Diante do atual cenário de expansão dos plantios geneticamente modificados e da carência de metodologias no Brasil para prever os seus impactos foi desenvolvimento um método intitulado Impactos-PGM para Avaliação de Impactos Ambientais e Alimentares de Plantas Geneticamente Modificadas utilizando como ferramenta um Software de aplicação geral adequado e modificado com esta finalidade. Para tanto, foi inicialmente realizado o levantamento de indicadores de impactos a partir da literatura. Esta consulta foi realizada pela aplicação de questionário de acordo com a técnica delphi. Posteriormente, os especialistas foram entrevistados presencialmente: foram apresentados os indicadores mais representativos para a avaliação dos impactos das PGMs, possibilitando a elaboração do método Impactos-PGM por meio da adequação do Software Impactos. / In face of the current expansion scenario of genetically modified crops and the lack of methodologies in Brazil to predict its impacts, the present work has been proposed to develop a method entitled Impactos-GMP for \"Food and Environmental Assessment of Genetically Modified Plants (GMPs) Impacts using as tool a software of general application suitable and modified for this purpose. Thus, the survey was first conducted by identifying impact indicators from the literature. This survey was held through the application of a questionnaire according to the delphi technique. Subsequently, the researchers were interviewed in person to whom were presented the most representative indicators for GMPs assessing impacts, enabling the development of the method Impactos-GMP with the adequacy of the iImpacts software.

A dificuldade de transformação de tecido adulto de espécies lenhosas é uma barreira ainda a ser contornada para a maioria das cultivares de laranja doce de importância na citricultura brasileira. Esta dificuldade está relacionada ao alto nível de contaminação, ao baixo potencial regenerativo e à baixa capacidade de transformação de tecidos adultos de espécies lenhosas. Assim, o objetivo deste trabalho foi otimizar o protocolo de transformação genética de tecido adulto de três cultivares de laranja doce. Com ajustes no cultivo in vitro, foi possível obterem-se gemas adventícias transgênicas via Agrobacterium tumefaciens de três cultivares de laranja doce (Citrus sinensis L. Osbeck), Hamlin, Pêra e Valência, utilizando material adulto como fonte de explantes. As gemas transgênicas foram identificadas como transgênicas pelo teste histoquímico GUS e confirmados pela análise de PCR, a qual mostrou a amplificação de um fragmento 817 pb, correspondente a parte do gene uidA amplificada. A utilização de meio de co-cultivo com a adição de auxinas e citocininas (2,0 mg L-1 de 2,4 D; 2,0 mg L-1 de AIA e 1,0 mg L-1 de 2-iP), foi o mais eficiente para as três cultivares avaliadas. Meios de seleção com altas concentrações de citocinina (3 mg L-1) favoreceram a regeneração e, conseqüentemente, a transformação de gemas adventícias para as três cultivares estudadas. O uso da sonicação não foi eficiente para diminuir a contaminacão endofítica. Além disso, esta operação diminuiu a eficiência de transformação dos explantes. As eficiências médias de transformação genética encontradas para as três cultivares foram 2,5% para laranja Hamlin, 1,4% para laranja Pêra e 3,7% para laranja Valência. / The difficulty of tissue transformation of adult woody species is still an obstacle to be overcome for most sweet orange cultivars in the Brazilian citrus industry. This difficulty is related to the high level of contamination, low regenerative potential and low transformation capacity of adult tissues of woody species. Thus, the objective of this work was to optimize the protocol for genetic transformation of adult tissue of three cultivars of sweet orange. Therefore, with in vitro culture adjustments, it was possible to obtain transgenic adventitious buds though Agrobacterium tumefaciens of three sweet orange cultivars (Citrus sinensis L. Osbeck), Hamlin, Pêra and Valencia, using adult material as explants source. The transgenic buds were identified as transgenic by the GUS histochemical test and confirmed by the PCR analysis, which showed fragment amplification of 817 bp corresponding to the part of the amplified uidA gene. The use of co-culture media rich in auxins and cytokinins (2.0 mg L-1 of 2.4 D, 2.0 mg L-1 of AIA and 1.0 mg L-1 of 2-iP) was more efficient for all three cultivars. Selection media with high concentrations of cytokinin (3 mg L-1) promoted the regeneration and, consequently, the transformation of adventitious buds in the three cultivars. The use of sonication was not effective to reduce endophytic contamination. Moreover, this procedure reduced transformation efficiency of explants. The average efficiencies of genetic transformation for the three varieties were 2.5% for Hamlin, 1.4% for Pêra and 3.7% for Valência.

A dificuldade de transformação de tecido adulto de espécies lenhosas é uma barreira ainda a ser contornada para a maioria das cultivares de laranja doce de importância na citricultura brasileira. Esta dificuldade está relacionada ao alto nível de contaminação, ao baixo potencial regenerativo e à baixa capacidade de transformação de tecidos adultos de espécies lenhosas. Assim, o objetivo deste trabalho foi otimizar o protocolo de transformação genética de tecido adulto de três cultivares de laranja doce. Com ajustes no cultivo in vitro, foi possível obterem-se gemas adventícias transgênicas via Agrobacterium tumefaciens de três cultivares de laranja doce (Citrus sinensis L. Osbeck), Hamlin, Pêra e Valência, utilizando material adulto como fonte de explantes. As gemas transgênicas foram identificadas como transgênicas pelo teste histoquímico GUS e confirmados pela análise de PCR, a qual mostrou a amplificação de um fragmento 817 pb, correspondente a parte do gene uidA amplificada. A utilização de meio de co-cultivo com a adição de auxinas e citocininas (2,0 mg L-1 de 2,4 D; 2,0 mg L-1 de AIA e 1,0 mg L-1 de 2-iP), foi o mais eficiente para as três cultivares avaliadas. Meios de seleção com altas concentrações de citocinina (3 mg L-1) favoreceram a regeneração e, conseqüentemente, a transformação de gemas adventícias para as três cultivares estudadas. O uso da sonicação não foi eficiente para diminuir a contaminacão endofítica. Além disso, esta operação diminuiu a eficiência de transformação dos explantes. As eficiências médias de transformação genética encontradas para as três cultivares foram 2,5% para laranja Hamlin, 1,4% para laranja Pêra e 3,7% para laranja Valência. / The difficulty of tissue transformation of adult woody species is still an obstacle to be overcome for most sweet orange cultivars in the Brazilian citrus industry. This difficulty is related to the high level of contamination, low regenerative potential and low transformation capacity of adult tissues of woody species. Thus, the objective of this work was to optimize the protocol for genetic transformation of adult tissue of three cultivars of sweet orange. Therefore, with in vitro culture adjustments, it was possible to obtain transgenic adventitious buds though Agrobacterium tumefaciens of three sweet orange cultivars (Citrus sinensis L. Osbeck), Hamlin, Pêra and Valencia, using adult material as explants source. The transgenic buds were identified as transgenic by the GUS histochemical test and confirmed by the PCR analysis, which showed fragment amplification of 817 bp corresponding to the part of the amplified uidA gene. The use of co-culture media rich in auxins and cytokinins (2.0 mg L-1 of 2.4 D, 2.0 mg L-1 of AIA and 1.0 mg L-1 of 2-iP) was more efficient for all three cultivars. Selection media with high concentrations of cytokinin (3 mg L-1) promoted the regeneration and, consequently, the transformation of adventitious buds in the three cultivars. The use of sonication was not effective to reduce endophytic contamination. Moreover, this procedure reduced transformation efficiency of explants. The average efficiencies of genetic transformation for the three varieties were 2.5% for Hamlin, 1.4% for Pêra and 3.7% for Valência.

Abstract not available

Municipal treated wastewater entering rivers contain biologically active pharmaceuticals capable of inducing effects in aquatic life. Phytoremediation of three of these pharmaceuticals and an herbicide was investigated using Sandbar willow (Salix exigua) and Arabidopsis thaliana. Both plants were effective at removing compounds from solution, with removal of 86% of the synthetic estrogen, 17α-ethynylestradiol, 65% of the anti-hypertensive, diltiazem, 60% of the anti-convulsant, diazepam (Valium®), and 51% of the herbicide atrazine, in 24 hours. Distribution of compounds within roots and shoots, in soluble and bound forms, differed among compounds. Uptake and distribution of pharmaceuticals within the study plants confirmed pharmaceutical behaviour can be predicted based on a physiochemical property, their octanol-water partitioning coefficients. An effective method for detection of 17α-ethynylestradiol within surface water using solid phase extraction and gas chromatography-mass spectrometry was developed. Previously unreported breakdown of 17α-ethynylestradiol into another common estrogen, estrone, during preparative steps and gas chromatography was resolved. / xv, 216 leaves ; 29 cm.

O maracujá-doce (Passiflora alata Curtis) é uma espécie nativa no Brasil. Seu cultivo tem crescido nos últimos anos devido a sua valorização no mercado in natura e seus usos na fitoterapia. Entretanto, os cultivos comerciais enfrentam problemas devido a ocorrência da bacteriose (Xanthomonas axonopodis pv. passiflorae). O patógeno é endêmico no país, apresentando considerável variabilidade genética em suas populações naturais. Este trabalho teve como objetivo identificar QTL relacionados à resistência de P. alata à bacteriose em uma população F1 segregante oriunda do cruzamento entre acessos não endogâmicos. Foram avaliados os caracteres: área total da folha (TA), idade de queda da folha inoculada (IK), área total da lesão foliar (LA), área de clorose foliar (CA) e área da necrose foliar (NEA). Apenas um dos isolados apresentou diferenças de severidade em relação aos demais, sendo o menos agressivo (PA8-2). A inoculação do isolado M-129 mostrou que há variação significativa na resposta da população ao patógeno, sendo possível a identificação de genótipos transgressivos. A herdabilidade dos caracteres variou de 45% a 71%%. Foi construído um mapa de ligação integrado com 1.786 cM e uma densidade média de 4,5 cM. A análise de marcas individuais indicou a associação de 51 marcas aos fenótipos avaliados. O mapeamento de QTL, realizado por intervalo composto e utilizando uma estratégia diferenciada para populações F1 segregantes, identificou regiões associadas a 26 QTL para os cinco caracteres avaliados, sendo 16 deles referentes à LA, CA e NEA. A variação fenotípica explicada individualmente pelos marcadores variou de 0,8% a 16,7%. Sugere-se que a resistência à bacteriose é quantitativa, com predominância de efeitos genéticos aditivos. / The sweet passion fruit (Passiflora alata Curtis) is a specie native to Brazil. Its cultivation has increased in recent years due to its market valuation in natura and their uses in herbal medicine. However, crops are facing problems due to the occurrence of bacterial blight (Xanthomonas axonopodis pv. passiflorae). The pathogen is endemic in the country, with considerable genetic variability in their natural populations. This study aimed to identify QTL related to resistance of P. alata to bacterial blight in an F1 segregant population from the cross between outbred accessions. Five traits were evaluated: total area of the leaf (TA), age of inoculated leaf fall (IK), area of the leaf´s lesion (LA), area of the leaf´s chlorosis (CA) and area of the leaf´s necrosis (NEA). Only one of the isolates showed differences in severity in relation to others, being the least aggressive (PA8-2). The inoculation of the isolate M-129 showed significant variation in population response to the pathogen, making it possible to identify transgressive genotypes. The heritability of characters ranged from 45% to 71%. An integrated linkage map was constructed, with a length of 1,786 cM and an average density of 4.5 cM. The analysis of individual marks indicated the association of 51 markers to phenotypes. The QTL mapping was performed using composite interval and a special strategy for segregating F1 populations, identified 26 regions associated with QTL for the five traits, 16 of them related to LA, CA and NEA. The phenotypic variation explained by individual markers ranged from 0,8% to 16,7%. It is suggested that the resistance to bacterial blight is quantitative, with a predominance of additive genetic effects.

A semelhança entre os eventos da embriogênese zigótica e somática permite que sejam estabelecidos parâmetros para o acompanhamento destes dois processos. Neste trabalho foi estudada a embriogênese somática em Ocotea catharinensis, nas fases de maturação e germinação, além do processo germinativo da semente zigótica, avaliando-se parâmetros bioquímicos, e a expressão diferencial de proteínas. O tratamento com ABA + PEG em culturas de embriões somáticos apresentou variações semelhantes a apresentadas por embriões zigóticos no estádio de maturação. Não se observou a germinação de embriões somáticos, contudo verificou-se que a desidratação prévia promoveu importantes alterações bioquímicas. Durante a germinação, não se observou à síntese de novas proteínas no embrião zigótico. Os estudos proteômicos no desenvolvimento da semente permitiram a seleção de polipeptídios, marcadores estádio-específicos. Os resultados obtidos possibilitam a otimização e melhor entendimento das etapas da embriogênese somática, em espécies com sementes recalcitrantes, como O. catharinensis. / The great similarity among the events of zygotic and somatic embryogenesis allows the establishment of parameters for accompaniment of these processes. In the present work it was studied the somatic embryogenesis in Ocotea catharinensis, in the maturation and germination phases, and the zygotic embryogenesis. The biochemical parameters and differential expression of proteins were evaluated: The treatment with ABA + PEG presented similar variations for zygotic embryos in the maturation stage. The germination was not observed for somatic embryos; however, it was verified that the previous dehydration promoted important biochemical alterations. With relation to the zygotic embryo, throughout the germination process, the synthesis of new proteins was not observed. The proteomic studies carried out throughout seed development, allowed the selection of polypeptides with differential expression. The results obtained open perspectives for the methodology optimization of the somatic embryogenesis, for species with recalcitrant seeds, like O. catharinensis.

Diante do atual cenário de expansão dos plantios geneticamente modificados e da carência de metodologias no Brasil para prever os seus impactos foi desenvolvimento um método intitulado Impactos-PGM para Avaliação de Impactos Ambientais e Alimentares de Plantas Geneticamente Modificadas utilizando como ferramenta um Software de aplicação geral adequado e modificado com esta finalidade. Para tanto, foi inicialmente realizado o levantamento de indicadores de impactos a partir da literatura. Esta consulta foi realizada pela aplicação de questionário de acordo com a técnica delphi. Posteriormente, os especialistas foram entrevistados presencialmente: foram apresentados os indicadores mais representativos para a avaliação dos impactos das PGMs, possibilitando a elaboração do método Impactos-PGM por meio da adequação do Software Impactos. / In face of the current expansion scenario of genetically modified crops and the lack of methodologies in Brazil to predict its impacts, the present work has been proposed to develop a method entitled Impactos-GMP for \"Food and Environmental Assessment of Genetically Modified Plants (GMPs) Impacts using as tool a software of general application suitable and modified for this purpose. Thus, the survey was first conducted by identifying impact indicators from the literature. This survey was held through the application of a questionnaire according to the delphi technique. Subsequently, the researchers were interviewed in person to whom were presented the most representative indicators for GMPs assessing impacts, enabling the development of the method Impactos-GMP with the adequacy of the iImpacts software.

Die Pflanzenbiotechnologie ermöglicht die nachhaltige Gewinnung pflanzlicher Wertstoffe mittels innovativer biotechnologischer Methoden. Bisher mangelt es auf diesem Gebiet jedoch an Grundlagenwissen und aussagekräftigen Studien, z. B. zur Anwendung biotechnologischer Standardverfahren beim Respirationsmonitoring. Im Rahmen der vorliegenden Arbeit werden grundlegende Untersuchungen zur Erzeugung (Induktion) pflanzlicher in-vitro-Kulturen und zu geeigneten Kultivierungssystemen sowie Prozessüberwachungsstrategien vorgestellt und diskutiert. Als Modellsystem dient die Einjährige Sonnenblume Helianthus annuus. Die Induktion pflanzlicher Zellkulturen (Kallus und Suspensionen) mit photomixotrophem Stoffwechsel wurde unter unterschiedlichen Bedingungen untersucht und geeignete Induktionsparameter ermittelt. Sowohl pflanzliche Gewebekulturen (Hairy roots) als auch die erzeugten photomixotrophen und heterotrophe Suspensionen konnten in verschiedenen Reaktorsystemen erfolgreich kultiviert und die Produktbildung nachgewiesen werden. Protokolle zu Induktion sowie Erhaltung von Zell- und Gewebekulturen von H. annuus wurden etabliert. Ein modernes Prozessüberwachungssystem für Schüttelkolben, das RAMOS® (Respiration Activity Monitoring System®) wurde erstmals umfassend für Untersuchungen des Wachstumsverhaltens und zum Screening pflanzlicher Zell- und Gewebekulturen eingesetzt. Dabei wurde die Problematik der Verdunstung (Evaporation) aus den Kulturgefäßen als signifikant bei den langen Kultivierungen von pflanzlichen in-vitro-Kulturen diagnostiziert und ein Modell zur Korrektur der Atmungstransferraten entwickelt. Erstmalig in der Pflanzenbiotechnologie kam das RAMOS® für Studien mit Zell- und Gewebekulturen von H. annuus im Speziellen sowie für Untersuchungen von Hairy roots im Allgemeinen zum Einsatz. Mit Hilfe der vorliegenden Arbeit werden relevante Kriterien zur Anwendung des innovativen Messsystems RAMOS® im Rahmen pflanzenbiotechnologischer Untersuchungen vorgestellt. Es wird ein Überblick über geeignete Kultivierungssysteme und zu publizierten Modellierungsstrategien für Applikationen in der Pflanzenbiotechnologie gegeben. Ein Literaturüberblick zu publizierten Modellierungsstrategien mit pflanzenbiotechnologischem Bezug vervollständigt die Arbeit. / Plant biotechnology enables a sustainable production of valuable plant resources using innovative biotechnological methods. However, a comprehensive knowledge base as well as significant studies, e. g. concerning the application of biotechnological standard procedures of respiration monitoring, are missing so far. In this work, basic investigations regarding the induction of plant in vitro cultures and appropriate cultivation systems as well as process monitoring strategies will be introduced and discussed. The annual sunflower Helianthus annuus serves as biological model system. The induction of plant cell cultures (callus and suspension) with photomixotroph metabolism was investigated at different conditions and appropriate induction parameter were determined. Both, plant tissues (Hairy roots) and induced photomixotroph as well as heterotrophic suspensions were cultivated successfully in various reactor systems. The production of desired metabolites was proven. Protocols concerning induction respectively maintenance of cell and tissue cultures of H. annuus have been established. For extensive investigations of growth behavior and for screening of plant cell and tissue cultures, a modern process monitoring tool for shake flasks, the RAMOS® (Respiration Activity Monitoring System®), was used for the first time. Thereby, the problem of evaporation off the culture vessels was identified as significant for time-intensive cultivations of plant in vitro cultures. A model for the correction of respiration transfer rates has been developed. For the first time in plant biotechnology, the RAMOS® has been applied for studies with H. annuus in special, and for studying the growth and respiration behavior of Hairy roots in general. With the help of the present work, relevant criteria concerning the application of the innovative measuring system RAMOS® for plant biotechnological investigations will be given. Furthermore, a survey over appropriate cultivation systems and published modelling strategies in plant biotechnology are introduced. A literature survey concerning model strategies regarding plant biotechnology completes this work.

Plant cell and tissue cultivations are of growing interest for the production of structurally complex and expensive plant-derived products, especially in pharmaceutical production. Problems with up-scaling, low yields and high-priced process conditions result in an increased demand for models to provide comprehension, simulation, and optimization of production processes. In the last 25 years, many models have evolved in plant biotechnology; the majority of them are specialized models for a few selected products or nutritional conditions. In this article we review, delineate, and discuss the concepts and characteristics of the most commonly used models. Therefore, the authors focus on models for plant suspension and submerged hairy root cultures. The article includes a short overview of modeling and mathematics and integrated parameters, as well as the application scope for each model. The review is meant to help researchers better understand and utilize the numerous models published for plant cultures, and to select the most suitable model for their purposes.