlated, stage-structured population matrix model compared to a stationary model of independent and identically-distributed environmental stochasticity. I then simulated future population projections based on climate conditions predicted by General Circulation Models (GCMs) under opposing emission scenarios. The second study hopes to answer population-level questions using a traditionally community-level method, non-metric multidimensional scaling, which considers correlation structure between response variables and can be used to find environmental correlates of the ordination axes. Demographic data on a threatened perennial, Astragalus tyghensis, were collected from five sites in the Tygh Valley, OR. I considered correlation structure between demographic vital rates to find environmental correlates of the ordination axes. The search for an environmental driver of population vital rates was successful for the two study species. Previous year dry dormant season precipitation likely affects the fertility rates a year later in P. radiata populations, and dry growing season reference evapotranspiration rates positively correlated with a growth gradient in A. tyghensis. Based on predicted precipitation, P. radiata is expected to rapidly decline by 2050, but this may be due to biases in the two GCMs and reliance on only one environmental factor. The NMS ordination adequately captured most of the variation in transition elements for the years and populations from A. tyghensis demographics. I provided support to the claim that model predictions can improve with the inclusion of mechanistic relationships. The inclusion of abiotic drivers in models used to predict population trends is supported by our study and may enhance predictive power in population viability assessments under changing climates. / Graduation date: 2013

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No. of bitstreams: 2 dissertacao_tuane_araldi_da_silva.pdf: 1228072 bytes, checksum: 53d0b042633ad80a28404e1dc2bc54db (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2015-02-27 / Sem bolsa / Os trabalhos foram realizados no Laboratório de Fisiologia de Sementes - UFPel, nos anos de 2012 a 2014 e objetivaram avaliar o efeito de diferentes concentrações de extratos vegetais sobre aspectos fisiológicos de sementes e plântulas de alface e trigo. Nos capítulos 1 e 2 foram utilizados extratos de folhas de buva (Conyza bonariensis) e azevém (Lolium multiflorum)nas concentrações 0; 2; 4; 6 e 8% e como espécie alvo foram utilizadas sementes de alface. No capítulo 3, a espécie alvo foi trigo e o extrato aquoso utilizado foi de folhas de Lolium multiflorum nas concentrações 0; 2; 4; 6 e 8%. Foram avaliados a germinação, a primeira contagem de germinação, o índice de velocidade de germinação, o comprimento de parte aérea e de raiz primária, a massa seca total de plântulas, a condutividade elétrica, a atividade das enzimas superóxido-dismutase (SOD), catalase (CAT) e ascorbarto-peroxidase (APX), peroxidação lipídica, peróxido de hidrogênio, emergência de plântulas, área foliar, comprimento de parte aérea e de raiz e massa seca total das plântulas emergidas. No capítulo 1, verificou-se que as maiores concentrações do extrato aquoso de buva proporcionaram maior inibição da germinação e do crescimento inicial de plântulas de alface. Houve aumento da condutividade elétrica em sementes de alface, níveis de peróxido de hidrogênio, da peroxidação lipídica e atividade das enzimas superóxido-dismutase, catalase, ascorbato-peroxidase. No capítulo 2 verificou-se que o extrato aquoso de folhas de azevém afetou negativamente a primeira contagem de germinação, o índice de velocidade de germinação e o crescimento de plântulas de alface. O aumento da concentração do extrato proporcionou a elevação na atividade das enzimas antioxidantes, sendo resultados mais pronunciados, observados nas concentrações de 6 e 8%. No capítulo 3, o índice de velocidade de germinação de sementes de trigo foi reduzido de forma marcante na concentração 8% do extrato de azevém. As concentrações mais altas do extrato resultaram nos maiores teores de peróxido de hidrogênio, peroxidação lipídica e atividades das enzimas superóxido-dismutase, catalase e ascorbato-peroxidase. / The works was carried out at the Seed Laboratory of Physiology - UFPel in the years 2012 to 2014 and aimed to evaluate the effect of different concentrations of plant extracts on physiological aspects of seeds and lettuce and wheat seedlings. In chapters 1 and 2 leaf extracts were used horseweed (Conyza bonariensis) and ryegrass (Lolium multiflorum) at concentrations of 0; 2; 4; 6 to 8% as the target species and lettuce seeds were used. In Chapter 3, the target species was wheat and the aqueous extract was of Lolium multiflorum leaves at concentrations of 0; 2; 4; 6 to 8%. We evaluated the germination, the first count, the germination rate index, shoot length and primary root, total seedling dry weight, electrical conductivity, the activity of superoxide dismutase (SOD), catalase (CAT) and ascorbarto-peroxidase (APX), lipid peroxidation, hydrogen peroxide, seedling emergence, leaf area, shoot and root length and total dry mass of emerged seedlings. In Chapter 1, it was found that higher concentrations of the aqueous extract showed higher horseweed inhibition of germination and initial growth of lettuce seedlings. There was an increase of the electrical conductivity lettuce seeds, hydrogen peroxide levels, lipid peroxidation and activity of superoxide dismutase, catalase, ascorbate peroxidase. In Chapter 2 it was found that the aqueous extract of ryegrass leaves negatively affected the first count, the germination speed index and the growth of lettuce seedlings. The increase of the extract concentration resulted in higher in antioxidant enzyme activity, being more pronounced results observed at concentrations of 6 to 8%. In Chapter 3, the wheat seed germination speed was reduced markedly in concentration 8% of ryegrass extract. Higher concentrations of the extract produced the highest yields of hydrogen peroxide, lipid peroxidation and activities of superoxide dismutase, catalase and ascorbate peroxidase.

O presente trabalho refere-se ao desenvolvimento de grânulos a partir de produto seco de A. satureioides obtido por spray drying, em escala semi-industrial, visando a superar as principais limitações tecnológicas apresentadas tais como a higroscopia excessiva do produto, baixa densidade, pequeno tamanho de partícula e instabilidade dos constituintes flavonoídicos frente à luz. Dois métodos foram empregados para a granulação: granulação em leito fluidizado e granulação via seca por desagregação. Estes métodos apresentaram, respectivamente, 15 e 60 % de rendimento do processo, levando a escolha da granulação seca para a produção dos grânulos do produto seco. A granulação seca resultou em grânulos assimétricos, de faixa granulométrica situada entre 0,3 mm e 1,9 mm e de superfície irregular e rugosa. O revestimento destes grânulos foi realizado em leito fluidizado, utilizando três tipos de polímeros, Eudragit® L 30D, Opadry® II e Opadry® AMB.O rendimento médio com o Opadry® II foi de 72 %. O revestimento com Opadry® AMB foi realizado utilizando-se um baixo e alto fluxo do líquido de revestimento, obtendo-se rendimentos de 75 e 85 %, respectivamente. A higroscopia dos grânulos foi avaliada em ambientes com umidade relativa controlada de 65 ou 99 %, comparando-se o comportamento dos grânulos revestidos com o dos grânulos sem a presença de revestimento. Ambos os filmes com Opadry® não protegeram os grânulos frente à umidade. As fotomicrografias por MEV, destes grânulos, revelaram que, ambos os tipos de revestimento, apresentaram superfícies rugosas e com presença de poros explicando, parcialmente, a falta de proteção contra a umidade. O estudo da fotoproteção indica, em um primeiro momento, que o filme de revestimento promove a proteção dos flavonóides frente a ação da luz. A avaliação preliminar da liberação dos flavonóides, a partir dos grânulos, foi realizada em células de fluxo Desaga® com os grânulos sem revestimento e revestidos com Opadry® AMB. A água não demonstrou ser o meio mais favorável para este ensaio. Em meio com pH 1,2 contendo 1 % de laurilsulfato de sódio, os flavonóides quercetina, luteolina e 3-Ometilquercetina foram liberados, respectivamente, após 90 minutos, em 70, 83 e 70 % a partir dos grânulos não revestidos, e 52, 54 e 45 % a partir dos grânulos revestidos com Opadry® AMB, denotando a influência do filme de revestimento sobre o perfil de liberação dos flavonóides. Em seu conjunto, os resultados obtidos nesta primeira abordagem deste tema abrem diversas perspectivas para oaprofundamento do estudo e desenvolvimento de granulados de A. satureioides, a partir de produto seco por spray drying. / The present work was designed to develop granules from Achyrocline satureioides spray dried powders in order to overcome the main technological limitations presented by this herbal raw material: high hygroscopy, low density, small particle diameter and flavonoid sensibility against light. Two methods were employed for granulation, fluidized bed and dry granulation. These methods presented, respectively, 15 and 60 % of process yield, leading to the selection of dry granulation for preparing the granules. By this method, the granules showed 0,3 to 1,9 mm particle mean diameter, rough surface and irregular morphology. For coating the granules, three types of polymers were employed, Eudragit® L30D, Opadry® II and Opadry® AMB, in fluidized bed. The first one resulted in atomizer obstruction becoming non viable its employment. The mean yield coating obtained with Opadry® II was 72 %. The Opadry® AMB was sprayed in low and high rates yielding, respectively, 75 % and 85 %. The hygroscopicity of the granules was evaluated in controlled conditions of 65 % or 99 % RH, comparing the behavior of the coated granules which that presented by non-coated particles. Both Opadry® coatings did not protect the granules against the humidity. The SEM photomicrographis of these granules revealed that both types of coating presented rough and porous surface explaining partially, at least, the lack of protection against the humidity. The preliminary photostability assay indicated, in a first view, that the coating promotes the flavonoid protection. The flavonoid release from the granules was performed in Desaga® flow cell from the non-coated granules and from Opadry® AMB coated granules. Water demonstrated not to be an appropriate media for both tested granules. In pH 1.2 media containing 1 % of sodium lauryl sulfate, the flavonoids quercetin, luteolin and 3-O-methylquercetin were, respectively, released, after 90 minutes, in 70 %, 83 % and 70 % from non coated granules and in 52 %, 54 % and 45 % from Opadry® AMB coated granules, denoting the influence of the coating on the flavonoid release profile. Taken together, the results of this first approach openmay perspectives for more detailed studies on Achyrocline satureioides granules from the corresponding spray dried powders.

O presente trabalho refere-se ao desenvolvimento de grânulos a partir de produto seco de A. satureioides obtido por spray drying, em escala semi-industrial, visando a superar as principais limitações tecnológicas apresentadas tais como a higroscopia excessiva do produto, baixa densidade, pequeno tamanho de partícula e instabilidade dos constituintes flavonoídicos frente à luz. Dois métodos foram empregados para a granulação: granulação em leito fluidizado e granulação via seca por desagregação. Estes métodos apresentaram, respectivamente, 15 e 60 % de rendimento do processo, levando a escolha da granulação seca para a produção dos grânulos do produto seco. A granulação seca resultou em grânulos assimétricos, de faixa granulométrica situada entre 0,3 mm e 1,9 mm e de superfície irregular e rugosa. O revestimento destes grânulos foi realizado em leito fluidizado, utilizando três tipos de polímeros, Eudragit® L 30D, Opadry® II e Opadry® AMB.O rendimento médio com o Opadry® II foi de 72 %. O revestimento com Opadry® AMB foi realizado utilizando-se um baixo e alto fluxo do líquido de revestimento, obtendo-se rendimentos de 75 e 85 %, respectivamente. A higroscopia dos grânulos foi avaliada em ambientes com umidade relativa controlada de 65 ou 99 %, comparando-se o comportamento dos grânulos revestidos com o dos grânulos sem a presença de revestimento. Ambos os filmes com Opadry® não protegeram os grânulos frente à umidade. As fotomicrografias por MEV, destes grânulos, revelaram que, ambos os tipos de revestimento, apresentaram superfícies rugosas e com presença de poros explicando, parcialmente, a falta de proteção contra a umidade. O estudo da fotoproteção indica, em um primeiro momento, que o filme de revestimento promove a proteção dos flavonóides frente a ação da luz. A avaliação preliminar da liberação dos flavonóides, a partir dos grânulos, foi realizada em células de fluxo Desaga® com os grânulos sem revestimento e revestidos com Opadry® AMB. A água não demonstrou ser o meio mais favorável para este ensaio. Em meio com pH 1,2 contendo 1 % de laurilsulfato de sódio, os flavonóides quercetina, luteolina e 3-Ometilquercetina foram liberados, respectivamente, após 90 minutos, em 70, 83 e 70 % a partir dos grânulos não revestidos, e 52, 54 e 45 % a partir dos grânulos revestidos com Opadry® AMB, denotando a influência do filme de revestimento sobre o perfil de liberação dos flavonóides. Em seu conjunto, os resultados obtidos nesta primeira abordagem deste tema abrem diversas perspectivas para oaprofundamento do estudo e desenvolvimento de granulados de A. satureioides, a partir de produto seco por spray drying. / The present work was designed to develop granules from Achyrocline satureioides spray dried powders in order to overcome the main technological limitations presented by this herbal raw material: high hygroscopy, low density, small particle diameter and flavonoid sensibility against light. Two methods were employed for granulation, fluidized bed and dry granulation. These methods presented, respectively, 15 and 60 % of process yield, leading to the selection of dry granulation for preparing the granules. By this method, the granules showed 0,3 to 1,9 mm particle mean diameter, rough surface and irregular morphology. For coating the granules, three types of polymers were employed, Eudragit® L30D, Opadry® II and Opadry® AMB, in fluidized bed. The first one resulted in atomizer obstruction becoming non viable its employment. The mean yield coating obtained with Opadry® II was 72 %. The Opadry® AMB was sprayed in low and high rates yielding, respectively, 75 % and 85 %. The hygroscopicity of the granules was evaluated in controlled conditions of 65 % or 99 % RH, comparing the behavior of the coated granules which that presented by non-coated particles. Both Opadry® coatings did not protect the granules against the humidity. The SEM photomicrographis of these granules revealed that both types of coating presented rough and porous surface explaining partially, at least, the lack of protection against the humidity. The preliminary photostability assay indicated, in a first view, that the coating promotes the flavonoid protection. The flavonoid release from the granules was performed in Desaga® flow cell from the non-coated granules and from Opadry® AMB coated granules. Water demonstrated not to be an appropriate media for both tested granules. In pH 1.2 media containing 1 % of sodium lauryl sulfate, the flavonoids quercetin, luteolin and 3-O-methylquercetin were, respectively, released, after 90 minutes, in 70 %, 83 % and 70 % from non coated granules and in 52 %, 54 % and 45 % from Opadry® AMB coated granules, denoting the influence of the coating on the flavonoid release profile. Taken together, the results of this first approach openmay perspectives for more detailed studies on Achyrocline satureioides granules from the corresponding spray dried powders.

O presente trabalho refere-se ao desenvolvimento de grânulos a partir de produto seco de A. satureioides obtido por spray drying, em escala semi-industrial, visando a superar as principais limitações tecnológicas apresentadas tais como a higroscopia excessiva do produto, baixa densidade, pequeno tamanho de partícula e instabilidade dos constituintes flavonoídicos frente à luz. Dois métodos foram empregados para a granulação: granulação em leito fluidizado e granulação via seca por desagregação. Estes métodos apresentaram, respectivamente, 15 e 60 % de rendimento do processo, levando a escolha da granulação seca para a produção dos grânulos do produto seco. A granulação seca resultou em grânulos assimétricos, de faixa granulométrica situada entre 0,3 mm e 1,9 mm e de superfície irregular e rugosa. O revestimento destes grânulos foi realizado em leito fluidizado, utilizando três tipos de polímeros, Eudragit® L 30D, Opadry® II e Opadry® AMB.O rendimento médio com o Opadry® II foi de 72 %. O revestimento com Opadry® AMB foi realizado utilizando-se um baixo e alto fluxo do líquido de revestimento, obtendo-se rendimentos de 75 e 85 %, respectivamente. A higroscopia dos grânulos foi avaliada em ambientes com umidade relativa controlada de 65 ou 99 %, comparando-se o comportamento dos grânulos revestidos com o dos grânulos sem a presença de revestimento. Ambos os filmes com Opadry® não protegeram os grânulos frente à umidade. As fotomicrografias por MEV, destes grânulos, revelaram que, ambos os tipos de revestimento, apresentaram superfícies rugosas e com presença de poros explicando, parcialmente, a falta de proteção contra a umidade. O estudo da fotoproteção indica, em um primeiro momento, que o filme de revestimento promove a proteção dos flavonóides frente a ação da luz. A avaliação preliminar da liberação dos flavonóides, a partir dos grânulos, foi realizada em células de fluxo Desaga® com os grânulos sem revestimento e revestidos com Opadry® AMB. A água não demonstrou ser o meio mais favorável para este ensaio. Em meio com pH 1,2 contendo 1 % de laurilsulfato de sódio, os flavonóides quercetina, luteolina e 3-Ometilquercetina foram liberados, respectivamente, após 90 minutos, em 70, 83 e 70 % a partir dos grânulos não revestidos, e 52, 54 e 45 % a partir dos grânulos revestidos com Opadry® AMB, denotando a influência do filme de revestimento sobre o perfil de liberação dos flavonóides. Em seu conjunto, os resultados obtidos nesta primeira abordagem deste tema abrem diversas perspectivas para oaprofundamento do estudo e desenvolvimento de granulados de A. satureioides, a partir de produto seco por spray drying. / The present work was designed to develop granules from Achyrocline satureioides spray dried powders in order to overcome the main technological limitations presented by this herbal raw material: high hygroscopy, low density, small particle diameter and flavonoid sensibility against light. Two methods were employed for granulation, fluidized bed and dry granulation. These methods presented, respectively, 15 and 60 % of process yield, leading to the selection of dry granulation for preparing the granules. By this method, the granules showed 0,3 to 1,9 mm particle mean diameter, rough surface and irregular morphology. For coating the granules, three types of polymers were employed, Eudragit® L30D, Opadry® II and Opadry® AMB, in fluidized bed. The first one resulted in atomizer obstruction becoming non viable its employment. The mean yield coating obtained with Opadry® II was 72 %. The Opadry® AMB was sprayed in low and high rates yielding, respectively, 75 % and 85 %. The hygroscopicity of the granules was evaluated in controlled conditions of 65 % or 99 % RH, comparing the behavior of the coated granules which that presented by non-coated particles. Both Opadry® coatings did not protect the granules against the humidity. The SEM photomicrographis of these granules revealed that both types of coating presented rough and porous surface explaining partially, at least, the lack of protection against the humidity. The preliminary photostability assay indicated, in a first view, that the coating promotes the flavonoid protection. The flavonoid release from the granules was performed in Desaga® flow cell from the non-coated granules and from Opadry® AMB coated granules. Water demonstrated not to be an appropriate media for both tested granules. In pH 1.2 media containing 1 % of sodium lauryl sulfate, the flavonoids quercetin, luteolin and 3-O-methylquercetin were, respectively, released, after 90 minutes, in 70 %, 83 % and 70 % from non coated granules and in 52 %, 54 % and 45 % from Opadry® AMB coated granules, denoting the influence of the coating on the flavonoid release profile. Taken together, the results of this first approach openmay perspectives for more detailed studies on Achyrocline satureioides granules from the corresponding spray dried powders.

Please read the abstract in the section 07chapter7 of this document / Thesis (DPhil (Plant Physiology))--University of Pretoria, 2007. / Plant Science / unrestricted

Der Ursprung, die Entwicklung und die Formierung von Laubblatt-Mustern konnten bei ausgewählten variegaten Formen der Araceae, Asteraceae, Ericaceae, Maranthaceae und Rosaceae erklärt werden. Die Pflanzen wurden je nach Problematik untersucht und in drei verschiedene Gruppen verteilt: In der ersten Gruppe, Blattmuster mit unregelmäßiger makulater Musterung, Monstera deliciosa, Syngonium podophyllum und die Sorten 'Pirol' und 'Luyona' von Dendranthema grandiflorum zeigen ein unregelmäßiges Laubblattmuster und keine weiß-randigen bzw. weißkernigen Periklinalchimären. Mischzellen wurden durch direkte (mikroskopisch) und indirekte (In-vitro-Kultur und Selbstungen) Nach der Plastidenentmischung in den Schichten des Sprossmeristems wurde bei Syngonium, Monstera und den zwei Sorten von Dendranthema die GW-Form als einzige stabile Periklinalchimäre nachgewiesen. In der zweiten Gruppe, Immerspaltende Periklinalchimären, die chimärische Konstitution GA bei Spiraea bumalda 'Goldflame' konnte durch Wurzelaustriebe (BATESON-Test) und die Adventivsprossinduktion aus Kallus nachgewiesen werden. Darüber hinaus konnten mehrfach perikline Aufspaltungen der ersten Sprossscheitelschicht (Reduplikation von L1) nachgewiesen werden, die zur Entstehung der Panaschierung von 'Goldflame' zur Entmischung führten. Bei den Untersuchungen der In-vitro-Regenerate aus der Kalluskultur und der Wurzelaustriebe an S. bumalda 'Shirobana' wurde festgestellt, dass diese Pflanze keine Chimäre ist und das auftretende Muster der Blüten genetisch kontrolliert ist. In der dritten Gruppe, Hypoderm und Beeinflussung der Musterbildung: Die unmaskierten Binnenfelder bei Ctenanthe lubbersiana 'Variegata' und der Rhododendron-Hybride 'Goldflimmer' sind durch die Existenz eines Hypoderms zu erklären. Bei Ctenanthe lubbersiana 'Variegata' befindet sich ein Hypoderm an der Blattoberseite und der Blattunterseite. Bei 'Goldflimmer' liegt nur unter der oberen Epidermis ein einschichtiges Hypoderm vor. Infolgedessen fehlt an der Blattoberseite die Maskierung Das gelbe Binnenfeld des Blattmusters ist durch eine grüne Mesophyllschicht unterlagert. / The origin, development and formation of foliage-leaf-patterns could be explained with selected variegaten forms of the Araceae, Asteraceae, Ericaceae, Marantaceae and Rosaceae. In order to prove this the plants were examined according to the problem and classified in three different groups: In the first group, leaf-patterns with irregular maculated patterns, Monstera deliciosa, Syngonium podophyllum and the sorts 'Pirol' and 'Luyona' of Dendranthema grandiflorum showed an irregular foliage-leaf-pattern, thought neither to show periclinal chimeras with white edges nor with green edges. Mixed cells were detected by direct (microscopic) and indirect (In vitro culture and self pollination) test. After the plastid sorting out in the layers of the meristems, the green over white Form was proven with Syngonium, Monstera and the two sorts of Dendranthema as a single stable periclinal chimera. In the second group, eversporting periclinal chimeras, the green over aurea chimeral constitution with Spiraea bumalda 'Goldflame' was proved by the regeneration of adventitious shoots from their roots (BATESON-Test) and also by the induction of adventitious bud from callus. Periclinal divisions of the first layer of meristems (reduplication of L1), which are responsible for the appearing of green pattern of the leafs was proved many times. Examination of the regenerated shoots from callus and from the adventitious shoots from roots of S. bumalda 'Shirobana' showed that this plant is not a chimera and that the appearing pattern of the blooms is genetically controlled. In the third group, hypoderm and influence of the pattern-formation, the unmasked inner-fields with Ctenanthe lubbersiana 'Variegata' and the Rhododendron-hybrid 'Goldflimmer' were explained through the existence of one layered hypoderm under the upper Epidermis as well as over the lower Epidermis of C. lubbersiana 'Variegata', thought in 'Goldflimmer' it is only found a one layer Hypoderm under the upper epidermis. Subsequently, the masking is missing at the upper-leaf side the yellow inner-field of the leaf-pattern is through a green Mesophyllslayer masked.

Shum Ka Chiu. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (leaves 174-182). / Abstracts in English and Chinese. / Acknowledgement --- p.ii / Abstract --- p.iii / 摘要 --- p.vi / Table of content --- p.viii / List of figures --- p.xvi / List of tables --- p.xxii / Abbreviations --- p.xxv / Chapter Chapter 1. --- Introduction --- p.1 / Chapter 1.1 --- Importance of authentication of Traditional Chinese Medicines --- p.1 / Chapter 1.1.1 --- Confusing nomenclatures --- p.1 / Chapter 1.1.2 --- Similar morphologies of different medicinal materials --- p.2 / Chapter 1.1.3 --- Toxicities of medicinal materials --- p.2 / Chapter 1.1.4 --- Conservation of natural products --- p.2 / Chapter 1.2 --- TCM listed in the Pharmacopoeia of People's Republic of China --- p.3 / Chapter 1.3 --- Overview of mis-use and intoxication of TCM --- p.4 / Chapter 1.4 --- Ordinances regulating Chinese medicines as natural products --- p.7 / Chapter 1.4.1 --- Laws governing Chinese medicine --- p.7 / Chapter 1.4.2 --- Laws governing endangered species --- p.8 / Chapter 1.5 --- Current technologies in the authentication of Traditional Chinese Medicines and their limitations --- p.9 / Chapter 1.6 --- Historical applications of Radix Aconiti --- p.12 / Chapter 1.7 --- Modern applications of Radix Aconiti --- p.16 / Chapter 1.8 --- Research on Radix Aconiti and its chemical components --- p.17 / Chapter 1.8.1 --- Chemistry --- p.17 / Chapter 1.8.2 --- Pharmacology --- p.19 / Chapter 1.8.3 --- Molecular interaction --- p.22 / Chapter 1.9 --- Brief review on the systematics and phylogeny of Aconitum --- p.23 / Chapter 1.10 --- Historical applications of Radix Aucklandiae and related materials --- p.25 / Chapter 1.11 --- Modern applications of Radix Aucklandiae and related material --- p.27 / Chapter 1.12 --- Research on Aucklandiae and related material and their chemical components --- p.28 / Chapter 1.12.1 --- Chemistry --- p.28 / Chapter 1.12.2 --- Pharmacology --- p.29 / Chapter 1.13 --- Brief review on the systematics and phylogeny of Aucklandia and related medicinal species --- p.31 / Chapter 1.14 --- Authentication by DNA sequencing --- p.33 / Chapter 1.14.1 --- Introduction --- p.33 / Chapter 1.14.2 --- Criteria of sequence markers --- p.36 / Chapter 1.14.3 --- Model used to process polymorphism in DNA sequences --- p.37 / Chapter 1.15 --- Screening for novel markers --- p.38 / Chapter 1.15.1 --- Reason for screening novel markers --- p.38 / Chapter 1.15.2 --- Basic principle --- p.39 / Chapter 1.16 --- Introduction to gas chromatography- mass spectrometry --- p.40 / Chapter 1.16.1 --- Basic principles and components of GC-MS --- p.41 / Chapter 1.16.2 --- Advantages and limitations of GC-MS --- p.42 / Chapter 1.16.3 --- Usage of GC-MS on natural product analysis --- p.43 / Chapter 1.16.4 --- Chemometric analysis --- p.44 / Chapter 1.17 --- Objectives --- p.46 / Chapter Chapter 2. --- Materials and Methods --- p.47 / Chapter 2.1 --- Plant samples --- p.47 / Chapter 2.1.1 --- Samples of Aconitum --- p.47 / Chapter 2.1.2 --- Samples of Aucklandia and related species --- p.51 / Chapter 2.2 --- DNA extraction method --- p.58 / Chapter 2.2.1 --- Reagents --- p.58 / Chapter 2.2.2 --- Methods --- p.59 / Chapter 2.3 --- Chemical extraction methods --- p.61 / Chapter 2.4 --- Chemical standard extraction and purification method --- p.62 / Chapter 2.5 --- DNA sequencing --- p.63 / Chapter 2.5.1 --- Reagents --- p.63 / Chapter 2.5.2 --- Methods --- p.65 / Chapter 2.6 --- Genomic subtraction --- p.70 / Chapter 2.7 --- Search for species-specific markers from the subtraction library --- p.74 / Chapter 2.8 --- Gas chromatography- mass spectrometry --- p.74 / Chapter 2.9 --- GC-MS chemometric analysis --- p.75 / Chapter Chapter 3. --- Authentication of Aconitum by DNA Sequencing --- p.76 / Chapter 3.1 --- Introduction --- p.76 / Chapter 3.2 --- Methods --- p.77 / Chapter 3.3 --- Results - 5S spacer --- p.77 / Chapter 3.3.1 --- Sequence information --- p.77 / Chapter 3.3.2 --- Sequence similarity --- p.78 / Chapter 3.3.3 --- Phylogram study --- p.81 / Chapter 3.4 --- Results -psbA-trnH --- p.85 / Chapter 3.4.1 --- Sequence information --- p.85 / Chapter 3.4.2 --- Sequence similarity --- p.85 / Chapter 3.4.3 --- Phylogram study --- p.87 / Chapter 3.5 --- Discussion --- p.91 / Chapter 3.5.1 --- Overview of nuclear ribosomal 5S spacer --- p.91 / Chapter 3.5.2 --- Extensive polymorphism of 5S spacer --- p.91 / Chapter 3.5.3 --- Distribution of samples in the phylograms constructed by 5S spacer --- p.93 / Chapter 3.5.4 --- Utility of 5S spacer for authentication --- p.94 / Chapter 3.5.5 --- Overview of psbA-trnH spacer --- p.94 / Chapter 3.5.6 --- Distribution of samples in the phylograms constructed by psbA-trnH spacer --- p.95 / Chapter 3.5.7 --- A distinctive region of inversion --- p.96 / Chapter 3.5.8 --- Utility of psbA-trnH for authentication --- p.97 / Chapter Chapter 4. --- Screening for Novel Markers for Authentication of Aconitum --- p.98 / Chapter 4.1 --- Introduction --- p.98 / Chapter 4.2 --- Methods --- p.99 / Chapter 4.3 --- Results - subtracted clones --- p.99 / Chapter 4.4 --- Results - SSH6 --- p.104 / Chapter 4.4.1 --- Sequence information --- p.104 / Chapter 4.4.2 --- Sequence similarity --- p.105 / Chapter 4.5 --- Results-SSH15 --- p.107 / Chapter 4.5.1 --- Sequence information --- p.107 / Chapter 4.5.2 --- Sequence similarity --- p.107 / Chapter 4.5.3 --- Phylogram study --- p.109 / Chapter 4.6 --- Results-SSH45 --- p.113 / Chapter 4.6.1 --- Sequence information --- p.113 / Chapter 4.6.2 --- Sequence similarity --- p.113 / Chapter 4.6.3 --- Phylogram study --- p.115 / Chapter 4.7 --- Discussion --- p.119 / Chapter 4.7.1 --- Utility of subtraction in screening markers --- p.119 / Chapter 4.7.2 --- SSH6 --- p.121 / Chapter 4.7.3 --- SSH15 --- p.122 / Chapter 4.7.4 --- SSH45 --- p.123 / Chapter 4.7.5 --- Hybridization in Aconitum --- p.124 / Chapter 4.7.6 --- Inferring species identities of samples from the market --- p.126 / Chapter 4.8 --- Conclusion --- p.128 / Chapter Chapter 5. --- Assessment of Aucklandia lappa and Related Species by GC-MS --- p.129 / Chapter 5.1 --- Introduction --- p.129 / Chapter 5.2 --- Methods --- p.130 / Chapter 5.3 --- Results --- p.130 / Chapter 5.3.1 --- Extraction of essential oil --- p.130 / Chapter 5.3.2 --- GC-MS analysis --- p.131 / Chapter 5.3.3 --- Peak alignment and hierarchical cluster analysis --- p.133 / Chapter 5.3.4 --- Purification of chemical markers from Aucklandia lappa --- p.148 / Chapter 5.3.5 --- Standardization of the purified chemical markers --- p.148 / Chapter 5.3.6 --- Quantitative analysis of chemical markers --- p.152 / Chapter 5.4 --- Discussion --- p.154 / Chapter 5.4.1 --- Analysis of chemical composition --- p.154 / Chapter 5.4.2 --- A comparison on chemometric methods --- p.154 / Chapter 5.4.3 --- Similarity of chemical profiles --- p.156 / Chapter 5.4.4 --- Dendrogram analysis --- p.157 / Chapter 5.4.5 --- Utility of GC-MS in authentication of A. lappa and related species --- p.159 / Chapter 5.4.6 --- Limitations --- p.159 / Chapter 5.4.7 --- Comparison with molecular data --- p.161 / Chapter 5.4.8 --- Contents of dehydrocostuslactone and costunolide --- p.163 / Chapter 5.4.9 --- Locality study --- p.164 / Chapter 5.5 --- Conclusion --- p.165 / Chapter Chapter 6. --- General Discussion --- p.167 / Chapter 6.1 --- DNA sequencing --- p.168 / Chapter 6.2 --- Genomic subtraction --- p.169 / Chapter 6.3 --- Future work on molecular authentication --- p.170 / Chapter 6.4 --- Future work on authentication of Aconitum --- p.170 / Chapter 6.5 --- Gas chromatography- mass spectrometry --- p.171 / Chapter 6.6 --- Future work on authentication by GC-MS --- p.172 / Chapter 6.7 --- Future work on authentication of Aucklandia lappa and related species … --- p.173 / References --- p.174 / Appendix A. Sequence Alignment of 5S Spacer from Aconitum Species --- p.183 / Appendix B. Sequence Alignment of psbA- trnH Spacer from Aconitum Species --- p.188 / Appendix C. Sequences of Subtracted Clones from Aconitum --- p.191 / Appendix D. Sequence Alignment of SSH6 from Aconitum Species --- p.194 / Appendix E. Sequence Alignment of SSH15 from Aconitum Species --- p.195 / Appendix F. Sequence Alignment of SSH45 from Aconitum Species --- p.200 / Appendix G. Gas Chromatograms of Essential Oil Extracts of Aucklandia lappa and Related Species --- p.202