Yip Mei-kuen. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2000. / Includes bibliographical references (leaves 93-102). / Abstracts in English and Chinese. / Acknowledgments --- p.i / List of abbreviations --- p.ii / Abstract --- p.iii / Table of Contents --- p.vi / List of tables --- p.x / List of figures --- p.xii / Chapter 1. --- Introduction --- p.1 / Chapter 1.1 --- Plant of interest --- p.1 / Chapter 1.2 --- Literature review --- p.2 / Chapter 1.2.1 --- Anthocyanins-natural pigments in plants --- p.2 / Chapter 1.2.1.1 --- Sources and biosynthesis --- p.2 / Chapter 1.2.1.2 --- Chemical properties --- p.2 / Chapter 1.2.1.3 --- Biological effects --- p.3 / Chapter 1.2.2 --- Characterization of anthocyanins --- p.4 / Chapter 1.2.3 --- Plant tissue and cell cultures --- p.6 / Chapter 1.2.4 --- Induction of anthocyanins in plant tissue culture --- p.7 / Chapter 1.2.5 --- Factors affecting anthocyanin production --- p.8 / Chapter 1.2.5.1 --- Plant hormones --- p.8 / Chapter 1.2.5.2 --- Nutrients --- p.9 / Chapter 1.2.5.2.1 --- Phosphate --- p.9 / Chapter 1.2.5.2.2 --- Nitrogen --- p.9 / Chapter 1.2.5.3 --- Osmoticums --- p.10 / Chapter 1.2.5.3.1 --- Sucrose --- p.10 / Chapter 1.2.5.3.2 --- Other factors --- p.10 / Chapter 1.3 --- Research objectives --- p.12 / Chapter 2. --- Materials and methods --- p.16 / Chapter 2.1 --- Plant materials --- p.16 / Chapter 2.2 --- Study of pigment formation at different developmental stages --- p.16 / Chapter 2.2.1 --- Cultivation of Vigna sinensis --- p.16 / Chapter 2.2.2 --- Sample collection --- p.16 / Chapter 2.2.3 --- HPLC analysis of pigmented vegetative tissues --- p.16 / Chapter 2.2.4 --- HPLC analysis of seed coats at different developmental stages --- p.17 / Chapter 2.3 --- Characterization of seed coat pigments --- p.17 / Chapter 2.3.1 --- Extraction of seed coats pigments --- p.17 / Chapter 2.3.2 --- Acid hydrolysis of anthocyanins --- p.17 / Chapter 2.3.3 --- High performance liquid chromatography --- p.18 / Chapter 2.3.3.1 --- HPLC system --- p.18 / Chapter 2.3.3.2 --- Analytical conditions --- p.18 / Chapter 2.4 --- Establishment of tissue culture system --- p.19 / Chapter 2.4.1 --- Aseptic plant stocks --- p.19 / Chapter 2.4.2 --- Shoot-tip cultures --- p.19 / Chapter 2.4.3 --- Callus initiation --- p.19 / Chapter 2.4.3.1 --- From seed coats --- p.20 / Chapter 2.4.3.2 --- From vegetative tissues --- p.20 / Chapter 2.4.3.3 --- Light and dark --- p.20 / Chapter 2.4.4 --- Optimization of callus growth --- p.21 / Chapter 2.4.4.1 --- Basal medium --- p.21 / Chapter 2.4.4.2 --- Combination of various plant hormones --- p.21 / Chapter 2.4.4.3 --- Basal salt --- p.21 / Chapter 2.5 --- Studies of anthocyanin production in hypocotyl callus cultures --- p.22 / Chapter 2.5.1 --- Effects of nutrients --- p.22 / Chapter 2.5.1.1 --- Nitrogen --- p.22 / Chapter 2.5.1.2 --- Phosphate --- p.22 / Chapter 2.5.2 --- Osmotic stress --- p.22 / Chapter 2.5.2.1 --- Sucrose --- p.22 / Chapter 2.5.2.2 --- Mannitol --- p.23 / Chapter 2.5.2.3 --- Sodium chloride --- p.23 / Chapter 2.5.2.4 --- Polyethylene glycol --- p.23 / Chapter 2.6 --- Studies of anthocyanin production in cell suspension cultures --- p.23 / Chapter 2.6.1 --- Effects of nutrients --- p.24 / Chapter 2.6.1.1 --- Nitrogen --- p.24 / Chapter 2.6.1.2 --- Phosphate --- p.24 / Chapter 2.6.2 --- Osmotic stress --- p.25 / Chapter 2.6.2.1 --- Sucrose --- p.25 / Chapter 2.6.2.2 --- Polyethylene glycol --- p.25 / Chapter 2.6.3 --- Effects of other factors --- p.25 / Chapter 2.6.3.1 --- Riboflavin --- p.25 / Chapter 2.6.3.2 --- pH --- p.26 / Chapter 2.7 --- Measurement of cell growth --- p.26 / Chapter 2.8 --- Estimation of anthocyanins --- p.26 / Chapter 2.9 --- Statistical analysis --- p.27 / Chapter 3. --- Results --- p.30 / Chapter 3.1 --- Study of pigment formation at different developmental stages --- p.30 / Chapter 3.1.1 --- General description --- p.30 / Chapter 3.1.2 --- HPLC analysis of developing seed coats and other vegetative tissues --- p.30 / Chapter 3.1.3 --- The relationship between pigment formation and seed development --- p.30 / Chapter 3.2 --- Characterization of seed coat pigments --- p.31 / Chapter 3.3 --- Establishment of tissue culture system --- p.43 / Chapter 3.3.1 --- Callus initiations from seed coats --- p.43 / Chapter 3.3.2 --- Callus initiations from vegetative tissues --- p.43 / Chapter 3.3.3 --- Optimization of callus growth --- p.43 / Chapter 3.3.3.1 --- Effects of NAA and BA --- p.43 / Chapter 3.3.3.2 --- Effects of basal medium and combinations of plant hormones --- p.44 / Chapter 3.3.3.3 --- Effects of basal salt --- p.44 / Chapter 3.4 --- Studies of anthocyanin production in hypocotyl callus culture --- p.54 / Chapter 3.4.1 --- Effects of nutrients --- p.54 / Chapter 3.4.1.1 --- Effects of total nitrogen --- p.54 / Chapter 3.4.1.2 --- Effects of phosphate --- p.54 / Chapter 3.4.2 --- Effects of plant hormones --- p.55 / Chapter 3.4.3 --- Osmotic stress --- p.55 / Chapter 3.5 --- Establishment of suspension culture system --- p.64 / Chapter 3.6 --- Studies of anthocyanin production in seed coat suspension cultures --- p.64 / Chapter 3.6.1 --- Nutrient effects on suspension cultures --- p.64 / Chapter 3.6.2 --- Osmotic stress on suspension cultures --- p.65 / Chapter 3.6.3 --- Effects of phosphate with high nitrogen --- p.65 / Chapter 3.6.4 --- Effects of riboflavin with high nitrogen --- p.66 / Chapter 3.6.5 --- Influence of pH with high nitrogen --- p.66 / Chapter 4. --- Discussion --- p.79 / Chapter 4.1 --- Anthocyanin in vegetative tissues and seed coats of Vigna sinensis --- p.79 / Chapter 4.2 --- Factors affecting callus initiation in Vigna sinensis --- p.81 / Chapter 4.2.1 --- Explant types --- p.81 / Chapter 4.2.2 --- Plant hormones --- p.82 / Chapter 4.2.3 --- Basal medium --- p.82 / Chapter 4.3 --- Factors affecting anthocyanin production in callus cultures derived from hypocotyls --- p.83 / Chapter 4.3.1 --- Nutrients --- p.83 / Chapter 4.3.2 --- Osmotic stress --- p.85 / Chapter 4.4 --- Factors affecting anthocyanin production in suspension culture derived from seed coats --- p.86 / Chapter 4.4.1 --- Nutrients --- p.86 / Chapter 4.4.2 --- Osmotic stress --- p.87 / Chapter 4.5 --- Comparison of anthocyanin production from natural source and plant tissue cultures of V.sinensis --- p.89 / Chapter 4.6 --- Further studies --- p.89 / Chapter 5. --- Conclusions --- p.91 / References --- p.93

Lau Wing-Sze. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 131-149). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstract --- p.ii / Abstract (Chinese version) --- p.iv / Table of Contents --- p.vi / List of Tables --- p.x / List of Figures --- p.xii / List of Abbreviations --- p.xiv / Chapter Chapter One: --- An overview of Vigna sinensis seeds / Chapter 1.1 --- Introduction --- p.1 / Chapter 1.2 --- Food and functional food --- p.2 / Chapter 1.3 --- Edible legumes as an important food --- p.4 / Chapter 1.4 --- Nutritional an extra-nutritional values of V. sinensis seeds --- p.5 / Chapter Chapter Two: --- Purification of phenolic antioxidants from V. sinensis seeds / Chapter 2.1 --- Introduction --- p.11 / Chapter 2.1.1 --- Reactive oxygen species and antioxidants --- p.12 / Chapter 2.1.2 --- Phenolic flavonoids --- p.15 / Chapter 2.2 --- Materials and Methods / Chapter 2.2.1 --- Chemicals and reagents --- p.24 / Chapter 2.2.2 --- Plant material --- p.25 / Chapter 2.2.3 --- Optimization and extraction of V. sinensis seeds constituents --- p.25 / Chapter 2.2.4 --- Chromatographic separation of phenolic components --- p.26 / Chapter 2.2.5 --- Determination of phenolic contents --- p.27 / Chapter 2.2.6 --- Determination of free radical scavenging ability using trolox equivalent antioxidant capacity (TEAC) assay --- p.28 / Chapter 2.2.7 --- Statistical analysis --- p.29 / Chapter 2.3 --- Results and Discussion / Chapter 2.3.1 --- Optimization on the extraction of V. sinensis seeds --- p.30 / Chapter 2.3.2 --- Extraction and fractionation of V. sinensis seeds constituents --- p.31 / Chapter 2.3.3 --- Yield of various V sinensis seed fractions --- p.31 / Chapter 2.3.4 --- Phenolic contents in various V. sinensis seed fractions --- p.32 / Chapter 2.3.5 --- Free radical scavenging abilities of various V sinensis seed fractions --- p.33 / Chapter Chapter Three: --- Effect of V. sinensis seed extract on high fat and cholesterol - feeding mice / Chapter 3.1 --- Introduction --- p.41 / Chapter 3.1.1 --- Cholesterol in bloodstream circulation --- p.42 / Chapter 3.1.2 --- "Relationship between LDL oxidation, atherosclerosis and coronary heart disease" --- p.43 / Chapter 3.1.3 --- Diet supplements with beneficial effects on preventing coronary heart disease --- p.44 / Chapter 3.2 --- Materials and Methods --- p.47 / Chapter 3.2.1 --- Chemicals and reagents --- p.47 / Chapter 3.2.2 --- Preparation of diets --- p.48 / Chapter 3.2.3 --- Animals --- p.48 / Chapter 3.2.4 --- Feeding experiments --- p.49 / Chapter 3.2.5 --- Post-feeding analysis --- p.50 / Chapter 3.2.5.1 --- Caecal content and health indices --- p.50 / Chapter 3.2.5.2 --- Serum triglycerides --- p.51 / Chapter 3.2.5.3 --- Serum total cholesterol --- p.52 / Chapter 3.2.5.4 --- High-density lipoprotein (HDL) cholesterol --- p.53 / Chapter 3.2.5.5 --- Low-density lipoprotein (LDL) cholesterol --- p.54 / Chapter 3.2.5.6 --- Hepatic lipid and cholesterol --- p.55 / Chapter 3.2.6 --- Statistical analysis --- p.55 / Chapter 3.3 --- Results and Discussion --- p.56 / Chapter 3.3.1 --- Food intakes and body weights of animals --- p.56 / Chapter 3.3.2 --- Caecal contents and health indices --- p.56 / Chapter 3.3.3 --- Effects of V sinensis seed extract on serum and hepatic levels of triglycerides and cholesterol --- p.57 / Chapter Chapter Four: --- Antiproliferative activities of V. sinensis seed extracts / Chapter 4.1 --- Introduction --- p.66 / Chapter 4.1.1 --- Cancer and antioxidant --- p.67 / Chapter 4.1.2 --- Dietary cancer prevention agents --- p.68 / Chapter 4.2 --- Materials and Methods --- p.71 / Chapter 4.2.1 --- Chemicals and reagents --- p.71 / Chapter 4.2.2 --- Cell lines --- p.71 / Chapter 4.2.3 --- Maintenance of cell lines --- p.72 / Chapter 4.2.4 --- Antiproliferation assays --- p.73 / Chapter 4.2.4.1 --- MTT assay --- p.73 / Chapter 4.2.4.2 --- BrdU assay --- p.73 / Chapter 4.2.5 --- Cytotoxic activity determined by lactate dehydrogenase (LDH) assay --- p.77 / Chapter 4.2.6 --- Time-course assay --- p.79 / Chapter 4.2.7 --- Determination of IC50 --- p.79 / Chapter 4.2.8 --- Statistical analysis --- p.79 / Chapter 4.3 --- Results and Discussion --- p.80 / Chapter 4.3.1 --- Antiproliferative activities of V. sinensis seed extracts on HepG2 cells --- p.80 / Chapter 4.3.2 --- Cytotoxic activities of V. sinensis seed extracts on HepG2 cells --- p.82 / Chapter 4.3.3 --- Antiproliferative activities of phenolic fraction on MCF-7cells --- p.83 / Chapter 4.3.4 --- Cytotoxic activity of phenolic fraction on MCF-7 cells --- p.84 / Chapter 4.3.5 --- Time-course study of antiproliferative activities of phenolic fraction on cancer cells --- p.85 / Chapter 4.3.6 --- Effect of phenolic fraction on normal cells --- p.86 / Chapter Chapter Five: --- Antioxidant and antiproliferative activities of selected content flavonoids from V. sinensis seeds / Chapter 5.1 --- Introduction --- p.93 / Chapter 5.1.1 --- Cell cycle progression and regulation --- p.94 / Chapter 5.1.2 --- Bioavailability of plant flavonoids --- p.96 / Chapter 5.1.3 --- Characterization of flavonoids in V. sinensis seed --- p.98 / Chapter 5.2 --- Materials and Methods --- p.102 / Chapter 5.2.1 --- Chemicals and reagents --- p.102 / Chapter 5.2.2 --- Determination of free radical scavenging ability of identified flavonoids from V sinensis seeds using trolox equivalent antioxidant capacity (TEAC) assay --- p.103 / Chapter 5.2.3 --- Antiproliferation assays --- p.104 / Chapter 5.2.4 --- Cytotoxicity assay --- p.104 / Chapter 5.2.5 --- Time-course assay --- p.104 / Chapter 5.2.6 --- Determination of cell cycle distribution by flow cytometry --- p.105 / Chapter 5.2.7 --- Statistical analysis --- p.106 / Chapter 5.3 --- Results and Discussion --- p.107 / Chapter 5.3.1 --- Free radical scavenging abilities of identified flavonoids from V sinensis seeds --- p.107 / Chapter 5.3.2 --- Antiproliferative activities of selected flavonoids on cancer cells --- p.109 / Chapter 5.3.3 --- Cytotoxic activities of selected flavonoids on cancer cells --- p.111 / Chapter 5.3.4 --- Time -course study of antiproliferative activities on cancer cells --- p.112 / Chapter 5.3.5 --- Cytotoxic activities of selected flavonoids on normal cells --- p.114 / Chapter 5.3.6 --- Determination of the effects of cyanidin on cancer cells by analyzing cell cycle pattern --- p.115 / Chapter Chapter Six: --- Conclusion --- p.128 / References --- p.131

by Foo Wai Ting, Rita. / Thesis submitted in: August 1998. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1999. / Includes bibliographical references (leaves 89-100). / Abstract also in Chinese. / Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Proximate Composition --- p.4 / Chapter 1.2 --- Amino Acid Composition --- p.6 / Chapter 1.3 --- Antinutrients --- p.11 / Chapter 1.3.1 --- Trypsin Inhibitors --- p.12 / Chapter 1.3.2 --- Phytate --- p.13 / Chapter 1.3.3 --- Tannins --- p.14 / Chapter 1.3.4 --- Lectins --- p.15 / Chapter 1.4 --- Two Dimensional Polyacrylamide Gel Electrophoresis --- p.17 / Chapter 1.5 --- Protein Digestibility --- p.19 / Chapter 1.6 --- Protein Quality --- p.22 / Chapter 1.7 --- Hypocholesterolemic Effects --- p.24 / Chapter 2 --- Materials and Methods --- p.36 / Chapter 2.1 --- Plant Material --- p.36 / Chapter 2.2 --- Sample preparation --- p.36 / Chapter 2.3 --- Proximate composition --- p.38 / Chapter 2.3.1 --- Protein --- p.38 / Chapter 2.3.2 --- Fat --- p.38 / Chapter 2.3.3 --- Carbohydrate --- p.38 / Chapter 2.3.4 --- Fiber --- p.38 / Chapter 2.3.5 --- Mineral --- p.39 / Chapter 2.3.6 --- Moisture --- p.39 / Chapter 2.4 --- Amino acid composition --- p.40 / Chapter 2.5 --- Antinutrients --- p.41 / Chapter 2.5.1 --- Trypsin inhibitors --- p.41 / Chapter 2.5.2 --- Tannins --- p.42 / Chapter 2.5.3 --- Phytate --- p.43 / Chapter 2.5.4 --- Lectins --- p.43 / Chapter 2.6 --- Two dimensional polyacrylamide gel electrophoresis --- p.45 / Chapter 2.6.1 --- Protein extraction --- p.45 / Chapter 2.6.2 --- IEF gel --- p.45 / Chapter 2.6.3 --- SDS gel --- p.46 / Chapter 2.7 --- Protein digestibility --- p.48 / Chapter 2.7.1 --- In vitro Protein digestibility --- p.48 / Chapter 2.7.2 --- True Protein digestibility --- p.49 / Chapter 2.8 --- Protein quality --- p.51 / Chapter 2.9 --- Hypocholesterolemic effects --- p.52 / Chapter 2.10 --- Statistical analysis --- p.55 / Chapter 3 --- Results --- p.56 / Chapter 3.1 --- Proximate composition --- p.56 / Chapter 3.2 --- Amino acid composition --- p.56 / Chapter 3.3 --- Antinutrients --- p.56 / Chapter 3.4 --- Two dimensional polyacrylamide gel electrophoresis --- p.60 / Chapter 3.5 --- Protein digestibility --- p.60 / Chapter 3.6 --- Protein quality --- p.60 / Chapter 3.7 --- Hypocholesterolemic effects --- p.62 / Chapter 3.7.1 --- Growth rate against day --- p.62 / Chapter 3.7.2 --- Health indexes --- p.64 / Chapter 3.7.3 --- Cholesterol content --- p.64 / Chapter 4 --- Discussion --- p.67 / Chapter 4.1 --- Proximate composition --- p.67 / Chapter 4.2 --- Amino acid composition --- p.70 / Chapter 4.3 --- Antinutrients --- p.74 / Chapter 4.4 --- Two dimensional polyacrylamide gel electrophoresis --- p.77 / Chapter 4.5 --- Protein digestibility --- p.79 / Chapter 4.6 --- Protein quality --- p.81 / Chapter 4.7 --- Hypocholesterolemic effects --- p.82 / Chapter 5 --- Conclusion --- p.88 / References --- p.89

Vochysiaceae compreende uma família com duas tribos, oito gêneros e aproximadamente 250 espécies distribuídas predominantemente na América Tropical. Vochysia pyramidalis e V. tucanorum são espécies arbóreas, a primeira distribuída nas regiões nordeste e central do Brasil e a última apresentando distribuição semelhante, mas alcançando latitudes meridionais até o sul do Paraná. Estas e outras espécies de Vochysiaceae apresentam teores relativamente altos de lipídeos de sementes. Os lipídeos de sementes de V. pyramidalis assemelham-se aos da manteiga de cacau, com altos teores de ácido esteárico e ácido oleico, e os de V. tucanorum são ricos em ácidos graxos de cadeia longa, como os ácidos erúcico e docosanóico, assemelhando-se ao óleo original de colza. Os lipídeos de ambas as espécies têm, portanto, potencial valor econômico. Foi verificado que as proporções dos ácidos graxos das duas espécies podem variar entre populações de diferentes localidades. A proposta do presente trabalho foi detectar variações na distribuição química das duas espécies, a fim de apontar populações com lipídeos dotados de perfis de ácidos graxos mais convenientes para potencial uso medicinal e/ou industrial, e verificar possível conexão entre perfis químicos e genéticos. O último foi estabelecido com base em marcadores AFLP (Amplified Fragment Length Polymorphism). Foram coletadas sementes e fragmentos de folhas de 3-9 indivíduos de V. pyramidalis de populações de Alto Paraíso de Goiás (GO), Andaraí (BA), Palmeiras (BA) e UnB-Brasília (DF), e 5-10 indivíduos de V. tucanorum de populações de Bauru (SP), Botucatu (SP), Santana do Riacho (MG), Perdões (MG), IBGE-Brasília (DF), Ibicoara (BA) e Sengés (PR). Os lipídeos das sementes foram extraídos com hexano. Foram seguidos métodos padronizados para a obtenção dos correspondentes ésteres metílicos dos ácidos graxos. Estes foram analisados por CG/FID e a identificação das substâncias foram baseadas na comparação dos tempos de retenção com aqueles de amostras autênticas de ésteres metílicos de ácidos graxos. A distribuição dos ácidos graxos das duas espécies foi analisada pelo coeficiente de Distância Euclidiana Simples e pelo método de agrupamento UPGMA e Análise de Componente Principal (PCA), usando o programa de computador Fitopac 1.6.4. Dois agrupamentos principais foram obtidos, cada um correspondendo a uma das espécies de Vochysiaceae. Entretanto, foi notada uma alta semelhança entre amostras da mesma espécie. Em cada espécie, não foi obtido nenhum agrupamento coerente relacionado à populações e localidades. A única exceção correspondeu aos indivíduos da população de V. pyramidalis de Alto Paraíso de Goiás, que se agrupou isoladamente de outras amostras da mesma espécie. Análises de AFLP foram realizadas com DNA extraído de fragmentos de folhas preservados em silicagel, seguidos de procedimentos padronizados de digestão, ligação, amplificações pré-seletiva e seletiva, e análise em seqüenciador automático. Três e quatro combinações de iniciadores foram usadas para V. pyramidalis e V. tucanorum, respectivamente. Os fragmentos obtidos foram utilizados como caracteres e analisados pelo método de evolução mínima Neighbor-Joining, usando distância de Nei & Li por meio do programa de computador PAUP v. 4.0, e Análise de Coordenada Principal (PCO) usando o programa de computador Fitopac 1.6.4. Assim como ocorreu com os caracteres químicos, pequenas distâncias foram obtidas ao se comparar amostras de cada uma das duas espécies de Vochysia. Em análises individuais de cada espécie, não foi obtido nenhum agrupamento coerente com as respectivas populações. Os resultados do presente trabalho sugerem que a dispersão de indivíduos de V. pyramidalis e V. tucanorum nas presentes áreas de distribuição é um evento recente, não tendo decorrido tempo suficiente para alcançar diferenças químicas e genéticas detectáveis. Essa observação é coerente com estudos da dispersão de sementes e de pólen, que mostrou uma alta capacidade de dispersão de espécies de Vochysia. / The Vochysiaceae comprise a family with two tribes, eight genera and approximately 250 species distributed predominantly in tropical America. Vochysia pyramidalis and V. tucanorum are tree species, the former distributed in Central, Northeastern and Southeasten Brazil and the latter having similar distribution, but reaching meridional latitudes as far as the south of Paraná. These two species and other Vochysiaceae have been shown to have relatively high levels of seeds lipids. The seed lipids of V. pyramidalis resemble cocoa butter, with high levels of stearic and oleic acids, while V. tucanorum seeds are rich in long chain fatty acids, such as erucic and docosanoic, resembling the original rapeseed oil. Hence, lipids of both species of Vochysiaceae have potential economic value. It has been reported that proportions of seed fatty acids of the two species may vary among populations of different localities. The aim of the present work was to detect variations in chemical profiles of the two species, in order to point out populations with lipids endowed with fatty acid profiles more convenient for potential medical and/or industrial uses, and verify possible connection between chemical and genetic profiles. The latter was established on basis of AFLP (amplified fragment length polymorphism) markers. Seeds and leaf fragments were collected of 3-9 individuals of V. pyramidalis from populations of Alto Paraíso de Goiás (GO), Andaraí (BA), Palmeiras (BA) and UnB-Brasília (DF) and 5-10 of V. tucanorum from populations of Bauru (SP), Botucatu (SP), Santana do Riacho (MG), Perdões (MG), IBGE-Brasília (DF), Ibicoara (BA) and Sengés(PR). Seed lipids were extracted with hexane. Standardized methods were followed to obtain the corresponding fatty acids methyl esters. These were analyzed by GC/FID and identification of the substances were based on comparison of retention times with those of authentic samples of fatty acid methyl esters. The distribution of fatty acids of the samples of the two species was analyzed by Simple Euclidean Distance coefficients, clusthering method UPGMA and Principal Component Analysis (PCA) using Fitopac 1.6.4 as computer program. Two main clusters were obtained, each corresponding to one of the Vochysiaceae species. However, a high similarity was noted among samples of the same species. Within each species, no clusters were obtained coherent with populations and localities. The only exception corresponded to individuals of a population of V. pyramidalis of Alto Paraíso de Goiás, which grouped apart from other samples of the same species. AFLP analyses were carried out with DNA extracted from leaf fragments preserved in silicagel, following the standardized procedures of digestion, ligation, pre and selective amplifications and analysis in automatic sequencer. Three and four primer combinations were used regarding V. pyramidalis and V. tucanorum, respectively. Fragments obtained were scored as characters and analyzed by the Neighbor-Joining method, using Nei & Li distances in a computer program implemented in PAUP v. 4.0 and Principal Coordinate Analysis (PCO) using Fitopac 1.6.4 as computer program. As with chemical characters, small distances were obtained comparing samples of each the two Vochysia species. In the analyses of individuals of each species, no clusters were obtained coherently with the respective population. The results of the present work suggest that the dispersion of individuals of V. pyramidalis and V. tucanorum into the present areas of distribution is a recent event, no enough time having been elapsed to reach detectable chemical and genetic differences. This observation is coherent with studies of seed and polen dispersal, which have shown a high disperal capacity of Vochysia species.

Vochysiaceae compreende uma família com duas tribos, oito gêneros e aproximadamente 250 espécies distribuídas predominantemente na América Tropical. Vochysia pyramidalis e V. tucanorum são espécies arbóreas, a primeira distribuída nas regiões nordeste e central do Brasil e a última apresentando distribuição semelhante, mas alcançando latitudes meridionais até o sul do Paraná. Estas e outras espécies de Vochysiaceae apresentam teores relativamente altos de lipídeos de sementes. Os lipídeos de sementes de V. pyramidalis assemelham-se aos da manteiga de cacau, com altos teores de ácido esteárico e ácido oleico, e os de V. tucanorum são ricos em ácidos graxos de cadeia longa, como os ácidos erúcico e docosanóico, assemelhando-se ao óleo original de colza. Os lipídeos de ambas as espécies têm, portanto, potencial valor econômico. Foi verificado que as proporções dos ácidos graxos das duas espécies podem variar entre populações de diferentes localidades. A proposta do presente trabalho foi detectar variações na distribuição química das duas espécies, a fim de apontar populações com lipídeos dotados de perfis de ácidos graxos mais convenientes para potencial uso medicinal e/ou industrial, e verificar possível conexão entre perfis químicos e genéticos. O último foi estabelecido com base em marcadores AFLP (Amplified Fragment Length Polymorphism). Foram coletadas sementes e fragmentos de folhas de 3-9 indivíduos de V. pyramidalis de populações de Alto Paraíso de Goiás (GO), Andaraí (BA), Palmeiras (BA) e UnB-Brasília (DF), e 5-10 indivíduos de V. tucanorum de populações de Bauru (SP), Botucatu (SP), Santana do Riacho (MG), Perdões (MG), IBGE-Brasília (DF), Ibicoara (BA) e Sengés (PR). Os lipídeos das sementes foram extraídos com hexano. Foram seguidos métodos padronizados para a obtenção dos correspondentes ésteres metílicos dos ácidos graxos. Estes foram analisados por CG/FID e a identificação das substâncias foram baseadas na comparação dos tempos de retenção com aqueles de amostras autênticas de ésteres metílicos de ácidos graxos. A distribuição dos ácidos graxos das duas espécies foi analisada pelo coeficiente de Distância Euclidiana Simples e pelo método de agrupamento UPGMA e Análise de Componente Principal (PCA), usando o programa de computador Fitopac 1.6.4. Dois agrupamentos principais foram obtidos, cada um correspondendo a uma das espécies de Vochysiaceae. Entretanto, foi notada uma alta semelhança entre amostras da mesma espécie. Em cada espécie, não foi obtido nenhum agrupamento coerente relacionado à populações e localidades. A única exceção correspondeu aos indivíduos da população de V. pyramidalis de Alto Paraíso de Goiás, que se agrupou isoladamente de outras amostras da mesma espécie. Análises de AFLP foram realizadas com DNA extraído de fragmentos de folhas preservados em silicagel, seguidos de procedimentos padronizados de digestão, ligação, amplificações pré-seletiva e seletiva, e análise em seqüenciador automático. Três e quatro combinações de iniciadores foram usadas para V. pyramidalis e V. tucanorum, respectivamente. Os fragmentos obtidos foram utilizados como caracteres e analisados pelo método de evolução mínima Neighbor-Joining, usando distância de Nei & Li por meio do programa de computador PAUP v. 4.0, e Análise de Coordenada Principal (PCO) usando o programa de computador Fitopac 1.6.4. Assim como ocorreu com os caracteres químicos, pequenas distâncias foram obtidas ao se comparar amostras de cada uma das duas espécies de Vochysia. Em análises individuais de cada espécie, não foi obtido nenhum agrupamento coerente com as respectivas populações. Os resultados do presente trabalho sugerem que a dispersão de indivíduos de V. pyramidalis e V. tucanorum nas presentes áreas de distribuição é um evento recente, não tendo decorrido tempo suficiente para alcançar diferenças químicas e genéticas detectáveis. Essa observação é coerente com estudos da dispersão de sementes e de pólen, que mostrou uma alta capacidade de dispersão de espécies de Vochysia. / The Vochysiaceae comprise a family with two tribes, eight genera and approximately 250 species distributed predominantly in tropical America. Vochysia pyramidalis and V. tucanorum are tree species, the former distributed in Central, Northeastern and Southeasten Brazil and the latter having similar distribution, but reaching meridional latitudes as far as the south of Paraná. These two species and other Vochysiaceae have been shown to have relatively high levels of seeds lipids. The seed lipids of V. pyramidalis resemble cocoa butter, with high levels of stearic and oleic acids, while V. tucanorum seeds are rich in long chain fatty acids, such as erucic and docosanoic, resembling the original rapeseed oil. Hence, lipids of both species of Vochysiaceae have potential economic value. It has been reported that proportions of seed fatty acids of the two species may vary among populations of different localities. The aim of the present work was to detect variations in chemical profiles of the two species, in order to point out populations with lipids endowed with fatty acid profiles more convenient for potential medical and/or industrial uses, and verify possible connection between chemical and genetic profiles. The latter was established on basis of AFLP (amplified fragment length polymorphism) markers. Seeds and leaf fragments were collected of 3-9 individuals of V. pyramidalis from populations of Alto Paraíso de Goiás (GO), Andaraí (BA), Palmeiras (BA) and UnB-Brasília (DF) and 5-10 of V. tucanorum from populations of Bauru (SP), Botucatu (SP), Santana do Riacho (MG), Perdões (MG), IBGE-Brasília (DF), Ibicoara (BA) and Sengés(PR). Seed lipids were extracted with hexane. Standardized methods were followed to obtain the corresponding fatty acids methyl esters. These were analyzed by GC/FID and identification of the substances were based on comparison of retention times with those of authentic samples of fatty acid methyl esters. The distribution of fatty acids of the samples of the two species was analyzed by Simple Euclidean Distance coefficients, clusthering method UPGMA and Principal Component Analysis (PCA) using Fitopac 1.6.4 as computer program. Two main clusters were obtained, each corresponding to one of the Vochysiaceae species. However, a high similarity was noted among samples of the same species. Within each species, no clusters were obtained coherent with populations and localities. The only exception corresponded to individuals of a population of V. pyramidalis of Alto Paraíso de Goiás, which grouped apart from other samples of the same species. AFLP analyses were carried out with DNA extracted from leaf fragments preserved in silicagel, following the standardized procedures of digestion, ligation, pre and selective amplifications and analysis in automatic sequencer. Three and four primer combinations were used regarding V. pyramidalis and V. tucanorum, respectively. Fragments obtained were scored as characters and analyzed by the Neighbor-Joining method, using Nei & Li distances in a computer program implemented in PAUP v. 4.0 and Principal Coordinate Analysis (PCO) using Fitopac 1.6.4 as computer program. As with chemical characters, small distances were obtained comparing samples of each the two Vochysia species. In the analyses of individuals of each species, no clusters were obtained coherently with the respective population. The results of the present work suggest that the dispersion of individuals of V. pyramidalis and V. tucanorum into the present areas of distribution is a recent event, no enough time having been elapsed to reach detectable chemical and genetic differences. This observation is coherent with studies of seed and polen dispersal, which have shown a high disperal capacity of Vochysia species.