Purificação, caracterização e análise da atividade bioinseticida, de um inibidor de tripsina em sementes de catanduva (Piptadenia moniliformis)

One Kunitz-type trypsin inhibitors (PmTI) was purified from Piptadenia moniliformis seeds, a tree of the sub-family Mimosoideae, by TCA precipitation, affinity chromatography on immobilized trypsin-Sepharose, DEAE cellulose (ion exchange) and Superose 12 (molecular exclusion) column FPLC/AKTA. The inhibitor has Mr of 25 kDa by SDS-PAGE and chromatography molecular exclusion. The N-terminal sequence of this inhibitor showed high homology with other family Kunitz inhibitors. This also stable variations in temperature and pH and showed a small decrease in its activity when incubated with DDT in the concentration of 100mM for 120 minutes. The inhibition of trypsin by PmTI was competitive, with Ki of 1.57 x10-11 M. The activity of trypsin was effectively inhibited by percentage of inhibition of 100%, among enzymes tested, was not detected inhibition for the bromelain, was weak inhibitor of pancreatic elastase (3.17% of inhibition) and inhibited by 76.42% elastase of neutrophils, and inhibited in a moderate, chymotrypsin and papain with percentage of inhibition of 42.96% and 23.10% respectively. In vitro assays against digestive proteinases from Lepidoptera, Diptera and Coleoptera pests were carried out. Several degrees of inhibition were found. For Anthonomus grandis and Ceratitis capitata the inhibition was 89.93% and 70.52%, respectively, and the enzymes of Zabrotes subfasciatus and Callosobruchus maculatus were inhibited by 5.96% and 9.41%, respectively, and the enzymes of Plodia. interpunctella and Castnia licus were inhibited by 59.94% and 23.67, respectively. In vivo assays, was observed reduction in the development of larvae in 4rd instar of C. capitata, when PmTI was added to the artificial diet, getting WD50 and LD50 of 0.30% and 0.33%, respectively. These results suggest that this inhibitor could be a strong candidate to plant management programs cross transgenic

Superação de dormência de sementes de Piptadenia moniliformis Benth

O angico-de-bezerro (Piptadenia moniliformis Benth.) é uma espécie arbórea melífera, de crescimento rápido, característica da caatinga do nordeste brasileiro, onde é muito abundante e com dispersão contígua e irregular. Seus ramos finos, junto com as folhas, servem como alimento para animais. Como suas sementes apresentam dormência por impermeabilidade do tegumento à água, objetivou-se neste trabalho avaliar a eficiência de tratamentos pré-germinativos para superar a dormência de suas sementes visando maximizar e uniformizar o processo de germinação. Foram estudados 28 tratamentos pré-germinativos: imersão em água a 70, 80, 90 ºC e fervente por 1, 2, 3, 4 e 5 minutos e imersão em ácido sulfúrico concentrado por 1, 5, 10, 15, 20, 25 e 30 minutos, além da testemunha. A semeadura foi realizada sobre papel em temperatura alternada de 20-30 ºC, utilizando-se quatro repetições de 25 sementes. Foram avaliadas as porcentagens de germinação das sementes considerando a protrusão da raiz primária, de plântulas normais e de sementes duras e o índice de velocidade de germinação. O delineamento experimental utilizado foi o inteiramente casualizado e as médias comparadas pelo teste de Scott-Knott a 5% de probabilidade. Os tratamentos de imersão em água, independentemente da temperatura e do período de imersão, aumentaram a porcentagem de germinação das sementes, comparativamente ao tratamento controle, mas foram menos eficientes que os tratamentos em ácido sulfúrico nos maiores períodos de imersão. Desta forma, a imersão por 20, 25 e 30 minutos em ácido sulfúrico é recomendada para a superação da dormência de sementes de Piptadenia moniliformis Benth.

Temperatura e substrato para a germinação de sementes de Piptadenia moniliformis Benth.

In germination tests, substrate and ideal temperature are important for providing favorable conditions for the germination of seeds and the eventual development of seedlings. This research aimed at evaluating the effects of temperature and substrate on the germination of Piptadenia moniliformis Benth seeds. Two trials were done. In the first, seeds were submitted to germination under constant temperatures of 5 to 40 degrees C (increases of 5 degrees C) and alternated temperatures of 20-25, 25-30, 30-35, 20-30, 25-35 and 20-35 degrees C on paper. In the second trial, temperatures of 25, 20-30 e 20-35 degrees C were used in the substrates on (OP) and between paper (BP), on (OS) and between sand (BS), on (0V) and between vermiculite (BV) and paper roll (PR). The following traits were evaluated: percentage of seeds with protrusion of primary root and normal seedlings; speed germination index; length of primary root and of the hypocotyls and the dry mass of seedlings. A completely randomized design was used with 14 treatments. In the first experiment and a factorial scheme of 7 x 3 (7 substrates and 3 temperatures) was used in the second with four replicates of 25 seeds each. In the first experiment the constant temperatures of 25 e 30 degrees C and the alternated of 20-30 e 20-35 degrees C provided best results for the germination of seeds. In the second, the temperature of 25 C associated with the substrates BP, BS and BV surpassed the other treatments proving adequate for carrying out the germination tests of Piptadenia moniliformis Benth seeds.

Viabilidade de sementes de Piptadenia moniliformis Benth. Pelo teste de tetrazólio

Piptadenia moniliformis Benth. is a tree species which is important for beekeeping, as well as being recommended for soil restoration, reforestation, wood production for small civil construction projects, and cattle and sheep forage. Information on how to evaluate seed physiological quality is still scarce and in this was the study aimed to adapt the procedures of the tetrazolium test to assess the viability of P. moniliformis seeds. Four seed lots were scarified in sulphuric acid for 30 min, and soaked between paper towels at 25 °C for 24 hours. The seed coat was then removed and the naked seeds immersed in tetrazolium solutions with concentrations of 0.05, 0.075 and 0.1% for 2, 3, and 4 hours at 35 oC in the dark. Each treatment consisted of four replications of 25 seeds. The embryos were classified according to viability based on the staining patterns. The previous soaking of the seeds for 24 hours at 25 oC between paper towels, followed by the removal of the seed coat and staining of the naked seeds for 4 hours in a 0.075% tetrazolium solution at 35 oC was the most efficient method for evaluating the viability of P. moniliformis Benth seeds.

Qualidade fisiológica de sementes de Piptadenia moniliformis Benth

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

GERMINAÇÃO DE SEMENTES DE Piptadenia moniliformis Benth. SOB ESTRESSE HÍDRICO

O período de germinação e o estabelecimento de plântulas é um dos fatores mais importantes para a sobrevivência das espécies, principalmente nos locais em que a disponibilidade de água é limitada, como na região da Caatinga. Neste sentido, o objetivo deste trabalho foi avaliar o efeito do estresse hídrico sobre a germinação de sementes de Piptadenia moniliformis Benth. Foram utilizados três lotes (L1, L2 e L3), correspondentes aos anos de produção de 2006, 2007 e 2008, respectivamente. Antes do teste de germinação, as sementes foram escarificadas com ácido sulfúrico concentrado durante 30 minutos. Para induzir o deficit hídrico, foi utilizado o polietileno glicol (PEG 6000), nos seguintes potenciais osmóticos: - 0,3; -0,6; -0,9, -1,2 e -1,5 MPa e a água (0 MPa) sob as temperaturas de 25 e 30ºC. As características avaliadas foram: porcentagem de germinação e de plântulas normais, índice de velocidade de germinação e massa seca de plântulas. O processo germinativo de sementes de Piptadenia moniliformis Benth. é comprometido a partir de potenciais hídricos inferiores a -0,6 MPa a 25 e 30 °C; potenciais hídricos iguais ou inferiores a -1,2 MPa inibem a formação de plântulas normais nas duas temperaturas; a tolerância ao estresse hídrico simulado com PEG 6000 é variável entre lotes de sementes e temperaturas de germinação.

Purificação, caracterização e análise da atividade bioinseticida, de um inibidor de tripsina em sementes de catanduva (Piptadenia moniliformis)

One Kunitz-type trypsin inhibitors (PmTI) was purified from Piptadenia moniliformis seeds, a tree of the sub-family Mimosoideae, by TCA precipitation, affinity chromatography on immobilized trypsin-Sepharose, DEAE cellulose (ion exchange) and Superose 12 (molecular exclusion) column FPLC/AKTA. The inhibitor has Mr of 25 kDa by SDS-PAGE and chromatography molecular exclusion. The N-terminal sequence of this inhibitor showed high homology with other family Kunitz inhibitors. This also stable variations in temperature and pH and showed a small decrease in its activity when incubated with DDT in the concentration of 100mM for 120 minutes. The inhibition of trypsin by PmTI was competitive, with Ki of 1.57 x10-11 M. The activity of trypsin was effectively inhibited by percentage of inhibition of 100%, among enzymes tested, was not detected inhibition for the bromelain, was weak inhibitor of pancreatic elastase (3.17% of inhibition) and inhibited by 76.42% elastase of neutrophils, and inhibited in a moderate, chymotrypsin and papain with percentage of inhibition of 42.96% and 23.10% respectively. In vitro assays against digestive proteinases from Lepidoptera, Diptera and Coleoptera pests were carried out. Several degrees of inhibition were found. For Anthonomus grandis and Ceratitis capitata the inhibition was 89.93% and 70.52%, respectively, and the enzymes of Zabrotes subfasciatus and Callosobruchus maculatus were inhibited by 5.96% and 9.41%, respectively, and the enzymes of Plodia. interpunctella and Castnia licus were inhibited by 59.94% and 23.67, respectively. In vivo assays, was observed reduction in the development of larvae in 4rd instar of C. capitata, when PmTI was added to the artificial diet, getting WD50 and LD50 of 0.30% and 0.33%, respectively. These results suggest that this inhibitor could be a strong candidate to plant management programs cross transgenic

Molecular characterization and fumonisin production by Fusarium verticillioides isolated from corn grains of different geographic origins in Brazil

Gibberella moniliformis is most commonly associated with maize worldwide and produces high levels of fumonisins, some of the most agriculturally important mycotoxins. Studies demonstrate that molecular methods can be helpful for a rapid identification of Fusarium species and their levels of toxin production. The purpose of this research was to apply molecular methods (AFLP, TEF-1 alpha partial gene sequencing and PCR based on MAT alleles) for the identification of Fusarium species isolated from Brazilian corn and to verify if real time RT-PCR technique based on FUM1 and FUM19 genes is appropriated to estimate fumonisins B(1) and B(2) production levels. Among the isolated strains, 96 were identified as Fusarium verricillioides, and four as other Fusarium species. Concordant phylogenies were obtained by AFLP and TEF-1 alpha sequencing, permitting the classification of the different species into distinct clades. Concerning MAT alleles, 70% of the F. verricillioides isolates carried the MAT-1 and 30% MAT-2. A significant correlation was observed between the expression of the genes and toxin production r=0.95 and r=0.79 (correlation of FUM1 with FB(1) and FB(2), respectively, P < 0.0001): r=0.93 and r =0.78 (correlation of FUM19 with FB(1) and FB(2). respectively, P < 0.0001). Molecular methods used in this study were found to be useful for the rapid identification of Fusarium species. The high and significant correlation between FUM1 and FUM19 expression and fumonisins production suggests that real time RT-PCR is suitable for studies considering the influence of abiotic and biotic factors on expression of these genes. This is the first report concerning the expression of fumonisin biosynthetic genes in Fusarium strains isolated from Brazilian agricultural commodity. (c) 2010 Elsevier B.V. All rights reserved.

Diatom deposits in the ostpreußischen Haffe

Als man nach dem ersten Weltkrieg im verkleinerten Deutschland nach der Möglichkeit von Neulandgewinnung suchte, dachte man auch an eineTrockenlegung der ostpreußischen Haffe. Aus diesem Anlaß wurden umfangreiche Bohrungen ausgeführt, um ein möglichst genaues Bild vom Untergrunde der Haffe zu bekommen. Auf Veranlassung der Preußischen Geologischen Landesanstalt wurde ich mit der Untersuchung der Diatomeen in den Bohrproben beauftragt. Die Arbeit wurde 1934 begonnen und Ende 1937 wurde der letzte Arbeitsbericht abgeliefert. Die beabsichtigte Veröffentlichung ist bisher unterblieben, weil die Druckvorlagen später verloren gegangen sind. Seitdem sind über die Haffuntersuchungen mehrere Teilergebnisse veröffentlicht worden, von denen hier schon wegen der Terminologie die pollenanalytischen Arbeiten von L. HEIN (1941) und HUGO GROSS (1941) erwähnt seien, auf die im Abschnitt Il 2e näher eingegangen wird. Bei der geologischen Auswertung war Zurückhaltung geboten; denn es wäre gewagt, allein aus der Perspektive der Diatomeenforschung endgültige Aussagen machen zu wollen. Darum habe ich mich bemüht, das Material so weit aufzuschließen, daß es Geologen später auch bei veränderter Fragestellung auswerten können. "Die Theorien wechseln, aber die Tatsachen bleiben." Der Initiative des Herrn Prof. Dr. K. GRIPP und der finanziellen Hilfe der Deutschen Forschungsgemeinschaft ist es zu verdanken, daß die vorliegende Arbeit im Druck erscheinen kann. Zusammenfassung 1. Nur in den alluvialen Schichten des Kurischen Haffs wurden Diatomeen gefunden. 2. Die Diatomeenflora des Kurischen Haffs besteht zur Hauptsache aus Süßwasserformen. 3. Salzwasserformen finden sich in allen Schichten verstreut unter der Süßwasserflora. Wenn sie auch nach Zahl der Arten in manchen Proben einen erheblichen Prozentsatz der Flora ausmachen, so ist doch die Zahl der Individuen stets so gering, daß man nirgends von einer Brackwasserflora sprechen kann. 4. Die Süßwasserflora besteht in den unteren Schichten vorwiegend aus Grundformen; und zwar machen die epiphytischen Bewohner flacher Sumpfgewässer einen großen Teil der Flora aus. 5. In einzelnen Bohrungen kommt in den untersten alluvialen Schichten eine Grundflora mit zahlreichen Mastogloien vor. Dies sind die ältesten diatomeenführenden Schichten, entstanden in isolierten Sumpfgewässern. 6. Die übrigen Schichten mit überwiegender Grundflora sind vermutlich Ablagerungen der Ancyluszeit. 7. Die oberen Schichten, in denen die Planktondiatomeen überwiegen, dürften größtenteils der Litorina-Transgressionszeit angehören, jedoch ist der Transgressions-Kontakt nicht klar zu erkennen. 8. Das Ende der Litorinazeit ist noch weniger erkennbar, da eine grundsätzliche Veränderung der Flora nach oben nicht zu beobachten ist. 9. Die ostbaltischen Charakterformen sind in allen Schichten vertreten.

The Global Plankton Database: an inventory and data from the Former Soviet Union expeditions

At present time, there is a lack of knowledge on the interannual climate-related variability of zooplankton communities of the tropical Atlantic, central Mediterranean Sea, Caspian Sea, and Aral Sea, due to the absence of appropriate databases. In the mid latitudes, the North Atlantic Oscillation (NAO) is the dominant mode of atmospheric fluctuations over eastern North America, the northern Atlantic Ocean and Europe. Therefore, one of the issues that need to be addressed through data synthesis is the evaluation of interannual patterns in species abundance and species diversity over these regions in regard to the NAO. The database has been used to investigate the ecological role of the NAO in interannual variations of mesozooplankton abundance and biomass along the zonal array of the NAO influence. Basic approach to the proposed research involved: (1) development of co-operation between experts and data holders in Ukraine, Russia, Kazakhstan, Azerbaijan, UK, and USA to rescue and compile the oceanographic data sets and release them on CD-ROM, (2) organization and compilation of a database based on FSU cruises to the above regions, (3) analysis of the basin-scale interannual variability of the zooplankton species abundance, biomass, and species diversity.