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Las tripsinas son las principales peptidasas digestivas de crustáceos. En los últimos años se han producido revisiones sobre las tripsinas de invertebrados, así como de crustáceos, e incluso de las presentes en determinada especie de crustáceo. Sin embargo, los aspectos relacionados con la purificación de estas enzimas (y sus isoformas) no se han abordado en estas revisiones. En la presente revisión se exponen los procedimientos más empleados en la purificación de tripsinas de crustáceos, las dificultades en la separación de isoformas, y se discuten las experiencias de los autores en la purificación de isoformas de tripsina en la langosta espinosa Panulirus argus, las cuales pudieran ser de interés para los que se inicien en el campo de la purificación de enzimas tipo tripsina de crustáceos. Se excluyen los aspectos básicos relacionados con los principios de cada método y con la purificación de proteínas en general, los cuales pueden ser encontrados en la vasta literatura publicada tanto en revisiones como en libros especializados.

At higher temperatures, lettuce seeds may not germinate, resulting in problems for crop establishment in the field and production. This study aimed to evaluate the maternal effect for thermotolerance on lettuce seeds and find enzymatic markers. We used the thermoinhibition tolerant cultivar 'Everglades', the sensitive 'Verônica', their reciprocal hybrids, F1 and F2 generation. Seeds were submitted to germination and enzymatic tests (catalase, superoxide dismutase, ascorbate peroxidase, and endo-ß-mannanase). Germination (first and final counting) and speed index were compared through Tukey's test. 'Everglades' and its reciprocal hybrids, have not been influenced by temperature regarding germination compared to the other treatments. Also, the maternal effect was observed for the F1 generation. Higher temperatures interfere with the activity of all enzymes evaluated, consequently in the physiological parameters. However, superoxide dismutase and endo-ß-mannanase have shown direct and more expressive correlation with changes on the physiological parameters compared to the other enzymes. Thus, it is possible to conclude that those enzymes can be used as markers for thermotolerant genotype selection. [ABSTRACT FROM AUTHOR]

Zaprionus indianus (fig fly) is a drosophilid of African origin that was recorded in Brazilian territory in 1999, quickly dispersing throughout the country, causing impacts on fig culture, and becoming one of the dominant Drosophilidae species in the Cerrado biome. Due to the fact it is an invasive species and a potential pest in fruit cultivars, it is essential to monitor the variations in the populations of this species aiming to verify its genetic and demographic structure and possible impacts on the drosophilid community. The demography and the genetic data obtained from the Est3 polymorphism of fig fly have been evaluated in six populations of Brazilian Cerrado sampled during the dry season. Multivariate analyses of PCA, UPGMA and landscape genetics were performed. The results have indicated variation in the demography of the sampled populations, the presence of two alleles (Est31 and Est34) in all of them, and a higher isolation degree of the Guaíra (SP) population. Thus, the fig fly is a colonizer of the Brazilian Cerrado, and the polymorphisms of Est3 locus are an appropriate genetic marker to investigate the bioinvasion process. [ABSTRACT FROM AUTHOR]

Coffee seeds are sensitive to desiccation and are used or stored with different moisture content values, which may affect the results of quality assessment. The aim of this study was to evaluate the changes in protein profile in electrophoresis gel in coffee seeds with different moisture content values under freeze-drying and without freeze-drying. Two lots of arabica coffee seeds were used, one of newly-harvested seeds and another of stored seeds. The seeds were dried to the moisture content values of 12, 15, 20, 25, 30, 35, and 40%. The physiological quality of the seeds was assessed through the germination test, electrical conductivity, and the profiles of the enzymes SOD, CAT, PO, GOT, MDH, and EST and of heat-resistant proteins. In general, there is an effect on expression of these enzymes in accordance with the presence of free water in the seeds. Moist seeds have little to no enzyme expression. The freeze-drying process allows preservation of coffee seed quality and does not change the functionality of the enzymes studied. The isoenzyme profiles of the antioxidant process in arabica coffee seeds are affected by the initial moisture content of the seeds. The freeze-drying process of the seeds ensures greater sensitivity in detection of the expression of these isoenzymes. [ABSTRACT FROM AUTHOR]

A demanda por sementes de hortaliças com alta qualidade e a obtenção de mudas vigorosas tem aumentado nos últimos anos, pois estas características constituem fatores determinantes do êxito da produção. As hortaliças têm uma importância nacional crescente, dentre elas se destaca a couve, por ser importante fonte de renda para pequenos agricultores. Objetivou-se adequar a metodologia do teste de envelhecimento acelerado tradicional e com solução saturada de NaCl em sementes de couve e avaliar a atividade enzimática após o teste de vigor. Foram utilizados seis lotes de sementes de couve e determinou-se o grau de umidade, peso de mil sementes, primeira contagem de germinação, germinação, índice de velocidade de germinação, emergência, estande inicial e índice de velocidade de emergência. No teste de envelhecimento acelerado, as sementes foram submetidas ao método tradicional e com solução saturada de NaCl, por períodos de 0; 24; 48; 72 e 96 horas. Realizou-se também a análise eletroforética das isoenzimas superóxido dismutase (SOD), catalase (CAT) e álcool desidrogenase (ADH). O teste de envelhecimento acelerado, utilizando o método tradicional na combinação de 72 horas a 41 °C é adequado para avaliar o potencial fisiológico de sementes de couve. As análises isoenzimáticas superóxido dismutase (SOD), catalase (CAT) e álcool desidrogenase (ADH) demonstram que os marcadores bioquímicos são eficientes e promissores na diferenciação de sementes de couve após o envelhecimento acelerado.

Copyright of Revista Mexicana de Ciencias Agrícolas is the property of Instituto Nacional de Investigaciones Forestales, Agricolas y Pecuarias and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

ABSTRACT We describe isoenzymes variability in six populations of Grevillea robusta from a provenances and progenies test established in a randomized block design with five plants per replication in Southern Brazil. The population genetic structure was examined by using biochemical markers in 5-year old trees, specifically at MDH-3, PGM-2, DIA-2, PO-1, PO-2, SOD-1, and SKDH-1 loci. The northern provenances (Rathdowney and Woodenbong) showed a strong divergence in relation to the average of provenances when alleles per locus (Ap), allele richness (Rs), Nei's gene diversity (H), and inbreeding coefficient (f) were considered. Inbreeding in varying degrees was detected. The commercial control showed the highest inbreeding coefficient, (f = 0.4448), whereas the provenance average was f = 0.2306, possibly due to insufficient sampling of populations in their origin (Australia). Despite its restricted natural range, a positive correlation between genetic divergence and geographic distance among original populations was detected. Genetic distance and cluster analyses based on the Bayesian model revealed three distinct provenance groups: 1) Rathdowney-QLD and Woodenbong-QLD; 2) Paddy's Flat-NSW; and 3) Mann River-NSW, Boyd River-NSW and the commercial control (material used in Brazil). The grouping of the control to both Mann River-NSW and Boyd River-NSW provenances suggests that the northern provenances have the highest potential for genetic improvement of wood productivity in Brazil, due to their high genetic diversity and low inbreeding coefficient.

Handroanthus serratifolius seeds are considered orthodox and have some mechanisms that assist in desiccation tolerance, like heat-resistant proteins and the presence of protective systems against free radical production. In order to investigate the drying effect on the quality of Handroanthus serratifolius seeds collected with 28% water content two methods were used: slow drying (30 °C) and fast drying (45 °C) until water contents equaled 20%, 15%, 10% and 5%. Seed quality was assessed by germination test, first count of germination, speed germination, emergence and speed emergence. The expression of heat-resistant proteins and enzymes catalase, esterase and isocitrate lyase was assessed by electrophoresis. The gene expression of sHSP17.5, CAT3 and ICL6 was quantified by real time PCR. Higher values of germination and vigor were obtained in seeds with 20% water content, subjected to fast drying. There was induction of heat-resistant proteins expression for both drying methods. There was variation in the electrophoretic pattern of catalase, esterase and isocitrate lyase in seeds subjected to different treatments. The gene expression in Handroanthus serratifolius seeds varies depending on methods of drying and water content of seeds. The drying of Handroanthus serratifolius seeds until the water content between 15 and 10% compromises their physiological quality.

Copyright of Biomédica: Revista del Instituto Nacional de Salud is the property of Instituto Nacional de Salud of Colombia and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The silicon utilization may result in the increased biological ability of the plants to withstand the harsh conditions of the environment. In this sense, the study aimed to evaluate the effect of the application of rice husk ash to the soil in rice plants under salt stress on isoenzymes expression. The experimental design was a randomized block in factorial A x B (Factor A- Five doses of rice husk ash applied to soil: 0.0, 500, 1000, 1500, and 2000 kg ha-1, Factor B- Three salt concentrations: 0, 4, and 8 mM), with four replications. Tests were the first count of germination and germination, and expression of isozyme systems glutamate oxaloacetate transaminase, alcohol dehydrogenase, glutamate dehydrogenase, and sorbitol. Variations in the all analyzed electrophoretic systems may be associated with salt stress caused during the growth and development of rice plants. The isoenzyme system glutamate oxaloacetate transaminase is a complementary tool to assess the physiological rice seed quality. [ABSTRACT FROM AUTHOR]

Se investigó la variación isoenzimática del arbusto raro Sophora koreensis en el condado de Yanggu (Corea del Sur), en el extremo sur de su área de distribución. Para obtener información sobre los procesos ecológicos a nivel del paisaje, se estudiaron dos o tres poblaciones para cada una de las cuatro localidades (en total, 10 poblaciones) en un radio de unos 6 km. Se encontraron bajos niveles de variación genética intra-poblacional (%P = 13,6, A = 1,14 y He = 0,026) y un grado moderado de diferenciación genética entre poblaciones (FST = 0,203). El análisis de la varianza molecular (AMOVA) reveló un porcentaje sustancialmente mayor de variación entre poblaciones dentro cada una de las localidades (17%) que entre localidades (5%). No hubo una relación significativa entre la divergencia genética y el logaritmo de la distancia geográfica entre pares de poblaciones (r = 0,032, P = 0,842). Estos resultados sugieren un flujo genético limitado entre poblaciones dentro de las localidades, además de indicar que la mejor estrategia para la preservación de la diversidad genética de S. koreensis es la conservación del máximo número de poblaciones posible.

The present study was carried out in Corpoica, C.I. Tibaitata (Mosquera, Colombia) at an altitude of 2,540 m a.s.l. in six environments composed of different seasons and at an altitude of 1,485 m a.s.l. in the International Center for Tropical Agriculture (CIAT ), Calima, Colombia. Morphological, physiological, biochemical, and molecular descriptors were used to estimate the genetic variability between 36 Colombian bean accessions, of which four were wild and the others cultivated. Diacol Calima (Nueva Granada from the Andean gene pool) and ICA Pijao (Mesoamerican from the mesoamerican gene pool) were used as controls. The combined analysis of the qualitative and quantitative variables was carried out with Gower distance and an unified data matrix with 315 descriptors. The relationships between the genetic distances differentiated the bean collection into two genetic groups: Andean and Mesoamerican. The following groups of characteristics presented high association: total morphology with qualitative morphology (P=0.91), physiological evaluation with grouped evaluation of morphological, quantitative, and physiological characteristics (P=0.91), characterization of isoenzymatic and molecular markers with respect to just molecular markers (P=0.99) and the characterization of all the studied markers in relation to the molecular and isoenzymatic markers (P=0.88).

Crescente interesse tem se estabelecido para a análise da diversidade genética de espécies Conyza bonariensis, C.canadensis e C.sumatrensis, popularmente conhecidas como buva ou voadeira, que nos últimos anos vêm causando vários prejuízos nas lavouras do Brasil e do mundo, principalmente nas plantações de soja. A proposta do presente estudo foi estimar a variabilidade genética de amostras de C.sumatrensis provenientes da região noroeste do Estado do Paraná. A análise de isozimas em tecidos de folhas das plantas de C. sumatrensis foi realizada para estimar a variabilidade genética dentro de cada população e entre populações diferentes, no sentido de recomendar um tratamento diferencial ou uniforme para o controle dessas plantas daninhas na referida região. Foram analisados quatro sistemas enzimáticos (ACP, GPI, MDH e PGM) e detectados 10 locos com 10 alelos, os quais não apresentaram diversidade genética dentro e entre as populações analisadas, comprovado pela presença de apenas indivíduos homozigotos. As enzimas analisadas no presente estudo indicaram que as plantas das três regiões são geneticamente uniformes, e a uniformidade genética verificada para os referidos locos é um indicativo prévio de que é possível utilizar doses equivalentes do glifosato para controlar o crescimento desses biótipos.

To evaluate the effect of accelerated aging to overcome dormancy of hybrid Brachiaria cv. Mulato II (U. brizantha x U. decumbens x U. ruziziensis), seeds were collected from two lots with and without scarification, submitted to 42 °C and high humidity (98%) for different periods (0, 24, 48, 72 and 96 h) and then dried at 30 °C to the humidity of 9.1%. The physiological changes were evaluated by tetrazolium, germination, electrical conductivity, respiratory activity and enzymatic evaluation. The exposure of the hybrid Brachiaria cv. Mulato II to accelerated aging at 42 °C and 98% relative air humidity for up to 96 hours does not alter its viability by the tetrazolium test. Accelerated aging at 42 °C and 98% of humidity for up to 48 hours favors the germination of the scarified seeds of hybrid Brachiaria cv. Mulato II, with increased expression of α-amylase enzyme and reduction of electrical conductivity values. Loss of vigor occurs in scarified seeds of hybrid Brachiaria cv. Mulato II when exposed to accelerated aging at 42 °C and 98% of humidity from 96 hours with consequent changes in enzymatic systems and increased of the respiratory rate. [ABSTRACT FROM AUTHOR]

Copyright of Fitosanidad is the property of Instituto de Investigaciones de Sanidad Vegetal and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Se determinó la estructura genética poblacional y sus implicancias dentro de la cadena trófica del copépodo Acartia lilljeborgii en la costa de Yucatán, mediante el uso de isoenzimas y su posterior determinación por electroforesis. Nueve sistemas se expresaron con un máximo de 30 loci. El polimorfismo y heterocigosis de las poblaciones estudiadas, al ser comparados con otras especies de copépodos, resultaron ser bajos y de los cuatro loci que presentaron variación, solo los loci EST2 y PGM1 se expresaron en todos los sitios de muestreo. La PGM1 presentó el mayor valor de heterocigosis (0,4728) y la GPI1 fue la de menor valor (0,3200). Ningún sistema se apartó de la condición de equilibrio de Hardy-Weinberg. Los valores de FST estuvieron entre los valores de -0,0798 para PGM1 y de 0,5899 para FBP1. Las tres poblaciones de A. lilljeborgii analizadas en la costa de Yucatán, presentaron una baja variabilidad y diferenciación genética entre las poblaciones, como lo indican los valores de distancia genética de 0,0585 que fue el mayor para las poblaciones de Chelem y de Río Lagartos, lo que sugiere que estas poblaciones están parcialmente aisladas. Sin embargo, debido a que A. lilljeborgii produce huevos diapáusicos o de reposo, que pueden ser transportados fuera o dentro de éstos cuerpos de agua, es probable que esta condición mantenga las frecuencias alélicas y genotípicas similares o muy cercanas entre sí. Los resultados mostraron la existencia de flujo genético, dado a través de una dispersión pasiva de los huevos diapáusicos que produce esta especie.

The study of the genetic structure of wild plant populations is essential for their management and conservation. Several DNA markers have been used in such studies, as well as isozyme markers. In order to provide a better comprehension of the results obtained and a comparison between markers which will help choose tools for future studies in natural populations of Oryza glumaepatula, a predominantly autogamous species, this study used both isozymes and microsatellites to assess the genetic diversity and genetic structure of 13 populations, pointing to similarities and divergences of each marker, and evaluating the relative importance of the results for studies of population genetics and conservation. A bulk sample for each population was obtained, by sampling two to three seeds of each plant, up to a set of 50 seeds. Amplified products of eight SSR loci were electrophoresed on non-denaturing polyacrylamide gels, and the fragments were visualized using silver staining procedure. Isozyme analyses were conducted in polyacrylamide gels, under a discontinuous system, using six enzymatic loci. SSR loci showed higher mean levels of genetic diversity (A=2.83, p=0.71, A P=3.17, Ho=0.081, He=0.351) than isozyme loci (A=1.20, p=0.20, A P=1.38, Ho=0.006, He=0.056). Interpopulation genetic differentiation detected by SSR loci (R ST=0.631, equivalent to F ST=0.533) was lower than that obtained with isozymes (F ST=0.772). However, both markers showed high deviation from Hardy-Weinberg expectations (F IS=0.744 and 0.899, respectively for SSR and isozymes). The mean apparent outcrossing rate for SSR ( =0.14) was higher than that obtained using isozymes ( =0.043), although both markers detected lower levels of outcrossing in Amazonia compared to the Pantanal. The migrant number estimation was also higher for SSR (Nm=0.219) than isozymes (Nm=0.074), although a small number for both markers was expected due to the mode of reproduction of this species, defined as mixed with predominance of self fertilization. No correlation was obtained between genetic and geographic distances with SSR, but a positive correlation was found between genetic and geographic distances with isozymes. We conclude that these markers are divergent in detecting genetic diversity parameters in O. glumaepatula and that microsatellites are powerful for detecting information at the intra-population level, while isozymes are more powerful for inter-population diversity, since clustering of populations agreed with the expectations based on the geographic distribution of the populations using this markerEl estudio de la estructura genética de poblaciones de plantas silvestres es esencial para su manejo y conservación. Varios marcadores de ADN e isoenzimas se han utilizado en este tipo de análisis. Con el fin de proporcionar una mejor comprensión de los resultados obtenidos y saber que marcador codominante elegir para futuros estudios en poblaciones naturales de Oryza glumaepatula, este trabajo busco evaluar y comparar dos marcadores de ADN, isoenzimas y microsatélites, en la diversidad y estructura genética de 13 poblaciones, destacando las similitudes y divergencias de cada marcador, así como la importancia relativa de los resultados en genética de poblaciones y conservación. Para los SSR, ocho loci SSR fueron evaluados, y los fragmentos se visualizaron utilizando el procedimiento de coloración con plata. Los análisis de isoenzimas se realizaron en geles de poliacrilamida, en los seis loci enzimáticos. Los loci SSR mostraron mayores niveles de diversidad genética que los loci isoenzimáticos, en promedio. La diferenciación genética entre los loci SSR (R ST=0.631, equivalente a F ST=0.533) fue inferior a la obtenida con las isoenzimas (F ST=0.772). Ambos marcadores mostraron alta desviación del equilibrio de Hardy-Weinberg (F IS=0.744 y 0.899, respectivamente, para SSR e isoenzimas). La tasa media aparente de cruzamiento para SSR ( =0.14) fue mayor que la obtenida con isoenzimas ( =0.043), aunque ambos marcadores detectaron niveles más bajos en la tasa de fecundación cruzada para la Amazonia, en comparación con la región del Pantanal. La estimación de número de migrantes también fue mayor para los SSR (Nm=0.219) que en isoenzimas (Nm=0.074). No se obtuvo ninguna correlación entre las distancias genéticas y geográficas para los SSR, y para las isoenzimas se obtuvo una correlación positiva entre las distancias genéticas y geográficas. Llegamos a la conclusión de que estos marcadores son divergentes en la detección de los parámetros de la diversidad genética en O. glumaepatula y que los microsatélites son más eficientes para detectar la información a nivel intra-poblacional, mientras que las isoenzimas son más potentes para detectar la diversidad entre poblaciones.

Hancornia speciosa Gomes is a fruit tree native from Brazil that belongs to Apocinaceae family, and is popularly known as Mangabeira. Its fruits are widely consumed raw or processed as fruit jam, juices and ice creams, which have made it a target of intense exploitation. The extractive activities and intense human activity on the environment of natural occurrence of H. speciosa has caused genetic erosion in the species and little is known about the ecology or genetic structure of natural populations. The objective of this research was the evaluation of the genetic diversity and genetic structure of H. speciosa var. speciosa. The genetic variability was assessed using 11 allozyme loci with a sample of 164 individuals distributed in six natural populations located in the States of Pernambuco and Alagoas, Northeastern Brazil. The results showed a high level of genetic diversity within the species (e= 0.36) seeing that the most of the genetic variability of H. speciosa var. speciosa is within its natural populations with low difference among populations ( or = 0.081). The inbreeding values within ( = -0.555) and among populations ( =-0.428) were low showing lacking of endogamy and a surplus of heterozygotes. The estimated gene flow ( m ) was high, ranging from 2.20 to 13.18, indicating to be enough to prevent the effects of genetic drift and genetic differentiation among populations. The multivariate analyses indicated that there is a relationship between genetic and geographical distances, which was confirmed by a spatial pattern analysis using Mantel test (r = 0.3598; p = 0.0920) with 1000 random permutations. The high genetic diversity index in these populations indicates potential for in situ genetic conservation.

The knowledge of genetic diversity patterns increase the efficiency of the conservation and the enrichment of the genetic resourses. This study allowed the discrimination of the existing genetic variability in a Colombian collection of shrub bean by phaseolin patterns and isoenzymatic markers. Bean seed proteins revelated that the phaseolin patterns types T and C are predominant in the Andean pool, type S in the Meso-American pool and type B in Colombian and Central American accessions, with a predominance of 81% of phaseolin T in the Andean pool, and 78% of phaseolin B in the Meso-American pool. The accessions of cultivated and wild beans showed variation in 10 of the studied enzymatic systems: αβ-EST , GOT, αβ-ACP, DIA , PRX, AS D, 6-PGDH, MDH, IDH and ME; and monomorphism in the PGI and PGM systems. The isozyme systems presented 19 bands of activity, of which 74% were polymorphic loci. Both in the Andean and Meso-American genetic pools, the loci Mdh-1, Mdh-2, β-Est-1, Skdh and Me exhibited polymorphisms. Single alleles in the Meso-American pool were found in 6-Pgdh-2(103), Mdh-1(100), Idh100, α-Est-1(100), α-Est-2(100), and Dia-195; and in the Andean pool, in 6-Pgdh-1(100), and Acp-2(100). For degree of domestication, the wild and cultivated accessions presented polymorphisms in 58 and 47% of the lOCi, respectively. The enzymatic relationship cluster analysis of the studied bean collection revealed three distinct groups of accessions; namely the Meso-American pool, including its cultivated and wild accessions; the Andean pool, which is mainly comprised of cultivated accessions, plus the wild DGD-626; and finally, featured by a high degree of enzymatic polymorphism and by the presence of the type I phaseolin, a third group that contains only a wild accession from the northern, Peruvian Andes

The enzyme chitinase from Moniliophthora perniciosa the causative agent of the witches' broom disease in Theobroma cacao, was partially purified with ammonium sulfate and filtration by Sephacryl S-200 using sodium phosphate as an extraction buffer. Response surface methodology (RSM) was used to determine the optimum pH and temperature conditions. Four different isoenzymes were obtained: ChitMp I, ChitMp II, ChitMp III and ChitMp IV. ChitMp I had an optimum temperature at 44-73ºC and an optimum pH at 7.0-8.4. ChitMp II had an optimum temperature at 45-73ºC and an optimum pH at 7.0-8.4. ChitMp III had an optimum temperature at 54-67ºC and an optimum pH at 7.3-8.8. ChitMp IV had an optimum temperature at 60ºC and an optimum pH at 7.0. For the computational biology, the primary sequence was determined in silico from the database of the Genome/Proteome Project of M. perniciosa, yielding a sequence with 564 bp and 188 amino acids that was used for the three-dimensional design in a comparative modeling methodology. The generated models were submitted to validation using Procheck 3.0 and ANOLEA. The model proposed for the chitinase was subjected to a dynamic analysis over a 1 ns interval, resulting in a model with 91.7% of the residues occupying favorable places on the Ramachandran plot and an RMS of 2.68.A enzima quitinase de Moniliophthora perniciosa, o fungo causador da doença vassoura de bruxa em Theobroma cacao, foi parcialmente purificada por precipitação com sulfato de amônio e filtração em coluna de Sephacryl S-200 usando fosfato de sódio como tampão de extração. A metodologia de Superfície de resposta (MSR) foi usada para determinar o pH e temperatura ótimos. Foram isoladas quatro quitinases diferentes: ChitMp I, ChitMp II, ChitMp III e ChitMp IV. A ChitMp I mostrou temperatura ótima de 44-73ºC e pH ótimo de 7,0-8,4. A ChitMp II apresentou temperatura ótima entre 45-73ºC e pH ótimo de 7,0-8,4. ChitMp III apresentou temperatura ótima de 54-67ºC e pH ótimo de 7,3-8,8. ChitMp IV apresentou temperatura ótima de 60ºC e pH ótimo em 7,0. Em relação à biologia computacional a sequência primária foi feita in silico usando banco de dados do Projeto Genoma/Proteoma do M. perniciosa, obtendo-se uma sequência com 564 pb e 188 aminoácidos, que foi utilizada para a obtenção da estrutura tridimensional por metodologia de modelagem comparativa. Os modelos gerados foram submetidos à validação usando Procheck 3.0 and ANOLEA. O modelo proposto para a quitinase foi submetido à análise dinâmica por 1 ns, resultando em um modelo com 91,7% de seus resíduos em locais regiões favoráveis no gráfico de Ramachandran e RMS de 2,68.

La presente investigación tuvo por objetivo caracterizar isoenzimáticamente a seis poblaciones de Annona cherimola de la Región La Libertad. Muestras de hojas tiernas de árboles con al menos dos fructificaciones fueron colectadas de las localidades de La Cuesta, Paday, Samne, Cascas, Poroto y Trujillo. Los perfiles electroforéticos de las enzimas PGI, APCH, MDH y ME fueron analizadas y procesadas. Encontramos polimorfismo sólo para los loci Pgi-1 y Me con siete y dos alelos respectivamente; valores promedio para polimorfismo de 0,40 y heterocigosidad media entre 0,206 (La Cuesta) y 0,285 (Poroto). Los resultados indican que existe una alta variabilidad fenotípica y genotípica en las seis poblaciones de Annona cherimola de la Región La Libertad.

La Colección Colombiana de Musáceas (CCM) es la única a nivel mundial que representa un alto valor por ser la que posee introducciones andinas de altura (> 1500 m.s.n.m.). La caracterización de este germoplasma puede generar valor agregado para su utilización en procesos de selección clonal y para el mejoramiento genético de la especie, mediante el uso de materiales diploides con características transmisibles de importancia. Por esta razón, 33 clones de la CCM conservadas in vitro, fueron evaluadas bioquímicamente mediante 10 enzimas, de las cuales cuatro fueron polimórficas: glutamato oxaloacetil transaminasa (GOT), ab-esterasa (ab-EST), peroxidasa (PRX) y diaforasa (DIAP). La enzima GOT fue la más discriminante entre grupos genómicos particulares. PRX, DIAP y ab-EST permitieron evaluar la variabilidad al interior de cada grupo. El estudio facilita el entendimiento de la estructura genética de los genotipos de plátano y banano cultivados en Colombia.The Colombian Collection of Musaceae (CCM) represents a great value to the world agriculture for being the only one with high Andean germplasm. The characterization of this germplasm can generate added value to use in processes of clone selection and breeding of the species by using diploid materials with desirable traits to be transferred and thus help alleviate food problems in poor countries. For this reason the CCM 33 clones conserved in vitro, were assessed biochemically using 10 enzymes of which four were polimorfics: oxaloacetil transaminasa glutamate (GOT), ab-esterase (ab-EST), peroxidase (PRX) and diaphorase (DIAP). The GOT enzyme was the most discriminating among individual genomic groups. PRX, ab-EST and DIAP allowed us to evaluate the variability within each group. The study provided a better understanding of the genetic structure of the genotypes of plantain and banana grown in Colombia.

INTRODUCTION: Isoenzymatic analyses were performed involving species of the Nyssorhynchus and Anopheles subgenera in order to estimate the intra and interspecies genetic variability. METHODS: Mosquitoes were caught at different localities in the Amazon region. The collection and rearing of mosquitoes in the laboratory followed specific protocols. For the genetic variability analyses, the technique of horizontal electrophoresis on starch and starch-agarose gel with appropriate buffer systems was used. The alloenzyme variation was estimated using the Biosys-1 software. RESULTS: Out of the 13 loci, eight were polymorphic. Anopheles nuneztovari presented the largest number of alleles per locus, while the smallest number was detected in Anopheles marajoara from Macapá. The largest number of polymorphic loci was found for Anopheles marajoara from Maruanum and the smallest for Anopheles benarrochi (Guayará Mirim). Anopheles darlingi (Macapá) presented the greatest heterozygosity (Ho = 0.167 ± 0.071), while the lowest heterozygosity (Ho = 0.045 ± 0.019) was observed in Anopheles intermedius (Pacoval) of the subgenus Anopheles. Wright's F coefficient revealed considerable genetic structuring between the populations of Anopheles darlingi (Fst = 0.110) and between the populations of Anopheles marajoara (Fst = 0.082). CONCLUSIONS: Considering all the species studied, the genetic distance ranged from 0.008 to 1.114. The greatest distance was between Anopheles mattogrossensis and Anopheles oswaldoi, while the smallest was between the Anopheles benarrochi populations.INTRODUÇÃO: Análises isoenzimáticas foram realizadas envolvendo espécies dos subgêneros Nyssorhynchus e Anopheles para estimar a variabilidade genética intra e interespecífica. MÉTODOS: Os mosquitos foram capturados em diferentes localidades da região Amazônica. A coleta e a criação dos mosquitos em laboratório foram conforme protocolos específicos. Na análise da variabilidade genética empregou-se a técnica de eletroforese horizontal em géis de amido e amido-agarose com sistemas tampões apropriados. A variação alozímica foi estimada pelo Programa Biosys-1. RESULTADOS: Dos 13 loci, oito foram polimórficos. Anopheles nuneztovari apresentou o maior número de alelos por loco, sendo o menor detectado para Anopheles albitarsis (Macapá). O maior número de loci polimórficos foi detectado em Anopheles marajoara (Maruanum) e o menor, em Anopheles benarrochi (Guayará Mirim). Anopheles darlingi (Macapá) apresentou a maior heterozigosidade (Ho = 0,167 ± 0.071) e a menor (Ho = 0,045 ± 0.019) foi observada em Anopheles intermedius (Pacoval) do subgênero Anopheles. O coeficiente F de Wright evidenciou considerável estruturação genética entre populações de Anopheles darlingi (Fst= 0,110) e entre as populações de Anopheles marajoara (Fst= 0,082). CONCLUSÕES: Considerando todas as espécies estudadas, a distância genética variou de 0,008 a 1,144, onde a maior distância foi entre Anopheles mattogrossensis e Anopheles oswaldoi e a menor, entre as populações de Anopheles benarrochi.

The internal morphology of embryos from immature and mature fruits of Heliconia bihai (L.) L. cv. Lobster Claw Two was examined. Embryos were inoculated into MS media (full MS and ½ MS) and GA3 (0, 2.5 and 5 mg L-1) with either sucrose or glucose. These plantlets were then replicated and transferred to MS medium (full MS or ½ MS) with 0 or 2.5 mg L-1 BAP and their multiplication was evaluated 30 and 45 days after inoculation. The genetic variability of the multiplied plants was estimated using isoenzyme analyses. The internal morphology of the mature embryos revealed their tissues to be in more advanced stages of differentiation than immature embryos. In the conversion phase, 85% of the inoculated embryos developed into plants in the ½ MS medium with sucrose, in contrast to only 41% of the embryos that were cultivated with glucose. In the multiplication phase, plants cultivated in ½ MS medium with 2.5 mg L-1 BAP demonstrated more buds. Isoenzyme analyses showed pattern changes in terms of the color intensity and the migration of some of the bands. These results may be associated with differences in the ages of the mother plants and of the plantlets obtained in vitro.Inicialmente os embriões procedentes de frutos imaturos e maduros de Heliconia bihai (L.) L. cv. Lobster Claw Two foram avaliados quanto à morfologia interna. Os embriões maduros foram inoculados em meio MS (1/2MS e MS completo) e GA3 (0, 2,5 e 5,0 mg L-1), utilizando sacarose ou glucose. Posteriormente, as plantas obtidas foram transferidas para meio 1/2MS acrescido de 0 ou 2.5 mg L-1 de BAP e aos 30 e 45 dias após a inoculação, avaliou-se a formação de gemas laterais. Aos 45 dias avaliou-se a variabilidade genética dessas plantas através da análise isoenzimática. Ao avaliar a morfologia interna observou-se que os embriões maduros apresentaram estádios de diferenciação superior aos embriões imaturos. Na fase de conversão do embrião em planta, observou-se que 85% dos embriões inoculados desenvolveram em plantas quando cultivados em meio ½MS com sacarose, contrastando com 41% cultivados no mesmo meio nutritivo acrescido de glucose. Na fase de multiplicação, as plantas cultivadas em meio ½MS acrescido de 2.5 mg L-1 de BAP apresentaram maior formação de gemas laterais. A análise isoenzimática apresentou diferença na intensidade de coloração e migração de algumas bandas. Esse comportamento pode estar associado com a diferença fisiológica relacionada à idade entre a planta mãe e as plantas provenientes do cultivo de embrião in vitro.

Spondias mombin L. is a fruit tree from the American continent from the Anacardiaceae family. In Brazil it is common in different vegetation types but is more frequent in the Atlantic and Amazonian rainforests. It is economically important because of its fruits, which are widely consumed raw or processed as fruit jellies, juices and ice creams. The leaves have great importance in the pharmaceutical industry because of their antibacterial properties. In the state of Pernambuco, cajá tree is widely distributed in the Zona da Mata region and less frequently in the Agreste and Sertão areas. In this work diversity and genetic structure were studied in four populations of cajá tree from Pernambuco's Zona da Mata, Northeast Brazil, using isozymes polymorphism analyses from electrophoreses. The result showed 100% of polymorphism (P) for nine alleles and the average of alleles per locus s was 2.4. The expected heterozygosity ranged from 0.530 to 0.574 and the observed heterozygosity , from 0.572 to 0.735. It was not observed inbreeding and the average F IT was -0.175, whereas within population inbreeding (f) varied from -0.08 to- 0.37. The genetic divergence among the populations (F ST) ranged from 0.006 to 0.028 and the average was 0.026. The average of estimated gene flow (Nm) was high (5.27). The CG-IPA population, corresponding to the germplasm collection of IPA, showed more than 96% of genetic similarity with other populations; therefore, it is a good representative of the existent genetic diversity in the Zona da Mata region.A espécie S. mombin L. é uma árvore frutífera de origem americana pertencente à família Anacardiaceae . No Brasil, é encontrada nos mais variados tipos de vegetação, porém é mais freqüente na Mata Atlântica e na Floresta Amazônica. Destaca-se economicamente por seus frutos serem amplamente consumidos in natura ou utilizados na indústria, principalmente para fabricação de polpas, geléias, sucos e sorvetes. As folhas têm grande importância para a indústria farmacêutica, por possuírem propriedades antibacterianas. No Estado de Pernambuco, a cajazeira encontra-se amplamente distribuída na Zona da Mata, ocorrendo com menos freqüência no Agreste e Sertão. Neste trabalho, estudaram-se a diversidade e a estrutura genética de populações de cajazeiras na Zona da Mata do Estado de Pernambuco, Nordeste do Brasil, por meio da análise do polimorfismo isoenzimático obtido por eletroforese. Os resultados revelaram 100 % de polimorfismo para nove alelos, sendo que o número médio de alelos por loco foi de 2,4. A heterozigosidade esperada variou de 0,530 a 0,574, e a heterozigosidade observada variou de 0,572 a 0,735. Não se verificou endogamia nas populações, sendo que o valor médio de F IT foi de -0,175, intrapopulacional ( f ) variou de -0,08 a -0,37. A diferenciação entre as populações (F ST) variou de 0,006 a 0,028, sendo que o valor médio de 0,026 e, o fluxo gênico estimado entre as populações (Nm) foram altos (5,27). A população CG-IPA, que constitui a Coleção de Germoplasma do IPA, apresentou acima de 96% de identidade genética com as demais populações e, portanto, tem alta representatividade na diversidade existente na região.

Se analizaron 16 sistemas isoenzimáticos para cinco especies colombianas del género Lutzomyia. Los niveles de heterocigosis media esperada insesgada oscilaron entre 0.098 (Lu. youngi) y 0.215 (Lu. torvida). Las cinco muestras estudiadas de forma global, para todos los marcadores analizados, presentaron desviación respecto al equilibrio Hardy-Weinberg por un exceso de homocigotos, tanto al utilizar algunas pruebas clásicas como tests exactos con cadenas de Markov. Este hecho puede estar favorecido por diversas causas: (1) la más probable es la existencia de efecto Wahlund en el seno de cada población debido a subdivisión y/o a la técnica de muestreo empleada. La endogamia puede descartarse ya que no todos los loci están afectados por el mismo tipo de exceso de homocigotos. (2) Sin embargo, no se puede descartar la existencia de alelos nulos, al menos, para algunos de los marcadores isoenzimáticos utilizados. El análisis jerarquizado con las F de Wright mostró valores elevados y significativos para cada uno de los estadísticos. El estadístico promedio F IT mostró un valor de 0.655 existiendo un conspicuo exceso de homocigotos a nivel total de todas las especies, el estadístico promedio F IS fue altamente positivo (0.515) mostrando exceso de homocigotos a nivel individual en cada una de las especies estudiadas. La heterogeneidad genética entre las cinco especies fue notable (F ST = 0.288). Esto muestra que esas especies están bien diferenciadas a nivel isoenzimático y que en el interior de cada especie también hay una subdivisión genética. La matriz de identidades genéticas de Nei muestra que las especies más relacionadas fueron Lu. longiflocosa-Lu. torvida (0.959) y Lu torvida-Lu. spinicrassa (0.960) mientras que las genéticamente más distantes fueron Lu. torvida-Lu. youngi (0.805) y Lu. quasitownsendi-Lu. youngi (0.796). Con los algoritmos UPGMA y Wagner, se observó que la especie más divergente fue Lu. youngi, mientras que las relaciones más conspicuas se observaron entre Lu. longiflocosa-Lu. torvida y Lu torvida-Lu. spinicrassa. Adicionalmente, con un análisis de autocorrelación espacial (índice de Moran) la mayoría de los alelos utilizados presentaron una estructura espacial muy débil o inexistente, lo que significa que los eventos de especiación entre las especies estudiadas se dieron en forma independiente de las distancias geográficas existentes actualmente entre ellas.Genetic structure in five Phlebotominae (Lutzomyia spp.), townsendi series, verrucarum group, in Colombia (Diptera: Prychodidae). Sixteen isoenzyme patterns were analyzed for five Colombian Lutzomyia species. The average unbiased expected heterozygosity levels ranged from 0.098 (Lu. youngi) to 0.215 (Lu. torvida). The five species samples, taken all the isoenzymes employed, were significantly deviated from the Hardy-Weinberg equilibrium by homozygous excess with classical as well as Markov chain exact tests. Possible causes: (1) Wahlund effect within populations due to subdivision and/or sampling. Endogamy could be discarded because these loci were affected by highly different levels of homozygous excess. (2) Null alleles could be not discarded, at least for some isoenzymes. The hierarchical Wright´s F analysis showed high and significant values for each parameter. The average F IT value was 0.655 with a conspicous homozygous excess at a global level (all species taken together); the average F IS value was significantly positive (0.515) as well, with homozygous excess within each species. The genetic heterogeneity between the fives species was noteworthy (F ST = 0.288), indicating clear genetic differentiation. The more related species pairs were Lu. longiflocosa-Lu. torvida (0.959) and Lu torvida-Lu. spinicrassa (0.960); while Lu. torvida-Lu. youngi (0.805) and Lu. quasitownsendi-Lu. youngi (0.796) were the most divergent (Nei´s genetic identity matrix). UPGMA and Wagner algorithms showed that the most divergent species was Lu. youngi, whereas the most related were Lu. longiflocosa-Lu. torvida and Lu torvida-Lu. spinicrassa. A spatial autocorrelation analysis (Moran´s I index) revealed a very weak, or inexistent spatial structure, which means that the speciation events between these species were independent from the geographic distances from where they currently live. Rev. Biol. Trop. 56 (4): 1717-1739. Epub 2008 December 12.

A cultura da aceroleira despertou um grande interesse do mercado consumidor, tendo em vista o alto teor de vitamina C (ácido ascórbico), que varia entre 1.325 a 2.250 mg por 100 mL de suco. Com a expansão da cultura, surgiram problemas fitossanitários, entre os quais a infecção das raízes da aceroleira por nematóides. Os produtos químicos utilizados no controle dos nematóides são agressivos ao meio ambiente, e a seleção de genótipos resistentes e tolerantes constitui-se na melhor alternativa para a solução do problema. O trabalho foi desenvolvido em casa de vegetação com 18 genótipos de aceroleira, com o objetivo de encontrar genótipos resistentes e tolerantes a Meloidogyne incognita raça 2 assistida por marcadores isoenzimáticos, para indicar plantas destinadas a porta-enxerto. A avaliação foi realizada 60 dias após a inoculação mediante análise das variáveis: número de ovos por planta e por grama de raiz, índice de galhas e massa de ovos, biomassa fresca relativa do sistema radicular e biomassa fresca relativa da parte aérea. Os resultados permitiram identificar o genótipo 023-CMF como menos suscetível, e os genótipos 027-CMF e 035-CMF, como mais suscetíveis. Estudos realizados através da eletroforese isoenzimática com os sistemas α esterase, fosfatase ácida e peroxidase, 20; 40 e 60 dias após a inoculação com ovos de M. incognita raça 2, possibilitaram relacionar a expressão de proteínas com a suscetibilidade. Os genótipos mais próximos geneticamente, com índice de similaridade 0,941, foram 027-CMF e 026-CMF, 046-CMF e 026-CMF, e 041-CMF e 026-CMF. O menor índice de similaridade genética (0,115) foi observado entre os genótipos 002-SPE e 036-CMF.The culture of the Barbados cherry has increased great interest in the consuming market, in view of high levels of vitamin C, each 100ml of juice has contents of 1,325mg / 2,250mg of vitamin C (Ascorbic acid).With the increase of the plantations phytosanitary problems appeared, such as infection of the roots for nematodes. The utilisation of chemical products for controlling nematodes is unsafe for the environment, therefore the election of resistant and tolerant genotypes is the best solution for this problem. The work was developed in greenhouse, with eighteen genotypes of Barbados cherry aiming to find resistant genotypes to Meloidogyne incognita race 2, marked with iso-enzymatic markers, indicating plants destined for rootstock. Sixty days after the inoculation, the evaluation of the variables began: amount of eggs per plant and weight of root, index of galls and eggs mass, fresh biomass relative to the root systems, fresh biomass relative to the aerial part. The results permitted to identify the genotype 023-CMF as the less susceptible and the genotypes 027-CMF and 035-CMF as the most susceptible. Studies carried out through iso-enzymatic electrophoresis with the systems α -esterase, acid phosphatase and peroxidase. Twenty, forty and sixty days after the inoculation with eggs of M. incognita race2, made it possible to relate the expression of proteins with the susceptibility. The genotypes genetically closer, with index of similarity 0.941 are: 027-CMF and 026-CMF, 046-CMF and 026-CMF and 041-CMF and 026-CMF. The lesser indices of genetic similarity 0.115 are: genotypes 002-SPE and 036-CMF.

La familia Lemnaceae carece de características morfológicas y anatómicas con fines comparativos y por ello se recurre a estudios moleculares para una diferenciación más precisa. En Costa Rica encontramos del género Lemna dos especies: Lemna aequinoctialis y Lemna valdiviana. Con el objetivo de una caracterización molecular y diferenciación de ambas especies se extrajeron isoenzimas esterasas. El estudio se llevó a cabo en el Laboratorio de Genética Molecular de la Escuela de Ciencias Biológicas de la Universidad Nacional, Heredia, Costa Rica. Se utilizó la técnica electroforesis en gel de poliacrilamida SDS-PAGE. Se encontró diferencia en el patrón de isoenzimas entre ambas especies. De acuerdo con previos estudios moleculares basados en la morfología y anatomía, flavonoides, isoenzimas y secuencias de ADN de ciertos genes, ambas especies son muy distintas; además pertenecen a grupos monofiléticos diferentes. Por su parte, la importancia de esta caracterización molecular radica en su posible uso como biorremediadoras, pues las esterasas han tenido un importante papel en la biorremediación. Asimismo, especies de la familia Lemnaceae son muy utilizadas en el tratamiento de aguas como biorremediadoras.

The aim of this study was to delimit two Iodophanus species: I. carneus and I. testaceus , based on morphological characteristics and electrophoretic patterns of their intracellular isozymes. Twenty monosporic strains were used, including five belonging to I. granulipolaris as a control. Fourteen isozyme systems were tested, and the five having the best resolution selected: aspartate aminotransferase, esterases, alkaline phosphatase, glutamate dehydrogenase, and superoxide dismutase. These analyses confirmed the similarity between I. carneus and I. testaceus , since they both produced the same band patterns, which were in turn different from the band pattern of I. granulipolaris. So, as we couldn´t find any character wich permit us to classify the isolated studied during this work in defferent species, we believe that I. testaceus shoul be consider as a synonym of I. carneus .El objetivo del presente trabajo fue la delimitación taxonómica de dos especies del género Iodophanus : I. carneus e I. testaceus a partir de caracteres morfológicos y de los patrones electroforéticos de isoenzimas intracelulares. Para ello se utilizaron veinte cepas monospóricas, cinco de las cuales pertenecientes a I. granulipolaris que fueron utilizadas como control. Se probaron catorce sistemas isoenzimáticos y se eligieron los cinco con mejor resolución: aspartato amino transferasa, esterasa, fosfatasa alcalina, glutamato dehidrogenasa y superóxido dismutasa. El análisis de los patrones isoenzimáticos corroboró la silimitud existente entre I. carneus e I. testaceus , ya que los patrones de bandas obtenidas para estas dos especies fueron iguales y diferentes de I. granulipolaris. Entonces, al no encontrar ningún caracter que nos permita separar a los aislamientos estudiados en este trabajo en dos especies distintas, proponemos a I. testaceus como un sinónimo de I. carneus.

Calophyllum brasiliense Camb. é uma espécie predominante de áreas com alta saturação hídrica, sendo comum nos ambientes cilares do sul de Minas Gerais. Com o intuito de se estudar a variabilidade genética dessa espécie, 20 indivíduos reprodutivos de C. brasiliense foram amostrados de duas populações localizadas a montante do Reservatório Hidroelétrico do Funil, localizado no Município de Lavras,MG. A análise de eletroforese isoenzimática permitiu a obtenção de 17 alelos, distribuídos entre oito locos, sendo estes representados em cinco sistemas enzimáticos: alfa-esterase, beta-esterase, fosfatase ácida, glutamato oxalacetato, malato desidrogenase e transaminase. Os índices de diversidade revelaram um baixo número de alelos por loco em ambas as populações (1,75 e 1,50). A porcentagem de locos polimórficos (P) foi de 37,5% e 50% nas populações I e II, respectivamente. A heterozigosidade média observada foi de 0,119 e 0,111 e a esperada, de 0,131 e 0,112. O número de migrantes (Nm) encontrado entre as populações foi de 2,70. O tamanho efetivo estimado foi de 18 indivíduos para a população I e 19 para a II.Calophyllum brasiliense Camb. is a predominant species in areas with high water saturation such as the riparian forests in southern Minas Gerais State. The objective of this research was to study the genetic variability of C. brasiliense populations. Twenty individuals were sampled from two populations located in Funil Dam in Lavras-MG. Isozyme eletrophoresis analysis provided evidence of 17 alleles distributed in 8 loci, which were represented in five enzymatic systems: alpha-esterase, beta esterase, acid phosphatase, malate dehydrogenase and transaminase oxalacetatum glutamate. The diversity indexes showed a low number of alleles per loci in both populations, pop I (1.75) and pop II (1.50). The polymorphic loci percentage (P) were 37.5% and 50% in populations I and II respectively. The mean heterozygosities were 0.119 and 0.111, while the expected was 0.131 and 0.112. The number of migrants (Nm) between populations was 2.70. The estimated effective size was 18 individuals for population I, and 19 for population II.

The objective of this work was to define the optimal conditions for invertase assay, seeking to determine the ideal parameters for the different isoenzymes of leaf and bark tissues in adult rubber trees. Assays of varying pH, sucrose concentration and temperature of the reaction medium were conducted for the two investigated isoenzymes. The results pointed out the existence of two different pH related isoforms for the two analyzed tissues, with an isoenzyme being more active at pH 5,5 and the other at neutral/alkaline pH. Leaf blade isoenzymes presented similar values for substrate concentration, whereas the bark isoenzyme presented maximum values below those previously reported. The assays at different temperatures presented similar values for leaf isoenzymes, though they have differed significantly among the obtained values.O objetivo deste trabalho foi definir as condições ótimas para a realização do ensaio enzimático da invertase, procurando-se determinar os parâmetros ideais para as diferentes isoenzimas de tecidos foliares e da casca de plantas adultas de seringueira. Foram realizados ensaios variando-se o pH, a concentração da sacarose e a temperatura do meio de reação para as duas isoenzimas estudadas. Os resultados indicaram a existência de duas isoformas diferentes em relação ao pH nos dois tecidos analisados, sendo uma isoenzima mais ativa a pH 5,5 e outra em pH neutro/alcalino. Com relação à concentração do substrato, as isoenzimas da lâmina foliar apresentaram valores semelhantes, enquanto a isoenzima da casca, valores máximos inferiores aos observados anteriormente. Os ensaios conduzidos em diferentes temperaturas tiveram valores semelhantes nas isoenzimas da folha, embora tenham diferido significativamente entre dos valores obtidos.

Some isozymes are involved in processes of interest to geneticists, such as flowering, plant development, height, pest and disease resistance, to name some. Therefore, it is important to determine, using biochemical markers, if there is differential expression during the vegetative and reproductive phases in the Aztec lily (Sprekelia formosissima (L.) Herbert). To do so, during those stages we collected tissue from the leaves, tepals gynoecium and stamens. These were then macerated, an extraction buffer was added and the total liquid extracts were used to determine isozymatic patterns via electrophoresis in starch gels. The differential enzymatic activity was evaluated using eleven isozymatic systems, from which only nine enzyme forms or isozymatic band patterns (IBP) were observed: two bands with peroxidase (POX; EC 1.11.1.7), three bands with acid phosphatase (ACP; EC 13.1.3.2), one with phosphoglucose isomerase (PGI; EC 5.3.1.9), two for phosphoglucomutase (PGM; EC 2.7.5.1) and one for malate dehydrogenase (MDH, EC 1.1.1.37), which showed the same pattern of isozymatic banding at all stages of the plant development. For the purpose of this research we only considered here the isoenzymatic banding patterns that were different. [ABSTRACT FROM AUTHOR]

The present study aimed to evaluate effects of different storage conditions on the castor bean seed cultivar IAC- 226 quality, stored for 12 months. For this purpose, seeds were stored in different environment and packaging conditions: in a cold chamber and conventional storage, using multiwall Kraft paper and in no vacuum and vacuum plastic packages at 1 atm; as well as under cryopreservation storage (-196 °C). Seed quality was evaluated before and after 4, 8 and 12 months of storage by germination tests, first count of germination, emergence percentage, emergence speed index and determination of changes in catalase (CAT) and superoxide dismutase (SOD) enzyme systems. Cryopreservation (-196 °C) is efficient in maintaining the physiological quality of castor bean cultivar IAC-226 for 12 months. The enzyme catalase stands out as a marker of castor seed deterioration during storage. [ABSTRACT FROM AUTHOR]

The action of the metalloenzyme carbonic anhydrase (AC) in plants has been mainly associated to increase the CO2 concentration inside chloroplast to enhance the carboxylation rate of Rubisco enzyme, being this reaction that integrates the CO2 in carbohydrates during photosynthesis. However, the AC activity has been found in non-green tissues also, like roots, nodules, etiolated leaves, seeds, as well as in bacteria and fungi, which plays diverse and specific rolls. In the present study, the isoenzyme patterns of AC were analyzed in tomato (Solanum lycopersicum L.) roots inoculated in individual way with six different arbuscular mycorrhizal fungi (AMF) strains, taking the samples at two moments of symbiosis, and afterward they were compared among them. Results revealed that the AM fungus presence provoked the appearance of isoenzymes with AC activity in tomato roots cv. 'AMALIA', and the obtained isoenzymatic patterns showed a differential response dependent on inoculated strain, principally at early stages of symbiosis. The possible fungal origin of observed isoenzymes is discussed. The present work constitutes the first report about AC isoenzymes in AMF-colonized roots. [ABSTRACT FROM AUTHOR]

Soil and root samples were collected in soybean from the North, Northwest and South regions of the Rio Grande do Sul. Plant-parasitic nematodes were identified from 40 samples, and the species of rootknot nematode (Meloidogyne spp.) and spiral nematode (Helicotylenchus spp.) were determined by both esterase phenotype profile and morphological and morphometric characters. Nematodes detected in the soil samples included Meloidogyne (65%), Helicotylenchus (100%), Tylenchus (72.5%), Aphelenchus (17.5%), Paratylenchus (15%), and Pratylenchus (10%). Species of root-knot nematode identified in root samples included predominantly M. javanica Est J3 (82%), followed by M. arenaria Est A2 (31%), and M. morocciensis (6%). Species of spiral nematodes that were detected included H. dihystera (78%), H. pseudorobustus (11%), and H. multicinctus (11%) in the soil. [ABSTRACT FROM AUTHOR]

Bromus auleticus é uma espécie perene, nativa do estado do Rio Grande do Sul, que tem demonstrado adaptabilidade, potencial para produção de forragem de qualidade e alta diversidade morfológica. Neste trabalho, foram analisados padrões de bandas de sistemas enzimáticos e de marcadores RAPD, com o objetivo de determinar a variabilidade genética em 16 acessos de Bromus auleticus do Rio Grande do Sul. A variabilidade foi avaliada pelo índice de similaridade de Jaccard, utilizando-se o método UPGMA nas análises de agrupamento. Os índices de similaridade variaram de 0,0 a 0, 50 com base em isoenzimas e de 0,15 a 0,71 com base em marcadores RAPD. Os dados foram eficientes para a formação de grupos, indicando a variabilidade genética dos acessos analisados. Entretanto, houve baixa relação entre os agrupamentos e os locais de coleta. A variabilidade genética dos acessos é importante sob o ponto de vista do melhoramento genético, permitindo a seleção futura dos genótipos a partir de seus respectivos desempenhos. A utilização simultânea de isoenzimas e RAPD foi eficiente na caracterização da diversidade genética dos acessos analisados.Bromus auleticus is a perennial, native species of the state of Rio Grande do Sul, which has demonstrated adaptability, potential yield of good quality forage as well as high morphological diversity. This work has analyzed band patterns from enzymatic systems and RAPD markers, with the objective to access the genetic variability in 16 accessions of Bromus auleticus from Rio Grande do Sul. The variability was evaluated using Jaccard similarity index. The method of grouping based on the average (UPGMA) was used in cluster analyses. The similarity index ranged from 0.0 to 0.50 using isozumes and from 0.15 to 0.71 using RAPD markers. The data have been efficient for the formation of different groups, indicating the genetic variability of the accession analyzed. However, these groupings have little relationship with the respective collecting places. The genetic variability of the accessions is important to the genetic improvement, allowing future genotype selection based on their respective performances. The simultaneous utilization of isozymes and RAPD was efficient in characterizing the genetic diversity of the accessions evaluated.

O objetivo deste trabalho foi estudar, por locos isoenzimáticos, a diversidade e a estrutura genética espacial de genótipos de Myracrodruon urundeuva em duas populações naturais, uma no Sudoeste (Selvíria-SEL) e outra no Sudeste (Paulo de Faria-PFA) do Brasil. Para isso, foram avaliados cinco sistemas isoenzimáticos, 25 e 30 indivíduos adultos das populações SEL e PFA, respectivamente. A estimativa da divergência genética entre populações foi baixa (=0,043). As heterozigosidades observada e esperada foram altas nas populações (0,317 e 0,511, respectivamente), e o excesso significativo de heterozigotos foi detectado na população PFA (= -0,252). A análise da distribuição genética espacial dos genótipos a partir do índice I de Moran revelou estruturação significativa até 5.224 m na população mais explorada (SEL, =0,09) e tendência à distribuição aleatória na população menos explorada (PFA, = -0,02). A provável causa da estruturação na população SEL foi a dispersão de sementes próxima às árvores-matriz, associada ao processo de recolonização a partir de sementes oriundas de poucos genótipos remanescentes. As implicações dos resultados são discutidas do ponto de vista da conservação e do melhoramento genético.The diversity and spatial genetic distribution of Myracrodruon urundeuva genotypes were studied in two Brazilian populations in the Southwest (Selvíria-SEL) and Southeast Brazilian regions (Paulo de Faria-PFA). Twenty-five and thirty adult individuals were evaluated for five allozyme systems in SEL and PFA populations, respectively. Estimates of the genetic divergence between populations were low (=0.043). Observed and expected heterozygosities were high in both populations (0.317 to 0.511, respectively). Significant and excessive number of heterozygotes was detected in PFA population (=-0.252). The spatial distribution analysis through Moran's index I revealed a significant structuring up to 5,224 m in the more exploited population (SEL, =0.09) and a trend to randomness in the less exploited area (PFA, = -0,02). Seed dispersion near mother trees and the recolonization process through seeds from few genotypes are probable causes of the structuring in the SEL population. The implications of the results are discussed from the conservation and breeding point of views.

A herança e o desequilíbrio de ligaçãoforam estudadas em seis locos polimórficos (Got-1,Got-2, Lap-1, 6pgdh-1, Pgi-2 e Pgm-1) de cincosistemas isoenzimáticos (GOT, LAP, PGI, 6PGD ePGM) revelados em 55 progênies de polinizaçãoaberta de duas populações de Myracrodruonurundeuva Fr. All. (aroeira) do Estado de SãoPaulo. A relação de ligação entre locos foi estudadapela medida composta de desequilíbrio de ligaçãode Burrows. A segregação para os locos foigeralmente homogênea entre árvores. No loco Got-1foi detectado desvio da razão de segregação esperada1:1. Evidências de desequilíbrio de ligação foramobservadas entre os locos: Lap-1:Pgi-2.Inheritance and linkage disequilibriumwere studied in six polymorphic enzyme loci(Got-1, Got-2, Lap-1, 6pgdh-1, Pgi-2 and Pgm-1)from five isozymes systems (GOT, LAP, PGI,6PGD e PGM) in 55 open-pollinated families oftwo Myracrodruon urundeuva Fr. All. (aroeira)populations of São Paulo State. Burrowscomposite measure of linkage disequilibrium wasused to study linkage relationship between loci.Gene segregation of loci was generallyhomogeneous among trees. Got-1 locus showedsignificant departure from the expected 1:1segregation ratio. Linkage disequilibrium evidencewas detected between a pair of loci: Lap-1:Pgi-2.

A herança e o desequilíbrio de ligaçãoentre 11 locos polimórficos de oito sistemasisoenzimáticos foram estudados em progênies depolinização aberta de duas populações deEschweilera ovata (Bambess.) Miers (biriba) doEstado da Bahia. A análise da segregação deprogênies de árvores heterozigotas revelou simplesherança mendeliana para onze locos (Acp-1, Dia-1,Dia-2, Dia-3, G6pdh-1, Mdh-1, Nadh-1, Nadh-2,Pgi-1, Pgi-2 e 6pgdh-1). A segregação desseslocos foi geralmente homogênea entre árvores,embora algumas tenham apresentado desviossignificativos da segregação esperada 1:1.Desvios significativos do equilíbrio gaméticoforam detectados entre três pares de locos:6pgdh-1:Pgi-2, Dia-1:Dia-2 e Dia-2:Dia-3.Inheritance and gametic disequilibriumwere studied between 11 polymorphic loci of theeight isozymes systems from open-pollinatedfamilies of two Eschweilera ovata (Bambess.)Miers (biriba) populations from Bahia State.Segregation analysis of families from heterozygotetrees revealed simple mendelian inheritance foreleven loci (Acp-1, Dia-1, Dia-2, Dia-3, G6pdh-1,Mdh-1, Nadh-1, Nadh-2, Pgi-1, Pgi-2 and 6pgdh-1).Gene segregation at these loci was generallyhomogeneous among trees. However, there weresignificant deviations from the expectedsegregations ratio 1:1 at some individual trees.Significant gametic disequilibrium was observedfor three pairs of loci: 6pgdh-1:Pgi-2, Dia-1:Dia-2and Dia-2:Dia-3.