Frutose e Ácido Glicônico são produtos importados empregados em diferentes setores nas áreas química, farmacêutica e alimentícia, representando um mercado de dois milhões de dólares (US$ 2,0 milhões) por ano. Por sua vez, a sacarose pode ser empregada como matéria-prima para a obtenção destes produtos através de conversão enzimátiva empregando invertase e glicose-oxidase. O uso de biorreatores com membrana (MBR) mostra-se interessante em processos enzimáticos, pois, ao serem empregados em processos contínuos permitem, simultaneamente, produção e separação dos produtos, reduzindo a formação de subprodutos e, eventual, inibição da enzima por excesso de substrato ou produtos. A sacarose é convertida em xarope de açúcar invertido (solução equimolar de frutose e glicose) pela invertase (Bioinvert®, enzima comercial), seguido pela oxidação da glicose em ácido glicônico pela ação da glicose oxidase (GO). O processo de conversão multi-enzimático da sacarose foi obtido através da alimentação de sacarose (50 mM) em reator com membrana (MBR) contendo invertase (24 U/mL), glicose-oxidase (0,5 U/mL) e catalase (470 U/mL) e operando com vazão específica de 6,0 h-1, 35ºC e pH 5,5. As condições operacionais otimizadas possibilitaram a conversão completa da sacarose (X = 100 %) e da glicose resultante (Y = 100%) com velocidades específicas de reação de 4,2 mmol/U.h, 0,60 mmol/U.h e 0,00062 mmol/U.h, respectivamente, para a invertase, glicose oxidase e catalase. A respeito da oxidação da glicose, a adição de catalase no meio reacional se fez necessária para minimizar os efeitos inibitórios sobre a GO através do peróxido de hidrogênio formado. / The fructose and gluconic acid are products of great application in chemical, pharmaceutical and food industry. The actual Brazilian market for these compounds is about US$ 2 millions, here as the sucrose, the raw-material used for their production, represents about 2.4% of the Brazil\'s GNP. This conversion increases the value added to the sugarcane, usually marketed as a commodity, because the fructose and gluconic acid are more valuable products than sucrose. The use of membrane bioreactor (MBR), which operates under mild conditions regarding internal pressure, temperature and pH, has been growing along the years for enzyme catalyzed processes. Moreover, in the MBR the reaction and separation of the products occur simultaneously, avoiding the formation of by-products and the eventual inhibition of the enzyme caused by excess of substrate or products. The sucrose is converted to the inverted syrup (an equimolar solution of fructose and glucose) by invertase (in this work was employed Bioinvert®, a commercial invertase) followed by the oxidation of glucose in gluconic acid by the glucose oxidase (GO). The multi-enzymatic conversion of sucrose was attained when carried out under initial substrate of 50mM and invertase, glucose oxidase and catalase concentrations, respectively, of 24.0 U/mL, 0.5 U/mL and 470 U/mL in a membrane reactor utilizing a dilution rate of 6.0 h-1, 35ºC and pH 5.5. The optimized operational conditions led to a conversion yield of 100% for sucrose hydrolysis and glucose oxidation steps resulting in enzyme productivity of 4.2 mmol/U.h, 0.60 mmol/U.h and 0.00062 mmol/U.h, respectively, to invertase, glucose oxidase and catalase. In regard to the glucose oxidation, the addition of catalase in the reaction medium was necessary, in order to minimize the inhibition of the GO by the hydrogen peroxide formed.

A invertase comercial (Bioinvert®) foi imobilizada por adsorção em resinas aniônicas do tipo Dowex® [1x8:50-400, 1x4:50-400 e 1x2:100-400, todos copolímeros estireno-divinilbenzênicos, porém de granulometria (50-400 mesh) e quantidades de ligações cruzadas diferentes (2-8%)] em meio aquoso. A melhor percentagem de adsorção da invertase nas resinas foi observada em pH 5,5 a 32°C, tendo o complexo Dowex®1x4-200/lnvertase apresentado índice de adsorção e coeficiente de imobilização iguais a 100%. Os parâmetros cinéticos e termodinâmicos foram determinados para a invertase solúvel e insolúvel de Bioinvert® e também para a invertase purificada (Fluka®). O complexo Dowex®1 x4-200/Bioinvert® apresentou-se estável durante as reações sem desprendimento da enzima do suporte. Os parâmetros termodinâmicos da forma solúvel e insolúvel da Fluka® foram idênticos aos do Bioinvert®, no entanto, após a imobilização apresentou uma redução de 28% na sua atividade. O estudo da atividade transferásica de ambas as formas de Bioinvert® em diferentes concentrações de sacarose foram analisadas através da cromatografia de camada delgada. A estabilidade operacional e de estocagem foi também determinada para o complexo Dowex®1x4-200/Bioinvert®. / The invertase (trademarked as Bioinvert®) solubilized in deionized water was immobilized by adsorption on anion exchange resins, collectively named Dowex®, [1x8:50-400, 1x4:50-400 and 1x2:100-400, styrene-divinylbenzene copolymers, with different granulometry (50-400mesh) and different degrees of cross-linking (2-8%)]. The best percentage of adsorption of invertase on resins was observed in pH 5.5 at 32°C and the complex Dowex®1x4-200/invertase has shown a coupling yield and an immobilization coefficient equal to 100%. The thermodynamic and kinetic parameters for sucrosehydrolysis for both soluble and insoluble enzyme were evaluated to Bioinvert® and purified invertase purchased from Fluka®. The complex Dowex®/Bioinvert® was stable without any desorption of enzyme from the support during the reaction and having the thermodynamic parameters equal to the soluble formo However, the loss of activity for immobilized Fluka® was found to be 28% when compared to the soluble one. The transfructosylating activity of Bioinvert® in both forms in different concentrations of sucrose was investigated through TLC. In regard to insoluble Bioinvert® its storage and operational stability were also determined.

A conversão enzimática da sacarose pela ação sucessiva da invertase e da glicose oxidase (GOD), permite obter produtos de maior valor agregado, a saber, frutose e o ácido glicônico, dois produtos de amplo uso na indústria farmacêutica, alimentícia e química. Foi estudada a aplicação da invertase imobilizada em resinas aniônicas do tipo Dowex® (um copolímero de poliestireno-divinilbenzeno) sobre a hidrólise da sacarose bem como a oxidação da glicose pela glicose oxidase solúvel ou imobilizada no mesmo suporte em separado (sistema bifásico), utilizando-se um reator de membrana acoplado à membrana de ultrafiltração (100kDa) ou de microfiltração (5µm). Posteriormente, avaliou-se o desempenho de ambas as formas de enzimas, solúveis ou imobilizadas num sistema monofásico empregando o mesmo reator. A bioconversão executada em sistema bifásico permitiu a obtenção de xarope de frutose da ordem de 70% através da separação de glicose e frutose utilizando-se a resina catiônica 50W:8-100. O rendimento de 96,6% e 67,4% para as formas solúveis e imobilizadas respectivamente foram obtidas em sistema monofásico. O não desprendimento das enzimas dos suportes viabilizou o uso da membrana de microfiltração, trazendo vantagens à operação de biorreator com membrana. / The enzymatic conversion of sucrose through a successive action of invertase and glucose oxidase (GOO) allows the obtainment of products with higher commercial value, fructose and gluconic acid, which are widely used in pharmaceutical, food and chemical industries. Invertase and GOO immobilized on Dowex® anionic resin (a polystyrene divinylbenzene copolymer) as well as soluble GOD were used in a membrane bioreactor (MS) for sucrose hydrolysis and glucose oxidation. The MB was coupled with a UF-membrane (100kDa) or a MF-membrane (5µm). The bioconversion was conducted in two steps (biphasic system) as well as in one step (monophasic system). The bioconversion operated in a biphasic system permitted obtaining a fructose syrup with a concentration of about 70% through a separation of glucose and fructose using a cationic resin, 50W:8-100. As for the monophasic system, the yield of 96.6% and 67.4% for soluble and immobilized forms were attained respectively. No leakage of the enzymes from the support allowed the use of a microfiltration membrane, adding advantages to the membrane bioreactor operation.

Made available in DSpace on 2017-07-10T18:08:11Z (GMT). No. of bitstreams: 1 Lexandra Novaki.pdf: 494663 bytes, checksum: dd065fb2df65e19ff5515118352e33a1 (MD5) Previous issue date: 2009-03-16 / The invertase, enzyme found in yeast, above all in the Sacharomyces cerevisiae species, invertebrate, vertebrate, green alga, bacteries, vegetal and fungus, is one of the principal accountable for sucrose hydrolysis to form the inverted syrup, is used by the food industry and pharmacy, in the hydrolyses of rafinose, inulin and gentianose. In the present work, the invertase was produced by Aspergillus casiellus through solid state fermentation, using the industrial waste of soy bran, during a period of 72 h at 25ºC. The enzyme extraction was done by shaking at 120 rpm to period of 1h. The extract obtained was filtered and stocked in freezer. The enzyme recovery was done throughout the protein precipitation with organic solvents, ammonium sulphate and thermal shock. The invertase purification was carried-out in anion-exchange chromatography DEAE-cellulose. It was characterized the enzyme activity of fermented material, the stability relationship to temperature and pH, optimum temperature and pH, effect of the presence of some ions, chelating, redutors agents and detergents, beyond the neutral carbohydrate content determination and specificity to the substrate. The specific enzyme activity was 1029,75 U/mg after the period of 72h of fermentation using soybean bran as substrate. The precipitation with ammonium sulphate and solvents was less efficient than thermal shock for the invertase recovery. The enzyme was purified through thermal shock and anion-exchange chromatography in DEAE-cellulose with a purification factor of 30,30 fold and a yield of 42,70%. The electrophoretic analyses demonstrated that the studied invertase presents molar mass around 25.5 kDa. After submitted to the purification steps, the enzyme showed optimum pH 4.0 and temperature 70ºC. In relation to the stability, after 24 hours of incubation at pH 4.0, the invertase maintained 50% of residual activity. The invertase showed thermalstability at 60 70ºC not arrive the half-life time in 240 minutes was observed 40% of the catalytic capacity remained. In temperatures of 75-85ºC, 40% of residual activity were maintained after 4h of incubation. The partially purified invertase didn´t need metallic ions for catalytic activity, but the presence of Al3+ and Cu+ improved the activity, and Sn2+ challenged decrease in the activity. Sugar content corresponded to 36.57% of total mass protein. The invertase of A. casiellus demonstrated specificity to sucrose. For purification of the invertase of Aspergillus casiellus, induced through soybean bran, were needed 2 steps made up anion-exchange chromatography DEAE-cellulose, which turn the process practible and with few time despended relationship to the others process. The use of agroindustrial waste to obtain invertase is becoming an instrument to development of clean technology, converting the waste in a valuable product. / A invertase, enzima encontrada em leveduras, sobretudo na espécie Sacharomyces cerevisiae, invertebrados, vertebrados, algas verdes, bactérias, vegetais e fungos, é uma das principais responsáveis pela hidrólise da sacarose para a formação de açúcar invertido e é utilizada por indústrias alimentícias e farmacêuticas, na hidrólise da rafinose, inulina e gentianose. No presente trabalho objetivou-se a purificação e caracterização parcial da invertase, obtida por fermentação em estado sólido. A invertase foi produzida por Aspergillus casiellus utilizando como substrato farelo de soja por um período de 72 a 220h, a 25 ºC. A extração das enzimas foi feita utilizando shaker a 120 rpm por um período de 1h. O extrato obtido foi filtrado e estocado em freezer. A recuperação da enzima invertase foi feita através de precipitação das proteínas com solventes orgânicos, sulfato de amônio e através de choque térmico. A purificação da invertase foi conduzida em coluna de DEAE-celulose. Foram caracterizados atividade enzimática do material fermentado, a estabilidade quanto à temperatura e ao pH, temperatura e pH ótimos, efeito da presença de íons, quelantes, agentes redutores e detergentes, além da determinação de carboidratos neutros e especificidade ao substrato. A atividade específica do extrato bruto foi de 1029,75 após um período de 72h de fermentação usando o farelo de soja como substrato. A recuperação por precipitação por sulfato de amônio e solventes foi menos eficiente que um choque térmico para a recuperação da invertase do extrato bruto. A enzima foi parcialmente purificada através de precipitação por choque térmico (utilizando sobrenadante) e cromatografia de troca aniônica em DEAE-celulose com um fator de purificação de 30,30 vezes e rendimento de 42,70%. As análises eletroforéticas estimaram a massa molecular da invertase estudada em torno de 25,5 kDa. Após ser submetida às etapas de purificação, a enzima manteve as condições ótimas de pH 4,0 e de temperatura 70 ºC. Após 24 horas de incubação em pH 4,0, a invertase homogênea manteve 50% de atividade enzimática residual. A invertase demonstrou termoresistência em temperaturas de 60 a 70ºC não atingindo o tempo de meia vida em 240 minutos. Nas temperaturas de 75 a 80ºC foram mantidas aproximadamente 40% da atividade residual após 4 horas de incubação. A invertase parcialmente purificada não demonstrou a necessidade de íons metálicos para a ativação catalítica, mas a presença de íons Al3+ e Cu+ provocaram aumento da atividade, enquanto que Sn2+ diminuiu a atividade catalítica. O conteúdo de carboidratos corresponde a 36,57% da massa total de proteína. A invertase do A. casiellus demonstrou ser especificidade à sacarose. Para a purificação da invertase de Aspergillus casiellus, induzido por farelo de soja, foram necessários 2 passos constituídos de colunas cromatográficas de troca-iônica DEAE-Celulose, o que torna o processo viável e com tempo despendido em relação a outros processos. O uso de resíduo agroindustrial para obtenção da invertase torna-se uma ferramenta para o desenvolvimento de tecnologias limpas, convertendo o resíduo em questão em um produto de maior valor agregado.

Le mildiou est une maladie de la vigne affectant les feuilles et les baies et pouvant entraîner des diminutions importantes de la quantité et de la qualité de la vendange. L'agent responsable, Plasmopara viticola, un oomycète biotrophe obligatoire, provoque différentes altérations physiologiques au niveau de la feuille infectée. D'une part, il y a des accumulations anormales d'amidon au niveau des taches d'huile, symptômes caractéristiques de la maladie. D'autre part, l'infection induit une dérégulation des mouvements stomatiques. Les stomates, ouvertures naturelles permettant les échanges gazeux entre la plante et l'atmosphère, restent anormalement ouverts de nuit et ne se ferment plus en réponse à un stress hydrique ou à un traitement à l'ABA. Les deux parties de cette thèse avaient pour objectif de i) expliquer l'origine et le mécanisme de l'accumulation d'amidon et ii) isoler et identifier le facteur responsable de la dérégulation stomatique. Les modifications du métabolisme de l'amidon au niveau des feuilles infectées ont d'abord été étudiées à l'aide d'une analyse transcriptomique afin d'identifier les gènes codant pour des enzymes impliquées dans ce métabolisme dont l'expression était affectée. Après validation de deux gènes de référence, une étude par qRT-PCR a permis de vérifier les changements d'expression de certains de ces gènes au cours de la cinétique de l'infection, puis les activités des enzymes correspondantes ont été déterminées. Ces approches, complétées par des dosages de l'amidon et des sucres solubles ainsi que la mesure de l'activité photosynthétique, ont permis de corréler différents événements avec l'infection. D'après nos résultats, l'accumulation de l'amidon dans les taches d'huile pourrait résulter d'une augmentation de sa synthèse liée à l'augmentation de l'activité AGPase, combinée à une diminution de sa dégradation par modification de l'activité des amylases. Une diminution de la photosynthèse et une augmentation de l'activité invertase traduisent une transition source-puits au niveau des taches d'huile. La deuxième partie de la thèse a porté sur la recherche d'une molécule soluble d'origine végétale ou pathogène, secrétée dans l'apoplasme au cours de l'infection et responsable du dysfonctionnement stomatique. Sur " épidermes isolés ", les fluides apoplastiques extraits de feuilles infectées induisent une ouverture stomatique à l'obscurité et contrecarrent la fermeture induite par l'ABA à la lumière, mimant ainsi la dérégulation observée sur feuille entière. Le composé actif serait une protéine assez stable, de taille >50 kDa et modifiée post-traductionellement par une glycosylation essentielle à son activité sur l'ouverture stomatique. Après séparation par chromatographie d'exclusion des fluides apoplastiques de feuilles infectées, deux fractions ont été retenues : une présentant une forte activité sur l'ouverture stomatique, et une seconde sans activité. Neuf protéines de vigne ont été identi fiées spécifiquement dans la fraction active après une analyse comparative de ces deux échantillons par spectrométrie de masse. Des expérimentations supplémentaires seront nécessaires pour purifier et identifier la protéine recherchée.

ABSTRACT IMMOBILIZATION OF INVERTASE, POLYPHENOL OXIDASE AND GLUCOSE OXIDASE IN CONDUCTING COPOLYMERS OF THIOPHENE-CAPPED POLYTETRAHYDROFURAN AND PYRROLE B&ouml / y&uuml / kbayram, AySe Elif Ph.D., Department of Chemistry Supervisor: Prof. Dr. Levent Toppare January 2005, 123 pages Immobilization of invertase, polyphenol oxidase (PPO) and glucose oxidase (GOD) enzymes were performed in electrochemically synthesized two types of conducting copolymers. One end and two end thiophene-capped polytetrahydrofuran (TPTHF-1 and TPTHF-2) were copolymerized with pyrrole under conditions of constant potential electrolysis. The copolymers were characterized by thermal, spectroscopic and scanning electron microscopy analyses. Immobilization was carried out via entrapment of enzymes in two types of matrices during the copolymerization of pyrrole with the insulating polymers in the presence of sodium dodecyl sulphate (SDS). Kinetic parameters: Maximum reaction rate (Vmax) and Michaelis-Menten constant (Km) were determined for the enzyme electrodes. Temperature optimization, pH optimization, operational stability and shelf-life of the enzyme electrodes were investigated. Enzyme electrodes of polyphenol oxidase and glucose oxidase were used to determine the amount of their substrates in samples. Polyphenol oxidase converts mono and diphenols to quinone. Amount of phenolic compounds in two kinds of wines were determined by analyzing the quinone amount. Glucose oxidase converts &amp / #61538 / -D-glucose to D-glucono-1,5-lactone. Glucose amount was determined in two kind of factory-produced orange juices by analyzing D-glucono-1,5-lactone.

Made available in DSpace on 2014-06-11T19:32:58Z (GMT). No. of bitstreams: 0 Previous issue date: 2005-08Bitstream added on 2014-06-13T19:22:54Z : No. of bitstreams: 1 arauco_emr_dr_botfmvz.pdf: 810943 bytes, checksum: 6acc5913059689bd4615fef648271508 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Universidade Estadual Paulista (UNESP) / O néctar é coletado pelas operárias coletoras e transportado em sua vesícula nectarífera para a colméia. Durante a sua transformação em mel, o néctar recebe enzimas provenientes das glândulas hipofaringeanas, as quais atuam em seu processamento. A invertase é uma das enzimas adicionadas ao néctar e promove a inversão da sacarose em frutose e glicose. Com o objetivo de se conhecer o pH ideal dessa enzima nas abelhas operárias foram extraídas as glândulas hipofaringeanas de abelhas com oito dias de idade. O método utilizado para a dosagem da invertase foi o do ácido 3-5þ dinitrosalicílico (DNS) para determinação de açúcares redutores, utilizando-se medida de leitura espectrofotométrica em absorbância da glicose em comprimento de onda (?) igual a 540nm. De acordo com os resultados, verificou-se que a atividade catalítica da enzima foi mais eficiente no pH 5,0 e em 40 minutos de reação em comparação com outros valores de pH e tempos de reação. Não foi possível a reutilização das glândulas. Desta forma, concluiu-se que a enzima invertase aumentou sua eficiência em função do tempo de reação de forma linear e significativa em pH 5,0. / The nectar is collected by the collecting workers that carry it in its nectar gallbladder to the beehive. During its transformation in honey, the nectar receives enzymes proceeding from the hypopharyngeal glands, which act in its processing. Invertase is one of enzymes added to the nectar and promotes the inversion of sacarose in fructose and glucose. With the aim of knowing the ideal pH, in the worker bees, the bee's hypopharyngeal glands with eight days of age had been extracted. The used method, for the dosage of invertase, had been the 3-5 ' dinitrosalisilic acid for determination of reducing sugars, using absorbance spectrophotometry data from the glucose in ? equal to 540nm. According to the results, it had been verified that the enzyme catalytic activity had been more efficient in pH 5,0 and in 40 minutes of reaction, comparing to other pH values and times of reaction. The glands reuse had not been possible. At this way, it was concluded that the invertase enzyme had increased its efficiency by the reaction time at linear and significant way in pH 5,0.

Resumo: O néctar é coletado pelas operárias coletoras e transportado em sua vesícula nectarífera para a colméia. Durante a sua transformação em mel, o néctar recebe enzimas provenientes das glândulas hipofaringeanas, as quais atuam em seu processamento. A invertase é uma das enzimas adicionadas ao néctar e promove a inversão da sacarose em frutose e glicose. Com o objetivo de se conhecer o pH ideal dessa enzima nas abelhas operárias foram extraídas as glândulas hipofaringeanas de abelhas com oito dias de idade. O método utilizado para a dosagem da invertase foi o do ácido 3-5þ dinitrosalicílico (DNS) para determinação de açúcares redutores, utilizando-se medida de leitura espectrofotométrica em absorbância da glicose em comprimento de onda (?) igual a 540nm. De acordo com os resultados, verificou-se que a atividade catalítica da enzima foi mais eficiente no pH 5,0 e em 40 minutos de reação em comparação com outros valores de pH e tempos de reação. Não foi possível a reutilização das glândulas. Desta forma, concluiu-se que a enzima invertase aumentou sua eficiência em função do tempo de reação de forma linear e significativa em pH 5,0. / Abstract: The nectar is collected by the collecting workers that carry it in its nectar gallbladder to the beehive. During its transformation in honey, the nectar receives enzymes proceeding from the hypopharyngeal glands, which act in its processing. Invertase is one of enzymes added to the nectar and promotes the inversion of sacarose in fructose and glucose. With the aim of knowing the ideal pH, in the worker bees, the bee's hypopharyngeal glands with eight days of age had been extracted. The used method, for the dosage of invertase, had been the 3-5 ' dinitrosalisilic acid for determination of reducing sugars, using absorbance spectrophotometry data from the glucose in ? equal to 540nm. According to the results, it had been verified that the enzyme catalytic activity had been more efficient in pH 5,0 and in 40 minutes of reaction, comparing to other pH values and times of reaction. The glands reuse had not been possible. At this way, it was concluded that the invertase enzyme had increased its efficiency by the reaction time at linear and significant way in pH 5,0. / Orientador: Sílvia Regina Cunha Funari / Coorientador: Carlos Ducatti / Doutor

Almost all conifer ovules produce a liquid secretion as part of reproduction. This secretion, termed an ovular secretion, is produced during ovule receptivity and is involved in pollen capture and transport. Historically, examinations of the ovular secretion have focused on how they are part of pollination mechanisms. As a result, the chemical composition of the ovular secretion has not been examined systematically. Investigations into the constituents of the ovular secretion were limited to analyses for simple water soluble compounds such as sugars, minerals, amino acids and organic acids. More recently, the protein component of the secretion has been investigated using mass spectrometry-based proteomics. Proteins involved in processes such as carbohydrate modification, proteolysis, and defence have been identified in conifer ovular secretions. This biochemical complexity suggests a broader view of the function of the ovular secretion is warranted. However, protein identifications only provide putative information on function. Functional characterization of these proteins is needed in order to fully understand how they contribute to ovular secretion function. The research outlined in this dissertation describes the first functional characterizations of proteins found in conifer ovular secretions. Three proteins - invertase, chitinase, and thaumatin-like protein - were characterized in the ovular secretions of Douglas-fir (Pseudotsuga menziesii) and hybrid yew (Taxus × media). The Douglas-fir ovular secretion is capable of converting sucrose to glucose and fructose, confirming that invertases present in the secretion are functional. The invertase activity was maximal at pH 4.0. Activity was 77% of maximal at pH 4.5, the physiological pH. This indicates that post-secretory hydrolysis of sucrose occurs in situ in the Douglas-fir ovular secretion. Invertases in the ovular secretion are likely involved in controlling the movement of carbohydrates to developing pollen and could facilitate pollen selection. Chitinases present in the Douglas-fir ovular secretion are functional at physiological conditions. All three modes of chitinolytic activity, i.e. endochitinase, chitobiosidase and β-N-acetylglucosaminidase, were detected at physiological pH. β-N-acetylglucosaminidase activity was 80 % of maximal at physiological pH. Chitinases are pathogenesis-related proteins capable of hydrolysing chitin in fungal cell walls. These results suggest the ovular secretion is capable of defending the ovule against infection by phytopathogens. Thaumatin-like protein was immunolocalized to the cell wall and amyloplasts in Douglas-fir and yew nucellar tissue in a pattern consistent with a defensive role. It was also localized to the cell wall of fungal spores and germinating hyphae that were present in the micropyle of a yew ovule. These results provide additional evidence for an antifungal role for the ovular secretion. Functioning enzymes involved in pollen-ovule interactions and ovule defence are present in the conifer ovular secretion. The ovular secretion has functions beyond pollen capture. A revised functional model for the conifer ovular secretion is proposed. / Graduate / 2021-08-17

No description available.