Sugarcane (Saccharum X officinarum): A Reference Study for the Regulation of Genetically Modified Cultivars in Brazil

Global interest in sugarcane has increased significantly in recent years due to its economic impact on sustainable energy production. Sugarcane breeding and better agronomic practices have contributed to a huge increase in sugarcane yield in the last 30 years. Additional increases in sugarcane yield are expected to result from the use of biotechnology tools in the near future. Genetically modified (GM) sugarcane that incorporates genes to increase resistance to biotic and abiotic stresses could play a major role in achieving this goal. However, to bring GM sugarcane to the market, it is necessary to follow a regulatory process that will evaluate the environmental and health impacts of this crop. The regulatory review process is usually accomplished through a comparison of the biology and composition of the GM cultivar and a non-GM counterpart. This review intends to provide information on non-GM sugarcane biology, genetics, breeding, agronomic management, processing, products and byproducts, as well as the current technologies used to develop GM sugarcane, with the aim of assisting regulators in the decision-making process regarding the commercial release of GM sugarcane cultivars

Inibidores de proteinase de sementes de Bauhinia variegata : caracterização fisico-quimica e atividade biologica

Orientadores: Sergio Maramgani, Claudio Augusto Machado SampaioTese (doutorado) - Universidade Estadual de Campinas, Instituto de BiologiaResumo: A presença de inibidores de serinoproteinases foi investigada em sementes de duas variedades de Bauhinia variegata (Legumonosae, Caesalpinoideae). A purificação dos inibidores de Bauhínia variegata variedade cândida e variedade lilás por cromatografia de troca iônica, cromatografias de exclusão molecular e subsequente cromatografia de fase reversa, evidenciaram nestas espécies 3 isoformas (BvcTI-1, 2 e 3; e BvITI-1, 2 e 3) de inibidores de proteinase com cadeia polipeptídica única. A análise global de aminoácidos resultou em 167 e 180 resíduos de aminoácidos para BvcTI-3 e BvITI-3, respectivamente, e em massa molecular calculada de 18529 para BvcTI-3 e de 20018 para BvITI-3, e 4 resíduos de cisteína para BvcTI-3 e 2 resíduos de cisteína para BvITI-3. Estes resultados incluem estes inibidores na Família' de inibidores tipo Kunitz. A coloração para inibidores de tripsina após focalização isoelétrica demonstrou a presença de inibidores com pontos isoelétricos aparentes de 4,85, 5,00 e 5,15. A determinação da estrutura primária completa de BvcTI-3 e da estrutura primária do N-terminal das isoformas de BvcTI e BvITI, confiram alta homologia com inibidores tipo Kunitz. Os inibidores BvcTI e BvlTI atuam seletivamente sobre serinoproteinases. Extratos de sementes e inibidores purificados de Bauhínia variegata var. cândida e varo. lilás foram testados para atividade de inibição contra tripsina e quimotripsina bovina, porcina e humana, sendo que estas enzimas foram fortemente inibidas pelos inibidores em estudo. O teste de edema através do extravasamento de plasma em pele de coelhos, mostrou que BvcTI potenciou a calicreína de pâncreas de porcoAbstract: The presence of serine proteinase inhibitors was investigated in the seeds of Bauhinia variety cândida and Bauhinia variegata variety lilás (Leguminosae, Caesalpinoideae ). The purification of inhibitors from the Bauhinias by ion-exchange chromatography, molecular exclusion chromatography and subsequent reverse phase chromatography, showed the presence of three isoforms (BvcTI-1, 2 and 3 ; BvITI-1, 2 and 3) in the two species studied with single polypeptide chain. The aminoacid analisis of the forms BvcTI-3 and BvITI-3 resulted in 167 and 180 aminoacids residues, respectively, and the calculated molecular masses were 18529 to BvcTI-3 and 20018 to BvITI-3. It showed 4 cysteine residues to BvcTI-3 and 2 cysteine residues to BvITI-3, and no free thiol groups. This results suggest that these proteins belongs to the Kunitz-type plant inhibitors family. Staining for trypsin inhibitors after isoelectric focusing showed the presence of inhibitors with isoeletric point about 4.85, 5.00 and 5.15. The primary structure sequence of BvcTI-3 was determined, confirming the inhibitor as Kunitz type. The inhibitors BvcTI and BvlTI act selectively on serine proteinases. Extracts from seeds and purifieds inhibitors of both Bauhinia variegata variety candida and Bauhinia variegata variety lilas were tested for inhibitory activity against trypsin and chymotrypsin, from cattle, pig and humans. This three enzymes were strongly inhibited by both inhibitors. The oedema test in rabits showed the BvcTI made the kallikrein of porcine pancreas more potent to plasma. extravasationDoutoradoBioquimicaDoutor em Ciências Biológica

Potencial de Invasão de eucalipto pelas sementes produzidas nos plantios comerciais

Over the past few years, the area of planted forests has expanded considerably in Brazil, reaching a level of 6.3 million hectares, which represents approximately 0.7% of the country, mainly with some species of the Eucalyptus genus. However, due to the global concern with biological invasions by exotic species, risk assessment is necessary to evaluate the potential of Eucalyptus species escaping cultivated stands and invading surrounding native ecosystems. Seedlings by natural regeneration from Eucalyptus grandis and E. urophylla, pure and hybrids between the two species were evaluated, by assessing natural regeneration of Eucalyptus in 18 sites located in Brazilian states with plantation forest tradition (Minas Gerais, São Paulo, Rio Grande do Sul and Espírito Santo). Sample plots were installed in the planted stands, along forest roads and in a nearby pasture or native vegetation.,at different distances from the Eucalyptus stands. Regeneration of Eucalyptus was not recorded in 90% of the plots installed outside the eucalipt stands. However, seedlings of eucalypts were observed in 29% of the plots within the commercial stands (mean density of 37 seedlings per hectare) and 40% of the plots located along forest roads (157 seedlings per hectare on average), but no adult plants by natural regeneration were found. The seedlings of eucalypt species studied rarely were found beyond the limits of the cultivated areas. In the planted stands or along forest roads, eucalypts can germinate, but the seedlings apparently do not establish. So the potential for invasion by seeds produced in commercial plantations of Eucalyptus is low.Durante os últimos anos vem ocorrendo expansão na área das florestas plantadas no Brasil, atingindo o patamar de 6.310.000 hectares, que corresponde a aproximadamente 0,7% do território nacional, com destaque para algumas espécies do gênero Eucalyptus. No entanto, mediante a preocupação global com invasões biológicas por espécies exóticas, faz-se necessária a avaliação do risco de que espécies de eucalipto venham a invadir ecossistemas naturais a partir dos talhões de cultivo. Esta avaliação foi o objetivo do presente estudo, em diferentes regiões, com Eucalyptus grandis, E. urophylla, puros ou híbridos entre as duas espécies. O levantamento foi realizado em 18 áreas, localizadas em diferentes estados brasileiros com tradição florestal (MG, SP, RS e ES), sendo avaliada a regeneração em situações distintas: dentro da área de plantio (talhão), no carreador e em diferentes distâncias do plantio comercial (até 200 m), ocupada por pastagem ou por vegetação nativa. Observou-se que em 90% das parcelas analisadas fora das áreas de cultivo não ocorrem plantas ou plântulas de eucalipto. No entanto, plântulas de eucalipto foram observadas em 29% das parcelas dentro dos talhões comerciais (densidade média de 37 plântulas por hectare) e em 40 % das parcelas localizadas nos carreadores (157 plântulas por hectare, em média), mas não foram observadas plantas adultas oriundas da regeneração natural. Com este levantamento foi verificado que plântulas das espécies de eucalipto estudadas raramente são encontradas fora dos talhões de cultivo. Dentro dos talhões ou nos carreadores as espécies podem germinar, mas aparentemente as plântulas não se estabelecem. Considera-se baixo, portanto, o potencial de invasão pelo eucalipto por meio das sementes produzidas nos plantios comerciais

Purification And Characterization Of A Lectin From Crotalaria Paulina Seeds.

A lectin was purified from Crotalaria paulina seeds by ion-exchange and FPLC molecular exclusion chromatography. CrpL had an apparent molecular mass of 30 kDa, as determined by SDS-PAGE under non-reducing and reducing conditions. CrpL effectively agglutinated human and cow erythrocytes, and this activity was not affected by 20 mM EDTA, showing no dependence of divalent cations. Hemagglutination was inhibited by N-acetyl-D-galactosamine, D-galactose and was also inhibited by glycoproteins, fetuin and asialofetuin. The N-terminal amino acid sequence of CrpL was identical to those of other lectins from the genus Crotalaria, and amino acid composition showed high amounts of Asx and Glx, and was rich in Gly, Ala and Ser, as also reported for lectins from other Crotalaria species. CrpL inhibited the growth of Xanthomonas axonopodis pv. phaseoli and Xanthomonas axonopodis pv. passiflorae, suggesting a role of this lectin in the defense of seeds against bacterial infections.23437-4

Avaliação de quatro detergentes utilizados para solubilização de proteínas de membrana de Xylela fastidiosa empregando eletroforese bidimensional

Four different detergents, ASB 14, SB 3-10, CHAPS and Triton X100, were utilized to determine the optimal detergent for the solubilization of membrane proteins from the phytopathogenic bacterium Xylella fastidiosa. These proteins were differentially solubilized in distinct buffers containing the detergent and subjected to bidimensional electrophoresis within the non-linear pH range of 3-10. The detergents ASB 14 and SB 3-10 were the most effective revealing 221 and 157 spots, respectively. CHAPS and Triton X100 were less effective and revealed only 72 and 43 spots, respectively. MALDI-TOF tryptic peptide mass fingerprinting of 18 excised proteins from the ASB 14 treatment revealed that 83% were membrane proteins and that the theoretical efficiency of solubilization for ASB 14 was estimated to be 87%. This study demonstrates the effectiveness of the detergent ASB 14 for the solubilization of membrane proteins from the bacterium X. fastidiosa.O objetivo deste trabalho foi comparar a eficiência da solubilização de quatro detergentes comercialmente disponíveis, ASB 14, SB 3-10, CHAPS e Triton X100, na extração de proteínas de membrana da bactéria Xylella fastidiosa para estudos proteômicos. Estas proteínas foram solubilizadas em duas etapas em tampões diferenciados pelos detergentes e submetidas à eletroforese bidimensional (2-DE) em uma faixa de pH não linear de 3-10. Os detergentes ASB 14 e SB 3-10 foram os mais efecientes, revelando 221 e 157 proteínas, respectivamente, enquanto que o CHAPS e o Triton X100 resultaram somente 72 e 43 proteínas, respectivamente. A identificação das proteínas foi feita por 'peptide mass fingerprinting' em espectrometria de massa MALDI-TOF, através de peptídeos obtidos por digestão com tripsina in gel. Os 18 spots de proteínas do gel com tratamento com ASB 14 mostrou que 83% eram proteínas de membrana. Este estudo concluiu que o detergente ASB-14 foi o mais eficiente na solubilização de proteínas de membrana de Xylella fastidiosa.26927

The complete amino acid sequence of a trypsin inhibitor from Bauhinia variegata var. candida seeds

Trypsin inhibitors of two varieties of Bauhinia variegata seeds have been isolated and characterized. Bauhinia variegata candida trypsin inhibitor (BvcTI) and B. variegata lilac trypsin inhibitor (BVlTI) are proteins with M-r of about 20,000 without free sulfhydryl groups. Amino acid analysis shows a high content of aspartic acid, glutamic acid, serine, and glycine, and a low content of histidine, tyrosine, methionine, and lysine in both inhibitors. Isoelectric focusing for both varieties detected three isoforms (pI 4.85, 5.00, and 5.15), which were resolved by HPLC procedure. The trypsin inhibitors show K-i values of 6.9 and 1.2 nM for BvcTI and BvlTI, respectively. The N-terminal sequences of the three trypsin inhibitor isoforms from both varieties of Bauhinia variegata and the complete amino acid sequence of B. variegata var. candida L. trypsin inhibitor isoform 3 (BvcTI-3) are presented. The sequences have been determined by automated Edman degradation of the reduced and carboxymethylated proteins of the peptides resulting from Staphylococcus aureus protease and trypsin digestion. BvcTI-3 is composed of 167 residues and has a calculated molecular mass of 18,529. Homology studies with other trypsin inhibitors show that BvcTI-3 belongs to the Kunitz family. The putative active site encompasses Arg (63)-Ile (64).UNIFESP, Escola Paulista Med, Dept Biochem, BR-04044900 Sao Paulo, BrazilTech Univ Munich, Inst Lebensmittelchem, D-85748 Garching, GermanyDeutsch Forsch Anstalt Lebensmittelchem, D-85748 Garching, GermanyUniv Estadual Campinas, Inst Biol, Dept Biochem, BR-13083970 Campinas, SP, BrazilUNIFESP, Escola Paulista Med, Dept Biochem, BR-04044900 Sao Paulo, BrazilWeb of Scienc

Uso de ingredientes provenientes de OGM em rações e seu impacto na produção de alimentos de origem animal para humanos Use of ingredients from OGM in feed and its impact on the production of food of animal origin for human

Desde os primórdios o homem buscou selecionar as plantas alimentícias para maior produtividade. O conhecimento da estrutura do DNA permitiu que a engenharia genética se desenvolvesse consideravelmente fornecendo ferramentas para a realização de alterações específicas no genoma. Os produtos destas alterações são denominados transgênicos ou organismos geneticamente modificados (OGM) e apresentam alto potencial de aplicação em diversas áreas da atividade humana como: agricultura, medicina, saúde, produção e processamento de alimentos, produção bioquímica, controle de doenças e biorremediação. Atualmente, as plantas transgênicas, oriundas da tecnologia do DNA recombinante, trouxeram novas variedades já cultivadas em mais de 100 milhões de hectares em 23 países, incluindo o Brasil, onde 8 variedades já foram aprovadas pela Comissão Técnica Nacional de Biossegurança (CTNBio). Esse método de melhoramento genético facilitou a introdução de características desejáveis em plantas, como resistência a estresses bióticos e abióticos e otimização da composição de alguns nutrientes essenciais à saúde animal e humana. Enquanto estes avanços da biotecnologia abrem novas perspectivas para a solução de problemas em áreas como a agricultura, a liberação de transgênicos para uso na natureza traz preocupações quanto a possíveis problemas de natureza ecológica e para a saúde humana e animal. Estas preocupações deram origem à criação de agências governamentais para controlar o uso desta tecnologia e regulamentar a segurança dos alimentos transgênicos e seus derivados. Até o momento, os estudos científicos mostram que os transgênicos liberados comercialmente são tão seguros ou mais ao meio ambiente e a saúde animal e humana que os convencionais.<br>From the origins the man has looked and selected vegetables with nutritive value for larger productivity. The knowledge of DNA structure allowed genetic engineering to develop and supplying tools for the accomplishment of specific alterations in the genome considerably. The products of these alterations are denominated transgenic or organisms genetically modified (OGM) and they present high application potential in several areas of the human activity as: agriculture, medicine, health, production and processing of foods, biochemical production and control of diseases. Nowadays, transgenic plants, originating from technology of the DNA recombinant, brought new varieties cultivated already in more than 100 million hectares in 23 countries, including Brazil, where 8 varieties were already approved for the National Technical Commission of Biosafety (CTNBio). That method of genetic improvement facilitated the introduction of desirable characteristics in plants, such as, resistance to biotic and non-biotic stress and optimization of the composition of some essential nutrients to the animal and human health. While these progresses of the biotechnology open new perspectives for the solution of problems in areas as the agriculture, the transgenic liberation for use in the nature brings concerns as for possible problems of ecological nature and for the human and animal health. These concerns brought the creation of government agencies to control the use of this technology and to regulate the safety of transgenic foods and yours derived. Until the moment, the scientific studies show that the transgenic liberated commercially are so safe or more to the environment and the animal and human health that the conventional ones