This thesis discusses the symbolism found in Walt Whitman's second poetic period, as found in the collection Leaves of Grass.

Ho Hing Man. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 140-156). / Abstracts in English and Chinese. / Chapter Chapter 1 --- General introduction / Chapter 1.1 --- History of soybean --- p.1 / Chapter 1.2 --- Health benefits of soybean --- p.2 / Chapter 1.3 --- Introduction to flavonoids --- p.2 / Chapter 1.4 --- Bioavailability of flavonoids from foods --- p.3 / Chapter 1.5 --- Pharmacological effects of flavonoids and their glycosides --- p.4 / Chapter 1.5.1 --- Anticarcinogenic activity --- p.4 / Chapter 1.5.2 --- Antioxidative activity --- p.7 / Chapter 1.5.3 --- Cardioprotective activity --- p.9 / Chapter 1.5.4 --- Osteoprotective activity --- p.10 / Chapter 1.5.5 --- Neuroprotective activity --- p.12 / Chapter 1.5.6 --- Antiangiogenic activity --- p.12 / Chapter 1.6 --- Soy leaves --- p.13 / Chapter Chapter 2 --- Isolation and purification of kaempferol glycosides and genistin in soy leaves / Chapter 2.1 --- Introduction --- p.14 / Chapter 2.2 --- Objectives --- p.15 / Chapter 2.3 --- Materials and Methods --- p.16 / Chapter 2.3.1 --- Extraction and isolation --- p.16 / Chapter 2.3.1.1 --- Preparation of soy leaves butanol extract --- p.16 / Chapter 2.3.1.2 --- Preparation of kaempferol glycosides from soy leaves butanol extract --- p.16 / Chapter 2.3.2 --- High performance liquid chromatography (HPLC) analysis --- p.19 / Chapter 2.3.2.1 --- Sample preparation for the HPLC analysis --- p.19 / Chapter 2.3.2.2 --- HPLC analysis --- p.19 / Chapter 2.3.2.3 --- Quantification of the flavonoids and their glycosides --- p.23 / Chapter 2.3.2.4 --- Change in flavonoids and their glycosides in soy leaves --- p.23 / Chapter 2.4 --- Results --- p.24 / Chapter 2.4.1 --- Compound 1 --- p.24 / Chapter 2.4.2 --- Compound 2 --- p.24 / Chapter 2.4.3 --- Compound 3 --- p.25 / Chapter 2.4.4 --- Compound 4 --- p.25 / Chapter 2.4.5 --- Compound 5 --- p.25 / Chapter 2.4.6 --- Compound 6 --- p.26 / Chapter 2.4.7 --- Quantification of flavonoids in soybean and soy leaves --- p.32 / Chapter 2.4.8 --- Age-dependent changes in flavonoids and their glycosides --- p.32 / Chapter 2.5 --- Discussion --- p.35 / Chapter 2.5.1 --- Compound 1 --- p.35 / Chapter 2.5.2 --- Compound 2 --- p.35 / Chapter 2.5.3 --- Compound 3 --- p.37 / Chapter 2.5.4 --- Compound 4 --- p.38 / Chapter 2.5.5 --- Compound 5 --- p.39 / Chapter 2.5.6 --- Compound 6 --- p.40 / Chapter 2.5.7 --- Age-dependent changes in flavonoids and their glycosides --- p.40 / Chapter Chapter 3 --- Hypolipidemic effects of soy leaves in hamsters / Chapter 3.1 --- Introduction --- p.41 / Chapter 3.1.1 --- Different lipoproteins and their functions --- p.41 / Chapter 3.1.2 --- Risk factors of cardiovascular disease --- p.42 / Chapter 3.1.3 --- Animal model --- p.43 / Chapter 3.2 --- Objectives --- p.44 / Chapter 3.3 --- Materials and Methods --- p.45 / Chapter 3.3.1 --- Animals --- p.46 / Chapter 3.3.2 --- Serum lipid and lipoprotein determinations --- p.46 / Chapter 3.3.3 --- Determination of cholesterol in the liver and adipose tissue --- p.46 / Chapter 3.3.4 --- Extraction of neutral and acidic sterols from fecal samples --- p.49 / Chapter 3.3.4.1 --- Determination of neutral sterols --- p.49 / Chapter 3.3.4.2 --- Determination of acidic sterols --- p.50 / Chapter 3.3.4.3 --- GLC analysis of neutral and acidic sterols --- p.51 / Chapter 3.3.5 --- Statistics --- p.51 / Chapter 3.4 --- Results --- p.54 / Chapter 3.4.1 --- Growth and food intake --- p.54 / Chapter 3.4.2 --- "Effects of SLP and SLEE supplementation on serum triacylglycerol (TG), total cholesterol (TC) and high-density lipoprotein cholesterol (HDL-C)" --- p.54 / Chapter 3.4.3 --- Effects ofSLP and SLEE supplementation on non-HDL-C and ratio of non-HDL-C to HDL-C --- p.55 / Chapter 3.4.4 --- Effects of SLP amd SLEE supplementations on concentration of hepatic cholesterol --- p.58 / Chapter 3.4.5 --- Effects of SLP and SLEE supplementations on perirenal adipose tissue cholesterol --- p.58 / Chapter 3.4.6 --- Effects of SLP and SLEE supplementations on fecal neutral and acidic sterols --- p.61 / Chapter 3.5 --- Discussion --- p.64 / Chapter Chapter 4 --- Effects of soy leaves and its flavonoid glycosides on haemolysis and on LDL oxidation / Chapter 4.1 --- Introduction --- p.67 / Chapter 4.1.1 --- Role of low density lipoprotein oxidation in the development of atherosclerosis --- p.68 / Chapter 4.1.2 --- LDL oxidation --- p.70 / Chapter 4.1.3 --- Thiobarbituric acid reactive substances (TBARS) as an index of LDL oxidation --- p.71 / Chapter 4.1.4 --- Antioxidant and LDL oxidation --- p.74 / Chapter 4.2 --- Objective --- p.75 / Chapter 4.3 --- Materials and methods --- p.76 / Chapter 4.3.1 --- Isolation of LDL from human serum --- p.76 / Chapter 4.3.2 --- LDL oxidation --- p.77 / Chapter 4.3.3 --- Determine the formation of thiobarbituric acid-reactive substances (TBARS) --- p.77 / Chapter 4.3.4 --- Assay for erythrocyte haemolysis --- p.78 / Chapter 4.3.5 --- Statistics --- p.79 / Chapter 4.4 --- Results --- p.80 / Chapter 4.4.1 --- Effects of three different soy leaves extracts and flavonoid glycosides on LDL oxidation --- p.80 / Chapter 4.4.2 --- Effects of three soy leaves extracts and flavonoid glycosides on erythrocyte haemolysis --- p.80 / Chapter 4.5 --- Discussion --- p.85 / Chapter Chapter 5 --- Relaxing effects of soy leaves and its flavonoids / Chapter 5.1 --- Introduction --- p.89 / Chapter 5.1.1 --- Smooth muscle contraction --- p.90 / Chapter 5.1.1.1 --- Sliding filament mechanism --- p.91 / Chapter 5.1.2 --- Intracellular mechanisms involved in the regulation of smooth muscle contraction --- p.92 / Chapter 5.1.2.1 --- Voltage-gated Ca2+ channels --- p.92 / Chapter 5.1.2.2 --- Protein kinase C (PKC) mediated smooth muscle contraction --- p.93 / Chapter 5.1.2.3 --- Thromboxane A2 receptor-mediated calcium channel --- p.94 / Chapter 5.2 --- Objectives --- p.96 / Chapter 5.3 --- Materials and methods --- p.97 / Chapter 5.3.1 --- Drugs preparation --- p.97 / Chapter 5.3.2 --- Vessel preparation --- p.97 / Chapter 5.3.3 --- Contraction experiments --- p.99 / Chapter 5.3.3.1 --- Relaxant responses of soy leaves butanol extract on the contraction induced by different constrictors --- p.99 / Chapter 5.3.3.2 --- Relaxant responses of soy leaves butanol extract on U46619 and PGF2a- induced contraction --- p.99 / Chapter 5.3.3.3 --- "Relaxant responses of genistein, genistin and the kaempferol glycosides on U46619-induced contraction" --- p.100 / Chapter 5.3.4 --- Statistics --- p.100 / Chapter 5.4 --- Results --- p.102 / Chapter 5.4.1 --- Effect of soy leaves butanol extract --- p.102 / Chapter 5.4.2 --- Role of endothelium in extract-induced relaxation --- p.102 / Chapter 5.4.3 --- Effect of the soy leaves butanol extract on contractile response to prostaglandins --- p.103 / Chapter 5.4.4 --- Effects of kaempferol glycosides and kaempferol --- p.111 / Chapter 5.4.5 --- Effects of genistein and genistin --- p.111 / Chapter 5.5 --- Discussion --- p.118 / Chapter Chapter 6 --- Effect of soy leaves on mammary tumor / Chapter 6.1 --- Introduction --- p.123 / Chapter 6.1.1 --- Carcinogenesis --- p.123 / Chapter 6.1.1.1 --- In itiation --- p.124 / Chapter 6.1.1.2 --- Promotion --- p.124 / Chapter 6.1.1.3 --- Progression --- p.125 / Chapter 6.2 --- Objective --- p.126 / Chapter 6.3 --- Materials and methods --- p.127 / Chapter 6.3.1 --- Animal --- p.127 / Chapter 6.3.2 --- Determination of estrus cycle --- p.128 / Chapter 6.3.3 --- Statistics --- p.129 / Chapter 6.4 --- Results --- p.131 / Chapter 6.4.1 --- Incident rate of tumor induction --- p.131 / Chapter 6.4.2 --- Number of tumor induced --- p.131 / Chapter 6.5 --- Discussion --- p.136 / Chapter Chapter 7 --- Conclusions --- p.136 / References --- p.140

Filmes à base de polímeros contendo compostos com atividade antioxidante, podem ser usados na produção de embalagem ativa, que interagem com o alimento contribuindo na sua conservação e no prolongamento da vida de prateleira de alimentos ricos em componentes sensíveis ao oxigênio. Os extratos vegetais são ricos em polifenóis, conhecidos por sua atividade antioxidante e são uma alternativa ao uso dos antioxidantes sintéticos. Assim, o objetivo principal desta tesa foi o desenvolvimento de filmes à base de gelatina adicionados de extratos vegetais, com propriedades físicas e funcionais para seu emprego como embalagens bioativas. Foram produzidos extratos aquosos de sementes de guaraná (Paullinia cupana Kunth), folha de amoreira preta (Morus nigra L.), carqueja (Baccharis trimera), hortelã (Mentha piperita) e folha de pitangueira (Eugenia uniflora L.). Os extratos produzidos em três temperaturas (30, 45 e 60 °C) foram caracterizados quanto a sua cor instrumental, teor de sólidos solúveis, matéria seca, pH, composição proximal e em relação a sua atividade antioxidante (capacidade redutora, DPPH e ABTS). Os extratos que apresentaram maiores atividades antioxidantes foram utilizados na produção dos filmes. Os filmes foram produzidos pela técnica casting com 8 g de gelatina/100g de solução e glicerol (30 g de glicerol/100 g de gelatina) como plastificante. A quantidade de extrato adicionada foi estudada (0, 50 e 150 g de extrato/100 g de gelatina) para a obtenção das melhores propriedades antioxidantes. Os filmes foram submetidos às seguintes caracterizações: espessura, umidade, solubilidade, cor, opacidade, brilho, permeabilidade ao vapor de água, propriedades mecânicas (tração e perfuração), microestrutura (espectroscopia de força atômica e microscopia eletrônica de varredura), propriedades térmicas (DSC), estrutura química com uso de espectroscopia no infravermelho com transformada de Fourier e determinação da atividade antioxidante (capacidade redutora, DPPH e ABTS). O estudo demonstrou que as amostras vegetais analisadas e as temperaturas de extração escolhidas foram eficientes na produção de extratos com atividade antioxidante, sendo possível sua utilização para produção de filmes ativos. Os extratos que apresentaram maiores atividades antioxidantes foram os extratos de folha de pitangueira, semente de guaraná e hortelã, extraídos a 60 °C, sendo estes escolhidos para a produção dos filmes. Para a caracterização dos filmes, a adição dos extratos em geral não afetou nas propriedades mecânicas (tração e perfuração), umidade e solubilidade, não existindo diferença significativa entre os filmes controles sem extratos e os com extratos. A cor dos filmes sofreu influência na adição dos extratos, sendo que filmes acrescidos com os extratos apresentaram uma tendência mais escura e amarelada que os filmes controles. Entretanto, observou-se que, como esperado, os filmes com extratos demonstraram um aumento na atividade antioxidante e propriedades de barreira, sendo os acrescidos com extrato de folha de pitangueira, os que apresentaram melhores resultados. / Films based on biopolymers containing compounds with antioxidant activity can be used to produce active packaging, which interact with the food, contributing to it conservation and the increasing of the shelf life of the food rich in compounds oxygen sensitive. Plant extracts are rich in polyphenols, known for their antioxidant capacity and are an alternative to the use of synthetic antioxidants. Thus, the general goal of this thesis was to develop gelatin-based films added with vegetal extracts, with physical and functional properties for their use as bioactive packaging. Liquid extracts of Guarana (Paullinia cupana Kunth) seeds, mulberry (Morus nigra L.) leaves carqueja (Baccharis trimera.), peppermint (Mentha piperita) and pitanga (Eugenia uniflora L.) leaves were produced. The extracts produced in three temperatures (30, 45 and 60 °C) were characterized by their antioxidant capacity (total phenols, DPPH and ABTS), instrumental color, soluble solids, dry matter, pH and proximal composition. The extracts with the highest antioxidant activity were used in the film\'s production. The films were produced by the casting technique with 8 g of gelatin / 100 g of solution and glycerol (30 g of glycerol/100 g of gelatin) as plasticizer. The amount of extract added to studied the antioxidant properties were : 0, 50 and 150 g of extract / 100 g of gelatin. The films were subjected to the following characterizations: thickness, mechanical properties (traction and perforation), color, opacity, gloss, humidity, solubility, water vapor permeability, microstructure (Atomic force spectroscopy and scanning electron microscopy), thermal properties (DSC), determination of the antioxidant capacity, determination of reducing capacity and chemical structure using Fourier transform infrared spectroscopy. The study demonstrated that the analyzed plant samples and the chosen extraction temperatures were efficient in the production of extracts with antioxidant activity, being possible its use for the production of active films. Extracts of pitanga leaves, guarana seed and peppermint, produced at 60 °C, were chosen due to their high antioxidant activity. In general, the characterization of the films shown that the addition of the extracts did not affect in the mechanical, solubility and humidity properties, having no significant difference between the control films, with no extract, and those with extracts. The colour of the films were influenced by the addiction of the extract, with the increase of darkness and yellowness of the films, when films were added with the extracts. As expected, it was observed that the films with extracts showed an increase in the antioxidant capacity and barrier properties, being that films added with extracts of pitanga leaves presented the better results.

Development in multicellular organisms relies on establishing and maintaining gene expression profiles that give cells identity. Transcription factors establish gene expression profiles by integrating positional, temporal, and environmental cues to regulate genes essential for a cell's identity. These signals are often short lived while the differentiated state may persist for a long time. Epigenetic factors maintain these gene expression profiles by making heritable chemical alterations to target gene chromatin to stabilize transcriptional patterns. Here we explore the evolution and function of an epigenetic regulator, the Polycomb Repressive Complex 2 (PRC2), and a transcription factor, ASYMMETRIC LEAVES 1 (AS1) , in the lower eudicot Aquilegia. PRC2 is an important and deeply conserved epigenetic regulator, which is critical to many plant developmental processes, including the regulation of major developmental transitions and lateral organ development. We find that Aquilegia has a relatively simple complement of PRC2 genes that are expressed throughout development. Contrary to findings in other plant species, two members of the Aquilegia PRC2, AqSWN and AqCLF, are not imprinted in Aquilegia endosperm. Using virusinduced gene silencing (VIGS), we determined that Aquilegia PRC2 regulates aspects of lateral organ development, including branching within the leaf and lamina expansion, along with caroteinoid production in floral organs. PRC2 targeting of several floral MADS box genes may be conserved in Aquilegia, but other known targets such as the class I KNOX gene are not. AS1 is a transcription factor that plays a conserved role in controlling differentiation and polarity of lateral organs. In species with simple leaves, AS1 promotes cell determination by suppressing the expression of the class I KNOX genes in leaf primordia and regulates abaxial-adaxial polarity in the developing leaf. However, in species with compound leaves, KNOX genes and AS1 often work together to control leaflet initiation and arrangement. In Aquilegia, AqAS1 appears to primarily contribute to proper regulation of class I KNOX genes with a more minor role in leaflet polarity and positioning. Most interestingly, these combined datasets suggest that contrary to the widely held model, class I KNOX genes are neither necessary nor sufficient for leaf complexity in Aquilegia.

Thesis (doctoral)--Swedish University of Agricultural Sciences, 2005. / Thesis documentation sheet inserted. Appendix consists of reproductions of 5 published papers and manuscripts, four co-authored with others. Includes bibliographical references. Issued also electronically via World Wide Web in PDF format; online version lacks appendix of papers; map in online version displays in color.

Filmes à base de polímeros contendo compostos com atividade antioxidante, podem ser usados na produção de embalagem ativa, que interagem com o alimento contribuindo na sua conservação e no prolongamento da vida de prateleira de alimentos ricos em componentes sensíveis ao oxigênio. Os extratos vegetais são ricos em polifenóis, conhecidos por sua atividade antioxidante e são uma alternativa ao uso dos antioxidantes sintéticos. Assim, o objetivo principal desta tesa foi o desenvolvimento de filmes à base de gelatina adicionados de extratos vegetais, com propriedades físicas e funcionais para seu emprego como embalagens bioativas. Foram produzidos extratos aquosos de sementes de guaraná (Paullinia cupana Kunth), folha de amoreira preta (Morus nigra L.), carqueja (Baccharis trimera), hortelã (Mentha piperita) e folha de pitangueira (Eugenia uniflora L.). Os extratos produzidos em três temperaturas (30, 45 e 60 °C) foram caracterizados quanto a sua cor instrumental, teor de sólidos solúveis, matéria seca, pH, composição proximal e em relação a sua atividade antioxidante (capacidade redutora, DPPH e ABTS). Os extratos que apresentaram maiores atividades antioxidantes foram utilizados na produção dos filmes. Os filmes foram produzidos pela técnica casting com 8 g de gelatina/100g de solução e glicerol (30 g de glicerol/100 g de gelatina) como plastificante. A quantidade de extrato adicionada foi estudada (0, 50 e 150 g de extrato/100 g de gelatina) para a obtenção das melhores propriedades antioxidantes. Os filmes foram submetidos às seguintes caracterizações: espessura, umidade, solubilidade, cor, opacidade, brilho, permeabilidade ao vapor de água, propriedades mecânicas (tração e perfuração), microestrutura (espectroscopia de força atômica e microscopia eletrônica de varredura), propriedades térmicas (DSC), estrutura química com uso de espectroscopia no infravermelho com transformada de Fourier e determinação da atividade antioxidante (capacidade redutora, DPPH e ABTS). O estudo demonstrou que as amostras vegetais analisadas e as temperaturas de extração escolhidas foram eficientes na produção de extratos com atividade antioxidante, sendo possível sua utilização para produção de filmes ativos. Os extratos que apresentaram maiores atividades antioxidantes foram os extratos de folha de pitangueira, semente de guaraná e hortelã, extraídos a 60 °C, sendo estes escolhidos para a produção dos filmes. Para a caracterização dos filmes, a adição dos extratos em geral não afetou nas propriedades mecânicas (tração e perfuração), umidade e solubilidade, não existindo diferença significativa entre os filmes controles sem extratos e os com extratos. A cor dos filmes sofreu influência na adição dos extratos, sendo que filmes acrescidos com os extratos apresentaram uma tendência mais escura e amarelada que os filmes controles. Entretanto, observou-se que, como esperado, os filmes com extratos demonstraram um aumento na atividade antioxidante e propriedades de barreira, sendo os acrescidos com extrato de folha de pitangueira, os que apresentaram melhores resultados. / Films based on biopolymers containing compounds with antioxidant activity can be used to produce active packaging, which interact with the food, contributing to it conservation and the increasing of the shelf life of the food rich in compounds oxygen sensitive. Plant extracts are rich in polyphenols, known for their antioxidant capacity and are an alternative to the use of synthetic antioxidants. Thus, the general goal of this thesis was to develop gelatin-based films added with vegetal extracts, with physical and functional properties for their use as bioactive packaging. Liquid extracts of Guarana (Paullinia cupana Kunth) seeds, mulberry (Morus nigra L.) leaves carqueja (Baccharis trimera.), peppermint (Mentha piperita) and pitanga (Eugenia uniflora L.) leaves were produced. The extracts produced in three temperatures (30, 45 and 60 °C) were characterized by their antioxidant capacity (total phenols, DPPH and ABTS), instrumental color, soluble solids, dry matter, pH and proximal composition. The extracts with the highest antioxidant activity were used in the film\'s production. The films were produced by the casting technique with 8 g of gelatin / 100 g of solution and glycerol (30 g of glycerol/100 g of gelatin) as plasticizer. The amount of extract added to studied the antioxidant properties were : 0, 50 and 150 g of extract / 100 g of gelatin. The films were subjected to the following characterizations: thickness, mechanical properties (traction and perforation), color, opacity, gloss, humidity, solubility, water vapor permeability, microstructure (Atomic force spectroscopy and scanning electron microscopy), thermal properties (DSC), determination of the antioxidant capacity, determination of reducing capacity and chemical structure using Fourier transform infrared spectroscopy. The study demonstrated that the analyzed plant samples and the chosen extraction temperatures were efficient in the production of extracts with antioxidant activity, being possible its use for the production of active films. Extracts of pitanga leaves, guarana seed and peppermint, produced at 60 °C, were chosen due to their high antioxidant activity. In general, the characterization of the films shown that the addition of the extracts did not affect in the mechanical, solubility and humidity properties, having no significant difference between the control films, with no extract, and those with extracts. The colour of the films were influenced by the addiction of the extract, with the increase of darkness and yellowness of the films, when films were added with the extracts. As expected, it was observed that the films with extracts showed an increase in the antioxidant capacity and barrier properties, being that films added with extracts of pitanga leaves presented the better results.

A thermo-chemical treatment method was used to produce biogenic amorphous silica from South African sugarcane and maize residues. Different fractions of South African sugarcane (leaves, pith, and fiber) were processed for silica production. The biomass samples were leached with either 7 wt% citric acid or 7 wt% sulfuric acid at 353 K for 2 h prior to being rinsed, dried and combusted using a four-step program ranging from room temperature to 873 K in a furnace. The characterization of the pre-treated biomass samples was conducted using thermogravimetric analysis (TG/DTA), X-ray fluorescence analysis (XRF) and elemental analysis (CHN), while the final products were characterized by XRF, X-ray diffraction (XRD), elemental analysis, nitrogen physisorption and scanning electron microscopy (SEM). Citric acid pre-treatment proved to be an attractive alternative to mineral acids. Amorphous biogenic silica was produced from sugarcane leaves in good quality (0.1 wt% residual carbon and up to 99.3 wt% silica content). The produced biogenic silica also had great textural properties such as a surface area of up to 323 m2 g−1, average pore diameter of 5.0 nm, and a pore volume of 0.41 cm3 g−1.

Forward genetic screen was performed in Arabidopsis thaliana to isolate novel genes involved in leaf development. The tarani (tni) mutant was selected for further study based on its unique cup-shaped lamina with +ve Gaussian curvature. We show that the larger size of tni leaves is due to rapid growth rate due to excess and prolonged cell division. We monitored the front of the receding cell division zone as a function of time and showed that the shape of the front is more concave compared to wild type, leading to positive curvature. Application of gibberellic acids (GA) synthesis inhibitor rescued the positive curvature of tni suggesting a role for GA in maintaining leaf flatness. Overexpression of cell cycle inhibitor KRP2 also flattened the leaf, confirming a role of cell division. The floral organs and seed are also larger in the tni mutant. Besides growth, tni trichomes are hyper-branched which usually happens when there is more endoreduplication. We found that the nuclei of tni trichomes are larger than wild type nuclei, suggesting increased DNA content. Genetic interaction studies showed that TNI works independent of other trichome branching genes such as with TRYPTICHON and FURCA1. Map-based cloning showed that tni is positioned on left arm of the 3rd chromosome. Using molecular markers, we narrowed down to interval to a 65 kb region, which codes for 19 genes. Sequencing several of them revealed a G→A transition at the 3rd intron - 4th exon junction of At3g20630 gene. RT-PCR analysis showed the presence of an additional full-length transcript with extra un-spliced 3rd intron. Overexpression of this un-spliced variant in wild type plants produced phenotypes like hyperbranched trichomes and cup-shaped leaves; plus additional phenotypes like organ fusion and organ polarity defects. Complementation and allelic tests confirmed that TNI codes for AtUBP14, an ubiquitin protease. The tni plants have longer stem and roots which grow at faster rate compared to wild type. Confocal microscopic analysis of mature embryos showed that both shoot (SAM) and root apical meristems (RAM) of tni plants are larger in size. In RAM, the numbers of quiescent center (QC) cells and stem cells have increased in tni plants. The tni inflorescence and flowers are bigger than wild type in size. Also the degree of axillary shoots has increased in the tni plants. Overexpression of the splice variant of TNI produced undifferentiated callus-like structures in the shoot apex and in hypocotyl. All these phenotypes show that TNI is involved in meristem proliferation. The tni siliques produced many un-fertilized ovules and shrunken and malformed seeds suggesting gametic and/or embryo lethality. We observed that tni embryos were mis-patterned at various stages of development. Following the cell division pattern shows that cells arising from the ‘basal cell’ of the embryo take apical cell fate in tni embryos. The topmost cell of the suspensor, which is also the precursor cell of RAM, is not specified as hypophysial cell in several tni embryos. In the forward genetic screen, we isolated another mutant called tooth (tth), which has deeper serrations at the leaf margin and narrower leaves compared to wild type. It has been mapped to the longer arm of the 2nd chromosome. Genetic interaction studies show that tth is not allelic to other serration mutants such as serrate and mir164a.

As matas riparias tem efeitos positivos sobre a qualidade da água em rios de baixa ordem. O cultivo e a substituição de plantas de espécies nativas por espécies exóticas, tem efeitos e impactos significativos na biodiversidade de ambientes aquáticos e terrestre. O cultivo de Eucalyptus ocupam áreas de importância biológica e ambiental, causando impactos que ainda são pouco estudados, interferindo no funcionamento dos cursos de água que são dependentes do material foliar disponível nas margens dos rios como fonte de energia e nutrientes. O presente trabalho comparou a decomposição de folhas de espécies nativas (Inga marginata e Matayba elaeagnoides) e folhas de uma espécie exótica (Eucalyptus grandis) durante as estações seca e chuvosa, num riacho localizado no interior da Floresta Nacional de Ipanema, unidade de conservação situada no sudeste do estado de São Paulo. Para a comparação foi utilizado o método de litter bags usando sacos de malha grossa plástica de 10 mm de abertura, por períodos de no máximo 60 dias. Neste período também foi avaliada a colonização de macroinvertebrados bentônicos em cada um dos tratamentos testados. Entre as estações (seca e chuvosa), as perdas de massa foliar foram distintas, assim também existiram diferenças entre as espécies de plantas testadas (Inga marginata, Matayba elaeagnoides e Eucalyptus grandis), sendo sempre maior a perda nas folhas de Inga marginata, seguida das folhas de Eucalyptus grandis e Matayba elaeagnoides, e obtendo os maiores valores de perda de massa na estação chuvosa. Foram identificados 801 indivíduos que colonizaram nos três tratamentos testados na época seca e 556 indivíduos que colonizaram nos tratamentos na época chuvosa, existindo uma relação entre os valores de perda de massa e a abundância de organismos macroinvertebrados para cada um dos tratamentos, sendo os tratamentos com maiores perdas os que tiveram maiores valores de organismos colonizados. Os maiores valores de perda de massa para a folha de Inga marginata foi maior em comparação dos outros tratamentos além de apresentar a maior quantidade de macroinvertebrados aquáticos colonizados nas duas épocas avaliadas. As famílias mais representativas foram: Chironomidae, Hyalellidae, Gomphidae, Calamoceratidae, Leptophlebiidae e Elmidae. Assim os grupos funcionais com maior presença foram os grupos coletor e predador, não existindo diferenças significativas na presença destes grupos. O grupo fragmentador foi o terceiro grupo mais representativo para os três tratamentos e nas duas épocas avaliadas. Nesse tipo de ambiente tropical, a comunidade de fungos e bactérias é rico e diverso, fazendo esse processo biológico, importante para a transformação das folhas em um recurso de maior facilidade de assimilação para seu aproveitamento, tornando assim as folhas mais acessíveis para os organismos invertebrados. Deste modo, por exemplo transformando as folhas de eucalipto em um recurso, sendo estas folhas também acondicionadas para o consumo dos organismos e podendo-se constituir como um recurso adicional para a biota e os organismos se adaptando para a assimilação deste recurso. A presença de folhas de eucalipto pode influenciar na colonização dos organismos invertebrados, podendo interferir no processo natural da colonização de macroinvertebrados aquáticos e alterando a disponibilidade de nutrientes dentro das teias tróficas destes sistemas aquáticos. / The crumbled vegetation has positive effects on water quality in a low river. The crop plants and replacement of native species by exotic species, has significant effects and impacts on biodiversity of aquatic and terrestrial environments. Eucalyptus cultivation occupies areas of biological and environmental importance, causing impacts that are still poorly studied, interfering with the operation of the waterways that depend on available vegetable waste on the banks of rivers as a source of energy and nutrients. This study compared the decomposition of leaves of native species (Inga marginata and Matayba elaeagnoides) and leaves of an exotic species (Eucalyptus grandis) during the dry and rainy seasons in a river located within of the Floresta Nacional de Ipanema, conservation unit located in the southeastern state of São Paulo. For the comparison, the litter bags method was used. Thus, networks of plastic coarse mesh with an opening of 10 mm maximum were used for periods of 60 days. In this period the colonization of bentic macroinvertebrates in each of the treatments was also evaluated. Between the seasons (dry and rainy), losses of plant mass were different among different species of evaluated plants (Inga marginata, Matayba elaeagnoides and Eucalyptus grandis), considering that the loss of leaves of Inga marginata was higher, followed by the Eucalyptus grandis and Matayba elaeagnoides leaves. As a result, the loss of mass in the rainy season was evident. Thus, 801 individuals that colonized the three treatments evaluated in the dry season and 556 individuals that colonized treatments in the rainy season were identified. As consequence, there is a relationship between the values of loss of plant material and abundance of macroinvertebrates organisms for each of the treatments. The treatments with higher losses those who had higher values of colonized organisms. The largest mass loss values for Inga marginata sheet was higher in comparison of other treatments as well as presenting the greatest amount of aquatic macroinvertebrates colonized in the two seasons evaluated. The most representative families were: Chironomidae, Hyalellidae, Gomphidae, Calamoceratidae, Leptophlebiidae and Elmidae. Thus, the functional groups with the largest presence were collectors and predators groups, with no significant differences in the presence of these groups, the shredder group was the third most representative group for the three treatments and two periods evaluated. In this type of tropical environment, the community of fungi and bacteria is abundant and diverse, making of this biological process, an important aspect for the transformation of leaves in a resource ease of assimilation for its use, allowing these to become more accessible for invertebrate organisms. Considering the previous information, this allowed the Eucalyptus leaves transform in a resource available for consumption of organisms and being able to be as an additional resource for biota and organisms adapting to the assimilation of this resource. Thus, the presence of Eucalyptus leaves can influence the colonization of invertebrate organisms, it may interfere with the natural process of colonization of aquatic macroinvertebrates and altering the availability of nutrients in the food chains of these aquatic ecosystems.

O número das espécies microbianas presentes na forragem no ato da colheita é responsável pelo padrão de fermentação da silagem, sendo que o número de unidades formadoras de colônia (ufc) é alterado ao longo do ciclo da cultura e das condições ambientais, e esse fato, influencia fortemente a eficácia de aditivos utilizados no processo. Neste contexto, três experimentos foram conduzidos a fim de caracterizar a população microbiana em plantas de cana-de-açúcar. No primeiro experimento foi caracterizada a população microbiana em plantas de cana-de-açúcar por meio da técnica de plaqueamento. Os tratamentos impostos foram compostos por duas fontes de adubação (NPK e NPK associado a esteco bovino curtido 120kg de N/ha) da cana-de-açúcar colhidas manualmente com vista à quantificação do número de bactérias ácido láticas, leveduras e mofos, bem como relacionar suas freqüências de ocorrência com efeitos ambientais, fonte de fertilizante e estádio de maturação, com colheitas realizadas aos 10, 12, 14 e 18 meses, correspondendo aos meses de Março, Maio, Julho e Novembro de 2009, respectivamente. No segundo experimento foram avaliadas as variáveis biométricas, morfológicas, dinâmica de acúmulo de MS e dos nutrientes em cana-de-açúcar submetida às fontes de fertilização, descritas anteriormente. No terceiro experimento foi realizada avaliação química e bromatológica de cana-de-açúcar submetida à fontes de adubação. O experimento composto por blocos inteiramente casualizados sendo 6 blocos subdivididos em 2 parcelas. Os dados gerados foram analisados pelo procedimento Proc Mixed e Proc NLIN, do programa SAS. Não se observou efeito do tratamento sobre nunhuma variável avaliada. A produtividade média de massa verde variou entre 162 e 188 tMV/ha, podendo ser considerada elevada. O número de folhas verdes partiu de 9,5 aos 10 meses e atingiu cerca de 10 aos 18, o que é esperado, uma vez que ao passar do tempo há aumento da biomassa das plantas. O mesmo se observo para folhas secas, partindo de cerca de zero folhas por planta até cerca de 3 folhas. O peso das folhas também foi crescente. O peso e comprimento do colmo aumentaram de Março para Maio, mas apartir de Maio e Julho esse crescimento foi estagnado, não aumentando significativamente devido à seca. Porém, apartir de Julho esse crescimento voltou a ocorrer com o aumento das chuvas. A cana-de-açúcar apresentou 10 oBrix aos 10 meses, 17oBrix aos 12 meses e 20obrix aos 20 meses. O índice de maturidade aumentou, saindo de 33,46 aos 10 meses e atingindo 88,5 aos 18 meses. Aos 12 meses a cana-de-açúcar apresentou teor de matéria seca de 24%, e aos 18 meses 28%. Os teores de FDN e FDA da planta inteira dimunuiu ao longo do tempo, dos 10 meses (61% e 38,4% de FDN e FDA, respectivamente) para os 12 meses e permanecendo constante até os 18 meses (55% e 35,6% de FDN e FDA, respctivamente). Foi observado que existe correlação positiva entre DIVMS e do oBrix,e a equação, DIVMS = 41,35 + oBrix ; com R2=0,73 e P<0,01 se estabelecendo como ferramenta importante para se estimar a DIVMS. Ao longo do tempo, a contagem de bactérias ácido láticas e de leveduras foi crescente. De forma geral, todas as frações da planta apresentaram contagem numericamente semelhante, partindo de cerca de 4log ufc/gMV aos 10 meses e atingindo cerca de 5,5 log ufc/gMV aos 18 meses. Aos 10 meses a planta inteira de cana-de-açúcar apresentou contagem de leveduras de 4 log ufc/g MV permanecendo constante até os 18 meses, quando atingiu 5,7 log ufc/g MV. A fonte de adubação, quer seja química ou orgânica, não interfere na população de microrganismos, que entretanto, varia em função do período experimental e das condições climáticas. / The numbe of microbial species in forages during the ensiling is responsible for the silage fermentation, and the numbeof colony forming units (cfu) changes during the crop cycle and environmental conditions. It suggests that the initial microbial profile strongly influences the effectiveness of additives used in the process. In this context, we propose three trials to characterize the microbial population in plants of sugarcane. The first trial evaluated the microbial population in plants of sugarcane by the technique of pour plating. The treatment consisted of two levels of fertilization (NPK and NPK associated with solid manure - 120kg N / ha) applied immediately after sugarcane was harvested by hand. The objective was quantify the number of lactic acid bacteria, yeasts and molds, as well as relating their frequencies with environmental effects, fertilization and stage of maturation, with samples taken at 10, 12, 14 and 18 months, corresponding to the months of March, May, July and November 2009. The second trial measured the biometric variables, morphological, the accumulation of dry matter and nutrients in sugarcane subjected the sources of fertilization, as mensioned. The third trial performed chemical assessments of sugarcane subjected to the sources of fertilization. The trials consisted of a completely randomized design with six blocks sub-divided into two plots. Data were analyzed by the procedure Proc Mixed and Proc NLIN of SAS program. There was no effect of treatment in any trial. The fresh yield ranged from 162 to 188tGM/ha which can be considered high. The number of green leaves was increased from 9.5 to 10 to 10 to 18, which is expected, since there is an increase of plant biomass across the time. The same was observed for dead leaves, from about 0 leaves per plant up to 3 leaves. The weight of the leaves was also increased. The weight and length of the stem increased from March to May, and became more intensive from July with increasing rainfall. However, during the dry season (May - July) there was a decreased growing rate.The sugarcane oBrix was 10 at 10 months, 17 oBrix at 12 months and 20oBrix to 20 months. The maturity index increased, from 33.46 to 10 months reaching 88.5 at 18 months. At 12 months the sugarcane showed dry matter content of 24% and 28% at 18 month. The NDF and ADF of the whole plant decreased over time from 10 months (61% and 38.4% NDF and ADF, respectively) for 12 months and remained constant until 18 months (55% and 35 6% NDF and ADF, respectively). A is positive correlation between IVDMD and oBrix was observed, and the equation, IVDMD = 41.35 + oBrix, with R2 = 0.73 and P <0.01 is an important tool to estimate IVDMD. Over time, the counts of lactic acid bacteria were increased. In general, all plant fractions showed similar counts, starting from 4 log cfu/gGM to 10 months and reaching about 5.5 log cfu/gGM to 18 months. At 10 months the whole plant sugarcane had yeast count of 4 log cfu/gGM remaining constant until 18 months, when it reached 5.7 log cfu / g MV. The source of fertilizer, whether chemical or organic, does not interfere in the population of microorganisms, however, varies depending on the experimental period and climatic conditions.