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Sample records for al azar rapd

  1. Low Genetic Diversity Among Garlic (Allium sativum L. Accessions Detected Using Random Amplified Polymorphic DNA (RAPD Escasa Diversidad Genética entre Accesiones de Ajo (Allium sativum L. Detectada Mediante ADN Polimórfico Amplificado al Azar (RAPD

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    Mario Paredes C

    2008-03-01

    Full Text Available Garlic (Allium sativum L. is a species of vegetative propagation, showing high morphological diversity. Besides, its clones have specific adaptations to different agroclimatic regions. The objective of this study was to determine the genetic diversity of 65 garlic clones collected in Chile and introduced from different countries, by using RAPD (Random Amplified Polymorphic DNA. Fourty random primers of 10 mers generated a total of 398 bands with an 87% of polymorphism. Each primer amplified between two and 20 bands. The size of the fragments obtained fluctuated between 3200 and 369 bp. The results showed that the clones analyzed had a genetic similarity rate of 94%. In addition, 70% of them were clustered in one major group. However, in spite of that situation several clones have different agronomic characteristicsEl ajo (Allium sativum L. es una especie de propagación vegetativa, que presenta una amplia variabilidad morfológica. Los clones de esta especie tienen una adaptación específica a diferentes regiones agroclimáticas. El objetivo de este estudio fue determinar la diversidad genética existente en 65 clones de ajos colectados en Chile e introducidos desde diferentes países, utilizando RAPD (ADN Polimórfico Amplificado al Azar. Para esta evaluación se utilizaron 40 partidores de 10-mers. Los partidores generaron entre dos y 20 bandas, observándose un alto número de patrones con bandas múltiples. Los fragmentos generados difieren en su tamaño entre 3.200 y 369 pb. Los partidores generaron 398 bandas, de las cuales un 87% fueron polimórficas. El análisis estadístico realizado detectó una similitud genética alta, de un 94% entre las accesiones evaluadas, donde aproximadamente un 70% de los clones formaron un grupo homogéneo. Sin embargo, este grupo incluye clones que presentan diferentes características agronómicas

  2. Diversidad genética entre especies del género lemna (lemnaceae utilizando fragmentos polimórficos de ADN amplificados al azar (rapds

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    Frank Solano Campos

    2016-03-01

    Full Text Available Lemna aequinoctialis and Lemna valdiviana were tested genetic similarity by random amplified polymorphic DNA (RAPD fingerprinting. Of 60 primers (10-mer tested, 26 generated polymorphic products. 143 bands were found from 200bp up to 1500bp, 109 were polymorphic (76,2%, with an average of 4,19. Of these, only 18 were specific of L. aequinoctialis, while 91 bands were specific for L. valdiviana. Data were used to generate Jaccard’s similarity coefficients and to construct a dendrogram using UPGMA method in the BiodiversityPro statistical package. It is concluded from this study that there were clear differences (high genetic diversity between L. aequinoctialis and L. valdiviana and that RAPD sesults are comparable with those obtained from studies on morphology, It is a practical method to assess the relationships between these species.

  3. Optimization of random amplified polymorphic DNA techniques for use in genetic studies of Cuban triatominae Optimización de la técnica de ADN polimórfico amplificado al azar (RAPD para su utilización en la caracterización genética de triatomíneos cubanos

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    Jorge Fraga

    2005-10-01

    Full Text Available Random amplified polymorphic DNA (RAPD technique is a simple and reliable method to detect DNA polymorphism. Several factors can affect the amplification profiles, thereby causing false bands and non-reproducibility of assay. In this study, we analyzed the effect of changing the concentration of primer, magnesium chloride, template DNA and Taq DNA polymerase with the objective of determining their optimum concentration for the standardization of RAPD technique for genetic studies of Cuban Triatominae. Reproducible amplification patterns were obtained using 5 pmoL of primer, 2.5 mM of MgCl2, 25 ng of template DNA and 2 U of Taq DNA polymerase in 25 µL of the reaction. A panel of five random primers was used to evaluate the genetic variability of T. flavida. Three of these (OPA-1, OPA-2 and OPA-4 generated reproducible and distinguishable fingerprinting patterns of Triatominae. Numerical analysis of 52 RAPD amplified bands generated for all five primers was carried out with unweighted pair group method analysis (UPGMA. Jaccard's Similarity Coefficient data were used to construct a dendrogram. Two groups could be distinguished by RAPD data and these groups coincided with geographic origin, i.e. the populations captured in areas from east and west of Guanahacabibes, Pinar del Río. T. flavida present low interpopulation variability that could result in greater susceptibility to pesticides in control programs. The RAPD protocol and the selected primers are useful for molecular characterization of Cuban Triatominae.La técnica de ADN polimórfico amplificado al azar (RAPD es un método simple para detectar el polimorfismo genético del ADN. Diferentes factores afectan los perfiles de amplificación lo que se manifiesta en la presencia de bandas falsas y en la reproducibilidad del ensayo. En nuestro trabajo analizamos los cambios de la concentración de cebador, ADN molde, cloruro de magnesio y de Taq ADN polimerasa con el objetivo de determinar su

  4. Contribución al estudio epidemiológico del kala-azar en colombia

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    Corredor Arjona, Augusto; Osorno Mesa, Ernesto; Parra Márquez, Jorge

    2011-01-01

    Los autores informan sobre el estudio epidemiológico del kala-azar en un foco situado en el municipio de Melgar, departamento del Tolima. Colombia. Encontraron la modalidad del kala-azar de la zona neotropical descrita en otras partes de América; mayor incidencia en los grupos humanos menores de cinco años, perros frecuentemente infectados, presencia de lutzomyia longipalpis. Hacen una discursión sobre la aparición de nuevos focos de kala-azar relacionándolos con la suspensión del rociado de ...

  5. Microsatélites amplificados al azar (RAM en estudios de diversidad genética vegetal

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    Jaime Eduardo Muñoz Flórez

    2008-12-01

    Full Text Available Se revisó el uso e importancia, ventajas, desventajas y características de la técnica Microsatélites Amplificados al Azar (RAM en uchuva Physalis peruviana, mora Rubus spp, guayaba Psidium guajava y heliconias Heliconia spp. En mora se diferenciaron las especies R. glaucus, R. robustus y R. urticifolius, se detectaron duplicados y se encontró alta variabilidad genética en R. glaucus, la especie más importante. En uchuva se encontró alta diversidad y dos accesiones de fruto rojo que se diferenciaron genéticamente de las amarillas y una región geográfica con alta variabilidad. En guayaba los cebadores fueron altamente polimórficos y se encontró alta variabilidad en el Valle del Cauca. En heliconias y especies relacionadas se diferenciaron las familias del orden Zingiberales, algunos subgéneros y variaciones en la especie. La técnica es de bajo costo, utiliza un cebador, no requiere información previa, es altamente polimórfica y diferencia especies en los taxones evaluados.

  6. Microsatélites amplificados al azar (RAM en estudios de diversidad genética vegetal

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    Muñoz Flórez Jaime Eduardo

    2008-12-01

    Full Text Available Se revisó el uso e importancia, ventajas, desventajas y características de la técnica Microsatélites Amplificados al Azar (RAM en uchuva Physalis peruviana, mora Rubus spp, guayaba Psidium guajava y heliconias Heliconia spp. En mora se diferenciaron las especies R. glaucus, R. robustus y R. urticifolius, se detectaron duplicados y se encontró alta variabilidad genética en R. glaucus, la especie más importante. En uchuva se encontró alta diversidad y dos accesiones de fruto rojo que se diferenciaron genéticamente de las amarillas y una región geográfica con alta variabilidad. En guayaba los cebadores fueron altamente polimórficos y se encontró alta variabilidad en el Valle del Cauca. En heliconias y especies relacionadas se diferenciaron las familias del orden Zingiberales, algunos subgéneros y variaciones en la especie. La técnica es de bajo costo, utiliza un cebador, no requiere información previa, es altamente polimórfica y diferencia especies en los taxones evaluados.

  7. Caracterización molecular con microsatélites amplificados al azar (RAMs de Inchi (Caryodendron orinocense K.

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    Ana Cruz Morillo Coronado

    2015-05-01

    Full Text Available Título en ingles: Molecular characterization with random amplified microsatellites (RAMs of Inchi (Caryodendron orinocense K.Resumen: El Inchi o Cacay (Caryodendron orinocense Karsten es una de las especies más promisorias de la Amazonía y la Orinoquia colombiana.  El principal producto del Cacay son sus almendras, de las que se extrae un aceite con aplicaciones cosméticas, fitoterapéuticas y alimenticias, además presenta un alto contenido de antioxidantes como los Omega 3, 6 y 9 y Vitaminas como la A y E. No existen estudios sobre la caracterización molecular de este recurso fitogenético, por lo cual el objetivo de esta investigación fue caracterizar la diversidad genética usando marcadores Microsatélites Amplificados al Azar (RAMs. El análisis de similitud al 0.50 formó cuatro grupos de acuerdo al sitio geográfico, siendo los materiales procedentes de Putumayo, Cacayal 19, Pauna y Castilla los de menor similitud. Los valores de heterocigosidad estimada fueron de 0.16 y 0.28 para los cebadores CGA y GT, respectivamente. El porcentaje de loci polimórfico varió entre 55% para el cebador CGA y el 90% para el GT. El valor de Fst promedio para los 27 materiales estudiados fue de 0.35, mostrando que la dinámica espacio-temporal de los materiales de Caryodendron tienden hacia una diferenciación genética, propio de sus procesos evolutivos e incidencia de la domesticación.Palabras clave: cacay, marcadores moleculares, diversidad genética, flujo genético, domesticación.Abstract: The Inchi or Cacay (Caryodendron orinocense Karsten is one of the most promising species of the Amazon and Orinoco Colombian. The main product of Cacay are its almonds, from extracted oil cosmetic, phytotherapeutic and food applications, also has a high content of antioxidants such as Omega 3, 6 and 9 and vitamins like A and E. There are no studies on the molecular characterization of this plant genetic resource; therefore the objective of this research

  8. Azar y Aritmetica

    CERN Document Server

    Helfgott, Harald Andres

    2009-01-01

    Let omega(n) be the number of prime divisors of an integer n. Let n be an integer taken at random between 1 and N. What can be said about the value then taken by omega(n)? What is its expected value? What is its distribution in the limit? What is the probability that omega(n) will deviate greatly from its expected value? We will study these questions as an introduction to probabilistic number theory. We treat several central topics in probabilistic number theory without assuming previous knowledge of the area. Neither measure theory nor complex analysis are assumed. In the exercises, among other topics, we develop some of the bases of sieve theory as an application of probabilistic ideas. ----- Sea omega(n) el numero de divisores primos de un entero n. Sea n un entero tomado al azar entre 1 y N. Que se puede decir del valor que entonces tomara' omega(n)? Cual es su esperanza? Cual es su distribucion en el limite? Cual es la probabilidad que omega(n) tome valores que se alejen mucho de su esperanza? Estudiamos...

  9. Microsatélites amplificados al azar (RAM en estudios de diversidad genética vegetal Random amplified microsatellites (RAM´s in plant genetic diversity studies

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    Jaime Eduardo Muñoz Flórez

    2008-12-01

    Full Text Available Se revisó el uso e importancia, ventajas, desventajas y características de la técnica Microsatélites Amplificados al Azar (RAM en uchuva Physalis peruviana, mora Rubus spp, guayaba Psidium guajava y heliconias Heliconia spp. En mora se diferenciaron las especies R. glaucus, R. robustus y R. urticifolius, se detectaron duplicados y se encontró alta variabilidad genética en R. glaucus, la especie más importante. En uchuva se encontró alta diversidad y dos accesiones de fruto rojo que se diferenciaron genéticamente de las amarillas y una región geográfica con alta variabilidad. En guayaba los cebadores fueron altamente polimórficos y se encontró alta variabilidad en el Valle del Cauca. En heliconias y especies relacionadas se diferenciaron las familias del orden Zingiberales, algunos subgéneros y variaciones en la especie. La técnica es de bajo costo, utiliza un cebador, no requiere información previa, es altamente polimórfica y diferencia especies en los taxones evaluados.The use and importance, advantages, disadvantages and features of the Random Amplified Microsatellites RAMs technique, were reviewed in Cape gooseberry Physalis peruviana, blackberry Rubus spp, guava Psidium guajava and heliconias Heliconia spp. In blackberry, we differentiated the species R. glaucus, R. robustus y R. urticifolius, detected duplicated accessions and found high genetic diversity in R. glaucus, the most important specie. In cape gooseberry we found high diversity and two red fruit accessions genetically differentiated from the yellow fruit ones and a geographical region with high variability. In guava, primers were highly polymorphic and found high variability in Valle del Cauca region. In Heliconia and related species we differentiated families belonging to Zingiberal order, between some sub genera and variation among specie. The technique has low cost of implementation, use a single primer, do not require previous information, is highly

  10. Evolution of the histones: free play with exon shuffling Evolucion de las histonas: Juego libre con reordenamiento de exones al azar

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    G. CECILIA TORO

    2001-03-01

    cromatina es el nucleosoma, formado por 168 pares de bases de DNA enrollados en el octámero de histonas, el cual consiste en dos copias de las histonas H2A, H2B, H3 y H4. El DNA es sellado en los puntos de entrada y salida por una histona de unión: la histona H1. Se supone que las histonas son proteínas muy conservadas. Sin embargo, durante los últimos años se ha encontrado que estas proteínas presentan un alto grado de divergencia en varios eucariontes inferiores. En Trypanosoma se ha encontrado que las histonas H3 y H4, que están en el centro de la organización nucleosomal, muestran más de un 30 % de divergencia, y que la histona H1 corresponde sólo a uno de los tres dominios presentes en los eucariontes superiores. Estos rasgos en Trypanosoma podrían explicar, al menos en parte, la ausencia de condensación de la cromatina en cromosomas durante la división celular en estos parásitos. La evolución de las histonas ha sido considerada como algo peculiar, con varias propuestas difíciles de reconciliar con los datos experimentales. En este trabajo se propone que las histonas han seguido la misma historia evolutiva de muchas otras proteínas. Si consideramos que los exones codifican para dominios estructurales y funcionales en las proteínas y que, en el origen de los eucariontes, las histonas, al igual que otras proteínas, podrían haberse formado por unidades (teoría tipo mecano, se podría esperar que estas unidades o dominios eventualmente fuesen encontrados en organismos actuales que exhiban características primitivas. Mas aún, estas unidades podrían funcionar independientemente. Nuestros resultados relativos a la estructura de los genes de histonas y de sus proteínas en Trypanosoma cruzi, y el análisis de otras histonas de diferentes especies confirman la proposición

  11. Juegos de azar: aleatoriedad y razonamiento falaz

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    Iranzo, Valeriano

    2012-01-01

    El artículo consta de cuatro apartados. El primero hace un breve recorrido histórico para mostrar la estrecha imbricación entre los juegos de azar y las matemáticas. En el segundo se argumenta que los juegos de azar son 'injustos' para el apostante y favorables para la casa de apuestas; precisamente en eso reside el margen de benefi cios que obtiene esta última y que la convierte en un negocio rentable. Se explica además cómo, mediante un sencillo cálculo, podemos averiguar si ...

  12. Caracterización de tilapia roja (Oreochromis sp. con marcadores moleculares RAPD Characterization of the red tilapia Oreochromis sp. through molecular markers RAPD

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    Julieta Torres Jaramillo

    2010-04-01

    Full Text Available Se utilizó la técnica RAPD (amplificación al azar de ADN polimórfico para el estudio de la diversidad genética de Oreochromis sp. (tilapia roja en cinco piscícolas del Valle del Cauca (Colombia y en la determinación del nivel de introgresión de las especies parentales Oreochromis mosambicus, O. niloticus y O. aureus. Se evaluaron 25 cebadores, ocho fueron polimórficos y se obtuvieron 109 bandas. Los valores de heterocigosidad esperada (0.196 a 0.256 y la estructura genética (Gst = 0.22 para Oreochromis sp. indicaron un elevado grado de polimorfismo y alta estructuración genética. Estos resultados fueron consistente con el Fst = 0.268 (P Random amplified polymorphic DNA (RAPD markers were used to study genetic diversity on red Tilapia (Oreochromis sp. species collected from five fish farms located in the Valle del Cauca, Colombia and to determine the level of introgression from three parental species O. mosambicus, O. niloticus and O. aureus into local Oreochromis populations. from the 25 RAPD primers evaluated, eight were polymorphic and 109 banding patterns were observed, any of them were specific. The expected levels of heterozygosis (0.1964 to 0.2561 and genetic structure (Gst = 0.22 funded for Oreochrosmis sp. indicate high grade of polymorphism and genetic structuring. This results were observed following the analysis of molecular variance [AMOVA] (Fst = 0.268 (P <0.0001 and Multiple correspondence analysis (Gst = 0.040. The values of genetic similarity, the analysis of group, the analysis of multiple correspondence and the level of introgression, indicated that the differences in the introgression levels(P=0.0001 were significant. The low level of observed genetic differentiation among populations, could be the result of fish with the same genetic origin, whereas the high variation within populations can be displayed by handling practices and the pressure of selection to favor commercial phenotypes. The level of introgression

  13. Genetic Variability of the Tomato Leaf Miner (Tuta absoluta Meirick; Lepidoptera: Gelechiidae, in Tunisia, Inferred from RAPD-PCR Variabilidad Genética del Minador de Hojas de Tomate (Tuta absoluta Meyrick; Lepidoptera: Gelechiidae en Túnez desde RAPD-PCR

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    Asma Bettaibi

    2012-06-01

    Full Text Available The tomato leaf miner Tuta absoluta Meyrick has invaded tomato (Solanum lycopersicum L. crop in Tunisia since 2008 and is representing today a major threat to the production of this crop. In this study, we used the Randomly Amplified Polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR technology to assess the genetic variability within and among seven populations of T. absoluta, collected on tomato from different regions in Tunisia. Using five RAPD-PCR primers and 108 individuals, 140 polymorphic fragments were recorded. From 335 different RAPD phenotypes generated, 71 were redundant and 264 unique to a specific population. The genetic structure of T. absoluta was investigated using analysis of molecular variance (AMOVA, genetic distances (Fst and multidimensional scaling (MDS. We detected a high genetic diversity within and among populations in conjunction with a significant differentiation between populations, suggesting that different founder genotypes would have been responsible of the introduction of T. absoluta in Tunisia. The presence of overlapping phenotypes probably indicates migration events between populations, mainly through infested plant material carried by humans.El minador de hojas de tomate Tuta absoluta Meyrick ha invadido el cultivo del tomate (Solanum lycopersicum L. en Túnez desde 2008 y actualmente representa una importante amenaza para su producción. En este estudio usamos la tecnología de ADN polimórfico amplificado al azar-reacción de cadena polimerasa (RAPD-PCR para evaluar la variabilidad genética dentro y entre siete poblaciones de T. absoluta, colectadas desde tomate en diferentes regiones de Túnez. Usando cinco primers RAPD-PCR y 108 individuos, se registraron 140 fragmentos polimórficos. Se generaron 335 fenotipos RAPD diferentes, entre los cuales 71 fueron redundantes y 264 únicos para una población específica. La estructura genética de T. absoluta se investigó usando análisis de varianza molecular

  14. Jaundice in pediatric visceral leishmaniasis (kala-azar)pa-tients

    Institute of Scientific and Technical Information of China (English)

    AKMMamunur Rashid; MdAsrafuzzaman; Abdullah Al Mamun; Abdullah Al Mahboob

    2009-01-01

    Objective:Visceral leishmaniasis (Kala-azar)is endemic in many countries including Bangladesh.Clinical presentation of visceral leishmaniasis in children and adult may vary and at time may simulate many tropical and hepatobiliary diseases.Jaundice and ascites are not common in kala-azar patients.Methods:During the period of January 2005 to December 2006,all the records of the confirmed kala-azar patients presented with jaundice were included in this study.Kala-azar was confirmed by serology test ICT (Immuno Chromatography) and Bone Marrow study.Results:Total 12 kala-azar patients were encountered during this period.Among these twelve cases,presenting features were jaundice (7),splenomegaly (12),hepatomegaly (11)and asci-tes (4).Initial clinical diagnosis of chronic liver disease (CLD)was made in (5),Congenital hemolytic a-naemia in (1)and kala-azar in rest of the patients (6).Common leucopenia and relative lymphocytosis was not observed in any patients.Conclusion:Kala-azar may present with various clinical manifestation in chil-dren and adult.Jaundice can be considered to be a common manifestation particularly in pediatric kala-azar patients.Otherwise,it may mislead to another diagnosis if it is taken as a rare feature in kala-azar.

  15. Estudio del polimorfismo genético de las células de la médula ósea y del sistema nervioso central de ratas mediante la técnica de RAPD

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    Esteban Alberti

    2011-07-01

    Full Text Available Study of genetic polymorphism of the bone marrow and the central nervous system of rats cells by RAPD technique  Título corto: RAPD a células de ratas. ResumenLos modelos experimentales en rata han sido de gran utilidad en las evaluaciones terapéuticas o de reemplazo de células en enfermedades neurodegenerativas. Se ha comprobado que las células de la médula ósea (CMO de ratas pueden diferenciarse en células que no forman parte de sus linajes normales. Hay evidencias de estos procesos de trans-diferenciación, pero aún no se conocen los mecanismos moleculares que activan estos procesos. El propósito de nuestro trabajo fue estudiar el polimorfismo genético del ADN de los tipos celulares que conforman las CMO y las células del sistema nervioso central (SNC, estríatales y de la corteza de ratas mediante la técnica de RAPD. Las CMO, las células mononucleares (CMMO, las células estromales (CEMO y las del SNC fueron obtenidas de ratas, y su ADN genómico fue purificado y amplificado mediante la técnica de RAPD, utilizando 15 cebadores al azar. Se construyó un dendograma de las bandas de amplificación generadas utilizando el método de UPGMA. Las células estudiadas según el análisis del RAPD quedaron en 2 grupos bien definidos, pudiéndose diferenciar las CEMO del resto de las células estudiadas. Los cebadores OPA-6, 7 y 12, mostraron el polimorfismo genético de los linajes de células estudiadas. Mediante la técnica de RAPD se demostró la variabilidad genética entre las CEMO y las CMMO, células estriadas y de corteza que mostraron una homogeneidad genética, proponiéndose marcadores específicos de RAPD para cada grupo de células. Este es el primer estudio del polimorfismo genético de las CMO y del SNC de ratas.  Palabras clave: células de la médula ósea (CMO, células del sistema nervioso central, polimorfismo genético, RAPD. AbstractExperimental models have been of grate usefulness in the therapeutic or

  16. Caracterización de tilapia roja (Oreochromis sp. con marcadores moleculares RAPD

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    Julieta Torres Jaramillo

    2010-04-01

    Full Text Available Se utilizó la técnica RAPD (amplificación al azar de ADN polimórfico para el estudio de la diversidad genética de Oreochromis sp. (tilapia roja en cinco piscícolas del Valle del Cauca (Colombia y en la determinación del nivel de introgresión de las especies parentales Oreochromis mosambicus, O. niloticus y O. aureus. Se evaluaron 25 cebadores, ocho fueron polimórficos y se obtuvieron 109 bandas. Los valores de heterocigosidad esperada (0.196 a 0.256 y la estructura genética (Gst = 0.22 para Oreochromis sp. indicaron un elevado grado de polimorfismo y alta estructuración genética. Estos resultados fueron consistente con el Fst = 0.268 (P < 0.0001 dado por el Amova y el Gst = 0.040 del análisis de correspondencia múltiple. Los valores de similitud genética, el análisis de grupo, el análisis de correspondencia múltiple y el nivel de introgresion, indicaron diferencias significativas (P<0.0001 en los niveles de introgresión. El bajo nivel de diferenciación genética entre poblaciones podría ser el resultado de peces con el mismo origen genético y la alta variación dentro de poblaciones se puede presentar por prácticas de manejo. La introgresión entre piscícolas es significativa para O. aureus, mientras que las especies O. niloticus y O. mosambicus se encuentran introgresadas de forma similar en las poblaciones de Oreochromis sp.

  17. Caracterización de la diversidad genética en naranja y comparación del polimorfismo de microsatélites amplificados al azar (RAMs usando electroforesis de poliacrilamida y agarosa

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    Muñoz Flores Jaime Eduardo

    2009-12-01

    Full Text Available Se compararon las eficiencias de tres métodos de electroforesis en agarosa y poliacrilamida, usando la cámara pequeña de DNA Sequencing System y cámara grande OWL Sequi-Gen Sequencing Cell, en la detección del polimorfismo en 21 accesiones de naranja (Citrus sinensis con empleo del cebador CGA. El gel de poliacrilamida dio mejor resolución de los productos amplificados vía PCR producidos por RAMs. Este permitió una mejor detección de bandas de ADN polimórficas, lo que facilitó la identificación de la variabilidad genética. La electroforesis en agarosa puede ser más conveniente en otras aplicaciones, debido al bajo costo y fácil aplicación. El estudio de diversidad genética en naranja usando microsatélites RAMs diferenció 51 accesiones en siete grupos con 0.75 de similaridad y 0.25 de heterocigosidad, lo que revela bajo polimorfismo genético. La técnica RAMs permitió agrupar las accesiones en Comunes o Blancas, Navel y Pigmentadas o Sanguinas.

  18. Earthquakes, influenza and cycles of Indian kala-azar.

    Science.gov (United States)

    Dye, C; Wolpert, D M

    1988-01-01

    It is suggested that previous data indicate 3 major epidemics of kala-azar in Assam between 1875 and 1950, with inter-epidemic periods of 30-45 and 20 years. This deviates from the popular view of regular cycles with a 10-20 year period. A deterministic mathematical model of kala-azar is used to find the simplest explanation for the timing of the 3 epidemics, paying particular attention to the role of extrinsic (drugs, natural disasters, other infectious diseases) versus intrinsic (host and vector dynamics, birth and death rates, immunity) processes in provoking the second. We conclude that, whilst widespread influenza in 1918-1919 may have magnified the second epidemic, intrinsic population processes provide the simplest explanation for its timing and synchrony throughout Assam. The model also shows that the second inter-epidemic period is expected to be shorter than the first, even in the absence of extrinsic agents, and highlights the importance of a small fraction of patients becoming chronically infectious (with post kala-azar dermal leishmaniasis) after treatment during an epidemic.

  19. Caracterización de tilapia roja (Oreochromis sp. con marcadores moleculares RAPD

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    Muñoz Jaime Eduardo

    2010-06-01

    Full Text Available Se utilizó la técnica RAPD (amplificación al azar de ADN polimórfico para el estudio de la diversidad genética de Oreochromis sp. (tilapia roja en cinco piscícolas del Valle del Cauca (Colombia y en la determinación del nivel de introgresión de las especies parentales Oreochromis mosambicus, O. niloticus y O. aureus. Se evaluaron 25 cebadores, ocho fueron polimórficos y se obtuvieron 109 bandas. Los valores de heterocigosidad esperada (0.196 a 0.256 y la estructura genética (Gst = 0.22 para Oreochromis sp. indicaron un elevado grado de polimorfismo y alta estructuración genética. Estos resultados fueron consistente con el Fst = 0.268 (P < 0.0001 dado por el Amova y el Gst = 0.040 del análisis de correspondencia múltiple. Los valores de similitud genética, el análisis de grupo, el análisis de correspondencia múltiple y el nivel de introgresion, indicaron diferencias significativas (P<0.0001 en los niveles de introgresión. El bajo nivel de diferenciación genética entre poblaciones podría ser el resultado de peces con el mismo origen genético y la alta variación dentro de poblaciones se puede presentar por prácticas de manejo. La introgresión entre piscícolas es significativa para O. aureus, mientras que las especies O. niloticus y O. mosambicus se encuentran introgresadas de forma similar en las poblaciones de Oreochromis sp.

  20. RAPD-based screening of genomic libraries for positional cloning.

    Science.gov (United States)

    Dioh, W; Tharreau, D; Lebrun, M H

    1997-12-15

    RAPD markers are frequently used for positional cloning. However, RAPD markers often contain repeated sequences which prevent genomic library screening by hybridisation. We have developed a simple RAPD analysis of genomic libraries based on the identification of cosmid pools and clones amplifying the RAPD marker of interest. Our method does not require the cloning or characterisation of the RAPD marker as it relies on the analysis of cosmid pools or clones using a simple RAPD protocol. We applied this strategy using four RAPD markers composed of single copy or repeated sequences linked to avirulence genes of the rice blast fungus Magnaporthe grisea . Cosmids containing these RAPD markers were easily and rapidly identified allowing the construction of physical contigs at these loci.

  1. Immunopathology of post kala-azar dermal leishmaniasis (PKDL): T-cell phenotypes and cytokine profile

    DEFF Research Database (Denmark)

    Ismail, A; El Hassan, A M; Kemp, K

    1999-01-01

    In Sudan, post kala-azar dermal leishmaniasis (PKDL) caused by Leishmania donovani develops in half of the patients treated for visceral leishmaniasis (kala-azar). In most patients lesions heal spontaneously, but in others symptoms are severe and persist for years. This study examined...

  2. Post-kala-azar dermal leishmaniasis associated with AIDS

    Directory of Open Access Journals (Sweden)

    Bittencourt Achiléa

    2003-01-01

    Full Text Available Post-kala-azar dermal leishmaniasis (PKDL is rarely reported in South America. In spite of the fact that there are many reports about the association of visceral leishmaniasis and AIDS, PKDL is very uncommon in HIV-positive patients, and so far only four cases have been documented in the literature. We present another case with unusual clinicopathological aspects. The patient, a 28-year-old male, from Salvador, Bahia (an endemic area presented with clinical manifestations of visceral leishmaniasis three years after the diagnosis of AIDS. During treatment for visceral leishmaniasis he developed disseminated miliary papules. Microscopically, the skin biopsy showed a "saw-tooth" pattern with a lichenoid mononuclear infiltrate simulating lichen planus. The histopathological diagnosis was achieved through the finding of amastigotes. The authors discuss the clinicopathological aspects of this case based on a review of the specific literature.

  3. Post-kala-azar dermal leishmaniasis associated with AIDS

    Directory of Open Access Journals (Sweden)

    Bittencourt Achiléa

    2002-01-01

    Full Text Available Post-kala-azar dermal leishmaniasis (PKDL is rarely reported in South America. In spite of the fact that there are many reports about the association of visceral leishmaniasis and AIDS, PKDL is very uncommon in HIV-positive patients, and so far only four cases have been documented in the literature. We present another case with unusual clinicopathological aspects. The patient, a 28-year-old male, from Salvador, Bahia (an endemic area presented with clinical manifestations of visceral leishmaniasis three years after the diagnosis of AIDS. During treatment for visceral leishmaniasis he developed disseminated miliary papules. Microscopically, the skin biopsy showed a "saw-tooth" pattern with a lichenoid mononuclear infiltrate simulating lichen planus. The histopathological diagnosis was achieved through the finding of amastigotes. The authors discuss the clinicopathological aspects of this case based on a review of the specific literature.

  4. Ambisome plus miltefosine for Indian patients with kala-azar.

    Science.gov (United States)

    Sundar, Shyam; Sinha, Prabhat Kumar; Verma, Deepak K; Kumar, Nawin; Alam, Shahnawaj; Pandey, Krishna; Kumari, Poonam; Ravidas, Vidyanand; Chakravarty, Jaya; Verma, Neena; Berman, Jonathan; Ghalib, Hashim; Arana, Byron

    2011-02-01

    The combination of one intravenous administration of 5mg/kg Ambisome and oral administration of miltefosine, 2.5mg/kg/day for 14 days, was evaluated in 135 Indian patients with kala-azar. The Intent-to-Treat cure rate at 6 months was 124 of the 135 enrolled patients (91.9%: 95% CI = 86-96%), and the per protocol cure rate was 124 of 127 evaluable patients (97.6%: 95% CI = 93-100%). Side effects could be attributed to each drug separately: fevers, rigors and back pain due to Ambisome; gastrointestinal side effects due to miltefosine. This combination is attractive for reasons of efficacy, tolerance, and feasibility of administration, although the gastrointestinal side effects of miltefosine require medical vigilance. Clinical Trials.gov identification number: NCT00371995.

  5. Misdiagnosis and Mistreatment of Post-Kala-Azar Dermal Leishmaniasis

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    Ahmed Mohamed El Hassan

    2013-01-01

    Full Text Available Post-kala-azar dermal leishmaniasis (PKDL is a known complication of visceral leishmaniasis (VL caused by L. donovani. It is rare in VL caused by L. infantum and L. chagasi. In Sudan, it occurs with a frequency of 58% among successfully treated VL patients. In the majority of cases, PKDL can be diagnosed on the basis of clinical appearance, distribution of the lesions, and past history of treated VL. The ideal diagnostic method is to demonstrate the parasite in smears, by culture or PCR. Diagnosis is particularly difficult in patients who develop PKDL in the absence of previous history of visceral leishmaniasis. We describe a case of cutaneous leishmaniasis misdiagnosed as PKDL and 3 cases of PKDL who were either misdiagnosed or mistreated as other dermatoses. This caused exacerbation of their disease leading to high parasite loads in the lesions and dissemination to internal organs in one of the patients, who was also diabetic. The latter patient had L. major infection. A fourth patient with papulonodular lesions on the face and arms of 17-year duration and who was misdiagnosed as having PKDL is also described. He turned out to have cutaneous leishmaniasis due to L. major. Fortunately, he was not treated with steroids. He was cured with intravenous sodium stibogluconate.

  6. Performance of Kala-Azar Surveillance in Gaffargaon Subdistrict of Mymensingh, Bangladesh

    Science.gov (United States)

    Rahman, Kazi Mizanur; Samarawickrema, Indira V. M.; Harley, David; Olsen, Anna; Butler, Colin D.; Sumon, Shariful Amin; Biswas, Subrata Kumar; Luby, Stephen P.; Sleigh, Adrian C.

    2015-01-01

    Introduction Elimination of kala-azar is planned for South Asia requiring good surveillance along with other strategies. We assessed surveillance in Gaffargaon upazila (a subdistrict of 13 unions) of Mymensingh district, Bangladesh highly endemic for kala-azar. Methods In 4703 randomly sampled households, within nine randomly sampled villages, drawn from three randomly sampled unions, we actively searched for kala-azar cases that had occurred between January 2010 and December 2011. We then searched for medical records of these cases in the patient registers of Gaffargaon upazila health complex (UHC). We investigated factors associated with the medical recording by interviewing the cases and their families. We also did a general observation of UHC recording systems and interviewed health staff responsible for the monthly reports of kala-azar cases. Results Our active case finding detected 58 cases, but 29 were not recorded in the Gaffargaon UHC. Thus, only 50% (95% CI: 37%–63%) of kala-azar cases were reported via the government passive surveillance system. Interviews with health staff based in the study UHC revealed the heavy reporting burden for multiple diseases, variation in staff experience, high demands on the staff time and considerable complexity in the recording system. After adjusting for kala-azar treatment drug, recording was found more likely for those aged 18 years or more, males, receiving supply and administration of drug at the UHC, and more recent treatment. Discussion Fifty percent of kala-azar cases occurring in one highly endemic area of Bangladesh were recorded in registers that were the source for monthly reports to the national surveillance system. Recording was influenced by patient, treatment, staff and system factors. Our findings have policy implications for the national surveillance system. Future studies involving larger samples and including interviews with health authorities at more central level and surveillance experts at the

  7. Performance of kala-azar surveillance in Gaffargaon subdistrict of Mymensingh, Bangladesh.

    Directory of Open Access Journals (Sweden)

    Kazi Mizanur Rahman

    2015-04-01

    Full Text Available Elimination of kala-azar is planned for South Asia requiring good surveillance along with other strategies. We assessed surveillance in Gaffargaon upazila (a subdistrict of 13 unions of Mymensingh district, Bangladesh highly endemic for kala-azar.In 4703 randomly sampled households, within nine randomly sampled villages, drawn from three randomly sampled unions, we actively searched for kala-azar cases that had occurred between January 2010 and December 2011. We then searched for medical records of these cases in the patient registers of Gaffargaon upazila health complex (UHC. We investigated factors associated with the medical recording by interviewing the cases and their families. We also did a general observation of UHC recording systems and interviewed health staff responsible for the monthly reports of kala-azar cases.Our active case finding detected 58 cases, but 29 were not recorded in the Gaffargaon UHC. Thus, only 50% (95% CI: 37%-63% of kala-azar cases were reported via the government passive surveillance system. Interviews with health staff based in the study UHC revealed the heavy reporting burden for multiple diseases, variation in staff experience, high demands on the staff time and considerable complexity in the recording system. After adjusting for kala-azar treatment drug, recording was found more likely for those aged 18 years or more, males, receiving supply and administration of drug at the UHC, and more recent treatment.Fifty percent of kala-azar cases occurring in one highly endemic area of Bangladesh were recorded in registers that were the source for monthly reports to the national surveillance system. Recording was influenced by patient, treatment, staff and system factors. Our findings have policy implications for the national surveillance system. Future studies involving larger samples and including interviews with health authorities at more central level and surveillance experts at the national level will generate

  8. Current treatment of visceral leishmaniasis (Kala-azar: an overview

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    Premshanker S. Singh

    2014-06-01

    Full Text Available Visceral Leishmaniasis (VL is also popularly known as kala-azar which was first reported in early forties and since then it continues to affect millions of people. The ranges of common drugs available for the treatment of visceral leishmaniasis are limited. It mainly includes pentavalent antimonials e.g. stibogluconate (SbV, amphotericin B deoxycholate (AB, lipid formulations of amphotericin B (L-AB, miltefosine (MF and paromomycin (PM - all of which have limitations in terms of toxicity, variable efficacy, price and inconvenient treatment schedules. Most are parenteral except MF which is administered orally. Due to the parasite and #8217;s drug resistance, the most widely used (SbV of these drugs is now of little use in northern Bihar, India, which alone accounts for 50% of the worlds burden of visceral leishmaniasis. In areas of resistance to SbV, AB is highly effective. The formulation of AB in liposomes (L-AB has been a major advancement in the treatment of visceral leishmaniasis. However, despite a significant reduction in price, this treatment remains very expensive for endemic countries like India. Combination short course therapy has been reported by many researchers who found that it is equally effective as conventional monotherapy with added benefits of less side effects, better compliance and less resistance. The aim of this article is to review the current aspects of the treatment for leishmaniasis, giving an overview of current agents clinically used to new agents and modalities of treatment under development. [Int J Res Med Sci 2014; 2(3.000: 810-817

  9. Authentication of shankhpushpi by RAPD markers

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    Showkat Hussain Ganie

    2012-05-01

    Full Text Available Background: “Shankhpushpi”, an important indigenous drug of Ayurveda, an ancient system of Indian medicine, improves memory power and intellect. It is used in many Ayurvedic formulations, either singly or in combination with other herbs, meant for sleeplessness, epilepsy, hallucinations and anxiety. At least three different plant species viz., Clitoria ternatea, Convolvulus pluricaulis and Evolvulus alsinoides are as the source of this drug in the different parts of the country. Because of increased demand and high price, shankhpushpi is often adulterated in the trade by other related species. Therefore, a reliable authentication method is needed to facilitate differentiation/identification of the genuine material from its adulterants. The present study was aimed at developing RAPD-based markers for identification of C. pluricaulis, E. alsinoides and C. ternatea, and analyzing the market samples of the drug to ascertain their authenticity. Material and Methods: Fresh samples of source plants of shankhpushpi were collected from Ghaziabad and Delhi. The market samples were procured from the crude-drug markets of different geographical regions of India. The amplified polymorphic DNA (RAPD technique was employed for characterization of genuine and market samples. Twenty-five 11-mer oligonucleotide primers were used to amplify the DNA isolated. Results: Out of 25 primers, only four (OPN-03, OPN-04, OPN-05 and OPN-06 yielded amplification products that produced clear and reproducible bands, which were used to characterize the market samples. RAPD profile of some market samples did not match with the authentic samples, indicating that these samples were either adulterated or spurious. Conclusion: The RAPD markers developed in this study may provide guidance for the authentication of plant materials traded as shankhpushpi.

  10. The molecular marker of antagonistic genes of biological bacteria against rice sheath blight by RAPD

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    @@Forrty-one isolates of Bacillus amyloliquefaciens were differentiated from 184 G + bacterial strains having genetic similarities over 75%based on BOX-PCR fingerprint. Antagonism against to Rhizotonia solani in vitro was tested.Four isolates of B. arayloliquefaciens (2 isolates with antagonistic ability, G 396 + and G229 +, and 2 isolates without antagonistic ability, G433-and G434-) were selected to screen effective primers for RAPD analysis. Of 124 random primers (AA, AB, AC, AD, AE, AM, and AL) tested.

  11. DIFFERENT RESULTS BY DIFFERENT COMMERCIAL TAQ DNA POLYMERASE IN RAPD

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    @@ RAPD (Random Amplified Polymorphic DNA) technique has been widely used in animal, plant, human and microorganism research since it was first established by Williams in 1990[1-3]. But, because of low annealed temperature and short 10-nt primers, the resolution and repetition is low in RAPD. The stability of RAPD is influenced by many factors such as the concentration of template, primers, dNTP, Mg++,and Taq DNA polymerase[4-6]. The influence on amplified products of different commercial Taq DNA polymerase in RAPD was studied in this paper.

  12. Genetic diversity ofUstilago hordei in Tibetan areas as revealed by RAPD and SSR

    Institute of Scientific and Technical Information of China (English)

    ZHOU Yu; CHAO Gui-mei; LIU Jia-jia; ZHU Ming-qi; WANG Yang; FENG Bai-li

    2016-01-01

    Covered smut, which is caused byUstilago hordei(Pers.) Lagerh., is one of the most damaging diseases of highland barley (Hordeum vulgare Linn. var. nudum Hook. f) in Tibetan areas of China. To understand the molecular diversity ofU. hordei, a total of 27 isolates, which were colected from highland barley plants from Tibet, Sichuan, Qinghai, and Gansu provinces/autonomous region, were analyzed using random ampliifed polymorphic DNA (RAPD) and simple sequence repeat (SSR) markers. Among the 100 RAPD primers used, 24 primers exhibited polymorphism. A total of 111 fragments were ampliifed, of which 103 were polymorphic with a polymorphic rate of 92.79%. The average observed number of aleles (Na), effective number of aleles (Ne), Nei’s genetic diversity (H), Shannon’s information index (I) and polymorphism information content (PIC) value in the RAPD markers were 1.9279, 1.5016, 0.2974, 0.4503 and 0.6428, respectively. For the SSR markers, 40 of the 111 primer pairs exhibited polymorphism and provided a total of 119 bands, of which 109 were polymorphic and accounted for 91.60% of the total bands. TheNa,Ne,H,I andPIC values of the SSR markers were 1.9160, 1.4639, 0.2757, 0.4211 and 0.4340, respectively. The similarity coefifcients ranged from 0.4957 to 0.9261 with an average of 0.7028 among al the 27 isolates used. The dendrogram, which was developed based on the RAPD and SSR combined marker dataset showed that the 27U. hordei isolates were divided into 3 clusters at similarity coefifcient of 0.7314. We determined that RAPD and SSR markers can be successfuly used to assess the genetic variation amongU. hordei isolates. The RAPD markers revealed higher levels of genetic polymorphism than did the SSR markers in this study. There existed a moderate genetic difference among isolates. The molecular variation and differentiation was somewhat associated with geographical origin but not for al of the isolates.

  13. Genetic Diversity in Upland Cotton(Gossypium hirsutum L. )Cultivars Based on RAPDs and SSRs%基于RAPD和SSR标记分析的陆地棉品种遗传多样性

    Institute of Scientific and Technical Information of China (English)

    Long-fu ZHU; Xian-long ZHANG; Yi-chun NIE

    2002-01-01

    @@ Quantification and classification of diversity in germplasm collection is important for both genetic researchers and plant breeders. Some advance was made in this area in the world ( Liu et al, 2000) based on SSRs and in China(Xu et al, 2001; 2002 ) based on RAPDs. In this research, 72 cultivars including 14 latest introduced and 30 Bt-transformed ones were evaluated by RAPDs and SSRs.

  14. Subspecies specific RAPD markers in rice

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    The cultivated rice (Oryza sativa L.) is known to contain two major subspecies, indica (O. sativa L. ssp. indica) and japonica (O. sativa L. ssp. japonica). The indica and japonica differentiation resulted in significance of hybrid sterility and hybrid breakdown, which are barriers of gene flow between the two major subgene pools within O. sativa. Traditional classification of indica and japonica germplasm based on isozymes. Here, we report the identification of several random amplified polymorphic DNA (RAPD) markers that have alleles specific in indica or japonica varieties and thus provide a quick and accurate tool to distinguish japonica lines from indica ones.

  15. La demanda de lotería y juegos de azar: Su realidad y sus potencialidades

    Directory of Open Access Journals (Sweden)

    Jorge Iván González

    2006-12-01

    Full Text Available El estudio sobre [as loterías y los juegos de azar tiene implicaciones de muy diversa naturaleza. Toca aspectos básicos de la teoría de la elección bajo riesgo y, ademas, tiene implicaciones fiscales muy importantes. La forma de administrar el monopolio de las loterías y juegos de azar plantea retos muy interesantes en el campo de la gerencia y de la hacienda pública Después de presentar el monto de las ventas de las principales loterías, analizamos el significado de la demanda consuetudinaria. En la sección siguiente evaluamos los determinantes del precio óptimo Posteriormente describimos el perfil del comprador de loterias y juegos de azar, para finalizar el ensayo con unas reflexiones sobre el impacto fiscal de las loterías.

  16. Post-kala-azar dermal Leishmaniasis in two different clinical contexts*

    Science.gov (United States)

    Barroso, Daniel Holanda; Silva, Claúdia Elise Ferraz; Vasconcelos, Ana Carolina Depes Perdigao e; Cavalcanti, Silvana Maria de Morais; de Brito, Maria Edileuza Felinto; Medeiros, Angela Cristina Rapela

    2015-01-01

    In Brazil, visceral Leishmaniasis is caused by Leishmania chagasi. The development of cutaneous lesions in visceral leishmaniasis patients has been described in two different clinical contexts. Patients with compromised immunity can develop skin lesions as a direct consequence of a current visceral disease. Equally, patients with a history of kala-azar and progressive, immune improvement occasionally develop skin lesions as a consequence of immune reconstitution infl ammatory syndrome. These cases manifest in similar fashion to the classic form of post-kala-azar dermal Leishmaniasis. We describe different cases that exemplify these two clinical presentations. PMID:26312689

  17. Detección “in vivo” mediante RAPD de alteraciones en el ADN producidas por benzo(apireno

    Directory of Open Access Journals (Sweden)

    C Becerril

    2004-01-01

    Full Text Available La técnica de RAPD (Amplificación al Azar de ADN Polimórfico permite detectar alteraciones inespecíficas en el ADN procedente de células que poseen una dotación genética idéntica, como son las líneas celulares establecidas, mediante la comparación del patrón de bandas de las células expuestas y no expuestas a la acción de genotóxicos. En los últimos años hemos desarrollado una metodología sensible y reproducible utilizando la línea celular RTG-2, derivada de trucha arco iris (Oncorhynchus mykiss. Sin embargo, es preciso comprobar la capacidad predictiva de este ensayo mediante estudios in vivo. La línea celular RTG-2, como se ha evidenciado en trabajos anteriores, presenta una gran similitud genética con la especie de la que procede. Por ello, en este trabajo, se ha llevado a cabo una exposición subletal a benzo(apireno mediante inyección intraperitoneal de 69 μg/g de p.c. en alevines de trucha arco iris, valorando la aparición de mutaciones mediante la comparación del patrón de bandas obtenido a partir del ADN de células de sangre periférica, a diferentes tiempos (1 - 3 meses. Debido a que la presencia de bandas polimórficas dificulta el análisis entre los grupos de individuos tratados y no tratados, las comparaciones se realizaron en un mismo individuo antes y después del tratamiento. Los análisis cualitativos y cuantitativos mostraron tanto la aparición de nuevas bandas, como alteraciones en su intensidad confirmando, de esta manera, los resultados que previamente habíamos obtenido in vitro tras exposiciones a este mismo genotóxico

  18. Fronteras del corazón shamánico: Azares y dilemas Qom

    Directory of Open Access Journals (Sweden)

    Pablo Wright

    2010-07-01

    Full Text Available En este artículo analizo dilemas y paradojas que plantea la modernidad en el caso de los Takshek Qom o tobas del oriente de Formosa (Argentina. Para ello me centro en la experiencia del lkillakte o corazón como uno de los núcleos vitales y existenciales de la persona. Allí se alojan las distintas clases de poder shamánico y también es sede de los pensamientos y las emociones. Me interesa explorar qué le pasa en términos simbólicos al corazón shamánico frente al contacto con el catolicismo y el evangelismo, y sus definiciones de lo sagrado y el poder numinoso, la enfermedad y la terapia. De este modo es posible observar al corazón como un lugar-proceso que ha transformado sus fronteras, redefiniendo su anatomía y fisiología ontológicas. Así, como un operador metonímico sensible, los azares del corazón shamánico condensan eventos, símbolos y metáforas significativas de la historia qom.In this paper I analyze certain dilemmas and paradoxes of modernity in the case of the Takshek Qom or Eastern Toba (Formosa province, Argentina. To do so I focus on the experience of the heart or lkillakte, defined as the person's key vital and existential center. Within it thoughts and emotions occur, and it also houses different kinds of shamanic power. From a symbolic approach, I explore what happens with the shamanic heart under the influence of Catholicism and Evangelism, stressing their definitions of the sacred and numinous power, illness and therapy. In this sense, I suggest to regard the heart as a place-process that has widened its boundaries, redefining its anatomy and ontological physiology. Thus, as a sensible, metonymic operator, I argue that the contingencias of shamanic hearts condense events, symbols and meaningful metaphors of Qom history.

  19. CHARACTERIZATION OF A NEW BIOTYPE Moringa OF SAUDI ARABIA USING RAPD AND ISSR MARKERS

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    Iyan Robiansyah

    2015-11-01

    Full Text Available Moringa peregrina and M. oleifera are the only Moringa species found in Saudi Arabia. Both species are drought resistant and have very high nutritional and medicinal properties. Detection of genetic diversity is of great value for the improvement of nutritional and medicinal value of these plants. The aim of the present study was to characterize a new biotype Moringa observed in Al Bahah Region, Saudi Arabia. We used 11 RAPD and 15 ISSR primers to characterize and compare the new biotype with M. peregrina and M. oleifera. Level of polymorphism generated by each marker was calculated. We also calculate Nei and Li’s coefficient to measure the genetic distance between the studied species. Level of polymorphism generated by RAPD and ISSR was 46% and 57%, respectively. RAPD and ISSR primers revealed that the new biotype shared 55 amplicons (45.08% with both M. peregrina and M. oleifera, 28 amplicons with M. peregrina (22.95%, 21 amplicons (17.21% with M. oleifera, and displayed 18 unshared amplicons (14.75%. Based on RAPD data, genetic distance between M. oleifera and M. peregrina was 0.32, whereas genetic distance between the new biotype and M. oleifera and M. peregrina was 0.21 and 0.29, respectively. For ISSR data, genetic distance between M. oleifera and M. peregrina was 0.5, whereas genetic distance between the new biotype and M. oleifera and M. peregrina was 0.36 and 0.34, respectively. Based on these results we suggested that the new biotype is a hybrid crossbred between M. peregrina and M. oleifera.

  20. Linnamaastike kaitse on osa arengust / Jukka Jokilehto, Mehr Azar Soheil ; intervjueerinud Liisa Pakosta

    Index Scriptorium Estoniae

    Jokilehto, Jukka

    2010-01-01

    Vestlus soome arhitekti, Nova Gorica ülikooli professori, UNESCO ja ICOMOSi konsultandi Jukka Jokilehto ja tema abikaasa, pärslannast arhitekti Mehr Azar Soheiliga. Ka Tallinna vanalinnast maailmapärandi komitees valmivate linnamaastike kaitseks mõeldud soovituste valguses, linna arengust sadama suunas. Eestikeelsena ilmus J. Jokilehto raamat "Arhitektuuri konserveerimise ajalugu" (Tallinn : Eesti Kunstiakadeemia, 2010)

  1. The reproducibility of RAPD profiles: Effects of PCR components on RAPD analysis of four centaurium species

    Directory of Open Access Journals (Sweden)

    Skorić Marijana

    2012-01-01

    Full Text Available Random amplified polymorphic DNA (RAPD analysis is a simple and reliable method used to detect DNA polymorphism. Several factors can affect the amplification profiles, thereby causing false bands and non-reproducibility of the assay. In this study, we analyzed the effects of different concentrations of primer, magnesium chloride, template DNA and Taq DNA polymerase to develop and standardize a RAPD protocol for Centaurium species. The optimized PCR reaction mixture included: 50 ng of DNA extracted using a CTbased protocol, 2.5 mM MgCl2, 7.5 pmol primer and 2 U of Taq polymerase in a final volume of 25 μl. Each of the five primers used in experiments (OPB11, OPB15, OPB18, OPF05 and OPH02 generated reproducible and distinguishable fingerprinting patterns of four Centaurium species. The obtained optimized RAPD protocol and the selected primers are useful for our further work in the genetic diversity studies of Centaurium species.

  2. Caracterización de la diversidad genética en naranja y comparación del polimorfismo de microsatélites amplificados al azar (RAMs usando electroforesis de poliacrilamida y azarosa Characterization of the genetic diversity in orange, and comparison of polymorphism in randomly-amplifed microsatellites (RAMs, using polyacrylamide and agarose electrophoresis

    Directory of Open Access Journals (Sweden)

    Ana Cruz Morillo Coronado

    2009-10-01

    Full Text Available Se compararon las eficiencias de tres métodos de electroforesis en agarosa y poliacrilamida, usando la cámara pequeña de DNA Sequencing System y cámara grande OWL Sequi-Gen Sequencing Cell, en la detección del polimorfismo en 21 accesiones de naranja (Citrus sinensis con empleo del cebador CGA. El gel de poliacrilamida dio mejor resolución de los productos amplificados vía PCR producidos por RAMs. Este permitió una mejor detección de bandas de ADN polimórficas, lo que facilitó la identificación de la variabilidad genética. La electroforesis en agarosa puede ser más conveniente en otras aplicaciones, debido al bajo costo y fácil aplicación. El estudio de diversidad genética en naranja usando microsatélites RAMs diferenció 51 accesiones en siete grupos con 0.75 de similaridad y 0.25 de heterocigosidad, lo que revela bajo polimorfismo genético. La técnica RAMs permitió agrupar las accesiones en Comunes o Blancas, Navel y Pigmentadas o Sanguinas.We compared the efficiency of three methods of agarose and polyacrylamide electrophoresis (using the small tank of the DNA Sequencing System and the large OWL Sequi-Gen Sequencing Cell, for the detection of polymorphism in 21 accessions of orange (Citrus sinensis, using the primer CGA. The polyacrylamide gel gave better resolution of the PCR-amplified RAM products. This method allowed better detection of polymorphic DNA bands, facilitating the identification of genetic variability. The agarose electrophoresis may be more convenient in other applications, due to its low cost and easy implementation. The study of genetic diversity in orange using RAMs separated 51 accessions into seven groups with 0.75 similarity, and 0.25 heterozygosity, revealing low genetic polymorphism. The RAMs technique grouped the accessions into “Common or White”, “Navel” and “Pigmented or “Sanguine”.

  3. Caracterización de bacterias aisladas en larvas de p.v. mediante rapds

    OpenAIRE

    Cedeño, Ricardo

    2003-01-01

    Caracterización de bacterias aisladas en larvas de P.V. mediante RAPDs La “Vibriosis luminiscente” reportada por varios laboratorios de larvas del Ecuador es asociada principalmente a la bacteria Vibrio harveyi. Los métodos de identificación bacteriana tradicionales se basan en la observación de características morfológicas y pruebas bioquímicas, sin embargo la identificación de especies bacterianas y en especial de Vibrios ambientales ha presentado ciertas dificultades debido al tiempo qu...

  4. Role of remote sensing, geographical information system (GIS) and bioinformatics in kala-azar epidemiology.

    Science.gov (United States)

    Bhunia, Gouri Sankar; Dikhit, Manas Ranjan; Kesari, Shreekant; Sahoo, Ganesh Chandra; Das, Pradeep

    2011-11-01

    Visceral leishmaniasis or kala-azar is a potent parasitic infection causing death of thousands of people each year. Medicinal compounds currently available for the treatment of kala-azar have serious side effects and decreased efficacy owing to the emergence of resistant strains. The type of immune reaction is also to be considered in patients infected with Leishmania donovani (L. donovani). For complete eradication of this disease, a high level modern research is currently being applied both at the molecular level as well as at the field level. The computational approaches like remote sensing, geographical information system (GIS) and bioinformatics are the key resources for the detection and distribution of vectors, patterns, ecological and environmental factors and genomic and proteomic analysis. Novel approaches like GIS and bioinformatics have been more appropriately utilized in determining the cause of visearal leishmaniasis and in designing strategies for preventing the disease from spreading from one region to another.

  5. AN ASSESSMENT ON THE PREVALENCE OF KALA - AZAR IN MUNICIPAL AREA OF KATIHAR DISTRICT, BIHAR

    Directory of Open Access Journals (Sweden)

    Prawin

    2015-04-01

    Full Text Available INTRODUCTION: Kala - azar is a chronic infectious disease of the reticulo - endothelial system , characterized by irregular fever of long duration , anemia , leucopenia , hepato - spleenomegaly and progressive emaciation . It has major medical , psychological , and financial implications and remains a serious public health problem in Bihar . OBJECTIVES: To find the prevalence of kala - azar and its associated factors & also to formulate preventive measures for the control of this disease . MATERIALS AND METHODS: This was a community based cross - sectional study , conducted in 5 municipal wards of Katihar district , from 1 st Jan . 2014 to 31 st Dec . 2014 . Study population was selected by stratified random sampling method . From each ward families were selected by simple random s ampling . After the collection of data using pre - designed and pre - tested Performa , prevalence of kala - azar in municipal area of Katihar district was studied . RESULTS: Among the total study population of 2095 people , 1089 were males and 1006 females . Maximum number of cases ( 60% in our study was between 20 - 29 years of age group and 80% of them were males . Sixty percent ( 60% of the cases belonged to low socio - economic status and 80% cases occurred in persons living in kutccha houses , where the walls of the r oom were made of mud . In 80% of the cases , distance of the cattle shed from houses was less than 15 meters . CONCLUSION: Young adults are more susceptible to kala - azar and the incidence gradually decline with age . Male preponderance may be due to some pract ices of adult males like sleeping in the cattle sheds or close to cattle sheds . The high incidence of the disease among low socio - economic group may be due to insanitary habits and surroundings which provide excellent breeding grounds and shelter for the sand fly .

  6. Avaliação por RAPD de plantas de abacaxizeiro cultivar Smooth Cayenne derivadas do seccionamento do talo e cultura de tecidos Evaluation of pineapple plants cultivar Smooth Cayenne from peduncle division and tissue culture by RAPD

    Directory of Open Access Journals (Sweden)

    MARIA VITÓRIA CECCHETTI GOTTARDI

    2002-04-01

    Full Text Available Foram coletadas, em área comercial da fazenda Córrego dos Bois, município de Canápolis -- MG, plantas de abacaxizeiro cultivar Smooth Cayenne, para serem avaliadas quanto à propagação pelo método do seccionamento do talo e cultura de tecidos, bem como análise por RAPD das mudas decorrentes destes dois processos de propagação. A propagação pelo seccionamento do talo foi eficiente na produção de mudas, tanto em quantidade como em qualidade, em um curto espaço de tempo, além de apresentar a mesma característica genotípica (análise por RAPD das plantas-matrizes de origem. Já no processo de produção de mudas por cultura de tecidos, não foi obtida uma quantidade suficiente de mudas que comprovasse a utilização de uma metodologia mais sofisticada. Além da perda por contaminação em laboratório de 70% do material em estudo, foi necessária a utilização de um longo período, aproximadamente 18 meses, para a obtenção das mudas. Na análise por RAPD das plantas decorrentes deste processo de propagação, foram observados padrões de bandas diferentes em algumas amostras, as quais podem estar relacionadas com uma possível variação somaclonal.Plants of pineapple Smooth Cayenne cultivar were collected from a commercial area, Córrego dos Bois farm, in Canápolis, state of Minas Gerais, to evaluate two different processes of propagation, peduncle division and tissue culture. These same plants were characterized by RAPD analysis. According to the two propagation method, the peduncle division was efficient in seedlings propagation, in quantity as well as in quality, at a short period of time, resulting the same phenotype and genetic characteristics (RAPD analysis from the matrix descent plants. In the tissue culture method, the quantity of seedlings obtained were not good enough that could prove the use of a sophisticated methodology. Besides loosing 70% of the material, it was necessary to expend a long period of time, at

  7. Identification of Tsuga Germplasm by Morphological Characters and RAPD Markers

    Institute of Scientific and Technical Information of China (English)

    ROH Mark S; DE BENEDETTI Laura; JOUNG Young Hee; LEE Nam Sook

    2007-01-01

    Germplasm collection is important to preserve and maximize genetic diversity for germplasm conservation. Tsuga dumosa (D. Don) Eichler in Engler & Prantl. and T. chinensis var. forrestii (Downie) Silba germplasm was collected from three localities in China: Mt. Yulong, Wenfeng Temple and Mt. Dishiergu, Yunnan Province. Accessions were identified based on morphological characters and RAPD markers. The shapes of the apices and margins of needles were examined, and the length and width of needles, cones and seeds from accessions of mature plants were used to compare the morphological differences and to identify the germplasm. Molecular markers generated by randomly amplified polymorphic DNA (RAPD) were also used to characterize the taxa. Although the clustering based on RAPD markers was inconsistent with the morphological characters of the needles, based on the overall morphological characters and on RAPD markers, the accessions from Mt. Yulong and Wenfeng Temple were identified as T. chinensis var. forrestii, and those from Mt. Dishiergu identified as T. dumosa. Taxonomic identification of the accessions was made based on morphology and by RAPD markers concurred. The results indicate that the shapes of the apices and margins of needles particularly from young plants could not be used as a possible key to identify T. dumosa and T. chinensis var. forrestii. Fig 6, Tab 3, Ref 24

  8. Leggere i Classici in Oriente. Il mito della letteratura occidentale in Dai Sijie, Murakami Haruki, Azar Nafisi

    Directory of Open Access Journals (Sweden)

    Niccolò Scaffai

    2011-11-01

    Full Text Available Il saggio prende in considerazione tre opere letterarie recenti (appartenenti tanto alla fiction quanto al genere del non-fiction novel, scritte da altrettanti autori di origini asiatiche: il franco-cinese Dai Sijie, l’iraniana Azar Nafisi e il giapponese Murakami Haruki. In ciascun titolo dei loro rispettivi bestseller internazionali (Balzac e la piccola sarta cinese, Leggere Lolita a Teheran, Kafka sulla spiaggia troviamo un ossimoro dello stesso tipo: il  nome di un autore occidentale canonico viene impiegato in un contesto insolito, legato alla geografia del vicino o dell’estremo Oriente. Ne consegue un effetto di straniamento che finisce per ridare forza alla percezione dell’opera letteraria, forse con maggiore decisione di quanto non sia possibile fare oggi in Occidente. Attraverso l’uso (e il rovesciamento del concetto di ‘orientalismo’ di Edward Said e, per Kafka sulla spiaggia, attraverso l’applicazione di principi della teoria di Lacan, lo studio analizza le dinamiche del trasferimento di miti letterari europei nelle culture orientali.

  9. Genetic diversity analysis in Piper species (Piperaceae) using RAPD markers.

    Science.gov (United States)

    Sen, Sandeep; Skaria, Reby; Abdul Muneer, P M

    2010-09-01

    The genetic diversity of eight species of Piper (Piperaceae) viz., P. nigrum, P. longum, P. betle, P. chaba, P. argyrophyllum, P. trichostachyon, P. galeatum, and P. hymenophyllum from Kerala state, India were analyzed by Random amplified polymorphic DNA (RAPD). Out of 22 10-mer RAPD primers screened, 11 were selected for comparative analysis of different species of Piper. High genetic variations were found among different Piper species studied. Among the total of 149 RAPD fragments amplified, 12 bands (8.05%) were found monomorphic in eight species. The remaining 137 fragments were found polymorphic (91.95%). Species-specific bands were found in all eight species studied. The average gene diversity or heterozygosity (H) was 0.33 across all the species, genetic distances ranged from 0.21 to 0.69. The results of this study will facilitate germplasm identification, management, and conservation.

  10. Analysis of 50 Cucumber Accessions by RAPD Markers

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Genetic diversity of 50 cucumber (Cucumis sativus L.) accessions from American, Holland, Japan and China was detected and evaluated using RAPD markers. 25 selected primers produced 215 scorable polymorphic RAPD bands and the ratio of polymorphism is 86.98%. Four main groups of cucumber accessions could be distinguished from UPGMA analysis: Occidental cucumber, South-China cucumber, North-China cucumber and Xishuangbanna cucumber. Our result confirmed that cucumber is a narrow-based germplasm, nevertheless RAPD analysis was still useful in cucumber genotypic differentiation. More significantly, Xishuangbanna cucumber (Cucumis sativus L. Var Xishuangbannanesis Qi et Yuan), a mutation of cucumber, was clustered together to a special group in the study, which implied its special status in cucumber germplasm.

  11. RAPD analysis of genetic relationships among Sphaeropsis sapinea isolates

    Institute of Scientific and Technical Information of China (English)

    WU Xiaoqin; XIONG Dabin; WANG Yu

    2007-01-01

    Genetic relationships were studied among 23 isolates of Sphaeropsis sapinea collected from China,the United States,England,South Africa and Chile by using a random amplification of a polymorphic DNA (RAPD) analytical method.One hundred and 35 DNA fragments were amplified with 12 random primers by a polymerase chain reaction PCR technique and 96.3% were polymorphic.The genetic dendrogram based on RAPD analysis showed that the S.sapinea isolates could be divided into three types.Isolate CWS41 from Chile was separated genetically as the first type that was different from other isolates and isolates F2 and J2 from China comprised the second group.The third RAPD group accommodated other isolates including the B morphotype isolate CWS43 from the United States.

  12. Spatial and temporal variation and hotspot detection of kala-azar disease in Vaishali district (Bihar, India

    Directory of Open Access Journals (Sweden)

    Bhunia Gouri Sankar

    2013-02-01

    Full Text Available Abstract Background An improved understanding in transmission variation of kala-azar is fundamental to conduct surveillance and implementing disease prevention strategies. This study investigated the spatio-temporal patterns and hotspot detection for reporting kala-azar cases in Vaishali district based on spatial statistical analysis. Methods Epidemiological data from the study area during 2007–2011 was used to examine the dynamic space-time pattern of kala-azar outbreaks, and all cases were geocoded at a village level. Spatial smoothing was applied to reduce random noise in the data. Inverse distance weighting (IDW is used to interpolate and predict the pattern of VL cases distribution across the district. Moran’s I Index (Moran’s I statistics was used to evaluate autocorrelation in kala-azar spatial distribution and test how villages were clustered or dispersed in space. Getis-Ord Gi*(d was used to identify the hotspot and cold spot areas within the study site. Results Mapping kala-azar cases or incidences reflects the spatial heterogeneity in the incidence rate of kala-azar affected villages in Vaishali district. Kala-azar incidence rate map showed most of the highest endemic villages were located in southern, eastern and northwestern part of the district; in the middle part of the district generally show the medium occurrence of VL. There was a significant positive spatial autocorrelation of kala-azar incidences for five consecutive years, with Moran’s I statistic ranging from 0.04-0.17 (P P Conclusions The results pointed to the usefulness of spatial statistical approach to improve our understanding the spatio-temporal dynamics and control of kala-azar. The study also showed the north-western and southern part of Vaishali district is most likely endemic cluster region. To employ exact and geographically suitable risk-reduction programmes, apply of such spatial analysis tools should suit a vital constituent in epidemiology research

  13. DNA Extraction from Eriocaulon Plants and Construction of RAPD System

    Institute of Scientific and Technical Information of China (English)

    Xue Xian; Lin Shanzhi; Zhang Zhixiang

    2004-01-01

    There have been many arguments on the classification of Eriocaulon Linn. by morphology so far, and little is known about the use of molecular marker for genetic for genetic diversity of Eriocaulon plants. To apply the technique of molecular marker to the research of genetic diversity of Eriocaulon plants, the study of the extraction method of DNA from the Eriocaulon plants and the RAPD system are essential for researchers. In this paper, the extraction of genome DNA from the silica-gel-dried leaves of several species of Eriocaulon distributed in China was studied, and the best RAPD analysis technique condition of Eriocaulon plants was analyzed.

  14. A clinicopathologic study on kala azar:report of 3 cases diagnosed by bone marrow smear eXamination%骨髓穿刺诊断黑热病3例及临床病理分析

    Institute of Scientific and Technical Information of China (English)

    柳玮华; 周小鸽; 张淑红

    2014-01-01

    目的:报道黑热病累及骨髓3例,探讨黑热病的组织学特点、诊断、鉴别诊断。方法对3例黑热病患者的骨髓穿刺标本行 HE、PAS、免疫组化染色,结合临床表现进行光镜分析,并进行文献复习。结果利什曼原虫的确诊主要依靠病原学检查。结论黑热病目前的患病率相对较低,如果患者来自疫区,有高热反复不退的症状需考虑黑热病的可能性,需要注意与组织胞浆菌和马尔尼菲青霉菌进行鉴别,本病预后较好,大部分可治愈。%Objective To explore the clinical histological features,diagnosis and differential diagnosis of kala azar. Methods The bone marrow smears of patients with kala azar were examined by light microscopy after HE staining,and related literatures had been reviewed. Results The final diagnosis of kala azar depended upon:the finding of pathogenic protozoa,the amastigotes of Leishmania. Conclusion The incidence rate of kala azar in China is low,it should be considered when patients with long term fever coming from endemic area. It′s hard to make differenti-al diagnosis with histoplasmosis and infection with Penicillium marneffei. The prognosis of this disease is good,most of patients can be cured by chemotherapy.

  15. A Study on Health Seeking Behaviors of Patients of Post-Kala-Azar Dermal Leishmaniasis

    Directory of Open Access Journals (Sweden)

    Ariful Basher

    2015-01-01

    Full Text Available Post-Kala-Azar Dermal Leishmaniasis (PKDL remains a major public health threat in Bangladesh. A cross-sectional study was carried out in Surya Kanta Kala azar Research Centre (SKKRC, Mymensingh, from January 2012 to July 2013 to evaluate the health seeking behaviour and the length of delay of PKDL management. The consecutive 200 diagnosed PKDL cases that got treatment in SKKRC hospital were subjected to evaluation. Most (98% of the patients were not aware and had no knowledge about PKDL, though 87.5% had a history of history of Kala-azar treatment. Many patients reported first to village doctor (15.5%, the pharmacy shop (10%, or traditional health provider (7.5% upon recognition of symptom. The time between the initial symptom recognition and first medical consultation (patient delay ranged from 10 days to 4745 days (13 years with a median of 373 days (mean: 696; IQR: 138 to 900 days. The time between first medical consultations to definite treatment (system delay ranged from 0 days to 1971 days (5.4 years, with a median delay of 14 days (mean: 46.48; IQR: 7 to 44 days that was reported in this study. Age, education, occupation, and residential status had significant association with patient delay (P<0.05. Educational status, occupation, number of treatment providers, and first health care provider had a significant association with system delay (P<0.05. Success in PKDL diagnosis and treatment requires specific behavior from patients and health care providers which facilitate those practices.

  16. Entre la ley y el azar: La trama vincular del mundo político-empresarialde la frontera sur pampeana en el siglo XIX

    Directory of Open Access Journals (Sweden)

    Andrea Reguera

    2006-12-01

    Full Text Available Entre la ley y el azar hace referencia a un momento muy particular de nuestra historia argentina, el de la construcción de un orden legal que, entre otros pilares, se basó en la concepción privada de la propiedad de la tierra y la apropiación de riqueza. Orientando nuestra mirada hacia el sur de la provincia de Buenos Aires, a lo largo del siglo XIX, lo que vamos a ver es un proceso de expansión de la frontera, con poblamiento, ocupación, distribución e incorporación de las nuevas tierras al proceso productivo. Las formas que este proceso adquirió constituyen el objeto del presente trabajo

  17. Sexing birds using random amplified polymorphic DNA (RAPD) markers

    NARCIS (Netherlands)

    Lessells, C.M.; Mateman, A.C.

    1998-01-01

    We used random amplified polymorphic DNA (RAPD) markers to sex birds from small tissue (usually blood) samples. Arbitrarily chosen 10-mer PCR primers were screened with DNA from known-sex individuals for the production of a bright female-specific band. Suitable primers were found for seven bird spec

  18. High levels of C-reactive protein in the peripheral blood during visceral leishmaniasis predict subsequent development of post kala-azar dermal leishmaniasis

    DEFF Research Database (Denmark)

    Gasim, S; Theander, T G; ElHassan, A M

    2000-01-01

    Post kala-azar dermal leishmaniasis (PKDL) is a known sequel to visceral leishmaniasis in India and East Africa, and in Sudan about 50% of the kala-azar patients develop PKDL. In this study we followed kala-azar patients from diagnosis and up to 2 years after initiation of treatment. During...... and in keratinocytes during visceral leishmaniasis predict subsequent development of PKDL. The method however requires expensive equipment and reagents. The results of the present study indicate that kala-azar patients, who have a high risk of developing PKDL after treatment can be identified by measuring plasma CRP....

  19. Interest in paromomycin for the treatment of visceral leishmaniasis (kala-azar

    Directory of Open Access Journals (Sweden)

    Wiwanitkit V

    2012-06-01

    Full Text Available Viroj Wiwanitkit1–31Wiwanitkit House, Bang Khae, Bangkok, Thailand; 2Hainan Medical University, Haikou, Hainan, People's Republic of China; 3Joseph Ayo Babalola University, Ikeji-Arakeji, Osun State, NigeriaAbstract: Leishmaniasis is an important vector-borne disease, and it is classified as one of the most important tropical fly-borne infections. This disease can cause two types of clinical manifestations: cutaneous forms and visceral forms. Visceral leishmaniasis, which is also called kala-azar, is a very serious infection that can be fatal. The management of visceral leishmaniasis requires informed diagnostic and therapeutic approaches. Continuous research and development regarding the treatment of visceral leishmaniasis had led to many improvements. Paromomycin is a relatively new antibiotic drug that has been used for the treatment of visceral leishmaniasis for several years. This article reviews and discusses the use of paromomycin for visceral leishmaniasis therapy.Keywords: visceral, leishmaniasis, paromomycin

  20. Relative abundance ofPhlebotominae sandflies with emphasis on vectors of kala-azar

    Institute of Scientific and Technical Information of China (English)

    Naveen Samuel Singh; Doris Phillips-Singh

    2010-01-01

    Objective:To identify potential vectors of kala-azar from Northern plains of Uttar Pradesh, India.Methods:The collections were made using sticky paper traps, CDC light-traps, and aspirators from outdoors as well as indoors. All female sandflies were dissected and identified. Results:During January 2009 to July 2009, 528 phlebotomine specimens were collected including 329 males (62.3%) and 199 females (37.7%), approximately 238 (45%) of them were captured from indoor and 290 (55%) from outdoor resting places. Five species belonging to the genera Phlebotomus (8%) andSergentomyia (92%) were recorded.Conclusions:Public health measures such as case detection and treatment, the control of sandflies, the conjunction elimination of infected stray dogs and health education can be effective in controlling the disease.

  1. Identification of different oyster species using RAPD-PCR

    Directory of Open Access Journals (Sweden)

    Emel Özcan Gökçek

    2017-03-01

    Full Text Available Rapid determination of the genetic differences between some oyster samples observed in the Aegean Sea that were assumed as an invasive species and domestic oyster (Ostrea edulis, Linnaeus 1758 using RAPD (Random Amplified Polymorphic DNA- PCR (Polymerase Chain Reaction markers was aimed in this study. In this regard, domestic species samples and the other samples collected from Altınoluk coasts which were morphologically different than Ostrea species and were showing similarities to Crassostrea genus were genetically indetified by using 8 RAPD profiles. Total 343 bands were observed in the study. Polymorphism rate was found higher in the samples that might belong to Crassostrea genus. 6 of all loci considered in this study were found highly identical for these species. Total of 16 species-spesific diagnostic bands: 11 bands for the invasive species and 5 bands for the domestic species, were determined.

  2. PHYLOGENETIC RELATIONSHIPS AMONG PUFFERFISH OF GENUS TAKIFUGU BY RAPD ANALYSIS

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Amplifications with 300 10-base primers under predetermined optimal reaction yielded 2762 reproducible amplified fragments ranging from 200 to 3000 bp. Genetic distances among five species of Takifugu and Lagocephalus spadiceus as outgroup were calculated based on the presence/absence of the amplified fragments. Topological phylogenic trees of the 5 species of Takifugu and the outgroup were generated by Parsimony and Neighbor-Joining analysis based on RAPD data set. The genetic distance between T. rubripes and T. pseudommus was almost the same as that between individuals within each species, but was much smaller than that among T. rubripes, T. pseudommus and the other species. The molecular data from RAPD analysis convincingly showed that T. rubripes and T. pseudommus should be the same species.

  3. PHYLOGENETIC RELATIONSHIPS AMONG PUFFERFISH OF GENUS TAKIFUGU BY RAPD ANALYSIS

    Institute of Scientific and Technical Information of China (English)

    宋林生; 刘保忠; 王在照; 李红蕾; 相建海; 钱培元

    2001-01-01

    AmpLifications with 300 10-base primers under predetermined optimal reaction yielded 2762 reproducible amplified fragments ranging from 200 to 3000 bp. Genetic distances among five species of Takifugu and Lagocephalus spadiceus as outgreup were calculated based on the presence/absence of the amplified fragments. Topological phylogenie trees of the 5 species of Takgfugu and the outgroup were gen-erated by Parsimony and Neighbor-Joining analysis based on RAPD data set. The genetic distance between T. rubripes and T. pseudommus was almost the same as that between individuals within each species, but was much smaller than that among T. rubripes, T. pseudommus and the other species. The molecu-lar data from RAPD analysis convincingly showed that T. rubripes and T. pseudommus should be the same species.

  4. Selection processes in a citrus hybrid population using RAPD markers

    Directory of Open Access Journals (Sweden)

    Oliveira Roberto Pedroso de

    2003-01-01

    Full Text Available The objective of this work was to evaluate the processes of selection in a citrus hybrid population using segregation analysis of RAPD markers. The segregation of 123 RAPD markers between 'Cravo' mandarin (Citrus reticulata Blanco and 'Pêra' sweet orange (C. sinensis (L. Osbeck was analysed in a F1 progeny of 94 hybrids. Genetic composition, diversity, heterozygosity, differences in chromosomal structure and the presence of deleterious recessive genes are discussed based on the segregation ratios obtained. A high percentage of markers had a skeweness of the 1:1 expected segregation ratio in the F1 population. Many markers showed a 3:1 segregation ratio in both varieties and 1:3 in 'Pêra' sweet orange, probably due to directional selection processes. The distribution analysis of the frequencies of the segregant markers in a hybrid population is a simple method which allows a better understanding of the genetics of citrus group.

  5. RAPD-based genetic relationships in different Bougainvillea cultivars

    Directory of Open Access Journals (Sweden)

    Richa Srivastava

    2009-01-01

    Full Text Available The present study deals with authenticating existing knowledge about 21 Bougainvillea cultivars comprisingof 9 hybrids and their parents through RAPD analysis. The 19 degenerate primer sets generated 234 bands from which 158(67.5% were polymorphic. The UPGMA based dendrogram divided 21 cultivars into two major groups with Jaccard’ssimilarity coefficient ranging from 0.51 to 0.942. Group A had three cultivars namely Trinidad, Formosa and Dr. H. B. Singhin which Dr. H.B. Singh was confirmed as a hybrid of other two cultivars. Group B was sub divided into 8 clusters. Theparentages of 7 out of 8 hybirds have been confirmed based on clusters. The study concluded that the RAPD technique issuitable for confirmation of parent-hybrid relationship.

  6. RAPD analysis of Nectomys squamipes (Rodentia, Sigmodontinae populations

    Directory of Open Access Journals (Sweden)

    Almeida Francisca C.

    2000-01-01

    Full Text Available Random amplified of polymorphic DNA (RAPD analysis was used to assess genetic distance and the genetic structure of populations of Nectomys squamipes, a semiaquatic rodent species distributed along watercourses. DNA samples of five populations were analyzed using three primers, producing 45 scorable bands, 31 of which were polymorphic. There was a significant differentiation among populations [F ST = 0.17; phiST = 0.14 (P < 0.004] but gene flow (Nm = 1.25 was sufficient to overcome genetic drift effects. No fixed specific markers were found for any population. The Mantel's test and UPGMA cluster analysis showed a lack of relationship between genetic and geographic distances. The apparent homogeneity indicated by RAPD markers coincided with morphometric data, despite the wide geographic range of N. squamipes. Alternative hypotheses for explaining our results include recurrent processes of local extinction and recolonization or a recent and sudden increase in the geographic distribution of this species.

  7. RAPD markers related to sex locus in Populus tomentosa

    Institute of Scientific and Technical Information of China (English)

    Wanwei HOU; Junfeng FAN; Feimei ZHOU; Shufang ZHAO

    2009-01-01

    By using the methods of random amplified polymorphic DNA (RAPD) and bulked segregate analysis (BSA), we identified markers that are linked to the sex determination in the dioecious Populus tomentosa. Male and female bulks were created through rough mixing equal amounts of its five individual DNA. A total of 88 primers were screened. Twelve primers produced clear patterns with at least one band that appeared to be polymorphic between the two bulks. Subsequently, five male and female individuals were analyzed with those 12 primers, and only S60 (ACCCGGTCAC) could generate a common 1800bp DNA fragment in all five male individuals and male pool but not in any female individuals. It can be concluded that the gender of P. tomentosa is most likely connected to the S60-1800bp DNA fragment and RAPD markers. S60, therefore, can be used for selecting the gender of P. tomentosa.

  8. Screening and characterization of RAPD markers in viscerotropic Leishmania parasites.

    Directory of Open Access Journals (Sweden)

    Imen Mkada-Driss

    Full Text Available Visceral leishmaniasis (VL is mainly due to the Leishmania donovani complex. VL is endemic in many countries worldwide including East Africa and the Mediterranean region where the epidemiology is complex. Taxonomy of these pathogens is under controversy but there is a correlation between their genetic diversity and geographical origin. With steady increase in genome knowledge, RAPD is still a useful approach to identify and characterize novel DNA markers. Our aim was to identify and characterize polymorphic DNA markers in VL Leishmania parasites in diverse geographic regions using RAPD in order to constitute a pool of PCR targets having the potential to differentiate among the VL parasites. 100 different oligonucleotide decamers having arbitrary DNA sequences were screened for reproducible amplification and a selection of 28 was used to amplify DNA from 12 L. donovani, L. archibaldi and L. infantum strains having diverse origins. A total of 155 bands were amplified of which 60.65% appeared polymorphic. 7 out of 28 primers provided monomorphic patterns. Phenetic analysis allowed clustering the parasites according to their geographical origin. Differentially amplified bands were selected, among them 22 RAPD products were successfully cloned and sequenced. Bioinformatic analysis allowed mapping of the markers and sequences and priming sites analysis. This study was complemented with Southern-blot to confirm assignment of markers to the kDNA. The bioinformatic analysis identified 16 nuclear and 3 minicircle markers. Analysis of these markers highlighted polymorphisms at RAPD priming sites with mainly 5' end transversions, and presence of inter- and intra- taxonomic complex sequence and microsatellites variations; a bias in transitions over transversions and indels between the different sequences compared is observed, which is however less marked between L. infantum and L. donovani. The study delivers a pool of well-documented polymorphic DNA markers

  9. How to test for a relative afferent pupillary defect (RAPD

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    David C Broadway

    2012-01-01

    Full Text Available The 'swinging light test' is used to detect a relative afferent pupil defect (RAPD: a means of detecting differences between the two eyes in how they respond to a light shone in one eye at a time. The test can be very useful for detecting unilateral or asymmetrical disease of the retina or optic nerve (but only optic nerve disease that occurs in front of the optic chiasm.

  10. RAPD analysis of natural populations of Acanthopanax brachypus

    Institute of Scientific and Technical Information of China (English)

    YANHUAJUN; SILANDAI; 等

    1997-01-01

    Random amplified Polymorphic DNA(RAPD) analysis is a new technology of molecular marking which has proved very powerful in detecting genetic diversity at the level of population.The genomic DNAs used in our experiment were extracted from fresh leaves taken from 59 individuals sampled from three natural populations in Yan an,Shanxi Province.Through more than 2,000 PCRs,deep-going RAPD analysis was carried out on DNA samples from 49 inviduals.The percentage of polymorphic RAPD loci found in these three populations were respectively 27.2%,18.6% and 5.4%;the average genetic distances within population,0.055,0.026 and 0.008;the average genetic distances between populations (I-II),(I-III) and (II-III),0.105,0.096 and 0.060.The genetic diversity of A.brachypus within and between populations was found,for the first time,to be rather poor,thus revealing innate factors as the cause contributing to its endangered status.In addition,our work also provides basic materials for elucidating the underlying cause of its endangerment and for its protection biology.

  11. The use of RAPD to characterize Bipolaris sorokiniana isolates.

    Science.gov (United States)

    Müller, Maria Viviane Gomes; Germani, José Carlos; Van Der Sand, Sueli Teresinha

    2005-01-01

    Bipolaris sorokiniana is a phytopathogenic fungus causing diseases of cereal crops such as common root rot, the leaf spot disease, seedling blight, and black point of the grain. Random-amplified polymorphic DNA (RAPD) assay was used to investigate the genetic diversity of 20 isolates collected from different cultivars in wheat-producing regions in Brazil. Seventy primers, with random nucleotide sequences, were tested. Reproducibility to amplify the genomic DNA of isolates was found for 30 of the 70 primers tested, generating between 1 and 17 fragments ranging from 0.35 to 2.0 kb (average size). The degree of similarity between samples was calculated through simple association and the dendrogram was assessed using the unweighted pair group method with arithmetical average. After the RAPD analyses 19 isolates were closely grouped, having a similarity coefficient of >or= 78%. Isolate I017 showed very low similarity coefficients, ranging between 38 and 46%. The RAPD analyses provided important information as to the degree of genetic variability and the relationship between the isolates investigated, revealing polymorphism and establishing electrophoretic profiles useful to characterize the phytopathogen.

  12. Genetic relationships among Heliconia (Heliconiaceae) species based on RAPD markers.

    Science.gov (United States)

    Marouelli, L P; Inglis, P W; Ferreira, M A; Buso, G S C

    2010-07-13

    The family Heliconiaceae contains a single genus, Heliconia, with approximately 180 species of Neotropical origin. This genus was formerly allocated to the family Musaceae, but today forms its own family, in the order Zingiberales. The combination of inverted flowers, a single staminode and drupe fruits is an exclusive characteristic of Heliconia. Heliconias are cultivated as ornamental garden plants, and are of increasing importance as cut flowers. However, there are taxonomic confusions and uncertainties about the number of species and the relationships among them. Molecular studies are therefore necessary for better understanding of the species boundaries of these plants. We examined the genetic variability and the phylogenetic relationships of 124 accessions of the genus Heliconia based on RAPD markers. Phenetic and cladistic analyses, using 231 polymorphic RAPD markers, demonstrated that the genus Heliconia is monophyletic. Groupings corresponding to currently recognized species and some subgenera were found, and cultivars and hybrids were found to cluster with their parents. RAPD analysis generally agreed with morphological species classification, except for the position of the subgenus Stenochlamys, which was found to be polyphyletic.

  13. Genetic variation in Phoca vitulina (the harbour seal) revealed by DNA fingerprinting and RAPDs

    NARCIS (Netherlands)

    Kappe, A.L.; van de Zande, L.; Vedder, E.J.; Bijlsma, R.; van Delden, Wilke

    1995-01-01

    Genetic variation in two harbour seal (Phoca vitulina) populations from the Dutch Wadden Sea and Scotland was examined by RAPD analysis and DNA fingerprinting. For comparison a population of grey seals (Halichoerus grypus) was studied. The RAPD method revealed a very low number of polymorphic bands.

  14. Selection of unique Escherichia coli clones by random amplified polymorphic DNA (RAPD)

    DEFF Research Database (Denmark)

    Nielsen, Karen L; Godfrey, Paul A; Stegger, Marc

    2014-01-01

    Identifying and characterizing clonal diversity are important when analysing fecal flora. We evaluated random amplified polymorphic DNA (RAPD) PCR, applied for selection of Escherichia coli isolates, by whole genome sequencing. RAPD was fast, and reproducible as screening method for selection of ...

  15. Assessing the limits of random amplified polymorphic DNAs (RAPDs) in seaweed biogeography

    NARCIS (Netherlands)

    vanOppen, MJH; Klerk, H; deGraaf, M; Stam, WT; Olsen, JL

    1996-01-01

    As judged by comparison with other molecular data sets, random amplified polymorphic DNA (RAPD) data are robust in identifying large-scale biogeographic populations that range from hundreds to thousands of kilometers apart. As the geographical scale is shifted downward, however, RAPD data often fail

  16. Direct RAPD evaluation of bacteria without conventional DNA extraction

    Directory of Open Access Journals (Sweden)

    Welington Luiz Araújo

    2004-07-01

    Full Text Available The present work reports successful DNA amplification of Pantoea agglomerans and Bacillus pumilus through Random Amplified Polymorphic DNA (RAPD. For this, template DNA was obtained without conventional DNA extraction. The procedure was as follows: cultures grown for 20 hours in 5 mL LB medium were centrifuged and the resulting preparation was suspended in TE buffer. After boiling, the cell suspension was diluted and 2.0 µl were used in reactions of 15 µl. The results showed no significant differences among the RAPD profile of the PCR reactions derived from the boiling and phenol extraction methods, suggesting the utilization of this method for genetic population analysis.O presente trabalho mostra a amplificação de DNA das bactérias Pantoea agglomerans e Bacillus pumilus por meio da técnica de RAPD (Amplificação ao acaso de DNA polimórfico. Para esta análise, o DNA molde foi obtido sem a utilização de técnicas de extração convencional, ou seja, sem a purificação do DNA. Bactérias foram cultivadas por 20 horas em 5 mL de meio LB, centrifugado e ressuspendido em tampão TE. A suspensão resultante foi fervida por 5 min., diluída e 2,0 µL foram usados em reações de 15 µL. Os resultados mostraram que os padrões observados com o DNA obtido pela fervura das células não apresentou diferenças significativas daquele obtido com DNA extraído e purificado com fenol, sugerindo a possibilidade da utilização deste método para o estudo da variabilidade genética de populações microbianas.

  17. Reevaluation of RAPD markers involved in a case of stingray misidentification (Dasyatidae: Dasyatis).

    Science.gov (United States)

    Faria, V V; Rolim, L S; Vaz, L A L; Furtado-Neto, M A A

    2012-10-25

    We investigated a reported case of stingray Dasyatis americana misidentification not detected in a published study using the random amplified polymorphic DNA (RAPD) technique. If the referred specimen (landed by fisheries in Ceará, northeastern Brazil) was misidentified (as Dasyatis centroura) in the field, why did its RAPD data fail to clarify the mistake? Was it due to limitations of RAPD markers or perhaps to a taxonomic issue? Contrary to our initial expectations, neither of these hindered the detection of the misidentification. After reanalyzing the primary genetic data associated with the misidentified specimen (PCR gel photographs and/or matrices of presence/absence of markers for six RAPD primers), we found that the RAPD markers were sufficient to correctly assign the misidentified specimen to its proper species identity. In the original study, the specimen misidentification was neither noticed by the authors nor apparent in the published article due to how their results were interpreted and presented.

  18. XML Prevalence of Klebsiella Pneumoniae in Panje Azar Hospital of Gorgan, Iran, 2011

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    Ma hmoudjanlou, H. (MSc

    2015-01-01

    Full Text Available Background and Objective: Klebsiella pneumoniae is one of the agents causing nosocomial infection; therefore, we decided to report the prevalence of Klebsiella pneumoniae caused infection. Material and methods: The frequency of Klebsiella in culture media samples of Panje Azar hospital was studied in 2011-2012. After determination of the species with biochemical methods and determination of resistance to third generation cephalosporins, the existence of responsible genes for this resistance was investigated using specific primers. The PCR product for CTX-M gene was sequenced. Results: During the study, 70 isolates of Klebsiella were isolated in that 51 (72.8% related to three months of November, December and January. Except for the one related to November, other ESBL cases belonged to these three months. Based on molecular investigation of ESBL genes, these isolates at least were in 3 types and had a high frequency in Internal, female and Emergency wards. Conclusion: The present report implied a sudden prevalence of Klebsiella pneumoniae that detected and controlled by a correct monitoring.

  19. Caracterização molecular de butiazeiro por marcadores RAPD Molecular characterization of Pindo palm by RAPD markers

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    Adrise Medeiros Nunes

    2008-09-01

    Full Text Available O grupo botânico Arecaceae é de extremo interesse por compreender plantas em extinção e por apresentar um grande potencial de exploração econômica. O butiazeiro (Butia capitata (Mart. Becc. ocorre naturalmente no Sul do Brasil. Sua caracterização molecular é de extremo interesse para futuros trabalhos de melhoramento genético. Assim sendo, verificou-se a variabilidade genética existente entre vinte e dois genótipos de butiazeiro da espécie (Butia capitata, pertencentes ao BAG (Banco Ativo de Germoplasma de frutíferas nativas do Centro Agropecuário da Palma - UFPel. Esses genótipos foram analisados usando marcadores do tipo RAPD (Random Amplified Polymorphic DNA. Um total de 136 fragmentos foram obtidos, sendo 77 polimórficos. O primer OPA11 apresentou maior polimorfismo, produzindo 9 perfis diferentes. A análise de agrupamento, realizada pelo método UPGMA, produziu um dendrograma que permitiu a clara separação dos genótipos em dois grupos principais. Verificou-se que, com a técnica de marcadores de RAPD, foi possível obter um perfil molecular único e uma estimativa da variabilidade existente entre os genótipos de butiazeiro avaliados.The study of the botanical group Arecaceae is of extreme interest for evolving several endangered species of plants and for presenting a great potential of economical exploration. The Pindo palm (or wine palm, jelly palm (Butia capitata (Mart. Becc. is natural from the south of Brazil. Its molecular characterization is of extreme interest for future researches of genetic improvement. Since little is known about the variability of the species, the existent genetic variability was verified among twenty-two genotypes of Pindo palm (or wine palm, jelly palm, from BAG (Germoplasm Assets Bank of fruit trees native from the Agricultural Center of the Palma - UFPEL, which were analyzed using markers RAPD (Random Amplified Polymorphic DNA with Operon Technologies' decamers primers. With 21 primers

  20. Genetic relationships among Orobanche species as revealed by RAPD analysis.

    Science.gov (United States)

    Román, B; Alfaro, C; Torres, A M; Moreno, M T; Satovic, Z; Pujadas, A; Rubiales, D

    2003-05-01

    RAPD markers were used to study variation among 20 taxa in the genus OROBANCHE: O. alba, O. amethystea, O. arenaria, O. ballotae, O. cernua, O. clausonis, O. cumana, O. crenata, O. densiflora, O. foetida, O. foetida var. broteri, O. gracilis, O. haenseleri, O. hederae, O. latisquama, O. mutelii, O. nana, O. ramosa, O. rapum-genistae and O. santolinae. A total of 202 amplification products generated with five arbitrary RAPD primers was obtained and species-specific markers were identified. The estimated Jaccard's differences between the species varied between 0 and 0.864. The pattern of interspecific variation obtained is in general agreement with previous taxonomic studies based on morphology, and the partition into two different sections (Trionychon and Orobanche) is generally clear. However, the position in the dendrogram of O. clausonis did not fit this classification since it clustered with members of section TRIONYCHON: Within this section, O. arenaria was relatively isolated from the other members of the section: O. mutelii, O. nana and O. ramosa. Within section Orobanche, all O. ramosa populations showed a similar amplification pattern, whereas differences among O. crenata populations growing on different hosts were found. Orobanche foetida and O. densiflora clustered together, supporting the morphological and cytological similarities and the host preferences of these species.

  1. Genetic diversity in Hemileia vastatrix based on RAPD markers.

    Science.gov (United States)

    Gouveia, M Manuela C; Ribeiro, Ana; Várzea, Vítor M P; Rodrigues, Carlos J

    2005-01-01

    Random amplified polymorphic DNA (RAPD) was used to assess the genetic structure of Hemileia vastatrix populations. Forty-five rust isolates with different virulence spectra and from different hosts and geographical regions were analyzed. Out of 45 bands, generated with three RAPD primers, 35 (78%) were polymorphic and scored as molecular markers. Cluster analysis exhibits unstructured variability of this pathogen with regard to physiological race, geographical origin or host. The genotypic diversity (H') inferred from Shannon's index was higher than gene diversity (Ht), suggesting that diversity is distributed among clonal lineages. Estimates of gene diversity in Africa and Asia populations were higher in total (Ht) as compared to within population diversity (Hs). Genetic differentiation was considerable among coffee rust isolates from Africa (Gst = 0.865) and Asia (Gst = 0.768) but not among isolates from South America (Gst = 0.266). We concluded that genetic diversity in H. vastatrix was moderately low and that the genetic differentiation among populations shows that asexual reproduction is likely to play an important role in the population biology of this fungus. This should be taken into account for the development of breeding programs.

  2. Characterization of rhizobia that nodulate Arachis pintoi by RAPD analysis Caracterização de rizóbios capazes de nodular Arachis pintoi via análise de "RAPD"

    Directory of Open Access Journals (Sweden)

    Patrícia Pereira Pinto

    2004-09-01

    Full Text Available The genetic relationships of 85 Arachis pintoi nodulating Rhizobium strains were determined using the random amplified polymorphic DNA (RAPD methods. The analysis included 75 strains isolated from Cerrado soils and 10 other ones of different origins. The results indicated that there is a high level of similarity between these strains and that geographic distribution may affect their phylogenetic relationship. In addition, the results allowed the selection of the most suitable primers for characterisation of these Rhizobium strains which will be useful for implementation of competitiveness studies in Cerrado soils.As relações genéticas de 85 estirpes de Rhizobium capazes de nodular Arachis pintoi foram determinadas usando o método de "RAPD" (Random Amplified Polymorphic DNA. As análises incluíram 75 estirpes isoladas de solos de Cerrado e 10 de diferentes origens. Os resultados indicaram que existe um alto grau de similaridade entre estas estirpes e que a distribuição geográfica pode afetar suas relações filogenéticas. Além disso, os resultados permitiram a seleção de "primers" mais adequados para a caracterização dessas estirpes de Rhizobium, os quais serão úteis para a implementação de estudos de competitividade nos solos de Cerrado.

  3. Asymptomatic leishmaniasis in kala-azar endemic areas of Malda district, West Bengal, India

    Science.gov (United States)

    Saha, Pabitra; Ganguly, Swagata; Chatterjee, Moytrey; Das, Soumendu Bikash; Kundu, Pratip K.; Guha, Subhasish K.; Ghosh, Tamal K.; Bera, Dilip K.; Basu, Nandita

    2017-01-01

    Asymptomatic leishmaniasis may drive the epidemic and an important challenge to reach the goal of joint Visceral Leishmaniasis (VL) elimination initiative taken by three Asian countries. The role of these asymptomatic carriers in disease transmission, prognosis at individual level and rate of transformation to symptomatic VL/Post Kala-azar Dermal Leishmaniasis (PKDL) needs to be evaluated. Asymptomatic cases were diagnosed by active mass survey in eight tribal villages by detecting antileishmanial antibody using rK39 based rapid diagnostic kits and followed up for three years to observe the pattern of sero-conversion and disease transformation. Out of 2890 total population, 2603 were screened. Antileishmanial antibody was detected in 185 individuals of them 96 had a history of VL/PKDL and 89 without such history. Seventy nine such individuals were classified as asymptomatic leishmaniasis and ten as active VL with a ratio of 7.9:1. Out of 79 asymptomatic cases 2 were lost to follow up as they moved to other places. Amongst asymptomatically infected persons, disease transformation in 8/77 (10.39%) and sero-conversion in 62/77 (80.52%) cases were noted. Seven (9.09%) remained sero-positive even after three years. Progression to clinical disease among asymptomatic individuals was taking place at any time up to three years after the baseline survey. If there are no VL /PKDL cases for two or more years, it does not mean that the area is free from leishmaniasis as symptomatic VL or PKDL may appear even after three years, if there are such asymptomatic cases. So, asymptomatic infected individuals need much attention for VL elimination programme that has been initiated by three adjoining endemic countries. PMID:28187202

  4. Entre la ley y el azar: La trama vincular del mundo político-empresarialde la frontera sur pampeana en el siglo XIX

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    Andrea Reguera

    2006-12-01

    Full Text Available Entre la ley y el azar hace referencia a un momento muy particular de nuestra historia argentina, el de la construcción de un orden legal que, entre otros pilares, se basó en la concepción privada de la propiedad de la tierra y la apropiación de riqueza. Orientando nuestra mirada hacia el sur de la provincia de Buenos Aires, a lo largo del siglo XIX, lo que vamos a ver es un proceso de expansión de la frontera, con poblamiento, ocupación, distribución e incorporación de las nuevas tierras al proceso productivo. Las formas que este proceso adquirió constituyen el objeto del presente trabajoBetween the Law and the Hazards refers to a very particular moment of Argentine history. A period where the construction of law and order was build upon a conception of private property of the land as well as private appropriation of wealth. We turn our view towards the southern region of Buenos Aires province all through the 19th century, in order to observe a process of expansion of the frontier line which included the settlement, occupation, distribution and incorporation of new lands to the productive process. It is the aim of our work to study the forms this process actually took

  5. Development of RAPD-SCAR markers for different Ganoderma species authentication by improved RAPD amplification and molecular cloning.

    Science.gov (United States)

    Fu, J J; Mei, Z Q; Tania, M; Yang, L Q; Cheng, J L; Khan, M A

    2015-05-25

    The sequence-characterized amplified region (SCAR) is a valuable molecular technique for the genetic identification of any species. This method is mainly derived from the molecular cloning of the amplified DNA fragments achieved from the random amplified polymorphic DNA (RAPD). In this study, we collected DNA from 10 species of Ganoderma mushroom and amplified the DNA using an improved RAPD technique. The amplified fragments were then cloned into a T-vector, and positive clones were screened, indentified, and sequenced for the development of SCAR markers. After designing PCR primers and optimizing PCR conditions, 4 SCAR markers, named LZ1-4, LZ2-2, LZ8-2, and LZ9-15, were developed, which were specific to Ganoderma gibbosum (LZ1-4 and LZ8-2), Ganoderma sinense (LZ2-2 and LZ8-2), Ganoderma tropicum (LZ8-2), and Ganoderma lucidum HG (LZ9-15). These 4 novel SCAR markers were deposited into GenBank with the accession Nos. KM391935, KM391936, KM391937, and KM391938, respectively. Thus, in this study we developed specific SCAR markers for the identification and authentication of different Ganoderma species.

  6. Genetic divergence of Scots pine (Pinus sylvestris L. populations in Serbia revealed by RAPD

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    Lučić A.

    2011-01-01

    Full Text Available The ability of random amplified polymorphic DNA (RAPD to distinguish among Scots pine populations from Serbia was evaluated. Sixteen arbitrary 10-mer primers employed in the analysis produced 54 fragments of which 21 were polymorphic (38.89%. Certain rare and genotype-specific bands were identified which could be effectively used to distinguish between the populations. Polymorphism in RAPD markers among P. sylvestris populations was high and sufficient to distinguish each of the populations. The results obtained suggest that RAPD markers are valuable for the genetic divergence estimation in Pinus sylvestris and for the study of divergence among populations.

  7. Increased natural killer activity does not prevent progression of experimental Kala-azar Atividade "natural-killer" aumentada não impede a progressão do Kala-azar experimental

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    Alexandrina SARTORI

    1999-07-01

    Full Text Available Kala-azar is the visceral form of leishmaniasis and it is caused by intracellular parasites from the complex Leishmania donovani. Golden hamster (Mesocricetus auratus infected with Leishmania donovani develop a disease very similar to human Kala-azar. There is conspicuous hipergammaglobulinaemia and their T cells do not respond to stimulation with parasite antigens. We used this experimental model to evaluate the natural killer (NK activity during the initial phase of the disease. Outbred hamsters infected by intravenous route with 5.106 amastigotes of L. donovani 1S showed a concurrent increase in the spleen weight and in the spleen cell number. Using the single cell assay we detected a significant increase in the percentage of NK effector cells on the 4th day of infection. Imprints from spleen and liver showed at days 14 and 28 a significant increase in the parasite burden . These results show that the increased NK activity in the beginning of the infection was not able to restrain the progression of the disease in this experimental model.O Kala-azar é a forma visceral da leishmaniose e é causado pelos parasitas do complexo Leishmania donovani. O hamster dourado (Mesocricetus auratus infectado com L. donovani desenvolve uma doença bastante similar ao Kala-azar humano, apresentando hipergamaglobulinemia e supressão da resposta imune celular específica. Utilizamos este modelo experimental para avaliar a atividade natural killer (NK na fase inicial da infecção. Hamsters não isogênicos infectados por via intravenosa com 5.106 amastigotas de L. donovani 1S apresentaram aumento no peso do baço e no número de células esplênicas. Utilizando o "single cell assay" detectamos um aumento significativo no percentual de células NK efetoras no 4º dia de infecção. "Imprints" de baço e de fígado mostraram aumento significativo na carga parasitária após 14 e 28 dias de infecção. Os resultados mostram que o aumento da atividade NK

  8. Identification of common bean alleles resistant to anthracnose using RAPD

    Directory of Open Access Journals (Sweden)

    Ana L.M. Castanheira

    1999-12-01

    Full Text Available RAPD markers were identified close to common bean alleles responsible for resistance to the fungus Colletotrichum lindemuthianum and may be useful in selecting plants resistant to this pathogen. DNA from F2 plants of the crosses Carioca 300V x P45, Carioca 300V x Ouro and P24 x Ouro was amplified by RAPD. Line P45 has the Co.4 allele for resistance, and the Ouro cultivar has the Co.5 allele. The primer OPC08 amplified a DNA fragment of about 1059 bp linked to the Co.4 allele. The recombination frequency was 0.133 (SE = 0.039; 95% CI = 0.056-0.211. Using the primer OPF10 a DNA fragment of about 912 bp was amplified and found to be associated with the Co.5 allele. The recombination frequency was 0.115 (SE = 0.038; 95% CI = 0.041-0.189. A second marker (1122 pb amplified by the OPR03 primer was identified in the population P24 x Ouro. The recombination frequency for this marker was 0.363 (SE = 0.081; 95% CI = 0.205-0.522. Both these markers flanked the Co.5 allele. The markers identified in this study may be useful in identifying lines with the Co.4 and Co.5 alleles.Marcadores RAPD foram identificados próximos de alelos do feijão responsáveis pela resistência ao Colletotrichum lindemuthianum, visando auxiliar na seleção de plantas resistentes ao patógeno. Empregou-se o método dos bulks segregantes de DNA extraídos de plantas F2 dos seguintes cruzamentos: Carioca 300V x P45, Carioca 300V x Ouro e P24 x Ouro. A linhagem P45 é portadora do alelo Co.4 de resistência e o cultivar Ouro é portador do alelo Co.5, os quais foram marcados. Procedeu-se à reação RAPD dos bulks e foi identificado o iniciador OPC08 que amplificou um fragmento de DNA com cerca de 1059 pb, ligado ao alelo Co.4. A freqüência de recombinação foi de 0,133 (erro padrão 0,039 e o intervalo de confiança foi 0,056 e 0,211, com 95% de probabilidade. Em relação ao alelo Co.5 foi identificado um fragmento de DNA amplificado pelo iniciador OPF10 com cerca de 912 pb, na

  9. Invasion and spreading of Cabomba caroliniana revealed by RAPD markers

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    Applying randomly amplified polymorphic DNA (RAPD), the genetic variation of Cabomba caroliniana Gray (cabomba or fanwort), a new alien plant in China, was analyzed in this paper. Total 143 bands, including 47 polymorphic bands, were amplified from 23 primers in 20 samples. The sampling distance was large,but its genetic diversity was low. The main results were that: (1) Cabomba, which grew and dispersed mainly in fragment, was an abundant and dominant species in freshwater, and its main dispersal mechanism was vegetative reproduction. (2) Cabomba was originally introduced into China as an aquarium submerged plant. Somehow,those discarded cabomba became invasive species in the areas of Hangzhou, Shanghai, and Meicheng, and other places. (3) Although the level of genetic diversity in cabomba was low, their rapid dispersion and propagation could seriously harm to local aquatic community. Therefore, specific measure should be used to control cabomba from uncontrolled spreading and damage to local vegetation communities.

  10. Genetic Diversity of RAPD Mark for Natural Davidia involucrata Populations

    Institute of Scientific and Technical Information of China (English)

    Congwen Song; Manzhu Bao

    2006-01-01

    The genetic diversity and genetic variation within and among populations of five natural Davidia involucrata populations were studied from 13 primers based on random amplified polymorphic DNA (RAPD) analysis.The results show that natural D.involucrata population has a rich genetic diversity,and the differences among populations are significant.Twenty-six percent of genetic variation exists among D.involucrata populations,which is similar to that of the endangered tree species Liriodendron chinense and Cathaya argyrophylla in China,but different from more widely distributed tree species.The analysis of the impacts of sampling method on genetic diversity parameters shows that the number of sampled individuals has little effect on the effective number of alleles and genetic diversity,but has a marked effect on the genetic differentiation among populations and gene flows.This study divides the provenances of D.involucrata into two parts,namely,a southeast and a northwest provenance.

  11. RAPD and phytochemical analysis of Thymus moroderi plantlets after cryopreservation.

    Science.gov (United States)

    Marco-Medina, Ana; Casas, José Luis

    2013-01-01

    Cryopreservation is at present the most reliable strategy to preserve plant germplasm. When aromatic plants are the object of conservation it is necessary to assess not only the genetic but also the phytochemical stability to ensure that plant material maintains its qualities after storage. In this work we present molecular and phytochemical stability data related to a previously described vitrification-based cryopreservation protocol for Thymus moroderi Pau ex Martínez. RAPD markers have been used to assess the genetic stability of T. moroderi explants and revealed 0.34 percent of variation in the cryopreserved material studied. Phytochemical data collected from GC-MS analysis of dichloromethane extracts from cryopreserved plantlets rendered a profile in which 1,8-cineole (14.5 percent), camphor (5.9 percent) and borneol (5.2 percent) were the major components. Both data confirmed the suitability of the cryopreservation protocol applied.

  12. Alterações cutâneas do cão no Kala-Azar sul-americano

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    C. Magarinos Torres

    1941-01-01

    Full Text Available According to E. Chagas (1938, South-American Kala Azar is a widespread disease from the jungle, several cases being reported from North Brazil (Estado do Pará: Marajó Island, Tocantins and Gurupi river valleys; Estados do Piauí and Ceará: coast and hinterland. Other cases were found in Northeast Brazil (Estados de Pernambuco, Alagôas and Sergipe: coast and hinterland; Estado da Bahia: hinterland. A few cases were described from Estado de Mato-Grosso (Brazil, Provincia de Salta and Território do Chaco (Argentine, and Zona contestada do Chaco (Paraguai-Bolívia. A well defined secondary anemia associated with enlargement of the liver and spleen are the chief symptoms. Death usually occurs in cachexia and with symptoms of heart failure. Half the patients were children aged less than ten years (CHAGAS, CASTRO & FERREIRA, 1937. Quite exhaustive epidemiological researches performed by CHAGAS, FERREIRA, DEANE, DEANE & GUIMARÃES (1938 in Municipio de Abaeté (Estado do Pará, Brazil gave the incidence of 1.48% for the natural infection in human, 4.49% in dogs, and 2.63% in cats. The infection was arcribed (CUNHA & CHAGAS, 1937 to a new species of Leishmania (L. chagasi. Latter CUNHA (1938 state, that it is identical to L. infantum. ADLER (1940 found that so far it has been impossible to distinguish L. chagasi from L. infantum by any laboratory test but a final judgment must be reserved until further experiments with different species of sandflies have been carried out. Skin changes in canine Kala Azar were signaled by many workers, and their importance as regards the transmission of the disease is recognized by some of them (ADLER & THEODOR, 1931, 2. CUNHA, 1933. Cutaneous ulcers in naturally infected dogs are referred by CRITIEN (1911 in Malta, by CHODUKIN & SCHEVTSCHENKO (1928 in Taschkent, by DONATIEN & LESTOCQUARD (1929 and by LESTOCQUARD & PARROT (1929 in Algeria, and by BLANC & CAMINOPETROS (1931 in Greece. Depilation is signaled by

  13. Discrimination of Wild Tea Germplasm Resources (Camellia sp.) Using RAPD Markers

    Institute of Scientific and Technical Information of China (English)

    CHEN Liang; WANG Ping-sheng; Yamaguchi Satoshi

    2002-01-01

    Discrimination of 24 wild tea germplasm resources ( Camellia sp. ) using RAPD markers was conducted. The result showed that RAPD markers were very effective tool and method in wild tea germplasm discrimination. There were 3 independent ways to discriminate tea germplasms, a) unique RAPD markers, b)specific band patterns and c) a combination of the band patterns or DNA fingerprinting provided by different primers. The presence of 16 unique RAPD markers and the absence of 3 unique markers obtained from 12 primers made it possible to discriminate 14 germplasms. Using the unique band patterns of primer OPO-13 could discriminate 10 tea germplasms. It was of much importance using minimum primers to obtain maximum discrimination capacity. All the 24 wild tea germplasms could be discriminated easily and entirely by the band patterns combination or DNA fingerprinting obtained from OPO-13, OPO-18, OPG-12 and OPA-13, including two wild tea trees of very similar morphological characteristics and chemical components.

  14. Genetic divergence of Scots pine (Pinus sylvestris L.) populations in Serbia revealed by RAPD

    OpenAIRE

    Lučić A.; Isajev V.; Rakonjac L.; Ristić Danijela; Kostadinović Marija; Babić Vojka; Nikolić Ana

    2011-01-01

    The ability of random amplified polymorphic DNA (RAPD) to distinguish among Scots pine populations from Serbia was evaluated. Sixteen arbitrary 10-mer primers employed in the analysis produced 54 fragments of which 21 were polymorphic (38.89%). Certain rare and genotype-specific bands were identified which could be effectively used to distinguish between the populations. Polymorphism in RAPD markers among P. sylvestris populations was high and sufficient to distinguish each of the popul...

  15. Isozyme and RAPD studies in Prosopis glandulosa and P. Velutina (Leguminosae, Mimosoideae

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    Bessega Cecilia

    2000-01-01

    Full Text Available Allozyme and random amplified polymorphic DNA (RAPD techniques have been compared for their usefulness for genetic and taxonomic studies in Prosopis glandulosa and P. velutina populations. Isozymes and RAPDs yielded similarly high estimates of genetic variability. Genetic structure and differentiation were analyzed through non-hierarchical Wright's F DT. For all populations considered, both markers produced low gene flow (Nm 1, in agreement with that expected for conspecific populations. However, in RAPD data the expected reduction in F DT and the increase in Nm were not observed. Correlation between F DT and geographical distance matrices (Mantel test for all populations was significant (P = 0.02 when based on isozymes, but not so (P = 0.33 when based on RAPDs. No significant associations among genetic and geographical or climatic variables were observed. Two isoenzyme systems (GOT and PRX enabled us to distinguish between P. glandulosa and P. velutina, but no diagnostic band for recognition of populations or species studied here were detected by RAPD. However, RAPD markers showed higher values for genetic differentiation among conspecific populations of P. glandulosa and a lower coefficient of variation than those obtained from isozymes.

  16. Seasonal relationship between normalized difference vegetation index and abundance of the Phlebotomus kala-azar vector in an endemic focus in Bihar, India

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    Gouri S. Bhunia

    2012-11-01

    Full Text Available Remote sensing was applied for the collection of spatio-temporal data to increase our understanding of the potential distribution of the kala-azar vector Phlebotomus argentipes in endemic areas of the Vaishali district of Bihar, India. We produced monthly distribution maps of the normalized difference vegetation index (NDVI based on data from the thematic mapper (TM sensor onboard the Landsat-5 satellite. Minimum, maximum and mean NDVI values were computed for each month and compared with the concurrent incidence of kala-azar and the vector density. Maximum and mean NDVI values (R2 = 0.55 and R2 = 0.60, respectively, as well as the season likelihood ratio (X2 = 17.51; P <0.001, were found to be strongly associated with kala-azar, while the correlation with between minimum NDVI values and kala-azar was weak (R2 = 0.25. Additionally, a strong association was found between the mean and maximum NDVI values with seasonal vector abundance (R2 = 0.60 and R2 = 0.55, respectively but there was only a marginal association between minimum NDVI value and the spatial distribution of kala-azar vis-à-vis P. argentipes density.

  17. Influence of topography on the endemicity of Kala-azar: a study based on remote sensing and geographical information system

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    Gouri S. Bhunia

    2010-05-01

    Full Text Available Kala-azar, a fatal infectious disease in many Indian states, particularly in Bihar, West Bengal, Uttar Pradesh, and Jharkhand, is caused by the protozoan parasite Leishmania donovani and transmitted by the sandfly vector Phlebotomus argentipes. The vector is distributed all over the country but the disease is confined to particular zones since before the last century. In this study, parameters such as altitude, temperature, humidity, rainfall and the normalized difference vegetation index (NDVI were investigated for correlation with the distribution of the disease in the northeastern corner of the Indian sub-continent. Data analysis on Kala-azar prevalence during the period 2005-2007 in the four states showed that the highest prevalence was below 150 m of altitude with very few cases located above the 300 m level. Low NDVI value ranges (0.03-0.015 correlated with a high occurrence of the disease. The maximum temperatures in the affected sites varied between an upper level of 25-29°C and a minimum of 16-20°C. The rainfall in these areas fluctuated between 1154 and 1834 mm. As the disease showed a high correlation with the prevailing topographic conditions, an attempt was made to improve the relative strength of the approach to predict the potential for endemicity of leishmaniasis by introducing satellite imagery complemented with a geographical information system database.

  18. Similaridade genética entre clones de seringueira (Hevea brasiliensis, por meio de marcadores RAPD Genetic similarity among rubber tree (Hevea brasiliensis clones using RAPD markers

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    Karine Cristina Bicalho

    2008-10-01

    Full Text Available A seringueira [Hevea brasiliensis (Willd. ex. Adr. de Juss Muell.-Arg.] é uma espécie nativa da região amazônica e compreende a maior fonte produtora de borracha natural do mundo. Na busca de condições mais favoráveis ao cultivo, além da busca pela auto-suficiência na produção de borracha natural, o cultivo da seringueira migrou para outras regiões do país. Objetivou-se, com o presente trabalho, estimar a similaridade genética de genótipos de seringueira, provenientes de regiões distintas do país, Lavras-MG (UFLA e Campinas-SP (IAC, por meio de marcadores moleculares RAPD. A análise foi efetuada em 41 indivíduos, representados por 17 genótipos diferentes, com base em 19 primers, que geraram 121 fragmentos polimórficos. Os dados foram analisados utilizando o software NTSYS-pc - 2.1, por meio do coeficiente de Dice e pelo método das médias (UPGMA. A similaridade genética entre o material analisado variou de 0,56 a 1,00. Na análise do dendrograma, foram observados 18 grupos. Os clones (RRIM600, GT1, PB235, PL PIM e FX2261, utilizados em diferentes repetições, foram idênticos, quando comparados entre si, entretanto o mesmo não foi observado para os clones identificados como RRIM 701. Os resultados obtidos sugerem que o material avaliado na UFLA é o mesmo implantado no IAC, exceto o RRIM 701, mostrando uma ampla variabilidade genética, disponível para estudos e propagação da cultura.The rubber tree [Hevea brasiliensis (Willd. ex. Adr. de Juss Muell.-Arg.] is a native species from Amazon region, and represents the biggest source of natural rubber in the world.. However, the rubber tree culture has had an expansion to other brazilian regions, in search of more favorable conditions for its cultivation and self-sufficiency in natural rubber. The aim of this work was to estimate genetic similarity among rubber tree clones, from different Brazilian regions, Lavras (UFLA and Campinas (IAC, by using RAPD molecular markers

  19. High levels of plasma IL-10 and expression of IL-10 by keratinocytes during visceral leishmaniasis predict subsequent development of post-kala-azar dermal leishmaniasis

    DEFF Research Database (Denmark)

    Gasim, S; Elhassan, A M; Khalil, E A

    1998-01-01

    Some patients develop post-kala-azar dermal leishmaniasis (PKDL) after they have been treated for the systemic infection kala-azar (visceral leishmaniasis). It has been an enigma why the parasites cause skin symptoms after the patients have been successfully treated for the systemic disease. We...... report here that PKDL development can be predicted before treatment of visceral leishmaniasis, and that IL-10 is involved in the pathogenesis. Before treatment of visceral leishmaniasis, Leishmania parasites were present in skin which appeared normal on all patients. However, IL-10 was detected...

  20. Genetic variability among 18 cultivars of cooking bananas and plantains by RAPD and ISSR markers

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    YUYU SURYASARI POERBA

    2010-07-01

    Full Text Available Poerba YS, Ahmad F (2010 Genetic variability among 18 cultivars of cooking bananas and plantains by RAPD and ISSR markers. Biodiversitas 11: 118-123. This study was done to assess the molecular diversity of 36 accessions (18 cultivars of the plantain and cooking bananas (Musa acuminata x M. balbisiana, AAB, ABB subgroups based on Random amplified polymorphic DNA (RAPD and and Inter Simple Sequence Repeats (ISSR markers and to determine genetic relationships in the bananas. RAPD and ISSR fingerprinting of these banana varieties was carried out by five primers of RAPDs and two primers of ISSRs. RAPD primers produced 63 amplified fragments varying from 250 to 2500 bp in size. 96.82% of the amplification bands were polymorphic. ISSR primers produced 26 amplified fragments varying from 350 bp to 2000 bp in size. The results showed that 92.86% of the amplification bands were polymorphic. The range of genetic distance of 18 cultivars was from 0.06-0.67.

  1. Molecular evaluations of thirty one clones of poplar based on RAPD and SSR molecular markers

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    Singh M.K.

    2014-01-01

    Full Text Available Poplar is an important tree species valued all over the world for its wood importance. Despite limited knowledge of the levels of genetic diversity and relatedness, their cultivation as a source of plywood is widespread. In order to facilitate reasoned scientific decisions on its management and conservation and prepare for selective breeding programme, genetic analysis of 31 genotypes was performed using RAPD and SSR molecular markers. Twenty six RAPD primers and 14 SSR primers amplified a total of 236 and 85 scoreable bands of which 86.44% and 86.02% were polymorphic. The mean coefficient of gene differentiation (Gst was 0.388 and 0.341 indicating that 61.2% and 65.9% of the genetic variation resided within the populations. Analysis of molecular variance (AMOVA indicated that majority of genetic variation (94.6% using RAPD and 89% using SSR occurred among genotypes, while the variation between the three groups (categorized as tall, medium and small plants height was 5.4% (using RAPD and 11% (using SSR. The dendrogram obtained from NJ and STRUCTURE analysis revealed splitting of genotypes into four clusters with clear distinction between short, medium and tall height genotypes, indicated that genetic differentiations measure with respect to RAPD and SSR. However, both the markers were equally useful in providing some understanding about the genetic relationship of different genotypes of poplar that are important in the conservation and exploitation of poplar genetic resources.

  2. Genetic Diversity Study Among Six Genera of Amaranth Family Found in Malang Based on RAPD Marker

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    Arik Arubil Fatinah

    2012-12-01

    Full Text Available Genera of amaranth family tend to have phenotypic variation partly caused by environmental factor. Phenotypic variation was the result of interaction between genetic and environmental factors. One of molecular markers that is widely used for detecting genetic variation is RAPD. RAPD is used for polymorphism detections and is now possible for identifiying a large number of loci and ascribes unambiguous taxonomic and genetic relationships among different taxa. Members of amaranth family found in Indonesia are Amaranthus, Celosia, Aerva, Alternanthera, Achyranthes, Gomphrena, Salsola, and Iresine. Six genera of which (Amaranthus, Celosia, Aerva, Alternanthera, Achyranthes, and Gomphrena were observed in this study. DNA was extracted from fresh young leaves using Doyle and Doyle’s method with modification in the extraction buffer used. RAPD analyses were carried out with 20 decamer primers from Kit A of Operon Technology. DNA was amplified using master cycler gradient Eppendorf with 35 cycles. RAPD products were separated on 1,5 % agarose gels and detected by staining with ethidium bromide. There were 374 bands generated in 18 random primers. The number of monomorphic bands, polymorphic bands, and the percentage of polymorphism were 21 bands, 353 bands, and 94,38 % respectively. The high number and percentage of polymorphic bands revealed genomic DNA variation. This variation is in accordance with phenotypic variation detected in this experiment. Therefore, it can be concluded that, based on DNA polymorphism detected by RAPD, Amaranth family can be classified into two sub families namely Amaranthoideae and Gomphrenoideae.

  3. Genetic diversity of Pleurotus pulmonarius revealed by RAPD, ISSR, and SRAP fingerprinting.

    Science.gov (United States)

    Yin, Yonggang; Liu, Yu; Li, Huamin; Zhao, Shuang; Wang, Shouxian; Liu, Ying; Wu, Di; Xu, Feng

    2014-03-01

    Pleurotus pulmonarius is one of the most widely cultivated and popular edible fungi in the genus Pleurotus. Three molecular markers were used to analyze the genetic diversity of 15 Chinese P. pulmonarius cultivars. In total, 21 random amplified polymorphic DNA (RAPD), 20 inter-simple sequence repeat (ISSR), and 20 sequence-related amplified polymorphism (SRAP) primers or primer pairs were selected for generating data based on their clear banding profiles produced. With the use of these RAPD, ISSR, and SRAP primers or primer pairs, a total of 361 RAPD, 283 ISSR, and 131 SRAP fragments were detected, of which 287 (79.5 %) RAPD, 211 (74.6 %) ISSR, and 98 (74.8 %) SRAP fragments were polymorphic. Unweighted Pair-Group Method with Arithmetic Mean (UPGMA) trees of these three methods were structured similarly, grouping the 15 tested strains into four clades. Subsequently, visual DNA fingerprinting and cluster analysis were performed to evaluate the resolving power of the combined RAPD, ISSR, and SRAP markers in the differentiation among these strains. The results of this study demonstrated that each method above could efficiently differentiate P. pulmonarius cultivars and could thus be considered an efficient tool for surveying genetic diversity of P. pulmonarius.

  4. Predicting death from kala-azar: construction, development, and validation of a score set and accompanying software

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    Dorcas Lamounier Costa

    Full Text Available Abstract INTRODUCTION Early identification of patients at higher risk of progressing to severe disease and death is crucial for implementing therapeutic and preventive measures; this could reduce the morbidity and mortality from kala-azar. We describe a score set composed of four scales in addition to software for quick assessment of the probability of death from kala-azar at the point of care. METHODS: Data from 883 patients diagnosed between September 2005 and August 2008 were used to derive the score set, and data from 1,031 patients diagnosed between September 2008 and November 2013 were used to validate the models. Stepwise logistic regression analyses were used to derive the optimal multivariate prediction models. Model performance was assessed by its discriminatory accuracy. A computational specialist system (Kala-Cal(r was developed to speed up the calculation of the probability of death based on clinical scores. RESULTS: The clinical prediction score showed high discrimination (area under the curve [AUC] 0.90 for distinguishing death from survival for children ≤2 years old. Performance improved after adding laboratory variables (AUC 0.93. The clinical score showed equivalent discrimination (AUC 0.89 for older children and adults, which also improved after including laboratory data (AUC 0.92. The score set also showed a high, although lower, discrimination when applied to the validation cohort. CONCLUSIONS: This score set and Kala-Cal(r software may help identify individuals with the greatest probability of death. The associated software may speed up the calculation of the probability of death based on clinical scores and assist physicians in decision-making.

  5. Molecular genetic differentiation of avian Escherichia coli by RAPD-PCR Diferenciação molecular de Escherichia coli aviária por RAPD-PCR

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    Taghi Zahraei Salehi

    2008-09-01

    Full Text Available Escherichia coli is one of the most important bacterial avian pathogens and a common inhabitant of the gastrointestinal tract of animals. Most pathogenic E. coli can not be differentiated biochemically or by classic microbiologic methods. Molecular typing methods, particularly PCR, facilitated epidemiological and ecological studies of bacteria. Here we describe the application of a random amplified polymorphic DNA- polymerase chain reaction (RAPD-PCR for molecular genetic differentiation of E. coli isolates in Iran. In this study 58 E. coli isolates including 4 standard strains, 3 food originated isolates, 33 avian isolates, 8 isolates form diarrheic calves and 10 isolates from unweaned diarrheic lambs were analyzed by RAPD-PCR using primer 1247(5/-AAG AGC CCG T-3/. The RAPD analysis showed that these isolates could be grouped into 33 RAPD types and avian isolates were discriminated into 29 genotypes. It was shown that the primer could not differentiate E. coli isolated from lambs. Discriminatory index for entire isolates was 0.912 and for avian isolates was 0.990. We concluded that RAPD-PCR can be used as a method for molecular differentiation of E. coli isolates.Escherichia coli é um dos patógenos aviários mais importantes e um habitante comum do trato gastrointestinal de animais. A maioria das cepas patogênicas não pode ser diferenciada por métodos bioquímicos ou outros métodos microbiológicos clássicos. Métodos de tipagem molecular, particularmente PCR, têm facilitado os estudos epidemiológicos e ecológicos a respeito desse microrganismo. Nesse estudo, descrevemos a aplicação do RAPD-PCR para a diferenciação molecular de isolados de E.coli do Irã. No estudo, 58 isolados, incluindo 4 isolados padrão, 3 isolados de alimentos, 33 isolados de aves, 8 isolados de bezerros diarréicos e 10 isolados de carneiros diarréicos foram analisados por RAPD-PCR com o primer 1247 (5'-AAG AGC CCG T-3'. A análise mostrou que esses

  6. Development of an RAPD-based SCAR marker for smut disease resistance in commercial sugarcane cultivars of Pakistan

    Science.gov (United States)

    Development of RAPD-derived Sequence Characterized Amplified Region (SCAR) marker in order to select Sporisorium scitamineum resistant and susceptible commercial cultivars of sugarcane from Pakistan was achieved. Bulked segregant and RAPD-analysis were conducted using 480 random decamers in initial ...

  7. Selection of Unique Escherichia coli Clones by Random Amplified Polymorphic DNA (RAPD): Evaluation by Whole Genome Sequencing

    Science.gov (United States)

    Nielsen, Karen L.; Godfrey, Paul A.; Stegger, Marc; Andersen, Paal S.; Feldgarden, Michael; Frimodt-Møller, Niels

    2014-01-01

    Identifying and characterizing clonal diversity is important when analysing fecal flora. We evaluated random amplified polymorphic DNA (RAPD) PCR, applied for selection of Escherichia coli isolates, by whole genome sequencing. RAPD was fast, and reproducible as screening method for selection of distinct E. coli clones in fecal swabs. PMID:24912108

  8. The Impact of Micro-Teaching on the Teaching Practice Performance of Undergraduate Agricultural Education Students in College of Education, Azare

    Science.gov (United States)

    Sa'ad, Tata Umar; Sabo, Shehu; Abdullahi, Aliyu Dahuwa

    2015-01-01

    Micro-teaching and teaching practices are two integral parts of teacher education programme. Therefore, this study investigated the impact of micro-teaching on the teaching practice of the undergraduate Agricultural Education Students admitted in 2012/2013 Academic session in College of Education, Azare, Bauchi State, Nigeria. The 400 level…

  9. The pathogenesis of post kala-azar dermal leishmaniasis from the field to the molecule: does ultraviolet light (UVB) radiation play a role?

    DEFF Research Database (Denmark)

    Ismail, A; Khalil, E A G; Musa, A M

    2006-01-01

    Post kala-azar dermal leishmaniasis (PKDL) is a dermatosis caused by persistence of Leishmania donovani parasites in the skin following apparently successful treatment of visceral leishmaniasis. The distribution of PKDL lesions in Sudanese patients often mirrors the clothing habits of those...

  10. Pathology of post-kala-azar dermal leishmaniasis: a light microscopical, immunohistochemical, and ultrastructural study of skin lesions and draining lymph nodes

    DEFF Research Database (Denmark)

    Ismail, Ahmed; Gadir, A Fattah A; Theander, Thor G

    2006-01-01

    BACKGROUND: Whereas the clinical manifestations and treatment of post-kala-azar dermal leishmaniasis (PKDL) have been adequately described before, the pathology received little attention, particularly the African form of PKDL which shows some clinical differences from the disease in India...... leishmaniasis: a light microscopical, immunohistochemical, and ultrastructural study of skin lesions and draining lymph nodes....

  11. Detection and characterization of Leishmania in tissues of patients with post kala-azar dermal leishmaniasis using a specific monoclonal antibody

    DEFF Research Database (Denmark)

    Ismail, A; Kharazmi, A; Permin, H

    1997-01-01

    Sections from skin lesions and draining lymph nodes of patients with post kala-azar dermal leishmaniasis were examined using an immunoperoxidase method and a monoclonal antibody directed against Leishmania donovani. Parasites were detected in 22 of 25 biopsies (88%). In parallel sections stained...

  12. Molecular characterization of eight Indian Snakehead species (Pisces: Perciformes Channidae) using RAPD markers.

    Science.gov (United States)

    Bhat, Ajaz Ali; Haniffa, M A; Divya, P R; Gopalakrishnan, A; Milton, M James; Kumar, Raj; Paray, Bilal Ahmad

    2012-04-01

    Murrels (Perciformes; Channidei; Channidae) are unique group of freshwater air breathing fishes having a confined distribution to African and Asian continents. The phylogenetic relationship among eight Channid species viz. Channa aurantimaculata, Channa bleheri, Channa diplogramma, Channa gachua, Channa marulius, Channa punctatus, Channa stewartii and Channa striatus were investigated using RAPD markers. Eight random oligodecamers viz. OPAC03, OPAC05, OPAC07, OPAC09, OPAC19, OPA10, OPA11 and OPA16 were used to generate the RAPD profile. Estimates of Nei's (Genetics, 89:583-590, 1978) unbiased genetic distance (D) demonstrated sufficient genetic divergence to discriminate the samples of different species and the values ranged from 0.3292 to 0.800 The present RAPD analyses strongly substantiate the view of earlier morphological and osteological studies of Channid species, the closer association among species in "gachua" and "marulius" groups.

  13. STUDY ON GERMPLASMIC RESOURCES OF LYCORIS LONGITUBA USING RAPD AND ISSR

    Directory of Open Access Journals (Sweden)

    Deng CHUANLIANG

    2006-08-01

    Full Text Available The perianth DNA extraction methods were discussed as Lycoris longituba for example. By means of RAPD and ISSR, germplasmic resources of Lycoris longituba were primarily studied. The results were as follow: by RAPD, a total of 77 discernible loci were obtained using 12 primers, of which 53 loci were polymorphic (PPB = 68.8%; by ISSR, 67 discernible loci were got using 9 primers, of which 62 loci were polymorphic (PPB = 92.5%. So, genetic diversity of Lycoris longituba was abundant, whose germplasmic resources could be stored for breeding. From UPGMA dendrogram of Lycoris longituba using RAPD or ISSR method, three Lycoris longituba types were supported with molecular evidence, which were originally distinguished by flower color. Therefore, in the future use of Lycoris longituba germplasmic resources, different varieties of Lycoris longituba could be cultivated.

  14. IDENTIFICATION OF RAPD MARKER LINKED TO POWDERY MILDEW RESISTANCE GENE Pm 12 IN WHEAT

    Institute of Scientific and Technical Information of China (English)

    GU Feng; ZHANG Qing-li; GUO Xiao-chun; LI Tao; WANG Hong-gang

    2004-01-01

    Powdery mildew is one of the most serious diseases of wheat in China. In this paper,bulked segregant analysis (BSA) was used to search for randomly amplified polymorphic DNA (RAPD) markers linked to the Pm12 gene,which confers resistance to the powdery mildew in wheat. 200 decamer primers were screened and one RAPD marker (S1071900) was identified to be linked to Pm 12 in coupling phase, and their genetic distance is 11.98 ±4.00cM. This marker can be used for marker - assisted selection in wheat breeding for the identification or pyramiding of Pm12 with other resistance genes.Key Words: Wheat, RAPD, Powdery mildew, resistance genes.

  15. Assessing phylogenetic relationships of Lycium samples using RAPD and entropy theory

    Institute of Scientific and Technical Information of China (English)

    Xiao-lin YIN; Kai-tai FANG; Yi-zeng LIANG; Ricky NS WONG; Amber WY HA

    2005-01-01

    Aim: To evaluate the phylogenetic relationships among related species of Lycium samples. Methods: Random amplified polymorphic DNA (RAPD) fingerprinting and lab-on-a-chip electrophoresis techniques were used to analyze the characteristics of Lycium species. Seven species and 3 varieties of Lycium were studied.Based on RAPD fingerprint data obtained from 11 primers, we proposed a new index, called dispersivity, using entropy theory and projection methods to depict the diversity of the DNA fingerprints. Results: Using the proposed dispersivity,primers were sorted and the dendrograms of the 7 species and 3 varieties of Lycium were constructed synthetically by merging primer information. Conclusion:Phylogenetic relationships among Lycium samples were constructed synthetically based on RAPD fingerprint data generated from 11 primers.

  16. Molecular typing of Epidermophyton floccosum isolated from patients with dermatophytosis by RAPD-PCR.

    Science.gov (United States)

    Khosravi, Alireza; Behzad, Forough; Sabokbar, Azar; Shokri, Hojjatollah; Haddadi, Siamak; Masoudi-Nejad, Ali

    2010-12-01

    We evaluated the ability of randomly amplified polymorphic DNA (RAPD) to type Epidermophyton floccosum isolates recovered from patients with dermatophytosis originating from different regions of Iran. A total of 13 clinical isolates of E. floccosum obtained from Iranian patients were analyzed by RAPD with 7 arbitrary primers (OPN16, OPD18' OPU15, OPX19, R28, OPA04 and OPAA17). Among the applied primers, OPN16 produced banding patterns from all the isolates. In addition, some of the isolates had very close relation. The phenon line which represented the mean similarities was at the value of 73%. At this level, 4 groups were characterized. Two isolates of a patient had different molecular patterns, suggesting infection transmission from different sources in the case of a single patient. RAPD-PCR provided a rapid and practical tool for identification of E. floccosum isolates, which was independent of morphological characteristics, and enhanced laboratory diagnosis of dermatophytosis.

  17. Genome relationship among nine species of Millettieae (Leguminosae: Papilionoideae) based on random amplified polymorphic DNA (RAPD).

    Science.gov (United States)

    Acharya, Laxmikanta; Mukherjee, Arup Kumar; Panda, Pratap Chandra

    2004-01-01

    Random amplified polymorphic DNA (RAPD) marker was used to establish intergeneric classification and phylogeny of the tribe Millettieae sensu Geesink (1984) (Leguminosae: Papilionoideae) and to assess genetic relationship between 9 constituent species belonging to 5 traditionally recognized genera under the tribe. DNA from pooled leaf samples was isolated and RAPD analysis performed using 25 decamer primers. The genetic similarities were derived from the dendrogram constructed by the pooled RAPD data using a similarity index, which supported clear grouping of species under their respective genera, inter- and intra-generic classification and phylogeny and also merger of Pongamia with Millettia. Elevation of Tephrosia purpurea var. pumila to the rank of a species (T. pumila) based on morphological characteristics is also supported through this study of molecular markers.

  18. Use of RAPD and PCR double amplification in the study of ancient DNA

    Directory of Open Access Journals (Sweden)

    F. Balzano

    2011-01-01

    Full Text Available This project analysed the DNA extracted from bones of ancient sheep which have been brought to light in Sardinian different archaeological sites. In order to better analyse this highly fragmented DNA, a double amplification technique was chosen. The first approach consisted of RAPD-PCR abd the second one in classic PCR. The RAPD-PCR amplified random fragments and allowed the production of numerous amplicons. The products of RAPD amplification have been amplified, more specifically, by the second PCR using primers for a sequence of 176 bp of mitochondrial D-loop region. These DNA fragments have been sequenced and the sequence analysis has confirmed that it belonged to Ovis aries. Consequently, this provedure can be considered a valid tool to perform amplification of degraded DNA, such as ancient DNA.

  19. 甘草RAPD-PCR反应体系正交优化研究%Optimization of RAPD-PCR Reaction System for Glycyrrhiza uralensis Based on Orthogonal Design

    Institute of Scientific and Technical Information of China (English)

    张增福; 董建力; 李明

    2011-01-01

    [Objective] The aim was to obtain the optimum RAPD-PCR reaction system for Glycyrrhiza uralensis. [ Method] Orthogonal design was adopted to screen the suitable concentration of major factors (dNTPs, primer, Taq polymerase, DNA template) in PCR reaction system. [ Result] The optimal protocol was accomplished by orthogonal design in 25 μl reaction volume containing 10 × PCR buffer solution(include MgCl2) 2.5 μl, 10 mmol/L dNTPs 2.5 μl, 50 ng DNA template 2.0 μl, 10 μmol/L primer 2. 0 μl, 5 U/μl Taq polymerase 0.4 μl; the optimum annealing temperature was 34 ℃. [ Conclusion] Orthogonal design was an effective method for the optimization of RAPD-PCR reaction system for C. Uralensis.%[目的]建立一套适合甘草分子学研究的RAPD-PCR反应体系.[方法]以甘草种质为试材,采用正交试验法设计,对影响RAPD-PCR扩增的主要因素dNTPs、引物、Taq酶和DNA模板进行优化筛选.[结果]总体积25μl的甘草RAPD-PCR最佳反应体系为:10 ×PCR缓冲液(含MgCl2 )2.5 μl,10 mmol/L dNTPs 2.5 μl,50 ng DNA 2.0μl,10 μmol/L引物2.0μl,5 U/μl Taq酶0.4μl.对引物的退火温度进行了梯度筛选,34℃时扩增效果较好.[结论]进行甘草RAPD-PCR反应体系的正交优化非常有效.

  20. Genetic similarity among strawberry cultivars assessed by RAPD and ISSR markers

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    Rafael Gustavo Ferreira Morales

    2011-12-01

    Full Text Available Most strawberry (Fragaria × ananassa Duchesne cultivars used in Brazil are developed in other countries, it became clear the need to start the strawberry breeding program in the country. To start a breeding program is necessary the genetic characterization of the germplasm available. Molecular markers are important tools that can be used for this purpose. The objectives of the present study were to assess the genetic similarity among 11 strawberry cultivars using RAPD and ISSR molecular markers and to indicate the possible promising crosses. The DNA of the eleven strawberry cultivars was extracted and amplified by PCR with RAPD and ISSR primers. The DNA fragments were separated in agarose gel for the RAPD markers and in polyacrylamide gel for the ISSR markers. The genetic similarity matrix was estimated by the Jaccard coefficient. Based on this matrix, the cultivars were grouped using the UPGMA method. The dendogram generated by the RAPD markers distributed the cultivars in three groups while the ISSR markers generated two groups. There was no direct relationship between the marker groups when the two types of markers were compared. The grouping proposed by the ISSR markers was more coherent with the origin and the genealogy of the cultivars than that proposed by the RAPD markers, and it can be considered the most efficient method for the study of genetic divergence in strawberry. The most promising crosses, based on the genetic divergence estimated from the RAPD and ISSR molecular data were between the Tudla and Ventana and the Oso Grande and Ventana cultivars, respectively.

  1. Variation in Salmonella enteritidis RAPD-PCR patterns may not be due to genetic differences.

    Science.gov (United States)

    Mathis, Demetrius L; Berghaus, Roy D; Lee, Margie D; Maurer, John J

    2011-12-01

    Salmonella Enteritidis is a leading cause of gastroenteritis associated with consumption of contaminated poultry meat and eggs. Because pulsed-field gel electrophoresis (PFGE) has limited utility in distinguishing between clonal Salmonella Enteritidis isolates, random amplified polymorphic DNA (RAPD) PCR has been recommended as an alternative molecular fingerprinting tool. This study's objective was to determine whether increasing PCR stringency would improve the repeatability of RAPD DNA patterns based on assessment of target sites within the genome. An in silico PCR was performed to predict amplification products from an Salmonella Enteritidis genome sequence for three different RAPD primers (1247, 1283, and OPA4) and to determine whether any primer would be more likely to amplify variable regions within the genome. A comparison of within- and between-isolate similarities in RAPD patterns was performed using primer 1247, which was predicted by in silico analysis to yield a variable size range of amplicons. In order to reduce artifactual variability associated with the method, three different methods for template preparation were evaluated. All were found to provide comparable results with respect to the similarities observed with repeated analyses of the same Salmonella Enteritidis isolates (n = 18, P = 0.91). Although the median within-isolate similarity (76.0%) was significantly greater than the median between-isolate similarity (66.7%; P = 0.001), duplicate RAPD-PCR runs of the same Salmonella Enteritidis isolates produced DNA patterns that ranged in similarity between 61.5 and 100%. These results indicate that the repeatability of RAPD-PCR is insufficient to distinguish genetic differences among related and unrelated Salmonella Enteritidis isolates.

  2. Genes that encodes NAGT, MIF1 and MIF2 are not virulence factors for kala-azar caused by Leishmania infantum

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    Bruno Guedes Alcoforado Aguiar

    2014-10-01

    Full Text Available Introduction Kala-azar is a disease resulting from infection by Leishmania donovani and Leishmania infantum. Most patients with the disease exhibit prolonged fever, wasting, anemia and hepatosplenomegaly without complications. However, some patients develop severe disease with hemorrhagic manifestations, bacterial infections, jaundice, and edema dyspnea, among other symptoms, followed by death. Among the parasite molecules that might influence the disease severity are the macrophage migration inhibitory factor-like proteins (MIF1 and MIF2 and N-acetylglucosamine-1-phosphotransferase (NAGT, which act in the first step of protein N-glycosylation. This study aimed to determine whether MIF1, MIF2 and NAGT are virulence factors for severe kala-azar. Methods To determine the parasite genotype in kala-azar patients from Northeastern Brazil, we sequenced the NAGT genes of L. infantum from 68 patients as well as the MIF1 and MIF2 genes from 76 different subjects with diverse clinical manifestations. After polymerase chain reaction (PCR, the fragments were sequenced, followed by polymorphism identification. Results The nucleotide sequencing of the 144 amplicons revealed the absence of genetic variability of the NAGT, MIF1 and MIF2 genes between the isolates. The conservation of these genes suggests that the clinical variability of kala-azar does not depend upon these genes. Additionally, this conservation suggests that these genes may be critical for parasite survival. Conclusions NAGT, MIF1 and MIF2 do not alter the severity of kala-azar. NAGT, MIF1 and MIF2 are highly conserved among different isolates of identical species and exhibit potential for use in phylogenetic inferences or molecular diagnosis.

  3. Assessment of genetic diversity among different indigenous Xanthomonas isolates via RAPD and ISSR

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    Fatima Sabin

    2012-01-01

    Full Text Available The genetic diversity among seven Xanthomonas isolates representing four species was assessed using RAPD and ISSR PCR-based techniques. Both techniques revealed high degrees of polymorphisms among the studied isolates. A cluster dendrogram based on the combined data of RAPD and ISSR showed that genetic diversity exists in local isolates of Xanthomonas. In terms of percentage similarity values, the genomic variation was found to be in the range of 29.29% - 100% among the isolates. X. campestris (Mangifera indica remained unclustered in cluster dendrogram and revealed a unique genomic profile compared to other isolates used in this study.

  4. Molecular characterization of Desmodium species--an important ingredient of 'Dashmoola' by RAPD analysis.

    Science.gov (United States)

    Irshad, Saba; Singh, Jyotsna; Kakkar, Poonam; Mehrotra, Shanta

    2009-03-01

    Identification of medicinal plants by their molecular signature is a fast growing tool. The identification of Desmodium gangeticum (L.) DC. (Shalparni, a constituent of Ayurvedic formulation "Dashmoolarishtha") was carried out using genomic approach. Authentic samples of D. gangeticum(L.) DC., D. velutinum (Willd.) DC. and D. triflorum (L.) DC. were analyzed and compared to commercial samples of various origin. Within twenty primers used, eleven gave 223 RAPD fragments. RAPD profiles of three species showed very low similarity index (0.21-0.39), whereas market samples showed high similarity of 0.82-0.89 with authenticated D. gangeticum.

  5. Identification of Specific RAPD Markers Linked to Anthracnose Resistant Gene in Native Wild Grapes of China

    Institute of Scientific and Technical Information of China (English)

    WANG Xi-ping; WANG Yue-jin; ZHOU Peng; ZHENG Xue-qin

    2001-01-01

    Randomly amplified polymorphic DNA (RAPD) was employed to detect molecular markers linked to anthracnose ( Spheceloma ampelinum de Bary) resistant gene in the native wild grapes ( Vitis L. ) of China. RAPD marker OPJ13-300 was linked to anthracnose resistant gene using 90-3 cross F1 V. quinquangularis Rehd (shang-24) × V. vinifera (Longyan). The marker was verified in 90-3 cross F1, Chinese wild grapes and V. riparia and European grape cuitivars. This work has provided a solid basis for molecular marker-assisted selection (MAS) to disease resistance and cloning of disease resistant genes.

  6. An Effective Procedure for the RAPD Analysis of Hemp Cannabissativa L.

    Institute of Scientific and Technical Information of China (English)

    SongShujuan; Robert.C.ClarkeI; ShaoHong

    2001-01-01

    China has a great resource of Cannabis. Research on the taxonomy and morphology of Chinese Cannabis has been carried out, but so far no molecular genetic research has been published. Random amplifiedpolymorphic DNA (RAPD) is a suitable technique for molecular genetic research on Cannabis. In tills experiment,using Cannabis herbarium specimens as a source of genetic materials, the correlative conditions of the Polymerasechain reaction (PCR), (i.e., gradient density of Mg2+, dNTPs, Taq DNA polymerase, anneal temperature, anneal timeand reaction cycles) were examined separately. An effective procedure for the RAPD analysis of Cannabis wasobtained.

  7. Use of RAPD molecular markers on differentiation of brazilian and chinese Ganoderma lucidum strains

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    Leonardo do Nascimento Rolim

    2011-04-01

    Full Text Available The aim of this work was to analyze the Brazilian and Chinese strains of Ganoderma lucidum with molecular RAPD markers. A similarity matrix was elaborated and the RAPD profiles of G. lucidum strains were also compared to two other Ganoderma spp: G. applanatum and G. lipsiense in order to produce genetic similarity among the species. Based on the primers used, it was possible to determine that the Brazilian strains and Chinese strain CC-22 are alike. The method and the primers selection showed to be appropriate for the genetic identification of G. lucidum strains, enabling them to be improved and used in research, as well as in the world market.

  8. Analysis of genetic diversity of certain species of Piper using RAPD-based molecular markers.

    Science.gov (United States)

    Chowdhury, Utpal; Tanti, Bhaben; Rethy, Parakkal; Gajurel, Padma Raj

    2014-09-01

    The utility of RAPD markers in assessing genetic diversity and phenetic relationships of six different species of Piper from Northeast India was investigated. Polymerase chain reaction (PCR) with four arbitrary 10-mer oligonucleotide primers applied to the six species produced a total of 195 marker bands, of which, 159 were polymorphic. On average, six RAPD fragments were amplified per reaction. In the UPGMA phenetic dendrogram based on Jaccard's coefficient, the different accessions of Piper showed a high level of genetic variation. This study may be useful in identifying diverse genetic stocks of Piper, which may then be conserved on a priority basis.

  9. Fronteras del corazón shamánico: Azares y dilemas Qom

    OpenAIRE

    2010-01-01

    En este artículo analizo dilemas y paradojas que plantea la modernidad en el caso de los Takshek Qom o tobas del oriente de Formosa (Argentina). Para ello me centro en la experiencia del lkillakte o corazón como uno de los núcleos vitales y existenciales de la persona. Allí se alojan las distintas clases de poder shamánico y también es sede de los pensamientos y las emociones. Me interesa explorar qué le pasa en términos simbólicos al corazón shamánico frente al contacto con el catolicismo y ...

  10. Optimización y control de juegos de azar y motores brownianos colectivos

    OpenAIRE

    Dinis Vizcaíno, Luis Ignacio

    2011-01-01

    En esta tesis se han estudiado aspectos fundamentales relacionados con la rectificación de fluctuaciones térmicas: el control y optimización de motores colectivos, formados por un gran número de partículas brownianas. Hemos analizado diversos protocolos de control que se basan en la información sobre la posición de las partículas para mejorar el transporte. Al menos uno de estos protocolos podría utilizarse en una realización experimental de un motor browniano en el que las partículas son mon...

  11. Optimización y control de juegos de azar y motores brownianos colectivos

    OpenAIRE

    Dinis Vizcaíno, Luis Ignacio

    2005-01-01

    En esta tesis se han estudiado aspectos fundamentales relacionados con la rectificación de fluctuaciones térmicas: el control y optimización de motores colectivos, formados por un gran número de partículas brownianas. Hemos analizado diversos protocolos de control que se basan en la información sobre la posición de las partículas para mejorar el transporte. Al menos uno de estos protocolos podría utilizarse en una realización experimental de un motor browniano en el que las partículas son mon...

  12. Application of random amplified polymorphic DNA (RAPD) analysis coupled with microchip electrophoresis for high-resolution identification of Monascus strains.

    Science.gov (United States)

    Shinzato, Naoya; Namihira, Tomoyuki; Tamaki, Yasutomo; Tsukahara, Masatoshi; Matsui, Toru

    2009-04-01

    Monascus fungi are commonly used for a variety of food products in Asia, and are also known to produce some biologically active compounds. Since the use of Monascus is expected to increase in food industries, strain-level identification and management of Monascus will be needed in the near future. In the present study, random amplified polymorphic DNA (RAPD) analysis coupled with microchip electrophoresis was applied for this purpose. Evaluations of the analysis stability revealed that reproducible results could be obtained, although template DNA fragmentation could influence the resulting RAPD pattern. RAPD analysis using 15 Monascus strains consisting of four species, M. ruber, M. pilosus, M. purpureus, and M. kaoliang showed that each strain generated a unique RAPD pattern, which allows strain-level identification of Monascus. In addition, the phylogenetic tree constructed from RAPD patterns reflected M. ruber-M. pilosus and M. purpureus-M. kaoliang clusters inferred from both ITS and beta-tubulin gene sequences, which indicated that the RAPD pattern could reflect their phylogenetic traits to a certain extent. On the other hand, RAPD analysis did not support the monophyletic clustering of the four Monascus species used in this study, which suggests the necessity of reexamination of species boundaries in Monascus.

  13. Identification of RAPD markers and their use for molecular mapping in pea (Pisum sativum L.).

    Science.gov (United States)

    Cheghamirza, Kianoosh; Koveza, Oksana; Konovalov, Fedor; Gostimsky, Sergei

    2002-01-01

    The RAPD method (Random Amplified Polymorphic DNA) was used for identifying and mapping new molecular markers in pea. RAPD analysis of various cultivars and lines of pea was carried out using 10-mer random primers. The presence of multiple polymorphism between cultivars and lines was revealed; at least one fragment for any given primer was present in the DNA of one form of pea and absent in the DNA of another line or cultivar. To detect molecular markers linked to the genes of chi-15, xa-18 and also to the 12 morphological markers of the L-1238 line, the F2 populations (Chi-15 ? L-1238), (Vio ? L-1238), (Xa-18 ? L-1238), (L-111 ? Chi-15) and (L-84 ? Xa-18) were studied via bulked segregant analysis. DNA molecular analysis of F1 hybrids revealed the presence of parental polymorphic fragments in all of the populations. The study of the F2 plants showed that the obtained fragments are inherited as Mendelian factors. 13 RAPD-markers linked to genes of A/a (flower color), I/i (seed color), Gp/gp (pod color), R/r (seed form), S/s (seeds linkage), and also to genes of Chi-15/chi-15 (leaf color) and Xa-18/xa-18 (leaf color) were discovered. The study of individual plant DNA from the F2 populations allowed us to determine the genetic distances between genes and the RAPD markers linked to them.

  14. Evaluation of genetic diversity of Clinacanthus nutans (Acanthaceaea) using RAPD, ISSR and RAMP markers.

    Science.gov (United States)

    Ismail, Noor Zafirah; Arsad, Hasni; Samian, Mohammed Razip; Ab Majid, Abdul Hafiz; Hamdan, Mohammad Razak

    2016-10-01

    Three polymerase chain reaction (PCR) techniques were compared to analyse the genetic diversity of Clinacanthus nutans eight populations in the northern region of Peninsular Malaysia. The PCR techniques were random amplified polymorphic deoxyribonucleic acids (RAPD), inter-simple sequence repeats (ISSR) and random amplified microsatellite polymorphisms (RAMP). Leaf genomic DNA was PCR amplified using 17 RAPD, 8 ISSR and 136 RAMP primers . However, only 10 RAPD primers, 5 ISSR primers and 37 RAMP primers produced reproducible bands. The results were evaluated for polymorphic information content (PIC), marker index (MI) and resolving power (RP). The RAMP marker was the most useful marker compared to RAPD and ISSR markers because it showed the highest average value of PIC (0.25), MI (11.36) and RP (2.86). The genetic diversity showed a high percentage of polymorphism at the species level compared to the population level. Furthermore, analysis of molecular variance revealed that the genetic diversity was higher within populations, as compared to among populations of C. nutans. From the results, the RAMP technique was recommended for the analysis of genetic diversity of C. nutans.

  15. Intraspecific variability of Bipolaris sorokiniana isolates determined by random-amplified polymorphic DNA (RAPD).

    Science.gov (United States)

    de Oliveira, Andréia M R; Matsumura, Aida T S; Prestes, Ariano M; Van Der Sand, Sueli T

    2002-01-01

    Isolates of Bipolaris sorokiniana were analyzed by random-amplified polymorphic DNA (RAPD) techniques to determine the amount of intraspecific genetic variability and to study host-pathogen interactions. Ten isolates originated from different regions of Brazil were examined. Plants of the wheat cultivars BR8, BH1146 (original host) and IAC-5 Maringá, classified as resistant, moderately resistant or susceptible to B. sorokiniana, respectively, were inoculated with these 10 isolates. Twenty-seven isolates were recovered from these cultivars and were analyzed by RAPD assay and compared to the RAPD of the original 10 isolates. According to the RAPD profiles there was a high level of genetic variability among the isolates. We detected 69 polymorphic fragments, ranging from 1.6 to 0.54 kb, in the original 10 isolates; 57 fragments with sizes between 1.98 and 0.38 kb from the isolates recovered from BH1146; 47 polymorphic bands, ranging from 1.96-0.54 kb, were detected in the isolates from BR8 and 32 fragments between 1.98 and 0.42 kb in isolates were recovered from IAC-5 Maringá. The number of polymorphic fragments varied, even for the same isolate, when the isolates were recovered from different cultivar hosts.

  16. Genetic Diversity and Relationships among Diploid and Tetraploid Cotton Using RAPD Markers

    Institute of Scientific and Technical Information of China (English)

    MOGHADDAM Mohammad Reza Ramazani; MAJIDI Eslam; MOFIDABADI Ali Jafari

    2008-01-01

    @@ Gossypium species (+49) represent a vast resource of genetic diversity for improvement of cultivated cotton.To determine intra- and inter-specific genetic diversity and relationships,we employed 30 RAPD random detainer primers on twenty eight cotton accessions from 2 diploid cultivated species (G.arboreum,G.herbaceum) and 2 tetraploid cultivated species (G.hirsuturn,G.barbadense).

  17. Characterization of Specific RAPD Markers of Virulence in Trichomonas vaginalis Isolates

    Science.gov (United States)

    FRAGA, Jorge; ROJAS, Lázara; SARIEGO, Idalia; FERNÁNDEZ-CALIENES, Aymé

    2015-01-01

    Background: As for human trichomoniasis the host-parasite relationship is very complex, and the broad ranges of clinical symptoms are unlikely be attributable to a single pathogenic mechanism. Specific Random Amplified Polymorphic DNA (RAPD) markers of 490 bp, 720 bp and 460 bp using the primers Tv-5, OPA-6 and OPA-11, respectively, were reported. This was the first description of possible genetic virulence markers of the infection by T. vaginalis. The aim of this study was to characterize the specific RAPD markers in order to elucidate their importance on virulence of this illness. Methods: The selected specific RAPD fragments were cloned and sequenced. The obtained sequences were compared by the BLAST algorithm. Results: The nucleotide sequence of the Tv-5490 RAPD marker exhibited significant similarity to T. vaginalis hypothetical G3 leucine rich repeat (LRR) family protein (e-value: 6e-14) and Giardia lamblia leucine rich repeat protein 1 virus receptor protein (e-value: 6e-14 and 2e-12) ; however, the OPA-6720 and OPA-11460 showed no significant similarity with any coding published sequence. All the evaluated strains showed the presence of the LRR gene. Conclusion: These results demonstrate a possible role of this gene in the virulence of T. vaginalis and in the parasite infection with Trichomonas virus as a possible virus receptor. Further analysis of this gene and encoded protein will allow determining the role that they play in the isolates virus susceptible or resistant phenotypes. PMID:26622300

  18. KERAGAMAN GENETIK AKSESI EKINASE (Echinacea purpurea (L. Moench HASIL SELEKSI MASSA TAHUN I MELALUI ANALISIS RAPD

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    Dyah Subositi

    2013-07-01

    Full Text Available Ekinase (Echinacea purpurea (L. Moench a medicinal plant that has immunostimulatory activity. This plant has been cultivating in Tawangmangu region by Medicinal Plant and Traditional Medicine Research and Development Office since 2002. Ten accessions of E. purpurea were found based on their morphological variation, three accessions of them are selected as promising accessions namely BH2, BHU3 dan BHU5. The objective of this research was to observe the genetic diversity of  those  promising accessions and 8 variants accession from mass selection year I using RAPD analysis. Those accessions were amplified using 10 RAPD primers. A total of 64 scorable fragments were generated from 9 RAPD primers, among which 48 fragments (75% were polymorphic. The Dice coefficient was used to calculated the genetic similarity and UPGMA was used to generate the dendogram. The genetic similarity index among accessions evaluated ranged from 75,49-84,21% thus indicating that low level of genetic diversity. RAPD analysis proved to be efficient for genetic diversity of ekinase accessions from mass selection year I.

  19. RAPD-SCAR marker and genetic relationship analysis of three Demodex species (Acari: Demodicidae).

    Science.gov (United States)

    Zhao, Ya-E; Wu, Li-Ping

    2012-06-01

    For a long time, classification of Demodex mites has been mainly based on their hosts and phenotype characteristics. The study was the first to conduct molecular identification and genetic relationship analysis for six isolates of three Demodex species by random amplified polymorphic DNA (RAPD) and sequence-characterized amplified region (SCAR) marker. Totally, 239 DNA fragments were amplified from six Demodex isolates with 10 random primers in RAPD, of which 165 were polymorphic. Using a single primer, at least five fragments and at most 40 in the six isolates were amplified, whereas within a single isolate, a range of 35-49 fragments were amplified. DNA fingerprints of primers CZ 1-9 revealed intra- and interspecies difference in six Demodex isolates, whereas primer CZ 10 only revealed interspecies difference. The genetic distance and dendrogram showed the intraspecific genetic distances were closer than the interspecific genetic distances. The interspecific genetic distances of Demodex folliculorum and Demodex canis (0.7931-0.8140) were shorter than that of Demodex brevis and D. canis (0.8182-0.8987). The RAPD-SCAR marker displayed primer CZ 10 could be applied to identify the three Demodex species. The 479-bp fragment was specific for D. brevis, and the 261-bp fragment was specific for D. canis. The conclusion was that the RAPD-SCAR multi-marker was effective in molecular identification of three Demodex species. The genetic relationship between D. folliculorum and D. canis was nearer than that between D. folliculorum and D. brevis.

  20. Genetic diversity and relationships in mulberry (genus Morus as revealed by RAPD and ISSR marker assays

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    Thangavelu K

    2004-01-01

    Full Text Available Abstract Background The genus Morus, known as mulberry, is a dioecious and cross-pollinating plant that is the sole food for the domesticated silkworm, Bombyx mori. Traditional methods using morphological traits for classification are largely unsuccessful in establishing the diversity and relationships among different mulberry species because of environmental influence on traits of interest. As a more robust alternative, PCR based marker assays including RAPD and ISSR were employed to study the genetic diversity and interrelationships among twelve domesticated and three wild mulberry species. Results RAPD analysis using 19 random primers generated 128 discrete markers ranging from 500–3000 bp in size. One-hundred-nineteen of these were polymorphic (92%, with an average of 6.26 markers per primer. Among these were a few putative species-specific amplification products which could be useful for germplasm classification and introgression studies. The ISSR analysis employed six anchored primers, 4 of which generated 93 polymorphic markers with an average of 23.25 markers per primer. Cluster analysis of RAPD and ISSR data using the WINBOOT package to calculate the Dice coefficient resulted into two clusters, one comprising polyploid wild species and the other with domesticated (mostly diploid species. Conclusion These results suggest that RAPD and ISSR markers are useful for mulberry genetic diversity analysis and germplasm characterization, and that putative species-specific markers may be obtained which can be converted to SCARs after further studies.

  1. Genetic diversity in natural populations of Jacaranda decurrens Cham. determined using RAPD and AFLP markers

    Directory of Open Access Journals (Sweden)

    Bianca W. Bertoni

    2010-01-01

    Full Text Available Jacaranda decurrens (Bignoniaceae is an endemic species of the Cerrado with validated antitumoral activity. The genetic diversity of six populations of J. decurrens located in the State of São Paulo was determined in this study by using molecular markers for randomly amplified polymorphic DNA (RAPD and amplified fragment length polymorphism (AFLP. Following optimization of the amplification reaction, 10 selected primers generated 78 reproducible RAPD fragments that were mostly (69.2% polymorphic. Two hundred and five reproducible AFLP fragments were generated by using four selected primer combinations; 46.3% of these fragments were polymorphic, indicating a considerable level of genetic diversity. Analysis of molecular variance (AMOVA using these two groups of markers indicated that variability was strongly structured amongst populations. The unweighted pair group method with arithmatic mean (UPGMA and Pearson's correlation coefficient (RAPD -0.16, p = 0.2082; AFLP 0.37, p = 0.1006 between genetic matrices and geographic distances suggested that the population structure followed an island model in which a single population of infinite size gave rise to the current populations of J. decurrens, independently of their spatial position. The results of this study indicate that RAPD and AFLP markers were similarly efficient in measuring the genetic variability amongst natural populations of J. decurrens. These data may be useful for developing strategies for the preservation of this medicinal species in the Cerrado.

  2. 白僵菌RAPD-PCR反应体系优化%RAPD-PCR Reaction System for Beauveria bassiana

    Institute of Scientific and Technical Information of China (English)

    魏晓鹏; 李会平; 黄大庄; 唐秀光

    2012-01-01

    为了建立一套适宜于白僵菌(Beauveria bassiana)退化研究的RAPD-PCR反应体系及反应程序,通过采用L16( 45)正交试验及退火温度和循环次数的单因素优化对反应体系中的各因素进行优化组合.结果表明:20μL PCR反应体系及反应程序中各因素优化组合为,10×Buffer 2 μL,MgCl2(25 mmol/L)2.4 μL,4种dNTP(各2.5mmol/L)0.8 μL,随机引物(10 μmol/L)1.4 μL,TaqDNA聚合酶(5 U/μL)0.4 μL,模板DNA( 10 mg/L)1 μL.反应条件为,94℃预变性2 min,94℃变性30 s,38℃退火40 s,72℃延伸1 min,循环次数40次,72℃延伸5 min.%An experiment was conducted to establish an optimal PCR (polymerase chain reaction) reaction system and procedure for the degradation of Beauveria bassiana. Single factor test and L16(45) orthogonal experiment were used to optimize the combination of factors for the reaction system. The optimum factor combination was obtained with 20 μL reaction volume containing 2 μL l0×Buffer, 2.4 μL MgCl2(25 mmol/L) , 0. 8 μL four types of dNTPs (each 2. 5 mmol/L) , 1.4μL random primer (10 μmol/L) , 0.4 μL Taq polymerase (5 U/ μL) , and 1 μL template DNA (10 mg/L). Reaction conditions were as follows; predenaturing at 94 degrees C for 2 min, followed by 40 cycles of denaturing at 94 degrees C for 30 s, annealing at 38 degrees C for 40 s, extension at 72 degrees C for 1 min, and final extension at 72 degrees C for 5 min.

  3. A genetic linkage map for hazelnut (Corylus avellana L.) based on RAPD and SSR markers.

    Science.gov (United States)

    Mehlenbacher, Shawn A; Brown, Rebecca N; Nouhra, Eduardo R; Gökirmak, Tufan; Bassil, Nahla V; Kubisiak, Thomas L

    2006-02-01

    A linkage map for European hazelnut (Corylus avellana L.) was constructed using random amplified polymorphic DNA (RAPD) and simple sequence repeat (SSR) markers and the 2-way pseudotestcross approach. A full-sib population of 144 seedlings from the cross OSU 252.146 x OSU 414.062 was used. RAPD markers in testcross configuration, segregating 1:1, were used to construct separate maps for each parent. Fifty additional RAPD loci were assigned to linkage groups as accessory markers whose exact location could not be determined. Markers in intercross configuration, segregating 3:1, were used to pair groups in one parent with their homologues in the other. Eleven groups were identified for each parent, corresponding to the haploid chromosome number of hazelnut (n = x = 11). Thirty of the 31 SSR loci were able to be assigned to a linkage group. The maternal map included 249 RAPD and 20 SSR markers and spanned a distance of 661 cM. The paternal map included 271 RAPD and 28 SSR markers and spanned a distance of 812 cM. The maps are quite dense, with an average of 2.6 cM between adjacent markers. The S-locus, which controls pollen-stigma incompatibility, was placed on chromosome 5S where 6 markers linked within a distance of 10 cM were identified. A locus for resistance to eastern filbert blight, caused by Anisogramma anomala, was placed on chromosome 6R for which two additional markers tightly linked to the dominant allele were identified and sequenced. These maps will serve as a starting point for future studies of the hazelnut genome, including map-based cloning of important genes. The inclusion of SSR loci on the map will make it useful in other populations.

  4. Construction of intersubspecific molecular genetic map of lentil based on ISSR, RAPD and SSR markers

    Indian Academy of Sciences (India)

    Mamta Gupta; Bhawna Verma; Naresh Kumar; Rakesh K. Chahota; Rajeev Rathour; Shyam K. Sharma; Sabhyata Bhatia; Tilak R. Sharma

    2012-12-01

    Lentil (Lens culinaris ssp. culinaris), is a self-pollinating diploid ($2n = 2x = 14$), cool-season legume crop and is consumed worldwide as a rich source of protein (∼24.0%), largely in vegetarian diets. Here we report development of a genetic linkage map of Lens using 114 F2 plants derived from the intersubspecific cross between L 830 and ILWL 77. RAPD (random amplified polymorphic DNA) primers revealed more polymorphism than ISSR (intersimple sequence repeat) and SSR (simple sequence repeat) markers. The highest proportion (30.72%) of segregation distortion was observed in RAPD markers. Of the 235 markers (34 SSR, 9 ISSR and 192 RAPD) used in the mapping study, 199 (28 SSRs, 9 ISSRs and 162 RAPDs) were mapped into 11 linkage groups (LGs), varying between 17.3 and 433.8 cM and covering 3843.4 cM, with an average marker spacing of 19.3 cM. Linkage analysis revealed nine major groups with 15 or more markers each and two small LGs with two markers each, and 36 unlinked markers. The study reported assigning of 11 new SSRs on the linkage map. Of the 66 markers with aberrant segregation, 14 were unlinked and the remaining 52 were mapped. ISSR and RAPD markers were found to be useful in map construction and saturation. The current map represents maximum coverage of lentil genome and could be used for identification of QTL regions linked to agronomic traits, and for marker-assisted selection in lentil.

  5. Genetic diversity among some canola cultivars as revealed by RAPD, SSR and AFLP analyses.

    Science.gov (United States)

    Moghaieb, Reda E A; Mohammed, Etr H K; Youssief, Sawsan S

    2014-08-01

    To assess the genetic diversity among four canola cultivars (namely, Serw-3, Serw-4, Misser L-16 and Semu 249), random amplified polymorphic DNA (RAPD), simple sequence repeat polymorphism (SSR) and amplified fragment length polymorphism (AFLP) analyses were performed. The data indicated that all of the three molecular markers gave different levels of polymorphism. A total of 118, 31 and 338 markers that show 61, 67.7 and 81 % polymorphism percentages were resulted from the RAPD, SSR and AFLP analyses, respectively. Based on the data obtained the three markers can be used to differentiate between the four canola cultivars. The genotype-specific markers were determined, 18 out of the 72 polymorphic RAPD markers generated were found to be genotype-specific (25 %). The highest number of RAPD specific markers was scored for Semu 249 (15 markers), while Serw-4 scored two markers. On the other hand, Serw-3 scored one marker. The cultivar Semu 249 scored the highest number of unique AFLP markers, giving 57 unique markers, followed by Misser L-16 which was characterized by 40 unique AFLP markers, then Serw-3 giving 31 unique markers. While Serw-4 was characterized by the lowest number producing 14 unique positive markers. The dendrogram built on the basis of combined data from RAPD, SSR and AFLP analysis represents the genetic distances among the four canola cultivars. Understanding the genetic variability among the current canola cultivars opens up a possibility for developing a molecular genetic map that will lead to the application of marker-assisted selection tools in genetic improvement of canola.

  6. RAPD and SSR Polymorphisms in Mutant Lines of Transgenic Wheat Mediated by Low Energy Ion Beam

    Institute of Scientific and Technical Information of China (English)

    WANG Tiegu; HUANG Qunce; FENG Weisen

    2007-01-01

    Two types of markers-random amplified polymorphic DNA (RAPD) and simple sequence repeat DNA (SSR)-have been used to characterize the genetic diversity among nine mutant lines of transgenic wheat intermediated by low energy ion beam and their four receptor cultivars. The objectives of this study were to analyze RAPD-based and SSR-based genetic variance among transgenic wheat lines and with their receptors, and to find specific genetic markers of special traits of transgenic wheat lines. 170 RAPD primers were amplified to 733 fragments in all the experimental materials. There were 121 polymorphic fragments out of the 733 fragments with a ratio of polymorphic fragments of 16.5%. 29 SSR primer pairs were amplified to 83 fragments in all the experiment materials. There were 57 polymorphic fragments out of the 83 fragments with a ratio of polymorphic fragments of 68.7%. The dendrograms were prepared based on a genetic distance matrix using the UPGMA (Unweighted Pair-group Method with Arithmetic averaging) algorithm, which corresponded well to the results of the wheat pedigree analysis and separated the 13 genotypes into four groups. Association analysis between RAPD and SSR markers with the special traits of transgenic wheat mutant lines discovered that three RAPD markers, si, opt-16, and fl4, were significantly associated with the muticate trait, while three SSR markers, Rht8 (Xgwm261), Rht-Blb, and Rht-Dlb, highly associated with the dwarf trait. These markers will be useful for marker-assistant breeding and can be used as candidate markers for further gene mapping and cloning.

  7. RAPD-PCR – still a suitable Method for Genetically Underexplored Species?

    Directory of Open Access Journals (Sweden)

    Konstanze Ursula Behrmann

    2015-11-01

    Full Text Available Saithe (Pollachius virens is a commercially important fish species; the annual catch quota in the Northeast Atlantic exceeds 100.000 t. Despite that saithe is underexplored from a fish population genetically view. Because saithe is a highly migratory species, which undergoes a long larval drift, the population structure of saithe within the Northeast Atlantic is not fully understood. Models used as a basis for the management plan are based on tagging studies, which have been carried out in the 1960th. But still there are doubts regarding the numbers of stocks living in the Northeast Atlantic. Migration routes are affected by salmon farming, growing steadily from the 1990th. In the last years a hyperstability of the saithe stock in the North Sea had been detected underlining the need to have a closer look on the saithe stocks in the Northeast Atlantic. Random amplified polymorphic DNA (RAPD - PCR is a DNA fingerprinting technique often used in species identification and population genetic research for species, whose genome has not been sequenced very extensive as being the case for most of the food fishes. We applied RAPD-PCR in a study of saithe populations from the North Atlantic. The suitability of RAPD-PCR was improved by optimisations for enhanced reproducibility. The “classical” protocol for RAPD-PCR was modified by increasing the annealing temperature and shortening the time of annealing, providing a much better reproducibility. Thus, RAPD-PCR was found to be a straightforward and low-cost way, compared to other population genetic tools, to get a first insight into the population structure of less sequenced fish species within a very short time, being useful for preliminary studies or laboratories without large capacities for DNA sequencing.

  8. Evaluation of genetic diversity in Piper spp using RAPD and SRAP markers.

    Science.gov (United States)

    Jiang, Y; Liu, J-P

    2011-11-29

    Random amplified polymorphic DNA (RAPD) and sequence-related amplified polymorphism (SRAP) analysis were applied to 74 individual plants of Piper spp in Hainan Island. The results showed that the SRAP technique may be more informative and more efficient and effective for studying genetic diversity of Piper spp than the RAPD technique. The overall level of genetic diversity among Piper spp in Hainan was relatively high, with the mean Shannon diversity index being 0.2822 and 0.2909, and the mean Nei's genetic diversity being 0.1880 and 0.1947, calculated with RAPD and SRAP data, respectively. The ranges of the genetic similarity coefficient were 0.486-0.991 and 0.520-1.000 for 74 individual plants of Piper spp (the mean genetic distance was 0.505 and 0.480) and the within-species genetic distance ranged from 0.063 to 0.291 and from 0.096 to 0.234, estimated with RAPD and SRAP data, respectively. These genetic indices indicated that these species are closely related genetically. The dendrogram generated with the RAPD markers was topologically different from the dendrogram based on SRAP markers, but the SRAP technique clearly distinguished all Piper spp from each other. Evaluation of genetic variation levels of six populations showed that the effective number of alleles, Nei's gene diversity and the Shannon information index within Jianfengling and Diaoluoshan populations are higher than those elsewhere; consequently conservation of wild resources of Piper in these two regions should have priority.

  9. Characterization of Leishmania Parasites Isolated From Kala- azar Patients in Kohgiloyeh and Boyerahmad, Using Semi-Nested PCR

    Directory of Open Access Journals (Sweden)

    B Sarkari

    2006-04-01

    Full Text Available ABSTRACT: Introduction & Objective: Visceral leishmaniasis (VL is a disease commonly known as Kala-azar caused by protozoan parasites of the genus Leishmania including L. donovani, L. infantum and L. chagasi. VL is sporadic in many areas of Iran and is endemic in a few provinces such as Fars, Azarbayjan, Bushehr, Ardabil and Qom. VL has been reported from some areas of Kohgiloyeh and Boyerahmad and this study aimed to characterize the causative agent of VL in this region. Materials & Methods: Bone marrow sample was obtained from 6 VL patients from children department in Imam Sajad hospital in Yasuj. DNA was extracted from the obtained samples and was checked by semi-nested PCR to determine the species of the parasite. To do that, a segment of minicircle kinetoplast DNA was amplified, using LINR4 and LIN17 primers. Products of PCR were evaluated by electrophoresis, using 1.5% agarose and stained with ethidium bromide. Results: Parasitologically examination of bone marrow smears demonstrated amastigotes form of the parasite in the samples. For mass cultivation, isolated parasites were cultured in diphasic NNN followed by RPMI 1640 media. All the samples produced a 720 bp band in PCR assay. The isolates were compared with referent strains and it was revealed that all the isolates were L. infantum. Conclusion: Findings of this study demonstrated that the causative agent of VL in Kohgiloyeh and Boyerahmad was L. infantum. Further study is needed to explore other aspects of VL in this region.

  10. Treatment-Based Strategy for the Management of Post-Kala-Azar Dermal Leishmaniasis Patients in the Sudan

    Directory of Open Access Journals (Sweden)

    A. M. Musa

    2013-01-01

    Full Text Available Post-kala-azar dermal leishmaniasis (PKDL is a dermatosis that affects more than 50% of successfully treated visceral leishmaniasis (VL patients in Sudan. PKDL is considered an important reservoir for the parasite and its treatment may help in the control of VL. Currently, treatment is mainly with sodium stibogluconate (SSG, an expensive and fairly toxic drug and without universally in treatment protocols used. A literature review, a consensus of a panel of experts, and unpublished data formed the basis for the development of guidelines for the treatment of PKDL in the Sudan. Six treatment modalities were evaluated. Experts were asked to justify their choices based on their experience regarding of drug safety, efficacy, availability, and cost. The consensus was defined by assigning a categorical rank (first line, second line, third line to each option. Regarding the use of AmBisome the presence of the drug in the skin was confirmed in smears from PKDL lesions. Recommendations: AmBisome at 2.5 mg/kg/day/20 days or SSG at 20 mg/kg/day/40 days plus four/weekly intradermal injection of alum-precipitated autoclave L. major vaccine are suggested as first- and second-treatment options for PKDL in the Sudan, respectively. SSG at 20 mg/Kg/day/60 or more days can be used if other options are not available.

  11. Choice of providers for treating a neglected tropical disease:an empirical analysis of kala azar in Nepal

    Institute of Scientific and Technical Information of China (English)

    ShivaRajAdhikari; SiripenSupakankunti; MMahmudKhan

    2011-01-01

    Objective:To examine the choice of healthcare providers for treating kala azar (KA) in Nepal. Methods:Information was collected from clinically diagnosed KA patients seeking care from public hospitals located in KA endemic districts. The survey collected information from more than 25 percent of total KA cases in the country. For empirical estimation of probability of choosing a provider-type as a first contact healthcare provider, a multinomial logit model was defined with five alternative options with self care as the reference category. Results: The empirical model found that price of medical care services, income of households, knowledge of patients on KA and KA treatment, borrowing money, age of patient, perceived quality of provider types, etc. determine the likelihood of seeking care from the alternative options considered in the analysis. All variables have expected signs and are consistent with earlier studies. The price and income elasticity were found to be very high indicating that poorer households are very sensitive to price and income changes, even for a severe disease like KA. Using the empirical models, we have analyzed two policy instruments:demand side financing and interventions to improve the knowledge index about KA. Conclusions:Due to high price elasticity of KA care and high spillover effects of KA on the society, policy makers may consider demand side financing as an instrument to encourage utilization of public hospitals.

  12. Discrimination Capacity of RAPD, ISSR and SSR Markers and of their Effectiveness in Establishing Genetic Relationship and Diversity among Egyptian and Saudi Wheat Cultivars

    Directory of Open Access Journals (Sweden)

    Salah E.D. El-Assal

    2012-01-01

    Full Text Available Problem statement: Yield crop cultivars and landraces are valuable sources of genetic variations that the knowledge and implication of these variations are critical in the plant breeding programs. our major objective of this study is investigating the discriminating capacity of RAPD, ISSR and SSR markers and of their effectiveness in establishing genetic relationship and diversity among Egyptian and Saudi wheat cultivars. Approach: Eleven wheat cultivars and landraces collected from Egypt and Saudi Arabia, five Egyptian wheat (Sakha 93, Sods 1, Sods 4, Gmiza 9 and Sohag 3 and six Saudi wheat landrace cultivars (Hmees, Al-Kaseem, Hegazi, Abo-Sakr, Dubai 1 and Nagran were characterized using RAPD, ISSR and SSR molecular markers as efficient tools. Ten and nine oligonucleotide primers of RAPD and ISSR respectively and four primer pairs of SSR were used in wheat samples analysis. Only clear and repeatable band profile of 6 RAPD, 8 ISSR and 2 SSR primers were obtained. In RAPD analyses, 74 out of 141 bands (52% were polymorphic. Results: The number of alleles ranged from 8-21 per primer, with an average of 14.1 per primer. In ISSR analyses, a total of 78 alleles were detected, along with 36 alleles (46% were polymorphic. The number of alleles per primer ranged from 5-10 with an average of 8.6 alleles per ISSR primer. SSR reactions recorded 6 alleles, of which 5 alleles (83% were polymorphic. Cluster analysis was conducted using Unweighted Pair Group Method that depends on Arithmetic Average (UPGMA. The dendrogram cluster diagram classified the evaluated genotypes in three major clusters corresponding to the cultivation regions. The first group contains Sakha 93, Sods 1 and Sods 4 with more than 80% Genetic Similarity (GS. The GS between Sakha 93 and Sods 1, Sakha 93 and Sods 4 or Sods 1 and Sods 4 were 83.6%, 83.9 and 85.4 respectively. The second group contains Gmiza 9 and Sohag 3 with GS 83.1%. The third group contains most of the Saudi landrace

  13. Post Kala-Azar Dermal Leishmaniasis following Treatment with 20 mg/kg Liposomal Amphotericin B (Ambisome) for Primary Visceral Leishmaniasis in Bihar, India

    OpenAIRE

    Sakib Burza; Prabhat Kumar Sinha; Raman Mahajan; Marta González Sanz; María Angeles Lima; Gaurab Mitra; Neena Verma; Pradeep Das

    2014-01-01

    BACKGROUND: The skin disorder Post Kala-Azar Dermal Leishmaniasis (PKDL) occurs in up to 10% of patients treated for visceral leishmaniasis (VL) in India. The pathogenesis of PKDL is not yet fully understood. Cases have been reported in India following therapy with most available treatments, but rarely in those treated with liposomal amphotericin B (Ambisome). Between July 2007 and August 2012 with the support of the Rajendra Memorial Research Institute (RMRI), Médecins Sans Frontières (MSF) ...

  14. The FML (Fucose Mannose Ligand of Leishmania donovani: a new tool in diagnosis, prognosis, transfusional control and vaccination against human kala-azar

    Directory of Open Access Journals (Sweden)

    Claris B. Palatnik de Sousa

    1996-04-01

    Full Text Available The Fucose-Mannose Ligand (FML of Leishmania donovani is a complex glycoproteic fraction. Its potential use as a tool for diagnosis of human visceral leishmaniasis was tested with human sera from Natal, Rio Grande do Norte, Brazil. The FML-ELISA test, showed 100% sensitivity and 96% specificity, identifying patients with overt kala-azar (p < 0.001, when compared to normal sera, and subjects with subclinical infection. More than 20% apparently healthy subjects with positive reaction to FML developed overt kala-azar during the following 10 months. In the screening of human blood donnors, a prevalence of 5% of sororeactive subjects was detected, attaining 17% in a single day. The GP36 glycoprotein of FHL is specifically reconized by human kala-azar sera. The immunoprotective effect of FML on experimental L. donovanii infection was tested in swiss albino mice. The protection scheemes included three weekly doses of FML, supplemented or not with saponin by the subcutaneous or intraperitoneal routes and challenge with 2x 10(7 amastigotes of Leishmania donovani. An enhancement of 80.0 % in antibody response (p<0.001 and reduction of 85.5 % parasite liver burden (p<0.001 was detected in animals immunized with FML saponin, unrespectivety of the immunization route.

  15. Analisis Keragaman Genetik Tanaman Aren (Arenga pinnata Merr) di Tapanuli Selatan dengan Menggunakan Marka RAPD (Random Amlpified Polymorphic DNA)

    OpenAIRE

    Harahap, Mahyuni Khairiyah

    2013-01-01

    MAHYUNI KHAIRIYAH HARAHAP : RAPD analysis on genetic diversity of sugar palm ( Arenga pinnata Merr) in South Tapanuli population. Supervised by LOLLIE AGUSTINA P. PUTRI and MOHAMMAD BASYUNI. The objective of this research was to analysis genetic diversity of natural sugar palm in South Tapanuli using Random Amplified Polymorphic DNA (RAPD). A total of 24 acessions palm sugar the populations originated from various regions in South Tapanuli, consists of West Angkola, South Angkola, Batan...

  16. Potential use of RAPD markers in characteristics of cucumber (Cucumis sativus L. haploids and double-haploids

    Directory of Open Access Journals (Sweden)

    Katarzyna Niemirowicz-Szczytt

    2011-04-01

    Full Text Available A study was designed to obtain hapolid and double haploid (DH plants from cucumber cultivars tolerant to Pseudoperonospora cubensis. The main goal was to identify RAPD molecular markers associated with downy mildew resistance. On average, 20% of embryos generated in two experiments, were converted to haploid plants. RAPD markers that differentiated susceptible and resistant H and DH plants were identified. Somaclonal variation in DH lines was seldom detected.

  17. Factores de riesgo asociados al embarazo ectópico

    OpenAIRE

    Jónathan Hernández Núñez; Magel Valdés Yong; Andrés Oscar Torres Chávez

    2013-01-01

    Fundamento: el embarazo ectópico se ha duplicado en la mayor parte de los países industrializados y representa una de las principales causas de muerte materna. Objetivo: identificar factores de riesgo que predisponen al embarazo ectópico.Métodos: estudio de casos y controles realizado en un universo de 132 pacientes del Municipio Habana del Este, operadas de embarazo ectópico en el Hospital Dr. Luís Díaz Soto desde el 2009 al 2011. Se escogió una muestra al azar de 66 pacientes que constituyó...

  18. RAPD and microsatellite transferability studies in selected species of Prosopis (section Algarobia) with emphasis on Prosopis juliflora and P. pallida

    Indian Academy of Sciences (India)

    Minu Sherry; Steve Smith; Ashok Patel; Phil Harris; Paul Hand; Liz Trenchard; Janey Henderson

    2011-08-01

    The genus Prosopis (Leguminosae, Mimosoideae), comprises 44 species widely distributed in arid and semi-arid zones. Prosopis pallida (Humb. & Bonpl. ex Willd.) Kunth and P. juliflora (Sw.) DC. are the two species that are truly tropical apart from P. africana, which is native to tropical Africa (Pasiecznik et al. 2004), and they have been introduced widely beyond their native ranges. However, taxonomic confusion within the genus has hampered exploitation and better management of the species. The present study focusses primarily on evaluating the genetic relationship between Prosopis species from the section Algarobia, containing most species of economic importance, though P. tamarugo from section Strombocarpa is also included for comparison. In total, 12 Prosopis species and a putative P. pallida × P. chilensis hybrid were assessed for their genetic relationships based on RAPD markers and microsatellite transferability. The results show that P. pallida and P. juliflora are not closely related despite some morphological similarity. Evidence also agrees with previous studies which suggest that the grouping of series in section Algarobia is artificial.

  19. RAPD and microsatellite transferability studies in selected species of Prosopis (section Algarobia) with emphasis on Prosopis juliflora and P. pallida.

    Science.gov (United States)

    Sherry, Minu; Smith, Steve; Patel, Ashok; Harris, Phil; Hand, Paul; Trenchard, Liz; Henderson, Janey

    2011-08-01

    The genus Prosopis (Leguminosae, Mimosoideae), comprises 44 species widely distributed in arid and semi-arid zones. Prosopis pallida (Humb. and Bonpl. ex Willd.) Kunth and P. juliflora (Sw.) DC. are the two species that are truly tropical apart from P. africana, which is native to tropical Africa (Pasiecznik et al. 2004), and they have been introduced widely beyond their native ranges. However, taxonomic confusion within the genus has hampered exploitation and better management of the species. The present study focusses primarily on evaluating the genetic relationship between Prosopis species from the section Algarobia, containing most species of economic importance, though P. tamarugo from section Strombocarpa is also included for comparison. In total, 12 Prosopis species and a putative P. pallida x P. chilensis hybrid were assessed for their genetic relationships based on RAPD markers and microsatellite transferability. The results show that P. pallida and P. juliflora are not closely related despite some morphological similarity. Evidence also agrees with previous studies which suggest that the grouping of series in section Algarobia is artificial.

  20. Diversidade genética de três estoques de piapara (Leporinus elongatus, utilizando RAPD = Genetic diversity of three stocks of piapara (Leporinus elongatus, using RAPD

    Directory of Open Access Journals (Sweden)

    Patrícia Cristina Gomes

    2008-04-01

    Full Text Available Recentemente a produção aquícola brasileira tem apresentado grandeprogresso. Dentre as espécies nativas cultivadas no Brasil, a piapara (Leporinus elongatus tem sido amplamente preconizada. Com objetivo de avaliar os programas de repovoamento, foram analisadas a variabilidade e a divergência genética de três estoques de piapara com a técnica de RAPD (Random Amplified Polymorphic. O primeiro estoque pertence à Estação de Aquicultura e Hidrologia da Duke Energy International (A; o segundo, à piscicultura de Rolândia (B e o terceiro, ao Programa de Repovoamento dos Rios do Paraná (C. Os dezprimers para RAPD utilizados produziram 105 fragmentos polimórficos, conferindo um polimorfismo de 98,1% para os três estoques avaliados. A porcentagem de locos polimórficos e índice de Shannon foi superior para o estoque A. Porém, todos valores foram elevados, indicando alta diversidade intrapopulacional. Os valores de Gst indicam que houvebaixa diferenciação genética entre os estoques A x B e moderada diferenciação entre os demais. O Nm foi maior entre os estoques A x B. A distância genética e o dendrograma indicam que os estoques A x B são menos distantes geneticamente.Latelly, aquiculture production in Brazil has made great strides. Among the native species cultivated in Brazil, piapara (Leporinus elogatus has been widely praised. With the objective of evaluating restocking programs, the variability and genetic divergence ofthree piapara stocks were analyzed using the RAPD (Random Amplified Polymorphic DNA technique. The first stock belongs to the Aquiculture and Hydrology Station of Duke Energy International (A; the second one belongs to a fish farm in the city of Rolândia(B; and the third to the River Restocking Program of Paraná (C. The ten primers used for RAPD produced 105 polymorphic loci, conferring a polymorphism of 98.1% for the three evaluated stocks. Polymorphic loci percentage and Shannon index were higher for stock A

  1. Genetic diversity of Phytophthora sojae isolates in Heilongjiang Province in China assessed by RAPD and EST-SSR

    Science.gov (United States)

    Wu, J. J.; Xu, P. F.; Liu, L. J.; Wang, J. S.; Lin, W. G.; Zhang, S. Z.; Wei, L.

    Random-amplified polymorphic DNA (RAPD) and EST-SSR markers were used to estimate the genetic relationship among thirty-nine P.sojae isolates from three locations in Heilongjiang Province, and nine isolates from Ohio in America were made as reference strains. 10 of 50 RAPD primers and 5 of 33 EST-SSR were polymorphic across 48 P.sojae isolates. Similarity values among P.sojae isolates were from 49% to 82% based on the RAPD data. The similarities based on EST-SSR markers ranged from 47% to 85%. The genetic diversity revealed by EST-SSR marker analysis was higher than that obtained from RAPD. The similarity matrices for the SSR data and the RAPD data were moderately correlated (r = 0.47). Genetic similarity coefficients were also relatively lower, which demonstrated complicated genetic background within each location. The high similarity values range revealed the ability of RAPD/EST-SSR markers to distinguish even among morphological similar phytophthora.

  2. Genetic characterization of early maturing maize hybrids (Zea mays L. obtained by protein and RAPD markers

    Directory of Open Access Journals (Sweden)

    Bauer Iva

    2005-01-01

    Full Text Available Knowledge of maize germplasm genetic diversity is important for planning breeding programmes, germplasm conservation per se etc. Genetic variability of maize hybrids grown in the fields is also very important because genetic uniformity implies risks of genetic vulnerability to stress factors and can cause great losts in yield. Early maturing maize hybrids are characterized by shorter vegetation period and they are grown in areas with shorter vegetation season. Because of different climatic conditions in these areas lines and hybrids are developed with different features in respect to drought resistance and disease resistance. The objective of our study was to characterize set of early maturing maize hybrids with protein and RAPD markers and to compare this clasification with their pedigree information. RAPD markers gave significantly higher rate of polymorphism than protein markers. Better corelation was found among pedigree information and protein markers.

  3. Molecular markers of nuclear restoration gene Rf1 in sunflower using bulked segregant analysis-RAPD

    Institute of Scientific and Technical Information of China (English)

    季静; 王罡; E.Belhassen; H.Serieys; A.Berville

    1996-01-01

    Restoration of cytoplasmic male sterility (CMS) in sunflower was demonstrated to be controlled by polygenes by analysing 982 effective crosses among 109 self-crossed lines and 16 CMS lines. Two self-crossed lines and one CMS line with distinct genotypes were applied to creation of segregating populations for DNA bulks of the target gene Rfl. Bulked DNA was prepared in order to investigate single gene Rfl and its gene marker among polygenic characters at the same genetic background. Using 80 10-mer operon primers, 620 RAPD reactions were carried out between fertile and sterile DNA bulks. In about 800 loci, primary results showed that 8 were related to the restoration genes. Furthermore. 2 were confirmed as RAPD markers for gene Rfl by examining 9 maintenance and 7 restoration lines. This method is the improvement for bulked segregant analysis[1] with which markers of single gene of target can be identified rapidly among polygenic characters.

  4. Genetic diversity of alfalfa domesticated varietal populations from Libyan genbank revealed by RAPD markers

    Directory of Open Access Journals (Sweden)

    Ahsyee Salem R.

    2013-01-01

    Full Text Available Alfalfa (Medicago sativa L. is an important forage legume in Libya. The genetic diversity of nine alfalfa domesticated varietal populations was studied using thirteen RAPD primer combinations. The number of polymorphic fragments detected per primer combination ranged from 8 to 46 bands with an average of 24 bands. The number of polymorphic bands detected was from 6 (Atalia population to 37 (Gabsia population. The lowest genetic distance was 0.058 and the highest was 0.655. The average genetic distance was (0.356. The dendrogram based on Ward’s minimum variance clustering method grouped the nine populations into the two main clusters. The first group included Fazania, Atalia, Masratia, Zawia, Denamo Ferade and Arezona. The second group was composed of Tagoria, Gabsia and Wade Alrabeh. The simplicity of RAPD assays for detection of genetic polymorphisms is confirmed in our study, and results can be utilized in breeding practice.

  5. Molecular Polymorphisms in Tunisian Pomegranate (Punica granatum L. as Revealed by RAPD Fingerprints

    Directory of Open Access Journals (Sweden)

    Jemni Chibani

    2010-01-01

    Full Text Available The genetic diversity among Tunisian pomegranate cultivars has been investigated. Using universal primers, the random amplified polymorphic DNA (RAPD method was used to generate banding profiles from a set of twelve cultivars. Data was then computed with appropriate programs to construct a dendrogram illustrating the relationships between the studied cultivars. Our data proved the efficiency of the designed method to examine the DNA polymorphism in this crop since the tested primers are characterized by a collective resolving power of 12.83. In addition, the cluster analysis has exhibited a parsimonious tree branching independent from the geographic origin of the cultivars. In spite of the relatively low number of primers and cultivars, RAPD constitutes an appropriate procedure to assess the genetic diversity and to survey the phylogenetic relationships in this crop.

  6. Genetic relationships among Chinese and American isolates of Phytophthora sojae assessed by RAPD markers

    Institute of Scientific and Technical Information of China (English)

    WANG Ziying; WANG Yuanchao; ZHANG Zhengguang; ZHENG Xiaobuo

    2006-01-01

    The genetic diversity of three geographic populations of Phytophthora sojae from China and the United States was determined using random amplified polymorphic DNA (RAPD). The purpose was to explore genetic relationships among Chinese and American isolates of the organism. 21 random primers were selected among 200 random primers screened. A total of 223 reproducible RAPD fragments were scored among 111 individuals, of which 199 (89.23%) were polymorphic. Analysis of genetic variation showed that there existed higher genetic variation in the United States population in comparison to the Chinese populations. Nei's genetic identity and principal component analysis indicated that the populations of Fujian and United States are closer to each other than to Heilongjiang populations. Shannon-Wiener diversity index revealed that the United States populations have a higher genetic diversity than that of Chinese populations. These data are in support of the hypothesis that P. Sojae in the United States might not have been introduced from China.

  7. Combination of ARDRA and RAPD genotyping techniques in identification of Acinetobacter spp. genomic species

    Institute of Scientific and Technical Information of China (English)

    Yong ZHANG; Yuqing CHEN; Yingchun TANG; Kouxing ZHANG

    2008-01-01

    A total of 10 non-repetitive multi-drug-resist-ant Acinetobacter strains were collected. With reference to A. calcoaceticus (ATCC23055), A. baumannii (ATCC19606), A. lwoffii (ATCC17986), and A. junii (NCTC5866), DNA fingerprint technique, amplified ribo-somal DNA restriction analysis (ARDRA), and random amplified polymorphism DNA (RAPD) were carried out to identify the genomic species of Acinetobacter spp. The distances between them were calculated by the unweighted pair group method with arithmetic (UPGMA). Genotypes ofAcinetobacter spp. were effectively classified and an A. junii together with nine A. baumannii isolates was genomically identified. The combination of ARDRA and RAPD DNA-fingerprint technique shows high com-plementarity, and could be a useful tool in Acinetobacter genomic species identification.

  8. Genetic diversity revealed in commercial varieties of Chrysanthemum (Dendranthemagrandiflora using RAPD markers

    Directory of Open Access Journals (Sweden)

    P.Lalitha Kameswari1, G.Anuradha2 , M. Pratap3and Hameedunnisabegum4

    2014-03-01

    Full Text Available Molecular characterization using RAPD analysis was carried out in 37 chrysanthemum genotypes.With 27 RAPD primers, total of 278 amplified fragments were scored.Out of which 271 were found to be polymorphic (97.4%. The genetic similarity ranged from a coefficient of 0.174 to 0.600 with an average similarity coefficient of 0.387 indicating a moderate diversity among the group of genotypes studied. In the present study, Ratlam Selection and Snow Cem were found to be the most genetically similar (0.60 followed by Akitha and Shintome with 55.3%. Cluster analysis based on UPGMA method led to the classification of genotypes intofour major clusters and seven minor clusters with one genotype each in cluster V (Arka Ravi, VI (Meera, VII(Asha, VIII(Silper, IX(Autumn Joy, X (Lilith and cluster XI (Chandrika.

  9. Haploid Origin of Cork Oak Anther Embryos Detected by Enzyme and RAPD Gene Markers.

    Science.gov (United States)

    Bueno; Agundez; Gomez; Carrascosa; Manzanera

    2000-05-01

    In vitro-induced cork oak (Quercus suber L.) embryos from anther cultures proved to be of haploid origin both by enzyme and RAPD gene marker analysis. The problem considered was to ascertain if embryo cultures originated either from a single haploid cell, from a microspore, or from multiple haploid cells. Therefore, a heterozygotic gene was searched for in the parent tree. The gene coding for shikimate dehydrogenase (SKDH1) proved to be heterozygous in the parental tree, and subsequently, these allozymes were screened for the embryos induced in anther cultures from the same tree. Only haploid embryos were found, confirming the microspore origin. Different genotypes were not identified inside each anther by isozyme analysis, probably because of selective pressure for one embryo early in development, but both parental SKDH1 alleles were found in the embryos of different anthers. The banding patterns detected by RAPD markers permitted the identification of multiple microspore origins inside each anther.

  10. Genetic Diversity Analysis of Iranian Improved Rice Cultivars through RAPD Markers

    Directory of Open Access Journals (Sweden)

    Ghaffar KIANI

    2011-08-01

    Full Text Available The aim of this study was to evaluate the genetic diversity of Iranian improved rice varieties. Sixteen rice varieties of particular interest to breeding programs were evaluated by means of random amplified polymorphic DNA (RAPD technique. The number of amplification products generated by each primer varied from 4 (OPB-04 to 11 (OPD-11 with an average of 8.2 bands per primer. Out of 49 bands, 33 (67.35% were found to be polymorphic for one or more cultivars ranging from 4 to 9 fragments per primer. The size of amplified fragments ranged between 350 to 1800 bp. Pair-wise Nei and Li�s (1979 similarity estimated the range of 0.59 to 0.98 between rice cultivars. Results illustrate the potential of RAPD markers to distinguish improved cultivars at DNA level. The information will facilitate selection of genotypes to serve as parents for effective rice breeding programs in Iran.

  11. IDENTIFICATION OF RAPD MARKERS LINKED TO MYMV RESISTANCE IN MUNGBEAN (VIGNA RADIATA (L). WILCZEK)

    OpenAIRE

    PRAVEEN HOLEYACHI; D L SAVITHRAMMA

    2013-01-01

    The present study was undertaken to identify random amplified polymorphic DNA (RAPD) marker associated with mungbean yellow mosaic virus (MYMV) resistance in mungbean (Vigna radiata (L). Wilczek) by employing bulk segregant analysis in recombinant inbred lines (RILs). Out of 20 random decamers, only ten primers viz; OPA-03, A-06, A-03, OPA-09, A-09, OPB 7, UBC-391, OPC-08, UBC 499 and A-04 showed polymorphism between parents Chinamung and BL 849. Out of these ten primers only one ...

  12. Effect of nickel on regeneration in Jatropha curcas L. and assessment of genotoxicity using RAPD markers

    KAUST Repository

    Sarkar, Tanmoy

    2010-07-08

    The aim of the present study was to determine the effect of nickel on shoot regeneration in tissue culture as well as to identify polymorphisms induced in leaf explants exposed to nickel through random amplified polymorphic DNA (RAPD). In vitro leaf explants of Jatropha curcas were grown in nickel amended Murashige and Skoog (MS) medium at four different concentrations (0, 0.01, 0.1, 1 mM) for 3 weeks. Percent regeneration, number of shoots produced and genotoxic effects were evaluated by RAPD using leaf explants obtained from the first three treatments following 5 weeks of their subsequent subculture in metal free MS medium. Percent regeneration decreased with increase in addition of nickel to the medium up to 14 days from 42.31% in control to zero in 1.0 mM. The number of shoot buds scored after 5 weeks was higher in control as compared to all other treatments except in one of the metal free subculture medium wherein the shoot number was higher in 0.01 mM treatment (mean = 7.80) than control (mean = 7.60). RAPD analysis produced only 5 polymorphic bands (3.225%) out of a total of 155 bands from 18 selected primers. Only three primers OPK-19, OPP-2, OPN-08 produced polymorphic bands. The dendrogram showed three groups A, B, and C. Group A samples showed 100% genetic similarity within them. Samples between groups B and C were more genetically distant from each other as compared to samples between groups A and B as well as groups A and C. Cluster analysis based on RAPD data correlated with treatments. © 2010 Springer Science+Business Media, LLC.

  13. Analysis of genetic relationship among Arbutus unedo L.genotypes using RAPD and SSR markers

    Institute of Scientific and Technical Information of China (English)

    Filomena Gomes; Rita Costa; Maria M.Ribeiro; Elisa Figueiredo; Jorge M.Canhoto

    2013-01-01

    The strawberry tree (Arbutus unedo L.) is an underutilized,drought tolerant,fire resistant species with a south western distribution in Europe,and with ecological and putative socio-economical impact in Portugal and Mediterranean countries.Our aim was to develop an appropriate set of molecular markers to enable genetic diversity to be assessed and to fingerprint Arbutus unedo genotypes for breeding and conservation purposes in Portugal.Twenty-seven trees from a broad geographic range were screened with 20 random amplified polymorphic DNA(RAPD primers) and 11 microsatellite markers (SSR).The RAPDs generated 124 bands,57.3% of which were polymorphic,with an expected heterozygosity of 27%.We cross-amplified 11 SSR primers developed for Vaccinium spp.,and 5 were found to be polymorphic in A.unedo,with 75% of expected heterozygosity,a number of alleles of 11.6,a null allele frequency of 7.6% and a polymorphic information content of 71%.Although the SSRs were more polymorphic and informative than the RAPDs,both markers displayed high genetic variability with the gathered data.No geographic pattern was observed in the genetic variation distribution based on both marker systems,and the lack of correlation between genetic and geographical matrices was confirmed by Mantel tests.Likely,no correlation was found between pairwise SSR and RAPD band-sharing matrices.These results and their implications on A.unedo breeding and conservation programs are discussed.

  14. Study on Phylogenetic Relationships of Five Breeds of Pigs by Random Amplified Polymorphic DNA(RAPD)

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    RAPD was used to study the genetic divergency and phylogenetic relationships of five breeds of domestic pigs,including Min pig,Duroc,Yorkshired,Landrace and Junmu I pig.We selected fourteen primers from eighty random primers,caculated genetic distance index matrix and constructed phylogenetic tree with UPGMA methods.Genetic distance index matrix indicated that the genetic relationship between Junmu I pig and Landrace was the closest and the farthest between Duroc and min pig.

  15. Diversity analysis of Moroccan carob ("Ceratonia siliqua" L.) accessions using phenotypic traits and RAPD markers

    OpenAIRE

    2007-01-01

    Diversity analysis of moroccan carob (Ceratonia siliqua L.) accessions using phenotypic traits and RAPD markers. The carob (Ceratonia siliqua L.) is a perennial leguminous (Caesalpinioideae) that grows as an evergreen shrub or tree. It¿s an important component of the Mediterranean vegetation and its adaptation in marginal soils of the Mediterranean regions is important environmentally and economically. Phenotypic and genetic diversity among 10 Ceratonia siliqua accessions coming from differen...

  16. RAPD Technique Used to Determine the Purity of Hybrid Hot Pepper Seeds

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Two hybrid hot pepper varieties Xiangyan 5 and Xiangyan 10, and their parents were analyzed the polymerase chain reaction with MJ /PT 200 Peltrier Themal Cycler and DS 800 White-ultravilot Transilluminator to set up a RAPD system adaptable to the purity determination of the hybrid seeds. Among the 39 random primers, 2 and 4 primers were found to be used effectively in Xiangyan 5 and Xiangyan 10 respectively.

  17. Salt Tolerance Evaluation in Cotton (Gossypium hirsutum L. Using RAPD Marker

    Directory of Open Access Journals (Sweden)

    Basel Saleh

    2015-12-01

    Full Text Available The aim of this study was to evaluate four upland cotton (Gossypium hirsutum L. [Niab78 (N78, Deir-Ezzor22 (DE22, Deltapine50 (DP50 and Aleppo118 (A118] varieties towards salt stress (0 and 200 mM NaCl for 7 weeks based on RAPD marker. Our data showed that the highest total polymorphic bands identified by the 26 RAPD tested primers were 150 bands generated by N78 variety, while the lowest ones were recorded for DP50 (29 bands. Otherwise, unique (negative and positive markers characterized the two tolerant varieties (N78 and DE22 were 22 and 29 markers, respectively. Our data indicated that the highest polymorphism level was detected in N78 variety (68.5% followed by DE22 (60.9%, whereas, the lowest one was recorded for DP50 (21.3%. Our data obtained herein indicates that RAPD marker provided molecular markers for salinity tolerance screening in early stage in genetic improvement programs.

  18. Genetic Diversity Analysis of Iranian Jujube Ecotypes (Ziziphus spp. Using RAPD Molecular Marker

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    S Abbasi

    2012-12-01

    Full Text Available Jujube (Ziziphus jujuba Mill. is a valuable medicinal plant which is important in Iranian traditional medicines. Although the regional plants such as jujube play an important role in our economy, but they are forgotten in research and technology. Considering the economic and medicinal importance of jujube, the first step in breeding programs is determination of the genetic diversity among the individuals. 34 ecotypes of jujube, which have been collected from eight provinces of Iran, were used in this study. The genetic relationships of Iranian jujube ecotypes were analyzed using Random Amplified Polymorphic DNA (RAPD marker. Six out of 15 random decamer primers applied for RAPD analysis, showed an informative polymorphism. According to clustering analysis using UPGMA's methods, the ecotypes were classified into two major groups at the 0.81 level of genetic similarity. The highest value of similarity coefficient (0.92 was detected between Mazandaran and Golestan ecotypes and the most genetic diversity was observed in ecotypes of Khorasan-Jonoubi. The affinity of Khorasan-Jonoubi and Esfahan ecotypes indicated a possible common origin for the variation in these areas. Results indicated that RAPD analysis could be successfully used for the estimation of genetic diversity among Ziziphus ecotypes and it can be useful for further investigations.

  19. Impact of gamma rays on the Phaffia rhodozyma genome revealed by RAPD-PCR

    Science.gov (United States)

    Najafi, N; Hosseini, Ramin; Ahmadi, AR

    2011-01-01

    Background and Objectives Phaffia rhodozyma is a red yeast which produces astaxanthin as the major carotenoid pigment. Astaxanthin is thought to reduce the incidence of cancer and degenerative diseases in man. It also enhances the immune response and acts as a free-radical quencher, a precursor of vitamin A, or a pigment involved in the visual attraction of animals as mating partners. The impact of gamma irradiation was studied on the Phaffia rhodozyma genome. Materials and Methods Ten mutant strains, designated Gam1-Gam10, were obtained using gamma irradiation. Ten decamer random amplified polymorphic DNA (RAPD) primers were employed to assess genetic changes. Results Nine primers revealed scorable polymorphisms and a total of 95 band positions were scored; amongst which 38 bands (37.5%) were polymorphic. Primer F with 3 bands and primer J20 with 13 bands produced the lowest and the highest number of bands, respectively. Primer A16 produced the highest number of polymorphic bands (70% polymorphism) and primer F showed the lowest number of polymorphic bands (0% polymorphism). Genetic distances were calculated using Jaccard's coefficient and the UPGMA method. A dendrogram was created using SPSS (version 11.5) and the strains were clustered into four groups. Conclusion RAPD markers could distinguish between the parental and the mutant strains of P. rhodozyma. RAPD technique showed that some changes had occurred in the genome of the mutated strains. This technique demonstrated the capability to differentiate between the parental and the mutant strains. PMID:22530091

  20. Assessment of Genetic Variation and Distribution Pattern of Thalictrum petaloideum Detected by RAPDs

    Institute of Scientific and Technical Information of China (English)

    XIELei; LILiang—Qian; ZHANGDa—Ming

    2004-01-01

    Random amplified polymerphic DNA(RAPD)method was applied to assessg enetic variation and population structure of Thahctrum petalotdeum L(Ranunoulaceae),Two hundred and forty-six individuals from 11 populations of the species were investigated by RAPD profiles Twenty selected RAPD primers generated 125 bands.in which 120 were polymorphic Ther esults revealed a high level of genetic variation(ercentage of polymorphIc bands(PPB was 96%.Nei’s gene diversity(りwas 03502 and shannon’s information index(I) was 0.5199 at the species level) The differentiation among the populations was high(Gst=0.3511)in this species.Result of analyzing of molecularvariance(AMOVA)showedthat38.88%of genetic variance was found among the populations Positive correlation withr r=01945(P=00002)was found between genetic distance and geographic distance amongpo pulations Two populations distributed in the drainage basin of YanELz River affined genedcally and formed one clada and the rest nine populations formed the other clade in both unweighted pair-group method using arithmetic average(UPGMA)trees made by two different method different methods. It was yen/clear that these two populations were very special, andmust be closely related in history, despite the fact that they now share quite weak link to the restpopulations through gene communication.

  1. Antibiotic resistance profile and RAPD analysis of Campylobacter jejuni isolated from vegetables farms and retail markets

    Institute of Scientific and Technical Information of China (English)

    John Yew Huat Tang; Mohd Ikhsan Khalid; Syazana Aimi; Che Abdullah Abu-Bakar; Son Radu

    2016-01-01

    Objective: To investigate antibiotic resistance profile and characterize Campylobacter jejuni (C. jejuni) isolates using random amplified polymorphic DNA (RAPD) analysis. Methods: Ninety eight C. jejuni isolates from farms and retail outlets were screened against 10 antibiotics commonly used clinically and agriculturally by using disk diffusion method. RAPD analysis was done to characterize 98 C. jejuni isolates. Results: Fifty-one percent of the isolates had multiple antibiotic resistance index 0.2 and below. This indicated that the isolates in the vegetables were not from the high risk environment or extensive farming practices. C. jejuni isolates found resistant towards penicillin G (93%), vancomycin (86%), ampicillin (35%), erythromycin (28%), genta-mycin (4%), amikacin (3%), enrofloxacin (1%), norfloxacin (1%) and no resistance to-wards ciprofloxacin. RAPD clustering analysis showed that the contamination of C. jejuni in vegetables was likely due to cross contamination at retail markets. Conclusions: C. jejuni contamination in vegetables at retail markets was due to cross contamination. Current finding proved that C. jejuni in small scale vegetables production was less expose towards antibiotic abuse.

  2. Antibiotic resistance profile and RAPD analysis of Campylobacter jejuni isolated from vegetables farms and retail markets

    Institute of Scientific and Technical Information of China (English)

    John Yew Huat Tang; Mohd Ikhsan Khalid; Syazana Aimi; Che Abdullah Abu-Bakar; Son Radu

    2016-01-01

    Objective:To investigate antibiotic resistance profile and characterize Campylobacter jejuni(C.jejuni) isolates using random amplified polymorphic DNA(RAPD) analysis.Methods:Ninety eight C.jejuni isolates from farms and retail outlets were screened against 10 antibiotics commonly used clinically and agriculturally by using disk diffusion method.RAPD analysis was done to characterize 98 C.jejuni isolates.Results:Fifty-one percent of the isolates had multiple antibiotic resistance index 0.2 and below.This indicated that the isolates in the vegetables were not from the high risk environment or extensive farming practices.C.jejuni isolates found resistant towards penicillin G(93%),vancomycin(86%),ampicillin(35%),erythromycin(28%),gentamycin(4%),amikacin(3%),enrofloxacin(1%),norfloxacin(1%) and no resistance towards ciprofloxacin.RAPD clustering analysis showed that the contamination of C.jejuni in vegetables was likely due to cross contamination at retail markets.Conclusions:C.jejuni contamination in vegetables at retail markets was due to cross contamination.Current finding proved that C.jejuni in small scale vegetables production was less expose towards antibiotic abuse.

  3. RAPD Assessment of Genetic Diversity of Yunjie(Eruca sativa Mill.) in China

    Institute of Scientific and Technical Information of China (English)

    SUN Wan-cang; WANG He-lin; GUAN Chun-yun; MENG Ya-xiong; ZHANG Jin-wen; LIU Zi-gang; ZHANG Tao; LI Xun; CHEN She-yuan; ZENG Xiu-cong

    2003-01-01

    Genetic diversity of Yunjie (Eruca sativa Mill. ) in China was assessed by analyses of RAPD (randomly amplified polymorphic DNA) markers. Twenty native cultivars representing Yunjie-growing ecotypes in China were selected as material in this study. Twelve out of the 64 tested random decamer primers were able to identify 131 stable RAPD bands from these Yunjie cultivars. Of them 105 bands, or 80.15% of the total, were polymorphic. Most Yunjie cultivars from the same ecotype had their characteristic DNA bands.Cluster analysis by unweighted pair group method of arithmetic means (UPGMA) suggested that the 20 Yunjie genotypes could be divided into four groups. The genetic distances among the 20 cultivars varied from 0. 117 8between Shuozhou and Shenchi to 0. 499 4 between Hetian and Xiliang. Hetian alone could be a new type of Yunjie identified in China because it had the greatest genetic distance from all the other tested cultivars. These results indicate that Chinese Yunjie have abundant genetic diversity. Classification of Chinese Yunjie based on the RAPD information was in good agreement with the relationships between these Yunjie cultivars in their geographic origins and their plant morphology.

  4. OPTIMALISASI EKSTRAKSI DNA DAN PCR-RAPD PADA GREVILLEA SPP. (PROTEACEAE

    Directory of Open Access Journals (Sweden)

    MADE PHARMAWATI

    2014-04-01

    Full Text Available Molecular genetic analysis of plants relies on high yield and high purity of DNA as well as optimized condition of molecular reactions. Appropriate methods for DNA extraction and molecular reactions such as PCR are therefore needed. This study aimed to develop protocol for extraction of high molecular weight DNA from Grevillea leaf and to optimize condition of PCR-RAPD. Standard plant DNA extraction of Doyle and Doyle was modified by increasing EDTA concentration to 50 mM and addition of 2% (v/v 2-mercaptoethanol. Moreover, incubation time was prolonged to 14-16 h at 55oC. This method yielded good quality of DNA and consistent results. Amplification of DNA using PCR-RAPD will become efficient and consistent if the amplification reactions are in ideal condition. In Grevillea, clear, reproducible and scorable PCR-RAPD patterns were obtained using 10ng DNA template, 5 pmol primer, 2.5 mM MgCl2 and the number of thermal cycle was 40 x.

  5. Typing of bacteriophages by randomly amplified polymorphic DNA (RAPD)-PCR to assess genetic diversity.

    Science.gov (United States)

    Gutiérrez, Diana; Martín-Platero, Antonio M; Rodríguez, Ana; Martínez-Bueno, Manuel; García, Pilar; Martínez, Beatriz

    2011-09-01

    The recent boom in phage therapy and phage biocontrol requires the design of suitable cocktails of genetically different bacteriophages. The current methods for typing phages need significant quantities of purified DNA, may require a priori genetic information and are cost and time consuming. We have evaluated the randomly amplified polymorphic DNA (RAPD)-PCR technique to produce unique and reproducible band patterns from 26 different bacteriophages infecting Staphylococcus epidermidis, Staphylococcus aureus, Lactococcus lactis, Escherichia coli, Streptococcus thermophilus, Bacillus subtilis and Lactobacillus casei bacterial strains. Initially, purified DNA and phage suspensions of seven selected phages were used as a template. The conditions that were found to be optimal 8 μM of 10-mer primers, 3 μM magnesium oxalacetate and 5% dimethyl sulfoxide. The RAPD genomic fingerprints using a phage titer suspension higher than 10(9) PFU mL(-1) were highly reproducible. Clustering by the Pearson correlation coefficient and the unweighted pair group method with arithmetic averages clustering algorithm correlated largely with genetically different phages infecting the same bacterial species, although closely related phages with a similar DNA restriction pattern were indistinguishable. The results support the use of RAPD-PCR for quick typing of phage isolates and preliminary assessment of their genetic diversity bypassing tedious DNA purification protocols and previous knowledge of their sequence.

  6. Genetic relationships among some Pinus, Picea and Abies species revealed by RAPD markers

    Directory of Open Access Journals (Sweden)

    Kovačević Dragan

    2013-01-01

    Full Text Available Studies were undertaken to identify genetic relationships among ten different species of the family Pinaceae through randomly amplified polymorphic DNA (RAPD markers. Eighteen arbitrary RAPD primers produced 123 fragments of which 107 were polymorphic (87%. The similarity coefficient values varied from 0.34 to 0.67. The highest similarity coefficient was detected between Pinus wallichiana and P. strobus as well as between Picea abies and P. orientalis, and the lowest was detected between threePinus species (P. heldreichii, P. peuce and P. wallichiana and Picea omorika. The analysis of RAPD markers confirmed the genetic relationships among species. GenusPicea is clearly separated from genus Pinus and is closer to genus Abies (A. concolor than to genus Pinus, what confirms up-to-date numerous comparative-morphological, anatomical, chemotaxonomic and molecular results of these closely related genera. Furthermore, on the basis of our results, pine species from different subgenera -Pinus and Strobus are clearly separated. This statement is in agreement with contemporary intrageneric classification of the genus Pinus. [Projekat Ministarstva nauke Republike Srbije, br.173029

  7. Application of RAPD for molecular characterization of plant species of medicinal value from an arid environment.

    Science.gov (United States)

    Arif, I A; Bakir, M A; Khan, H A; Al Farhan, A H; Al Homaidan, A A; Bahkali, A H; Al Sadoon, M; Shobrak, M

    2010-11-09

    The use of highly discriminatory methods for the identification and characterization of genotypes is essential for plant protection and appropriate use. We utilized the RAPD method for the genetic fingerprinting of 11 plant species of desert origin (seven with known medicinal value). Andrachne telephioides, Zilla spinosa, Caylusea hexagyna, Achillea fragrantissima, Lycium shawii, Moricandia sinaica, Rumex vesicarius, Bassia eriophora, Zygophyllum propinquum subsp migahidii, Withania somnifera, and Sonchus oleraceus were collected from various areas of Saudi Arabia. The five primers used were able to amplify the DNA from all the plant species. The amplified products of the RAPD profiles ranged from 307 to 1772 bp. A total of 164 bands were observed for 11 plant species, using five primers. The number of well-defined and major bands for a single plant species for a single primer ranged from 1 to 10. The highest pair-wise similarities (0.32) were observed between A. fragrantissima and L. shawii, when five primers were combined. The lowest similarities (0) were observed between A. telephioides and Z. spinosa; Z. spinosa and B. eriophora; B. eriophora and Z. propinquum. In conclusion, the RAPD method successfully discriminates among all the plant species, therefore providing an easy and rapid tool for identification, conservation and sustainable use of these plants.

  8. [PCR-RAPD typing of carbapenem-resistant Pseudomonas aeruginosa strains].

    Science.gov (United States)

    Bogiel, Tomasz; Gospodarek, Eugenia

    2010-01-01

    P. aeruginosa rods are opportunistic pathogens responsible generally for nosocomial infections. Resistance to carbapenems, observed among them, is a serious threat due to ability to be transmitted between bacterial species. The aim of our study was to evaluate the usefulness of PCR-RAPD technique in typing of 16 carbapenem-resistant P. aeruginosa strains isolated in 2007 from different patients of University HospitalNo. 1 of dr A. Jurasz Collegium Medicum of L. Rydygier in Bydgoszcz Nicolaus Copernicus University in Toruń. Study shows increasing frequency of isolation that type of strains when compared to 2006. Percentage of carbapenem-resistant isolates raised from 12,4% in 2006 to 22.9% in 2007. The majority of examined strains were obtained from patients of the Intensive Care Units (25.0%) and were isolated from bronchoalveolar lavage (25.0%), urine (25.0%) and wound swabs (18.8%) samples. Examined P. aeruginosa strains demonstrated resistance to doripenem (81.3%) and piperacillin (75.0%) and susceptibility to colistin (100.0%), amikacin (81.3%), netilmicin and norfloxacin (75.0% each). Using PCR-RAPD amplification with 208 and 272 primers, 14 and 16 DNA patterns were obtained, respectively. Usefulness of PCR-RAPD in carbapenem-resistant P. aeruginosa strains typing was proved in case of strains presenting similar and/or different antimicrobials susceptibility patterns.

  9. Genome relationships among Lotus species based on random amplified polymorphic DNA (RAPD).

    Science.gov (United States)

    Campos, L P; Raelson, J V; Grant, W F

    1994-06-01

    The ability of random amplified polymorphic DNA (RAPD) to distinguish among different taxa of Lotus was evaluated for several geographically dispersed accessions of four diploid Lotus species, L. tennis Waldst. et Kit, L. alpinus Schleich., L. japonicus (Regel) Larsen, and L. uliginosus Schkuhr and for the tetraploid L. corniculatus L., in order to ascertain whether RAPD data could offer additional evidence concerning the origin of the tetraploid L. corniculatus. Clear bands and several polymorphisms were obtained for 20 primers used for each species/accession. The evolutionary pathways among the species/accessions presented in a cladogram were expressed in terms of treelengths giving the most parsimonious reconstructions. Accessions within the same species grouped closely together. It is considered that L. uliginosus which is most distantly related to L. corniculatus, may be excluded as a direct progenitor of L. corniculatus, confirming previous results from isoenzyme studies. Lotus alpinus is grouped with accessions of L. corniculatus, which differs from previous studies. With this exception, these findings are in agreement with previous experimental studies in the L. corniculatus group. The value of the RAPD data to theories on the origin of L. corniculatus is discussed.

  10. Characterization of Pseudomonas syringae pathovars from different sweet cherry cultivars by RAPD analysis

    Directory of Open Access Journals (Sweden)

    Iličić Renata

    2016-01-01

    Full Text Available Pseudomonas syringae pvs., isolated from sweet cherry grown on different localities in Serbia, were genetically characterized using RAPD analysis. Four out of eleven tested primers (SPH1, DJP 17, DJ 15, and DJ 16 were selected on the basis of the differences between isolates within two pathovars - syringae and morsprunorum race 1. Cumulative RAPD analysis indicated heterogeneity within the population of both groups of tested isolates, revealing four different patterns in each group. RAPD analysis showed up to 24% differences among pv. syringae isolates, as well as 41% in comparison with the reference strain KFB0103 (pv. syringae, while differences of 15% among isolates pv. morsprunorum 1 race and 36% compared to the reference strain CFBP2119 (pv. morsprunorum 1 were observed. Isolates from locality Selenca exhibited three different genotypic patterns of pv. morsprunorum race 1 and one pattern of pv. syringae. Isolates of pv. morsprunorum collected in the same year from two plant organs (branches and leaves of the cv. Vanda yielded two different patterns. The pv. morsprunorum on cv. Kordia and pv. syringae on cv. Regina were detected at Mikicevo locality. The same patterns were observed for isolates of pv. syringae from Kanjiza and Selenca, as well as from Gornji Tavankut in two years of isolation. Differences were noted between isolates from the same pathovar originating from Ljutovo and Mikicevo, as well as with respect to all other isolates of same pathovar. [Projekat Ministarstva nauke Republike Srbije, br. III46007

  11. Detection of toxin genes and RAPD analysis of bacillus cereus isolates from different soil types

    Directory of Open Access Journals (Sweden)

    Savic Dejana

    2015-01-01

    Full Text Available The aim of this study was to detect genes for enterotoxins (hbla, entFM and bceT and for emetic toxin (cer, to determine antibiotic resistance, and to estimate intraspecies diversity in B. cereus isolates by RAPD analysis. B. cereus was identified in 12 out of 117 indigenous Bacillus spp. using the classical microbiological methods and PCR. All isolates were resistant to penicillin and ampicillin, two to tetracyclin and four to trimethoprim-sulphamethoxazole. Also, all isolates produced inducible penicillinases and β-lactamase. Toxin genes were detected with PCR. EntFM and cer genes were present in all isolates, hbla in all, but two, and bceT in none. RAPD analysis was performed with four different primers, two of them designed for this study. The intraspecies diversity revealed 10 different patterns at the 90% similarity level. Two separate clusters were formed regardless of a soil type or utilization. The detection of genes encoding toxins in all B. cereus isolates indicated these bacteria as potentially pathogenic and seriously for human health. Regardless of a soil type or utilization, the RAPD analysis showed high intraspecies heterogeneity in B. cereus isolates. To the best of our knowledge, this is the first study to analyse the presence of entero- and emetic toxin genes and genetic heterogeneity in B. cereus isolates from different soil types and different soil utilization in Serbia. [Projekat Ministarstva nauke Republike Srbije, br. TR37006

  12. Molecular differentiation of sheep and cattle isolates of Fasciola hepatica using RAPD-PCR

    Directory of Open Access Journals (Sweden)

    Khakpour, M.

    2012-11-01

    Full Text Available Understanding genetic structure and status of genetic variation of Fasciola hepatica isolates from different hosts, has important implications on epidemiology and effective control of fasciolosis. Random amplified polymorphic DNA (RAPD-PCR was used to study the genetic variation of F. hepatica in sheep and cattle. DNA was extracted from adult helminthes removed from livers of each infected animal in slaughterhouse at East-Azerbaijan province, North-West of Iran. DNA template amplified by the polymerase chain reaction, using three oligonucleotide decamers with arbitrary DNA sequences as primers. RAPD patterns showed the specific but different pattern DNA patterns for each primer. The intraspecific similarity coefficient within two isolates of F. hepatica was ranged between 69 to 100%. Present findings showed that the interspecific genetic distance was higher than intraspecific genetic distances (19-47% compares to 0-19%. Pair wise similarity matrices generated from each isolates-primer combination were totaled and the similarity coefficient between strains were calculated both manually (Nei and Li method and software analysis (Free-Tree-Freeware program. The inferred phylogenetic tree on the fingerprinting of these isolates clearly demonstrated the existence of population genetic diversity sub structuring within F. hepatica of sheep and cattle of Iran, raising interesting questions on the host specificity, epidemiology (e.g., zoonotic transmission and ecology of this fluke. RAPD-PCR is useful for both individual identification and epidemiological investigations in endemic regions.

  13. Genetic relatedness of soybean genotypes based on agromorphological traits and RAPD markers

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    Perić Vesna

    2014-01-01

    Full Text Available Modern agriculture, breeding procedures, as well as competition among breeding institutions contribute to further reduction of already narrowed diversity of soybean commercial varieties. The objective of the study was to characterize eighteen soybean cultivars from three different breeding programs for agro-morphological traits and to reveal genetic diversity using molecular markers. Morphological description was performed with 13 qualitative and 9 quantitative traits. The genetic relationships were estimated using 21 RAPD markers. PIC was calculated for RAPD data, while the diversity of qualitative traits was described by Shannon genetic diversity index. Cluster analysis based on qualitative morphological characters showed clear separation of genotypes on the basis of their plant growth type. PC analysis performed for quantitative traits divided genotypes according to their maturity group. Grouping pattern based on molecular marker data was in agreement with pedigree of cultivars. A great similarity was found, primarily between the varieties under the same institution, and then among all examined varieties. Comparison of three methods in the assessment of diversity indicated that morphological markers might provide useful information in breeding process and allow classification by pedigree to some extent, but RAPD markers were found to be superior in assessing differences among genetically very similar genotypes. [Projekat Ministarstva nauke Republike Srbije, br. TR-31068

  14. 异地板蓝根基因组DNA指纹图谱建立及RAPD-PCR反应体系优化%Establishment of Offsite Radix Isatidis Genomic DNA Fingerprint and Optimization of RAPD-PCR Reaction System

    Institute of Scientific and Technical Information of China (English)

    姜颖; 杨欣; 于英君

    2014-01-01

    Objective:RAPD method is used to study Radix Isatidis genomic DNA genetic difference and the true and false of the medicinal materials in different regions.Methods:Use RAPD technology for Radix Isatidis genomic DNA genetic diversity analyses at the molecular level from different regions, and optimize the Radix Isatidis RAPD-PCR reaction system.Results:Fingerprints showed in different regions Radix Isatidis resource had rich genetic diversity, and could distinguish between true and false, and the pros and cons of herbs.Conclusion:RAPD technique for the identification of the authenticity and the pros and cons of Chinese herbal medicines Radix Isatidis provides a practical significance for clinical safety and provide a theoretical basis for rational drug use.And Radix Isatidis best RAPD-PCR reaction system has been established.%目的:采用RAPD法研究不同地区板蓝根基因组DNA遗传差异性,药材的真假。方法:利用RAPD( random amplified polymorphic DNA)技术,在分子水平上对不同地区板蓝根基因组DNA进行了遗传差异性分析,同时优化板蓝根RAPD-PCR反应体系。结果:指纹图谱显示出不同地区板蓝根资源丰富的遗传多样性,可以辨别出药材的真假和优劣。结论:RAPD技术为鉴别中药材板蓝根真伪和优劣提供了现实意义,为临床安全和合理用药提供理论依据,并建立适合板蓝根RAPD-PCR的最佳反应体系。

  15. Application of the Random Amplified Polymorphic DNA (RAPD) Fingerprinting to Analyze Genetic Variation in Community Associated-Methicillin Resistant Staphylococcus Aureus (CA-MRSA) Isolates in Iran

    Science.gov (United States)

    Mobasherizadeh, Sina; Shojaei, Hasan; Havaei, Seyed Asghar; Mostafavizadeh, Kamyar; Davoodabadi, Fazollah; Khorvash, Farzin; Ataei, Behrooz; Daei-Naser, Abbas

    2016-01-01

    The aim of this study was to apply RAPD technique to analyze the genetic variability among the Iranian CA-MRSA isolates. The RAPD amplification was implemented on 25 strains isolated from the anterior nares of 410 healthy children using four randomly selected oligonucleotide primers from the stocks available in our laboratory, including the primers 1254, GE6, OLP6 and OLP13 from our stock. The amplified PCR products were detected on a 1.5% agarose gel and subjected to further analysis to establish the band profiles and genetic relationships using the Gel Compar® program. The Iranian CA-MRSA isolates produced distinct RAPD patterns which varied based on the primer used, however, the primer 1254 revealed highly polymorphic patterns consisting 5 discernable RAPD types (RT), “RT1” (12, 48%), “RT2” (8, 32%), “RT3” (3, 12%), and “RT4 and RT5”, (a single RAPD type each, 4%). Phylogenetic analysis based on RAPD profiles divided most of the CA-MRSA isolates into 2 distinct but related RAPD clusters, a small group and two single unrelated RAPD types. This study shows that the simple and cost-effective but rather difficult to optimize RAPD fingerprinting could be used to evaluate genetic and epidemiological relationships of CA-MRSA isolates on condition that the patterns are obtained from carefully optimized laboratory tests. PMID:27045409

  16. Identification, cloning and sequence analysis of a dwarf genome-specific RAPD marker in rubber tree [Hevea brasiliensis (Muell.) Arg].

    Science.gov (United States)

    Venkatachalam, P; Priya, P; Amma, C K Saraswathy; Thulaseedharan, A

    2004-11-01

    High-yielding dwarf clones of Hevea brasiliensis are tolerant to wind damage and therefore useful for high-density planting. The identification of molecular markers for the dwarf character is very important for isolating true-to-type high-yielding dwarf hybrid lines in the early stage of plant breeding programs. We have identified a dwarf genome-specific random amplified polymorphic DNA (RAPD) marker in rubber tree. A total of 115 random oligonucleotide 10-mer primers were used to amplify genomic DNA by PCR, of which 19 primers produced clear and detectable bands. The primer OPB-12 generated a 1.4-kb DNA marker from both natural and controlled F(1) hybrid progenies (dwarf stature) derived from a cross between a dwarf parent and a normal cultivated clone as well as from the dwarf parent; it was absent in other parent (RRII 118). To validate this DNA marker, we analyzed 22 F(1) hybrids (13 with a dwarf stature and nine with a normal stature); the dwarf genome-specific 1.4-kb RAPD marker was present in all dwarf-stature hybrids and absent in all normal-stature hybrids. This DNA marker was cloned and characterized. DNA marker locus specificity was further confirmed by Southern blot hybridization. Our results indicate that Southern blot hybridization of RAPD using probes made from cloned DNA fragments allows a more accurate analysis of the RAPD pattern based on the presence/absence of specific DNA markers than dye-stained gels or Southern blot analysis of RAPD blots using probes made from purified PCR products. Detection of RAPD markers in the hybrid progenies indicates that RAPD is a powerful tool for identifying inherited genome segments following different hybridization methods in perennial tree crops.

  17. A comparative phylogenetic analysis of medicinal plant Tribulus terrestris in Northwest India revealed by RAPD and ISSR markers

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    ASHWANI KUMAR

    2012-07-01

    Full Text Available Kumar A, Verma N. 2012. A comparative phylogenetic analysis of medicinal plant Tribulus terrestris in Northwest India revealed by RAPD and ISSR markers. Biodiversitas 13: 107-113. Several DNA marker systems and associated techniques are available today for fingerprinting of plant varieties. A total of 5 RAPD and 8 ISSR primers were used. Amplification of genomic DNA of the 6 genotypes, using RAPD analysis, yielded 164 fragments that could be scored, of which 47 were polymorphic, with an average of 9.4 polymorphic fragments per primer. Number of amplified fragments with random primers ranged from 6 (AKR-1 to 10 (AKR-4 and varied in size from 200 bp to 2,500 bp. Percentage polymorphism ranged from 16% (AKR-4 to a maximum of 41% (AKR-4, with an average of 29.6%. The 8 ISSR primers used in the study produced 327 bands across 6 genotypes, of which 114 were polymorphic. The number of amplified bands varied from 7 (ISSR 7 to 12 (ISSR 1&3, with a size range of 250-2,800 bp. The average numbers of bands per primer and polymorphic bands per primer were 40.87 and 14.25, respectively. Percentage polymorphism ranged from 24% (ISSR 4 to 53.84% (ISSR 2, with an average percentage polymorphism of 35.59% across all the genotypes. The 3′-anchored primers based on poly (AC and poly (AT motifs produced high average polymorphisms of 53.84% and 40.81%, respectively. ISSR markers were more efficient than the RAPD assay, as they detected 35.59% polymorphic DNA markers in Tribulus terrestris as compared to 29.6% for RAPD markers. Clustering of genotypes within groups was not similar when RAPD and ISSR derived dendrogram were compared, whereas the pattern of clustering of the genotypes remained more or less the same in ISSR and combined data of RAPD and ISSR.

  18. Efficacy of liposomal amphotericin B (AmBisome) in the treatment of persistent post-kala-azar dermal leishmaniasis (PKDL).

    Science.gov (United States)

    Musa, A M; Khalil, E A G; Mahgoub, F A; Hamad, S; Elkadaru, A M Y; El Hassan, A M

    2005-09-01

    A dermatosis commonly known as post-kala-azar dermal leishmaniasis (PKDL) may develop following the treatment of human visceral leishmaniasis (VL). In about 15% of PKDL cases the disfiguring lesions persist, sometimes for many years. Such persistent lesions currently require daily injections of sodium stibogluconate (SSG) for 2-4 months and even then treatment may not be successful. Alternative, quicker and cheaper treatment options that cause less toxicity are being explored. Immuno-chemotherapeutic regimens (based on leishmaniasis candidate vaccines/BCG with SSG) are still experimental but treatment with liposomal amphotericin B (AmBisome) has already been found effective, albeit in a small number of patients. AmBisome is considered less nephrotoxic than non-liposomal amphotericin B because it specifically targets the macrophages in which the Leishmania parasites develop. The aim of the present study was to evaluate further the usefulness of AmBisome in the treatment of persistent PKDL, in Sudan. The 12 subjects, all of whom gave their informed consent, had each had PKDL lesions for >6 months and shown no improvement after repeated injections of SSG. During the study period, they were hospitalized and regularly screened, haematologically and biochemically, for adverse effects. The AmBisome, given intravenously at 2.5 mg/kg.day for 20 days, completely cleared the skin rash of 10 (83%) of the patients and caused no detectable adverse effects. In the 10 patients who responded well to the treatment, the papular lesions regressed and became flat while the hypopigmented lesions darkened (continuing to do so even after the last AmBisome injections). Treatment outcome appeared to be unaffected by the age or gender of the patient (P = 0.7 for each) but the time taken for the PKDL lesions to heal was correlated with the age of the lesions (P = 0.009). The macular lesions healed more slowly than the papular (P = 0.02). In conclusion, Ambisome appears suitable for the

  19. Gliricidia sepium al establecimiento

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    J. L. Valle

    2004-01-01

    Full Text Available Se asoció Cenchrus ciliaris (Cc y Gliricidia sepium (Gs para evaluar la producción de biomasa y la composición química de los forrajes en la fase de establecimiento, en el estado de Morelos, México, en condiciones de trópico seco. Se empleó un análisis de varianza con diseño en bloques al azar, en donde T1 fue pasto solo, T2 Cc más Gs con 5,000 plantas ha-1 y el T3 Cs más Gs con 14,285 plantas ha-1. Se midió la producción de forraje individual y asociado con materia seca (t MS/Ha, la altura (A en cm, la proteína cruda (PC%, las fracciones de fibra (FDN% y FDA% y la digestibilidad in vitro de la materia seca (DIVMS%. El T3 tuvo una mejor producción de biomasa (P0.05. La asociación de 14,285 plantas ha-1 de G. sepium con pasto Cenchrus ciliaris mejoró la producción de biomasa y la disponibilidad total de nutrientes por superficie cultivada al establecimiento

  20. Ludomania: avaliação transcultural do jogo de azar por dinheiro e seu tratamento Ludomania: cross-cultural examinations of gambling and its treatment

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    Jeremiah Weinstock

    2008-05-01

    Full Text Available O jogo patológico é um transtorno do controle do impulso que está ganhando mais e mais atenção. Este artigo revisa os critérios diagnósticos e os instrumentos de rastreamento para o jogo patológico, bem como os índices de prevalência desse transtorno ao redor do mundo, com ênfase na situação do jogo de azar no Brasil. Os tratamentos para o jogo patológico são também descritos, incluindo tanto as abordagens psicossociais como as farmacológicas. O jogo patológico é altamente comórbido com outros transtornos psiquiátricos, incluindo abuso de substâncias e depressão e poucos jogadores patológicos buscam tratamento para seus problemas com o jogo de azar. Portanto, recomenda-se a procura direta de problemas com o jogo de azar. É necessário melhorar a educação sobre o transtorno, tanto sob a perspectiva do cuidador como da sociedade, a fim de reduzir as conse-qüências pessoais e sociais desse transtorno.Pathological gambling is a disorder of impulse control that is gaining more and more attention. This paper reviews diagnostic criteria and screening instruments for pathological gambling, as well as the prevalence rates of this disorder worldwide, with an emphasis on gambling in Brazil. Treatments for pathological gambling are also described, including both psychosocial and pharmacological approaches. Pathological gambling is highly comorbid with other psychiatric disorders, including substance abuse and depression, and few pathological gamblers seek treatment for their gambling problems. Therefore, direct screening for gambling problems is recommended. Increasing education about the disorder, from both the provider and societal perspective, is necessary to reduce the personal and societal consequences of this disorder.

  1. Adaptación de la estructura organizacional en la empresa Apuestas Azar S.A. dentro de su proceso evolutivo actual

    OpenAIRE

    2011-01-01

    El diseño de una estructura que funcione en una organización presupone que si un ejecutivo, puede colocar las adecuadas casillas del organigrama, establecer entre ellas la apropiada jerarquía y poner en su frente buenos hombres que las dirijan, la empresa será un éxito. Con esa finalidad, la mayoría de empresas se embarcan en importantes reorganizaciones. Las consideraciones presentadas para la adaptación de la estructura organizacional de la empresa Apuestas Azar, tiene en cuenta la empr...

  2. Analysis of Genetic Diversity in Cultivated and Wild Tomato Varieties in Chinese Market by RAPD and SSR

    Institute of Scientific and Technical Information of China (English)

    MENG Fan-juan; XU Xiang-yang; HUANG Feng-lan; LI Jing-fu

    2010-01-01

    RAPD and SSR were applied to assess genetic diversity in 61 tomato varieties from different species (Solanum lycopersicum L.,hirsutum.Humb L.,pimpinellifolium Miller L.,chilense Dun.L.,chmielenskii L.,peruvianum Miller L.,parvuflorum Miller L.).2062 and 869 clear fragments were amplified by RAPD and SSR,respectively.On the other hand,more polymorphic products were found by SSR as compared to RAPD,i.e.,100 and 43.84%,respectively.In addition,a higher value of the average similarity coefficient and lower PIC value were reflected in RAPD (0.79,0.407) compared to SSR (0.56,0.687).It can be inferred that SSR was a higher effective marker than RAPD to assess genetic diversity in tomato accessions.Similarly,the genetic base of tomato varieties in Chinese market was narrow.It is suggested that wild tomato varieties should be used to enrich the genetic base of the cultivated tomato varieties.

  3. Clones identification of Sequoia sempervirens (D.Don) Endl.in Chile by using PCR-RAPDs technique

    Institute of Scientific and Technical Information of China (English)

    Manuel TORAL IBA(N)EZ; Margarita CARU; Miguel A.HERRERA; Luis GONZALEZ; Luis M.MARTIN; Jorge MIRANDA; Rafael M.NAVARRO-CERRILLO

    2009-01-01

    A protocol of polymerase chain reaction-random amplified polymorphic DNAs (PCR-RAPDs) was established to analyse the gene diversity and genotype identification for clones of Sequoia sempervirens (D.Don) Endl.in Chile.Ten (out of 34) clones from introduction trial located in Voipir-Viilarrica,Chile,were studied.The PCR-RAPDs technique and a modified hexadecyltrimethylammonium bromide (CTAB) protocol were used for genomic DNA extraction.The PCR tests were carried out employing 10-mer random primers.The amplification products were detected by electrophoresis in agarose gels.Forty nine polymorphic bands were obtained with the selected primers (BG04,BF07,BF 12,BF13,and BF 14) and were ordered according to their molecular size.The genetic similarity between samples was calculated by the Jaccard index and a dendrogram was constructed using a cluster analysis of unweighted pair group method using arithmetic averages (UPGMA).Of the primers tested,5 (out of 60) RAPD primers were selected for their reproducibility and high polymorphism.A total of 49 polymorphic RAPD bands were detected out of 252 bands.The genetic similarity analysis demonstrates an extensive genetic variability between the tested clones and the dendrogram depicts the genetic relationships among the clones,suggesting a geographic relationship.The results indicate that the RAPD markers permitted the identification of the assayed clones,although they are derived from the same geographic origin.

  4. Análisis de la diversidad genética de 21 aislamientos del hongo Moniliophthora roreri basado en marcadores RAPD

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    Boris Gutarra Castillo

    2013-12-01

    Full Text Available Objetivos: Estudiar la diversidad genética de 21 aislamientos del hongo que afecta al cultivo del cacao, Moniliophthora roreri, en tres zonas cacaoteras del Perú (Tocache, Mariscal Cáceres y Leoncio Prado. Métodos: Se utilizó 14 iniciadores RAPD (random amplified polymorphic DNA polimórficos y una pareja de oligonucleótidos, los que fueron empleados bajo condiciones de amplificación estandarizadas. Con los datos obtenidos se construyó un dendograma utilizando el coeficiente de Jaccard y el algoritmo UPGMA (Unweighted Pair-Group Method using Arithmetic Average. La estructura genética fue estimada en función del análisis molecular de variancia (AMOVA y la diversidad mediante los índices de Shannon y Nei. Resultados: Fueron conseguidas 59 bandas RAPD con un 73% de polimorfismo. El dendograma obtenido a un índice de similitud de 0,70, claramente dividió los individuos en tres grupos. El análisis de la diversidad genética mostró altos valores en las zonas estudiadas de acuerdo con el índice de Shannon (0,3936 y de Nei (0,2622, con mayor riqueza en Leoncio Prado. Estas zonas presentan alta variabilidad, y según el AMOVA realizado: 88% entre accesiones por zona y solo 12% entre zonas. Conclusiones: Existe más de un grupo genético de Moniliophthora roreri en la Amazonía del Perú. Estos grupos, provenientes del Ecuador, pudieron haber ingresado por el intercambio de semillas y/o de forma natural por medio de los ríos en común y estarían originando nuevos grupos genéticos locales.

  5. Datziinae as a new subfamily name for the unavailable name Protopsychodinae Stebner et al., 2015, (Diptera: Psychodidae

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    Frauke Stebner

    2015-11-01

    Full Text Available In a recent paper a new subfamily of Psychodidae was inadequately named Protopsychodinae. This nomenclatural act cannot be considered as a valid name under ICZN regulations because the subfamily name is not based on the type genus Datzia Stebner et al., 2015, and furthermore the fossil genus Protopsychoda Azar et al., 1999 was originally described under the subfamily Psychodinae. Therefore, the new family-group name Datziinae is herein proposed.

  6. Molecular heterogeneity of Malassezia pachydermatis through RAPD-PCR = Heterogeneidade molecular da Malassezia pachydermatis através de RAPD-PCR

    Directory of Open Access Journals (Sweden)

    Patrícia da Silva Nascente

    2010-04-01

    Full Text Available Several methodologies in molecular biology have been used in theinvestigation of Malassezia pachydermatis and its differentiation into subtypes. Recent molecular research of this species includes the use of samples isolated from canine otitis externa and dermatitis, as well as from healthy animals, having in view an epidemiologicalstudy of the yeast. The aim of this study was to identify molecular differences in M. pachydermatis samples isolated from dogs with otitis externa. The M. pachydermatis strains were analyzed by means of the Random Amplification Primer DNA - Polimerase Chain Reaction (RAPD–PCR for molecular heterogeneity research. DNA extraction was carried out with phenol-chloroform and the RAPD technique using the AGAATCCGCC primer. A variation was observed in the number and arrangement of the bands among the 49 studied isolates, grouped into nine patterns. Isolate groupings were not found to be related to animal breed, age or sex. It was concluded that M. pachydermatis has differences in its molecular profile, as shown by the molecular technique (RAPD – PCR, which allows isolates to be classified into nine subtypes.Várias metodologias em biologia molecular têm sido aplicadas para estudar a M. pachydermatis diferenciando-a em subgrupos. Recentemente utiliza-se a investigação molecular desta espécie isolada de otite externa e dermatite, e também de isolados da mesma de animais hígidos, para um estudo epidemiológico da levedura. O objetivo deste trabalho foi identificar diferenças moleculares entre isolados de M. pachydermatis obtidos de casos de otite externa canina. Para isto, amostras da levedura provenientes de cães com esta enfermidade foram estudadas através da técnica de Polimorfismo de DNA Amplificado aoAcaso - Reação da Polimerase em Cadeia (RAPD–PCR para pesquisa de heterogeneidade molecular. A extração de DNA foi realizada no processo fenol-cloroformio e a técnica de RAPD foi estudada com o primer

  7. Genetic distances in soybean based on RAPD markers Distâncias genéticas em soja com base em marcadores RAPD

    Directory of Open Access Journals (Sweden)

    RONAN XAVIER CORRÊA

    1999-01-01

    Full Text Available Four methods were applied to determine pairwise genetic distances among five soybean genotypes which are potential genitors for a mapping population. Additionally, individual plants from the most divergent pair of genotypes were evaluated by the RAPD technique to determine their degree of homozygosity. Genetic distances based on RAPD data were calculated by the modified Rogers' distance, and also by the following arithmetical complements of similarity: simple match, Nei and Li, and Gower. These genetic distances were similar, presenting a correlation coefficient ranging from 0.99 to 1.00. In all four methods lines UFV 91-717 and Ichigowase were the most divergent ones (4.53 to 21.43%. DNA samples from five plants from each of the two most divergent genotypes were amplified with 28 different primers. Among the amplified products, only five were polymorphic in each group (2.10%, demonstrating their high intragroup degree of homozygosity. These homozygosity were maintained when DNA samples from 12 plants from each of the two most divergent genotypes were amplified. These parameters were extremely useful for the confirmation of the chosen pair of genitors to generate a mapping population.Aplicaram-se quatro métodos para determinar as distâncias genéticas entre cinco cultivares de soja, que são genitores potenciais para uma população de mapeamento genético. Adicionalmente, o grau de homozigose do par de genótipos mais divergente foi avaliado por meio da técnica de RAPD. Calcularam-se as distâncias genéticas fundadas em dados obtidos pela técnica de RAPD pela distância modificada de Rogers e pelos seguintes complementos aritméticos de similaridade: distância simples; Nei e Li, e Gower. As distâncias genéticas foram similares, apresentando valores de coeficiente de correlação de 0,99 a 1,00. Nos quatro métodos, as linhagens UFV 91-717 e Ichigowase foram as mais divergentes (4,53 to 21,43%. Amostras de DNA de cinco plantas de cada

  8. High-resolution melt-curve analysis of random amplified polymorphic DNA (RAPD-HRM) for the characterisation of pathogenic leptospires

    DEFF Research Database (Denmark)

    Tulsiani, Suhella; Craig, S B; Graham, G C;

    2010-01-01

    High-resolution melt-curve analysis of random amplified polymorphic DNA (RAPD-HRM) is a novel technology that has emerged as a possible method to characterise leptospires to serovar level. RAPD-HRM has recently been used to measure intra-serovar convergence between strains of the same serovar...

  9. Prevalence and control of kala-azar in China%我国黑热病的流行概况和防治现状

    Institute of Scientific and Technical Information of China (English)

    李玉凤; 仲维霞; 赵桂华; 王洪法

    2011-01-01

    Kala-azar (commonly known as visceral leishmaniasis), is serious harm to human health as a parasitic disease. It can be classified in to three types of diseases:wild visceral leishmaniasis, caninotic visceral leishmaniasis and an-throponotic visceral leishmaniasis. At present, anthroponotic visceral leishmaniasis has been controlled in most areas except Xinjiang. But caninotic visceral leishmaniasis and anthroponotic visceral leishmaniasis also appears at times in their endemic areas. The paper discusses and reviews the disease's classification, the current epidemic situation and the control measures of kala-azar in our country.%黑热病又称内脏利什曼病,是严重危害人类身体健康的寄生虫病.按传染源特点可分为野生动物源型、犬源型和人源型3种类型.目前,人源型黑热病除新疆流行区外,在其他流行区已得到控制.而犬源型和野生动物源型黑热病则在其流行区不断出现,有死灰复燃之势.本文对我国黑热病的分型、流行概况和防治现状进行了综述.

  10. Appraisal of Phlebotomus argentipes habitat suitability using a remotely sensed index in the kala-azar endemic focus of Bihar, India

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    Shreekant Kesari

    2013-04-01

    Full Text Available Visceral leishmaniasis, or kala-azar, is recognised as a serious emerging public health problem in India. In this study, environmental parameters, such as land surface temperature (LST and renormalised difference vegetation indices (RDVI, were used to delineate the association between environmental variables and Phlebotomus argentipes abundance in a representative endemic region of Bihar, India. The adult P. argentipes were collected between September 2009-February 2010 using the hand-held aspirator technique. The distribution of P. argentipes was analysed with the LST and RDVI of the peak and lean seasons. The association between environmental covariates and P. argentipes density was analysed a multivariate linear regression model. The sandfly density at its maximum in September, whereas the minimum density was recorded in January. The regression model indicated that the season, minimum LST, mean LST and mean RDVI were the best environmental covariates for the P. argentipes distribution. The final model indicated that nearly 74% of the variance of sandfly density could be explained by these environmental covariates. This approach might be useful for mapping and predicting the distribution of P. argentipes, which may help the health agencies that are involved in the kala-azar control programme focus on high-risk areas.

  11. Síndrome de Felty e Calazar: um desafio para o reumatologista Felty's syndrome and Kala-azar: a challenge for the rheumatologist

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    Rafaela Bicalho Viana

    2010-12-01

    Full Text Available Os autores relatam o caso de uma paciente com artrite reumatoide que evoluiu com grave neutropenia e esplenomegalia, sendo firmado o diagnóstico de Síndrome de Felty, que posteriormente desenvolveu Calazar. Ambas têm apresentação clínica e laboratorial semelhantes, tornando o diagnóstico diferencial difícil. O relato deste caso objetiva chamar a atenção para o reconhecimento da infecção por leishmaniose visceral em pacientes portadores de doenças reumáticas, assim como a possibilidade de um paciente com Calazar mimetizar um quadro de doença reumática sistêmicaCase report of a patient with rheumatoid arthritis who developed severe neutropenia, splenomegaly and was diagnosed with Felty's syndrome. The patient later developed Kala-azar. Both diseases have similar clinical and laboratory presentation, making the differential diagnosis difficult. The present case report aims at drawing attention to the identification of visceral Leishmaniasis infection in patients with rheumatic diseases, as well as possibility of a patient with Kala-azar mimicking a set of symptoms of systemic rheumatic disease

  12. Intra-specific relationships among Tibetan Eared-pheasants based on randomly amplified polymorphic DNA(RAPD) analysis

    Institute of Scientific and Technical Information of China (English)

    DING Wei; ZHANG Zhengwang; CHANG Jiang; ZHANG Er; WU Xiushan; ZHANG Jinguo

    2006-01-01

    The Tibetan Eared-pheasant Crossoptilon harmani is a rare species native to China.A captive population has been established in the Beijing Zoo since 1999.In order to determine the kinship of the offsprings in 2001,randomly amplified polymorphic DNA (RAPD) was used to examine the parenthood of seven Tibetan Eared-pheasants in the Beijing Zoo.To amplify the genomic DNA of each individual,53 arbitrary primers were selected.The results of amplifica tions showed that 14 primers had clear and distinct RAPD patterns.Totally,226 amplified fragments were generated by RAPD in this study.Cluster analysis of the seven Tibetan Eared-pheasants indicated that all the four young birds had the same father (No.5 male).This study provides a practical method to determine the relationship of offsprings whose parents are unknown in birds.

  13. Genetic variation detected by RAPD markers in natural populations of babassu palm (Attalea speciosa Mart.).

    Science.gov (United States)

    Santos, M F; Damasceno-Silva, K J; Carvalhaes, M A; Lima, P S C

    2015-06-10

    The purpose of this study was to analyze the effects of management on the genetic structure of natural populations of Attalea speciosa in the State of Piauí, Brazil, using random-amplified polymorphic DNA (RAPD) markers. Three babassu populations under different management systems were selected. Polymerase chain reactions were performed for 20 RAPD primers. A total of 146 bands were generated, 141 of which were polymorphic (96.58%), with a variation of 4 and 12 loci and an average of 7 bands per primer. A dendrogram revealed a clear separation between the three populations (0.57). Data reliability and node consistency were verified by bootstrap values and the cophenetic correlation coefficient (88.15%). Coefficients of similarity between pairs of genotypes ranged from 0.26 to 0.86, with a mean of 0.57. Nei's genetic diversity index (HE) value of the population sampled in Teresina was 0.212, of Esperantina it was 0.195, and of José de Freitas it was 0.207. After the HE was decomposed, the complete diversity was found to be 0.3213. Genetic differentiation between populations was 0.362, and the estimation of gene flow between populations was low (0.879). Analysis of molecular variance revealed that 59.52% of the variation was contained within populations, and 40.48% was between populations. RAPD markers were effective for genetic diversity analysis within and between natural babassu populations, and exhibited a high level of polymorphism. Genetic diversity was the highest within populations; variability was lower in the managed populations than in the undisturbed populations.

  14. AFLP and RAPD Analysis of the Boer and Indigenous Breeds of the Goat in Jiangsu

    Institute of Scientific and Technical Information of China (English)

    CAO Shao-xian; YANG Li-guo; JIANG Xun-ping; LIU Hong-lin; LU Wei-zhong; XIANG Yang-hai

    2003-01-01

    Blood and tissue samples were collected from 105 goats including 60 Boer goats (30 for eachsex), 30 Xuhuai goats (15 for each sex) and 15 Haimen goats (7 stud and 8 does). DNA was extracted andDNA pools were constructed on the basis of goat breeds. In 36 selective primer combinations, 29 combinationsamplified totally 3 253 markers including 92 polymorphic markers by amplified fragment length polymor-phism (AFLP). On average, 3.17 polymorphic markers were amplified per combination, with a polymorphicfrequency of 2.8%. The primer combinations amplifying more polymorphic markers (showed in brackets)were involved in E00+ACG/M00+CAA (13), E00+ACG/M00+CAG (10), E00+AAC/M00+CAC (8)and E00+AAC/M00+ACT (7). A total of 183 markers including 60 polymorphic markers were amplified byRAPD from the pooled DNA of three breeds using 22 primers with strong polymorphism and high reproduc-ibility selected from 93 RAPD primers. On average, 2.73 polymorphic markers were amplified per primer,with a polymorphic frequency of 32.8%. The results of AFLP and RAPD coincidently suggested that the ge-netic distance is the closest between Xuhuai and Haimen goat, next between Xuhuai and Boer goat, and the far-thest between Haimen and Boer goat. According to the UPGMA method, Haimen and Xuhuai goats can begathered together as a cluster, then Boer goat. Both methods can be used to implicate the genetic difference ofthese three breeds, in particular AFLP has more polymorphic markers.

  15. Selection of parent trees for Rubber (Hevea brasiliensis breeding based on RAPD analysis

    Directory of Open Access Journals (Sweden)

    KUSWANHADI

    2011-11-01

    Full Text Available Oktavia F, Lasminingsih M, Kuswanhadi. 2011. Selection of parent trees for Rubber (Hevea brasiliensis breeding based on RAPD analysis. Nusantara Bioscience 3: 124-129. The parent trees’ clones usually originate from the previous generation having the potential of high production with better agronomical characters. Although, phenotype characters can determine the genetic variability among accessions, they are highly sensitive to environmental factors, so it is often difficult to identify the difference between closely related clones. The genetic variability or phylogenetic relationships among rubber clones can be analysis using RAPD method, and based on the result, the parent trees can be selected. This research was aimed to analyze the genetic distance among rubber clones using RAPD method. Analysis was conducted on 45 rubber clones with 12 random primers. Pair-wise comparisons of unique and shared polymorphic amplification products were used to generate similarity coefficients. These coefficients were employed to construct a dendogram by using an Unweighted Pair-Group Method with Arithmetical Averages (UPGMA. The amplification of genomic DNA from 45 clones yielded 2408 DNA fragments, ranging in size from 250 bp to 3000 bp. The range of genetic similarity matrix was very wide (59.18%-94.23%. It indicated that most of the clones have a low level of polymorphism. The lowest genetic similarity (59,18% was found between RRIC 110 and AVROS 352 clones, while the highest (94.23% was between IRR 41 and IRR 42 clones. Cluster analysis showed that 45 clones of rubber were divided into two groups, the biggest group consisted of 30 clones, while the other one consisted of 15 clones with a genetic similarity value of 0,73.

  16. Genetic Diversity Assessment of Acid Lime (Citrus Aurantifolia Swingle Landraces of Eastern Nepal Using RAPD Markers

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    NN Munankarmi

    2014-09-01

    Full Text Available Acid lime (Citrus aurantifolia Swingle is an important commercial fruit crop, cultivated from terai to high hill landscapes of Nepal. However, production and productivity is very low due to various reasons including infestations by various diseases and pests, lack of diseases and pests resistant and high yielding varieties. In this context, determination of genetic variation at molecular level is fundamental to citrus breeders for the development of elite cultivars with desirable traits. In the present study, Random Amplified Polymorphic DNA (RAPD marker technique has been employed to assess genetic diversity in 60 acid lime landraces representing different agro-ecological zones of eastern Nepal. Nine selected arbitrary primers generated 79 RAPD fragments of which 75 were polymorphic (94.94%. Phenogram was constructed by NTSYSPC ver. 2.21i using UPGMA cluster analysis based on Jaccard’s similarity coefficient to deduce overall genetic diversity and relationships of the acidlime genotypes under study. Sixty acid lime landraces formed seven clusters and similarity value ranged from 38% to 98% with an average of 72%. Genetic variation at different agro-ecological zones was assessed using Popgene ver. 1.32 and found 47% to 69.6% polymorphism. Shannon’s index and Nei’s gene diversity showed highest level of acid lime diversity in Terai zone (PPB, 69.62%; H, 0.213; I, 0.325 followed by mid-hill zone (PPB, 67.09%; H, 0.208; I, 0.317. The results obtained will be useful to citrus breeders for elite cultivar development. The RAPD-PCR technique is found to be the rapid and effective tool for genetic diversity assessment in acid lime landraces of Nepal.

  17. A phase III trial of efficacy of the FML-vaccine against canine kala-azar in an endemic area of Brazil (São Gonçalo do Amaranto, RN).

    Science.gov (United States)

    da Silva, V O; Borja-Cabrera, G P; Correia Pontes, N N; de Souza, E P; Luz, K G; Palatnik, M; Palatnik de Sousa, C B

    2000-12-08

    Protection against canine kala-azar was investigated in naturally exposed dogs of an endemic area, vaccinated with the fucose mannose ligand (FML)-vaccine of Leishmania donovani. A total of 97% of vaccinees were seropositive to FML and 100% showed intradermal reaction to L. donovani lysate, 7 months after vaccination. The absorbency values and size of intradermal reaction were both significantly higher in vaccinees than in controls (ANOVA, P<0.0001). After 2 years, 92% (chi(2)=6.996; P<0.0025) protection was achieved: only 8% of vaccinees showed mild signs of kala-azar with no deaths while 33% of controls developed clinical or fatal disease. The FML-vaccine induced a significant, long-lasting and strong protective effect against canine kala-azar in the field.

  18. Unraveling the efficiency of RAPD and SSR markers in diversity analysis and population structure estimation in common bean.

    Science.gov (United States)

    Zargar, Sajad Majeed; Farhat, Sufia; Mahajan, Reetika; Bhakhri, Ayushi; Sharma, Arjun

    2016-01-01

    Increase in food production viz-a-viz quality of food is important to feed the growing human population to attain food as well as nutritional security. The availability of diverse germplasm of any crop is an important genetic resource to mine the genes that may assist in attaining food as well as nutritional security. Here we used 15 RAPD and 23 SSR markers to elucidate diversity among 51 common bean genotypes mostly landraces collected from the Himalayan region of Jammu and Kashmir, India. We observed that both the markers are highly polymorphic. The discriminatory power of these markers was determined using various parameters like; percent polymorphism, PIC, resolving power and marker index. 15 RAPDs produced 171 polymorphic bands, while 23 SSRs produced 268 polymorphic bands. SSRs showed a higher PIC value (0.300) compared to RAPDs (0.243). Further the resolving power of SSRs was 5.241 compared to 3.86 for RAPDs. However, RAPDs showed a higher marker index (2.69) compared to SSRs (1.279) that may be attributed to their higher multiplex ratio. The dendrograms generated with hierarchical UPGMA cluster analysis grouped genotypes into two main clusters with various degrees of sub clustering within the cluster. Here we observed that both the marker systems showed comparable accuracy in grouping genotypes of common bean according to their area of cultivation. The model based STRUCTURE analysis using 15 RAPD and 23 SSR markers identified a population with 3 sub-populations which corresponds to distance based groupings. High level of genetic diversity was observed within the population. These findings have further implications in common bean breeding as well as conservation programs.

  19. Identification of ISSR and RAPD markers linked to yield traits in bread wheat under normal and drought conditions

    Directory of Open Access Journals (Sweden)

    A.G.A. Khaled

    2015-12-01

    Full Text Available Genetic variability and identification of some molecular markers were studied in twenty promising lines of wheat using agronomic traits, ISSR (inter simple sequences repeats and RAPD (random amplified polymorphic DNA markers. Significant variation was evidenced in all agronomic traits. The lines proved to be superior to the check cultivar Sahel1 in yield and its component traits. Lines L2, L7 and L8 were the best in most yield component traits in both seasons. Moreover, Lines L2, L4, L5, L7 and L8 showed drought tolerance by which they displayed high performance in agronomic traits as well as a low drought susceptibility index. The percentage of polymorphism was 39.3% and 53.2% for ISSRs and RAPDs, respectively. UBC-881 belonged to penta-nucleotide repeat sequences (GGGTG that produced the highest level of polymorphism, while UBC-846 belonged to di-nucleotide repeat sequences (CA that produced the lowest level of polymorphism. Genetic similarities among wheat lines based on ISSR and RAPD markers ranged from 0.81 to 1.00 and from 0.86 to 0.98, respectively. There was a low average of PIC (polymorphism information content values which were 0.10 (ISSR and 0.15 (RAPD. The RAPD technique exhibited a higher marker index (MI = 0.69 compared to ISSR (MI = 0.43. There was insignificant correlation between ISSR and RAPD data (0.168, p > 0.05. There were two markers (UBC-881450bp and OPF-10540bp, on each of which two traits regressed significantly. The associated markers each explained a maximum regression of 18.92–34.95% of the total available variation for individual associated traits.

  20. Analysis on Genotypic Differentiation of Phytophthora infestans by Using Random Amplified Polymorphic DNA (RAPD)

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    A total of 104 isolates including two Korean isolates and three Japanese isolates of Phytophthora infestans collected from Heilongjiang and Jilin Provinces from 2006 to 2008 were used to determine their mating types,metalaxyl resistance,and RAPD genotypes.All the isolates of P.infestans collected from Heilongjiang and Jilin Provinces belonged to A 1 mating type,and no A 2 mating type was detected.Frequencies of metalaxyl resistant isolates were 94.4%,47.8% and 75.0% in 2006,2007 and 2008,respectively.Accord...

  1. Genetic linkage map of Brassica campestris L. Using AFLP and RAPD markers

    Institute of Scientific and Technical Information of China (English)

    卢钢; 曹家树; 陈杭

    2002-01-01

    A genetic linkage map comprised of 131 loci was constructed with an F2 population derived from an inter-subspecific cross between Brassica 'qisihai'. The genetic map included 93 RAPD loci, 36 AFLP loci and 2 morphological loci organized into 10 main linkage groups (LGs) and 2 small groups, covering 1810.9cM with average distance between adjacent markers being approximately 13.8cM. The map is suitable for identification of molecular markers linked to important agronomic traits, QTL analysis, and even for marker-assisted selection in breeding programs of Chinese cabbage and turnip.

  2. Identification of species of leishmania isolated from patients with cutaneous leishmaniasis in Kermanshah; using RAPD-PCR technique

    Directory of Open Access Journals (Sweden)

    Yazdan Hamzavi

    2010-09-01

    Full Text Available Annually many numbers of pationts with Cutaneous Leishmaniasis (CL have been reported in Kermanshah province- IRAN. The study aimed to identify species of Leishmania isolated from patients with CT in Kermanshah. Seven isolates of Leishmania obtained from patients with CL, without any travelling to other provinces, were cultured in NNN medium. After mass production of leptomonads in RPMI 1640 medium DNA was purified and the species were diagnosed using RAPD-PCR technique. The study of electrophoretic fingerprints of the product of RAPD-PCR in seven isolates showed that Leshmania major was the causative agent of CL patients in Kermanshah province. More studies in this field recommended.

  3. Estudio comparativo, cruzado, doble ciego, al azar para determinar la bioequivalencia entre dos formulaciones de valsartán en tabletas y cápsulas

    Directory of Open Access Journals (Sweden)

    Milena Pérez

    2006-06-01

    Full Text Available Introducción: Los estudios de bioequivalencia o equivalencia comparada se realizan para demostrar que el producto en estudio, conocido como producto genérico, tiene la misma biodisponibilidad del producto de referencia, también conocido como producto innovador o de marca. Si los dos productos son bioequivalentes, se espera que tengan las mismas características de seguridad y eficacia. La bioequivalencia es establecida por la estimación estadística de diferencias significativas o no en los parámetros farmacocinéticos de área bajo la curva (ABC y concentración máxima (Cmáx. En este caso, se evaluará y se comparará la biodisponibilidad de valsartán, un agente antihipertensivo inhibidor específico del receptor de angiotensina II subtipo AT1, en las membranas celulares del músculo liso vascular. Objetivo: Evaluar la bioequivalencia de dos productos farmacéuticos cuyo principio activo es valsartán, con base en la comparación de las medidas farmacocinéticas de cantidad y velocidad (en términos de tiempo requerido, con que el valsartán alcanza la circulación sanguínea después de ser administrado por vía oral a 15 voluntarios. Metodología: Se realizó un estudio aleatorizado, cruzado, a doble ciego, de dosis única en 15 voluntarios sanos con edades entre 19 y 28 años. El estudio se realizó en dos períodos; en cada período de tratamiento se administró una dosis de 320 mg de valsartán, con un período de lavado de 8 días entre el primero y segundo tratamiento. Las concentraciones plasmáticas se evaluaron por HPLC/UV con el método de adición de estándar, empleando losartán como estándar interno. Resultados: Para las tabletas de valsartán se obtuvieron los siguientes parámetros farmacocinéticas: ABC de 44,893 µg/mlxh, Cmáx de 6,430.3 µg/ml y Tmáx de 2 h. Para las cápsulas de valsartán se obtuvieron los siguientes parámetros farmacocinéticas: ABC de 44,963 µg/mlxh, Cmáx de 5,831.4 µg/ml y Tmáx de 2.5 h. Conclusión: Los resultados del estudio no mostraron diferencias significativas en los niveles de concentraciones plasmáticas después de la administración de las dos formulaciones de valsartán: 80 mg tabletas y 80 mg cápsulas. Por tanto, el diseño del estudio y la aplicación de los protocolos escogidos permitieron demostrar la bioequivalencia entre los productos.

  4. Caracterização e identificação de cultivares e seleções de pereiras através de marcadores RAPD

    Directory of Open Access Journals (Sweden)

    SAWAZAKI HAIKO ENOK

    2002-01-01

    Full Text Available Trinta e seis acessos de pereira representando diversas espécies, híbridos e seleções do banco de germoplasma do Instituto Agronômico (IAC foram geneticamente caracterizados através de marcadores RAPD. Cada primer originou de 10 a 19 bandas, sendo que 26 deles forneceram 250 bandas polimórficas, de um total de 353. Os primers OPC02, OPC08, OPD02, OPD19, OPD20 e OPE06 revelaram bandas específicas para as peras orientais e OPA01, OPA11, OPC08, OPD04, OPD09 e OPD15 para as ocidentais. O dendograma obtido foi confirmado pela análise de coordenada principal, originando três principais agrupamentos: 1 Todas as pereiras lançadas pelo IAC, como 'Seleta', 'Triunfo', 'Primorosa', 'Tenra', IAC 16-41, 'Centenária', além de 'William's', 'Packham's Triumph', 'D'água', 'Hood', 'M. Sieboldt', 'Kieffer','Branca Francesa' e 'Schimidt'. 2 As pereiras asiáticas, como 'Okusankichi', 'Shinseiki', 'Atago', 'Hakko', 'Hosui', 'Nijiseiki', 'Kosui' e 'Ya-li', além de 'Nodji', 'Limeira' e todas as seleções IAC das séries 193; 293 e 393. 3 Todas as pereiras porta-enxertos da série Taiwan (P. calleryana D., além de 'Manshu Mamenashi' (P. betulaefolia B.. Evidenciou-se que os cultivares IAC possuem maior proximidade genética com as peras ocidentais (Pyrus communis L., mesmo sendo descendentes de 'Hood', material suspeito de ser híbrido interespecífico entre P. communis e P. serotina R.. Os resultados ratificaram a importância dos marcadores RAPD para a identificação de cultivares, seleções e híbridos pertencentes aos diferentes grupos botânicos, mostrando ser ferramenta de apoio adequada a programas de melhoramento genético de fruteiras.

  5. Hybrids between olive flounder Paralichthys olivaceus and stone flounder Kareius bicoloratus: karyotype, allozyme and RAPD analyses

    Science.gov (United States)

    You, Feng; Wang, Wei; Xu, Dongdong; Zhu, Xiangping; Ni, Jing; Wu, Zhihao; Xu, Yongli; Wang, Xincheng; Zhang, Peijun

    2009-05-01

    The hybrid between olive flounder Paralichthys olivaceus and stone flounder Kareius bicoloratus was produced by artificial insemination of olive flounder eggs with stone flounder sperm. Sinistral and dextral are two types of hybrid progeny after metamorphosis. Karyotypes of both hybrid flounders are the same as those of the two parental species. Of the 22 loci examined from 12 allozymes, 12 confirmed hybridization of the paternal and maternal loci in hybrids and no difference was found in allozyme patterns of sinistral and dextral hybrid fishes. RAPD patterns of these specimens were also studied with 38 primers selected from 104 tested. Among them, the PCR products of 30 primers showed hybridization of the paternal and maternal bands. Genetic variation between hybrids and their parental stocks was analyzed by RAPD using 10 of the above 38 primers. The average heterozygosity and genetic distance were calculated. The results suggested that the filial generation could inherit a little more genetic materials from paternal fish than that from maternal fish.

  6. Genetic diversity in mesoamerican populations of mahogany (Swietenia macrophylla), assessed using RAPDs.

    Science.gov (United States)

    Gillies, A C; Navarro, C; Lowe, A J; Newton, A C; Hernández, M; Wilson, J; Cornelius, J P

    1999-12-01

    Swietenia macrophylla King, a timber species native to tropical America, is threatened by selective logging and deforestation. To quantify genetic diversity within the species and monitor the impact of selective logging, populations were sampled across Mesoamerica, from Mexico to Panama, and analysed for RAPD DNA variation. Ten decamer primers generated 102 polymorphic RAPD bands and pairwise distances were calculated between populations according to Nei, then used to construct a radial neighbour-joining dendrogram and examine intra- and interpopulation variance coefficients, by analysis of molecular variation (AMOVA). Populations from Mexico clustered closely together in the dendrogram and were distinct from the rest of the populations. Those from Belize also clustered closely together. Populations from Panama, Guatemala, Costa Rica, Nicaragua and Honduras, however, did not cluster closely by country but were more widely scattered throughout the dendrogram. This result was also reflected by an autocorrelation analysis of genetic and geographical distance. Genetic diversity estimates indicated that 80% of detected variation was maintained within populations and regression analysis demonstrated that logging significantly decreased population diversity (P = 0.034). This study represents one of the most wide-ranging surveys of molecular variation within a tropical tree species to date. It offers practical information for the future conservation of mahogany and highlights some factors that may have influenced the partitioning of genetic diversity in this species across Mesoamerica.

  7. Genetic variability of Triatoma flavida and Triatoma bruneri (Hemiptera: Reduviidae by RAPD-PCR technique

    Directory of Open Access Journals (Sweden)

    Jorge Fraga

    2011-02-01

    Full Text Available The Triatominae (Hemiptera:Reduviidae contains the principal and potential Chagas disease vectors present in Mexico, Central America and South America. Triatoma flavida and T. bruneri are Cuban species. These species are closely related according to morphology and were considered synonyms until 1981, when they were separated on the grounds of external characters of the body and the morphology of male genitalia. The present study seeks to analyze genetic polymorphism of T. flavida and T. bruneri populations using RAPD techniques, and to assess the genetic relationship between these species. Ten random primers were used to evaluate the genetic variability among species using RAPD-PCR. The genetic flow among them was calculated. The dendrogram based on calculated Jaccard distances showed two clearly distinguishable clusters which coincided with the studied species. Within each species, moderate genetic differentiation (Fst 0.05-0.15 and migration rates (N > 1 were found among populations, that reveal gene flow and genetic homogeneity. Between species, the Fst value showed a high genetic differentiation and the migration rate was insufficient to maintain genetic homogeneity, and confirmed the absence of gene flow between them. Our results confirm the genetic variability among T. flavida and T. bruneri species.

  8. Genetic variability of Triatoma flavida and Triatoma bruneri (Hemiptera: Reduviidae) by RAPD-PCR technique.

    Science.gov (United States)

    Fraga, Jorge; Rodriguez, Jinnay; Fuentes, Omar; Hernández, Yenin; Castex, Mayda; Gonzalez, Raul; Fernández-Calienes, Aymé

    2011-01-01

    The Triatominae (Hemiptera:Reduviidae) contains the principal and potential Chagas disease vectors present in Mexico, Central America and South America. Triatoma flavida and T. bruneri are Cuban species. These species are closely related according to morphology and were considered synonyms until 1981, when they were separated on the grounds of external characters of the body and the morphology of male genitalia. The present study seeks to analyze genetic polymorphism of T. flavida and T. bruneri populations using RAPD techniques, and to assess the genetic relationship between these species. Ten random primers were used to evaluate the genetic variability among species using RAPD-PCR. The genetic flow among them was calculated. The dendrogram based on calculated Jaccard distances showed two clearly distinguishable clusters which coincided with the studied species. Within each species, moderate genetic differentiation (Fst 0.05-0.15) and migration rates (N > 1) were found among populations, that reveal gene flow and genetic homogeneity. Between species, the Fst value showed a high genetic differentiation and the migration rate was insufficient to maintain genetic homogeneity, and confirmed the absence of gene flow between them. Our results confirm the genetic variability among T. flavida and T. bruneri species.

  9. Genetic Diversity Evaluation of Maize Recurrent Selection Population with RAPD Marker

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    The genetic diversity of maize populations Wuxi (W) from Southwest China, BSSS9(B) from America, Mohuangjiu (M) from Mexico, WBMC0 synthesized by W, B, M as main parents,WBMC1 one cycle selected from WBMC0 were evaluated by RAPD molecular marker. The results showed that :(1) Totally 89 fragments (loci) were amplified by 15 10-mar random primers, the proportion of polymorphic loci were W 76. 4%, B 75. 3%, M 79. 8%,WBMC0 85. 4% and WBMC1 92. 1% respectively; (2) The mean gene heterozygosity based on 89 loci was W 0. 285, B 0. 252, M 0. 296, WBMC0 0. 327 and WBMC1 0. 346; (3) The mean genetic distance based on 89 loci were W 0. 2533, B 0. 2246, M 0. 2481, WBMC0 0. 3006 and WBMC1 0. 3119; (4) The genotypic mean numbers amplified by 15 primers were W 9.1, B 7.8, M 8.5, WBMC0 10. 1 and WBMC1 10. All indexes indicated that the synthesized maize population were more polymorphic than the parent populations in DNA level. One cycle selection did not reduce the variation. The new conception of "genotypic diversity" (the number of genotypes in a population) was provided to describe the genetic diversity for any population being equilibrium or unequilibrium in genetics. The principle and technical system were discussed for evaluating genetic variation of recurrent selection population using RAPD molecular marker.

  10. Genetic variation of natural and cultured stocks of Paralichthys olivaceus by allozyme and RAPD

    Institute of Scientific and Technical Information of China (English)

    YOU Feng; ZHANG Peijun; WANG Keling; XIANG Jianhai

    2007-01-01

    Population genetics of the left-eyed flounder, Paralichthys olivaceus, including natural and cultured stocks distributed in the coastal waters near Qingdao of eastern maritime China, was analyzed in allozyme and RAPD. The results showed that among total 29 gene loci of 15 isozymes, 9 and 7 were polymorphic in natural and cultured stocks, respectively. The status of genetic diversity in P olivaceus is low in terms of polymorphic loci in chi-square test and genetic departure index of Hardy-Weinberg equilibrium. More alleles in IDHP, CAT, GDH and Ldh-C allozymes were found in the fish, which could be used as markers in assortive breeding and distinguishing stock, population or species evolution. Total 88 and 86 RAPD bands ranging from 200 to 2 500 bp were recognized individually in average of 7.8-8.0 bands per primer. The genetic diversity in cultured stock is lower than that in natural ones showing an obviously decreasing genetic divergence. Therefore, effective countermeasures must be taken to protect genetic resources of marine cultured fishes. The 2 markers have their own pros and cons. Combining the 2 markers to investigate the genetic variation of populations is suggested. The results provide basic data of this flounder and they are useful for studying genetic improvement and genetic resources of the fish.

  11. Stem fasciation in cacti and succulent species--tissue anatomy, protein pattern and RAPD polymorphisms.

    Science.gov (United States)

    El-Banna, A N; El-Nady, M F; Dewir, Y H; El-Mahrouk, M E

    2013-09-01

    Fasciated and normal stem segments of Opuntia microdasys, Opuntia cylindrica, Huernia primulina and Euphorbia lactea were collected from the same plant and compared for their anatomy, water relations and genetic variations. Anatomical differences in terms of thickness of cuticle, vascular bundle, xylem and phloem were analyzed in both normal and fasciated stems. The mucilage cells were higher in the fasciated form of Opuntia microdasys than that in the normal form. Water status in terms of total water content (TWC), water deficit and relative water content (RWC) was influenced by fasciation. Genetic variations were tested in normal and fasciated stems using randomly amplified polymorphic DNA (RAPD) fingerprints and SDS-PAGE of soluble protein extracts. SDS-PAGE protein and RAPD analysis confirmed that normal and fasciated tissues were genetically different. Polymerase chain reaction (PCR) yielded different polymorphic banding patterns that were unique to each primer and distinguishable over all samples. The PCR results of normal and fasciated samples were significantly different in cases of primers P1, P2 and P3. These results indicate that occurrence of fasciation in Opuntia microdasys, Opuntia cylindrica, Huernia primulina and Euphorbia lactea is an epigenetic mutation of tissues.

  12. Molecular characterisation of a germplasm bank for Theobroma genus using the RAPD technique

    Directory of Open Access Journals (Sweden)

    Yovany Moreno

    2007-02-01

    Full Text Available RAPD markers (Random Amplified Polymorphic DNA were used for analysing 145 individuals (128 T. grandiflorum and 17 T. bicolor from the ex situ Theobroma genus germplasm bank at Instituto Sinchi, located at San José del Guaviare. 5 primers able to generated polymorphism were selected from an initial set of 20, generating 114 bands that enable to us to distinguish between more than 99% of individuals analysed: 57 bands for T. grandiflorum (84.2% polymorphic, 45 bands for T. bicolor (26.7% polymorphic and 12 bands shared between the two species (58.3% polymorphic. A high degree of intra-specific similarity particularly in T. bicolor was established from the similarity matrix obtained by using the Dice index and represented in a UPGMA dendrogram and the principal components analysis (PCA. The comparison of this analysis with a previous morpho-agronomic evaluation of some T. grandiflorum individuals revealed that the groups generated on the basis of its agronomic and morphological traits were heterogeneous at molecular level. The obtained information will be used as a tool in strategies regarding maintenance, enrichment and use of the germplasm bank. Key words: Theobroma grandiflorum, Theobroma bicolour, RAPD, molecular characterisation.

  13. Hybrids between olive flounder Paralichthys olivaceus and stone flounder Kareius bicoloratus: karyotype, allozyme and RAPD analyses

    Institute of Scientific and Technical Information of China (English)

    YOU Feng; WANG Wei; XU Dongdong; ZHU Xiangping; NI Jing; WU Zhihao; XU Yongli; WANG Xincheng; ZHANG Peijun

    2009-01-01

    The hybrid between olive flounder Paralichthys olivaceus and stone flounder Kareius bicoloratus was produced by artificial insemination of olive flounder eggs with stone flounder Sperm. Sinistral and dextral are two types of hybrid progeny after metamorphosis. Karyotypes of both hybrid flounders are the same as those of the two parental species. Of the 22 loci examined from 12 allozymes,12 confirmed hybridization of the paternal and matemal loci in hybrids and no difference was found in allozyme patterns of sinistral and dextral hybrid fishes. RAPD patterns of these specimens were also studied with 38 primers selected from 104 tested. Among them, the PCR products of 30 primers showed hybridization of the paternal and maternal bands. Genetic variation between hybrids and their parental stocks was analyzed by RAPD using 10 of the above 38 primers. The average heterozygosity and genetic distance were calculated. The results suggested that the filial generation could inherit a little more genetic materials from paternal fish than that from maternal fish.

  14. Evaluation of genetic variation in the clown Knifefish, Chitala chitala, using allozymes, RAPD, and microsatellites.

    Science.gov (United States)

    Mandal, Anup; Lal, Kuldeep Kumar; Mohindra, Vindhya; Singh, Rajeev Kumar; Punia, Peyush; Chauhan, U K; Lakra, Wazir Singh

    2009-04-01

    Twenty-seven enzyme systems, six random amplified polymorphic DNA (RAPD) primers, and two microsatellite loci were tested to determine intraspecific divergence in the natural population of the endangered Indian featherback fish, Chitala chitala, for the first time. The 262 samples of C. chitala were collected from six riverine locations in India: the Satluj, Ganga (Ghagra, Bhagirathi, and Brahmaputra), Mahanadi, and Narmada river systems. The analysis revealed population subdivisions, with an F(ST) value from 0.1235 (95% confidence 0.0868-0.1621) for RAPD and a combined F(ST) of 0.0344 (95% confidence 0.0340-0.0350) for microsatellite loci. An analysis of 38 allozyme loci did not reveal any polymorphism in the samples from any of the riverine localities; a possible explanation for this could be that the ancestors of Chitala could have faced a population reduction in prehistoric periods, as low allozyme variation is also reported for other species of Chitala from south Asia.

  15. Genetic diversity of Cosmos species revealed by RAPD and ISSR markers.

    Science.gov (United States)

    Rodríguez-Bernal, A; Piña-Escutia, J L; Vázquez-García, L M; Arzate-Fernández, A M

    2013-12-04

    The genus Cosmos is native of America and is constituted by 34 species; 28 of them are endemic of Mexico. The cosmos are used as a nematicide, antimalarial, and antioxidative agent. The aim of this study was to estimate the genetic diversity among 7 cosmos species based on random amplified polymorphic DNA (RAPD) and inter-simple sequences repeats (ISSR) markers. With RAPD markers, the obtained polymorphism was 91.7 % and the genetic diversity was 0.33, whereas these values were 65.6%, and 0.22 from ISSR markers, respectively, indicating the presence of high genetic diversity among the Cosmos species that were analyzed. The unweighted pair group method with arithmetic mean dendrograms that were obtained with both markers were notably similar, revealing 2 clusters and indicating a clear genetic differentiation among the Cosmos species that were assessed. The first cluster comprised the species Cosmos sulphureus, Cosmos pacificus, and Cosmos diversifolius, while the second cluster included the species Cosmos purpureus, Cosmos crithmifolius, Cosmos bipinnatus, and Cosmos parviflorus. Besides this, the Cosmos species were clustered according to their collection sites. The Mantel test corroborates the correlation between the genetic distance and the geographic altitude of each Cosmos species. The results suggest that it is necessary to preserve the Cosmos species in their natural habitat in addition to the germoplasm collection for ex situ conservation.

  16. Genetic Diversity of Testa Pigments and RAPD Marker of Yellow-Seeded Rapeseed (Brassica napus L.)

    Institute of Scientific and Technical Information of China (English)

    ZHANG Xue-kun; CHEN Li; YIN Jia-ming; TANG Zhang-lin; LI Jia-na

    2003-01-01

    14 yellow-seeded rapeseed lines (Brassia napus L. ) from different genetic sources were used toanalyze diversity of testa pigments content, oil and protein content, and RAPD markers. The results showedthat the anthocyanin and melanin were the most important pigments in testa and their content were responsiblefor the variation in seed color ranging from orange to black yellow, 14 yellow-seeded lines could be classifiedinto 3 groups., high anthocyanin content group with anthocyanin content over 2.54 mg g-1 DW, the seed colorwas light yellow or orange; low pigments content group with low content of anthocyanin and melanin, the testawas transparent and the seed color was light yellow, greenish yellow or twany; high melanin content groupwith melanin content over 178.4U(A290nm), the testa was black, the seed color was black yellow. Oil eantentchanged from 36.2% to 45.5%, protein content from 21.1% to 27.7%, and the correlation analysis revealedthat the oil content is highly significantly negatively correlated with the protein content. The cluster analysisshowed that the extensive genetic variation existed among 14 yellow-seeded lines by using unweighted pairedgroup method with arithmetic average (UPGMA) based on RAPD markers which were amplified with decamerprimers, the genetic similarity among them ranged from 0.25 to 0. 909, and 14 yellow-seeded lines could putinto 2 clusters corresponding to genome difference.

  17. Photodynamic Treatment versus Antibiotic Treatment on Helicobacter pylori Using RAPD-PCR

    Science.gov (United States)

    El-Batanouny, M. H.; Amin, R. M.; Ibrahium, M. K.; El Gohary, S.; Naga, M. I.; Salama, M. S.

    2009-09-01

    Helicobacter pylori is one of the most common causes of chronic bacterial infections in humans and is important in the pathogenesis of gastrointestinal disease, such as duodenal ulcer, gastric ulcer, Gastric adenocarcinoma, and lymphoma. Gastric adenocarcinoma remains one of the leading causes of cancer death in the world. The objective of this study was to assess the effect of photodynamic treatment and medication treatment of Helicobacter pylori using RAPD-PCR. The lethal photosensitization effect was determined by mixing suspensions of H.pylori with Toluidine blue O (TBO) and plating out on blood agar before irradiation with Helium neon (He-Ne) 632.8 nm. The susceptibility of Helicobacter pylori isolates to metronidazole and azithromycin were examined by E-test. Nine random primers were used to screen genetic polymorphism in DNA of different H.pylori groups. Six of them produced RAPD products while three failed to generate any product. The resulting data showed that, although the overall genetic differences between control groups and laser treated groups was higher than that between control groups and azithromycin treated groups yet it still law genetic variability. The main cause of cell death of PDT using TBO as a photosensitizer was mainly cell wall and cytoplasmic membrane.

  18. Estimation of the genetic diversity in tetraploid alfalfa populations based on RAPD markers for breeding purposes.

    Science.gov (United States)

    Nagl, Nevena; Taski-Ajdukovic, Ksenija; Barac, Goran; Baburski, Aleksandar; Seccareccia, Ivana; Milic, Dragan; Katic, Slobodan

    2011-01-01

    Alfalfa is an autotetraploid, allogamous and heterozygous forage legume, whose varieties are synthetic populations. Due to the complex nature of the species, information about genetic diversity of germplasm used in any alfalfa breeding program is most beneficial. The genetic diversity of five alfalfa varieties, involved in progeny tests at Institute of Field and Vegetable Crops, was characterized based on RAPD markers. A total of 60 primers were screened, out of which 17 were selected for the analysis of genetic diversity. A total of 156 polymorphic bands were generated, with 10.6 bands per primer. Number and percentage of polymorphic loci, effective number of alleles, expected heterozygosity and Shannon's information index were used to estimate genetic variation. Variety Zuzana had the highest values for all tested parameters, exhibiting the highest level of variation, whereas variety RSI 20 exhibited the lowest. Analysis of molecular variance (AMOVA) showed that 88.39% of the total genetic variation was attributed to intra-varietal variance. The cluster analysis for individual samples and varieties revealed differences in their population structures: variety Zuzana showed a very high level of genetic variation, Banat and Ghareh were divided in subpopulations, while Pecy and RSI 20 were relatively uniform. Ways of exploiting the investigated germplasm in the breeding programs are suggested in this paper, depending on their population structure and diversity. The RAPD analysis shows potential to be applied in analysis of parental populations in semi-hybrid alfalfa breeding program in both, development of new homogenous germplasm, and identification of promising, complementary germplasm.

  19. Genetic diversity of different accessions of Thymus kotschyanus using RAPD marker

    Directory of Open Access Journals (Sweden)

    Ahmad Ismaili

    2014-11-01

    Full Text Available Analysis of genetic diversity is a major step for understanding evolution and breeding applications. Recent advances in the application of the polymerase chain reaction make it possible to score individuals at a large number of loci. The RAPD technique has been successfully used in a variety of taxonomic and genetic diversity studies. The genetic diversity of 18 accessions of Thymus kotschyanus collected from different districts of Iran has been reported in this study, using 30 random amplified polymorphic DNA primers. Multivariate statistical analyses including principal coordinate analysis (PCOA and cluster analysis were used to group the accessions. From 29 primers, 385 bands were scored corresponding to an average of 13.27 bands per primer with 298 bands showing polymorphism (77.40%. A dendrogram constructed based on the UPGMA clustering method revealed three major clusters. The obtained results from grouping 18 accessions of T. kotschyanus with two studied methods indicated that in the most cases the applied methods produced similar grouping results. This study revealed nearly rich genetic diversity among T. kotschyanus accessions from different regions of Iran. The results showed RAPD marker was a useful marker for genetic diversity studies of T. kotschyanus and it was indicative of geographica variations.

  20. RANDOMLY AMPLIFIED POLYMORPHIC DNA (RAPD FINGERPRINTING OF SIX INDONESIAN POPULATIONS OF GIANT FRESHWATER PRAWN, Macrobrachium rosenbergii

    Directory of Open Access Journals (Sweden)

    Imron Imron

    2009-12-01

    Full Text Available Indonesia is rich of giant fresh water prawn (GFP germ plasms. Best utilization of these resources for the purpose of either aquaculture development or conservation of genetic resources requires some information on the structure and levels of their genetic diversity. This study was aimed to characterize those GFP genetic resources by applying RAPD genetic markers. Six Indonesian populations of GFP from Asahan, Barito, Ciasem, Ogan, GImacro and Papua were collected and analyzed for their genetic variation using five RAPD primers. The results showed the diversity within the populations, as revealed by the level of polymorphism, ranged from 29% to 76% while genetic divergence between populations as shown by genetic distance ranged from 0.04 to 0.50. In terms of genetic divergence, two genetically distinct groups of GFP, namely the Papua GFP in one group and the remaining five GFP populations in the other, were identified. The results also showed the presence of specific population markers that are useful for genetic identification of GFP populations. Implication of these finding with regard to breed development is discussed.

  1. Relationships between some Thai cultivars of pineapple (Ananas comosus revealed by RAPD analysis

    Directory of Open Access Journals (Sweden)

    Siam Popluechai

    2007-11-01

    Full Text Available RAPD analysis of nine Thai pineapple cultivars, including 'Phulae', 'Sawee', 'Tradsithong', 'Phuket', 'Pattavia', 'Intrachitdang', 'Intrachitkhow', 'Petburi No.1', and 'Nanglae', showed that, of 40 arbitrary 10- mer primers, 17 primers gave 206 DNA fragments ranging from 510 to 4,700 bp. One hundred and forty-five (70.4% of the amplified fragments were polymorphic. RAPD analysis using NTSYS-pc Version 2.01e also showed that the similarity coefficients among the cultivars were 0.643-0. 963. The dendrogram indicated that the cultivars were clustered into 3 groups, consistent with the morphological data. The first group, consisting of 'Phuket', 'Phulae', 'Tradsithong', 'Sawee', and 'Petburi No.1', had morphological characteristics of the Queen group, while those of the second ('Intrachitdang' and 'Intrachitkow' and the third ('Nanglae' and 'Pattavia' groups could be determined morphologically to be members of the Spanish and Cayenne groups, respectively. 'Intrachitdang' and 'Intrachitkow' have similarity coefficient of 0.963, while that of 'Phulae' and 'Phuket' is 0.950. These pairs of cultivars are probably the same cultivars. The morphological differences between them are probably caused by mutations, differences in environment and agricultural practices, or combinations of these factors.

  2. Genetic variation and phylogenetic relationships in oil palm (Elaeis guineensis Jacq. based on RAPD analysis

    Directory of Open Access Journals (Sweden)

    Nualsri, C.

    2005-05-01

    Full Text Available The genetic variability and phylogenetic relationships in oil palm (Elaeis guineensis Jacq. were studied using RAPD (Random Amplified Polymorphic DNA. Leaf samples of 151 plants were collected from different areas in southern Thailand. DNA from the leaf samples was isolated using CTAB buffer and screened by decamer oligonucleotide primers. Among the total of 160 primers screened, 7 primers (OPB-08, OPR-11, OPT-06, OPT-19, OPAB-01, OPAB-09 and OPAB-14 were chosen to analyse for genetic variation in 151 individuals representing 52 dura, 60 tenera and 39 pisifera. Two hundred and nine amplified fragments were obtained from 7 primers with an average of 29.85 RAPD markers per primer. A dendrogram showing genetic similarities among oil palm was constructed based on polymorphic bands using UPGMA (Unweighted Pair-Group Method Using Arithmetic Average. Cluster analysis was performed using the SPSS program, which revealed four major clusters: 1 dura, tenera and pisifera from Paorong Oil Palm Company, Oil Palm Research Center, dura and tenera from private plantation in Krabi, and dura from Thepa Research Station;2 dura and tenera from Thai Boonthong Company, pisifera and tenera from Thepa Research Station, dura, tenera and pisifera from Klong Hoi Khong Research Station; 3 and 4 dura and tenera from Univanit Company, respectively. In general, a similarity index showed relatively high levels of 0.6 or greater.

  3. Establishing a diagnostic system for detecting Ralstonia solanacearum and genetic differentiation using RAPD molecular markers

    Directory of Open Access Journals (Sweden)

    Edisson Chavarro Mesa

    2007-02-01

    Full Text Available A polymerase chain reaction-based diagnostic test (PCR has been developed for amplifying a región and obtaining a 292 bp product by using specific 16S rDNA primers for the rapid and precise identification of the causative agent (Ralstonia solanacearum of bacterial withering of potato in asymptomatic tubers. The bacteria was isolated from potato tubers and banana fruit using culturing techniques and immunological and molecular ELISA-NCM and PCR tests, respectively. PCR detected the presence of R. solanacearum on asymptomatic tubers by contrast with ELISA-NCM which did not detect this pathogen. Analysing random amplified polymorphic DNA (RAPD led to differentiating and grouping R. solanacearum by geographical región and bacterial strain, suggesting that differences exist amongst existing collections according to their place of origin, presenting high genetic variability. The results showed that PCR is a sensitive and specific test for detecting R. solanacearum and can therefore be implemented as a method for controlling this pathogen in seed production and certification programmes in áreas free of the disease. The pathogen has been shown to be genetically heterogeneous according to the samples' geographical área thereby hampering control in áreas of Colombia experiencing phytosanitary problems with R. solanacearum in potato crops Key words: bacterial withered, moko, PCR-16S rADN, ELISA-NCM, PCR-RAPD.

  4. Genetic and alkaloid analysis of Menispermum dauricum DC. by RAPD and HPLC.

    Science.gov (United States)

    Zhou, Jue; Qu, Fan; Nan, Rui

    2007-01-01

    The aim of the present work was to determine whether dauricine could be used as a taxonomic marker for Menispermum dauricum DC., and to explore the correlation among RAPD, ecological markers and chemical markers. To this end, the chemical and genetic differences of 173 individual samples of M. dauricum from nine different sources were studied based on the relevant ecological factors including longitude, latitude, annual precipitation, mean temperature, annual accumulated temperature and mean sea level. The contents of dauricine in the sample rhizomes were assayed by HPLC with photodiode array detection. The leaves from the same sample were assayed using randomly amplified polymorphism DNA (RAPD). The genetic distances were then compared. Hierarchical cluster analysis and multiple linear stepwise regression analysis were used in the statistical analysis. The results indicated that the contents of dauricine were respectively correlated with the genetic distance (r = 1.000), longitude (r = 0.849), latitude (r = 0.861), annual precipitation (r = 0.903), mean temperature(r = 0.912), annual accumulated temperature (r = 0.919) and mean sea level (r = 0.925). It is concluded that the content of dauricine in M. dauricum is significantly correlated with genetic distance and ecological factors, and may be used as the taxonomic marker.

  5. Genetic similarity of Brazilian hull-less and malting barley varieties evaluated by RAPD markers Similaridade genética de variedades brasileiras de cevada nua e cevada cervejeira avaliada por marcadores RAPD

    Directory of Open Access Journals (Sweden)

    Mariela Aparecida Kroth

    2005-01-01

    Full Text Available Barley (Hordeum vulgare L. is widely used for brewing and animal feed. Recently, it has become desirable for human consumption due to its high nutritional significance, specially hull-less or naked barley. There are differences in nutritional and malting characteristics among barley varieties. RAPD procedure is able to separate barley varieties at various similarity levels. The aim of this work was the RAPD analysis of six Brazilian hull-less varieties and seven malting varieties. PCR reactions were performed with eleven random primers. A total of 34 RAPD fragments was obtained with five primers. A dendrogram was constructed based on the Jaccard similarity coefficient. Barley varieties displayed an average similarity coefficient of 0.53. Intravarietal monomorphic fragments allowed differentiation among varieties. The averages of intravarietal similarity coefficients ranged from 0.53 to 0.85. RAPD markers, detected in this work, were suitable for differentiation among Brazilian barley varieties.A cevada (Hordeum vulgare L. é amplamente empregada na produção de cerveja e na ração animal. Recentemente, este cereal tornou-se desejável na alimentação humana devido ao seu alto valor nutricional, principalmente da cevada nua ou sem casca. Existem diferenças nas características nutricionais e para malteação entre as diversas variedades de cevada. O procedimento RAPD é capaz de separar as variedades de cevada em vários níveis de similaridade. O objetivo desse trabalho foi analisar por RAPD seis variedades brasileiras de cevada nua e sete variedades de cevada cervejeira. Reações de PCR foram realizadas com onze iniciadores aleatórios. Um total de 34 fragmentos de amplificação foi obtido com cinco destes iniciadores. Baseado no coeficiente de similaridade de Jaccard, um dendrograma foi construído. As variedades de cevada apresentaram índice médio de similaridade de 0,53. Fragmentos monomórficos intravariedades permitiram a

  6. Evaluation of RAPD-PCR and protein profile analysis to differentiate Vibrio harveyi strains prevalent along the southwest coast of India

    Indian Academy of Sciences (India)

    Biswajit Maiti; Malathi Shekar; Rekha Khushiramani; Iddya Karunasagar; Indrani Karunasagar

    2009-12-01

    Sixty five isolates of Vibrio harveyi were subjected to random amplified polymorphic DNA (RAPD)-PCR analysis and protein profiling to investigate the genetic variability among V. harveyi prevalent along the coast and also assess the discriminating ability of these two molecular methods. A total of 10 RAPD primers were assayed for their specificity in detecting V. harveyi, of which only two primers: PM3 and CRA25 were highly reproducible and found suitable for use in RAPD-PCR. The genetic diversity among V. harveyi isolates assessed by RAPD-PCR using PM3 primer yielded 35 different RAPD patterns which clustered the isolates into 15 groups at 72% similarity level. Similarly, RAPD-PCR with CRA25 clustered the 38 patterns into 10 groups at 74% similarity. The discriminatory index $(D)$ value calculated for RAPD fingerprints generated with PM3 and CRA25 were 0.90 and 0.85, respectively. On the other hand, molecular typing of V. harveyi using whole cell proteins generated profiles that showed no major difference indicating the technique to be not useful in typing strains of this bacterium. However, a few of the isolates showed the presence of unique band of 28 kDa that needs to be further investigated to understand the role of the protein in disease process if any.

  7. Random amplified polymorphic DNA analysis of effective Rhizobium sp. associated with beans cultivated in brazilian cerrado soils Caracterização, via RAPD, de estirpes efetivas de Rhizobium sp associadas ao feijoeiro cultivado em solos de cerrado no Brasil

    Directory of Open Access Journals (Sweden)

    Isnia Aparecida de Oliveira

    2000-03-01

    Full Text Available Efficient bean nodulating Rhizobium strains, isolated from different Brazilian cerrado soils, were characterized by RAPD. This study showed great genetic heterogeneity among R. tropici and R. leguminosarum bv. phaseoli strains and allowed the constitution of genetic clusters, besides indicating the most suitable primers for this characterization. The groups of genetically distinct strains can be used in competitiveness studies to select appropriate Rhizobium strains for bean inoculation in cerrado soils.Estirpes de Rhizobium eficientes na nodulação do feijoeiro, isoladas de diferentes solos da região do cultivo dessa leguminosa nos cerrados brasileiros, foram caracterizadas via RAPD. Esse estudo mostrou grande heterogeneidade genética entre as estirpes de R. tropici e R. leguminosarum bv. phaseoli testadas e permitiu a definição de grupos genéticos, além de indicar os "primers" mais adequados para essa caracterização. Os grupos de estirpes geneticamente distintas podem ser usados em estudos de competitividade, importantes para obtenção de resultados positivos na inoculação dessa leguminosa em solos de cerrado.

  8. ISSR and RAPD based evaluation of genetic stability of encapsulated micro shoots of Glycyrrhiza glabra following 6 months of storage.

    Science.gov (United States)

    Mehrotra, Shakti; Khwaja, O; Kukreja, A K; Rahman, L

    2012-11-01

    In vitro grown axillary micro shoots of Glycyrrhiza glabra were encapsulated in alginate beads. Following 6 months of normal storage at 25 ± 2°C the re growth of encapsulated G. glabra micro shoots, reached 98% within 30 days of incubation on MS medium supplemented with 0.1 mg/l IAA. Re growth was characterized by the development of both shoot and root from single encapsulated micro shoot. Healthy plants were established to glass house with 95% survival. The genetic fidelity of plants obtained after conversion of alginate beads was ascertained through 10 RAPD and 13 ISSR primers. Of the 10 RAPD primers tested, 6 of them produced 14 clear and reproducible amplicons with an average of 2.3 bands per primer out of which 28.57% were polymorphic generated by only two primers. Eight ISSR primers produced total 37 bands ranging between 300 and 3,500 bp length. Number of scorable bands for each primer varied from 3 to 8 with an average of 4.6 bands per primer. Cluster analysis from ISSR and RAPD showed that all the tested plants including the mother plant distributed in two major groups with similarity coefficient ranging from 0.91 to 0.96 for RAPD and 0.89 to 0.97 for ISSR.

  9. Identification of Leishmania Species Isolated from Human Cutaneous Leishmaniasis, using Random Amplified Polymorphic DNA (RAPD-PCR

    Directory of Open Access Journals (Sweden)

    H Hajjaran

    2004-10-01

    Full Text Available This cross-sectional study was designed to isolate of Leishmania spp from cutaneous leishmaniasis patients and characterized them by RAPD-PCR technique. Eighty- seven Leishmania isolates from 112 samples were collected from cutaneous leishmaniasis (CL patients who referred to Mashhad Health Centers from August 2002 to May 2004. Desirable samples (87 isolates were characterized by RAPD-PCR method using four selected oligoprimers. Electrophoresis patterns from each isolate were compared with reference strains of L. major, L. tropica and L. infantum. The results showed that 94.2% and 5.8% of isolates were similar to L.tropica and L.major reference strain, respectively. Four isolates that were determined by RAPD-PCR as L.major, could produce ulcer at the base tail of BALB/c mice, 4 - 12 weeks after inoculation but none of L. tropica isolates produced any lesions at the site of injection in the animals. The results indicate that L. tropica species are dominant in the studied areas of Mashhad city and RAPD-PCR technique is a suitable tool for Leishmania characterization in epidemiological studies.

  10. Genetic Characterization by RAPD Analysis of Isolates of Fusarium oxysporum f. sp. erythroxyli Associated with an Emerging Epidemic in Peru.

    Science.gov (United States)

    Nelson, A J; Elias, K S; Arévalo G, E; Darlington, L C; Bailey, B A

    1997-12-01

    ABSTRACT An epidemic of vascular wilt caused by Fusarium oxysporum f. sp. erythroxyli is currently occurring on Erythroxylum coca var. coca in the coca-growing regions of the Huallaga Valley in Peru. Random amplified polymorphic DNA (RAPD) analysis of isolates of the pathogen was undertaken to elucidate its genetic complexity, as well as to identify a specific DNA fingerprint for the pathogen. Two hundred isolates of Fusarium were collected from 10 coca-growing regions in Peru. Of these, 187 were confirmed to be F. oxysporum, and 143 of the F. oxysporum were shown to be pathogens of coca by a root-dip pathogenicity test. The pathogens could be grouped into two subpopulations based on RAPD analysis, and no polymorphism in RAPD pattern was observed among isolates of either subpopulation. Both subpopulations were present in the central Huallaga Valley, where earliest reports of the epidemic occurred. RAPD analysis could easily distinguish the isolates of F. oxysporum f. sp. erythroxyli from the nonpathogenic isolates of F. oxysporum from E. coca var. coca, indicating its utility in DNA fingerprinting.

  11. CARACTERIZAÇÃO DA VARIABILIDADE GENÉTICA EM COUVE-MANTEIGA UTILIZANDO ISOENZIMAS E RAPD

    Directory of Open Access Journals (Sweden)

    SAWAZAKI HAIKO ENOK

    1997-01-01

    Full Text Available Estudou-se a variabilidade genética em couve (Brassica oleracea L. var. acephala D.C. tipo manteiga por intermédio do polimorfismo enzimático em gel de poliacrilamida e do polimorfismo de DNA, denominado RAPD (Random Amplified Polymorphic DNA, com base na amplificação de segmentos de DNA ao acaso. Avaliaram-se quinze clones de couve-manteiga do Banco Ativo de Germoplasma do Instituto Agronômico (IAC, utilizando-se extratos de folhas para análise de isoenzimas e marcador RAPD com os "primers" dos kits A e B da Operon Technologies. Entre as isoenzimas estudadas, as mais polimórficas foram as fosfoglucomutase (PGM, peroxidase (PRX e esterase (EST, tendo o sistema PGM realizado a melhor caracterização. Verificou-se a ocorrência de variabilidade genética por meio de isoenzimas e RAPD, porém não foi observada a similaridade entre os dendrogramas obtidos por ambos os tipos de marcadores, sugerindo que as isoenzimas forneceram menos informação sobre o genoma. A maior eficácia do RAPD foi devida à possibilidade de processar maior número de análises, evidenciando mais detalhes sobre o genoma.

  12. Molecular characterization of Pasteurella multocida isolates obtained from poultry, ruminant, cats and dogs using RAPD and REP-PCR analysis

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    Hesamaddin Shirzad-Aski

    2016-09-01

    Full Text Available In the present study, Randomly Amplified Polymorphic DNA‌ (RAPD and Repetitive Extragenic Palindromic sequence-based Polymerase Chain Reaction (REP-PCR were used to characterize 131 isolates of Pasteurella multocida, originating from different healthy and diseased animal species obtained from several geographical regions of Iran. The RAPD and REP-PCR generated amplified products in the range of 300 to 3400 bp and 200 to 2850 bp, respectively. Among all of the P. multocida isolates, cluster analysis revealed that 63 clusters and nine untypable isolates and 81 clusters and six untypable isolates were produced with RAPD and REP-PCR methods, respectively. The results indicated that the REP-PCR method showed a slightly higher level of discrimination power in differentiating of P. multocida isolates as compared with RAPD. The results showed that a considerable level of genetic diversity exists among P. multocida isolates even in the isolates with the same animal or geographical origins. There was no host- and region-specific pattern. In addition, the isolates obtained from the healthy and diseased animal did not reveal any correlation genotypic profiles, which could be supported by the hypothesis that P. multocida is a strictly opportunistic pathogen. In conclusion, because of a large amount of genetic heterogeneityin the P. multocida isolates, Pasteurellosis may be caused by different clones in the same herd or animal.

  13. « El azar y la necesidad » Élite y elecciones matrimoniales en Buenos Aires 1776-1820. Las familias Azcuénaga y Andonaegui

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    Maria Selva Senor

    2005-01-01

    Full Text Available En el presente articulo se reconstruyen las trayectorias matrimoniales de dos familias de la élite en el Río de la Plata entre 1776 y 1810 con el objeto de descubrir las estrategias seguidas por las mismas y las repercusiones del azar. Desde una perspectiva microanalitica y a partir de la utilización de la noción de redes sociales se intentará ilustrar acerca de las reglas sociales en una sociedad con variados contextos normativos superpuestos, que incluyen asimismo a la política del Estado y en la que los agentes pueden manipular reglas y tomar decisiones haciendo que el grado de incertidumbre sea mayor.

  14. A retrospective study of intravenous sodium stibogluconate alone and in combinations with allopurinol, rifampicin, and an immunomodulator in the treatment of Indian post-kala-azar dermal leishmaniasis

    Directory of Open Access Journals (Sweden)

    Ramesh V

    2010-01-01

    Full Text Available Background and Aims: A retrospective analysis of treatment outcome using recommended dose of sodium stibogluconate (SSG alone and in combination with other antileishmanial drugs in adults with post-kala-azar dermal leishmaniasis (PKDL attending as outpatients. Methods: A total of 61 patients seen over ten years were included in the report. All had polymorphic lesions. Diagnosis was based on clinical picture, hailing from kala-azar (KA endemic area, exclusion of other dermatoses, histopathology, and therapeutic response. Patients were distributed into two groups: Group I (n = 32, where SSG was given intravenously; in Group II (n = 29, they were allocated to one of four categories using SSG in combination with other drugs. In the first category, SSG was given along with allopurinol (n = 10; in second with rifampicin (n = 6; and in third with both allopurinol and rifampicin (n = 5. In the fourth category, SSG was administered with an immunomodulator (n = 8, Mw vaccine, known to enhance host Th1 response. Results: Only 12 out of 61 patients completed treatment till histopathologic evidence of cure, five in Group I and seven in Group II, no patient being from third category. None had taken SSG without interruptions. Time taken for papulonodules to subside was similar in both groups, but erythema and induration subsided earlier in Group II. Group I patients attained cure after 120 injections while in Group II it took 95 injections in SSG + allopurinol and Mw vaccine categories respectively, and 110 with SSG + rifampicin. Nevertheless this was insufficient to facilitate compliance. Poor performance and high dropouts related to long duration of therapy, thrombophlebitis, difficulty in accessing veins, disabling rheumatic side-effects and practical problems. Liver, renal and pancreatic functions and ECG remained normal. Conclusion: No major advantage was obtained using allopurinol, rifampicin or Mw vaccine along with SSG as compared to SSG alone.

  15. Inheritance of rapeseed (Brassica napus)-specific RAPD markers and a transgene in the cross B.juncea x (B.juncea x B.napus)

    DEFF Research Database (Denmark)

    Frello, S.; Hansen, K.R.; Jensen, J.

    1995-01-01

    , with B. juncea as the female parent, was successful both in controlled crosses and spontaneously in the field. The controlled backcrossing of selected hybrids to B. juncea, again with B. juncea as the female parent, also resulted in many seeds. The BC1 plants contained from 0 to 20 of the rapeseed RAPD...... markers, and the frequency of inheritance of individual RAPD markers ranged from 19% to 93%. The transgene was found in 52% of the plants analyzed. Five synteny groups of RAPD markers were identified. In the hybrids pollen fertility was 0-28%. The hybrids with the highest pollen fertility were selected...

  16. Genetic and conservation of Araucaria angustifolia: III DNA extraction protocol and informative capacity of RAPD markers for the analysis of genetic diversity in natural population

    OpenAIRE

    2004-01-01

    This study was aimed at adapting a DNA extraction protocol by Araucaria angustifolia leaves, and testing the informative capacity of RAPD markers for genetics diversity analysis in natural populations of this species. The extraction method was standardized by eight tested protocols and it was possible to obtain good quality DNA for RAPD reactions. The OD260/OD280 ratio ranged from 1.7 to 2.0 in 80% of the samples, indicating that they had a low level of protein contamination. The RAPD markers...

  17. Genetic diversity of Bemisia tabaci (Genn. Populations in Brazil revealed by RAPD markers

    Directory of Open Access Journals (Sweden)

    L.H.C. Lima

    2002-01-01

    Full Text Available Bemisia tabaci (Genn. was considered a secondary pest in Brazil until 1990, despite being an efficient geminivirus vector in beans and soybean. In 1991, a new biotype, known as B. tabaci B biotype (=B. argentifolii was detected attacking weed plants and causing phytotoxic problems in Cucurbitaceae. Nowadays, B. tabaci is considered one of the most damaging whitefly pests in agricultural systems worldwide that transmits more than 60 different plant viruses. Little is known about the genetic variability of these populations in Brazil. Knowledge of the genetic variation within whitefly populations is necessary for their efficient control and management. The objectives of the present study were to use RAPD markers (1 to estimate the genetic diversity of B. tabaci populations, (2 to study the genetic relationships among B. tabaci biotypes and two other whitefly species and (3 to discriminate between B. tabaci biotypes. A sample of 109 B. tabaci female individuals obtained from 12 populations in Brazil were analyzed and compared to the A biotype from Arizona (USA and B biotype from California (USA and Paraguay. Trialeurodes vaporariorum and Aleurodicus cocois samples were also included. A total of 72 markers were generated by five RAPD primers and used in the analysis. All primers produced RAPD patterns that clearly distinguished the Bemisia biotypes and the two other whitefly species. Results also showed that populations of the B biotype have considerable genetic variability. An average Jaccard similarity of 0.73 was observed among the B biotype individuals analyzed. Cluster analysis demonstrated that, in general, Brazilian biotype B individuals are scattered independently in the localities where samples were collected. Nevertheless, some clusters were evident, joining individuals according to the host plants. AMOVA showed that most of the total genetic variation is found within populations (56.70%, but a significant portion of the variation is found

  18. Tagging RAPD markers to a bacterial blight resistance gene in rice

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    @@The somaclonal mutant HX_3 has shown a broad spectrum resistance to bacterial blight. To study the inheritance of the bacterial blight resistance in HX_3, a cross was made between HX_3 and a susceptible cultivar Longtefu A. The F2 population of 418 plants was inoculated with Chinese bacterial blight strain Zhe 173 (pathotype Ⅳ ). Results showed that the F2 progenies segregated in a ratio of 3R∶ 1S (324 resistant plants and 94 susceptible plants). From the plants tested, 114 individuals (86 resistant and 28 susceptible) were chosen randomly for RAPD analysis. Twelve highly resistant and 12 highly susceptible plants were selected to form a resistant pool and a susceptible pool, respectively.

  19. Genetic linkage map of Brassica campestris L.using AFLP and RAPD markers

    Institute of Scientific and Technical Information of China (English)

    卢钢; 陈杭; 等

    2002-01-01

    A genetic linkage map comprised of 131 loci was constructed with an F2 population derived from an inter-subspecific cross between Brassica campestris L.ssp.chinensis cv.aijiaohang” and ssp.rapifera cv.,”'isihai”.The genetic map included 93 RAPD loci,36 AFLP loci and 2 morphological loci organized into 10 main linkage groups(LGs) and 2 small groups,covering 1810.9cM with average distance between adjacent markers being approximately 13.8cM.The map is suitable for identification of molecular markers linked to important agronomic traits.QTL analysis,and even for marker-assisted selection in breeding programs of Chinese cabbage and turnip.

  20. Detecting RAPD Markers Linked to Ripe Rot Resistance Genes in Chinese Wild Vitis

    Institute of Scientific and Technical Information of China (English)

    WANG Yue-jin; XU Yan; ZHANG Jian-xia; ZHOU Peng; WAN Yi-zhen

    2002-01-01

    With F1 individuals of the cross combination 88-110 of 83-4-96 ( V. quinquangularis Rehd. )× Muscat Rose ( V. vinifera L. ), the RAPD marker OPC15-1300 linked to ripe rot ( Gloeosporium fruetigenum Berk. ) resistance genes in Chinese wild Vitis was gained using bulked segregation analysis(BSA). And it was found that OPC15-1300 could be hereditary from the resistant parent (83-4-96) after the marker was tested in 50 F1 plants of the cross combination 88-110, 32 accessions of 8 Chinese wild Vitis species and 14cultivars of V. vinifera L. Also, it has provided a solid basis for molecular marker-assisted selection (MAS)and for possibly cloning disease resistance genes in the future.

  1. Estimation of genetic diversity Among Turkish kale populations (Brassica oleracea var. acephala L.) using RAPD markers.

    Science.gov (United States)

    Okumus, A; Balkaya, A

    2007-04-01

    20 populations of kale (B. oleracea var. acephala L.) selected from 127 populations for fresh consumption terms of yield and leaf quality characteristics as superior types using weight-based ranking method from the Black Sea Region of Turkey were evaluated at the DNA level using randomly amplified polymorphic DNA (RAPD) markers compared to some morphological characters. The 7 primers selected from 100 decamers used generated 110 bands, of which 60 (54.5%) were polymorphic. Jaccard's genetic distances were calculated and dendogram was generated using the UPGMA algorithm. The dendogram obtained were classified into three main groups and four subgroups. The accessions showed a limited clustering in compare to morphological characters such as the number of leaf, leaf intentation of the margin, leaf and midrib color and thickness of midrib than geographical characteristics. Leaf color and midrib thickness characters clustered in the same group as OR49 and G18 accessions; S20, G6 and OR37 accessions, respectively.

  2. RAPD Analysis of M1 Generation of Gladiolus hybridus Hort Treated by EMS

    Institute of Scientific and Technical Information of China (English)

    WANG Jingang; GUO Ying; CHE Daidi; LIU Shenkui; YANG Chuanpin

    2009-01-01

    Gladiolus hybridus Hort is one of the world's famous cutting flowers. It is very popular because of the big size, bright color, various shape, and long period of bloom. New species should be cultivated in order to meet the consumers' need of asking for the new. Among the technologies of cultivating new species of flowers, mutagenic breeding is a shortcut. This study treated corm bud of G. hybridus Hort with EMS of different consistency. Then M1 after treated was analyzed by RAPD. The result showed that EMS was a very effective mutagenic agent for the corm bud of G. hybridus Hort. With the increase of consistency, the mutagenic range increased first, then decreased, among which 0.6% EMS treatment had the biggest influence. However, with the same EMS consistency, there was not close relevancy between the amount of mutagenic agent and the divergence of plant's genomes, which offered a molecular basis for selecting plants with good mutation.

  3. Inferences on the role of insertion in a mutation accumulation experiment with Drosophila melanogaster using RAPDs.

    Science.gov (United States)

    Salgado, C; Nieto, B; Toro, M A; López-Fanjul, C; García-Dorado, A

    2005-01-01

    The genetic variability for RAPDs band pattern was studied in a set of 157 mutation accumulation (MA) lines of Drosophila melanogaster. These MA lines were derived from the same isogenic base population and subsequently maintained by full-sib mating during 132 generations. The ancestral pattern of the original isogenic base can be unambiguously established as the consensus pattern of the MA lines and, because these lines are expected to be homozygous, dominance for band pattern is not a concern. Only repeatable changes in band pattern were considered. The number of ancestral bands detected implies that nine-nucleotide targets are enough for repeatable PCR amplification. Compared with the ancestral pattern, one MA line lost one band and two MA lines gained a new one. These results can be accounted for by the insertion of transposable elements occurring at a rate 0.07 Idefix family is also active in the lines.

  4. Genetic variability in wild genotypes of Passiflora cincinnata based on RAPD markers.

    Science.gov (United States)

    Cerqueira-Silva, C B M; Conceição, L D H C S; Santos, E S L; Cardoso-Silva, C B; Pereira, A S; Oliveira, A C; Corrêa, R X

    2010-12-21

    The genetic diversity and characteristics of commercial interest of Passiflora species make it useful to characterize wild germplasm, because of their potential use for fruit, ornamental and medicinal purposes. We evaluated genetic diversity, using RAPD markers, of 32 genotypes of Passiflora cincinnata collected from the wild in the region of Vitória da Conquista, Bahia, Brazil. Thirteen primers generated 95 polymorphic markers and only one monomorphic marker. The mean genetic distance between the genotypes estimated by the complement of the Dice index was 0.51 (ranging from 0.20-0.85), and genotype grouping based on the UPGMA algorithm showed wide variability among the genotypes. This type of information contributes to identification and conservation of the biodiversity of this species and for the identification of pairs of divergent individuals for maximum exploitation of existing variability.

  5. Molecular identification of sex in Simarouba glauca by RAPD markers for crop improvement strategies

    Directory of Open Access Journals (Sweden)

    Gayatri Vaidya

    2014-12-01

    Full Text Available Due to lack of morphological methods to identify sex at early stage in the plants with long juvenile period the application of molecular markers is expected to facilitate breeding program. The objective of this study is to identify molecular markers linked to sex determination of the plant Simarouba glauca which assists in crop improvement program. Random amplified polymorphic DNA primers were tested on dioeceious and hermaphrodite plant Simarouba glauca. A set of eighty five RAPD primers were screened out of which only five primers were found to be associated with sex. The primer OPU-10 is male specific and OPD-19 primer is female specific. Another primer OPU-19 produced a unique amplification in only hermaphrodite individuals. Female and hermaphrodite specific primer OPS-05 amplified an amplicon in female and hermaphrodite and was absent in male plant. Primer OPW-03 produced amplicon specific to male and hermaphrodite plants and was absent in female plants.

  6. RAPD identification of Varroa destructor genotypes in Brazil and other regions of the Americas.

    Science.gov (United States)

    Guerra, J C V; Issa, M R C; Carneiro, F E; Strapazzon, R; Moretto, G

    2010-01-01

    The mite Varroa destructor is the main pest causing damage to apiculture worldwide. In Brazil and other parts of the world, where bees of African origin and their hybrids predominate, the bees can survive these mites without treatment. Studies have shown a correlation between the various genotypes of the mite and its fertility in different geographical regions. Information about mite genotype could be helpful in understanding the diverse effects and relationships of the mite with bees in different regions of the world. DNA analysis by RAPD technique has permitted identification of three distinct genotypes in the mite V. destructor, namely Russian, Japanese and Papua New Guinea. We found predominance of the Russian genotype in Brazil, along with other parts of South America, and in Cuba and Mexico. The Japanese genotype was exclusively found on Fernando de Noronha Island in Brazil.

  7. Analysis of the genetic diversity of physic nut, Jatropha curcas L. accessions using RAPD markers.

    Science.gov (United States)

    Rafii, M Y; Shabanimofrad, M; Puteri Edaroyati, M W; Latif, M A

    2012-06-01

    A sum of 48 accessions of physic nut, Jatropha curcas L. were analyzed to determine the genetic diversity and association between geographical origin using RAPD-PCR markers. Eight primers generated a total of 92 fragments with an average of 11.5 amplicons per primer. Polymorphism percentages of J. curcas accessions for Selangor, Kelantan, and Terengganu states were 80.4, 50.0, and 58.7%, respectively, with an average of 63.04%. Jaccard's genetic similarity co-efficient indicated the high level of genetic variation among the accessions which ranged between 0.06 and 0.81. According to UPGMA dendrogram, 48 J. curcas accessions were grouped into four major clusters at coefficient level 0.3 and accessions from same and near states or regions were found to be grouped together according to their geographical origin. Coefficient of genetic differentiation (G(st)) value of J. curcas revealed that it is an outcrossing species.

  8. Identification of Intergeneric Hybrid Plants Between Oryza sativa and O. minuta via GISH and RAPD

    Institute of Scientific and Technical Information of China (English)

    YU Shun-wu; CHEN Bao-tang; TAO Ai-lin; ZHANG Duan-pin

    2003-01-01

    To transfer desirable resistance traits from O. minuta to O. sativa, intergeneric hybrid plants between O. sativa (AA, 2n=2X=24) and O. minuta (BBCC, 2n=4X=48) were produced by embryo rescue after sexual cross. Morphological observation and chromosome counts indicated their hybrid status (ABC, 2n =3X=36). Genomic in situ hybridization (GISH) was further applied to confirm the parentage of the chromosomes of F1 hybrids. Chromosomes of O. minuta and O. sativa were distinguishable in the hybrids in different fluorescence colors. GISH indicated that A and BC chromosomes were not randomly assembled in a cell.RAPD profiles unequivocally revealed their hybrids with double parent patterns. The results of blast tests showed that the hybrids had obtained disease resistance from O. minuta, and had a level of susceptibility between the parents.

  9. Effect of population size on genetic variation levels in Capparis spinosa (Capparaceae detected by RAPDs

    Directory of Open Access Journals (Sweden)

    Houshang Nosrati

    2012-07-01

    Full Text Available Background: The population size of plants affects on population genetic variation. Materials and Methods: We studied the impact of population size on genetic variation in populations of Capparis spinosa (caper, Capparaceae using RAPDs in East Azerbaijan (Iran. Within-population genetic diversity was estimated based on Nei`s and Shanonn`s diversity using Popgen, and genetic similarity among the populations was studied from a UPGMA dendrogram based the matrix of Nei’s distances obtained through SHAN. Difference in the level genetic variation between small-sized and large-sized populations was tested using Mann-Whitney U test, and correlation between geographical and genetic distances among populations was examined by Pearson test (SPSS, 11.3. Total genetic variation was partitioned into within and among populations based on AMOVA using Arlequin. Results: The polymorphism levels of RAPDs bands among the populations ranged from 48.8% to 81.4%, and within-population Nei’s diversity varied from 0.1667 to 0.2630. Genetic variation in small-sized populations (0.1667 to 0.1809 was significantly lower than the variations in large-sized populations (0.2158 -0.2630 (N= 7, P0.674, Pearson correlation test. Conclusions: Population size has a dramatic impact on its genetic diversity. The results revealed that fragmentation of caper population in the study region has most likely occurred recently. The low genetic diversity revealed within caper populations indicates high risk of extinction and suggests that urgent conservation action is needed to recover diversity in these populations.

  10. Assessment of genetic stability in micropropagules of Jatropha curcas genotypes by RAPD and AFLP analysis

    KAUST Repository

    Sharma, Sweta K.

    2011-07-01

    Jatropha curcas (Euphorbiaceae), a drought resistant non edible oil yielding plant, has acquired significant importance as an alternative renewable energy source. Low and inconsistent yields found in field plantations prompted for identification of high yielding clones and their large scale multiplication by vegetative propagation to obtain true to type plants. In the current investigation plantlets of J. curcas generated by axillary bud proliferation (micropropagation) using nodal segments obtained from selected high yielding genotypes were assessed for their genetic stability using Randomly Amplified Polymorphic DNA (RAPD) and Amplified Fragment Length Polymorphism (AFLP) analyses. For RAPD analysis, 21 out of 52 arbitrary decamer primers screened gave clear reproducible bands. In the micropropagated plantlets obtained from the 2nd sub-culture, 4 out of a total of 177 bands scored were polymorphic, but in the 8th and 16th sub-cultures (culture cycle) no polymorphisms were detected. AFLP analysis revealed 0.63%, 0% and 0% polymorphism in the 2nd, 8th and 16th generations, respectively. When different genotypes, viz. IC 56557 16, IC 56557 34 and IC 56557 13, were assessed by AFLP, 0%, 0.31% and 0.47% polymorphisms were found, respectively, indicating a difference in genetic stability among the different genotypes. To the best of our knowledge this is the first report on assessment of genetic stability of micropropagated plantlets in J. curcas and suggests that axillary shoot proliferation can safely be used as an efficient micropropagation method for mass propagation of J. curcas. © 2011 Elsevier B.V.

  11. Estimation of the Genetic Diversity in Tetraploid Alfalfa Populations Based on RAPD Markers for Breeding Purposes

    Directory of Open Access Journals (Sweden)

    Slobodan Katic

    2011-08-01

    Full Text Available Alfalfa is an autotetraploid, allogamous and heterozygous forage legume, whose varieties are synthetic populations. Due to the complex nature of the species, information about genetic diversity of germplasm used in any alfalfa breeding program is most beneficial. The genetic diversity of five alfalfa varieties, involved in progeny tests at Institute of Field and Vegetable Crops, was characterized based on RAPD markers. A total of 60 primers were screened, out of which 17 were selected for the analysis of genetic diversity. A total of 156 polymorphic bands were generated, with 10.6 bands per primer. Number and percentage of polymorphic loci, effective number of alleles, expected heterozygosity and Shannon’s information index were used to estimate genetic variation. Variety Zuzana had the highest values for all tested parameters, exhibiting the highest level of variation, whereas variety RSI 20 exhibited the lowest. Analysis of molecular variance (AMOVA showed that 88.39% of the total genetic variation was attributed to intra-varietal variance. The cluster analysis for individual samples and varieties revealed differences in their population structures: variety Zuzana showed a very high level of genetic variation, Banat and Ghareh were divided in subpopulations, while Pecy and RSI 20 were relatively uniform. Ways of exploiting the investigated germplasm in the breeding programs are suggested in this paper, depending on their population structure and diversity. The RAPD analysis shows potential to be applied in analysis of parental populations in semi-hybrid alfalfa breeding program in both, development of new homogenous germplasm, and identification of promising, complementary germplasm.

  12. Genetic Diversity Caused by Environmental Stress in Natural Populations of Niupidujuan as Revealed by RAPD Technique

    Institute of Scientific and Technical Information of China (English)

    DU Ying-da; XING Ming; YANG Zhi-yong; LIU Yan-fei; CHEN Xia

    2011-01-01

    Multiplex environmental factors are generally expected to have significant effects on genetic diversity of plant populations.In this study,randomly amplified polymorphic DNA(RAPD) technique was used to reveal the genetic diversity in the same species of four populations collected from Niupidujuan(Rhododendron chrysanthum) at different altitudes,an endangered species,endemic to Northeast China.Initially,twenty informative and reproducible primers were chosen for final RAPD analysis.A total of 152 clear bands were obtained,including 143 polymorphic ones.With the help of POPGENE software,the poly rate was calculated to be 94.07% and the evenness of amplified bands for every primer was 6.8.Additionally,the mean observed number of alleles was 1.7265 with an effective number of 1.3608.An examination of the gene indicated a diversity of 0.2162 with an information diversity index of 0.3313.For these data,the clustering blurred analysis was performed with the aid of NTSYS-pc software to define the Nei's gene diversity and the Shannon information diversity index of the four plant populations.The relationships between the genetic diversity indexes on the one hand and the geographic and climatic factors on the other hand were estimated by the Pearson correlation with SPSS 11.0 software.The results of the correlation analysis show that there were significant(P<0.05) or highly significant(P<0.01) correlations between each of the genetic diversity indexes and the different temperature which were mainly caused by the altitude different populations located.These data highlight the importance of native populations in shaping the spatial genetic structure in Niupidujuan.

  13. [Phylogenetic relationships and intraspecific variation of D-genome Aegilops L. as revealed by RAPD analysis].

    Science.gov (United States)

    Goriunova, S V; Kochieva, E Z; Chikida, N N; Pukhal'skiĭ, V A

    2004-05-01

    RAPD analysis was carried out to study the genetic variation and phylogenetic relationships of polyploid Aegilops species, which contain the D genome as a component of the alloploid genome, and diploid Aegilops tauschii, which is a putative donor of the D genome for common wheat. In total, 74 accessions of six D-genome Aegilops species were examined. The highest intraspecific variation (0.03-0.21) was observed for Ae. tauschii. Intraspecific distances between accessions ranged 0.007-0.067 in Ae. cylindrica, 0.017-0.047 in Ae. vavilovii, and 0.00-0.053 in Ae. juvenalis. Likewise, Ae. ventricosa and Ae. crassa showed low intraspecific polymorphism. The among-accession difference in alloploid Ae. ventricosa (genome DvNv) was similar to that of one parental species, Ae. uniaristata (N), and substantially lower than in the other parent, Ae. tauschii (D). The among-accession difference in Ae. cylindrica (CcDc) was considerably lower than in either parent, Ae. tauschii (D) or Ae. caudata (C). With the exception of Ae. cylindrica, all D-genome species--Ae. tauschii (D), Ae. ventricosa (DvNv), Ae. crassa (XcrDcrl and XcrDcrlDcr2), Ae. juvenalis (XjDjUj), and Ae. vavilovii (XvaDvaSva)--formed a single polymorphic cluster, which was distinct from clusters of other species. The only exception, Ae. cylindrica, did not group with the other D-genome species, but clustered with Ae. caudata (C), a donor of the C genome. The cluster of these two species was clearly distinct from the cluster of the other D-genome species and close to a cluster of Ae. umbellulata (genome U) and Ae. ovata (genome UgMg). Thus, RAPD analysis for the first time was used to estimate and to compare the interpopulation polymorphism and to establish the phylogenetic relationships of all diploid and alloploid D-genome Aegilops species.

  14. Micropropagation and assessment of genetic fidelity of Dendrocalamus strictus (Roxb.) nees using RAPD and ISSR markers.

    Science.gov (United States)

    Goyal, Arvind Kumar; Pradhan, Sushen; Basistha, Bharat Chandra; Sen, Arnab

    2015-08-01

    Dendrocalamus strictus popularly known as 'Male bamboo' is a multipurpose bamboo which is extensively utilized in pharmaceutical, paper, agricultural and other industrial implements. In this study, in vitro regeneration of D. strictus through nodal culture has been attempted. Murashige and Skoog's medium supplemented with 4 mg/l BAP was found to be most effective in shoot regeneration with 3.68 ± 0.37 shoots per explant. The effect of Kn was found to be moderate. These hormones also had considerable effect on the shoot length. The highest shoot length after 6 weeks (3.11 ± 0.41 cm) was noted with 5 mg/l BAP followed by 3.07 ± 0.28 cm with 5 mg/l Kn, while decrease in the shoot length was noted with other treatments. The effect of IBA and NAA individually or in combination at different concentrations on rooting was evaluated. The highest number of root (1.36 ± 0.04) was regenerated on full-strength MS medium supplemented with 3 mg/l NAA, while maximum length of 1.64 ± 0.03 cm of roots was recorded with combination of 1 mg/l IBA and 3 mg/l NAA. Tissue-cultured plants thus obtained were successfully transferred to the soil. The clonal fidelity among the in vitro-regenerated plantlets was assessed by RAPD and ISSR markers. The ten RAPD decamers produced 58 amplicons, while nine ISSR primers generated a total of 66 bands. All the bands generated were monomorphic. These results confirmed the clonal fidelity of the tissue culture-raised D. strictus plantlets and corroborated the fact that nodal culture is perhaps the safest mode for multiplication of true to type plants.

  15. RAPD and mitochondrial DNA analysis of the soybean stalk weevil, Sternechus subsignatus (Coleoptera: Curculionidae).

    Science.gov (United States)

    Sosa-Gomez, D R; Coronel, N; Binneck, E; Zucchi, M I; Rosado-Neto, G

    2008-10-01

    Sternechus subsignatus Boheman (Curculionidae: Sternechini) is one of the primary Curculionidae species that reduces soybean yield in Brazil. Initially, outbreaks were reported in southern Brazil in 1973; but, more recent, outbreaks were reported in Bahia (summer 1997-1998) and Maranhão (summer 2003-2004), two states in northeastern Brazil. A putative related species, S. pinguis (Fabricius), was first detected in Salta Province, Argentina. The objective of this study was to evaluate intraspecific molecular polymorphisms of geographically distinct Sternechus populations. Randomly amplified polymorphic DNA (RAPD) profiles and partial mitochondrial cytochrome B (CytB) gene sequences were used to determine whether individual soybean stalk weevils were one of two different species and to infer pest invasion pattern. Putative S. pinguis and S. subsignatus populations were collected in San Agustin (Cruz Alta, Tucumán Province, Argentina) and different sampling sites in the Brazilian states of Paraná, Bahia and Maranhão. Polymorphic bands were obtained by RAPD and analyzed by Dice coefficients. Populations from southern Brazil were more closely related genetically to an Argentinean group than the populations sampled in northeastern Brazil. The Londrina Co., Brazil population displayed the highest intra-population genetic similarity. Most of the soybean stalk weevils collected from San Agustin, Tucumán, Argentina were divergent from those collected in Brazil. Sequencing and parsimony analysis of CytB did not differentiate specimens collected in Argentina and Brazil. Thus, our data show that soybean stalk weevil outbreaks and population increases in northeastern Brazil involved local genotypes.

  16. Eficacia de un Enjuagatorio Comercial en Comparación al Enjuagatorio con Cloruro de Sodio al 5% en la Disminución de Streptococcus Mutans

    OpenAIRE

    Cayo, Cesar; Universidad Alas Peruanas; Salvador, Sandra; Ramos, Javier

    2015-01-01

    El presente trabajo tiene como finalidad determinar el nivel de eficacia del enjuagatorio comercial en comparación con el enjuagatorio con cloruro de sodio al 5% en la disminución del Streptococcus mutans en la placa dentobacteriana. El grupo experimental evaluado estuvo constituido de 45 alumnos del 2¡Æ de secundaria del Centro Educativo.Felix B. Cardenas. Los participantes se agruparon al azar, de la siguiente manera: Grupo n¡Æ01 uso el Enjuagatorio comercial, Grupo n¡Æ02 usaron Cloruro de ...

  17. Variabilidade genética em búfalos estimada por marcadores RAPD Genetic variability of buffaloes estimated by RAPD markers

    Directory of Open Access Journals (Sweden)

    Maria do Socorro Maués Albuquerque

    2006-04-01

    Full Text Available O objetivo deste trabalho foi caracterizar, por meio de marcadores RAPD, dois grupos genéticos de búfalos, Carabao e tipo Baio, que estão sendo conservados in situ, assim como verificar as relações genéticas entre eles e os outros três grupos genéticos de búfalos existentes no Brasil, Murrah, Jafarabadi e Mediterrâneo, considerados raças comerciais. Foram estudados 48 animais de cada grupo, com exceção dos grupos Murrah e Mediterrâneo, com 47 e 42 animais, respectivamente, compreendendo um total de 233 animais. Os 21 iniciadores polimórficos geraram 98 marcadores. A variabilidade genética entre e dentro dos grupos foi estimada em 26,5 e 73,5%, respectivamente, sugerindo divergência significativa entre os cinco grupos genéticos. Na análise entre pares de grupos, foi verificado que a maior e a menor divergência estavam em torno de 40 e 18%, quando se compararam os grupos Carabao x Mediterrâneo e Murrah x Jafarabadi, respectivamente. Entre os grupos Baio e Murrah, a análise revelou divergência genética de 20,42%, indicando que esses grupos são distintos. Os cinco grupos são geneticamente distintos, o que reforça a necessidade de conservação dos grupos genéticos Carabao e Baio, ameaçados de extinção no Brasil.The objective of this work was to characterize genetically, using RAPD markers, two genetic groups of buffalos, Carabao and Baio, which are being conserved in situ, as well as to verify the genetic relationship among them and the other three genetic groups of buffalos raised in Brazil, considered as commercial breeds: Murrah, Jaffarabadi and Mediterrâneo. Forty eight animals of each group were studied, with the exception of the Murrah and Mediterrâneo, in which 47 and 42 animals, respectively, were sampled, comprising a total of 233 animals. The 21 polymorphic primers produced 98 markers. Genetic variability within and between groups was estimated in 26.5 and 73.5%, respectively, suggesting a significant

  18. Divergência genética entre cultivares de gérbera utilizando marcadores RAPD Genetic divergence among cultivars of gerbera using RAPD markers

    Directory of Open Access Journals (Sweden)

    Rodrigo Kelson Silva Rezende

    2009-11-01

    Full Text Available No processo de produção comercial de mudas de gérbera, a cor da flor é uma das principais características morfológicas de interesse agronômico, sendo uma característica importante em programas de melhoramento genético. A utilização de marcadores moleculares pode servir para direcionar cruzamentos, confirmar novos híbridos ou genótipos mutantes e identificar novos genótipos para fins comerciais. Nesse contexto, o objetivo deste trabalho foi analisar a divergência genética entre seis cultivares de Gerbera jamesonii ('Jaguar Yellow', 'Jaguar Cream', 'Jaguar Lemon', 'Jaguar Salmon Pastel', 'Jaguar Red', 'Jaguar Deep Rose'. A análise de divergência genética entre as cultivares de gérbera foi realizada utilizando-se 21 primers, os quais amplificaram 37 fragmentos polimórficos de DNA, que foram usados para estimar o coeficiente de Jaccard, o qual apresentou uma média de 0,38, variando de 0,28 a 0,56. A estrutura genética entre as cultivares foi estimada pelo UPGMA, revelando dois grupos distintos, a 38% de similaridade genética. A maior similaridade genética encontrada (56% foi entre as cultivares 'Jaguar Yellow' e 'Jaguar Lemon'. Os resultados demonstram que a técnica RAPD oferece uma maneira rápida, relativamente barata e útil para a caracterização da divergência genética entre as diferentes cultivares de Gerbera jamesonii com relação à cor da flor.During the commercial production of gerbera seedlings, flower color is one of the main morphological aspects that have an agronomic interest and becoming an important feature in genetic breeding programs. The use of molecular markers may serve to direct crossings, new hybrids and mutants, besides confirm and identify new genotypes for commercial purposes. In that context, this work aimed to analyze the genetic divergence among six cultivars of Gerbera jamesonii ('Jaguar Yellow', 'Jaguar Cream', 'Jaguar Lemon', 'Jaguar Salmon Pastel', 'Jaguar Red', 'Jaguar Deep Rose'. The

  19. Random amplification of polymorphic DNA (RAPD), pulsed-field gel electrophoresis (PFGE) and phage-typing in the analysis of a hospital outbreak of Salmonella enteritidis

    DEFF Research Database (Denmark)

    Skibsted, U.; Baggesen, Dorte Lau; Dessau, R.

    1998-01-01

    Isolates of Salmonella Enteritidis from 81 patients from Herlev Hospital or from Copenhagen County were analysed by random amplification of polymorphic DNA (RAPD), pulsed-field gel electrophoresis (PFGE) and phage-typing. Fourteen polymorphic markers from five decamer primers unambiguously placed...... that RAPD is useful as a tool in investigations of microbial outbreaks in its own right, or to supplement phage-typing and PFGE of Salmonella Enteritidis....

  20. Molecular analysis of RAPD DNA based markers: their potential use for the detection of genetic variability in jojoba (Simmondsia chinensis L Schneider).

    Science.gov (United States)

    Amarger, V; Mercier, L

    1995-01-01

    We have applied the recently developed technique of random amplified polymorphic DNA (RAPD) for the discrimination between two jojoba clones at the genomic level. Among a set of 30 primers tested, a simple reproducible pattern with three distinct fragments for clone D and two distinct fragments for clone E was obtained with primer OPB08. Since RAPD products are the results of arbitrarily priming events and because a given primer can amplify a number of non-homologous sequences, we wondered whether or not RAPD bands, even those of similar size, were derived from different loci in the two clones. To answer this question, two complementary approaches were used: i) cloning and sequencing of the amplification products from clone E; and ii) complementary Southern analysis of RAPD gels using cloned or amplified fragments (directly recovered from agarose gels) as RFLP probes. The data reported here show that the RAPD reaction generates multiple amplified fragments. Some fragments, although resolved as a single band on agarose gels, contain different DNA species of the same size. Furthermore, it appears that the cloned RAPD products of known sequence that do not target repetitive DNA can be used as hybridization probes in RFLP to detect a polymorphism among individuals.

  1. Estimación mediante RAPD's de la diversidad genética en Guadua en el departamento del Cauca, Colombia

    Directory of Open Access Journals (Sweden)

    Palacio M. Juan Diego

    2006-06-01

    Full Text Available

    Mediante RAPD's se analizaron 120 muestras foliares de 12 biotipos de Guadua angustifolia Kunth clasificados morfológicamente, procedentes de la cuenca del río Cauca, en el departamento del Cauca, Colombia, para determinar diversidad genética. El ADN se extrajo mediante el protocolo modificado de Dellaporta (1983. Se emplearon los cebadores; OPF-12, OPG-19, OPN-19 y OPP-16 con mayor número de bandas polimórficas. El índice de Shannon (HT = 0.4556 ± 0.1849 señaló diversidad genética total alta y diversidad entre los biotipos y al interior de ellos. El Índice de estructura genética (Gst = 0.5200 e Indice de migración efectiva (Nm = 0.4615 definieron biotipos bien diferenciados. El análisis de similaridad conformó tres grupos a un coeficiente de 0.64. El grupo G1 incluyó los biotipos Curvado, Rayada frecuente, Amarilla Playón, Rayada ancha, Rayada escasa, Convexa, Amarilla, Hembra, Verde irregular y algunos individuos de verde alta. El grupo G2, Verde alta y Macho. El grupo G3, Rayada negra. El estudio molecular agrupó los individuos de forma similar al estudio morfológico, con excepción de los individuos del biotipo Hembra.

    Palabras claves: Guadua angustifolia, caracterización molecular, variación genética.

  2. Polyembryony and identification of Volkamerian lemon zygotic and nucellar seedlings using RAPD Poliembrionia e identificação de seedlings zigóticos e nucelares de limão volkameriano com RAPD

    Directory of Open Access Journals (Sweden)

    María Andrade-Rodríguez

    2004-06-01

    Full Text Available The objectives of this work were to evaluate the frequency of polyembryony, and to identify zygotic and nucellar seedlings of Citrus volkameriana using RAPD. Twenty-five polyembryonic and eight monoembryonic seeds were cultivated in vitrofor six months. DNA from seedlings was extracted and used in combination with five RAPD primers to identify zygotic or nucellar origin of the seedlings. Environmental conditions of the year affected significantly (POs objetivos deste trabalho foram avaliar a freqüência da poliembrionia e indentificar os seedlings zigóticos e nucelares de Citrus volkameriana usando RAPD. Cultivaram-se in vitro 25 sementes poliembriônicas e 8 monoembriônicas, por seis meses. Extraiu-se o DNA dos seedlings e usou-se em combinação com cinco primers RAPD para identificar a origem zigótica e nucelar das plântulas. Os resultados mostraram que as condições ambientais do ano afetaram significativamente (P<0,05 as características morfológicas dos frutos e o número de embriões por semente. Os índices de sementes poliembriônicas variaram nos três anos de avaliação (30,9%, 44,8% e 54,4%. As características morfológicas não se correlacionaram com a poliembrionia. A cultura in vitro possibilita que todos os embriões de cada semente cresçam, favorecendo os seedlings identificados como zigóticos. Nas sementes poliembriônicas e monoembriônicas, 25,9% e 87,5% dos seedlings, respectivamente, originaram-se sexualmente. Nem todos os seedlings zigóticos foram produzidos por embriões localizados no micrófilo das sementes poliembriônicas.

  3. Mapping of risk prone areas of kala-azar (Visceral leishmaniasis in parts of Bihar state, India: an RS and GIS approach

    Directory of Open Access Journals (Sweden)

    S. Sudhakar, T. Srinivas, A. Palit, S.K. Karc, S.K. Battacharya

    2006-09-01

    Full Text Available Background & objectives: The kala-azar fever (Visceral leishmaniasis is continuing unabated inIndia for over a century, now being largely confined to the eastern part of India mainly in Bihar stateand to some extent in its bordering states like West Bengal and Uttar Pradesh. Two study sites namelyPatepur block in Vaishali district with high endemicity in northern part and Lohardagga block inLohardagga district with absolute non-endemicity in southern part of Bihar were selected for thestudy with the following objectives : (i to study the macro-ecosystem in relation to distribution ofvector —Phlebotomus argentipes; (ii to identify/map the risk prone areas or villages in a block forquick remedial measures; and (iii to make use of satellite remote sensing and GIS to demonstratethe utility for rapid assessment of landuse/landcover and their relation with the incidence of kalaazarleading to the mapping of risk prone areas.Methods: Indian Remote Sensing (IRS-1D LISS III satellite data for the periods of March andNovember 2000 were analysed in Silicon graphic image processing system using ERDAS software.False color composites (FCC were generated and landuse/landcover was assessed using Maximumlikelihood supervised classification techniques based on ground truth training sets. During the studythe GIS functions are used to quantify the remotely sensed landscape proportions of 5 km2 buffersurrounding each known group of villages of high occurrence of sandflies in endemic and nonendemicstudy sites. Instead of traditional ground based survey methods to vector surveillance, thepresent study used a combination of remote sensing (RS and geographical information system (GISapproach to develop landscape predictors of sandfly abundance—an indicator of human vector contactand as a measure of risk prone areas.Results: Statistical analysis using the remotely sensed landscape variables showed that rural villagessurrounded by higher proportion of transitional

  4. Optimization of RAPD-PCR Reaction System for Onion (Allium cepa L. )by Orthogonal Design%洋葱(Allium cepa L.)RAPD-PCR反应体系及扩增程序的优化

    Institute of Scientific and Technical Information of China (English)

    陈沁滨; 侯喜林; 王建军; 冷月强; 蒋芳玲; 薛萍

    2006-01-01

    为建立多态性高、稳定性好的洋葱RAPD-PCR反应体系,采用正交设计,研究了Taq酶、Mg2+、引物和dNTP 4种RAPD-PCR反应组分浓度变化对扩增结果的影响,在此基础上对模板DNA用量、扩增程序中退火温度和反应循环次数进行了筛选.试验结果表明,洋葱20 μl RAPD-PCR优化反应体系为1×Buffer、2.0 mmol/L Mg2+、1.0 U Taq DNA聚合酶、200 μmol/L dNTP、0.6 μmol/L引物、2%甘油和15 ng DNA模板;PCR扩增程序为94℃预变性4 min;94℃变性30 s,35℃退火40 s,72℃延长1.5 min,45个循环;72℃保温延伸7 min.

  5. Application of RAPD Markers in Laboratory Animal%RAPD技术在实验动物中的应用

    Institute of Scientific and Technical Information of China (English)

    张建珍; 马恩波

    2003-01-01

    With the sharp development of biomedicine, the protection and utilization of laboratory animal resources, and standardization of laboratory animal have been regarded as the theme of laboratory animal science. Many kinds of genetic markers were used in genetic background analysis, identification of species and strains, and breeding of laboratory animal, unlike morphological marker, cytological marker and isozyme marker, which revealed the difference at gene expressing level, molecular genetic markers used as an accurate and effective method reveal genetic variations of hereditary material, it plays an important role in many fields. The RAPD (Random Amplification Polymorphic DNA) method was firstly described in 1990, it has several distinct advantages over other techniques in that it is easy to perform and fast, and so was widely applied to taxonomy and identification of species, construction of genetic mapping, genetic relationships among species and population genetics. This paper summarized the application of RAPD markers in laboratory animal in recent 10 years.

  6. Genomic characterization of lung flukes, Paragonimus heterotremus, P. siamensis, P. harinasutai, P. westermani and P. bangkokensis by RAPD markers.

    Science.gov (United States)

    Intapan, P M; Kosuwan, T; Wongkham, C; Maleewong, W

    2004-09-20

    Random amplified polymorphic DNA (RAPD) markers were assayed in an attempt to discriminate among five species of Paragonimus. Genomic DNAs of two strains of Paragonimus heterotremus from two provinces in Thailand, Saraburi and Phitsanulok, as well as of P. siamensis, P. harinasutai, P. westermani and P. bangkokensis were extracted and amplified by an arbitrary primer, namely P2 (5-GTTTCGCTCC-3). RAPD patterns showed that those five species were genetically distinct, although they shared genomic DNA to some extent. This primer could also distinguish between two strains of P. heterotremus. The polymorphism observed allowed to construct a relationship dendrogram. The phylogenetic dendrogram showed that the P. heterotremus strains were closest to P. harinasutai, followed by P. siamensis, P. bangkokensis and P. westermani.

  7. Genetic Diversity and Evolutionary Tendency Detected by Isozyme, RFLP and RAPD Markers in the Wild and Cultivated Soybean in China

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    A large numbers of samples of wild soybean accessions and cultivated soybean landraces from various areas in China were analyzed by isozyrme, cytoplasmic DNA RFLP and nuclear DNA RAPD markers in order to reveal their genetic diversity. Greater comprehensive genetic diversity was detected in wild soybean than in cultivated soybean. The genetic plentifulness and the genetic dispersion of wild soybean were 180 (95. 2%) and 0. 2891 while those of cultivated soybean were 154(81.5%) and 0. 2091,respectively. On the most loci, especially on isozyme loci Idh1, Aph, Idh2,and Dia, cytoplasmic DNA RFLP loci cp Ⅰ , cp Ⅲ, mt Ⅳ a and mt Ⅳ b, and nuclear RAPD loci OPAP4-8, OPAP5-1, OPAP9-8 and OPAP20-8, the wild soybeans djffered remarkably from the cultivated ones in allele frequency. These markers could be used in further study on the evolution and origin of the cultivated soybean.

  8. Genetic Diversity and Evolutionary Tendency Detected by Isozyme, RFLP and RAPD Markers in the Wild and Cultivated Soybean in China

    Institute of Scientific and Technical Information of China (English)

    Xu Donghe; Gao Zhong; Gai Junyi; Zhang Zhiyong; Chen Shouyi; Fukushi Hirofurm; Kitajirma Shunji; Abe Jun; Shimamoto Yoshiya

    2000-01-01

    A large numbers of samples of wild soybean accessions and cultivated soybean landraces from various areas in China were analyzed by isozyrme, cytoplasmic DNA RFLP and nuclear DNA RAPD markers in order to reveal their genetic diversity. Greater comprehensive genetic diversity was detected in wild soybean than in cultivated soybean. The genetic plentifulness and the genetic dispersion of wild soybean were 180 (95. 2%) and 0. 2891 while those of cultivated soybean were 154(81.5%) and 0. 2091,respectively. On the most loci, especially on isozyme loci Idh1, Aph, Idh2,and Dia, cytoplasmic DNA RFLP loci cp Ⅰ , cp Ⅲ, mt Ⅳ a and mt Ⅳ b, and nuclear RAPD loci OPAP4-8, OPAP5-1, OPAP9-8 and OPAP20-8, the wild soybeans djffered remarkably from the cultivated ones in allele frequency. These markers could be used in further study on the evolution and origin of the cultivated soybean.

  9. Detection of DNA Hypomethylation Mediated Floral Induction in Longan and Spinach Using the HAT-RAPD Technique

    Directory of Open Access Journals (Sweden)

    S. Anuntalabhochai

    2009-01-01

    Full Text Available In this study, the HAT-RAPD technique was used to detect DNA methylation in the four plant species, rice (Oryza sativa, petunia (Petunia hybrida, spinach (Spinacea oleracea L. and longan (Dimocarpus longan Lour., which were treated using 5-azacytidine, potassium chlorate (KClO3 and a low temperature induction. Rice and petunia were chosen to be induced because in these species it is known that 5-azacytidine is able to induce hypomethylation in their genomes leading to morphological changes, in particular floral induction, in the developing plants. Using the HAT-RAPD technique, DNA methylation was detected using the restriction enzymes HpaII and MspI in rice and petunia (as expected and in spinach, but was found to be absent in longan, which suggests that floral induction in longan is undetectable in longan.

  10. Identification of Co-Segregating RAPD Marker Linked to Powdery Mildew Resistance Gene Pm 18 in Wheat

    Institute of Scientific and Technical Information of China (English)

    ZHANG Qing-li; GU Feng; LI Tao; GAO Ju-rong; WANG Hong-gang

    2004-01-01

    The Pm18 gene of wheat confers resistance to the powdery mildew which is oneof the most serious diseases in many regions of the world. In this study, bulked segregant analysis (BSA) was used to develop randomly amplified polymorphic DNA (RAPD) markers linked to Pml8 gene. Three hundred and twenty decamer primers were screened and one of them was identified as RAPD marker (S411600) linked to Pml8. Using the F2 mapping population from the cross Pml8 × Chancellor, the marker S411600 was shown to co-segregate with the gene Pml8. This marker can be conveniently used for marker-assisted selection in wheat breeding programs for the identification or pyramiding of Pml8 with other resistance genes.

  11. Occurrence and HAT-RAPD analysis of gastrointestinal helminths in domestic chickens (Gallus gallus domesticus) in Phayao province, northern Thailand

    OpenAIRE

    Preeyaporn Butboonchoo; Chalobol Wongsawad

    2015-01-01

    The present study determined the prevalence and distribution of gastrointestinal helminths in domestic chickens (Gallus gallus domesticus) between November 2012 and August 2013. One hundred and twenty domestic chickens were purchased from villages in four districts of Phayao province; Mae Chai, Dok Khamtai, Chun and Chiang Kham. Morphological differences were used to identify the helminth species, and HAT-RAPD technique was used to differentiate among closely related species. The results reve...

  12. Prevalence, antibiotic resistance and RAPD typing of Campylobacter species isolated from ducks, their rearing and processing environments in Penang, Malaysia.

    Science.gov (United States)

    Adzitey, Frederick; Rusul, Gulam; Huda, Nurul; Cogan, Tristan; Corry, Janet

    2012-03-15

    We report for the first time on the prevalence, antibiotic resistance and RAPD types of Campylobacter species in ducks and duck related environmental samples in Malaysia. Samples were examined by enrichment in Bolton Broth followed by plating onto modified Charcoal Cefoperazone Deoxycholate agar (mCCDA) and/or plating directly onto mCCDA. A total of 643 samples were screened, and the prevalence of Campylobacter spp. in samples from different sources ranged from 0% to 85%. The method of isolation had a significant (P<0.05) effect on the isolation rate. One hundred and sixteen Campylobacter isolates, comprising of 94 Campylobacter jejuni, 19 Campylobacter coli and three Campylobacter lari, were examined for their sensitivity to 13 antibiotics. Majority of the C. jejuni isolates were resistant to cephalothin (99%), tetracycline (96%), suphamethoxazole/trimethoprim (96%), and very few were resistant to gentamicin (5%), chloramphenicol (7%) and erythromycin (1%). All C. coli isolates were resistant to cephalothin, nalidixic acid, norfloxacin and tetracycline but susceptible to chloramphenicol, erythromycin and gentamicin. The three C. lari isolates were resistant to all the antibiotics tested except chloramphenicol and gentamicin (1/3 and 2/3 susceptible, respectively). Genetic diversity of Campylobacter isolates were determined using random amplification of polymorphic DNA (RAPD). C. jejuni and C. coli isolates belong to fifty-eight and twelve RAPD types, respectively.

  13. Occurrence and HAT-RAPD analysis of gastrointestinal helminths in domestic chickens (Gallus gallus domesticus) in Phayao province, northern Thailand.

    Science.gov (United States)

    Butboonchoo, Preeyaporn; Wongsawad, Chalobol

    2017-01-01

    The present study determined the prevalence and distribution of gastrointestinal helminths in domestic chickens (Gallus gallus domesticus) between November 2012 and August 2013. One hundred and twenty domestic chickens were purchased from villages in four districts of Phayao province; Mae Chai, Dok Khamtai, Chun and Chiang Kham. Morphological differences were used to identify the helminth species, and HAT-RAPD technique was used to differentiate among closely related species. The results revealed that the total prevalence of infection was 99.2%. Cestode and nematode infections showed the highest prevalence in rainy season, while trematode infections were low and only found in hot season. The species and their prevalence were: Ascaridia galli (50.8%), Heterakis gallinarum (86.7%), Prosthogonimus macrorchis (1.7%), Echinostoma revolutum (0.8%), Raillietina echinobothrida (48.3%), Raillietina tetragona (57.5%), Raillietina cesticillus (12.5%), Raillietina sp. (35.8%), Cotugnia chiangmaii (14.2%) and Cotugnia sp. (32.5%). The prevalence of helminth infections did not differ significantly between male and female chickens. HAT-RAPD analysis, the specific fragment of 400 and 250 bp indicated that Raillietina sp. and Cotugnia sp. found, respectively, differ from other closely related species. This study has confirmed that HAT-RAPD technique can be used to differentiate among related species combined with morphological observations.

  14. Occurrence and HAT-RAPD analysis of gastrointestinal helminths in domestic chickens (Gallus gallus domesticus in Phayao province, northern Thailand

    Directory of Open Access Journals (Sweden)

    Preeyaporn Butboonchoo

    2017-01-01

    Full Text Available The present study determined the prevalence and distribution of gastrointestinal helminths in domestic chickens (Gallus gallus domesticus between November 2012 and August 2013. One hundred and twenty domestic chickens were purchased from villages in four districts of Phayao province; Mae Chai, Dok Khamtai, Chun and Chiang Kham. Morphological differences were used to identify the helminth species, and HAT-RAPD technique was used to differentiate among closely related species. The results revealed that the total prevalence of infection was 99.2%. Cestode and nematode infections showed the highest prevalence in rainy season, while trematode infections were low and only found in hot season. The species and their prevalence were: Ascaridia galli (50.8%, Heterakis gallinarum (86.7%, Prosthogonimus macrorchis (1.7%, Echinostoma revolutum (0.8%, Raillietina echinobothrida (48.3%, Raillietina tetragona (57.5%, Raillietina cesticillus (12.5%, Raillietina sp. (35.8%, Cotugnia chiangmaii (14.2% and Cotugnia sp. (32.5%. The prevalence of helminth infections did not differ significantly between male and female chickens. HAT-RAPD analysis, the specific fragment of 400 and 250 bp indicated that Raillietina sp. and Cotugnia sp. found, respectively, differ from other closely related species. This study has confirmed that HAT-RAPD technique can be used to differentiate among related species combined with morphological observations.

  15. Selection and RAPD analysis of Pseudomonas ssp. isolates able to improve biological viability of potato seed tubers

    Directory of Open Access Journals (Sweden)

    Poštić Dobrivoj

    2013-01-01

    Full Text Available Indoleacetic acid (IAA producing Pseudomonas isolates from the rhizosphere of maize (Q4 and Q20, alfalfa (Q1 and Q16 and wild red clover (B25 were selected for the investigation of their effect on the biological vitality of the potato seed tubers. The production of IAA ranged from 4.09 to 15.9 µ gmL- 1 after 24h of cultivation and 4.08 to 26.4 µ gmL-1 after 48h of cultivation. The molecular comparison by RAPD analysis also was done. RAPD patterns of selected Pseudomonas spp. isolates obtained by BC318, AF14, SPH 1 and AP 10 primers demonstrated the suitability of RAPD method in distinguishing a high variability among the four isolates (44 to 68%. The effect on the biological viability of potato (industrially important variety Pirol was observed during the seven weeks of sprouting at the temperature of 18-20°C. Potato tubers treated by the selected isolates formed slightly lower number of sprouts, but statistically higher mean length - up to 129.9% higher than the control. The mean sprouting capacity was 64.5% higher than a control. Obtained results suggested positive effects of selected IAA producing Pseudomonas isolates on the length of potato tubers and sprouting capacity as the parameters which define biological viability. [Projekat Ministarstva nauke Republike Srbije, br. III46007

  16. Consistency of population genetics parameters estimated from isozyme and RAPDs dataset in species of genus Prosopis (Leguminosae, Mimosoideae).

    Science.gov (United States)

    Ferreyra, Laura Inés; Bessega, Cecilia; Vilardi, Juan C; Saidman, Beatriz O

    2007-11-01

    Genetic variability, population structure and differentiation among 17 populations of 5 species and 2 natural interspecific hybrids of section Algarobia of genus Prosopis were analyzed from data of 23 isozyme and 28 RAPD loci. Both markers indicated that the studied populations are highly variable. P. alba populations in average showed lower values of genetic variability estimates from isozyme data, but this trend was not observed for RAPD markers. The hierarchical analyses of the distribution of genetic variability showed that the highest proportion of variation occurred within populations, the differentiation among species was intermediate and the lowest component was observed among populations within species. The consistency between results from both dataset implies that they are not biased and reflect the actual genetic structure of the populations analyzed. The matrices of Euclidean distances obtained from the two sets of markers were highly correlated according to Mantel test. In both cases the corresponding phenogram and MDS plot tended to cluster conspecific populations while hybrid populations were not intermediate between putative parents. Some disagreements between isozyme and RAPD phenograms were observed mainly in the affinities of hybrid populations. Such inconsistencies might result from reticular rather than dichotomic evolutionary relationships. The phenetic associations retrieved gave no support to the division of the section Algarobia into series.

  17. Application of RFLP-PCR-Based Identification for Sand Fly Surveillance in an Area Endemic for Kala-Azar in Mymensingh, Bangladesh

    Directory of Open Access Journals (Sweden)

    Mohammad Shafiul Alam

    2012-01-01

    Full Text Available Mymensingh is the most endemic district for kala-azar in Bangladesh. Phlebotomus argentipes remains the only known vector although a number of sand fly species are prevalent in this area. Genotyping of sand flies distributed in a VL endemic area was developed by a PCR and restriction-fragment-length polymorphism (RFLP of 18S rRNA gene of sand fly species. Using the RFLP-PCR analysis with AfaI and HinfI restriction enzymes, P. argentipes, P. papatasi, and Sergentomyia species could be identified. Among 1,055 female sand flies successfully analyzed for the species identification individually, 64.4% flies was classified as Sergentomyia species, whereas 35.6% was identified as P. argentipes and no P. papatasi was found. Although infection of Leishmania within the sand flies was individually examined targeting leishmanial minicircle DNA, none of the 1,055 sand flies examined were positive for Leishmania infection. The RFLP-PCR could be useful tools for taxonomic identification and Leishmania infection monitoring in endemic areas of Bangladesh.

  18. El azar de la mujer rubia, de Manuel Vicent: memorias de la transición entre la ficción, la prensa y la historia

    Directory of Open Access Journals (Sweden)

    Sofía Bonino

    2013-12-01

    Full Text Available In the next paper I intend to look the novel written by Manuel Vicent El azar de la mujer rubia. The text is presented to us as a new perspective on the transition, this time structured from a female character from which are drawn the wires of the Spanish history since Franco’s death until the present. However, the reading of the work reveals that the character is in the foreground is not another that Adolfo Suárez and that the material that serves as a starting point is the own journalistic of Manuel Vicent. It would think then this novel as an attempt to save from oblivion those newspaper articles of inestimable value –for having been written at the time that the historical events were unfolding– but also as a work that was presented as novel to reveal the hidden story of a love triangle that directly influenced the history of the country, with a female character that is, for the first time, recovered and processed into heroin. The study will focus on the way in which Vicent retrieves those journalistic texts and in the operations of inclusion of the same in a text that mixes fiction with history to return, once more, to the one of the momentous of the recent Spanish history

  19. Comparison of optical microscopy and quantitative polymerase chain reaction for estimating parasitaemia in patients with kala-azar and modelling infectiousness to the vector Lutzomyia longipalpis

    Science.gov (United States)

    Silva, Jailthon C; Zacarias, Danielle A; Silva, Vladimir C; Rolão, Nuno; Costa, Dorcas L; Costa, Carlos HN

    2016-01-01

    Currently, the only method for identifying infective hosts with Leishmania infantum to the vector Lutzomyia longipalpis is xenodiagnosis. More recently, quantitative polymerase chain reaction (qPCR) has been used to model human reservoir competence by assuming that detection of parasite DNA indicates the presence of viable parasites for infecting vectors. Since this assumption has not been proven, this study aimed to verify this hypothesis. The concentration of amastigotes in the peripheral blood of 30 patients with kala-azar was microscopically verified by leukoconcentration and was compared to qPCR estimates. Parasites were identified in 4.8 mL of peripheral blood from 67% of the patients, at a very low concentration (average 0.3 parasites/mL). However, qPCR showed 93% sensitivity and the estimated parasitaemia was over a thousand times greater, both in blood and plasma, with higher levels in plasma than in blood. Furthermore, the microscopic count of circulating parasites and the qPCR parasitaemia estimates were not mathematically compatible with the published proportions of infected sandflies in xenodiagnostic studies. These findings suggest that qPCR does not measure the concentration of circulating parasites, but rather measures DNA from other sites, and that blood might not be the main source of infection for vectors. PMID:27439033

  20. Phylogeny of certain biocontrol agents with special reference to nematophagous fungi based on RAPd.

    Science.gov (United States)

    Jarullah, B M S; Subramanian, R B; Jummanah, M S J

    2005-01-01

    A number of phylogenetic studies have been carried out on biocontrol agents having similar biological control activity. However, no work has been carried out to determine the phylogenetic relationship amongst various groups of biological control agents with varied biocontrol properties. Our aim was to derive a phylogenetic relationship between diverse biocontrol agents belonging to the deuteromycetes and determine its correlation with their spore morphology and their biocontrol activity. RAPD was used to assess genomic variability in fungi used as biological control agents which included ten isolates of nematophagous fungi such as Arthrobotrys sp., Duddingtonia sp., Paecilomyces sp. and Verticillium sp., along with two isolates of fungal biocontrol agents such as Trichoderma sp. and two isolates of entomopathogenic fungi including Beauveria sp. A plant pathogenic fungus, Verticillium alboatrum was also included to increase the diversity of Deuteromycetes used. A similarity matrix was created using Jaccard's similarity coefficient & clustering was done using unweighted pair group arithmetic mean method (UPGMA). The final dendogram was created using a combination of two programs, Freetree and TreeExplorer. The phylogenetic tree constructed from the RAPD data showed marked genetic variability among different strains of the same species. The spore morphologies of all these fungi were also studied. The phylogenetic pattern could be correlated with the conidial and conidiophore morphology, a criterion commonly used for the classification of fungi in general and Deuteromycetes in particular. Interestingly, the inferred phylogeny showed no significant grouping based on either their biological control properties or the trapping structures amongst the nematophagous fungi as reported earlier by other workers. The phylogenetic pattern was also similar to the tree obtained by comparing the 18S rRNA sequences from the database. The result clearly indicates that the classical

  1. Assessment of genetic diversity in Trigonella foenum-graecum and Trigonella caerulea using ISSR and RAPD markers

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    Ranjekar Prabhakar K

    2004-07-01

    Full Text Available Abstract Background Various species of genus Trigonella are important from medical and culinary aspect. Among these, Trigonella foenum-graecum is commonly grown as a vegetable. This anti-diabetic herb can lower blood glucose and cholesterol levels. Another species, Trigonella caerulea is used as food in the form of young seedlings. This herb is also used in cheese making. However, little is known about the genetic variation present in these species. In this report we describe the use of ISSR and RAPD markers to study genetic diversity in both, Trigonella foenum-graecum and Trigonella caerulea. Results Seventeen accessions of Trigonella foenum-graecum and nine accessions of Trigonella caerulea representing various countries were analyzed using ISSR and RAPD markers. Genetic diversity parameters (average number of alleles per polymorphic locus, percent polymorphism, average heterozygosity and marker index were calculated for ISSR, RAPD and ISSR+RAPD approaches in both the species. Dendrograms were constructed using UPGMA algorithm based on the similarity index values for both Trigonella foenum-graecum and Trigonella caerulea. The UPGMA analysis showed that plants from different geographical regions were distributed in different groups in both the species. In Trigonella foenum-graecum accessions from Pakistan and Afghanistan were grouped together in one cluster but accessions from India and Nepal were grouped together in another cluster. However, in both the species accessions from Turkey did not group together and fell in different clusters. Conclusions Based on genetic similarity indices, higher diversity was observed in Trigonella caerulea as compared to Trigonella foenum-graecum. The genetic similarity matrices generated by ISSR and RAPD markers in both species were highly correlated (r = 0.78 at p = 0.001 for Trigonella foenum-graecum and r = 0.98 at p = 0.001 for Trigonella caerulea indicating congruence between these two systems

  2. Population genetic structure and diversity of high value vulnerable medicinal plant Acorus calamus in India using RAPD and chloroplast microsatellite markers

    Institute of Scientific and Technical Information of China (English)

    H. S. Ginwal; Neha Mittal; Arvind Tomar; V. K. Varshney

    2011-01-01

    Acorus calamus is a highly valued medicinal plant with globaldistribution used in several drugs of health care systems. We evaluatedthe genetic diversity and population structure of 50 populations of A.calamus from different geographical regions in India through RAPD andchloroplast microsatellite markers. From the total screened 82 RAPDprimers and 18 cpSSR primers, 10 RAPD and nine cpSSRs were foundpolymorphic. The selected 10 RAPD primers produced a total of 96reproducible bands, out of which 65 were polymorphic (67.70%).Whereas, the selected nine cpSSR markers produced 26 alleles and all ofthem were polymorphic. The mean genetic diversity (H) among popula-tions using RAPD (H= 0.263) and cpSSR (H=0.530) markers washigher in comparison to the mean genetic diversity within populations.Mean coefficient of gene differentiation (G) between the populationswas also high for both RAPD (G=0.830) and cpSSR markers (G=0.735), whereas the estimated gene flow was very low for RAPD (Nm =0.102) and for cpSSR (Nm = 0.179). AMOVA analysis revealed thatmore genetic variation resided among the populations than within popu-lations. Significant differences (p < 0.001) were observed between thepopulations and individuals within the populations. Cluster analysis ofRAPD and cpSSR data using UPGMA algorithm based on Nei's geneticsimilarity matrix placed the 50 populations into two main clusters. Theimplication of the results of this study in devising strategy for conserva-tion of A. Calamus is discussed.

  3. Taxonomic considerations among and within some Egyptian taxa of Capparis and related genera (Capparaceae as revealed by RAPD fingerprinting

    Directory of Open Access Journals (Sweden)

    El-Bous, M.

    2011-12-01

    Full Text Available Taxonomic considerations among and within some Egyptian taxa of Capparis and related genera (Capparaceae as revealed by RAPD fingerprinting.- This investigation was carried out to assess the taxonomic relationships among eight taxa of the Egyptian members of Capparaceae based on random amplified polymorphic DNA markers, and to compare the results with those obtained from morphological studies. A total of 46 bands were scored for three RAPD primers corresponding to an average of 15.3 bands per primer. The three primers (A03, A07 and A09 revealed eight polymorphic RAPD markers among the studied taxa ranging in size from 200 bp to 1000 bp. Jaccard’s coefficient of similarity varied from 0.28 to 0.84, indicative of high level of genetic variation among the genotypes studied. UPGMA cluster analysis indicated three distinct clusters, one comprised Cleome amblyocarpa and Gynandropsis gynandra, while another included two clusters at 0.74 phenon line; one for Capparis decidua, and the other for Capparis sinaica and all varieties of Capparis spinosa. The four varieties of Capparis spinosa were segregated at 0.84 phenon line. However, one of these varieties was more closely related to Capparis sinaica than to the other three varieties of C. spinosa. The RAPD analysis reported here confirms previous studies based on morphological markers.Consideraciones taxonómicas sobre algunos taxones egipcios de Capparis y géneros relacionados (Capparaceae a partir de RAPDs.- El objetivo de este trabajo es investigar las relaciones taxonómicas entre ocho taxones pertenecientes a las Capparaceae en base a marcadores de tipo RAPD, y comparar los resultados con los obtenidos previamente en estudios morfológicos. Se han contabilizado un total de 46 bandas para tres pares de cebadores, con una media de 15,3 bandas por cebador. Los tres pares de cebadores (A03, A07 y A09 revelan ocho marcadores polimórficos entre los taxones estudiados, de entre 200 y 1000 pares de

  4. Molecular Characterization of Black Bengal and Jamuna Pari Goat Breeds By RAPD Markers

    Directory of Open Access Journals (Sweden)

    M. A. Rahman

    2006-01-01

    Full Text Available This experiment was conducted in the Genetics, Breeding and Reproductive Biotechnology Laboratory under Goat and Sheep Production Research Division, Bangladesh Livestock Research Institute (BLRI, Saver, Dhaka, Bangladesh. DNA was extracted from 14 goat breeds and the extracted DNA was observed by gel electrophoresis. Eight goat specific primers were synthesized by ASM-800 DNA synthesizer and screened in the study and all these primers were capable of priming polymorphic amplification pattern in both the breeds. Random amplification of polymorphic DNA – Polymerase Chain Reaction (RAPD-PCR analysis was carried out using DNA samples of 14 black bengal goat and Jamuna pari goat breeds. Only unambiguous, reproducible and scorable polymorphic fragments were taken into consideration for analysis. Data were analyzed by using a computer programe POPGENE (Version 1.31. Highest level of Nei's[1] gene diversity value (0.4898 was observed in BMS 1494 locus and the mean genetic diversity was obtained 0.3724 among the 14 goat breeds. The highest number of polymorphism obserbed in primer BM1818.The pair-wise genetic distance value ranged from 0.2500 to 1.000. Dendrogram based on Nei's[1] genetic distance using Unweighted Pair Group Method of Arithmetic Means (UPGMA indicated segregation of the 14 goat breeds. Within Jamuna pari goat genetic similarity is low as well as black Bengal goat.

  5. Intra and inter populational genetic variability in Maytenus ilicifolia Mart. ex Reiss. 1861, through RAPD markers

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    AJ. Mossi

    Full Text Available Maytenus ilicifolia is a medicinal plant largely used in the South Brazilian folk medicine. The aim of this study was to quantify the intra and inter populational genetic variability in three populations of M. ilicifolia, focusing on the genetic conservation of this species, which has been threatened by anthropic action. RAPD (Random Amplified Polymorphic DNA markers were used to analyze 30 plants of each of the three populations collected in the Alto Uruguai Gaúcho region. Fourteen selected primers generated a total of 158 bands, 71.5% of which were polymorphic. The comparison of Jaccard’s distances showed that the intra populational variation was higher than the inter populational variability, and cluster analysis allowed the separation of the three populations. Just 7.6% of the bands were specific of at least two populations. Data indicate that the analyzed M. ilicifolia populations represent a single genetic pool, and therefore any of the population thoroughly can represent the overall genetic variability of the species in the sampled region.

  6. Genetic variation in the vulnerable and endemic Monkey Puzzle tree, detected using RAPDs.

    Science.gov (United States)

    Bekessy, Sarah A; Allnutt, T R; Premoli, A C; Lara, A; Ennos, R A; Burgman, M A; Cortes, M; Newton, A C

    2002-04-01

    Araucaria araucana (Monkey Puzzle), a southern South American tree species of exceptional cultural and economic importance, is of conservation concern owing to extensive historical clearance and current human pressures. Random amplified polymorphic DNA (RAPD) markers were used to characterise genetic heterogeneity within and among 13 populations of this species from throughout its natural range. Extensive genetic variability was detected and partitioned by analysis of molecular variance, with the majority of variation existing within populations (87.2%), but significant differentiation was recorded among populations (12.8%). Estimates of Shannon's genetic diversity and percent polymorphism were relatively high for all populations and provide no evidence for a major reduction in genetic diversity from historical events, such as glaciation. All pairwise genetic distance values derived from analysis of molecular variance (Phi(ST)) were significant when individual pairs of populations were compared. Although populations are geographically divided into Chilean Coastal, Chilean Andes and Argentinean regions, this grouping explained only 1.77% of the total variation. Within Andean groups there was evidence of a trend of genetic distance with increasing latitude, and clustering of populations across the Andes, suggesting postglacial migration routes from multiple refugia. Implications of these results for the conservation and use of the genetic resource of this species are discussed.

  7. RAPD analysis of the genetic diversity of mango (Mangifera indica) germplasm in Brazil.

    Science.gov (United States)

    Souza, I G B; Valente, S E S; Britto, F B; de Souza, V A B; Lima, P S C

    2011-12-14

    We evaluated genetic variability of mango (Mangifera indica) accessions maintained in the Active Germplasm Bank of Embrapa Meio-Norte in Teresina, Piauí, Brazil, using RAPDs. Among these accessions, 35 originated from plantings in Brazil, six from the USA and one from India. Genomic DNA, extracted from leaf material using a commercial purification kit, was subjected to PCR with the primers A01, A09, G03, G10, N05, and M16. Fifty-five polymorphic loci were identified, with mean of 9.16 ± 3.31 bands per primer and 100% polymorphism. Application of unweighted pair group method using arithmetic average cluster analysis demonstrated five genotypic groups among the accessions examined. The genotypes Rosa 41, Rosa 48 and Rosa 49 were highly similar (94% similarity), whereas genotypes Sensation and Rosa 18 were the most divergent (only 7% similarity). The mango accessions were found to have considerable genetic variability, demonstrating the importance of analyzing each genotype in a collection in order to efficiently maintain the germplasm collection.

  8. Characterization of the Second Generation Cryopreserved Dendrobium Bobby Messina Using Histological and RAPD Analyses

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    Jessica Jeyanthi James Antony

    2015-12-01

    Full Text Available This study was conducted to detect the morphological, histological and molecular diff erences in the second generation of the PVS2 cryopreserved Dendrobium Bobby Messina [DBM] (18 months old culture plantlets. Morphological analyses indicated that similarities and diff erences in cryopreserved DBM plantlets comparing to control stock culture based on selected morphological criteria. Morphological criteria, such as root length, number of shoot per explant and shoot length displayed diff erences, while the other three criteria, leaf diameter, leaf length and PLBs size were similar in cryopreserved compared to the control stock culture plant. Higher amount of homogenous cell population and denser cytoplasm were observed in cryopreserved PLBs compared to control stock culture PLBs based on histological analysis. This suggests the existance of somatic embryogenesis development mechanism taking place during the recovery and regeneration of the cryopreserved PLBs. However, RAPD analyses based on 10 primers indicated that cryopreserved DBM regenerated from vitrifi cation method generated a total of 20 to 39.9% polymorphic bands as compared to stock culture indicating potential somaclonal variation. Hence, an increase percentage of polymorphics bands in cryopreserved plantlets 18 months post cryopreservation as compared to previous report of 10% polymorphic bands in cryopreserved DBM 3 months post cryopreservation.

  9. Genetic Diversity of Dalmatian Sage (Salvia officinalis L. as Assessed by RAPD Markers

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    Zlatko Liber

    2014-09-01

    Full Text Available Dalmatian or common sage (Salvia officinalis L. is an outcrossing plant species native to East Adriatic coast. Random Amplified Polymorphic DNA markers (RAPD were used to analyze genetic diversity and structure of ten natural populations from the East-Adriatic coastal region. The highest genetic diversity was found in populations from the central and south Dalmatia, while the highest frequency down-weighted marker values were found in the northernmost populations and the southern most inland population. Although analysis of molecular variance (AMOVA revealed that most of the genetic diversity was attributable to differences among individuals within populations, highly significant φST values suggested the existence of genetic differentiation among populations. By assuming Hardy-Weinberg equilibrium within populations, the calculated FST value among population was moderate. Bayesian model-based clustering method revealed that at K = 2 all individuals belonging to two northern populations were assigned to a separate cluster from the individuals belonging to the rest of the population. At K = 3, the newly formed cluster grouped the majority of individuals belonging to populations from central Dalmatia. The high correlation between matrices of genetic and geographical distances showed that isolation by distance may play a considerable role in overall structuring of the genetic diversity.

  10. Genetic Diversity of Dalmatian Sage (Salvia offi cinalis L. as Assessed by RAPD Markers

    Directory of Open Access Journals (Sweden)

    Zlatko Liber

    2014-10-01

    Full Text Available Dalmatian or common sage (Salvia officinalis L. is an outcrossing plant species native to East Adriatic coast. Random Amplified Polymorphic DNA markers (RAPD were used to analyze genetic diversity and structure of ten natural populations from the East-Adriatic coastal region. The highest genetic diversity was found in populations from the central and south Dalmatia, while the highest frequency down-weighted marker values were found in the northernmost populations and the southern most inland population. Although analysis of molecular variance (AMOVA revealed that most of the genetic diversity was attributable to differences among individuals within populations, highly significant φST values suggested the existence of genetic differentiation among populations. By assuming Hardy-Weinberg equilibrium within populations, the calculated FST value among population was moderate. Bayesian model-based clustering method revealed that at K = 2 all individuals belonging to two northern populations were assigned to a separate cluster from the individuals belonging to the rest of the population. At K = 3, the newly formed cluster grouped the majority of individuals belonging to populations from central Dalmatia. The high correlation between matrices of genetic and geographical distances showed that isolation by distance may play a considerable role in overall structuring of the genetic diversity.

  11. Genetic diversity analysis by RAPD in Cathaya argyrophylla Chun et Kuang

    Institute of Scientific and Technical Information of China (English)

    汪小全; 邹喻苹; 张大明; 洪德元; 刘正宇

    1997-01-01

    Genetic diversity level of Cathaya argyrophylla was confirmed by random amplified polymorphic DNA (RAPD) markers. Seventy five samples (individuals), collected from Hunan and Sichuan provinces of China were used in the study. 21 10-mer oligonucleotide primers detected 106 sites, and 34 (32% ) of them were polymor-phic. The level of genetic variation in C. argyrophylla was lower than those of other conifers, and was considered to be associated with the complexity of habitats. The percentages of polymorphic sites (PPS) in the Hunan and Sichuan pop-ulations were 18% and 25% respectively. 7.99% of genetic variation existed between the two populations; this value was higher than the mean value (6.8%) among populations in conifers displayed by allozyme. Some subpopulations of C. argyrophylla were greatly differentiated because of site mutation and genetic drift. The highest value of genetic dif-ference between subpopulations amounted to 16. 23% . In addition, a concept of diversity coefficient(DC), a value us

  12. Detection and Identification of Probiotic Lactobacillus plantarum Strains by Multiplex PCR Using RAPD-Derived Primers

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    Alex Galanis

    2015-10-01

    Full Text Available Lactobacillus plantarum 2035 and Lactobacillus plantarum ACA-DC 2640 are two lactic acid bacteria (LAB strains that have been isolated from Feta cheese. Both display significant potential for the production of novel probiotic food products. The aim of the present study was the development of an accurate and efficient method for the molecular detection and identification of the above strains in a single reaction. A multiplex PCR assay was designed for each strain, based on specific primers derived from Random Amplified Polymorphic DNA (RAPD Sequenced Characterized Amplified Region (SCAR analysis. The specificity of the assay was tested with a total of 23 different LAB strains, for L. plantarum 2035 and L. plantarum ACA-DC 2640. The multiplex PCR assay was also successfully applied for the detection of the above cultures in yogurt samples prepared in our lab. The proposed methodology may be applied for monitoring the presence of these strains in food products, thus evaluating their probiotic character. Moreover, our strategy may be adapted for other novel LAB strains with probiotic potential, thus providing a powerful tool for molecular discrimination that could be invaluable to the food industry.

  13. Differential survival of mosquitofish exposed to radionuclides is dependent on RAPD genotype

    Energy Technology Data Exchange (ETDEWEB)

    Theodorakis, C.W. [Oak Ridge Associated Universities, TN (United States); Shugart, L.R. [Oak Ridge National Lab., TN (United States)

    1995-12-31

    In previous studies, it was found that certain RAPD (Randomly Amplified Polymorphic DNA) markers were present at higher frequencies in radionuclide-contaminated mosquitofish (Gambusia affinis) populations than in reference populations. These markers will be referred to as contaminant specific markers. In the present study, fish with and without these markers were collected from non-contaminated populations and exposed in situ to radionuclides by caging them in one of the contaminated sites. Forty fish were exposed for 1--6 weeks, after which the survivors were collected and DNA was extracted for genotypic analysis. In one experiment, the frequencies of contaminant specific markers in the survivors were compared to the frequencies of these markers in the native contaminated and uncontaminated (the source of the caged fish) populations. It was found that the genotypic distributions were more similar to the native contaminated population. In another experiment, samples of caudal fin tissue were collected for DNA extraction before and after placing fish in the cages, in order to compare survival rates of different genotypes. It was found that fish with the contaminant indicative bands had higher percent survival than the other fish. Experiments are underway or are being planned in order to determine the molecular identity of these bands and the ecological significance of altered band frequencies in hopes of developing population-level biomarkers of contaminant exposure and ecological affects.

  14. Separation and relationship of ten marine scuticociliates (Protozoa, Ciliophora) using RAPD fingerprinting method

    Institute of Scientific and Technical Information of China (English)

    SHANG Huimin; SONG Weibo

    2005-01-01

    On the basis of the random amplified polymorphic DNA-fingerprinting (RAPD) method, 10 morphospecies of scuticociliates from 7genera, including 15 clones of 13 strains, Uronema rnarinum, Uronema cf marinum, Parauronerna virginianum, P. longurn, Metanophrys similis, M. sinensis, Paralembus digitiformis, Mesanophrys carcini, Paranophrys magna and Cohnilembus verminus were analysed using 8 oligonucleotide primers. The genetic similarity among the clones of the same strain measured by a band-sharing index is 0.97~0.98,while 0.40~0.52 among strains. This value measured is 0.39~0.46 among congeners of the same genus, whereas 0.16~0.47 between different genera. A distance tree was constructed based on 8-primer analysis, in which the scuticociliates investigated were separated into two clusters: one consists of 2 genera, Uronema and Parauronema, and the other was composed of five, Metanophrys, Mesanophrys,Paranophrys, Paralembus and Cohnilembus. It is demonstrated also that the morphospecies Parauronema virginianum may be a species-complex, i. e., it contains different genospecies.

  15. Assessment of genetic diversity in Dalbergia sissoo clones through RAPD profiling

    Institute of Scientific and Technical Information of China (English)

    Meena Bakshi; Arvind Sharma

    2011-01-01

    We studied the genetic polymorphism among 29 clones of shisham (Dalbergia sissoo Roxb) belonging to different geographic regions using random amplified polymorphic DNA (RAPD) markers. Out of 30 primers used, only 20 primers generated polymorphism in amplified product. In total 232 bands were amplified with 20 primers, of which 192 (82%) were polymorphic with an average of 9.6 bands/primer. The resolving power (Rp) ranged from 2.14 (Primer 5) to 11.93 (Primer 4). Primer 4 and Primer 3 possessed high Rp value. Polymorphism in- formation content (PIC) ranged from 0.15 (Primer 5) to 0.37 (Primer 4). Primer 4 amplified total 18 bands in 29 genotypes with PIC value of 0.37 hence; this set of primer was most informative. The similarity co- efficient analysis revealed two clusters. The first cluster comprised of only 10 clones and the second major cluster comprised of 19 clones. The genetic similarity among 29 clones ranged from 25.86% (clone 10 and 235) to 100% (clone 19 and 59), suggesting a wide genetic base in shisham clones.

  16. Micropropagation of loblolly pine by somatic organogenesis and RAPD analysis of regenerated plantlets

    Institute of Scientific and Technical Information of China (English)

    唐巍

    2000-01-01

    Organogenesis was induced in callus derived from mature zygotic embryos of six families (J-56, S-1003, E-22, E-311, E-440, and Mc) of loblolly pine (Pinus taeda L.) within 24 weeks of culture. Elongation of adventitious buds was achieved on TE medium supplemented with 0.5 mg·L-1 indole-3-butyric acid (IBA) and 1 mg/l 6-benzyladenine (BA). The most suitable medium for root formation proved to be TE medium supplemented with 0.5 mg·L-1 IBA, 2 mg·L-1 BA , and 0.5 mg/l gibberellic acid (GA3). 169 regenerated plantlets were transferred to a perlite∶peatmoss∶vermiculite (1:1:1) soil mixture, and 98 plantlets survived in the field. Total DNA was extracted from the needles of the regenerated plantlets of the six families of loblolly pine. Analysis of random amplified polymorphic DNA (RAPD) using 20 arbitrary oligonucleotide 10-mers, show that amplification products were monomorphic for all the plantlets of family J-56, S-1003, E-22, E-311, E-440, and Mc of loblolly pine. These results suggested that organogenesis can be used for clonal micropropagation of some families of loblolly pine.

  17. Introduction of exogenous wild soybean DNA into cultivated soybean and RAPD molecular verification

    Institute of Scientific and Technical Information of China (English)

    谢纬武; 王斌; 雷勃钧; 李希臣; 卢翠华; 钱华; 周思君

    1995-01-01

    The exogenous total DNA of the wild high-protein soybean was transferred to cultivatedsoybean through the pollen tube channel and the genomic variation of the transformed progeny was detected bythe method of RAPD(Random Amplified Polymorphic DNA).Distinguished variations were found in one of the 7 transformed plants of the first generation(D1),ofwhich the traits of fruition,outward appearance,leaf shape and flower colour were almost identical withthose of the recipient parent;of which grain weight,seed coat colour and stem strength were situated betweenthe two parents;and there were greatly more pods per plant and 12.5% higher content of protein in seedsthan that of the recipient parent.All the properties have been invariably inherited for 3 generations.Through RAPD analysis of the genomes of the donor,the recipient and the transformed progeny(D3)as agroup,DNA polyrnorphisms were found in amplified products by 24 of 150 primers.The results prove thatthe exogenous DNA caused the distinct variance of the genome.The authors infer that the homogeneousrecombination of large exogenous DNA is the main cause for the variance.

  18. Genetic Analysis of Pinus sylvestris L. and Pinus sylvestris forma turfosa L. Using RAPD Markers

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    Beáta ÁBRAHÁM

    2010-03-01

    Full Text Available The purpose of the present study was to determine the level of genetic diversity within and among Ciuc basin, Romania (populations from Mohos and Luci raised bogs in Harghita Mountain and Sumuleu in Ciuc Mountain Pinus sylvestris populations using molecular markers. Two of populations (Mohos and Luci seems to be the descendants that survived the continental glaciation. Genetic diversity was analyzed by RAPD (Random Amplified Polymorphic DNA. Nine primers were selected for analysis, which generated reproducible bands. On base of presence or absence of homologues bands Nei’s gene diversity, the percentage of polymorphic loci and Nei’s unbiased genetic distance were calculated. The level of genetic variation among populations was found to be low. For both populations the variation values among populations were higher than within populations. The fossil records and geological historical data explain the extremely low genetic diversity of this species. Pinus sylvestris experienced strong bottlenecks during its evolutionary history, which caused the loss of genetic variation. Genetic drift and breeding in post-bottlenecked small populations may be the major forces that contribute to low genetic diversity and genetic differentiation of populations. Human activities may have accelerated the loss of genetic diversity in Pinus sylvestris.

  19. Genetic Diversity of Parkia biglobosa from Different Agroecological Zones of Nigeria Using RAPD Markers

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    Oluwafemi Amusa

    2014-01-01

    Full Text Available Parkia biglobosa (Jacq. is an important leguminous tree crop in the African Savannahs useful to the natives where it is found, for domestic use. Previous diversity studies on this tree crop had been majorly on morphological and biochemical analysis. In order to capture the maximum diversity not obtained by previous research, the study aimed at evaluating the genetic diversity of accessions of this crop in the different agroecological zones in Nigeria using RAPD markers. A total of 81 scorable bands with an average of 8.1 bands per primer were amplified among the accessions studied. Intrazonal genetic diversity analysis showed a percentage polymorphism with a range of 11.11% to 65.43% among the agroecological zones studied. Although, gene diversity was highest within Humid forest agroecological zone, a low genetic distance and high genetic similarity between the agroecological zones were observed. Cluster analysis indicated six main groups of which four groups had single accessions while the two groups clustered the remaining accessions, indicating a narrowed genetic base from the 23 accessions studied.

  20. Detection and Identification of Probiotic Lactobacillus plantarum Strains by Multiplex PCR Using RAPD-Derived Primers.

    Science.gov (United States)

    Galanis, Alex; Kourkoutas, Yiannis; Tassou, Chrysoula C; Chorianopoulos, Nikos

    2015-10-22

    Lactobacillus plantarum 2035 and Lactobacillus plantarum ACA-DC 2640 are two lactic acid bacteria (LAB) strains that have been isolated from Feta cheese. Both display significant potential for the production of novel probiotic food products. The aim of the present study was the development of an accurate and efficient method for the molecular detection and identification of the above strains in a single reaction. A multiplex PCR assay was designed for each strain, based on specific primers derived from Random Amplified Polymorphic DNA (RAPD) Sequenced Characterized Amplified Region (SCAR) analysis. The specificity of the assay was tested with a total of 23 different LAB strains, for L. plantarum 2035 and L. plantarum ACA-DC 2640. The multiplex PCR assay was also successfully applied for the detection of the above cultures in yogurt samples prepared in our lab. The proposed methodology may be applied for monitoring the presence of these strains in food products, thus evaluating their probiotic character. Moreover, our strategy may be adapted for other novel LAB strains with probiotic potential, thus providing a powerful tool for molecular discrimination that could be invaluable to the food industry.

  1. The FML (Fucose Mannose Ligand of Leishmania donovani: a new tool in diagnosis, prognosis, transfusional control and vaccination against human kala-azar

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    Claris B. Palatnik de Sousa

    1996-04-01

    Full Text Available The Fucose-Mannose Ligand (FML of Leishmania donovani is a complex glycoproteic fraction. Its potential use as a tool for diagnosis of human visceral leishmaniasis was tested with human sera from Natal, Rio Grande do Norte, Brazil. The FML-ELISA test, showed 100% sensitivity and 96% specificity, identifying patients with overt kala-azar (p O FML (Ligame de Fucose-Manose de Leishmania donovani é uma fração glicoproteica complexa. O seu potencial no diagnóstico da leishmaniose visceral humana foi testado com soros provenientes de Natal, Rio Grande do Norte, Brasil. O teste de FML-ELISA mostrou 100% de sensibilidade e 96% de especificidade, identificando pacientes com calazar declarado (p<0.001, comparados com soros normais e indivíduos com infecção subclínica. Mais de 20% dos sororreativos assimptomáticos desenvolveram a doença no prazo de 10 meses. Na análise de doadores de sangue, 5% de sororeativos, atingindo até 17% num único dia foram detectados. A glicoproteínaGP36 do FHL é reconhecida especificamente por soros de pacientes com calazar. O potencial imunoprotetor do FML no calazar experimental foi testado no modelo swiss albino em combinação com saponina pelas vias subcutâneas e/ou intraperitoneal seguido de desafio com 2x 10(7 amastigolas de Leishmania donovani. Um aumento de 80.0% na resposta de anticorpos específicos (p<0.001 e a redução de 85.5 % da carga parasitária no fígado (p<0.001 foi detectado nos animais vacinados com FML e saponina, independentemente da via de administração.

  2. A curative immune profile one week after treatment of Indian kala-azar patients predicts success with a short-course liposomal amphotericin B therapy.

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    Smriti Mondal

    Full Text Available BACKGROUND: The present pilot study investigating the minimum dose for short-course single and double-dose treatment of kala-azar with an apparently new liposomal formulation of amphotericin B, Fungisome, led to identification of immunological components for early detection of success and/or failure to cure. METHODS: Patients were treated with 5, 7.5 (single-dose and 10 mg/kg body weight (5 mg/kg double-dose of Fungisome. Immunological investigations involving plasma cytokines and antigen-specific lymphoproliferation and cytokine responses from PBMCs were carried out before, 1 week after Fungisome treatment, at the time of relapse, and again after conventional amphotericin B treatment. RESULTS: At 1-month follow-up all the patients showed 100% initial cure. However, total doses of 5, 7.5 and 10 mg/kg Fungisome showed 60%, 50% and 90% cure, respectively, at 6-months posttreatment. Patients successfully cured demonstrated downregulation of IL-12 and IL-10 in plasma, and two-fold or more elevation of IFN-gamma, IL-12 and TNF, and significant down-regulation of IL-10 and TGF-beta in culture supernatants 1-week posttreatment irrespective of drug-dose. A differential immune profile, involving insignificant decline in IL-10 and IL-12 in plasma and negligible elevation of IFN-gamma, IL-12 and TNF, and persistence of IL-10, despite decline in TGF-beta in culture supernatants, in apparently cured individuals, corresponded with relapse within 6-months of treatment. CONCLUSION: Immunological investigations revealed significant curative and non-curative immunomodulation 1-week posttreatment, correlating with successful cure and relapse, respectively. Although immune-correlation was dose-independent, almost consistent curative response in patients treated with the highest dose 10 mg/kg reflected a definitive impact of the higher-dose on the immune response. TRIAL REGISTRATION NAME AND NUMBER: Clinical Trials Registry--India (CTRI CTRI/2009/091/000764.

  3. Análise da recuperação do genitor recorrente em maracujazeiro-azedo por meio de marcadores RAPD Recovery analysis of recurrent genitor in sour passion fruit through RAPD markers

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    Kenia Gracielle da Fonseca

    2009-03-01

    Full Text Available O Brasil é o maior produtor mundial de maracujá, entretanto tem-se observado redução na produtividade do maracujazeiro nos últimos anos, devido, principalmente, a fatores fitossanitários. Na Embrapa Cerrados, a transferência de genes de resistência de espécies silvestres para as comerciais de maracujazeiro tem sido feita por meio de hibridações interespecíficas seguidas de um programa de retrocruzamentos auxiliados por marcadores moleculares. Este trabalho teve por objetivo verificar a recuperação do genoma recorrente nas plantas RC4 e RC5 [(Passiflora edulis x Passiflora setacea x Passiflora edulis ] com base em marcadores RAPD. O estudo foi desenvolvido no Laboratório de Genética e Biologia Molecular da Embrapa Cerrados. Amostras de DNA de cada material genético (17 plantas RC4, 16 plantas RC5, Passiflora edulis e Passiflora setacea foram amplificadas para obtenção de marcadores RAPD. Foram utilizados 12 primers decâmeros para as plantas RC4 e 14 primers decâmeros para as plantas RC5. Os marcadores RAPD gerados foram convertidos em matriz de dados binários. Verificou-se alta porcentagem de marcadores polimórficos em consequência do cruzamento-base interespecífico. A menor similaridade genética foi observada entre as espécies P. edulis e P. setacea, evidenciando a grande distância genética dessas espécies.Brazil is the largest world producer of passion fruit, however, it has been observed a reduction in the productivity in recent years due, mainly, to phytosanitary factors. At Embrapa Cerrados, the transfer of resistance genes from wild to commercial species of passion fruit has been made through interspecific hybridations, followed by a backcrossing molecular marker-assisted program. The objective this work was to verify the recovery of recurrent genome at the plants RC4 and RC5 [(Passiflora edulis x Passiflora setacea x Passiflora edulis] based on RAPD markers. The study was developed at Embrapa Cerrados

  4. Genetic variability of Streptococcus mutans isolated from low-income families, as shown by RAPD markers Variabilidade genética de Streptococcus mutans isolados de famílias de baixa renda, demonstrados por marcadores RAPD

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    Mônica Moreira

    2007-12-01

    Full Text Available The detection of Streptococcus mutans isolates with high genetic variability in different individuals indicates the occurrence of transmissibility. In this context, nine low-income families (40 individuals in total with similar social conditions were assessed to identify the bioserotypes of S. mutans using both biochemical and RAPD markers and to establish the degree of similarity among the intra-familial isolates. The polymorphism analysis used the coefficient of Jaccard in both a Principal Coordinates Analysis (PCO and in a UPGMA clustering method. A total of 157 isolates were obtained from salivary samples, using the morphology of colonies recovered using MSB agar as indicators. From those, 64 were characterized biochemically as S. mutans and 10 as S. sombrinus. Genetic variability among isolates based on RAPD markers was not consistent with their intra-familial distribution. In particular, some individuals might have experienced multiple infections given the high genetic variability among their isolates. The occurrence of 4 isolates with 100% genetic similarity is indicative of intra-familial transmission.A detecção de isolados de S. mutans com alta similaridade genética em indivíduos diferentes, sugere a existência de transmissibilidade. Neste contexto, nove famílias (40 indivíduos de baixo poder aquisitivo com condições sociais homogêneas foram avaliadas, visando identificar os biosorotipos de S. mutans por meio de bioquimismo e marcadores RAPD e estabelecer o grau de similaridade entre os isolados intra-familiar. Para a análise de polimorfismo utilizou-se coeficiente de Jaccard, análise de coordenadas principais (PCO e método "UPGMA". Foram obtidos 157 isolados, a partir de amostras salivares, usando como indicador a morfologia das colônias recuperadas em ágar MSB. Destes, 64 foram caracterizados bioquimicamente como S. mutans e 10 como S. sobrinus. A partir dos marcadores RAPD verificou-se variabilidade genética entre

  5. Diversidade genética de cultivares de alho (Allium sativum L. por meio de marcador molecular rapd Genetic diversity of the cultivars of garlic (Allium sativum L. for molecular marker rapd

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    José Hortêncio Mota

    2004-08-01

    Full Text Available Realizou-se este estudo com o objetivo de determinar a diversidade genética entre doze cultivares de alho, por meio de marcadores moleculares RAPD. Foram utilizados seis cultivares nobres e seis cultivares seminobres. A análise de agrupamento das similaridades genéticas foi realizada pelo método UPGMA, gerando um dendrograma utilizando o índice de Jaccard. Houve a formação de dois grupos, sendo o primeiro grupo formado pelas cultivares nobres (Roxo Pérola de Caçador, Chonan, Contestado 12, Caçador 30, Caçador 40 e Quitéria 595, ou seja, cultivares que precisam de vernalização para a formação do bulbo, e um segundo formado pelas cultivares seminobres (Gigante Curitibanos, Gigante Roxo, Gigante Roxão, Gravatá, Amarante e Cateto Roxo ou que não precisam de vernalização para a formação do bulbo. As cultivares nobres e seminobres apresentaram 57,1% e 54,2% de similaridade, respectivamente. Pelos resultados, pode-se concluir que o marcador molecular RAPD foi eficiente em separar dois grupos de cultivares de Allium sativum.The objective of this study was to determine the genetic diversity among twelve garlic cultivars, with RAPD molecular marker. Six cultivars noble and six cultivars half-noble were tested. The analysis of grouping by genetic similarities was carried out by the method of UPGMA with generated a dendrogram using the Jaccard index. Have the formation of two groups, being the first group formed by the cultivars noble (Roxo Pérola de Caçador, Chonan, Contestado 12, Caçador 30, Caçador 40 and Quitéria 595 those that need vernalization for the formation of the bulb and a second group formed by the cultivars half-noble (Gigante Curitibanos, Gigante Roxo, Gigante Roxão, Gravatá, Amarante and Cateto Roxo which do not need vernalization for formation of the bulb. The cultivars noble and half-noble presented 57,1% and 54,2% of similarity, respectively. The results allowed to conclued that RAPD molecular marker were

  6. Genetic variations among passion fruit species using rapd markers Variação genética entre espécies de maracujá utilizando marcadores rapd

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    Ana Paula de Andrade Aukar

    2002-12-01

    Full Text Available It has been evaluated the genetic variability through the use of RAPD molecular markers on the following passionflower species: Passiflora amethystina, P. caerulea, P. cincinnata, P. coccinea, P. serrato digitata, P. foetida, P. maliformis, P. alata, P. giberti, P. laurifolia, P. macrocarpa, P. nitida, P. setacea, P. suberosa, P. ligularis, P. capsularis, P. edulis Sims and its botanical variety P. edulis Sims f. flavicarpa Deg. In this research work, the analyses of the random amplified polymorphic DNA products (RAPD were employed to estimate the genetic diversity and the taxonomic linkage within the species above. The total of 21 primers were used in this study which generated 270 different polymorphic products. It was possible to detect that the Passiflora species had shown a similarity of 17,3%, and between Passiflora edulis Sims and Passiflora edulis Sims f. flavicarpa a similarity of 34,35% has been found. The rate of similarity within edulis specie is low, making it clear that a large variability between the yellow and the purple forms exists.Foram avaliadas as variações genéticas através de marcadores moleculares RAPD, as seguintes espécies de maracujá: Passiflora amethystina, P. caerulea, P. cincinnata, P. coccinea, P. serrato digitata, P. foetida, P. maliformis, P. alata, P. giberti, P. laurifolia, P. macrocarpa, P. nitida, P. setacea, P. suberosa, P. ligularis, P. capsularis, P. edulis Sims e sua variedade botânica P. edulis Sims f. flavicarpa Deg. Neste estudo, a análise dos produtos da amplificação ao acaso do DNA polimórfico (RAPD foi usada para estimar a diversidade genética e as relações taxonômicas entre as espécies. Foram utilizados 21 "primers", que produziram um total de 270 bandas polimórficas. Verificou-se que as espécies de Passiflora apresentaram uma média de similaridade de 17,3%, e entre Passiflora edulis Sims e Passiflora edulis Sims f. flavicarpa, de 34,35%. Pode-se perceber que o valor de

  7. Genetic diversity in Penaeus chinensis shrimp as revealed by RAPD technique

    Institute of Scientific and Technical Information of China (English)

    庄志猛; 石拓; 孔杰; 刘萍; 刘振辉; 孟宪红; 邓景耀

    2001-01-01

    The random amplified polymorphic DNA analysis was used to estimate genetic diversity in one successively cultivated stock and three wild stocks of Penaeus chinensis shrimp, two of which were collected from the spawning and wintering grounds in the west coast of Korean Peninsula, and one from the feeding ground in the China coast of the Yellow Sea. A random primer kit was employed to scan the genomic DNA in 20 individuals of each index stock. A total of 110 reproducible RAPD markers were obtained, 68.2 % of which showed a sound eonformability within all the individuals detected, implying that the genetic variability in P. chinensis is relatively low. The proportions of polymorphic loci among these four stocks ranged from 20% to 33.3%, while the degrees of genetic polymorphisms varied from 0.0093 to 0.0307. The genetic variability of inter-stocks was higher than that of intra-stock. The genetic diversity in different stocks differed from each other; that is, a less genetic differentiation in the spawning and wintering stocks from the west coast of Korean Peninsula was revealed and their genetic diversities were higher than that of the spawning stock in the Bohai Sea and the China coast of the Yellow Sea. As detected, the genetic diversity in the successively cultivated stock was the lowest among these four stocks. Through genetic distance analysis between a random pair of individuals, a dendrogram of the above-mentioned four stocks was constructed by unweighted pair group method with arithmetic mean. The results based on cluster analysis well fitted with the geographical distribution of P. chinensis in the Bohai and Yellow Seas.

  8. RAPD linkage mapping in a longleaf pine x slash pine F1 family.

    Science.gov (United States)

    Kubisiak, T L; Nelson, C D; Nance, W L; Stine, M

    1995-06-01

    Random amplified polymorphic DNAs (RAPDs) were used to construct linkage maps of the parent of a longleaf pine (Pinus palustris Mill.) slash pine (Pinus elliottii Englm.) F1 family. A total of 247 segregating loci [233 (1∶1), 14 (3∶1)] and 87 polymorphic (between parents), but non-segregating, loci were identified. The 233 loci segregating 1∶1 (testcross configuration) were used to construct parent-specific linkage maps, 132 for the longleaf-pine parent and 101 for the slash-pine parent. The resulting linkage maps consisted of 122 marker loci in 18 groups (three or more loci) and three pairs (1367.5 cM) for longleaf pine, and 91 marker loci in 13 groups and six pairs for slash pine (952.9 cM). Genome size estimates based on two-point linkage data ranged from 2348 to 2392 cM for longleaf pine, and from 2292 to 2372 cM for slash pine. Linkage of 3∶1 loci to testcross loci in each of the parental maps was used to infer further linkages within maps, as well as potentially homologous counterparts between maps. Three of the longleaf-pine linkage groups appear to be potentially homologous counterparts to four different slash-pine linkage groups. The number of heterozygous loci (previously testcross in parents) per F1 individual, ranged from 96 to 130. With the 87 polymorphic, but non-segregating, loci that should also be heterozygous in the F1 progeny, a maximum of 183-217 heterozygous loci could be available for mapping early height growth (EHG) loci and for applying genomic selection in backcross populations.

  9. Development of AFLP and RAPD markers linked to a locus associated with twisted growth in corkscrew willow (Salix matsudana 'Tortuosa').

    Science.gov (United States)

    Lin, Juan; Gunter, Lee E; Harding, Scott A; Kopp, Richard F; McCord, Rachel P; Tsai, Chung-Jui; Tuskan, Gerald A; Smart, Lawrence B

    2007-11-01

    Salix matsudana Koidz. cultivar 'Tortuosa' (corkscrew willow) is characterized by extensive stem bending and curling of leaves. To investigate the genetic basis of this trait, controlled crosses were made between a corkscrew female (S. matsudana 'Tortuosa') and a straight-stemmed, wild-type male (Salix alba L. Clone 99010). Seventy-seven seedlings from this family (ID 99270) were grown in the field for phenotypic observation. Among the progeny, 39 had straight stems and leaves and 38 had bent stems and curled leaves, suggesting that a dominant allele at a single locus controls this phenotype. As a first step in characterizing the locus, we searched for amplified fragment length polymorphism (AFLP) and randomly amplified polymorphic DNA (RAPD) markers linked to the tortuosa allele using bulked segregant analysis. Samples of DNA from 10 corkscrew individuals were combined to produce a corkscrew pool, and DNA from 10 straight progeny was combined to make a wild-type pool. Sixty-four AFLP primer combinations and 640 RAPD primers were screened to identify marker bands amplified from the corkscrew parent and progeny pool, but not from the wild-type parent or progeny pool. An AFLP marker and a RAPD marker linked to and flanking the tortuosa locus were placed on a preliminary linkage map constructed based on segregation among the 77 progeny. Sectioning and analysis of shoot tips revealed that the corkscrew phenotype is associated with vascular cell collapse, smaller cell size in regions near the cambium and less developed phloem fibers than in wild-type progeny. Identification of a gene associated with this trait could lead to greater understanding of the control of normal stem development in woody plants.

  10. [The use of RAPD and ITE molecular markers to study genetical structure of the Crimean population of Triticum boeoticum Boiss].

    Science.gov (United States)

    Mallabaeva, D Sh; Ignatov, A N; Sheĭko, I A; Isikov, V P; Geliuta, V P; Boĭko, N G; Seriapin, A A; Dorokhov, D B

    2007-01-01

    Wild wheat Triticum boeoticum Boiss. is the rare species are included in the Red Book of Ukraine. This species are reducing the magnitude of population and the area of distribution under anthropogenic activity. We studied genetic structure of two populations of T. boeoticum, located on Sapun Mountain and in Baidar Valley in Crimea. According RAPD and ITE molecular analysis we have estimated that the population of T. boeoticum on Sapun Mountain is genetically more impoverished than a population from the Baidar Valley. For preservation of maximal natural genetic polymorphism of the rare species it is recommended to direct efforts to preservations of a population of T. boeoticum from the Baidar Valley.

  11. Establishment and Optimization of RAPD Reaction System of Cordyceps sinensis%冬虫夏草RAPD反应体系的建立及优化

    Institute of Scientific and Technical Information of China (English)

    张德利; 钱敏; 曾纬; 陈仕江

    2011-01-01

    To establish RAPD reaction system of Cordyceps sinensis. Through changing the density of Mg2+,dNTP, primer, template DNA in RAPD reaction system, with the result of RAPD amplify, established the RAPD reaction system of Cordyceps sinensis. Then through changing the main thermal cycle parameter,optimize RAPD reaction system of Cordyceps sinensis. The results showed that an RAPD reaction system suitable for Cordyceps sinensis was established, that was: 20 μL amplification containing 1 ×PCR buffer, 1.5 μmol/μL Mg2+, 320 μmol/L dNTP, 24 ng premer、 20 ng template DNA、 1 U Taq DNA polymerase. The optimization result of amplifying the procedure was that: pre-Modified 5 min at 95℃, and then circulated 35 times (denatured 45 s at 94℃, renatured 1 min at 36℃, elongated 2 min at 72℃), and then elongated 2 min at 72℃ after the cycle. The RAPD reaction system can be used to RAPD analysis in Cordyceps sinensis.%为建立并优化冬虫夏草RAPD反应体系,通过改变RAPD反应体系中Mg2+,dNTP、引物、模板等主要成分的浓度,结合RAPD扩增效果,建立冬虫夏草RAPD反应体系,然后通过改变主要热循环参数.优化冬虫夏草RAPD反应体系.结果表明:适合冬虫夏草的RAPD反应体系为25μL体系中内含1×PCR缓冲液、1.5 μmol/μLMg2+、320 μmaol/L dNTP、24 ng引物、20 ng模板、1 U Taq酶;扩增程序的优化结果为:95℃预变性5 min,然后35个循环(94℃变性45 s,36℃复性1 min,72℃延伸2 min),循环结束后72℃延伸7 min.综上,RAPD技术可用于冬虫夏草的鉴定、评价分析.

  12. RAPD marker and its applications in forage study%RAPD标记技术及其在牧草研究中的应用

    Institute of Scientific and Technical Information of China (English)

    桂枝; 袁庆华; 蔺海明

    2001-01-01

    介绍了RAPD标记的发展、原理、方法、特点和不足,并且对它在遗传图谱的构建、基因定位、品种和品系的分类及鉴定、作物育种等研究领域中的应用作一简述。%This review introduced the evolution, theory, method, characteristics and disadvantages of RAPD analysis. The advance and applications of RAPD marker on genetic link-age mapping, gene localization, population classification and identification and forage breed-ing were described.

  13. El azar y sus problemas

    OpenAIRE

    Fernández, Santiago

    2006-01-01

    Estamos en la Francia del siglo XVII. Su sociedad gira en torno a una vigorosa monarquía. Allí viven grandes pensadores y literatos: Descartes, Fermat, Pascal, Moliere, Racine, etc. Los juegos de dados, cartas y tableros con fichas son los entretenimientos más frecuentes. Pero, los juegos, cada vez más complicados, y las apuestas cada vez más elevadas crean la necesidad de calcular sus probabilidades de manera racional.

  14. Variabilidade genética de acessos de pitaya com diferentes níveis de produção por meio de marcadores RAPD Pitaya accesses genetic variability with different pruduction levels through RAPD markers

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    Keize Pereira Junqueira

    2010-09-01

    Full Text Available A espécie de pitaya mais cultivada atualmente é Hylocereus undatus, a pitaya-vermelha-de-polpa-branca. Colômbia e México são os principais produtores mundiais e, devido à sua rusticidade, a pitaya é considerada uma alternativa potencialmente viável também para o aproveitamento de solos pedregosos, arenosos e maciços rochosos. Apesar da crescente demanda, ainda não há uma cultivar lançada no mercado que atenda às necessidades climáticas de produção e às exigências do consumidor brasileiro. O presente trabalho é parte do programa de seleção e melhoramento da pitaya CPAC PY-01 da Embrapa Cerrados. Objetivou-se realizar o estudo da variabilidade genética de 16 acessos de pitayas mantidos na coleção de germoplasma da Embrapa Cerrados, apresentando diferentes características fenotípicas relacionadas especialmente à produção, por meio de marcadores moleculares RAPD (Random Amplified Polymorphic DNA. O DNA genômico de cada acesso foi extraído, e onze iniciadores decâmeros foram utilizados para a obtenção de marcadores moleculares RAPD, que foram convertidos em matriz de dados binários, a partir da qual foram estimadas as distâncias genéticas entre os acessos e realizadas análises de agrupamento e de dispersão gráfica. Foram obtidos 111 marcadores RAPD, perfazendo uma média de 10,1 marcadores por primer, dos quais 45 (40,54% foram polimórficos. As distâncias genéticas entre os 16 acessos variaram entre 0,006 e 0,148. As maiores distâncias genéticas foram obtidas entre os acessos "52" e "61", sendo que, em 2007, o primeiro produziu mais de 25 frutos, e o segundo, nenhum. Assim, deduz-se que, nesse caso, a próvável causa da variação seja genotípica. As menores distâncias genéticas foram constatadas entre os acessos "63"e "55" e entre "19"e "59". Os dois grupos apresentaram valores de produção próximos. Os marcadores moleculares RAPD mostraram que, mesmo dentro da mesma espécie, há variabilidade gen

  15. Post Kala-Azar dermal leishmaniasis following treatment with 20 mg/kg liposomal amphotericin B (Ambisome for primary visceral leishmaniasis in Bihar, India.

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    Sakib Burza

    Full Text Available BACKGROUND: The skin disorder Post Kala-Azar Dermal Leishmaniasis (PKDL occurs in up to 10% of patients treated for visceral leishmaniasis (VL in India. The pathogenesis of PKDL is not yet fully understood. Cases have been reported in India following therapy with most available treatments, but rarely in those treated with liposomal amphotericin B (Ambisome. Between July 2007 and August 2012 with the support of the Rajendra Memorial Research Institute (RMRI, Médecins Sans Frontières (MSF supported a VL treatment programme in Bihar, India-an area highly endemic for Leishmania donovani-in which 8749 patients received 20 mg/kg intravenous Ambisome as first-line treatment. This study describes the characteristics of patients who returned to the MSF supported treatment programme with PKDL. METHODS AND PRINCIPAL FINDINGS: Over a 5-year period, Ambisome was administered to 8749 patients with laboratory-confirmed VL (clinical signs, rK39 positive, with/without parasite confirmation in four intravenous doses of 5 mg/kg to a total of 20 mg/kg, with a high initial-cure rate (99.3% and low default rate (0.3%. All patients received health education highlighting the possibility and symptoms of developing PKDL, and advice to return to the MSF programme if these symptoms developed. This is an observational retrospective cohort study of the programme outcomes. Of the 8311 patients completing treatment for their first episode of VL, 24 (0.3% returned passively to the programme complaining of symptoms subsequently confirmed as PKDL, diagnosed from clinical history, appearance consistent with PKDL, and slit-skin smear examination. Of the 24 patients, 89% had macular lesions, with a median time (interquartile range to development of 1.2 (0.8-2.2 years following treatment. Comparison of the demographic and clinical characteristics of the VL patients treated with Ambisome who later developed PKDL, with those of the remaining cohort did not identify any significant

  16. L. (L. chagasi in aids and visceral leishmaniasis (kala-azar co-infection L. (L. chagasi em lesões cutâneas na co-infecção aids-calazar

    Directory of Open Access Journals (Sweden)

    Ana Maria Roselino

    2008-08-01

    Full Text Available Concomitant skin lesions in visceral leishmaniasis (VL or kala-azar are rare, being more common the description of post-kala-azar dermal leishmaniasis occurring post treatment of kala-azar. Skin lesions caused by Leishmania donovani are frequently seen in the aids-VL co-infection. In Brazil cutaneous or mucosal forms of tegumentary leishmaniasis concomitant with aids are more commonly registered. Here we present a case of aids-VL co-infection, with unusual cutaneous and digestive compromising attributed to L. (L. chagasi, with special attention to ecthymatous aspect of the lesion, allied to the absence of parasite on the histological skin biopsy.Lesões cutâneas, na vigência da leishmaniose visceral (LV ou calazar, são raramente observadas, sendo mais comum a ocorrência após o tratamento do calazar, conhecidas como lesões dérmicas pós calazar. Lesões cutâneas causadas por Leishmania donovani são freqüentemente observadas na co-infecção AIDS-LV. No Brasil, a concomitância das formas cutânea ou mucosa da leishmaniose tegumentar com a AIDS é mais comumente relatada. A seguir, relata-se um caso de co-infecção AIDS-LV com inusitado comprometimento digestivo e cutâneo, atribuído a L. (L. chagasi, chamando a atenção para o aspecto ectimatóide da lesão cutânea, aliado à ausência do parasito ao exame histopatológico da pele.

  17. Application of DNA (RAPD and ultrastructure to detect the effect of cadmium stress in Egyptian clover and Sudan grass plantlets

    Directory of Open Access Journals (Sweden)

    Amina A. Aly

    2012-04-01

    Full Text Available BackgroundIn recent years, several plant species have been used as bioindicators to evaluate the toxicity of environmental contaminants on vegetal organisms. In this study, Egyptian clover and Sudan grass seedlings were grown in four cadmium (Cd concentration levels (0.0, 25, 50 and 100 µM in MS media to analyze growth responses, Cd accumulation in the shoots and roots of plantlets, proline contents, chlorophylls content and MDA levels of both plantlets. As well as RAPD analysis and leaves ultrastructure were detected.ResultsThe results showed that there was a significant decrease in root and shoot lengths, Chl a, Chl b, total Chl and carotenoids contents for both Egyptian clover and Sudan grass. However, there was a significant increase in Cd accumulation, proline and malondialdehyde (MDA levels. The genetic variation between Egyptian clover and Sudan grass were evaluated using random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR markers to establish specific DNA markers associated with Cd stress. The results of transimssion electron microscopy (TEM showed a clear disorder in the Cd treated Egyptian clover and Sudan grass seedlings.ConclusionIn conclusion, biochemical, molecular and ultrastructure changes in Egyptian clover and Sudan grass could be used as a useful biomarker assay for the detection of genotoxic effects of Cd stress on plants. However, it is necessary to be further confirmed and optimized in the future research.

  18. Genetic diversity and relationship between cultivated clones of Dalbergia sissoo of wide geographical origin using RAPD markers

    Institute of Scientific and Technical Information of China (English)

    H. S. Ginwal; S. S. Maurya; P. Chauhan

    2011-01-01

    Random Amplified Polymorphic DNA (RAPD) polymorphism was employed to assess the genetic diversity in the elite germplasm of Dalbergia sissoo.Sixty-seven clones that are under cultivation in northern India,originated from six different states of India and Nepal were analyzed with 30 RAPD primers that generated a total of 342 fragments out of which 290 were polymorphic.Total genetic diversity (Ht) varied between 0.01 and 0.37,with an average of 0.19.Shannon's Information index (I) varied between 0.02 and 0.54,with an average of 0.31.Marker attributes like Polymorphism Information Content (PIC),Marker Index (MI) and Effective Multiplex Ratio (EMR) values were calculated to assess the discriminatory power of 30 primers used.The PIC values ranged from 0.01 to 0.37 with an average of 0.17 per primer and the EMR ranged from 0.17 to 21.00 with a mean of 8.66 across all genotypes.Closely related clones were CA9 and C51 with similarity index of 0.86 while the least similar or most dissimilar clones were C14 and S-DB showing similarity index of 0.58.The UPGMA-phenogram categorized the 67 clones into six clusters based on genetic similarity and dissimilarity.The clustering of clones in relation to their geographical location has been discussed.

  19. Authentication of Cordyceps sinensis by DNA Analyses: Comparison of ITS Sequence Analysis and RAPD-Derived Molecular Markers

    Directory of Open Access Journals (Sweden)

    Kelly Y. C. Lam

    2015-12-01

    Full Text Available Cordyceps sinensis is an endoparasitic fungus widely used as a tonic and medicinal food in the practice of traditional Chinese medicine (TCM. In historical usage, Cordyceps specifically is referring to the species of C. sinensis. However, a number of closely related species are named themselves as Cordyceps, and they are sold commonly as C. sinensis. The substitutes and adulterants of C. sinensis are often introduced either intentionally or accidentally in the herbal market, which seriously affects the therapeutic effects or even leads to life-threatening poisoning. Here, we aim to identify Cordyceps by DNA sequencing technology. Two different DNA-based approaches were compared. The internal transcribed spacer (ITS sequences and the random amplified polymorphic DNA (RAPD-sequence characterized amplified region (SCAR were developed here to authenticate different species of Cordyceps. Both approaches generally enabled discrimination of C. sinensis from others. The application of the two methods, supporting each other, increases the security of identification. For better reproducibility and faster analysis, the SCAR markers derived from the RAPD results provide a new method for quick authentication of Cordyceps.

  20. Authentication of Cordyceps sinensis by DNA Analyses: Comparison of ITS Sequence Analysis and RAPD-Derived Molecular Markers.

    Science.gov (United States)

    Lam, Kelly Y C; Chan, Gallant K L; Xin, Gui-Zhong; Xu, Hong; Ku, Chuen-Fai; Chen, Jian-Ping; Yao, Ping; Lin, Huang-Quan; Dong, Tina T X; Tsim, Karl W K

    2015-12-15

    Cordyceps sinensis is an endoparasitic fungus widely used as a tonic and medicinal food in the practice of traditional Chinese medicine (TCM). In historical usage, Cordyceps specifically is referring to the species of C. sinensis. However, a number of closely related species are named themselves as Cordyceps, and they are sold commonly as C. sinensis. The substitutes and adulterants of C. sinensis are often introduced either intentionally or accidentally in the herbal market, which seriously affects the therapeutic effects or even leads to life-threatening poisoning. Here, we aim to identify Cordyceps by DNA sequencing technology. Two different DNA-based approaches were compared. The internal transcribed spacer (ITS) sequences and the random amplified polymorphic DNA (RAPD)-sequence characterized amplified region (SCAR) were developed here to authenticate different species of Cordyceps. Both approaches generally enabled discrimination of C. sinensis from others. The application of the two methods, supporting each other, increases the security of identification. For better reproducibility and faster analysis, the SCAR markers derived from the RAPD results provide a new method for quick authentication of Cordyceps.

  1. Genetic Variation of the First Generation of Rodent Tuber (Typhonium flagelliforme Lodd. Mutants Based on RAPD Molecular Markers

    Directory of Open Access Journals (Sweden)

    Nesti Fronika Sianipar

    2015-04-01

    Full Text Available Rodent tuber (Typhonium flagelliforme Lodd. is a herbal plant from the Araceae family. The plant has high medical potential and is effective to cure cancer. However, the low level of its genetic variation limits its exploration for desirable traits. The low level of genetic variation in Rodent tuber is mainly due to its asexual reproduction system. It usually reproduces vegetatively via tuber separation. Therefore, gamma irradiation had been applied to rodent tuber in vitro to increase its genetic diversity. The objective of this study was to analyze the genetic diversity of the first generation (MV1 of gamma irradiated rodent tuber mutant using random amplified polymorphic DNA (RAPD markers. A total of 14 mutant DNA samples were analyzed with 14 RAPD primers. The result showed that 67 out of 123 DNA bands were polymorphic among mutant lines. Based on cluster analysis these mutants showed 0.78-0.97 genetic similarity. Cutting of dendogram at genetic distance of 0.89 produced four main clusters. Mutants with high genetic variation are now available. This increases the opportunity of obtaining mutant lines with high anti-cancer activity.

  2. Molecular and functional diversity of PGPR fluorescent Pseudomonads based on 16S rDNA-RFLP and RAPD markers.

    Science.gov (United States)

    Singh, Bhim Pratap

    2015-09-01

    The genetic and functional diversity of plant growth promoting rhizobacterial (PGPR) fluorescent pseudomonads associated with chickpea (Cicer arietinum L.) rhizosphere was analyzed. In total, 34 isolates along with two reference isolates were screened for various plant growth promoting traits (phosphorous solubilization, ACC deaminase, HCN, IAA and siderophore productions) and antagonist activity against four fungal phytopathogens and three bacterial pathogens. Most of the isolates, that showed PGPR activity, also showed antagonistic activity against all the three fungal pathogens. The genetic relationship was assessed by using random amplification of polymorphic DNA (RAPD) and PCR-restriction fragment length polymorphism (16S rDNA-RFLP). Relationship between both the markers was analyzed based on mantel test and a negative correlation was observed. The study concluded that PGPR traits appeared to be strain specific rather than specific to any phylogenetic group. The study also reported that 16S rDNA based profiling differentiated PGPR fluorescent Pseudomonas on the basis of location rather than biological trait. RAPD profiling could be useful to differentiate among the closely related isolates. The genetic and functional diversity of fluorescent pseudomonads, associated with the chickpea rhizosphere, has useful ecological role and potential utilization in sustainable agriculture.

  3. [Variability and phylogenetic relationships of the Cucumis sativus L. species inferred from NBS profiling and RAPD analysis].

    Science.gov (United States)

    Goriunova, S V; Gashkova, I V; Kosareva, G A

    2011-08-01

    Genetic variability of the Cucumis sativus species and its phylogenetic relationsips with other species of the genus were studied on the basis of RAPD marking and analysis of intra- and interspecific polymorphism of the nucleotide sequences of the NBS-LRR gene family in species of the genus Cucumis with the use of the NBS profiling method. According to RAPD analysis, cucumber cultivars from different geographic regions are highly similar, except for accessions k-3835 and k-3833 from Afghanistan. NBS profiling analysis revealed phylogenetically most distinct accessions expected to be characterized by specificity of resistance: k-3845 from Uzbekistan, k-3851 from Kyrgyzstan, line 701, k-3835 and k-3833 from Afghanistan, k-2757 and k-3079 from Netherlands, vr.k. 908 from Canada, k-2926 from Bulgaria, Russian cultivars Monastyrskii, Izyashchnyi, and Lel'. Three essentially different groups of species were distinguished, and the C. sativus species (subgenus Cucumis) was found to be distant from the species belonging to the subgenus Melo.

  4. Assessment of Genetic Stability Among In Vitro Plants of Arachis retusa Using RAPD and AFLP Markers for Germplasm Preservation

    Institute of Scientific and Technical Information of China (English)

    Rachel Fatima Gagliardi; Luiz Ricardo Hanai; Georgia Pacheco; Carlos Alberto Oliveira; Leonardo Alves Carneiro; José Francisco Montenegro Valls; Elisabeth Mansur; Maria Lucia Carneiro Vieira

    2007-01-01

    Arachis retusa Krapov. et W. C. Gregory et Valls is endemic in the West-central region of Brazil, occurring in areas endangered by human actions. The establishment of in vitro preservation methods for wild species of Arachis isan alternative to seed banks for germplasm storage, multiplication and distribution. The risk of genetic changesinduced by tissue culture and the monitoring of the genetic stability of the biological material before, during andafter storage must be considered in the context of conservation. Random amplified polymorphic DNA (RAPD) andamplified fragment length polymorphism (AFLP) fingerprinting were used to evaluate the genetic stability of invitro plants originated from cotyledons and embryo axes of A. retusa. Cotyledons originated shoots through directorganogenesis and embryo axes displayed multishoot formation induced by 110 mmol/L and 8.8 mmol/L BAP,respectively. Ninety genomlc regions (loci) generated from RAPD and 372 from AFLP analyses were evaluated. Allamplified fragments detected by both techniques in plants derived from the two explant types were monomorphic.The results indicate that the recovered shoots are genetically stable at the assessed genomic regions.

  5. Inheritance of Resistance to SMV3 and Identification of RAPD Marker Linked to the Resistant Gene in Soybean

    Institute of Scientific and Technical Information of China (English)

    ZHENG Cui-ming; CHANG Ru-zhen; QIU Li-juan

    2002-01-01

    One SMV resistant soybean line (95-5383) was crossed with four susceptible soybean varieties/line ( HB1, Tiefeng21, Amsoy, Williams) and one resistant introduced line PI486355. Their F1 and F2individuals were identified for SMV resistance by inoculation with SMV3. The results showed that in the four crosses of resistant × susceptible, F1 were susceptible and the ratio of F2 populations was 1 resistant : 3susceptible (mosaic and necrosis), indicating that 95-5383 carries one recessive gene that confer resistance to SMV3. There is segregation of susceptibility in F2 progenies from the cross of 95-5383 × PI486355, indicating that the SMV3 resistant gene in 95-5383 is located at different locus from PI486355. By bulked segregating analysis (BSA) in F2 populations of 95-5383 × HB1, one codominant RAPD marker OPN11980/1070 closely linked to SMV3 resistance gene amplified with RAPD primer OPN11 was identified. The DNA fragment OPN11980 was amplified in resistant parent 95-5383 and resistant bulk, and OPN111070 was amplified in susceptible parent HB1 and susceptible bulk. OPN11980/1070 was amplified in F1. Identification of the markers in F2 plants showed that the codominant marker OPN11980/1070 is closely linked to the SMV resistance locus in95-5383, with genetic distance of 2.1cM.

  6. Using cytogenetic analysis RAPD in determination of genetic variations among four species of ornamental fishes of family: Poecilidae (Order: Cyprinodontiform

    Directory of Open Access Journals (Sweden)

    Abu-Almaaty A.H.

    2015-01-01

    Full Text Available The karyological and molecular analysis of four fresh water fish species of Family: Poecilidae and their genetic relationships have been studied. Xiphophorus maculates and Xiphophorus hellerii have the same diploid chromosome number 2n=48, but they were different in their karyotypes. Poecilia sphenops and Poecilia reticulata have the same diploid chromosome number 2n=46 and the same fundamental number FN=46, also the same karyotype one group of acrocentric chromosomes. Nine RAPD primers, showed monomorphic bands, were used for the construction of the dendrogram and a similarity matrix. A total of 65 bands were obtained; 39 of them were monomorphic bands. Similarity values among the studied samples ranged from 21% to 38%. High similarity value was obtained between Xiphophorus maculates and Xiphophorus hellerii. (38% and the low similarity values were obtained between Xiphophorus hellerii and Poecilia reticulata (21%. The cluster analysis clearly differentiated Xiphophorus maculates and Xiphophorus hellerii from Poecilia sphenops and Poecilia reticulata. RAPD analysis confirmed that the four species under study are genetically different from each other. These cytogenetic data obtained can be applied for further studies in cytotaxonomy and evolutionary relationships of fishes.

  7. Efecto del peróxido de hidrógeno al 35% con y sin la activación de lámpara de diodos en la microdureza del esmalte

    OpenAIRE

    2013-01-01

    Objetivo. Comparar el efecto del clareamiento dental con peróxido de hidrógeno al 35% con y sin la activación de lámpara de diodos en la microdureza del esmalte dental. Material y métodos. Investigación experimental, longitudinal realizada con una muestra no probabilística, intencional de 30 premolares permanentes distribuidos al azar en dos grupos: grupo control (n=15) tratados con peróxido de hidrógeno al 35% y grupo experimental (n=15) tratados con peróxido de hidrógeno al 35% más...

  8. Identification of SSR and RAPD markers linked to a resistance allele for angular leaf spot in the common bean (Phaseolus vulgaris line ESAL 550

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    Gilvan Ferreira da Silva

    2003-12-01

    Full Text Available The objective of this study was to identify RAPD and SSR markers associated with a resistant allele for angular leaf spot (Phaeoisariopsis griseola from the line 'ESAL 550', derived from the Andean 'Jalo EEP 558' cultivar, to assist selection of resistant genotypes. The resistant line 'ESAL 550' and the susceptible cultivar 'Carioca MG' were crossed to generate F1 and F2 populations. One hundred and twenty F2:3 families were evaluated. The DNA of the 12 most resistant families was bulked and the same was done with the DNA of the 10 most susceptible, generating two contrasting bulks. One RAPD and one SSR marker was found to be linked in coupling phase to the resistant allele. The SSR marker was amplified by the primer PV-atct001(282C, and its distance from the resistant allele was 7.6 cM. This is the most useful marker for indirect selection of resistant plants in segregating populations. The RAPD marker was amplified by the primer OPP07(857C linked in coupling phase to the resistant allele, and distant 24.4 cM. Therefore, this RAPD marker is not so useful in assisting selection because it is too far from the resistant allele.

  9. Conversion of chromosome-specific RAPDs into SCAR-based anchor markers for onion linkage maps and its application to genetic analyses inother Allium species

    NARCIS (Netherlands)

    Masuzaki, S.; Miyazaki, T.; McCallum, J.; Heusden, van A.W.; Kik, C.; Yamashita, K.; Tashiro, Y.

    2008-01-01

    Integration of previously developed Allium cepa linkage maps requires the availability of anchor markers for each of the eight chromosomes of shallot (A. cepa L. common group Aggregatum). To this end, eight RAPD markers originating from our previous research were converted into SCAR markers via clon

  10. 酒酒球菌抗酒精基因RAPD标记的研究%RAPD specific molecular markers linked to alcohol resistance gene in Oenococcus oeni

    Institute of Scientific and Technical Information of China (English)

    王坐京; 李西柱; 周利国; 刘树文

    2009-01-01

    以抗酒精酒酒球菌和酒精敏性酒酒球菌为材料,提取基因组DNA构建抗性与敏性基因池,对RAPD-PCR反应体系进行优化,利用RAPD分子标记技术对45条随机引物进行筛选.结果引物S90在750bp附近扩增出了差异条带,且重复性好,这表明S90750是与酒酒球菌抗酒精基因相连锁的RAPD标记.%The study was carried out using the alcohol resistance Oenococcus oeni and the alcohol sensitive O.oeni as the material. After extracting Genomic DNA to construction resistance and the sensitive gene pools and optimizing the RAPD-PCR conditions, RAPD molecular marker technology was applied to screen in 45 random primers which were used to amplify the two types of gene pools. Only primer S90 was identified to be linked to the resistant gene nearby 750 bp, and the repeatability was good. Therefore, the RAPD markers S90750 are linked to the alcohol resistance gene in O.oeni.

  11. Characterization of Grain Amaranth (Amaranthus spp. Germplasm in South West Nigeria Using Morphological, Nutritional, and Random Amplified Polymorphic DNA (RAPD Analysis

    Directory of Open Access Journals (Sweden)

    Pamela E. Akin-Idowu

    2016-01-01

    Full Text Available Efficient utilization of plant genetic resources for nutrition and crop improvement requires systematic understanding of the important traits. Amaranthus species are distributed worldwide with an interesting diversity of landraces and cultivars whose leaves and seeds are consumed. Despite their potential to enhance food security and economic livelihoods, grain amaranth breeding to improve nutritional quality and adoption by farmers in sub-Saharan Africa is scanty. This study assessed the variation among 29 grain amaranth accessions using 27 phenotypic (10 morphological and 17 nutritional characters and 16 random amplified polymorphic DNA (RAPD primers. Multivariate analysis of phenotypic characters showed the first four principal components contributing 57.53% of observed variability, while cluster analysis yielded five groups at 87.5% similarity coefficient. RAPD primers generated a total of 193 amplicons with an average of 12.06 amplicons per primer, 81% of which were polymorphic. Genetic similarities based on Jaccard’s coefficient ranged from 0.61 to 0.88. The RAPD-based unweighted pair group method with arithmetic mean dendrogram grouped the accessions into nine clusters, with the same species clustering together. RAPD primers distinguished the accessions more effectively than phenotypic markers. Accessions in the different clusters as obtained can be exploited for heterotic gain in desired nutritional traits.

  12. Identification of Bacillus Probiotics Isolated from Soil Rhizosphere Using 16S rRNA, recA, rpoB Gene Sequencing and RAPD-PCR.

    Science.gov (United States)

    Mohkam, Milad; Nezafat, Navid; Berenjian, Aydin; Mobasher, Mohammad Ali; Ghasemi, Younes

    2016-03-01

    Some Bacillus species, especially Bacillus subtilis and Bacillus pumilus groups, have highly similar 16S rRNA gene sequences, which are hard to identify based on 16S rDNA sequence analysis. To conquer this drawback, rpoB, recA sequence analysis along with randomly amplified polymorphic (RAPD) fingerprinting was examined as an alternative method for differentiating Bacillus species. The 16S rRNA, rpoB and recA genes were amplified via a polymerase chain reaction using their specific primers. The resulted PCR amplicons were sequenced, and phylogenetic analysis was employed by MEGA 6 software. Identification based on 16S rRNA gene sequencing was underpinned by rpoB and recA gene sequencing as well as RAPD-PCR technique. Subsequently, concatenation and phylogenetic analysis showed that extent of diversity and similarity were better obtained by rpoB and recA primers, which are also reinforced by RAPD-PCR methods. However, in one case, these approaches failed to identify one isolate, which in combination with the phenotypical method offsets this issue. Overall, RAPD fingerprinting, rpoB and recA along with concatenated genes sequence analysis discriminated closely related Bacillus species, which highlights the significance of the multigenic method in more precisely distinguishing Bacillus strains. This research emphasizes the benefit of RAPD fingerprinting, rpoB and recA sequence analysis superior to 16S rRNA gene sequence analysis for suitable and effective identification of Bacillus species as recommended for probiotic products.

  13. Molecular Characterization and Genetic Diversity Analysis of Sweet Orange Citrus sinensis L. Osbeck Cultivars in Iraq Using RAPD Markers

    Directory of Open Access Journals (Sweden)

    Ali Saeed Atiyah AL-Janabi

    2016-03-01

    Full Text Available Sweet orange (Citrus sinensis L. Osbeck is one of the most important commercially cultivated fruit crops of Citrus. Genetic diversity and inter-relationship among 5 cultivars (Indian, Iraqi, Japanese, Syrian, Egyptian of C. sinensis were analyzed based on RAPD markers. Six primers generated reproducible and easily storable RAPD profiles with a number of amplified DNA fragments ranging from 6 to 14 fragment bands. The total number of amplicons detected was 51, including 14 fragments unique bands with average reached 2.8 fragments ̸ primers. While the number of polymorphic ranged from 0 to 8 with an average reached 4.4 fragments ̸ primers with the polymorphic percentage ranged from 0% to 57.1%. While the number of monomorphic ranged from 2 to 5 fragment bands and was total of the monomorphic 15 fragments with an average reached 3 fragments ̸ primers with the monomorphic percentage was 14.2 % to 83.3%. A maximum numbers of amplicons was amplified with primer OPS-238 reached 14 fragments while the minimum number of fragments was amplified with primer OPS-253 reached 6 fragments. The highest number of polymorphic bands reached 8 fragments was obtained with primer OPS-238 with high percentage 57.1%, while the highest number of monomorphic bands reached 5 fragments with high percentage 83.3% was obtained with primer OPS-253. RAPD markers detected genetic distance and similarity, amaximum genetic distance value was observed between Japanese (Jap and Syrian (Syr cultivars reached 0.530 with less similarity value reached 47%, a minimum genetic distance value was observed between sweet Iraqi (Irq and Indian (Ind cultivars reached 0.239 with high similarity value reached 76.1%. The similarity matrices were employed in the cluster analysis to generate a dendrogram using the UPGMA method. The cluster tree analysis showed that the sweet orange cultivars were broadly divided into two main groups A and B with similarity reached 50%. A group including

  14. Establishment of Nanfeng citrus genomic DNA's PCR- RAPD optimal conditions%南丰蜜桔基因组DNA RAPD-PCR最佳反应体系的建立

    Institute of Scientific and Technical Information of China (English)

    郑敏; 罗玉萍; 李思光; 朱必凤

    2001-01-01

    以南丰蜜桔树叶为基因组DNA提取材料,建立了其RAPD-PCR分析的最佳反应体系:模板DNA1.5ng/μl,随机引物0.6μM@L-1,dNTPs各0.1mM@L-1,MgCl2.0mM@L-1,BSA 250ng/μl,Tris-HCl 10mM@L-1,pH9.0,KCl 50mM@L-1,Nonidet P400.1%,Taq酶0.5u,总反应体积20μl.

  15. Análise da variabilidade genética de arnica (Lychnophora ericoides Less. - Asteraceae usando marcadores RAPDs Genetic variability analysis of arnica (Lychnophora ericoides Less. - Asteraceae using RAPD markers

    Directory of Open Access Journals (Sweden)

    Luciana Queiroz Melo

    2009-03-01

    Full Text Available O objetivo deste trabalho foi analisar e quantificar a variabilidade genética entre e dentro das populações de arnica por meio de marcadores RAPD. Foram amostradas quatro populações na região geoeconômica do Distrito Federal: Parque Nacional de Brasília (2, Fazenda Água Limpa - UnB (1 e Reserva do Instituto Brasileiro de Geografia e Estatística (IBGE (1. Folhas de 24 indivíduos de cada região foram coletadas, totalizando 96 indivíduos. Num total de 105 iniciadores testados foram selecionados 15, totalizando 60 bandas polimórficas. Marcadores RAPDs selecionados foram analisados com a utilização dos programas NTSYS e Amova. O dendrograma obtido pelo método UPGMA e coeficiente de dissimilaridade Dice evidenciou quatro agrupamentos consistentes, com índice de dissimilaridade variando entre 62 a 71%. O teste de Mantel aplicado estabeleceu uma correlação cofenética com valores de r = 0.82, significando que as distâncias geográficas entre as populações amostradas estão correlacionadas com a distância genética. A análise de AMOVA mostrou uma percentagem variabilidade genética entre populações de 35,7% e dentro de populações de 64,3%, evidenciando uma alta variação entre populações, sendo um importante resultado para definição de uma estratégia de conservação da espécie que se encontra em situação vulnerável à extinção.The main objective of this research was to analyze and quantify the genetic variability within and between populations of arnica using RAPD markers. Four populations from Federal District area, Brazil were sampled: Parque Nacional de Brasília - (2 , Fazenda Água Limpa -UnB (1, and Reserva do Instituto Brasileiro de Geografia e Estatística (IBGE (1. Leaves from twenty-four individuals from each population were collected and preserved under refrigeration. Fifteen primers were selected from 105 tested, totalizing 60 polymorphic bands. Scored RAPD markers were analyzed using NTSYS and Amova

  16. Differentiation of Candida species obtained from nosocomial candidemia using RAPD-PCR technique Diferenciação de espécies de Candida obtidas de candidemia nosocomial pela técnica de RAPD-PCR

    Directory of Open Access Journals (Sweden)

    Henrique Maia Valério

    2006-04-01

    Full Text Available Thirteen strains of the genus Candida were isolated from catheter, urine and surgical wounds from individual patients of the Santa Casa de Misericórdia, Belo Horizonte, MG, Brazil. Ten strains were characterized as Candida albicans, two as Candida glabrata, and one as Candida parapsilosis. Isolates were evaluated for molecular relatedness by random amplified polymorphic DNA technique using 15 primers. The analysis of the genomic DNA obtained revealed a low intraspecific polymorphism and did not allow for the differentiation between strains of the same species obtained from distinct clinical sources (catheter, urine and surgical wounds. The RAPD profiles generated were able to differentiate among the species of Candida albicans, Candida parapsilosis and Candida glabrata strains isolated in this study.Treze amostras de leveduras do gênero Candida foram isoladas de catéter, urina e feridas cirúrgicas de pacientes da Santa Casa de Misericórida de Belo Horizonte, MG, Brasil. Dez amostras foram identificadas como Candida albicans, duas como Candida glabrata e uma como Candida parapsilosis. Os isolados foram avaliados quanto ao perfil molecular pela técnica de amplificação aleatória de DNA polimórfico utilizando 15 iniciadores. A análise do DNA genômico obtido revelou um baixo polimorfismo intraespecífico e não permitiu a diferenciação entre amostras da mesma espécie obtidas a partir de diferentes espécimes clínicos (catéter, urina e feridas cirúrgicas. Os perfis de RAPD obtidos foram capazes de diferenciar entre as espécies Candida albicans, Candida parapsilosis e Candida glabrata isoladas neste estudo.

  17. Estimativa da similaridade genética e identificação de cultivares do morangueiro por análise de RAPD Estimate of the genetic similarity and identification of strawberry cultivars by RAPD analysis

    Directory of Open Access Journals (Sweden)

    José Henrique Conti

    2002-06-01

    Full Text Available Caracteres moleculares do morango foram avaliados para conhecer cultivares que estão sendo introduzidas no Brasil. Utilizou-se o método do polimorfismo de DNA amplificado ao acaso (RAPD. Os caracteres moleculares com maior poder de discriminação foram os "marcadores" gerados pelos "primers" Operon B8, Operon B19 e Operon G5, que foram eficientes para discriminar as vinte e seis cultivares estudadas. Os dados foram interpretados com o auxílio de dendograma, mapa de bandas, quadro de identificação e chave dicotômica. Foi possível distinguir seis grupos de similaridade, dois deles com cultivares selecionadas no Brasil, sendo um com 'Campinas', 'Agf 80', 'Piedade', 'Jundiaí' e 'Monte Alegre' e o outro com 'Obaira' e 'Mantiqueira'; três grupos com cultivares introduzidas, sendo o primeiro com 'Lassen', 'Reiko', 'Chandler', 'Pajaro', 'Blackmore' e 'Seascape', o segundo com 'Fern' e 'Oso Grande' e o terceiro com 'Florida Belle' e 'Selva' O último grupo reuniu as cultivares 'Dover' e 'Dabreak' junto com 'Princesa Isabel'.Strawberry cultivars introduced in Brazil were identified through molecular study. The method of Random Amplified Polymorphic DNA (RAPD was used. Molecular characters with larger discrimination power were produced by the primers Operon B8, Operon B19 and Operon G5. These were efficient to discriminate the twenty six studied cultivars. Data were analised through dendogram, bandmap, picture and dicotomic key. Six similarity groups were distinguished: Two with cultivars selected in Brazil, one with 'Campinas', 'Agf 80', 'Piedade', 'Jundiaí' and 'Monte Alegre' and the other with 'Obaira' and 'Mantiqueira'; three with introduced cultivars the first of which 'Lassen', 'Reiko', 'Chandler', 'Pajaro', 'Blackmore' and 'Seascape', the second with 'Fern' and 'Oso Grande' and the third with 'Florida Belle' and 'Selva'. The last group united the cultivars 'Dover' and 'Dabreak' with 'Princesa Isabel'.

  18. Similaridade genética de populações naturais de pimenta-de-macaco por análise RAPD Genetic similarity of natural populations of pimenta-de-macaco (Piper aduncum L. obtained throug RAPD analysis

    Directory of Open Access Journals (Sweden)

    José Maria D. Gaia

    2004-12-01

    Full Text Available A espécie conhecida como pimenta-de-macaco (Piper aduncum L. possui grande potencial para exploração econômica em função da comprovada utilidade do seu óleo essencial na agricultura e saúde humana. O objetivo deste trabalho foi caracterizar a diversidade genética de populações naturais dessa planta. Um total de dezoito acessos da planta, provenientes de quatro procedências da Amazônia Brasileira, foi examinado por meio de locos de DNA, gerados por análise RAPD (polimorfismo de DNA amplificado ao acaso. O estudo evidenciou a existência de real diversidade entre as populações examinadas, sendo provável que dentro das localidades investigadas, os padrões da diversidade genética acompanhem os padrões de distribuição geográfica.The species known as pimenta-de-macaco (Piper aduncum L. has great economic explotation potential based on the proved usefulness of the essential oil in agriculture and human health. The genetic diversity of their natural populations was characterized. A total of eighteen accessions of the plant, obtained from four different origins in the Brazilian Amazon, was examined by means of DNA loci, generated by RAPD analysis. Real genetic diversity was observed between the analyzed populations and it appear that the patterns of the genetic diversity follow the patterns of the geographical distribution.

  19. DIVERGÊNCIA GENÉTICA ENTRE DOZE GENÓTIPOS DE ABACAXIZEIRO (Ananas comosus L, Merril. ESTIMADA POR ANÁLISE DE MARCADORES RAPD GENETIC DIVERGENCE AMONG TWELVE GENOTYPES OF PINEAPLE (Ananas comosus L, Merril. ESTIMATED BY RAPD MARKERS

    Directory of Open Access Journals (Sweden)

    CÍCERO DONIZETE PEREIRA

    2001-08-01

    Full Text Available Por meio de estudos moleculares, este trabalho determinou a distância genética entre 12 genótipos de A. comosus por marcadores RAPD (Random Amplified Polymorphic DNA, utilizando 11 "primers" decâmeros da OPERON Technologies Inc. Dos 12 genótipos , 1 foi proveniente da Jamaica, 2 do Estado do Acre (Quinari e RBR-1, 2 do Estado do Maranhão (Turiaçu e São Domingos, 3 do Estado do Piauí (Cefas, Floriano-1 e Floriano-2, 2 do Estado da Bahia (Monte Alegre-1 e Monte Alegre-2 e 2 de Minas Gerais (Pérola e Smouth Cayenne. Pela análise de "cluster", utilizando o método de UPGMA, foi constatada uma grande divergência entre os genótipos de A. comosus estudados com a separação destes em dois grupos a uma distância genética de 31,1%.Through molecular studies, the genetic distance among 12 genotypes of A. comosus for RAPD markers (Random Amplified Polymorphic DNA was determined using 11 primers (decamers of the OPERON Technologies Inc. From the 12 genotypes, 1 came from Jamaica, 2 from the State of Acre (Quinari and RBR-1, 2 from the State of Maranhão (Turiaçu and São Domigos, 3 the State of Piauí (Cefas, Floriano-1 and Floriano-2, 2 from the State of Bahia (Monte Alegre-1 and Monte Alegre-2 and 2 from Minas Gerais (Pérola and Smooth Cayenne. By the analysis of cluster, using the method of UPGMA, a great divergence was verified among the genotypes of A. comosus studied or tima separation in two groups of a genetic distance of 31,1%.

  20. Determinación de la variabilidad genética entre aislamientos de Rosellinia sp. Rosellinia bunodes y Rosellinia pepo mediante la técnica de amplificación aleatoria de polimorfismos de DNA (RAPD y análisis de los espaciadores de transcritos internos (ITSS

    Directory of Open Access Journals (Sweden)

    Ángel Díaz Jorge Evelio

    2004-12-01

    Full Text Available Teniendo en cuenta que las medidas preventivas son las más eficientes para reducir la incidencia de muchas enfermedades, la implementación de estrategias que permitan el diagnóstico temprano y oportuno de fitopatógenos de cultivos, en especial aquellos de tardío rendimiento, es el objetivo principal de este trabajo. Los hongos fitopatógenos del género Rosellinia se consideran habitantes naturales del suelo. Sin embargo, su fase parasítica se asocia a la aparición de enfermedades radiculares principalmente en cultivos de café, papa y cacao, que generalmente se
    relaciona con el aumento en la oferta de materia orgánica, lo cual beneficia el crecimiento sus poblaciones y las de otros microorganismos que pueden ser patógenos. Con el fin de avanzar en el conocimiento de las especies Rosellinia sp., Rosellinia bunodes y Rosellinia pepo y de desarrollar estrategias de diagnóstico para evitar las pudriciones radiculares que estas ocasionan en cultivos de importancia agroeconómica y forestal, se utilizaron dos metodologías moleculares que permiten
    realizar un primer acercamiento a la variabilidad genética que presentan diferentes aislamientos de estas especies. Mediante el análisis de RAPD (Amplificación Aleatoria de Polimorfismos de ADN y la secuenciación de las regiones de rDNA-ITS (Espaciadores de Transcritos Internos fue posible establecer un alto grado de variabilidad entre las cepas, aún dentro de una misma especie. Dichas metodologías están basadas en la reacción en cadena de la polimerasa (PCR que ha sido extensamente utilizada tanto en la identificación de un organismo por medio de la amplificación selectiva de su ADN, como al ser aplicada para explorar la variabilidad que posee un genoma. Las cepas de Rosellinia sp. presentaron gran variación en los perfiles electroforéticos de RAPD constituyendo dos ramas aisladas en el dendrograma de similaridad construido a partir de matrices binarias. De la misma manera

  1. Diversidade genética de pitayas nativas do cerrado com base em marcadores RAPD Genetic diversity of native pitaya native from brazilian savannas with basis on RAPD markers

    Directory of Open Access Journals (Sweden)

    Keize Pereira Junqueira

    2010-09-01

    Full Text Available As pitayas do Cerrado vegetam naturalmente sobre maciços rochosos de arenito ou quartzito, troncos de árvores e em solos arenosos de campos rupestres de Minas Gerais, Bahia, Goiás, Distrito Federal, Tocantins, Rio de Janeiro e Bahia, havendo fortes evidências de que a região central do Brasil seja o maior centro de dispersão das pitayas, tendo em vista a grande diversidade fenotípica observada em acessos coletados. Objetivou-se realizar o estudo da diversidade genética de 13 acessos de pitayas mantidos na coleção de germoplasma da Embrapa Cerrados por meio de marcadores moleculares RAPD (Random Amplified Polymorphic DNA. O DNA genômico de cada acesso foi extraído, e quatorze iniciadores decâmeros foram utilizados para a obtenção de marcadores moleculares RAPD, que foram convertidos em matriz de dados binários, a partir da qual foram estimadas as distâncias genéticas entre os acessos com base no complemento do coeficiente de similaridade de Nei e Li (1979 e realizadas análises de agrupamento e de dispersão gráfica. Foram obtidos 162 marcadores RAPD, perfazendo uma média de 11,57 marcadores por primer. Do total de marcadores, 154 (95,06% foram polimórficos. As distâncias genéticas variaram entre 0,088 e 0,848, sendo que os maiores valores observados se referem a distância entre o acesso de Unaí-MG e o acesso Seleção Embrapa Cerrados. O acesso que mais se diferenciou dos demais foi "Unaí-MG", que apresentou uma distância genética média de 0,675 em relação aos demais acessos. A alta distância genética verificada é devido ao fato de os referidos acessos não pertencerem à mesma espécie. Os agrupamentos dos acessos de pitaya pouco se relacionaram com a origem geográfica dos mesmos. A grande diversidade genética das pitayas encontradas no Cerrado permite incluir esse Bioma no centro de diversidade e abre boas perspectivas para maiores estudos acerca do potencial dessa frutífera.Brazilian savanna pitayas

  2. Molecular analysis (RAPD-PCR) of inter-strain hybrids of the Paramecium aurelia species complex (Ciliophora, Protozoa).

    Science.gov (United States)

    Przyboś, Ewa; Prajer, Małgorzata; Greczek-Stachura, Magdalena

    2005-01-01

    RAPD-PCR analysis showed that species of the Paramecium aurelia complex possessed characteristic band patterns and that the majority were also polymorphic intra-specifically. A comparison of band patterns was performed for some inter-strain hybrids within P. primaurelia, P. tetraurelia, P. pentaurelia, P. septaurelia, P. octaurelia, P. decaurelia, P. dodecaurelia, P. tredecaurelia, and P. quadecaurelia to band patterns characteristic for the parental strains. The investigations, however, did not reveal a close correlation between the degree of inbreeding characteristic for the species and similarity of genotypes. A low similarity of hybrid and parental band patterns was observed in P. octaurelia, P. dodecaurelia, P. quadecaurelia and also P. primaurelia. A high similarity of band patterns of hybrid and parental strains was found in P. tetraurelia, P. septaurelia, P. decaurelia, and P. tredecaurelia.

  3. Molecular characterization of RAPD and SCAR marker linked to the frog-eye leaf spot resistance gene in soybean

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Two fragments SCS3620 and SCS3580 of the co-dominant marker OPS03620&580 that were linked to the resistance gene of soybean frog-eye leaf spot have been completely sequenced.A significant insertion of 30 bp is the main reason of the polymorphism between the two fragments.The results of Southern hybridization indicate that SCS3620 derives from a single- or low-copy sequence and can be used as an RFLP probe.A co-dominant SCAR marker SCS3620&580 has been developed based on the sequences.The segregation of SCS3620&580 is similar to that of RAPD marker OPS03620&580.Significant polymorphism has been shown between resistant and susceptible genotypes when 62 soybean genotypes were surveyed for the SCAR marker.Therefore,the marker can be used in the resistance breeding of soybean frog-eye leaf spot by marker-assisted selection.

  4. RAPD-based genetic analysis of offsprings from the sexual cross using allotetraploid citrus somatic hybrid as pollen parent

    Institute of Scientific and Technical Information of China (English)

    YI HuaLin; DENG XiuXin

    2007-01-01

    Huazhong Agricultural University, Wuhan 430070, ChinaThirty-one polymorphic decamer primers were selected to genotype 92 progenies from the cross between Yiben No,4, a monoembryonic diploid F1 hybrid of Citrus reticulata Blanco cv Huanongbendizao tangerine and C. ichangensis Swingle, and [Hamlin sweet orange + Rough lemon], an allotetraploid somatic hybrid of Citrus sinensis Osbeck cv. Hamlin and C. jambhiri Lush cv. Rough Lemon. x2(Chi-square) analysis of RAPD markers in the progenies indicated they were randomly transmitted from the four donor parents, without significant difference between the diploids and triploids. However,these progenies were clustered into three major groups using dendrogram constructed by UPGMA,skewed to three parents in certain degrees, 15 (13 triploids and 2 diploids) to Hamlin, 16 (9 and 7) to Yiben No. 4, and 61 (57 and 4) to [Hamlin sweet orange + Rough Lemon] from which genomic contribution was predominant in progenies, respectively.

  5. RAPD-based genetic analysis of offsprings from the sexual cross using allotetraploid citrus somatic hybrid as pollen parent

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Thirty-one polymorphic decamer primers were selected to genotype 92 progenies from the cross be- tween Yiben No.4, a monoembryonic diploid F1 hybrid of Citrus reticulata Blanco cv Huanongbendizao tangerine and C. ichangensis Swingle, and [Hamlin sweet orange + Rough lemon], an allotetraploid somatic hybrid of Citrus sinensis Osbeck cv. Hamlin and C. jambhiri Lush cv. Rough Lemon. χ2 (Chi-square) analysis of RAPD markers in the progenies indicated they were randomly transmitted from the four donor parents, without significant difference between the diploids and triploids. However, these progenies were clustered into three major groups using dendrogram constructed by UPGMA, skewed to three parents in certain degrees, 15 (13 triploids and 2 diploids) to Hamlin, 16 (9 and 7) to Yiben No. 4, and 61 (57 and 4) to [Hamlin sweet orange + Rough Lemon] from which genomic contribu- tion was predominant in progenies, respectively.

  6. MOLECULAR CHARACTERIZATION BY USING RANDOM AMPLIFIED POLYMORPHIC DNA (RAPD ANALYSIS OF SALMONELLA ENTERITIDIS ISOLATES RECOVERED FROM AVIAN AND HUMAN SOURCES

    Directory of Open Access Journals (Sweden)

    E. YAQOOB, I. HUSSAIN AND S. U. RAHMAN

    2007-04-01

    Full Text Available Random amplified polymorphic DNA (RAPD analysis was applied for molecular characterization of five Salmonella enteritidis strains from different avian sources and human cases of infection. A total of 16 primers were used and only five primers showed good discriminatory power for all five isolates. Dendrogram showed a common lineage among all five isolates. There was a close genetic relationship among isolates of eggs and human sources, while there was less pronounced homology among isolates of broiler meat and human sources. On the basis of results we have found that an endemic strain of S. enteritidis is prevalent between the poultry derived food and humans which gives us an insight to genetic diversity of S. enteritidis from these sources.

  7. 基于RAPD分析用的佛手DNA的提取%DNA extraction for RAPD analysis in Bergamot

    Institute of Scientific and Technical Information of China (English)

    马伯军; 陈镖; 章斌轶; 张东旭; 陈秉初

    2002-01-01

    对佛手DNA提取过程从不同方面进行了改良、优化,筛选出较理想的佛手DNA提取方法:每eppendorf管鲜样品取量为0.1 g~0.25 g,700 μL提取缓冲液,加提取液前用液氮研磨,Vc(抗坏血酸)添加量为{w(V-C/鲜样)}={0.1}.该方法分离出DNA的 A-{260}/A-{280}大于1.9,符合佛手RAPD(随机扩增多态DNA)分析要求,在佛手遗传多态性分析和品种分子鉴定中具有良好的应用前景.

  8. ANALYSIS OF EVOLUTIONARY RELATIONSHIP BETWEEN ASTASIA LONGA AND EUGLENA GRACILIS BY USING RAPD TECHNIQUE AND CLADISTIC ANALYSIS

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Although both Astasia longa and Euglena gracilis belong to different genera, they share many morphological characters except that A. longa has no chloroplast. In the 1940's, on the basis of the finding that in darkness or upon addition of some chemicals, E. gracilis would fade reversibly or irreversibly, some scholars hypothesised that A. longa evolved from E. gracilis by losing chloroplast. The authors' use of RAPD and cladistic analyses in a study on the evolutionary relationship between A .longa and E. gracilis showed that the A. longa 's relationship with E. gracilis was closer than that with other green euglenoids. This proves the hypothesis that A. longa evolved from E. gracilis is reasonable. The results of this study suggest that saprophytic colorless euglenoids were transformed from green euglenoids by losing their choroplasts.

  9. Analysis of evolutionary relationship between Astasia longa and Euglena gracilis by using RAPD Technique and cladistic analysis

    Science.gov (United States)

    Wang, Jiang-Xin; Shi, Zhi-Xin; Gan, Xiao-Ni; Xie, Shu-Lian

    2001-03-01

    Although both Astasia longa and Euglena gracilis belong to different genera, they share many morphological characters except that A. longa has no chloroplast. In the 1940's, on the basis of the finding that in darkness or upon addition of some chemicals, E. gracilis would fade reversibly or irreversibly, some scholars hypothesised that A. longa evolved from E. gracilis by losing chloroplast. The author's use of RAPD and cladistic analyses in a study on the evolutionary relationship between A. longa and E. gracilis showed that the A. longa's relationship with E. gracilis was closer than that with other green euglenoids. This proves the hypothesis that A. longa evolved from E. gracilis is reasonable. The results of this study suggest that saprophytic colorless euglenoids were transformed from green euglenoids by losing their choroplasts.

  10. Identification of a RAPD marker linked to the Co-6 anthracnose resistant gene in common bean cultivar AB 136

    Directory of Open Access Journals (Sweden)

    Alzate-Marin Ana Lilia

    2000-01-01

    Full Text Available The pathogenic variability of the fungus Colletotrichum lindemuthianum represents an obstacle for the creation of resistant common bean (Phaseolus vulgaris L. varieties. Gene pyramiding is an alternative strategy for the development of varieties with durable resistance. RAPD markers have been proposed as a means to facilitate pyramiding of resistance genes without the need for multiple inoculations of the pathogens. The main aims of this work were to define the inheritance pattern of resistance present in common bean cultivar AB 136 in segregating populations derived from crosses with cultivar Rudá (susceptible to most C. lindemuthianum races and to identify RAPD markers linked to anthracnose resistance. The two progenitors, populations F1 and F2, F2:3 families and backcross-derived plants were inoculated with race 89 of C. lindemuthianum under environmentally controlled greenhouse conditions. The results indicate that a single dominant gene, Co-6, controls common bean resistance to this race, giving a segregation ratio between resistant and susceptible plants of 3:1 in the F2, 1:0 in the backcrosses to AB 136 and 1:1 in the backcross to Rudá. The segregation ratio of F2:3 families derived from F2 resistant plants was 1:2 (homozygous to heterozygous resistant. Molecular marker analyses in the F2 population identified a DNA band of approximately 940 base pairs (OPAZ20(940, linked in coupling phase at 7.1 cM of the Co-6 gene. This marker is being used in our backcross breeding program to develop Rudá-derived common bean cultivars resistant to anthracnose and adapted to central Brazil.

  11. Genetic Diversity in Haploid Nicotiana alata Induced by Gamma Irradiation, Salt Tolerance and Detection of These Differences by RAPD

    Directory of Open Access Journals (Sweden)

    Ayman EL-FIKI

    2016-03-01

    Full Text Available Haploid plants of Nicotiana alata were cultured in vitro on MS medium with IAA + KIN. The resulting plantlets were irradiated using gamma radiation doses of 10, 15, 20 and 25 Gy. Single node pieces were cut and transferred onto fresh MS medium. Gamma radiation doses caused the death of 9% and up to 28% of explants. NaCl concentrations caused the death of 8% up to 36% of explants, while the combined effect between gamma radiation doses and salinity had an impact suffused on the percentage of survival. The combined effect of gamma radiation doses 20 Gy and 25 Gy on NaCl concentrations of 100, 150 and 200 mM were deadly. Even more, the combined effect of gamma radiation doses and salinity had a severe negative impact on both the proline content and total soluble protein. Random amplified polymorphic DNA (RAPD analysis was used to determine the degree of genetic variation in treated haploid Nicotiana alata plants. Total genomic DNAs from different haploid plantlets treated were amplified using five arbitrary primers. Two hundred and seventy bands were detected from plantlets irradiated with doses of 15, 20 and 25 Gy, with polymorphic band number 226 (83.7%. The total number of bands resulted from plant grew on 150 mM and 200 mM NaCl were 260 bands with polymorphic bands 185 (85.6%. However, the total number of bands produced from combined effects between gamma rays and salinity (20 Gy X 50 mM NaCl, 20 Gy X 100 mM NaCl and 25 Gy X 50 mM NaCl were 270, with polymorphic band number 231 (85.5%. High similarity between treatments was revealed. Treatments relationships were estimated through cluster analysis (UPGMA based on RAPD data.

  12. Studi Epidemiologi Agen Zoonosis Escherichia coli O157:H7 melalui Analisis Random Amplification of Polymorphic DNA (RAPD

    Directory of Open Access Journals (Sweden)

    I Wayan Suardana

    2012-11-01

    Full Text Available Epidemiological studies of zoonotic agent Escherichia coli O157:H7 have been analyzed pheneticallyand or phylogenetically. In a phenetic classification, micoorganisms are arranged into groups (phena onthe basis of high overall similarity using both phenotypic and genotypic methods without judgementaspect of its ancestry or evolutionary. Due to its importance to epidemiological aspect, the study of geneticvariation of isolates origin from some sources need to be conducted in order to trace the routes of infection.A total of 20 samples obtained from some sources i.e clinically human feces, non-clinically human feces,cattle feces, chicken feces, and beef feces were used in this study. The study was started by confirming allof the isolates using O157 latex agglutination test and H7 antiserum test, followed by genomic DNAanalysis by random amplification of polymorphic DNA /RAPD methods. RAPD results were analyzed using a simple matching coeficient (Ssm and alogorhythm unweighted pair group method using arithmeticaverages (UPGMA programe. Results showed there were range of genetic DNA from local isolates (75.1–99,6% which was almost similar to ATCC 43894 control isolate. The highest similarity (99.6% to ATCC43894 control was showed by SM-7(1 isolate obtained from cattle fecal and KL-68(1, isolate obtainedfrom clinically human fecal. In addition, KL-52(7 obtained from clinically human fecal had high similarity(99.6% to MK-35 isolate obtained from chicken fecal. On the other hand, DS-21(4 and DS-16(2 isolatesthat were obtained from beef had high similarity (84.9% to other isolates including ATCC 43894 controlisolate. The highest similarity of E. coli O157:H7 isolates that were obtained from cattle feces, beef, andchicken feces to human feces isolate indicated that there were both cattle and chicken were potentialreservoirs of the zoonotic agen which can be transmitted to human.

  13. Comparison between Oligoryzomys nigripes and O. flavescens by RAPD and genetic diversity in O. nigripes (Rodentia, Cricetidae

    Directory of Open Access Journals (Sweden)

    AJ Mossi

    Full Text Available The genus of Oligoryzomys includes species of small size, morphologically similar, which may impede taxonomic identification, mainly between O. flavescens (Waterhouse, 1837 and O. nigripes (Olfers, 1818. The main objective of this work was to investigate whether the RAPD markers are capable of genetically differentiating the specimens O. nigripes and O. flavescens, coming from Rio Grande do Sul (RS and Santa Catarina (SC states, and also to estimate the genetic variability among populations of O. nigripes, with the Uruguay River as a geographical barrier. For this purpose, samples were collected in fragments of forests situated in the North of RS, at FLONA (Floresta Nacional de Passo Fundo and in fragments from SC, close to the Uruguay River. The karyotyping of two samples for each species was carried out and compared using the RAPD technique together with non- karyotyped individuals. Samples of O. nigripes presented 2n = 62; NA = 82, with submetacentric arms on the largest chromosomes, while samples of O. flavescens showed 2n = 64; NA = 66, with the largest chromosomes presenting acrocentric morphology, making such a result the main difference between the species. The analysis was able to detect two distinct groups, being the first one with karyotyped O. flavescens and the second with karyotyped O. nigripes. Identification afforded 211 loci, among them 181 (85.78% polymorphic. The Jaccard similarity coefficient was in the range of 0.45 to 0.87. The UPGMA and Main Coordinate Analysis techniques demonstrated the existence of heterogeneous genetics among populations, but did not separate them completely in terms of geographical standards, and they are not influenced by the Uruguay River, which did not act as an efficient barrier.

  14. Micropropagation and validation of genetic and biochemical fidelity amongst regenerants of Cassia angustifolia Vahl employing RAPD marker and HPLC.

    Science.gov (United States)

    Chetri, Siva K; Sardar, Pratima Rani; Agrawal, Veena

    2014-10-01

    In vitro protocol has been established for clonal propagation of Cassia angustifolia Vahl which is an important source of anticancerous bioactive compounds, sennoside A and B. Nodal explants excised from field raised elite plant (showing optimum level of sennoside A and B) of C. angustifolia when reared on Murashige and Skoog's medium augmented with different cytokinins, viz. N(6)-benzyladenine (BA), N(6)-(2-isopentenyl) adenine (2iP) and 6-furfuryl aminopurine (Kn) differentiated multiple shoots in their axils. Of the three cytokinins, BA at 5 μM proved optimum for differentiating multiple shoots in 95 % cultures with an average of 9.14 shoots per explant within 8 weeks of culture. Nearly, 95 % of the excised in vitro shoots rooted on half strength MS medium supplemented with 10 μM indole-3-butyric acid (IBA). The phenotypically similar micropropagated plants were evaluated for their genetic fidelity employing random amplified polymorphic DNA (RAPD) markers. Eleven individuals, randomly chosen amongst a population of 120 regenerants were compared with the donor plant. A total of 36 scorable bands, ranging in size from 100 to 1,000 bp were generated amongst them by the RAPD primers. All banding profiles from micropropagated plants were monomorphic and similar to those of mother plant proving their true to the type nature. Besides, high performance liquid chromatography evaluation of the sennoside A and B content amongst leaves of the mature regenerants and the elite mother plant too revealed consistency in their content.

  15. Comparative evaluation of genetic diversity using RAPD, SSR and cytochrome P450 gene based markers with respect to calcium content in finger millet (Eleusine coracana L. Gaertn.)

    Indian Academy of Sciences (India)

    Preety Panwar; Manoj Nath; Vijay Kumar Yadav; Anil Kumar

    2010-08-01

    Genetic relationships among 52 Eleusine coracana (finger millet) genotypes collected from different districts of Uttarakhand were investigated by using randomly amplified polymorphic DNA (RAPD), simple sequence repeat (SSR) and cytochrome P450 gene based markers. A total of 18 RAPD primers, 10 SSR primers, and 10 pairs of cytochrome P450 gene based markers, respectively, revealed 49.4%, 50.2% and 58.7% polymorphism in 52 genotypes of E. coracana. Mean polymorphic information content (PIC) for each of these marker systems (0.351 for RAPD, 0.505 for SSR and 0.406 for cyt P450 gene based markers) suggested that all the marker systems were effective in determining polymorphisms. Pair-wise similarity index values ranged from 0.011 to 0.999 (RAPD), 0.010 to 0.999 (SSR) and 0.001 to 0.998 (cyt P450 gene based markers) and mean similarity index value of 0.505, 0.504 and 0.499, respectively. The dendrogram developed by RAPD, SSR and cytochrome P450 gene based primers analyses revealed that the genotypes are grouped in different clusters according to high calcium (300–450 mg/100 g), medium calcium (200–300 mg/100 g) and low calcium (100–200 mg/100 g). Mantel test employed for detection of goodness of fit established cophenetic correlation values above 0.95 for all the three marker systems. The dendrograms and principal coordinate analysis (PCA) plots derived from the binary data matrices of the three marker systems are highly concordant. High bootstrap values were obtained at major nodes of phenograms through WINBOOT software. Comparison of RAPD, SSR and cytochrome P450 gene based markers, in terms of the quality of data output, indicated that SSRs and cyt P450 gene based markers are particularly promising for the analysis of plant genome diversity. The genotypes of finger millet collected from different districts of Uttarakhand constitute a wide genetic base and clustered according to calcium contents. The identified genotypes could be used in breeding programmes and

  16. Similaridade genética de variedades crioulas de arroz, em função da morfologia, marcadores RAPD e acúmulo de proteína nos grãos Genetic similarities of rice landraces according to morphology, RAPD markers and grain protein content

    Directory of Open Access Journals (Sweden)

    Rogeria Gregio de Biase Martins Areias

    2006-01-01

    Full Text Available Em experimentos em casa de vegetação e câmara de crescimento, estudou-se o possível aparecimento de réplicas em 20 variedades crioulas de arroz do Maranhão, mantidas no Banco Ativo de Germoplasma (BAG, do Centro Nacional de Pesquisa em Arroz e Feijão (CNPAF. As vinte variedades, divididas em seis grupos, com nomes similares e números de acessos diferentes no BAG foram avaliadas quanto à similaridade morfológica e molecular. As características morfológicas foram avaliadas e utilizadas para a construção de um dendrograma de similaridade. Após a colheita determinou-se o teor de proteína bruta dos grãos. No experimento em câmara de crescimento além das vinte variedades, utilizou-se a cultivar melhorada (IAC47. O DNA da parte aérea dessas plantas foi amplificado pela técnica do RAPD (DNA Polimórfico Amplificado ao Acaso, para se avaliar a similaridade genética entre as variedades. O dendrograma utilizando as características morfológicas revelou dois grupos, sendo um formado pelas plantas de nome "Lageado" e outro com as demais variedades. A análise genética confirmou os dados de morfologia, separando o grupo "Lageado" (com mais de 70 % de similaridade das demais variedades, que formaram um grupo com dois subgrupos. Em um dos subgrupos, agruparam as variedades com maiores teores de proteína bruta nos grãos, confirmando a alta diversidade das variedades estudadas. Os resultados revelaram que algumas plantas de nome semelhante e número de acesso distinto são provavelmente a mesma variedade, e as plantas de nome "Lageado" formam um grupo à parte, cujas características, inclusive produtividade, merecem maiores estudos.Twenty rice landraces from Centro Nacional de Pesquisa em Arroz e Feijão (CNPAF, Brazil, were studied as to morphological traits, DNA markers revealed by RAPD technique and protein content. Assessed landraces were different Active Germplasm Bank (AGB accessions comprising six groups of same common names

  17. Analysis of the genetic diversity in Metopolophium dirhodum (Walker (Hemiptera, Aphididae by RAPD markers Análise da diversidade genética de Metopolophium dirhodum (Walker (Hemiptera, Aphididae por meio de marcadores RAPD

    Directory of Open Access Journals (Sweden)

    Marcelo Lopes-da-Silva

    2007-03-01

    Full Text Available The emergence of host-races within aphids may constitute an obstacle to pest management by means of plant resistance. There are examples of host-races within cereals aphids, but their occurrence in Rose Grain Aphid, Metopolophium dirhodum (Walker, 1849, has not been reported yet. In this work, RAPD markers were used to assess effects of the hosts and geographic distance on the genetic diversity of M. dirhodum lineages. Twenty-three clones were collected on oats and wheat in twelve localitites of southern Brazil. From twenty-seven primers tested, only four primers showed polymorphisms. Fourteen different genotypes were revealed by cluster analysis. Five genotypes were collected only on wheat; seven only on oats and two were collected in both hosts. Genetic and geographical distances among all clonal lineages were not correlated. Analysis of molecular variance showed that some molecular markers are not randomly distributed among clonal lineages collected on oats and on wheat. These results suggest the existence of host-races within M. dirhodum, which should be further investigated using a combination of ecological and genetic data.A emergência de raças hospedeiro-especialistas em afídeos pode constituir um obstáculo ao manejo de pragas por meio de plantas resistentes. Existem exemplos de raças hospedeiro-especialistas em afídeos de cereais, embora a ocorrência de raça hospedeiro-especialista no pulgão-verde-pálido-do-trigo Metopolophium dirhodum (Walker, 1849 (Hemiptera, Aphididae não tenha sido relatada ainda. Marcadores RAPD foram utilizados para avaliar os efeitos da distância geográfica e do hospedeiro sobre a diversidade genética de linhas clonais de M. dirhodum. Vinte e três clones foram coletados em aveia e trigo em doze localidades do sul do Brasil. De vinte e sete iniciadores usados para a análise, apenas quatro iniciadores mostraram polimorfismos. A análise de agrupamento por similaridade genética revelou haver quatorze

  18. Divergência genética em tomate estimada por marcadores RAPD em comparação com descritores multicategóricos Genetic divergence among tomato accessions using RAPD markers and its comparison with multicategoric descriptors

    Directory of Open Access Journals (Sweden)

    Leandro SA Gonçalves

    2008-09-01

    Full Text Available A estimativa da variabilidade genética existente em um banco de germoplasma é importante não só para a conservação dos recursos genéticos, mas também para aplicações no melhoramento de plantas. O presente trabalho teve como objetivo estudar a divergência genética entre 78 acessos de uma coleção de germoplasma de tomateiro, com base em 74 marcadores RAPD e correlacionar esses resultados àqueles da caracterização morfoagronômica realizada para 27 descritores. Foi utilizado o agrupamento hierárquico UPGMA para analisar os dados, observando-se a formação de 13 grupos. Esses grupos foram correlacionados a cinco descritores (hábito de crescimento, tipo de folha, cor do fruto, número de lóculos e formato do fruto. Alguns grupos apresentaram peculiaridades, a exemplo do grupo IV, que reuniu acessos com frutos no formato de pêra; o grupo VII com acessos resistentes a murcha-bacteriana e o grupo IX, que englobou acessos com folhas do tipo batata. As análises por bootstrap revelaram poucos agrupamentos consistentes. Houve correlação positiva e altamente significativa entre as matrizes geradas pelos 27 descritores qualitativos e pelos marcadores RAPD (t = 14,02. A correlação de Mantel (r = 0,39 foi altamente significativa, porém de baixa magnitude. O baixo valor verificado para esta correlação sugere que ambas as etapas de caracterização (morfoagronômica e molecular são importantes para um conhecimento mais amplo e melhor discriminação entre os acessos de tomate.The estimation of genetic variability in a germplasm bank is important not only for the conservation of the genetic resources, but also for applications in plant breeding. The genetic divergence among 78 tomato accessions was studied, based on 74 RAPD markers. Also, a correlation between the molecular profile and 27 morphological and agronomic data was performed. Cluster analysis (UPGMA, used to study the data, resulted in 13 groups that were correlated with

  19. Genetic structure of annual ryegrass (Lolium multiflorum populations estimated by RAPD Estrutura genética de populações de azevém anual (Lolium multiflorum estimada por RAPD

    Directory of Open Access Journals (Sweden)

    Eduardo Alano Vieira

    2004-01-01

    Full Text Available Annual ryegrass is a temperate climate annual foraging grass, grown mostly in the South of Brazil, especially in the state of Rio Grande do Sul. Despite its importance, little is known about its genetic diversity, both within and among the populations cultivated. This knowledge is of fundamental importance for developing breeding and conservation strategies. The aim of this study was to characterize the genetic diversity and structure of four populations of annual ryegrass. Three of the populations were located in Rio Grande do Sul and the fourth in Uruguay. RAPD markers were used to study the genetic diversity and structure of these populations. Analysis of 375 individuals sampled from the populations, using six RAPD primers, generated a total of 82 amplified bands. They included 73 polymorphic bands (89,02%. The value of the total genetic diversity index obtained, (0,71 was high, indicating the presence of wide genetic diversity in the four populations. Genetic structure analysis revealed that 98% of total diversity is intrapopulational, whereas interpopulational genetic diversity was only 2%. These results suggest that before these populations separated, they had gone through a period of gene exchange and, even after the separation event, gene frequency stayed at levels similar to the original levels, with no differential selection for these genes in the different cultivation areas.O azevém anual é a gramínea anual forrageira de clima temperado de maior utilização no sul do Brasil, particularmente no Estado do Rio Grande do Sul. Apesar de toda a importância que a espécie apresenta, pouco se conhece a respeito da diversidade genética presente entre e dentro das populações cultivadas. Este conhecimento é de fundamental importância para o estabelecimento das estratégias de melhoramento genético e de conservação destes materiais. O objetivo deste estudo foi caracterizar a diversidade genética e a estrutura genética de quatro

  20. RAPD在酵母分类鉴定与育种研究中的应用%Application of RAPD in Classification,Identification and Breeding of Yeasts

    Institute of Scientific and Technical Information of China (English)

    冯敏; 刘延琳

    2012-01-01

    随机扩增多态性DNA(RAPD)因其简便、快速、灵敏等优点在酵母的分类鉴定,种属间亲缘关系确定及育种等方面有广阔的应用前景。本文综述近年来RAPD技术在酵母相关研究中的应用,并简单讨论该技术目前的研究热点。%Random amplified polymorphic DNA(RAPD) has a promising prospect for applications in yeast classification and identification,genetic relationship confirmation among yeast species,and breeding.It is a rapid and sensitive method to analyze microbial characters.In this paper,the recent applications of RAPD in the classification,identification and breeding of yeasts are summarized and the current research hot topics on RAPD are also discussed.

  1. RAPD and SCAR markers as potential tools for detection of milk origin in dairy products: Adulterant sheep breeds in Serra da Estrela cheese production.

    Science.gov (United States)

    Cunha, Joana T; Ribeiro, Tânia I B; Rocha, João B; Nunes, João; Teixeira, José A; Domingues, Lucília

    2016-11-15

    Serra da Estrela Protected Designation of Origin (PDO) cheese is the most famous Portuguese cheese and has a high commercial value. However, the adulteration of production with cheaper/lower-quality milks from non-autochthones ovine breeds compromises the quality of the final product and undervalues the original PDO cheese. A Randomly Amplified Polymorphic DNA (RAPD) method was developed for efficient detection of adulterant breeds in milk mixtures used for fraudulent production of this cheese. Furthermore, Sequence Characterized Amplified Region (SCAR) markers were designed envisioning the detection of milk adulteration in processed dairy foods. The RAPD-SCAR technique is here described, for the first time, to be potentially useful for detection of milk origin in dairy products. In this sense, our findings will play an important role on the valorization of Serra da Estrela cheese, as well as on other high-quality dairy products prone to adulteration, contributing to the further development of the dairy industry.

  2. Genetic relationship among Camponotus rufipes Fabricius (Hymenoptera: Formicidae nests by RAPD molecular markers - doi: 10.4025/actascibiolsci.v35i1.10913

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    Alexandre Wagner Silva Hilsdorf

    2012-12-01

    Full Text Available Random amplified polymorphic DNA (RAPD markers were used to investigate the genetic relationship among nests of the carpenter ant, Camponotus rufipes, located in the same area. Five random oligodecamers were used to amplify DNA from 108 ant workers collected from six nests. A total of 47 RAPD markers were identified, which revealed low levels of genetic differentiation among nests (Fst = 0.00218 and a low average Shannon index (0.3727 among workers within nests. These results together suggest that the C. rufipes nest may be formed by a single, once-mated queen and that nests produced by queens that are genetically related tend to keep their nests in close proximity to one other.

  3. Identificação de parentais e híbridos entre Vitis vinifera e Vitis rotundifolia utilizando polimorfismo enzimático e marcador RAPD Identification of parents and hybrids among Vitis vinifera and Vitis rotundifolia using isozyme polymorphism and rapd marker

    Directory of Open Access Journals (Sweden)

    Haiko Enok Sawazaki

    1996-01-01

    Full Text Available Identificaram-se parentais e híbridos entre Vitis vinifera (videiras comuns e V. rotundifolia (muscadínias, utilizando-se o polimorfismo enzimático e marcador RAPD (Random Amplified Polymorphic DNA. Os sistemas GOT (glutamato-oxalo-acetato-transaminase, IDH (isocitrato desidrogenase e PGI (fosfoglucose isomerase diferenciaram a muscadínia, sendo observadas cinco aloenzimas para o GOT, duas para o LAP (leucina aminopeptidase e quatro para o EST (esterase. Os sistemas PGI e IDH apresentaram-se como diméricos com o fenótipo de quatro aloenzimas em duas regiões e três em uma região respectivamente. O marcador RAPD apresentou polimorfismo que permitiu a diferenciação entre todos os cultivares. Os dendrogramas UPGMA (unweighted pair-group method with aritmetic mean obtidos pelas isoenzimas e pelo marcador RAPD foram semelhantes, sendo a aproximação mais forte entre 'Itália' e 'Rubi' que se ligaram aos cultivares Patrícia e A Dona. Os cultivares Piratininga e Eugênio, também bastante próximos, foram os seguintes a se ligarem às demais viníferas. Pelo polimorfismo enzimático e marcador RAPD, a muscadínia ficou bastante isolada dos outros grupos. Pelo método RAPD, aplicado às muscadínias, ao híbrido da Carolina do Norte NC66 C203-9, a um possível híbrido e seu parental feminino, observou-se o seguinte: os híbridos foram intermediários às muscadínias e viníferas, porém o possível híbrido se assemelhou ao parental feminino, enquanto o NC66203-9 apresentou bandas provenientes das muscadínias e viníferas, comprovando sua origem híbrida.Aiming to identify parents and hybrids among Vitis vinifera (bunch grapes and Vitis rotundifolia (muscadine grapes isozyme polymorphism and RAPD marker were used. GOT (glutamate oxaloacetate transaminase, IDH (isocitrate dehydrogenase and PGI (phosphoglucoisomerase systems could differentiate the muscadine, being identified five allozymes for GOT, two allozymes for LAP (leucine

  4. Assessment of genetic diversity and relationships among Egyptian mango (Mangifera indica L.) cultivers grown in Suez Canal and Sinai region using RAPD markers.

    Science.gov (United States)

    Mansour, Hassan; Mekki, Laila E; Hussein, Mohammed A

    2014-01-01

    DNA-based RAPD (Random Amplification of Polymorphic DNA) markers have been used extensively to study genetic diversity and relationships in a number of fruit crops. In this study, 10 (7 commercial mango cultivars and 3 accessions) mango genotypes traditionally grown in Suez Canal and Sinai region of Egypt, were selected to assess genetic diversity and relatedness. Total genomic DNA was extracted and subjected to RAPD analysis using 30 arbitrary 10-mer primers. Of these, eleven primers were selected which gave 92 clear and bright fragments. A total of 72 polymorphic RAPD bands were detected out of 92 bands, generating 78% polymorphisms. The mean PIC values scores for all loci were of 0.85. This reflects a high level of discriminatory power of a marker and most of these primers produced unique band pattern for each cultivar. A dendrogram based on Nei's Genetic distance co-efficient implied a moderate degree of genetic diversity among the cultivars used for experimentation, with some differences. The hybrid which had derived from cultivar as female parent was placed together. In the cluster, the cultivars and accessions formed separate groups according to bearing habit and type of embryo and the members in each group were very closely linked. Cluster analysis clearly showed two main groups, the first consisting of indigenous to the Delta of Egypt cultivars and the second consisting of indigenous to the Suez Canal and Sinai region. From the analysis of results, it appears the majority of mango cultivars originated from a local mango genepool and were domesticated later. The results indicated the potential of RAPD markers for the identification and management of mango germplasm for breeding purposes.

  5. Study on Selected Mating of Highbrid Combination by RAPD Markers in Pig%用RAPD标记选配猪杂交组合的研究

    Institute of Scientific and Technical Information of China (English)

    赵凯; 张周平; 杨公社; 周继平; 魏雅萍

    2001-01-01

    With the help of Random Amplified PolymorphicDNA(RAPD)Markers,the genetic distances between parents,form 10 porcine breeds cross combinations were measured.And the coefficient between these results and such traits of F1 as the growing and fattening and carcass performance were also discussd.The results showed:the genetic obtained by RAPD marker were positively related to average daily gain of F1,dressing percentage,lean percentage,significantely,and were negatively related to average backfat thickness,feed efficiency,significantely.Therefore,RAPD can be used in a marker subsidiary selection on parental selcted mating of hybridization.%利用随机扩增多态DNA(RAPD)技术测定了10个猪种(含3个杂种)9个杂交组合亲本间的遗传距离,并对这些参数与F1生长肥育性状和胴体性状的相关性进行了分析。结果表明:RAPD标记遗传距离与F1平均日增重呈极显著正相关,与屠宰率和瘦肉率呈显著正相关,与平均背膘厚、饲料转化率呈显著负相关。因此RAPD作为标记辅助选择方法可用于选配杂交组合。

  6. Fronteras del corazón shamánico. Azares y dilemas Qom. En: Avá, nº 16

    OpenAIRE

    2009-01-01

    En este artículo analizo dilemas y paradojas que plantea la modernidad en el caso de los Takshek Qom o tobas del oriente de Formosa (Argentina). Para ello me centro en la experiencia del lkillakte o corazón como uno de los núcleos vitales y existenciales de la persona. Allí se alojan las distintas clases de poder shamánico y también es sede de los pensamientos y las emociones. Me interesa explorar qué le pasa en términos simbólicos al corazón shamánico frente al ...

  7. Development and significance of SCAR marker QG12-5 for Canarium album (Lour.) Raeusch by molecular cloning from improved RAPD amplification.

    Science.gov (United States)

    Cheng, J L; Yin, Z C; Mei, Z Q; Wei, C L; Chen, H C; Wu, X S; Fu, J J

    2016-08-26

    Sequence-characterized amplified region (SCAR) is a valuable molecular marker for the genetic identification of any species. This marker is mainly derived from molecular cloning of random amplified polymorphic DNA (RAPD). We have previously reported the use of an improved RAPD technique for the genetic characterization of different samples of Canarium album (Lour.) Raeusch (C. album). In this study, DNA fragments were amplified using improved RAPD amplified from different samples of C. album. The amplified DNA fragment was excised, purified from an agarose gel and cloned into a pGM-T vector; subsequently, a positive clone, called QG12-5 was identified by PCR amplification and enzymatic digestion and sequenced by Sanger di-deoxy sequencing method. This clone was revealed consisting of 510 nucleotides of C. album. The SCAR marker QG12-5 was developed using specifically designed PCR primers and optimized PCR conditions. This SCAR marker expressed seven continuous "TATG" [(TATG)n] tandem repeats, which was found to characterize C. album. Subsequently, this novel SCAR marker was deposited in GenBank with accession No. KT359568. Therefore, we successfully developed a C. album-specific SCAR marker for the identification and authentication of different C. album species in this study.

  8. [The experience of implementation of REP-u RAPD-polymerase chain reaction in epidemiologic characteristic of nosocomial isolates Pseudomonas aeruginosa].

    Science.gov (United States)

    Kuznetsova, M V; Maksimova, A V; Karpunina, T I

    2015-03-01

    The article presents comparative evaluation of diagnostic value of technique REP- u RAPD-polymerase chain reaction applied under genetic typing of clinical isolates of Pseudomonas Aeruginosa. The strains are isolated in different hospital departments of medical institutions in adult (8 medical institutions; n = 145) and children (5 medical institutions; n = 151) medical networks. The results of study demonstrated different boundary capacity of three reactions. The Simpson discrimination index made up to 0.993, 0.875 and 0.639 for RAPD-, ERIC- and BOX-polymerase chain reaction correspondingly. The RAPD-polymerase chain reaction makes it possible to detect individual characteristics of strains. Out of two alternatives the REP-polymerase chain reaction demonstrated its advantage, besides only with one primer ERIC2. The BOX-polymerase chain reaction has a least discriminating capacity under typing of isolates P. aeruginosa, detecting only species' characteristics. The clinical strains P. aeruginosa are distributed on 24 genome groups and 52 isolates had individual genotypes. The evaluation of results of genetic typing permitted to point out both similarity of tendencies in propagation of strains of P. aeruginosa among hospitalized adults and adolescents and specificity of detection in neonatal clinics. It is obvious that hospitals of different profiles, including departments of reanimation and intensive therapy represent specific ecological environment significantly different in its level of endogenous and exogenous infection.

  9. Analysis of RAPD and mitochondrial 16S rRNA gene sequences from Trichiurus lepturus and Eupleurogrammus muticus in the Yellow Sea

    Institute of Scientific and Technical Information of China (English)

    MENG Zining; ZHUANG Zhimeng; JIN Xianshi; TANG Qisheng; SU Yongquan

    2004-01-01

    Random amplified polymorphic DNA (RAPD) technique is applied to 12 individuals from each species of the hairtail fishes Trichiurus lepturus and Eupleurogrammus muticus in the Yellow Sea. The percentage of polymorphic sites, degree of genetic polymorphism and genetic distance are compared and the phylogenetic tree is constructed by Neighbor-joining method. The partial mitochondrial 16S rRNA gene is amplified by polymerase chain reaction (PCR) and the PCR products are directly sequenced after being purified. These sequences, together with the homologous sequences of another Trichiuridae species Lepidopus caudatus obtained from GenBank, are used to analyze nucleotide difference and to construct a UPGMA phylogenetic tree by means of biological informatics. Analysis shows: (1) the RAPD technique is a highly sensitive method for investigating genetic diversity in T. lepturus, and E. muticus. T. lepturus exhibits a lower polymorphism and genetic diversity than E. muticus; (2) according to the analysis of the partial mitochondrial 16S rRNA gene sequences, a very low intraspecific variation and considerably high divergence among species were found, which reveals a dual nature of conservatism and variability in mitochondrial 16S rRNA gene; (3) five primers generate the species-specific RAPD sites and these sites can be served as the molecular markers for species identification and (4) it can be proved at DNA variation level that T. lepturus and E. muticus are of two species respectively pertaining to different genera, which supports the Nelson taxonomic conclusion.

  10. The RAPD analysis of Juglans nigra cultivars introduced in Ningxia%宁夏引种黑核桃品种RAPD分析研究

    Institute of Scientific and Technical Information of China (English)

    李苗; 黄学琴; 吴全忠; 张丽; 陈虞超; 甘晓燕; 石磊; 宋玉霞

    2011-01-01

    33 pieces of Juglans nigra and 1 Juglans were used as experimental materials,the developed CTAB method was used to extract high quality DNA,and RAPD-PCR by 24 primers was employed to analyze the genetics relationship between the experimental materials.The result showed that 18 RAPD primers could be used in appraise Juglans nigra genetics resource and clustering the experimental materials correctly.Genetics relationship date showed that 33 experimental materials of Juglans nigra was farer than Juglans,nevertheless closer genetics relationship among Juglans nigra experimental materials.%以宁夏引种种植的33份黑核桃树种和1份本地核桃树种为试验材料,采用改进的CTAB法提取高质量模板DNA,通过24条RAPD引物利用RAPD-PCR技术系统分析材料间的遗传关系.结果表明:试验所筛选的18条RAPD引物可以用于黑核桃树种遗传资源鉴定,较为准确地将种间材料聚类;33份黑核桃材料与作为对照参试的普通核桃亲缘关系较远,参试黑核桃材料间则表现出较近的亲缘关系.

  11. Standardization and validation of simple PCR, duplex PCR and RAPD in comparison to blood smear examination for diagnosing bovine tropical theileriosis.

    Science.gov (United States)

    Sudan, Vikrant; Shanker, Daya; Jaiswal, Amit; Singh, Amit; Pandey, Vijay

    2017-03-01

    Bovine Tropical Theileriosis (BTT) is an important vector-borne protozoan disease that imposing serious constraints on the health and productivity of domestic cattle. It is matter of common fact that following recovery from primary infection, cattle become persistent carriers and act as reservoirs of infection thereby, playing a critical role in disease epidemiology. The present study describes the comparative diagnostic efficiency of simplex PCR, duplex PCR and RAPD assays for detection of Theileria annulata in cattle. An optimized simple PCR and duplex PCR assay were established using TAMS F/R as primer sets encoding for 721 bp amplicon alongside a RAPD with arbitrary primer coding for 963 bp product of T. annulata. The simple PCR and duplex PCR detected pathogen with almost same level of sensitivity, irrespective of whether its DNA was amplified in isolation or together with DNA of another pathogen without nonspecific amplifications. RAPD failed to give comparable results and suffered from limitations of sensitivity as well as specificity. The developed assays may be seen as a good tool for epidemiological studies aiming at assessing the burden of chronic infections and improving control of the associated diseases in endemic regions.

  12. Localization of genes for lateral branch and female sex expression and construction of a molecular linkage map in cucumber (Cucumis sativus L. ) with RAPD markers

    Institute of Scientific and Technical Information of China (English)

    LI Xiaozun; PAN Junsong; WANG Gang; TIAN Libo; SI Longting; WU Aizhong; CAI Run

    2005-01-01

    A cucumber ( Cucumis sativus L. ) molecular linkage map, including 79 random-amplified polymorphic DNAs (RAPD)and two genes , lb for lateral branch and f for female sex expression, is constructed from a cross between a line, S52, with weak lateral growing ability and staminate from Dabieshan Mountains area in China and another line, S06, with strong lateral growing ability and gynoecious from Europe. The map contains nine linkage groups and spans 1110.0 cM with an average distance of 13.7 cM between loci. The lb locus is located in a longer linkage group LG-2 and flanked by two markers, OP-Q5-1 and OP-M-2-2, at 9.3 cM and 15.9 cM, respectively. In the meantime, the RAPD loci, OP-Q5-2 and BC151, in a short linkage group were found to flank f at 13.7 cM and 13.4 cM,respectively. The construction of RAPD map has paved a way for further study of the genes for lateral branch, female sex expression and other agronomic traits in cucumber.

  13. RAPD-PCR typing of Yersinia enterocolitica (Enterobacteriaceae O:3 serotype strains isolated from pigs and humans

    Directory of Open Access Journals (Sweden)

    Tereza Cristina A. Leal

    1999-09-01

    Full Text Available Sixteen strains of Yersinia enterocolitica serotype O:3, isolated from apparently healthy pigs collected in Rio de Janeiro, and four human strains of serotypes O:4, O:5, O:6 and O:13 were analyzed by RAPD-PCR. The strains were grouped into five genotypic profiles according to the amplification patterns obtained with three random primers. Fifteen of the 16 pig strains had identical amplification patterns, which was named genotypic profile 1. The one different profile was named genotypic profile 2. Genotypic profile 1 was also exhibited by the O:6 human serotype strain. The O:4 and O:13 human serotype strains showed similar amplification profiles with two primers. However, the third primer induced a distinct profile in each strain. Therefore, these two strains were placed into genotypic profile 3 and 4, respectively. Each primer produced a completely different amplification profile in the O:5 human serotype strain; therefore, it was named genotypic profile 5. The presence or absence of plasmids in the strains studied did not affect the amplification results. These results show that genetic variations can exist within a serotype, and strains of different serotypes can exhibit the same amplification profile when compared using other primers.Foram utilizados três "primers" aleatórios para caracterizar pela técnica RAPD-PCR 16 cepas de Yersinia enterocolitica do sorotipo O:3, isoladas de suínos sadios do Rio de Janeiro. Pelos resultados dos padrões de amplificação, as 16 cepas dos suínos e as 4 cepas humanas usadas como referência (sorotipos O:4, O:5, O:6 e O:13 foram agrupadas em 5 perfis genotípicos. Quinze cepas de suínos apresentaram um padrão de amplificação idêntico (perfil genotípico 1 e somente uma apresentou um perfil de amplificação diferente (perfil genotípico 2. O mesmo padrão de amplificação do perfil genotípico 1 foi também observado em uma cepa humana do sorotipo O:6. As cepas humanas dos sorotipos O:4 e O:13

  14. An efficient and rapid DNA minipreparation procedure suitable for PCR/SSR and RAPD analyses in tropical forest tree species

    Directory of Open Access Journals (Sweden)

    Ana Lilia Alzate-Marin

    2009-10-01

    Full Text Available An efficient and rapid DNA minipreparation modified method for frozen samples was developed for five tropical tree species: Copaifera langsdorffii, Hymenaea courbaril, Eugenia uniflora, Tabebuia roseo alba and Cariniana estrellensis. This procedure that dispenses the use of liquid nitrogen, phenol and the addition of proteinase K, is an adaptation of the CTAB-based DNA extraction method. The modifications included the use of PVP to eliminate the polyphenols, only one chloroform-isoamyl alcohol step and the addition of RNase immediately after extraction with chloroform. The yields of the DNA samples ranged from 25.7 to 42.1 µg from 100 mg leaf tissue. The DNA samples extracted by this method were successfully used for PCR (SSR and RAPD analyses in these five and other twelve tropical tree species.Este trabalho teve como objetivo otimizar um protocolo econômico, rápido e eficaz de minipreparação de DNA genômico, para as espécies florestais Copaifera langsdorffii (Óleo de Copaíba, Hymenaea courbaril (Jatobá, Eugenia uniflora (Pitanga, Tabebuia roseo alba (Ipê Branco e Cariniana estrellensis (Jequitibá Branco. Este método é uma adaptação da técnica de extração CTAB de Doyle e Doyle (1990, o qual consiste principalmente na adição de PVP para eliminar polifenoles, somente uma etapa de extração com clorofórmio-álcool isoamílico e a adição da RNase A imediatamente após a extração com clorofórmio. O método também dispensa o uso de nitrogênio líquido, o uso do fenol e a adição de proteinase K. Os DNAs das espécies florestais extraídos apresentaram alto rendimento e boa qualidade, com rendimento de 25.7 a 42.1 µg de DNA a partir de 100 mg de tecido foliar congelado. Com este protocolo, em apenas 1 dia de trabalho, uma pessoa pode completar o isolamento do DNA de aproximadamente 50 amostras de folhas (dependendo da capacidade da centrífuga. O DNA obtido pode ser usado para métodos de análise baseados em PCR (SSR e

  15. Extraction of DNA and Optimization of RAPD-PCR Protocol in 11 Species of Cymbidium%11种兰属植物DNA的提取及RAPD-PCR实验体系的建立与优化

    Institute of Scientific and Technical Information of China (English)

    王国鼎; 文晓鹏; 季祥彪; 乔光; 胡鹏

    2007-01-01

    采用改进的CTAB-DNA提取方法,从11种兰属植物的嫩叶中提取总DNA.所得的DNA样品的A260/A280值在1.7~1.9之间,琼脂糖凝胶上主带清晰,较少降解,样品纯度高,DNA量大.另外,对影响RAPD-PCR的Mg2+、dNTPs、Taq酶、引物浓度等因素进行了优化.确定优化的反应体系为:75ng模板DNA,1×Buffer,2.5mmol/L Mg2+,0.15mmol/L dNTPs,0.75U Taq酶,引物浓度0.4μmol/L,反应总体积为25μl.该体系在20个供试兰属实验材料中获得较好的扩增结果.

  16. RANDOM AMPLIFIED POLYMORPHIC DNA (RAPD) REVEALED THE HETEROKARYON OF AGARICUS ARVENSIS PRODUCED BY MATING REACTIONS%随机扩增多态DNA(RAPD)在野蘑菇杂交育种中的应用

    Institute of Scientific and Technical Information of China (English)

    李荣春; 张丽梅

    2001-01-01

    野蘑菇(Agaricus arvensis Schaeff ex.Fr.)是蘑菇属(Agaricus)又一种近年来广泛栽培的食用菌.由于它的菌丝细胞具有多核,无锁状联合,以及人们对它的繁殖模式和生活史认识的不足,给杂交育种工作造成了较多困难.运用随机扩增多态DNA遗传标记技术,结合拮抗试验和核相分析,对自育的单孢菌株之间的杂交试验进行分析研究.结果表明:两个相互亲合的同核体菌株被配对培养时,交配反应出现,并形成异核体的后代.可能野蘑菇具有双重的交配繁殖系统--同宗配合和异宗配合.在食用菌杂交育种研究中,机扩增多态DNA(RAPD)是一种非常有效和方便的检验杂合子的方法.

  17. Identification of a locus characteristic of male individuals of buffalo grass [Buchloe dactyloides (Nutt.) Engelm.] by using an RAPD marker.

    Science.gov (United States)

    Li, Y X; Wang, X G; Yang, C H; Cong, L L; Wu, F F; Xue, J G; Han, Y H

    2013-09-27

    Buffalo grass [Buchloe dactyloides (Nutt.) Engelm.] plants can be either male, female, or hermaphrodite (monoecious). As there is no morphological difference in the early vegetative growth of these three classes of plants, it is worthwhile to use molecular biological methods to attempt to identify the sex of a plant at this early growth period. In this study, we identified 23 plants that had a stable sex for over at least 3 years. Of these, 9 were male plants, 10 were female plants, and 4 were hermaphrodites. Screening of 300 RAPD primers identified a primer, namely S211 (5'-ttccccgcga-3'), which is capable of identifying male plants. The specific fragment was cloned, sequenced, and submitted to the GenBank database (accession No. JN982469). When used to identify the sex of 188 plants during their first growing season, the S211 primer correctly identified 85.8% of all male plants. Our results showed that the S211 primer can identify the male, and in doing so, it facilitates buffalo grass breeding work.

  18. Molecular Markers for Identification of Stellantchasmus falcatus and a Phylogenic Study using the HAT-RAPD Method

    Science.gov (United States)

    Wongsawad, Pheravut

    2010-01-01

    Stellantchasmus falcatus is a minute intestinal fluke in the family Heterophyidae. Metacercariae, the infective stage, were reported in a marine fish, mullet Liza subviridis, and a fresh water fish, Dermogenus pusillus, in Thailand. Adults were found in chicks, rats, cats, and humans. Morphological studies were done for comparing Stellantchasmus sp. worms found in 2 different fish hosts; their shapes and organ arrangements were very similar except for the prepharynx length. Therefore, the present study aimed to compare their DNA fingerprints using the HAT-RAPD method for both types of Stellantchasmus and several other related species. Ten arbitrarily selected primers (OPA-04, OPA-09, OPN-02, OPN-03, OPN-09, OPN-12, OPP-11, OPR-15, OPX-13, and OPAD-01) were used. It was found that OPA-09, OPN-03, and OPAD-01 were able to generate S. falcatus specific fragments in mullets which consisted of 200, 760, and 280 bp, respectively. In addition, the results of morphologic, DNA fingerprinting, and phylogenetic analyses strongly suggest that the fresh water and marine specimens of Stellantchamus may be different species. PMID:21234232

  19. Analysis of genetic diversity of cultivars and clones of white clover (Trifolium repens L. with the use of RAPD markers

    Directory of Open Access Journals (Sweden)

    Agnieszka Tomkowiak

    2012-12-01

    Full Text Available The research focused on four white clover cultivars (which are synthetic clones and one cultivar generatively propagated through the selection-propagation school. The overall objective of this study was to analyse the genetic diversity of cultivars and clones of white clover and to determine the share of individual clones in the creation of cultivars based on genetic similarity specified using RAPD molecular markers. For each similarity coefficient the similarity matrix was created. Then the dendrograms were built. For the results comparison for each ratio Mantel test and Spearman correlation coefficient were used. On the basis of the analysis it was stated that selected primers generated polymorphisms, which allowed to select components for the crosses to test the ability of combinations. The dendrograms constructed with the use of the coefficients of Nei and Ochiai create similarity groups which include cultivars with their clones so they the most precisely group the forms in terms of the origin. Coefficients of Simple Matching, Hamann, Rogers and Tanimoto create groups, which often contain cultivars and clones that do not belong to a given cultivar, so they are not so useful in selecting components for the crosses.

  20. Sex Identification of Pistachio by Using RAPD Analysis%阿月浑子性别鉴定的RAPD分析

    Institute of Scientific and Technical Information of China (English)

    谭冬梅; 罗淑萍; 李疆; 韩海涛

    2003-01-01

    应用随机扩增多态性DNA(RAPD)技术对新疆阿月浑子(Pistacia vera L.)地方品种雌、雄株进行性别鉴定的研究.通过叶片总DNA抽提、RAPD标记分析及反复的试验筛选,找到了一个适用于新疆阿月浑子的RAPD反应体系和循环参数.采用OPO08引物对雌、雄株基因组DNA扩增出性别之间差异性的核苷酸片段,证实雄性植株DNA的扩增产物有一条大约700bp的特异性条带,而雌性植株则无此特异性条带.即找到一条与阿月浑子性别相关的基因标记,表明RAPD技术可应用于新疆阿月浑子地方品种的性别鉴定.此研究是对我国雌雄异株果树阿月浑子在分子水平进行早期性别鉴定的一个尝试.

  1. MOLECULAR CONFIRMATION OF SEX IN REGENERATED PLANTLETS OF SPINE GOURD (Momordica dioica Roxb. Ex. WILD BY USING RAPD MARKERS

    Directory of Open Access Journals (Sweden)

    S. Raju

    2015-10-01

    Full Text Available Plant tissue culture techniques offer a great opportunity to overcome the limitations associated with the large scale cultivation of spine gourd. Present study was carried out to formulate the best possible media for large scale production of spine gourd and result of the study revealed that highest percentage (85% of embryogenic callus was obtained from MS medium supplemented with 2.0 mg/L each of 2, 4- Dichlorophenoxyacetic acid (2, 4-D and 6- Benzylamino Purine (BAP in leaf explants of spine gourd. Maximum number of shoots (12.15 ± 1.51 shoots were observed on MS medium augmented with BAP (4.0 mg/L in combination with L-glutamine (2.0 mg/L from leaf derived embryogenic callus of spine gourd. Identification of sex by using morphological characters in the newly regenerated plantlets of spine gourd at fourth leaf stage is another problem for large scale propagation of female plants. PCR based molecular marker OPA-15, a Random Amplified Polymorphic DNA (RAPD primer can be used as a differential marker to identify female plants form male plants at pre-flowering stage in newly regenerated plantlets (in vitro and as well as in field plants (in vivo of spine gourd. A unique amplification band (700 bp in size appeared only in female samples, but not in male samples of spine gourd.

  2. ASSESSMENTS OF GENETIC DIVERSITY IN COUNTRY BEAN (Lablab purpureus L. USING RAPD MARKER AGAINST PHOTO-INSENSITIVITY

    Directory of Open Access Journals (Sweden)

    BISWAS MD. SANAULLAH

    2012-12-01

    Full Text Available RAPD marker was used to evaluate genetic relationships among 11 genotypes of country bean, including first three genotypes were photo-insensitive and the rests were sensitive. The genotypes were grouped into two major clusters where photo-insensitive genotypes remain in cluster I and sensitive genotypes remain in cluster II. A total of 26 bands were detected, of which 57.69% were polymorphic and the remaining were monomorphic across all genotypes. A highest level of genetic distance was observed between CB04 and CB06 while the lowest level of genetic distance showed between CB01 and CB03. The highest similarity index between the genotypes CB01 and CB03 indicated less divergence between them. Low similarity indices were observed between CB04 and CB06, which indicated more divergence. Crossing between the genotypes with low similarity coefficient will manifest high heterosis. The identified genetically distinct cultivars could be potentially important source of germplasm for further improvement of country bean.

  3. Genetic variability in the natural populations of Lasioderma serricorne (F.) (Coleoptera: Anobiidae), detected by RAPD markers and by esterase isozymes.

    Science.gov (United States)

    Coelho-Bortolo, T; Mangolin, C A; Lapenta, A S

    2016-02-01

    Lasioderma serricorne (F.) is a small cosmopolitan beetle regarded as a destructive pest of several stored products such as grains, flour, spices, dried fruit and tobacco. Chemical insecticides are one of the measures used against the pest. However, intensive insecticide use has resulted in the appearance of resistant insect populations. Therefore, for the elaboration of more effective control programs, it is necessary to know the biological aspects of L. serricorne. Among these aspects, the genetic variability knowledge is very important and may help in the development of new control methods. The objective of this study was to evaluate the genetic variability of 11 natural populations of L. serricorne collected respectively in three and four towns in the states of Paraná and São Paulo, Brazil, using 20 primers random amplified polymorphic DNA (RAPD) and polymorphisms of esterases. These primers produced 352 polymorphic bands. Electrophoretic analysis of esterases allowed the identification of four polymorphic loci (Est-2, Est-4, Est-5 and Est-6) and 18 alleles. Results show that populations are genetically differentiated and there is a high level of genetic variability within populations. The high degree of genetic differentiation is not directly correlated to geographical distance. Thus, our data indicate that movement of infested commodities may contribute to the dissemination of L. serricorne, facilitating gene flow.

  4. The assessment of genetic variability and taxonomic affi nity of local pummelo accessions from Yogyakarta, Indonesia based on RAPD

    Directory of Open Access Journals (Sweden)

    Ratna Susandarini

    2016-02-01

    Full Text Available Pummelo (Citrus maxima is one of three biotypes considered as true species within the genus Citrus.A major issue of pummelo taxonomy in Indonesia is the high number of cultivars showing variability inphenotypic characters but of unknown genetic diversity. In this study, the assessment of genetic variabilityand taxonomic affi nity of local accessions of C. maxima from Yogyakarta was examined based on RAPDfi ngerprinting. The availability of universal primers and technical simplicity makes RAPD as a molecular toolof choice for the assessment of genetic variability at various taxonomic levels. In this study, 13 accessions of C.maxima collected from Yogyakarta were observed for their genetic variability. An additional three registeredcultivars were included for comparative purpose. Two decamer primers used for the amplifi cation of DNAproduced 222 bands with 174 of them were polymorphic. The data was subjected to cluster analysis to observethe grouping of accessions and taxonomic affi nity. Results indicated high genetic variability among accessions.The dendrogram constructed using UPGMA method based on simple matching coeffi cient showed twomain clusters were which was in line to morphological characters. The grouping of accessions showed cleardifferentiation between accessions bearing white and those with reddish fruit fl esh, and thus demonstratestaxonomic value of this study in recognizing important agronomic character for this tropical fruit crop.

  5. Population Genetics of the Endemic Hawaiian Species Chrysodracon hawaiiensis and Chrysodracon auwahiensis (Asparagaceae: Insights from RAPD and ISSR Variation

    Directory of Open Access Journals (Sweden)

    Pei-Luen Lu

    2016-08-01

    Full Text Available The genus Chrysodracon has six endemic species in the Hawaii Islands. Chrysodracon hawaiiensis is endemic to Hawaii Island and was described as a distinct species in 1980. It was listed as an endangered species on the International Union for the Conservation of Nature and Natural Resources (IUCN Red List in 1997. This woody plant species was, at one time, common in exposed dry forests, but it became very rare due to grazing pressure and human development. The tree species Chrysodracon auwahiensis (C. auwahiensis, endemic to Maui and Molokai, still has large adult populations in dry lands of the islands, but unfortunately no regeneration from seed has been reported in those areas for many years. The two endemic species were examined using the molecular technique of random amplified polymorphic DNA (RAPD and inter simple sequence repeats (ISSR to determine the genetic structure of the populations and the amount of variation. Both species possess similar genetic structure. Larger and smaller populations of both species contain similar levels of genetic diversity as determined by the number of polymorphic loci, estimated heterozygosity, and Shannon’s index of genetic diversity. Although population diversity of Chrysodracon hawaiiensis (C. hawaiiensis is thought to have remained near pre-disturbance levels, population size continues to decline as recruitment is either absent or does not keep pace with senescence of mature plants. Conservation recommendations for both species are suggested.

  6. [RAPD analysis of the intraspecific and interspecific variation and phylogenetic relationships of Aegilops L. species with the U genome].

    Science.gov (United States)

    Goriunova, S V; Chikida, N N; Kochieva, E Z

    2010-07-01

    RAPD analysis was used to study the genetic variation and phylogenetic relationships of polyploid Aegilops species with the U genome. In total, 115 DNA samples of eight polyploid species containing the U genome and the diploid species Ae. umbellulata (U) were examined. Substantial interspecific polymorphism was observed for the majority of the polyploid species with the U genome (interspecific differences, 0.01-0,2; proportion of polymorphic loci, 56.6-88.2%). Aegilops triuncialis was identified as the only alloploid species with low interspecific polymorphism (interspecific differences, 0-0.01, P = 50%) in the U-genome group. The U-genome Aegilops species proved to be separated from other species of the genus. The phylogenetic relationships were established for the U-genome species. The greatest separation within the U-genome group was observed for the US-genome species Ae. kotschyi and Ae. variabilis. The tetraploid species Ae. triaristata and Ae. columnaris, which had the UX genome, and the hexaploid species Ae. recta (UXN) were found to be related to each other and separate from the UM-genome species. A similarity was observed between the U M-genome species Ae. ovata and Ae. biuncialis, which had the UM genome, and the ancestral diploid U-genome species Ae. umbellulata. The UC-genome species Ae. triuncialis was rather separate and slightly similar to the UX-genome species.

  7. Development of a novel and efficient strategy for practical identification of Pyrus spp (Rosaceae) cultivars using RAPD fingerprints.

    Science.gov (United States)

    Lin, J; Wang, X C; Chang, Y H; Fang, J G

    2011-05-24

    Accurate and reliable cultivar identification of crop species is essential to guarantee plant material identity for purposes of registration, cultivar protection and production. To facilitate identification of plant cultivars, we developed a novel strategy for efficient recording of DNA molecular fingerprints in genotyped plant individuals. These fingerprints can be used as efficient referential information for quick plant identification. We made a random amplified polymorphic DNA (RAPD) marker analysis of 68 pear cultivars. All pear genotypes could be distinguished by a combination of eight 11-mer primers. The efficiency of the method was further verified by correct identification of four cultivars randomly chosen from the initial 68. The advantages of this identification include use of fewer primers and ease of cultivar separation by the corresponding primers marked on the cultivar identification diagram. The cultivar identification diagram can efficiently serve for pear cultivar identification by readily providing the information needed to separate cultivars. To the best of our knowledge, this is the most efficient strategy for identification of plant varieties using DNA markers; it could be employed for the development of the pear industry and for the utilization of DNA markers to identify other plant species.

  8. Effect of in vitro culture conditions on somaclonal variation in cowpea (Vigna unguiculata Walp.) using RAPD markers.

    Science.gov (United States)

    Sivakumar, P; Rajesh, S; Gnanam, R; Manickam, A

    2011-03-01

    We report a high frequency regeneration protocol in cowpea (Vigna unguiculata Walp. var. C 152) via somatic embryogenesis from 10-d-old primary leaf explants. A study was conducted to examine the effect of somaclonal variations in in vitro derived cowpea plants under field conditions. The regenerated plantlets were successfully transferred to field after hardening in vitro and grown for collecting R0, R1 and R2 seeds. The seeds of R1 and R2 generations were subsequently, grown under field conditions and their various biometrical traits were compared and evaluated with non-tissue cultured cowpea plants as check. There was no detectable somaclonal variation induced in R0-R2 in any of the biometrical traits. The results indicate that the inclusion of different plant growth promoters at specified concentrations and duration in our earlier tissue culture work did not induce any detectable mutation. The RAPD analysis also shows that there is no genetic variation among R2 cowpea plants. The somatic embryogenesis protocol we report could thus be safely applied for high frequency true-to-type regeneration and transformations protocols without any somaclonal variation.

  9. Caracterización de la diversidad genética en naranja y comparación del polimorfismo de microsatélites amplificados al azar (rams) usando electroforesis de poliacrilamida y agarosa

    OpenAIRE

    Muñoz Flores Jaime Eduardo; Vásquez Amariles Herney Darío; Caicedo Arana Alvaro; Jaramillo Vásquez Juan; Arcos Alba Lucía; Muñoz Rivera Oscar Julian; Chagüeza Villarreal Yamilet; Morillo Coronado Ana Cruz

    2010-01-01

    Se compararon las eficiencias de tres métodos de electroforesis en agarosa y poliacrilamida, usando la cámara pequeña de DNA Sequencing System y cámara grande OWL Sequi-Gen Sequencing Cell, en la detección del polimorfismo en 21 accesiones de naranja (Citrus sinensis) con empleo del cebador CGA. El gel de poliacrilamida dio mejor resolución de los productos amplificados vía PCR producidos por RAMs. Este permitió una mejor detección de bandas de ADN polimórficas, lo que facilitó la identificac...

  10. Organização do espaço urbano e expansão do calazar The organization of urban areas and expansion of kala-azar

    Directory of Open Access Journals (Sweden)

    Eduarda Ângela Pessoa Cesse

    2001-08-01

    Full Text Available OBJETIVOS: verificar determinados processos que estão relacionados com a ocupação do espaço urbano e que contribuem para a ocorrência e expansão do calazar em um município de médio porte com acentuado fluxo migratório e em expansão econômica. MÉTODOS: trata-se de um estudo epidemiológico, de corte transversal, no qual se realizou a investigação domiciliar dos casos registrados em 8 bairros e respectivos setores censitários, no município de Petrolina, Pernambuco, no período de 1992 a 1997. Está subsidiado por elementos da pesquisa laboratorial, considerando o transmissor e o reservatório. Utilizamos o modelo explicativo da determinação social do processo saúde-doença a esta situação endêmica-epidêmica. RESULTADOS: observa-se uma concentração de casos de calazar na periferia da zona urbana do município de Petrolina, em áreas de invasão e expansão, onde o saneamento básico é precário, há a presença de animais e do vetor no peridomicílio e a população apresenta um baixo grau de instrução. O sexo masculino e a faixa etária de 0-4 anos são os mais acometidos. CONCLUSÕES: os achados sugerem o estabelecimento de um novo padrão epidemiológico para o calazar em Petrolina, onde a ocorrência dessa endemia se dá em um espaço altamente modificado pela população. Tal situação caracteriza um processo de ruralização das áreas periurbanas endêmicas nas grandes cidades.OBJECTIVES: the aim of this study was to verify certain processes that are related to the occupation of urban areas and which contribute to the occurrence and expansion of kala-azar in a medium-sized town undergoing economic growth with a high influx of migrants. METHODS: the study is an epidemiological cross-section, in which house-to-house investigation was conducted concerning cases registered in 8 districts and their respective census areas, all in the municipality of Petrolina, Pernambuco state, Brazil, from 1992 to 1997. The study was

  11. Genetic characterization of the Moxotó goat breed using RAPD markers Caracterização genética da raça caprina Moxotó por marcadores RAPD

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    Rejane Rodrigues de Oliveira

    2005-03-01

    Full Text Available The objective of this study was to verify the genetic diversity between and within seven populations of Moxotó goat (n = 264 from the States of Pernambuco, Paraíba and Rio Grande do Norte, using RAPD (Random Amplified Polymorphic DNA. Moxotó, as well as other naturalized breeds, suffers genetic losses due to the indiscriminate miscegenation with breeds raised in the Northeast Region of Brazil. The genetic characterization of these genetic resources is essential to conservation and breeding programs. DNA was extracted from lymphocytes using a non-organic protocol. The 16 primers used were selected from 120 decamer oligonucleotide primers and generated 56 polymorphic bands. The analysis of molecular variance (AMOVA showed that the greater part of total genetic variability (71.55% was due to differences between individuals within populations, while 21.21% was among populations. The analysis of variance among the pairs of populations demonstrated that the populations located in Floresta, PE x Angicos, RN presented a smaller value of intrapopulational differentiation (8.9%, indicating low genetic variability among them. Nei's genetic distances varied between 0.0546 and 0.1868 in the populations. The dendrogram generated showed that the Canindé breed, used as outgroup, clustered with the populations of Moxotó, indicating a possible common origin of the naturalized goat breeds.O objetivo deste estudo foi verificar a diversidade genética entre e dentro de sete populações de cabras Moxotó (n = 264 dos Estados de Pernambuco, Paraíba e Rio Grande do Norte, usando a técnica de RAPD (Poliformismo de DNA Amplificados ao Acaso. A raça Moxotó, assim como outras naturalizadas, sofre grande erosão genética devido à miscigenação indiscriminada com outras raças existentes na Região Nordeste. A caracterização genética desta raça é essencial para programas de conservação e melhoramento. Os marcadores moleculares são uma ferramenta útil na

  12. Análisis de variabilidad genética en Moniliophthora roreri con AP-PCR y RAPD en Antioquia, Colombia Analysis of genetic variability in Moniliophthora roreri with AP-PCR and RAPD in Antioquia, Colombia

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    Afanador Kafuri Lucía

    2007-12-01

    Full Text Available Moniliophthora roreri es el agente causante de la moniliasis del cacao, la enfermedad más severa en las plantaciones de cacao en el departamento de Antioquia, Colombia. Los marcadores moleculares RAPD (Random Amplyfied Polymorphism of DNA y AP-PCR (Arbitraly Primed Polymerase Chain Reaction fueron usados para estudiar la variabilidad genética de 170 aislamientos de M. roreri colectados en doce municipios de Antioquia. El análisis dividió la población en seis grupos, el grupo G1 fue el más grande y contenía el 95% de los aislamientos con una alta similitud genética (coeficiente de similitud de 0,7 a 1, mientras los otros cinco grupos contenían solo aislamientos de Apartadó y Dabeiba con una similitud genética moderadamente baja (coeficiente de similitud entre 0,45 a 0,55. El análisis de componentes principales mostró una alta similitud genética entre la población excepto entre los aislamientos de Apartadó y Dabeiba, que registraron los más altos niveles de variabilidad genética con valores altos del índice de Shannon y el porcentaje de loci polimórficos, mientras los otros aislamientos registraron una baja variabilidad genética. Los valores de diversidad y diferenciación genética en la población muestran una introducción reciente de M. roreri en las plantaciones de cacao de Antioquia, y una reproducción predominantemente clonal en la población. De acuerdo con Amova, la mayoría de la variación genética se encontró dentro de los municipios (75,68% con solo un 5,94% presente entre las subregiones.Moniliophthora roreri is the causal agent of moniliasis in cocoa, the most severe disease affecting cocoa plantations in the Antioquia department in Colombia. RAPD (random amplified polymorphism of DNA and AP-PCR (arbitrarily-primed polymerase chain reaction molecular markers were used for studying the genetic variability of 170 M. roreri isolates collected from twelve municipalities in Antioquia. Cluster analysis divided the

  13. Diversidade genética de Enterolobium contortisiliquum (Vell. Morong. no Baixo Rio São Francisco, por meio de marcadores RAPD Genetic diversity of Enterolobium contortisiliquum (Vell. Morong. in the low San Francisco river by RAPD markers

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    Georgea da Cruz Santana

    2008-06-01

    Full Text Available Enterolobium contortisiliquum Vell. Morong (Leguminosae-Mimosoideae é uma espécie muito utilizada em programas de recuperação de matas ciliares no Baixo Rio São Francisco, devido ao seu rápido crescimento inicial. Assim, o objetivo deste trabalho foi avaliar, por meio de marcadores moleculares RAPD, a diversidade genética de oito indivíduos de uma população remanescente dessa espécie, visando contribuir para a definição de estratégias de coleta de sementes. Os indivíduos estão situados em uma área de 100 ha de mata ciliar do Baixo Rio São Francisco. Para a extração do DNA, pelo método CTAB 2%, foram utilizadas folhas tenras dos indivíduos. Testaram-se 20 oligonucleotídios de 10 bases de seqüência arbitrária, cujos produtos foram separados em gel de agarose 0,8%, submetidos à eletroforese horizontal, corados com brometo-de-etídio e visualizados em luz ultravioleta. A similaridade genética entre os indivíduos foi calculada pelo Coeficiente de Similaridade de Jaccard e a construção do dendrograma, realizada utilizando-se o método UPGMA. O valor médio de diversidade genética entre as matrizes foi de 49%, variando de 33 a 85%. Os indivíduos 6 e 7 apresentaram relativa proximidade genética (67%, não sendo indicado o plantio de suas mudas ou semeadura direta para recuperação de área ciliar em locais muito próximos. A partir dos resultados observados, podem-se desenvolver estratégias para a coleta de sementes e produção de mudas, auxiliando, assim, programas de restauração ambiental.Enterolobium contortisiliquum Vell. Morong (Leguminosae-Mimosoideae is very much used in riparian forest restoration programs in the Low San Francisco River because of its fast initial growth. The objective of this work was to evaluate by RAPD molecular markers the genetic diversity of eight individuals of a remaining population of this species, in order to contribute for the definition of strategies for seed production. The

  14. 红曲霉DNA提取及其RAPD-PCR反应体系的建立%Extraction of DNA from Monascus and determination of the optimal amplification condition for RAPD-PCR

    Institute of Scientific and Technical Information of China (English)

    袁勇芳; 许杨; 李思光; 汤祖辉

    2002-01-01

    采用改进的氯化苄法对红曲霉DNA进行提取纯化,探讨了提取液中EDTA的浓度以及其他因素对提取结果的影响.同时,以该法提取的DNA为模板,采用正交实验优化RAPD分析最佳反应条件.结果表明,当提取液中EDTA浓度为125mmol/l时,所得的红曲霉DNA的质量和数量均较理想,每克红曲霉菌丝体(湿重)能提取到50μg的DNA,分子量约为25kb,以此DNA为模板进行PCR扩增,其最佳反应体系为:Mg2+2.0mmol/l,dNTPs 0.15mmol/l,Taq 0.05U/μl,模板DNA 1.2ng/μl,随机引物 0.36μmol/l,Tris-HCl 10mmol/l pH9.0,KCl 50mmol/l,Nonidet P40 0.1%.另外,通过对比RNA酶消化前后的模板扩增结果证明了本实验酶的消化的必要性,对比了不同预变性时间处理模板对RAPD-PCR扩增的影响,发现不经过预变性的模板扩增结果最理想.

  15. Utilização de marcadores moleculares RAPD e EST's SSR para estudo da variabilidade genética em cana-de-açúcar¹ Use of molecular markers RAPD, and ESTs SSR to study genetic variability in sugarcane

    Directory of Open Access Journals (Sweden)

    João Andrade Dutra Filho

    2013-03-01

    Full Text Available Marcadores moleculares dos tipos RAPD e EST's SSR foram utilizados como ferramentas para avaliar a variabilidade bem como estimar a divergência genética entre variedades comerciais e clones de cana-de-açúcar oriundos via autofecundação. Vinte e três genótipos foram utilizados neste estudo oriundos do Programa de Melhoramento Genético da Cana-de-açúcar da Rede Interuniversitária para o Desenvolvimento do Setor Sucroalcooleiro (PMGCA/RIDESA. A extração do DNA genômico seguiu a metodologia CTAB com modificações para cana-de-açúcar. Foram utilizados 11 oligonucleotídeos RAPD da operon Technologies e 7 EST's SRR obtidos através de uma extensa revisão de literatura. As análises da divergência genética foram realizadas com o auxílio do Programa GENES. Os marcadores RAPD detectaram um alto grau de polimorfismo genético, produzindo 61 bandas, das quais 58 foram polimórficas. Os marcadores EST's SSR amplificaram 38 alelos, sendo 34 polimórficos. Havendo a formação de três grupos, com a população estudada. A maior parte da variação genética foi mantida dentro das progênies, evidenciando a ocorrência de um alto grau de variabilidade genética entre os genótipos de cada progênie para fins de melhoramento. Através da divergência genética estimada foi possível identificar parentais divergentes para trabalhos de hibridação, visando a obtenção de clones superiores com caracteres de interesse à agroindústria canavieira. Os marcadores molecuares RAPD e EST's SSR foram igualmente eficientes para estimar a variabilidade genética nos genótipos testados e elaborar os cruzamentos a serem realizados nos programas de melhoramento.Molecular markers of the type RAPD and ESTs SSR were used as tools to evaluate the variability, and estimate the genetic divergence between commercial varieties and sugarcane clones from self-pollination. Twenty-three genotypes from the Program for the Genetic Improvement of Sugarcane of the

  16. Total protein electrophoresis and RAPD fingerprinting analysis for the identification of Aeromonas at the species level Análise eletroforética de proteínas totais e marcadores de RAPD na identificação de Aeromonas ao nível de espécie

    Directory of Open Access Journals (Sweden)

    Ana Paula Longaray Delamare

    2002-12-01

    Full Text Available Fifteen well-defined strains of Aeromonas of thirteen species were analyzed by SDS protein electrophoretic analysis (SDS-PAGE and random amplified polymorphic DNA analysis (RAPD. The comparison between the patterns obtained by both methods allowed differentiating all the strains. Clusters formed by the unweighted pair group method with arithmetic averages applied to protein data correlates with the genetic and biochemical information about the species. The results show that protein fingerprinting has the potential to differentiate Aeromonas species, but the low qualitative variation indicates that this technique is not efficient for the characterization of strains within a species. Conversely, RAPD fingerprinting allows the identification of strains but the high variability limits its potential as an aiding method for species identification.Quinze linhagens de Aeromonas pertencentes a treze espécies foram avaliadas através de eletroforese de proteínas totais (SDS-PAGE e segmentos de DNA amplificados ao acaso (RAPD. A comparação entre os padrões obtidos por ambos métodos permitiu diferenciar todas as linhagens. Agrupamentos formados com base nos dados protéicos mostraram relação com informações bioquímicas e genéticas a respeito das espécies. Os resultados mostraram que análises protéicas têm potencial para diferenciar espécies de Aeromonas, mas a baixa variação qualitativa indica que esta técnica não é eficiente para a caracterização entre linhagens dentro de espécies. Ao contrário, marcadores de RAPD permitem identificar linhagens, mas a alta variabilidade limita seu potencial como método auxiliar na identificação de espécies.

  17. Assessment of genetic diversity on a sample of cocoa accessions resistant to witches' broom disease based on RAPD and pedigree data Avaliação da diversidade genética em uma amostra de acessos de cacau resistentes à doença vassoura-de-bruxa, com base em dados de RAPD e pedigree

    OpenAIRE

    Ronaldo Carvalho dos Santos; José Luís Pires; Uilson Vanderley Lopes; Karina Peres G. Gramacho; Acassi Batista Flores; Rita de Cássia S. Bahia; Helaine C. Cristine Ramos; Ronan Xavier Corrêa; Dario Ahnert

    2005-01-01

    Genetic diversity in cocoa (Theobroma cacao L.) has been assessed based on morphological and molecular markers for germplasm management and breeding purposes. Pedigree data is available in cocoa but it has not been used for assessing genetic relatedness. The geneitic diversity of 30 clonal cocoa accessions resistant to witche´ broom disease, from the CEPEC series, were studied on the basis of RAPD data and pedigree information. Twenty of these accessions descend from the TSA-644 clone, ...

  18. Genetic Variation in Rhizome Lotus (Nelumbo nucifera Gaertn. ssp. nucifera) Germplasms from China Assessed by RAPD Markers

    Institute of Scientific and Technical Information of China (English)

    AN Na; GUO Hong-bo; KE Wei-dong

    2009-01-01

    To estimate genetic variation in rhizome lotus (Nelumbo nucifera Gaertn. ssp. nucifera) gcrmplasms in China, a total of 94 rhizome lotus germplasms collected from 18 provinces in China were assessed. The RAPD (randomly amplified polymorphic DNA) marker was employed. The selected 17 random primers detected 139 polymorphic alleles out of a total 207 (67.15%). Nei's gene diversity statistics and region differentiation parameters indicated that all germplasms had a relatively high level of genetic diversity with ne=1.3202, h=0.1937, I=0.2982 and the gene flow among all regions was Nm=5.5742. The UPGMA dendrogram clustered all 94 germplasms into two clusters: One contained eight commercial cultivars and major landraces, and the other included the wild and some special landraces from five regions, and the PCA analysis exhibited the similar result. Those germplasms from southwestern and eastern China had higher genetic diversity than those from the southern, northern and central China. Predominant proportion of genetic variation (95.61%) was found significant within rather than among (4.39%) regions, as revealed by AMOVA analysis. The data analysis also revealed that the genetic diversity of rhizome lotus germplasms among different regions is positively related to their geographic distances, though it is ambiguous to find the trend from the UPGMA dendrogram and the PCA analysis. A relatively high genetic diversity and gene flow resided in the root lotus germplasms; about 96% of the variation was found within region; accessions from southwest and eastern China have higher genetic diversity than those from the southern, northern and central China.

  19. Characterization and genetic relatedness among 37 yardlong bean and cowpea accessions based on morphological characters and RAPD analysis

    Directory of Open Access Journals (Sweden)

    Vinich Saereeprasert

    2007-05-01

    Full Text Available Twenty four yardlong bean and 13 cowpea accessions were planted in the field to characterize their morphology and genetic relatedness. A randomized complete block design (RCBD with two replicationswas used. Growth habit, days to flowering, pod color, pod length, number of pods/plant, yield/plant and consumption quality were recorded. The results showed that pod length, number of pods/plant and podyield/plant among 37 accessions were highly significant differerence. Mean pod yield and pod length of 24 yardlong bean accessions were 212.1 g/plant and 48.7 cm, respectively, while mean pod yield and pod lengthof 13 cowpea accessions were 117.4 g/plant and 21.3 cm, respectively. Twenty two yardlong bean accessions exhibited indeterminate growth habit while 10 of 13 cowpea had determinate growth habit and the restsexhibited semi-determinate growth. Genetic variation and relationships among accessions were investigated based on RAPD technique. Total DNA was extracted from young leaf samples of all accessions using CTAB buffer. One hundred and twenty decamer oligonucleotide primers were screened and 5 primers (OPC-06,OPR-12, OPZ-03, OPZ-08, OPZ-13 were chosen for further evaluation. A dendrogram of genetic similarity was constructed based on 23 polymorphic bands obtained from 5 primers using UPGMA in SPSS program,which revealed separate groups between yardlong bean and cowpea. The similarity coefficient among yardlong bean and cowpea accessions ranged from 0.515 to 1.000 and 0.548 to 1.000, respectively.

  20. Identification of Herba Rabdosiae Serrae from Different Plant Resources by RAPD Method%应用RAPD技术对溪黄草基原植物分类鉴定

    Institute of Scientific and Technical Information of China (English)

    莫小路; 曾庆钱; 黄珊珊; 蔡岳文; 王玉生; 严振

    2012-01-01

    Objective:To identify the different plant resources of Herba Rabdosiae Serrae by using Random Amplified Polymorphic DNA ( RAPD ) Analysis. Methods: The Mini Spin Columns were used to extract the genomic DNA from five different plants of Herba Rabdosiae Serrae. With the DNA extracted from these plants as template,the 85 oligo nucleic acids (10 bp)as random primers,the Polymer Chain Reaction(PCR) was done and the results were analysed by electro-pharoses. Results: 12 primers were selected with polymorphism and 7 of them showed good polymorphism in RAPD map. Conclusion; RAPD method can be used to identify the plant resources of Herba Rabdosiae Serrae.%目的:用分子标记技术对中药溪黄草的基原植物分类鉴定.方法:用离心柱法提取5种溪黄草基原植物基因组DNA,用85个10 bp的寡核苷酸随机引物进行PCR(聚合酶链式反应),将扩增产物进行琼脂糖凝胶电泳,分析电泳图谱.结果:从85条随机引物中筛选出了12条多态性较好的引物,其中7个引物的扩增图谱表现较好的多态性,且重复性较好.结论:RAPD技术可以成功鉴定溪黄草药材基原.

  1. Molecular Diversity of Antagonistic Streptomyces spp. against Botrytis allii, the agent of onion gray mold using Random Amplified Polymorphic DNA (RAPD Markers

    Directory of Open Access Journals (Sweden)

    M. Jorjandi

    2014-08-01

    Full Text Available As an aim in sustainable agriculture, biological control of plant diseases has received intensive attention mainly as a response to public concern about the use of chemical fungicides in the environment. Soil Actinomycetes particularly Streptomyces spp. enhance soil fertility and have antagonistic activity against wide range of plant pathogens. To investigate for biocontrol means against the pathogen, 30 isolates of Actinomycetes have been isolated from agricultural soils of Kerman province of Iran and assayed for antagonistic activity against Botrytis allii, the agent of onion gray mold. RAPD DNA analysis has been used to determine the relatedness of active and non-active isolates based on their RAPD-PCR fingerprints. PCR amplifiable DNA samples have been isolated using the CTAB method and amplified fragments have been obtained from 5 random 10-mer primers. Different DNA fingerprinting patterns have been obtained for all of the isolates. Electrophoretic and cluster analysis of the amplification products has revealed incidence of polymorphism among the isolates. A total of 138 bands, ranging in size from 150-2800 bp, have been amplified from primers which 63.7% of the observed bands have been polymorphic. Genetic distances among different varieties have been analyzed with a UPGMA (Unweighted pair-group method, arithmetic average-derived dendrogram. Resulting dendrogram has showed from 0.65 to 0.91 similarities among varieties and divided the isolates into five major groups. Isolates which haven’t had any antagonistic activity against B. allii have been separated into a group and other isolates classified into four groups. The results indicate that RAPD is an efficient method for discriminating and studying genetic diversity of Streptomyces isolates.

  2. 使用RAPD分析辨别印度半岛的五种结鱼%Molecular discrimination of five Mahseer species from Indian peninsula using RAPD analysis

    Institute of Scientific and Technical Information of China (English)

    Vindhya MOHINDRA; Praveen KHARE; Kuldeep K.LAL; Peyush PUNIA; Rajeev K.SINGH; Anindya Sundar BARMAN; W.S.LAKRA

    2007-01-01

    用RAPD分析研究了结鱼种组(黄鳍结鱼Tor putitora, 结鱼Tor tor, 库德里结鱼Tor khudree, Tor mosal mahanadicus和墨脱四鬚魮Neolissochilus hexagonolepis)5个物种的遗传关系.在所测试的 69个随机引物中,11个引物能够在所有5个物种中扩增出稳定的条带.RAPD带型显示,综合使用这些RAPD标记能够区分这5个物种,但Tor mosal mahanadicus和黄鳍结鱼享有相似的带型.UPGMA分析揭示出3个独特的分支,第一支由黄鳍结鱼、Tor mosal mahanadicus和结鱼组成,第二支是库德里结鱼,第三支是墨脱四鬚魮.Tor mosal mahanadicus的分类地位在不同学者间存在分歧,被认为是库德里结鱼或结鱼的亚种,但我们的结果表明,相对而言,Mahanadi河中的Tor mosal mahanadicus与黄鳍结鱼的进化关系更近,因此有必要对其系统分类地位进行重新评估.%The genetic relatedness between five species of Mahseer group(Tor putitora, Tor tor, Tor khudree, Tor mosal mahanadicus and Neolissochilus hexagonolepis)was examined by Random Amplified Polymorphic DNA(RAPD)analysis for the first time. Out of the sixty-nine random primers tested, eleven primers generated scoreable patterns in all the five species. The comparative RAPD profiles revealed that the combination of RAPD markers could discriminate the species, except between T.mosal mahanadicus and T.putitora, which had a similar RAPD profile. UPGMA analysis depicted three distinct clusters; one formed by T.putitora, T.mosal mahanadicus and T.tor, the second by T.khudree and the third of Neolissochilus hexagonolepis. The taxonomic status of T.mosal mahanadicus is the subject of disagreement between authors; it has been considered as a subspecies of T.khudree and T.tor. The results demonstrated that the T.mosal mahanadicus of the river Mahanadi is closer to T.putitora than to the other Tor species studied and reassessment of its systematic position is required.

  3. Toxigenic diversity of two different RAPD groups of Stachybotrys chartarum isolates analyzed by potential for trichothecene production and for boar sperm cell motility inhibition

    DEFF Research Database (Denmark)

    Peltola, J.; Niessen, L.; Nielsen, Kristian Fog

    2002-01-01

    Thirty-one isolates of Stachybotrys chartarum from indoor and outdoor environments were analyzed for the presence of the trichodiene synthase (Tri5) gene, trichothecenes, boar sperm cell motility inhibition, and randomly amplified polymorphic DNA banding patterns (RAPDs). Twenty-two S. chartarum...... satratoxins or trichodermol. Nineteen S. chartarum isolates, distributed among the Tri5 gene negative and positive groups, inhibited boar spermatozoan motility at concentrations of less than or equal to60 mug of crude cell extract/mL. The inhibition of motility was independent of satratoxins or atranones...

  4. 重症监护室MRSA耐药性及RAPD分析%Antibiotics resistance and RAPD analysis of methicillin - resistant Staphylococcus aureus isolated from ICU

    Institute of Scientific and Technical Information of China (English)

    周秀萍; 杨长顺; 李争鸣; 邱宏

    2012-01-01

    Objective; To investigate the status of antibiotics resistance and the genetic homology of methicillin -resistant staphylococcus aureus in ICU and provide scientific evidence for controlling MRSA infection. Methods; MRSA was identified by mecA PCR. Drug resistance was perfomed using K - B method. The RAPD technique was used to identify the genotypes. Results; The separation rate of MRSA was 58. 5%. MRSA was resistant to many drugs. But all strains were susceptible to vancomycin and teicoplanin. All 24 stains of MRSA produced fingerprints by RAPD, and was classified into four genotypes. 15 strains were II type which was the dominant type. Conclusion; The MRSA isolated from ICU has high and multi drug resistance, we should reasonablly choose drugs according to the susceptibility test. RAPD results reveal the special epidemic genotypes of MRSA and this study can provide molecular epidemiological evidence for control of MRSA infection.%目的:调查重症监护室耐甲氧西林金黄色葡萄球菌的耐药及分子流行病学情况,为控制感染提供科学依据.方法:ICU分离的金黄色葡萄球菌采用mecA基因检测法确定MRSA,对筛选出的MRSA采用纸片法检测其耐药性,用随机扩增多态性DNA( RAPD)进行分子流行病学分析.结果:MRSA的分离率为58.5%,对多种抗生素均耐药,没有发现耐万古霉素和替考拉宁的菌株.24株MRSA经RAPD分型,均获得稳定的条带,可分为四型,以Ⅱ型为主,共检出15株.结论:ICU分离MRSA菌株具有多重耐药性,应根据药敏试验结果合理选用抗生素进行治疗;通过RAPD分型研究,可了解MRSA的基因型流行特征,为控制感染提供分子流行病学依据.

  5. OPTIMIZACIÓN DE LOS PROTOCOLOS DE EXTRACCIÓN DE ADN Y DEL MARCADOR MOLECULAR TIPO RAPD EN ANONÁCEAS

    OpenAIRE

    Yanet Alfonso Alonso; Caridad Noriega Carrera; Miriam Isidrón Pérez; Lucy Andraca Collazo; Dubiel Alfonso González; Daymara Rodríguez Alfonso

    2016-01-01

    Las técnicas moleculares requieren de protocolos que permitan determinar los niveles de variación genética, dentro de las poblaciones en diferentes condiciones ambientales. Tanto la optimización del aislamiento del ADN, como el de las condiciones de trabajo de las amplificaciones, son fundamentales para alcanzar el éxito de los análisis moleculares, por lo que la presente investigación tiene como objetivo: optimizar los protocolos de extracción de ADN y del marcador molecular tipo RAPD (Rando...

  6. Diversidade genética entre híbridos de laranja-doce e tangor 'Murcott' avaliada por fAFLP e RAPD Genetic diversity among hybrids of sweet orange and 'Murcott' tangor evaluated by fAFLP and RAPD markers

    Directory of Open Access Journals (Sweden)

    Marinês Bastianel

    2006-05-01

    Full Text Available O objetivo deste trabalho foi avaliar a diversidade genética em uma população de 148 híbridos de tangor 'Murcott' (Citrus reticulata Blanco x C. sinensis L. Osbeck e laranja 'Pêra' (C. sinensis L. Osbeck obtidos por polinização controlada, pelo uso de marcadores fAFLP e RAPD. Marcadores polimórficos (416 marcadores fAFLP e 33 RAPD foram utilizados para avaliar a similaridade genética entre os híbridos, calculada com o coeficiente Jaccard pelo método UPGMA. A consistência de cada agrupamento foi determinada pelo programa BOOD. Houve alta similaridade genética entre os parentais. A laranja 'Pêra' apresentou maior número (132 de loci em heterozigose em relação ao tangor 'Murcott' (105, corroborando a teoria de origem híbrida para a laranja-doce. Observaram-se dois grupos distintos de plantas, e um deles abrangeu 80% dos híbridos com maior similaridade com a laranja 'Pêra'. A análise bootstrap não revelou consistência estatística entre esses grupos. Marcadores fAFLP são mais eficientes na avaliação do polimorfismo, sendo indicados para seleção de indivíduos híbridos mais próximos a um dos parentais.The objective of this work was to evaluate the genetic diversity in a population of 148 hybrids of 'Murcott' tangor (Citrus reticulata Blanco x C. sinensis L. Osbeck and 'Pêra' sweet orange (C. sinensis L. Osbeck, obtained by controlled polination, using fAFLP and RAPD markers. Polymorphic markers (416 fAFLP and 33 RAPD markers were used to evaluate genetic similarity among the hybrids, calculated by the coefficient of Jaccard, using the UPGMA method. The consistency of each group was determined by software BOOD. There was high genetic similarity within the parents. 'Pêra' sweet orange had a higher number of loci in heterozygosis (132 compared to 'Murcott' tangor (105, supporting the theory of hybrid origin for sweet oranges. Two distinct groups of plants were observed: one group had 80% of the hybrids that displayed

  7. AL Amyloidosis

    Directory of Open Access Journals (Sweden)

    Desport Estelle

    2012-08-01

    Full Text Available Abstract Definition of the disease AL amyloidosis results from extra-cellular deposition of fibril-forming monoclonal immunoglobulin (Ig light chains (LC (most commonly of lambda isotype usually secreted by a small plasma cell clone. Most patients have evidence of isolated monoclonal gammopathy or smoldering myeloma, and the occurrence of AL amyloidosis in patients with symptomatic multiple myeloma or other B-cell lymphoproliferative disorders is unusual. The key event in the development of AL amyloidosis is the change in the secondary or tertiary structure of an abnormal monoclonal LC, which results in instable conformation. This conformational change is responsible for abnormal folding of the LC, rich in β leaves, which assemble into monomers that stack together to form amyloid fibrils. Epidemiology AL amyloidosis is the most common type of systemic amyloidois in developed countries with an estimated incidence of 9 cases/million inhabitant/year. The average age of diagnosed patients is 65 years and less than 10% of patients are under 50. Clinical description The clinical presentation is protean, because of the wide number of tissues or organs that may be affected. The most common presenting symptoms are asthenia and dyspnoea, which are poorly specific and may account for delayed diagnosis. Renal manifestations are the most frequent, affecting two thirds of patients at presentation. They are characterized by heavy proteinuria, with nephrotic syndrome and impaired renal function in half of the patients. Heart involvement, which is present at diagnosis in more than 50% of patients, leading to restrictive cardiopathy, is the most serious complication and engages prognosis. Diagnostic methods The diagnosis relies on pathological examination of an involved site showing Congo red-positive amyloid deposits, with typical apple-green birefringence under polarized light, that stain positive with an anti-LC antibody by immunohistochemistry and

  8. Curiosidad y mediación Del Lucio de Apuleyo al Anselmo/Lotario de Cervantes

    Directory of Open Access Journals (Sweden)

    Antonio Preciado Bernal

    1994-12-01

    Full Text Available Somos de la opinión de que El Curiosa lmpertinente (Torno II, Caps. 23, 24 y 25 no se puede analizar sin tener en cuenta la gran cornice constituida por la novela que lo rodea, Las aventuras del Ingenioso Hidalgo Don Quijote de la Mancha; no es menos cierto que este relato tiene una autonomía y se puede analizar también sin entrar en los paralelismos estructurales que indudablemente tiene con el todo de la obra. Estos paralelismos comienzan ya desde su colocación, no por azar simultánea en el tiempo de la narración ala lucha de don Quijote con los cueros de vino -clara referenda al episodio de Lucio y los odres de vino en el Asno de Oro de Apuleyo, la más elegante obra latina sobre la curiositas.

  9. KARAKTER GENETIK INDUK (F-0 DAN TURUNANNYA (F-1 PADA IKAN HIAS LAUT CLOWN (Amphiprion percula MENGGUNAKAN MARKER RAPD (Random Amplified Polymorfism DNA

    Directory of Open Access Journals (Sweden)

    Sari Budi Moria Sembiring

    2016-11-01

    Full Text Available Studi analisis karakter genetik ikan hias laut clown menggunakan metode penanda DNA RAPD dilakukan dalam upaya membantu pengembangan perbenihan dan budidaya ikan hias laut clown di Indonesia. Tujuan penelitian ini adalah untuk mendeterminasi karakter genetik dengan menggunakan analisis individu dari populasi induk (F-0 dan turunannya (F-1 sehingga diperoleh tingkat penurunan keragaman genetik dan keterkaitannya dengan karakter morfologi. Sampel yang dianalisis terdiri atas 5 pasang induk ikan clown (10 sampel dan masing-masing turunannya sebanyak 10 ekor (50 sampel sehingga total 60 sampel. Nilai rata-rata keragaman genetik induk ikan clown dari semua lokus primer sebesar 0,253, sedangkan pada turunannya (F-1 adalah 0,157. Hal ini menggambarkan adanya pengaruh genetik terhadap perbedaan pola pemunculan band putih. Study genetic characteristic of clownfish, Amphiprion percula using RAPD DNA marker was conducted in order to support development of breeding and culture program of marine ornamental clownfish in Indonesia. The objective of this research was to determine of genetic characteristic of clown fish using individual analysis from F-0 population and its generations (F-1 to find specific marker which is related to its morphology. Total samples analyzed were 60, consist of 5 pairs of clownfish broodstock (10 samples and 10 ind each generations (50 samples. Mean value of genetic diversity of clown fish broodstock from all primer loci was 0.253, while on its generation F-1 was 0.157. This result showed there was effect of genetic on the differences of white band pattern appearance.

  10. The population genetic structure of Littorina littorea (Mollusca: Gastropoda) along a pollution gradient in the Scheldt estuary (The Netherlands) using RAPD analysis.

    Science.gov (United States)

    De Wolf, Hans; Blust, Ronny; Backeljau, Thierry

    2004-06-05

    The population genetic structure of the periwinkle Littorina littorea was analysed using random amplified polymorphic DNA (RAPD). Three primers, coding for six putative polymorphic loci were surveyed to infer the genetic structure of seven populations located along the heavily polluted Western (i.e. in order of decreasing pollution load W1, W2, W3 and R1) and the relatively clean Eastern Scheldt (E1, E2 and E3) estuary (The Netherlands). A genetic distance based UPGMA (Unweighted pair group method with arithmetic mean) dendrogram revealed an estuary-related structuring, as Eastern and Western Scheldt sites formed two separate clusters. The Western Scheldt cluster was, however, much more heterogeneous, with three RAPD loci revealing a significant genetic heterogeneity compared to none when the Eastern Scheldt sites were compared. Overall mean heterozygosity levels were high, but did not reveal a difference between the estuaries. The current data (1) confirm the patterns of variation previously observed with electrophoretic analyses of esterases and (2) strongly support that these patterns of variation have a genetic basis, in the presence of intense gene flow. In addition, it is suggested that selection, rather than bottleneck effects, induced by the less favourable living conditions at W1, W2 and W3 are responsible for the genetic patterning.

  11. RAPDs na caracterização genético-molecular e no estudo da variabilidade genética de cultivares de ameixeira

    Directory of Open Access Journals (Sweden)

    Bianchi Valmor João

    2003-01-01

    Full Text Available Marcadores moleculares têm sido amplamente utilizados nas mais variadas espécies frutíferas para análise de "fingerprinting", para o processo de certificação de material vegetal e como ferramenta auxiliar em programas de melhoramento genético, para acessar a variabilidade genética entre genótipos. Dado a importância da cultura da ameixeira para a região Sul do Brasil, o presente trabalho teve por finalidade contribuir para a caracterização genético-molecular de 17 cultivares. As cultivares foram analisadas com 12 marcadores RAPD, que produziram 187 polimorfismos. O marcador OP A20 foi o mais polimórfico, produzindo 26 perfis diferentes. A análise de agrupamento, realizada com o método UPGMA, produziu um dendrograma que permitiu uma clara separação das cultivares em três grupos, correspondentes às suas respectivas espécies, Prunus salicina, Prunus domestica e Prunus cerasifera. O alto grau de polimorfismo detectado pelos marcadores RAPD confirma o potencial da técnica na análise de "fingerprinting" e sua utilidade na estimativa da variabilidade genética entre cultivares de ameixeira.

  12. Genetic variation of wild and hatchery populations of the catla Indian major carp (Catla catla Hamilton 1822: Cypriniformes, Cyprinidae revealed by RAPD markers

    Directory of Open Access Journals (Sweden)

    S.M. Zakiur Rahman

    2009-01-01

    Full Text Available Genetic variation is a key component for improving a stock through selective breeding programs. Randomly amplified polymorphic DNA (RAPD markers were used to assess genetic variation in three wild population of the catla carp (Catla catla Hamilton 1822 in the Halda, Jamuna and Padma rivers and one hatchery population in Bangladesh. Five decamer random primers were used to amplify RAPD markers from 30 fish from each population. Thirty of the 55 scorable bands were polymorphic, indicating some degree of genetic variation in all the populations. The proportion of polymorphic loci and gene diversity values reflected a relatively higher level of genetic variation in the Halda population. Sixteen of the 30 polymorphic loci showed a significant (p < 0.05, p < 0.01, p < 0.001 departure from homogeneity and the F ST values in the different populations indicated some degree of genetic differentiation in the population pairs. Estimated genetic distances between populations were directly correlated with geographical distances. The unweighted pair group method with averages (UPGMA dendrogram showed two clusters, the Halda population forming one cluster and the other populations the second cluster. Genetic variation of C. catla is a useful trait for developing a good management strategy for maintaining genetic quality of the species.

  13. Evaluation of genetic variability in micropropagated propagules of ornamental pineapple [Ananas comosus var. bracteatus (Lindley) Coppens and Leal] using RAPD markers.

    Science.gov (United States)

    Santos, M D M; Buso, G C S; Torres, A C

    2008-10-21

    The objective of the present study was to evaluate the genetic variability in micropropagated plantlets of ornamental pineapple, after the fourth period of subculture. The basal culture medium consisted of MS salts, vitamins, 3% sucrose, liquid formulation, supplemented with 6-benzylaminopurine (BAP) at concentrations of 0.125, 0.25, 0.5, 1.0, and 2.0 mg/L. The addition of BAP influenced the occurrence of genetic variation revealed using random amplified polymorphic DNA (RAPD) markers. Of a total of 520 primers tested, 44 were selected and amplified; 402 monomorphic bands (97.2%) and 18 polymorphic bands (2.8%) resulted among regenerated plantlets. The polymorphic fragments were produced by 12 primers (OPA-01, OPA-20, OPB-01, OPB-19, OPC-19, OPF-13, OPL-17, OPM-13, OPP-16, OPT-07, OPV-19, and OPX-03). Among the primers that identified polymorphism, OPA-01, OPA-20, OPB-19, OPC-19, OPL-17, OPP-16, and OPX-3 each showed, one polymorphic band and OPF-13 amplified a maximum of three bands. In this study, the RAPD technique was effective in showing the occurrence of somaclonal variations that occur during the micropropagation process of ornamental pineapple cultivation in BAP-supplemented medium, and it is possible to detect the presence of genetic variation in early stages of plant development.

  14. A genetic map of cucumber composed of RAPDs, RFLPs, AFLPs, and loci conditioning resistance to papaya ringspot and zucchini yellow mosaic viruses.

    Science.gov (United States)

    Park, Y H; Sensoy, S; Wye, C; Antonise, R; Peleman, J; Havey, M J

    2000-12-01

    The watermelon strain of papaya ringspot virus (PRSV-W) and zucchini yellow mosaic virus (ZYMV) are potyviruses that cause significant disease losses in cucumber. Resistances have been identified primarily in exotic germplasm that require transfer to elite cultivated backgrounds. To select more efficiently for virus resistances, we identified molecular markers tightly linked to PRSV-W and ZYMV resistances in cucumber. We generated F6 recombinant inbred lines (RILs) from a cross between Cucumis sativus L. 'Straight 8' and a line from 'Taichung Mou Gua', TMG1 (susceptible and resistant, respectively, to both viruses), and studied the segregations of amplified fragment length polymorphism (AFLP) markers, randomly amplified polymorphic DNAs (RAPDs), restriction fragment length polymorphisms (RFLPs), and resistances to PRSV-W and ZYMV. A 353-point map of cucumber was generated, delineating 12 linkage groups at LOD 3.5. Linkage arrangements among RFLPs were consistent with previously published maps; however linkages among RAPDs in our map did not agree with a previously published map. Resistances to PRSV-W and ZYMV were tightly linked (2.2 cM) and mapped to the end of one linkage group. One AFLP cosegregated with resistance to ZYMV.

  15. Identification and characterization of RAPD-SCAR markers linked to glyphosate-susceptible and -resistant biotypes of Eleusine indica (L.) Gaertn.

    Science.gov (United States)

    Cha, Thye San; Anne-Marie, Kaben; Chuah, Tse Seng

    2014-02-01

    Eleusine indica is one of the most common weed species found in agricultural land worldwide. Although herbicide-glyphosate provides good control of the weed, its frequent uses has led to abundant reported cases of resistance. Hence, the development of genetic markers for quick detection of glyphosate-resistance in E. indica population is imperative for the control and management of the weed. In this study, a total of 14 specific random amplified polymorphic DNA (RAPD) markers were identified and two of the markers, namely S4R727 and S26R6976 were further sequence characterized. Sequence alignment revealed that marker S4R727 showing a 12-bp nucleotides deletion in resistant biotypes, while marker S26R6976 contained a 167-bp nucleotides insertion in the resistant biotypes. Based on these sequence differences, three pairs of new sequence characterized amplified region (SCAR) primers were developed. The specificity of these primer pairs were further validated with genomic DNA extracted from ten individual plants of one glyphosate-susceptible and five glyphosate-resistant (R2, R4, R6, R8 and R11) populations. The resulting RAPD-SCAR markers provided the basis for assessing genetic diversity between glyphosate-susceptible and -resistant E. indica biotypes, as well for the identification of genetic locus link to glyphosate-resistance event in the species.

  16. Optimization of DNA isolation and PCR protocol for analysis and evaluation of genetic diversity of the medicinal plant, Anemopsis californica using RAPD.

    Science.gov (United States)

    Lizette Del-Toro-Sánchez, C; Villaseñor-Alvarado, S; Zurita-Martínez, Florentina; Castellanos-Hernández, O A; Rodríguez-Sahagún, Araceli; Isabel Torres-Morán, M; Rojas-Bravo, D; Gutiérrez-Lomelí, M

    2013-06-01

    Anemopsis californica is a perennial herbaceous plant that has been utilized as a medicinal plant for the treatment of various diseases. The present work was carried out with the objective of optimizing a method of extraction of the genomic DNA of A. californica and a PCR protocol and later to evaluate the existing genetic diversity among the genotypes deriving from different origins. For DNA extraction, we tested four procedures: with the CTA B-2 protocol, we obtained the highest yield (61.5±2.2 μg DNA/g of leaf tissues) and the best quality (A260/280 1.83±0.022). To estimate genetic variability, we utilized the randomly amplified polymorphism DNA (RAPD) technique, employing 20 oligonucleotides, of which only 18 generated reproducible banding patterns, producing 123 polymorphic bands generated, thus obtaining a polymorphism rate of 93.93% among the genotypes analyzed. The Jaccard similarity coefficient generated a variation ranging from 0.325-0.921, indicating a high level of genetic variation among the studied genotypes. An Unweighted pair-group method with arithmetic mean (UPGMA) group analysis indicated six distinct groups. The present optimized method for DNA isolation and RAPD protocol may serve as an efficient tool for further molecular studies.

  17. Characterization of RAPD Markers, and the RFLP Marker Linked to Powdery Mildew Resistance Gene Derived from Different Accessions of H. villosa

    Institute of Scientific and Technical Information of China (English)

    LI Hui; CHEN Xiao; SHI Ai-nong; KONG Fan-jing; S Leath; J P Murphy; JIA Xu

    2005-01-01

    The analysis was carried out on performance of the resistance gene from Haynaldia villosa accession of the former Soviet Union to different isolates of Bluemerie graminis. Polymorphisms were revealed between 6D/6V substitution line Pm930640and its pedigree parents using five RAPD markers of OPAN031700, OPAI017oo, OPAL03750, OPAD07480 and OPAG1558oscreened out from 120 random 10-mers primers. Three RAPD markers of OPAN03, OPAI01 and OPAL03 were linked with the resistance gene by analysis of F2 population of Chancellor×Pm930640. Analysis of 29 wheat lines including part of lines conferring the known genes from Pm1 to Pm20 respectively, lines conferring resistance gene from two H. villosaaccessions and the related wheat parents, were analyzed and the results showed that these markers not only linked to thegene resistant to powdery mildew from H. villosa, but also detected different genetic backgrounds. OPAL03750 can beused as the marker to distinguish the different resistant lines from two H. villosa accessions because it was only observedin the materials from H. villosa of the former Soviet Union. RFLP analysis also showed the polymorphisms between twoH. villosa accessions and their derived resistant lines.

  18. RAPD Analysis on the Relationship among the Diploid Species in Aegilops%山羊草属二倍体物种亲缘关系的RAPD分析

    Institute of Scientific and Technical Information of China (English)

    蔡从利; 王建波; 朱英国

    2001-01-01

    利用24个随机引物对山羊草属12个二倍体物种的亲缘关系进行了RAPD分析,对扩增出的304条带进行聚类分析。结果发现:(1)Sitopsis组内各物种的亲缘关系与前人的研究基本一致;(2)Ae.mutica与Comopyrum组的各物种亲缘关系较近;(3)Ae .uniaristata与同组的另两个物种亲缘关系较远,应将其从该组中独立出来;(4)Ae. caudata与Ae.umbellulata为亲缘关系较近的两个物种。%RAPD analysis was performed by a set of 24 arbitrary primers to 12 diploid species in Aegilops. Results were observed based on t he examination of 304 RAPD fragments. The relationship in the Section Sitopsis i s consistent with the former investigation. Ae. mutica had a relatively clos e relationship with the species of the Section Comopyrum.Ae. uniaristata had a far relationship with the other two species of the Section Comopyrum,indicati ng that it should be excluded from this section. Ae. caudata and Ae. umbel lulata had a close relationship.

  19. Caracterización, por RAPD-PCR, de aislados de Pseudomonas aeruginosa obtenidos de pacientes con fibrosis quística RAPD-PCR characterization of Pseudomonas aeruginosa strains obtained from cystic fibrosis patients

    Directory of Open Access Journals (Sweden)

    Maribel Ortiz-Herrera

    2004-04-01

    Full Text Available OBJETIVO: Caracterizar a las cepas de P aeruginosa aisladas de lavados broncoalveolares de pacientes con fibrosis quística a lo largo de un periodo de tres años. MATERIAL Y MÉTODOS: Estudio prospectivo, de seguimiento de una población de pacientes con fibrosis quística. Se utilizó la técnica de la amplificación del ADN empleando PCR con bajas condiciones de especificidad (Random amplified polymorphic DNA, RAPD-PCR para la amplificación del ADN de cepas de P aeruginosa aisladas de lavados broncoalveolares de cinco pacientes con fibrosis quística, provenientes del Servicio de Neumología y Cirugía del Tórax del Instituto Nacional de Pediatría de la Ciudad de México, en el periodo de junio de 1996 a junio de 2002; se establecieron los patrones de amplificación de cada aislamiento, lo que permitió la identificación precisa de todas las cepas aisladas y el estudio de la epidemiología de P aeruginosa en los pacientes seleccionados con dicha enfermedad. RESULTADOS: Se definieron 18 patrones de amplificación del ADN que permitieron identificar a cada cepa de P aeruginosa aislada en las diferentes muestras de lavado broncoalveolar; no se encontró relación entre el fenotipo de P aeruginosa (mucoide o no mucoide y el genotipo de cada aislamiento, ya que cepas con fenotipos distintos mostraron patrones de amplificación semejantes; en nuestros pacientes se identificaron cepas con patrones de amplificación distintos a partir de una misma muestra, lo que sugiere la presencia de infecciones simultáneas por más de una cepa de P aeruginosa; se demostró que dos hermanos con la enfermedad compartían cepas con genotipos semejantes, lo que sugiere una contaminación cruzada entre ambos, y se demostró el aislamiento de cepas de P aeruginosa con genotipos semejantes a lo largo de los periodos estudiados. CONCLUSIONES: La identificación mediante la caracterización genotípica de las cepas de P aeruginosa aisladas de los pacientes con

  20. 玳瑁遗传多样性的RAPD分析%Genetic Diversity Analysis of Eretmochelys Imbricata by RAPD Method

    Institute of Scientific and Technical Information of China (English)

    端金霞; 古河祥; 夏中荣; 叶明彬; 陈华灵; 张飞燕

    2011-01-01

    We used RAPD software to analyze the genetic diversity of seven Hawksbill Turtles(Eretmochelys imbricata) captured in the South China Sea.We used 20 random primers to amplify 1351 fragments.On average,193 bands were amplified for each individual.Sixty - nine of 193 loci were polymorphic(35.8%).The size of DNA fragments ranged from 200 bp to 3 000 bp.The genetic distance among individuals ranged from 0.082 9 to 0.181 3 and averaged 0.132 7±0.029 9.The phylogenetic tree of the seven turtles was constructed by NJTREE analysis using the RAPDistance 1.04 program.We conclude that the genetic diversity of Eretmochelys imbricata in the South China Sea is low.%应用RAPD技术分析了玳瑁的遗传多样性。用20个随机引物对中国南海海域玳瑁7个个体的基因组DNA进行了PCR扩增,共扩增出1 351条DNA片段,平均每个个体扩增出193条条带。在检测到的193条条带中,多态性条带为69条,多态性条带百分比为35.8%,条带大小在200 bp~3 000 bp之间,7个个体间遗传距离为0.082 9~0.1813,平均遗传距离为0.132 7±0.029 9,表明中国南海海域玳瑁的遗传多样性水平较低,应加强该区域玳瑁种质资源的保护。采用类平均聚类法(NJTREE)构建了7个个体相互关系的分子聚类图,表明该7个玳瑁个体没有形成种群的分化。

  1. Impacto de los programas de movilidad internacional en la adquisición de competencias académicas para el ingreso al mercado laboral: México

    Directory of Open Access Journals (Sweden)

    Elvira Ivone González Jaimes

    2016-06-01

    Full Text Available El objetivo es realizar un análisis de las competencias académicas que influyen en la adquisición y conservación del empleo de los estudiantes y egresados de una universidad estatal. Dichas variables fueron estudiadas a través de realizar un análisis comparativo entre estudiantes y egresados con y sin participación en el Programa de Movilidad Universitaria Internacional (PMUI. El método utilizado es de diseño no experimental, comparativo en formato de encuesta, corte transversal, selección de la muestra al azar (estudiantes o egresados con y sin movilidad internacional como empleadores para realizar un análisis estadístico de tipo descriptivo e inferencial. Las poblaciones son: 1 391 estudiantes y egresados, 68 empleadores. Muestras al azar: 372 estudiantes y egresados como 54 empleadores. Material: encuesta electrónica del cuestionario de competencias para estudiantes, egresados y empleadores.

  2. Epidemiology and clinical feature of kala azar: Analysis of 47 cases%黑热病47例流行病学及临床特征分析

    Institute of Scientific and Technical Information of China (English)

    陈安弟; 余珍; 游敏; 黄丽; 胥雪艳

    2011-01-01

    To discuss the epidemiology and clinical features of kala azar to provide evidence for reducing misdiagnosis and mistreatment Methods We used bone marrow aspiration or rk39 test paper strip to test the patients. Results A total 47 patients had the history of living in the epidemic areas. The disease attacked monthly. The most frequent onset time was from Feb to Jun (68. 09% ). There were more male patients than female patients. The age of onset was manly less than 16. The ones under 16 years old accounted for 80. 85% of the patients. About 95. 75% patients took 6 months from onset to final diagnosis. The course of disease was related to the time of misdiagnosis. The main clinical manifestations were the long-term irregular fever (100. 00% ) , progressive a-nemia (100% ), splenomegaly (95. 74% ) pancytopenia {89. 63% ) , and the increase of plasma protein (57.45% ). The incidence of masculine of the rk39 dipstick check was 100% , which was significantly higher than that of bone marrow aspiration smear (55. 56% ). Conclusions It is of critical importance to understand the epidemiology and clinical features of kala azar and carry out timely check of pathogen and blood serum.%目的 探讨黑热病的流行病学与临床特征,为减少误诊误治提供依据.方法 对就诊患者采用骨髓穿刺或rk39试纸条检测,阳性者收集整理流行病学、发病情况、临床表现、辅助检查和病历记录等资料,给予锑剂药物治疗.结果47例患者均有流行区居住史;各月均有发病,以2~6月居多,占全年发病的68.09%;男性多于女性,年龄以16岁及以下发病较多,占发病数的80.85%;发病至确诊时间6个月内的占95.75%,病程长短与误诊时间有关.主要临床表现为长期不规则发热( 100.00%),进行性贫血和脾肿大(100.00%、95.74%);以白细胞为主的全血细胞减少及血浆球蛋白增高(89.63%和57.45%).47例rk39试纸条检查阳性为100%,

  3. Utility of RAPD marker for genetic diversity analysis in gamma rays and ethyl methane sulphonate (EMS)-treated Jatropha curcas plants.

    Science.gov (United States)

    Dhakshanamoorthy, Dharman; Selvaraj, Radhakrishnan; Chidambaram, Alagappan

    2015-02-01

    The presence of important chemical and physical properties in Jatropha curcas makes it a valuable raw material for numerous industrial applications, including the production of biofuel. Hence, the researcher's interest is diversified to develop more and better varieties with outstanding agronomic characteristics using conventional breeding. Among these, mutation breeding is one of the best approaches to bring genetic changes in plant species. The aim of this study is to evaluate the diversity and genetic relationship among J. curcas mutants, which were obtained from different doses of gamma rays (control, 5 Kr, 10 Kr, 15 Kr, 20 Kr and 25 Kr) and EMS (1%, 2%, 3% and 4%), using RAPD marker. Among the 21 random primers, 20 produced polymorphic bands. The primers, OPM-14 and OPAW-13, produced a minimum number of bands (3) each across the ten mutants, while the primer OPF-13 produced the maximum number of bands (10), followed by the primers OPU-13, OPAM-06, OPAW-09 and OPD-05, which produced 9 bands each. The number of amplicons varied from 3 to 10, with an average of 7 bands, out of which 4.57 were polymorphic. The percentage of polymorphism ranged from 0.00 to 100 with an average of 57%. In the present study, RAPD markers were found most polymorphic, with an average polymorphism information content (PIC) value of 0.347, effective multiplex ratio (EMR) of 35.14, marker index (MI) of 14.19, resolution power (Rp) of 11.19, effective marker index (EMI) of 8.21 and genotype index (GI) of 0.36, indicating that random primers are useful in studies of genetic characterization in J. curcas mutant plants. In a dendrogram constructed based on Jaccard's similarity coefficients, the mutants were grouped into three main clusters viz., (a) control, 10 Kr, 15 Kr, 20 Kr, 2% EMS, and 3% EMS, (b) 5 Kr and 1% EMS, and (c) 25 Kr and 4% EMS mutants. Based on the attributes of the random primers and polymorphism studied, it is concluded that RAPD analysis offers a useful molecular marker

  4. New method for early detection of two random amplified polymorphic DNA (RAPD groups of Staphylococcus aureus causing bovine mastitis infection in Paraná State, Brazil

    Directory of Open Access Journals (Sweden)

    Dicezar Gonçalves

    2010-04-01

    Full Text Available The aim of this work was to develop a fast and accurate molecular approach to allow early detection of two RAPD groups of S. aureus causing bovine mastitis. Seventy five S. aureus isolates from infected animals were characterized by RAPD. Genomic fragments isolated from the unique bands present in either group were cloned and sequenced. Based on the DNA sequences, specific primers were designed to allow for the simultaneous detection of either group by multiplex PCR of S. aureus DNA isolated from clinical and subclinical bovine mastitis. Results showed that these proposed primers set could be used to detect various clinical and subclinical S. aureus isolates as well as the detection of the microorganism in bulk milk. Their use as a specific method for effective and early diagnostic tool for S. aureus infection in dairy herds is suggested.Esta pesquisa objetivou o desenvolvimento de técnica rápida e eficiente para diagnosticar precocemente diferentes linhagens de S. aureus causadoras de mastite bovina. Como resultados da metodologia empregada, foram isoladas duas linhagens destas bactérias que causam diferentes tipos de mastite bovina. Os fragmentos de DNA genômico caracterizando ambas as linhagens, por meio de RAPD foram inseridos em vetor plasmidial pGEM e clonados por meio de clones T10 F1 de Escherichia coli. As seqüências obtidas permitiram desenhar iniciadores específicos para o reconhecimento de ambas as linhagens, os quais foram testados com amostras de S. aureus e com outras linhagens próximas. O diagnóstico por meios moleculares, pode ser realizado diretamente de amostras coletadas de rebanhos leiteiros assim como dos equipamentos de ordenha. A significância deste estudo consiste em um rápido e acurado método para localizar animais infectados, representando importante ferramenta no manejo do rebanho, na redução de custos com tratamentos e, rápida recuperação de rebanhos infectados.

  5. Diversidade genética de três estoques de piapara (Leporinus elongatus, utilizando RAPD - DOI: 10.4025/actascianimsci.v30i2.4710 Genetic diversity of three stocks of piapara (Leporinus elongatus, using RAPD - DOI: 10.4025/actascianimsci.v30i2.4710

    Directory of Open Access Journals (Sweden)

    Lauro Vargas

    2008-08-01

    Full Text Available Recentemente a produção aquícola brasileira tem apresentado grande progresso. Dentre as espécies nativas cultivadas no Brasil, a piapara (Leporinus elongatus tem sido amplamente preconizada. Com objetivo de avaliar os programas de repovoamento, foram analisadas a variabilidade e a divergência genética de três estoques de piapara com a técnica de RAPD (Random Amplified Polymorphic. O primeiro estoque pertence à Estação de Aquicultura e Hidrologia da Duke Energy International (A; o segundo, à piscicultura de Rolândia (B e o terceiro, ao Programa de Repovoamento dos Rios do Paraná (C. Os dez primers para RAPD utilizados produziram 105 fragmentos polimórficos, conferindo um polimorfismo de 98,1% para os três estoques avaliados. A porcentagem de locos polimórficos e índice de Shannon foi superior para o estoque A. Porém, todos valores foram elevados, indicando alta diversidade intrapopulacional. Os valores de indicam que houve baixa diferenciação genética entre os estoques A x B e moderada diferenciação entre os demais. O Nm foi maior entre os estoques A x B. A distância genética e o dendrograma indicam que os estoques A x B são menos distantes geneticamente.Latelly, aquiculture production in Brazil has made great strides. Among the native species cultivated in Brazil, piapara (Leporinus elogatus has been widely praised. With the objective of evaluating restocking programs, the variability and genetic divergence of three piapara stocks were analyzed using the RAPD (Random Amplified Polymorphic DNA technique. The first stock belongs to the Aquiculture and Hydrology Station of Duke Energy International (A; the second one belongs to a fish farm in the city of Rolândia (B; and the third to the River Restocking Program of Paraná (C. The ten primers used for RAPD produced 105 polymorphic loci, conferring a polymorphism of 98.1% for the three evaluated stocks. Polymorphic loci percentage and Shannon index were higher for stock A

  6. Genotype characterization of Haematobia irritans from different Brazilian geographic regions based on randomly amplified polymorphic DNA (RAPD analysis Caracterização genotípica de Haematobia irritans procedentes de diferentes regiões geográficas brasileiras baseada na análise do DNA polimórfico amplificado ao acaso (RAPD-PCR

    Directory of Open Access Journals (Sweden)

    Luciana G. Brito

    2007-01-01

    Full Text Available Blood-sucking diptera are important parasites in bovine production systems, especially regarding confinement conditions. Haematobia irritans, the horn fly, is one of the most troublesome species within bovine production systems, due to the intense stress imposed to the animals. An important aspect while studying the variability within a species is the study of the geographic structure of its populations and, attempting to find out the genetic flow of Brazilian populations of horn fly, the RAPD technique, which is suited for this purpose, has been used. The use of molecular markers generated from RAPD made it possible to identify the geographic origin of samples from different Brazilian geographic regions, as well as to estimate the genotypic flow among the different Brazilian populations of the horn fly.Dípteras hematófagos são importantes parasitas dentro de sistemas de produção de bovinos, especialmente em confinamento. Haematobia irritans, a mosca-dos-chifres, é uma das espécies que maiores problemas causa em sistemas de produção de bovinos, dado ao intenso estresse que impõe aos animais. Um importante aspecto quando se estuda a variabilidade genética dentro das espécies é o estudo da estrutura geográfica destas populações. Buscando-se estimar a similaridade genotípica das diferentes populações brasileiras da mosca do chifre utilizou-se a técnica do DNA polimórfico amplificado ao acaso (RAPD-PCR, que mostrou-se eficiente para tal propósito. A utilização dos marcadores moleculares gerados através da técnica de RAPD-PCR tornou possível a identificação da origem geográfica das amostras das diferentes regiões geográficas brasileiras, assim como, estimar o fluxo genotípico entre as diferentes populações brasileiras da mosca-dos-chifres.

  7. Marcadores moleculares RAPD e descritores morfológicos na avaliação da diversidade genética de goiabeiras (Psidium guajava L. = RAPD molecular markers and morphological descriptors in the evaluation of genetic diversity of guava (Psidium guajava L.

    Directory of Open Access Journals (Sweden)

    Aroldo Gomes Filho

    2010-10-01

    Full Text Available O conhecimento da variabilidade genética e fenotípica entre diferentes acessos de goiabeiras é importante para se apoiar programas de melhoramento dessa espécie na região Norte Fluminense que carece de novas culturas capazes de gerar renda aos produtores locais. O objetivo deste trabalho foi avaliar a divergência genética entre seis cultivares e 19 acessos de goiabeiras, por meio de marcadores moleculares RAPD e características morfoagronômicas. Foram obtidas 117 marcas polimórficas, utilizando-se 28 iniciadores. Os resultados mostraram uma concordância parcial entre os métodos de agrupamentos estudados, com a formação de 12 grupos. O acesso Vita 3 e o acesso 6 foram os mais divergentes, apresentando distância genética de 0,663. A análise comparativa dos agrupamentos revelou que os marcadores RAPD e os descritores morfológicos foram eficientes para discriminação dos acessos e que houve variabilidade genética potencial para uso em Programa de Melhoramento Genético.The knowledge of the genetic and phenotypic variability among different accessions of guava is important for supporting improvement programs of this specie in northern Rio de Janeiro state, which needs new cultivars able to generate income for local farmers. This work aimed to evaluate the genetic divergence among six cultivars and 19 accessions of guava via RAPD molecular markers and morphologicalcharacteristics. One hundred and seventeen polymorphic markers were obtained from 28 primers. The results showed a partial agreement between the methods of studied groupings, with the formation of 12 groups. The accessions ‘Vita 3’and ‘6’ were the most divergent, showing genetic distance of 0.663. The comparative analysis of groupings showed that RAPD markers and morphological descriptors were effective in discriminating the accessions and to show potentialgenetic variability useful in genetic improvement programs.

  8. Molecular heterogeneity of Malassezia pachydermatis through RAPD-PCR - doi: 10.4025/actascibiolsci.v32i2.4788 Molecular heterogeneity of Malassezia pachydermatis through RAPD-PCR - doi: 10.4025/actascibiolsci.v32i2.4788

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    João Roberto Braga Mello

    2010-05-01

    Full Text Available Several methodologies in molecular biology have been used in the investigation of Malassezia pachydermatis and its differentiation into subtypes. Recent molecular research of this species includes the use of samples isolated from canine otitis externa and dermatitis, as well as from healthy animals, having in view an epidemiological study of the yeast. The aim of this study was to identify molecular differences in M. pachydermatis samples isolated from dogs with otitis externa. The M. pachydermatis strains were analyzed by means of the Random Amplification Primer DNA - Polimerase Chain Reaction (RAPD–PCR for molecular heterogeneity research. DNA extraction was carried out with phenol-chloroform and the RAPD technique using the AGAATCCGCC primer. A variation was observed in the number and arrangement of the bands among the 49 studied isolates, grouped into nine patterns. Isolate groupings were not found to be related to animal breed, age or sex. It was concluded that M. pachydermatis has differences in its molecular profile, as shown by the molecular technique (RAPD – PCR, which allows isolates to be classified into nine subtypes.Several methodologies in molecular biology have been used in the investigation of Malassezia pachydermatis and its differentiation into subtypes. Recent molecular research of this species includes the use of samples isolated from canine otitis externa and dermatitis, as well as from healthy animals, having in view an epidemiological study of the yeast. The aim of this study was to identify molecular differences in M. pachydermatis samples isolated from dogs with otitis externa. The M. pachydermatis strains were analyzed by means of the Random Amplification Primer DNA - Polimerase Chain Reaction (RAPD–PCR for molecular heterogeneity research. DNA extraction was carried out with phenol-chloroform and the RAPD technique using the AGAATCCGCC primer. A variation was observed in the number and arrangement of the bands

  9. Análisis de la variación genética en clones de caucho (Hevea brasiliensis de Asia, Suramérica y Centroamérica usando marcadores RAPD Analysis of genetic variation in clones of rubber (Hevea brasiliensis from Asían, South and Central American origin using RAPDs markers

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    Lobo Arias Mario

    2006-12-01

    Full Text Available El caucho natural (Hevea brasiliensis representa a especies potenciales para reforestación y programas de explo­tación comercial en ciudades tropicales como Colombia. La variabilidad genética de una colección de caucho que se encuentra en la Estación experimental de Paraguaycito en Buenavista, departamento del Quindio en Colom­bia fue estudiada para aumentar el conocimiento en cuanto a las especies y realizar un mejor uso de los árboles disponibles. Un total de 25 clones, seis de Sur América, 17 de Asia y 2 de América Central fueron seleccionados y analizados usando RAPDs. Las muestras aisladas de ADN de los árboles fueron con 102 primers, 23 de los cuales mostraron polimorfismos. Aunque se encontró un alto grado de similaridad, los análisis grupales de datos llevaron a diferenciar los árboles de de caucho en términos de su origen geográfico. Por lo tanto, las relaciones genéticas que se encontraron entre los clones podrían ayudar a seleccionar parentales para uso en programas de reproducción y diseño de estrategias para la conservación de los clones que tengan características agronómicas deseables. Palabras clave: identificación de cultivos, distancia genética, diversidad genética, Hevea brasiliensis, marcadores RAPD.Rubber (Hevea brasiliensis represents a potential species for reforestation and commercial exploitation programmes in tropical countries such as Colombia. The genetic variability of a rubber collection kept at the Paraguaicito Experimental Station in Buenavista in the Quindio department of Colombia was studied to improve knowledge regarding this species and make better use of the trees available. A total of 25 clones, six from South-America, 17 from Asia and two from Central-América were selected and analysed using RAPDs. DNA samples isolated from the trees were screened with 102 primers, 23 of which revealed polymorphism. Although a high degree of similarity was found, clustering analysis of the data led

  10. Development of SCAR Markers Based on Improved RAPD Amplification Fragments and Molecular Cloning for Authentication of Herbal Medicines Angelica sinensis, Angelica acutiloba and Levisticum officinale.

    Science.gov (United States)

    Zhang, Chun; Mei, Zhiqiang; Cheng, Jingliang; He, Yin; Khan, Md Asaduzzaman; Luo, Peiyi; Imani, Saber; Fu, Junjiang

    2015-10-01

    Molecular cloning from DNA fragments of improved RAPD amplification of Angelica sinensis, Angelica acutiloba and Levisticum officinale, provided novel sequence-characterized amplified region (SCAR) markers A13, A23, A1-34 and A1-0 whose sequences were deposited in the GenBank database with the accession numbers KP641315, KP641316, KP641317 and KP641318, respectively. By optional PCR amplification, the SCAR markers A13 and A23 are Levisticum officinale-specific, whereas the SCAR marker A1-34 is Angelica acutiloba-specific, and the SCAR marker A1-0 is Angelica sinensis-specific. These diagnostic SCAR markers may be useful for genetic authentications, for ecological conservation of all three medicinal plants and as a helpful tool for the genetic authentication of adulterant samples.

  11. RAPD fingerprinting: use in the analysis of mediterranean populations of European fallow deer, Dama dama Linnaeus, 1758 (Mammalia, Artiodactyla

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    Marco Masseti

    1996-12-01

    Full Text Available Abstract The aim of the present paper is to present a preliminary genetic survey of the fallow deer (Dama dama L., 1758 population of the island of Rhodes (Greece in order to verify its genetic variability. Italian population specimens were chosen as a control group because, as can be ascertained from literature, they have a very low level of variability. The analysis was carried out on hair samples obtained from each population. It was performed on a PCR modified method called RAPD which is based on the amplification of genomic DNA by using a single oligonucleotide of random sequence as a primer. The results of the analysis of the Rhodian specimens show clearly the presence of polymorphic individuals, absent in the Italian deer sampled.

  12. Assessment of air pollution genotoxicity by RAPD in Evernia prunastri L. Ach. from around iron-steel factory in Karabük, Turkey.

    Science.gov (United States)

    Cansaran-Duman, Demet; Atakol, Orhan; Aras, Sümer

    2011-01-01

    Lichens are widely used in biomonitoring studies of air pollution, either as bioindicators of air quality or as bioaccumulators of atmospheric deposition. Over the past decade, several molecular techniques have been developed to provide information on diversity, genotoxicology, genetic relationships, etc. The heavy metal contents of Evernia prunastri samples were determined by atomic absorption spectrometry. The Random Amplified Polymorphic DNA Polymerase Chain Reaction (RAPD-PCR) method was used to describe the pattern of DNA band variation in the samples influenced by the environmental pollution. The study was designed to describe the level of pollution in an area contaminated with smoke and waste from an iron-steel factory, and to reveal the level of potential genotoxic agents around this source of pollution. The study also examined the suitability of the lichen samples for the detection of genotoxicity.

  13. 片形吸虫DNA随机扩增多态性分析%Detection of Difference Among Fasciola hepatica and Fasciola gigantica with RAPD

    Institute of Scientific and Technical Information of China (English)

    胥全彬; A.Daugshies

    2001-01-01

    为区别从南京市江宁县采集的片形吸虫作典型形态虫体,应用随机扩增多态性DNA(RAPD)技术,对6株片形吸虫总DNA进行了扩增.结果,10条引物中有8条能产生扩增图谱,电泳图谱经聚类分析,与传统的分类结果一致,并表明来自江宁的片形吸虫既有形态典型的肝片形吸虫,也有形态不典型的大片形吸虫.

  14. A Preliminary Study on the Relationship Between the Indica-Japonica RAPD Differentiation of Parents and Heterosis in Dian Type Hybrid Rice

    Institute of Scientific and Technical Information of China (English)

    LONG Wen-hong; XU Ming-hui; ZHANG Shu-hua

    2002-01-01

    The relationship between the indica-japonica differentiation of parents and heterosis of hybridin rice was studied with 10 sterile lines, 10 maintainer lines of the sterile lines, 38 restorer lines of Dian typehybrid rice and 40 hybrids derived from the sterile lines and restorer lines. The results indicated that there wasa parabolic correlation between the indica-japonica differentiation of parents detected by RAPD markers andheterosis of hybrid rice. When certain indica-japonica differentiation of parents exists, the percentage of com-binations with positve heterosis and the mean heterosis of hybrids were high. But that didn't mean too muchdifference of parents could lead to stronger hetesosis, that meant overdiffernt parents seldom bring positve het-erotic hybrids. Suitable difference of percent of indica alleles of parents of stronger heterotic hybrid was 12 -16%.

  15. RAPD, SCAR and conserved 18S rDNA markers for a red-listed and endemic medicinal plant species, Knema andamanica (Myristicaceae).

    Science.gov (United States)

    Sheeja, T E; Anju, P R; Shalini, R S; Siju, S; Dhanya, K; Krishnamoorthy, B

    2013-04-01

    Knema andamanica is a red-listed endemic medicinal species of Myristicaceae restricted to Andaman and Nicobar (A&N) Islands, India. This species is used in tribal medicines and has immense bioprospective potential. With a view to generate suitable genomic markers for classification and identification, we have generated RAPD, SCAR and conserved 18S rDNA markers from K. andamanica. A unique 585 bp fragment, that distinguished it from seven other related species of Myristicaceae was first amplified using the random primer OPE 06 and converted to SCAR marker (GenBank accession # JN228256). The conserved sequences of 18S rDNA loci from K. andamanica were also amplified and sequenced (GenBank accession #JN228265). The sequence revealed deviations including 18 variable regions and 15 indels that were unique to K. andamanica. These markers can help in definite identification of K. andamanica even at the juvenile stages.

  16. Number of Drones Mated with Queen Determined Using RAPD Analysis%应用RAPD分析测定蜂王与雄蜂交配的数量

    Institute of Scientific and Technical Information of China (English)

    曾志将; 颜伟玉; 薛运波; 丁能水

    2003-01-01

    以卡尼鄂拉蜂(Apis mellifera carnica)和高加索蜂(Apis mellifera caucasica)的雄蜂精液对意大利蜜蜂(Apis mellifera ligustica)的处女蜂王进行人工授精,并利用11个引物进行随机扩增多态DNA(RAPD)标记分析.根据RAPD标记计算工蜂个体间的平均遗传相似系数,然后计算蜂王与雄蜂交配的数量.结果表明,RAPD标记是测定蜂王与雄蜂交配数量的一种有效方法.

  17. Identification of Lycopersicon spp. hybrids on the basis of morphological and molecular (RAPD properties as well as evaluation of resistance to Tomato Spotted Wilt Virus (TSWV

    Directory of Open Access Journals (Sweden)

    Zbigniew Rusinowski

    2014-01-01

    Full Text Available The aim of the study was to evaluate the plants obtained as a result of cultivated tomato crosses with wild species meant to transfer resistance to TSWV. Six viable plants were obtained from L. esculentum x L. chilense and L. esculentum x L. peruvianum crosses after the application of in vitro embryo culture. In terms of such morphological traits as growth habit of plants, size and shape of leaves, the length and colour of internodes in branching stems, the plants displayed intermediate traits, resembling, nonetheless, the wild form. RAPD analysis with 8 primers revealed that all the hybrids had bands typical of the paternal forms. This confirms the paternal component in hybrid development. As far as the resistance to Polish TSWV isolates is concerned, two hybrids exhibited a high level of resistance, similar to negative control, three hybrids - enhanced resistance and one hybrid was susceptible to TSWV infection.

  18. Caracterização de genótipos de cebola com a utilização de marcadores moleculares RAPD

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    Gerson Henrique Wamser

    Full Text Available A divergência genética foi avaliada entre quinze genótipos de cebola cultivados em Santa Catarina, com a utilização de marcadores moleculares RAPD. Onze oligonucleotídeos iniciadores da série Operon Technologies foram utilizados e produziram 35 marcadores, destes, 28 foram polimórficos. Os produtos da amplificação foram visualizados em gel de agarose 1,4%, corado com brometo de etídeo. Uma matriz de similaridade utilizando-se o coeficiente de Jaccard foi construída a partir dos dados moleculares. Um dendrograma foi gerado para melhor visualização da similaridade genética através do método de agrupamento UPGMA. Três grupos foram formados utilizando o coeficiente de similaridade 0,6 como ponto de corte. O primeiro grupo reuniu os genótipos Super Superprecoce e Gauchinha. O segundo grupo reuniu doze genótipos. Dentro desse grupo, os genótipos Bella Vista e Bella Dura foram os que apresentaram o maior coeficiente de similaridade, em torno de 0,89. Bela Vista e Superprecoce, Catarina e o híbrido Bella Vista, com coeficiente de similaridade de 0,88 entre os pares. O terceiro grupo apresentou apenas o genótipo Crioula Roxa, que obteve o menor valor (0,31 para o coeficiente de similaridade. Tendo em vista os resultados obtidos, cruzamentos entre os genótipos do primeiro e segundo grupo e destes com o genótipo Crioula Roxa, podem ser melhores por apresentarem maior divergência entre si. A técnica de RAPD mostrou-se eficaz na caracterização molecular dos genótipos de cebola, evidenciando que existe variabilidade entre os genótipos estudados.

  19. In vitro assessment of DNA damage after short- and long-term exposure to benzo(a)pyrene using RAPD and the RTG-2 fish cell line.

    Science.gov (United States)

    Castaño, Argelia; Becerril, Concepción

    2004-08-18

    Genotoxins present in the aquatic environment are often associated with the decline or disappearance of many wild populations. The hazard assessment of chemicals requires sensitive and specific tests to study the genotoxic effects in order to establish the maximum allowable chemical concentrations prior to the release to the environment. We have previously shown that an established fish cell line (RTG-2) together with the random amplified polymorphic DNA (RAPD) technique, can be used to detect alterations in the DNA caused by direct acting genotoxins. The current study takes this a step further and examines in the same system the effect of a pro-mutagen benzo(a)pyrene (B(a)P) at different concentrations (0.05, 0.1, and 0.5 microg/ml) and at different exposure periods (1, 2, 3, 15, and 30 days). After comparing DNA fingerprints from control and exposed cells, both qualitative and quantitative analysis show an increase in the instability in the DNA fingerprint of exposed cells over a time- and concentration-dependent manner. At the higher concentration (0.5 microg/ml) three out the four primers showed altered bands after 1 day of exposure, while after 3 days all used primers showed an altered pattern. At the lower concentration of B(a)P (0.05 microg/ml) the appearance of new bands was observed with a 100% level of reproducibility after 30 days of exposure suggesting an inheritance of the altered DNA. We conclude that this in vitro system is useful to evaluate genotoxic effects, both after acute and chronic exposures and of direct and non-direct acting genotoxins. Cultured cells can be considered as genetically homogenous populations. Therefore, in vitro systems permits us to undertake mechanistic studies avoiding the interference of polymorphisms inherent in the in vivo systems. Furthermore, the RTG-2 fish cell line combined with a RAPD assay could be used in studies of hazard identification in risk assessment protocols of chemicals.

  20. Genetic Homogeneity Revealed Using SCoT, ISSR and RAPD Markers in Micropropagated Pittosporum eriocarpum Royle- An Endemic and Endangered Medicinal Plant

    Science.gov (United States)

    Thakur, Julie; Dwivedi, Mayank D.; Sourabh, Pragya; Uniyal, Prem L.; Pandey, Arun K.

    2016-01-01

    Pittosporum eriocarpum Royle, a medicinally important taxon, is endemic to Uttarakhand region of Himalaya. It has become endangered due to over-collection and the loss of habitats. As raising plants through seeds in this plant is problematic, a reliable protocol for micropropagation using nodal explants has been developed. High shoot regeneration (95%) occurred in MS medium augmented with BA 0.4mg/l in combination IBA 0.6mg/l. In vitro regenerated shoots were rooted in MS medium supplemented with three auxins, of which 0.6 mg/l indole butyric acid proved to be the best for rooting (90%) with maximum number of roots per shoot. Thereafter, rooted plants were hardened and nearly 73% of rooted shoots were successfully acclimatized and established in the field. Start codon targeted (SCoT), inter simple sequence repeats (ISSR) and random amplified polymorphic DNA (RAPD) markers were used to validate the genetic homogeneity amongst nine in vitro raised plantlets with mother plant. DNA fingerprints of in vitro regenerated plantlets displayed monomorphic bands similar to mother plant, indicating homogeneity among the micropropagated plants with donor mother plant. The similarity values were calculated based on SCoT, ISSR and RAPD profiles which ranged from 0.89 to 1.00, 0.91 to 1.00 and 0.95 to 1.00 respectively. The dendrograms generated through Unweighted Pair Group Method with arithmetic mean (UPGMA) analysis revealed 97% similarity amongst micropropagated plants with donor mother plant, thus confirming genetic homogeneity of micropropagated clones. This is the first report on micropropagation and genetic homogeneity assessment of P. eriocarpum. The protocol would be useful for the conservation and large scale production of P. eriocarpum to meet the demand for medicinal formulations and also for the re-introduction of in vitro grown plants in the suitable natural habitats to restore the populations. PMID:27434060

  1. Genetic Homogeneity Revealed Using SCoT, ISSR and RAPD Markers in Micropropagated Pittosporum eriocarpum Royle- An Endemic and Endangered Medicinal Plant.

    Science.gov (United States)

    Thakur, Julie; Dwivedi, Mayank D; Sourabh, Pragya; Uniyal, Prem L; Pandey, Arun K

    2016-01-01

    Pittosporum eriocarpum Royle, a medicinally important taxon, is endemic to Uttarakhand region of Himalaya. It has become endangered due to over-collection and the loss of habitats. As raising plants through seeds in this plant is problematic, a reliable protocol for micropropagation using nodal explants has been developed. High shoot regeneration (95%) occurred in MS medium augmented with BA 0.4mg/l in combination IBA 0.6mg/l. In vitro regenerated shoots were rooted in MS medium supplemented with three auxins, of which 0.6 mg/l indole butyric acid proved to be the best for rooting (90%) with maximum number of roots per shoot. Thereafter, rooted plants were hardened and nearly 73% of rooted shoots were successfully acclimatized and established in the field. Start codon targeted (SCoT), inter simple sequence repeats (ISSR) and random amplified polymorphic DNA (RAPD) markers were used to validate the genetic homogeneity amongst nine in vitro raised plantlets with mother plant. DNA fingerprints of in vitro regenerated plantlets displayed monomorphic bands similar to mother plant, indicating homogeneity among the micropropagated plants with donor mother plant. The similarity values were calculated based on SCoT, ISSR and RAPD profiles which ranged from 0.89 to 1.00, 0.91 to 1.00 and 0.95 to 1.00 respectively. The dendrograms generated through Unweighted Pair Group Method with arithmetic mean (UPGMA) analysis revealed 97% similarity amongst micropropagated plants with donor mother plant, thus confirming genetic homogeneity of micropropagated clones. This is the first report on micropropagation and genetic homogeneity assessment of P. eriocarpum. The protocol would be useful for the conservation and large scale production of P. eriocarpum to meet the demand for medicinal formulations and also for the re-introduction of in vitro grown plants in the suitable natural habitats to restore the populations.

  2. 大麻RAPD研究的方法学探讨%An Effective Procedure for the RAPD Analysis of Hemp Cannabis sativa L.

    Institute of Scientific and Technical Information of China (English)

    宋书娟; Robert. C.Clarke; 邵宏

    2001-01-01

    中国有丰富的大麻资源.目前,在我国对于大麻的研究多集中在形态和化学成分等方面,而分子遗传水平的研究至今尚未见报道.RAPD分子标记技术是适合于大麻分子水平研究的首选方法.本文以采集多年的大麻植物干标本的叶为研究材料,分别对Mg2+,dNTPs,Taq DNA酶的浓度,和退火温度和退火时间以及反应循环数等条件进行了检测,得到了适合大麻RAPD研究的理想条件.实验结果表明,用干标本来进行一定的分子遗传研究是可行的,且结果也相对稳定.%China has a great resource of Cannabis. Research on the taxonomy and morphology of Chinese Cannabis has been carried out, but so far no molecular genetic research has been published. Random amplified polymorphic DNA (RAPD) is a suitable technique for molecular genetic research on Cannabis. In this experiment,using Cannabis herbarium specimens as a source of genetic materials, the correlative conditions of the Polymerase chain reaction (PCR), (i.e., gradient density of Mg2+, dNTPs, Taq DNA polymerase, anneal temperature, anneal time and reaction cycles) were examined separately. An effective procedure for the RAPD analysis of Cannabis was obtained.

  3. A Comparison of the Effectiveness of Sodium Stibogluconate Monotherapy to Sodium Stibogluconate and Paromomycin Combination for the Treatment of Severe Post Kala Azar Dermal Leishmaniasis in South Sudan – A Retrospective Cohort Study

    Science.gov (United States)

    Abongomera, Charles; Gatluak, Francis; Buyze, Jozefine; Ritmeijer, Koert

    2016-01-01

    Background Post-kala-azar dermal leishmaniasis (PKDL) is a common dermatological complication following successful treatment of Visceral Leishmaniasis (VL) caused by Leishmania donovani. PKDL presents as macular, papular, nodular or mixed skin rash on sun-exposed body parts. Patients are not ill unless there are complications due to mucosal involvement or ulceration. As PKDL in East Africa is typically self-healing, and treatment is long and with significant adverse events, only severe and complicated cases are treated. Studies to determine optimal treatment of PKDL are rare and based on small cohorts. Since 1989, Médecins Sans Frontières is treating severe PKDL within VL treatment programmes in South Sudan. Treatment was initially with sodium stibogluconate (SSG) monotherapy and since 2002 with a combination of SSG and paromomycin (PM). SSG monotherapy (20 mg/kg/day for a minimum of 30 days) was provided in primary health units, and the combination of PM (15 mg sulphate/kg/day for 17 days) plus SSG (30 mg/kg/day for a minimum of 17 days) was provided in secondary health facilities. Methodology/Principal Findings By retrospective analysis of routinely collected programme data we compared the effectiveness (outcome and treatment duration) of both regimens. Between 2002 and 2008, 422 patients with severe PKDL were treated; 343 received SSG and 79 SSG/PM combination. The cure rate was significantly better with combination treatment when compared to monotherapy (97% vs. 90%; odds ratio [OR], 7.6; p = 0.02), treatment duration was shorter (mean 34 days vs. 42 days; p = 0.005), and defaulter rate was lower (3% vs. 9%; OR, 0.3; p = 0.03). There was no significant difference in death rate (0% vs. 1%; p = 0.5). Conclusions/Significance We found that SSG/PM combination therapy resulted in more favourable outcomes than SSG monotherapy. An additional advantage is the lower cost of the combination therapy, due to the shorter treatment duration. A combination of SSG and PM is

  4. Assessment of genetic diversity on a sample of cocoa accessions resistant to witches' broom disease based on RAPD and pedigree data Avaliação da diversidade genética em uma amostra de acessos de cacau resistentes à doença vassoura-de-bruxa, com base em dados de RAPD e pedigree

    Directory of Open Access Journals (Sweden)

    Ronaldo Carvalho dos Santos

    2005-01-01

    Full Text Available Genetic diversity in cocoa (Theobroma cacao L. has been assessed based on morphological and molecular markers for germplasm management and breeding purposes. Pedigree data is available in cocoa but it has not been used for assessing genetic relatedness. The geneitic diversity of 30 clonal cocoa accessions resistant to witche´ broom disease, from the CEPEC series, were studied on the basis of RAPD data and pedigree information. Twenty of these accessions descend from the TSA-644 clone, originated from a cross between the Upper Amazon germplasm called Scavina-6, the main source of resistance to witches' broom disease, and IMC-67. The ten remaining clones come from different sources including Amazon and Trinitario germplasm. RAPD data was collected using 16 primers and pedigree information was obtained from the International Cocoa Germplasm Database. Genetic similarities, genetic distances and coefficient of parentage were calculated using available software. Relatively low genetic diversity was observed in this germplasm set, probably because of great genetic relatedness amongst accessions studied and the poor representation of the germplasm. The TSA-644 descendants were more diverse than the other accessions used in the study. This might be due to the origin of the TSA clone, which was derived from highly divergent genotypes. Association between genetic similarities based on RAPD data and coefficient of parentage, based on pedigree data, was very low, probably due to the homogeneity of the breeding stocks and poor pedigree information. These findings are useful to cocoa breeders in planning crosses for the development of hybrid and clonal cultivars.A diversidade genética em cacau (Theobroma cacao L., embasada em dados morfológicos e em marcadores moleculares, tem sido avaliada com fins de manejo de germoplasma e uso no melhoramento genético. Dados de genealogia de cacau, embora disponíveis, não têm sido utilizados. Foi analisada a

  5. Genetic similarity among Agistemus pallinii Matioli et al (Acari: Stigmaeidae) found in citrus orchards in Vicosa, Minas Gerais state, Brazil

    Energy Technology Data Exchange (ETDEWEB)

    Matioli, Andre L. [Instituto Biologico, Campinas, SP (Brazil). Lab. de Entomologia Economica], e-mail: almatioli@biologico.sp.gov.br; Pallini, Angelo [Universidade Federal de Vicosa (UFV), MG (Brazil). Dept. de Biologia Animal], e-mail: pallini@ufv.br; Tavares, Mara G. [Universidade Federal de Vicosa (UFV), MG (Brazil). Dept. de Biologia Geral], e-mail: mtavares@ufv.br

    2009-03-15

    Stigmaeidae are very important predators of mite and insect pests on several crops in Brazil. It is considered the second most important family of predatory mites in citrus orchards in Brazil. However, their identification, especially that of the members of the genus Agistemus, is rather difficult based only on morphology. Hence, this study describes the use of random amplified polymorphic DNA (RAPD-PCR) markers to determine the genetic similarity of an Agistemus pallinii Matioli et al population found in 2004 in a citrus orchard in Vicosa, Minas Gerais State, Brazil, preying on Panonychus citri (McGregor). Amplifications were performed with 12 random primers (OPAA8, OPAA19, OPAB1, OPAB5, OPAB18, OPAC9, OPAC17, OPAC19, OPAD10, OPAE9, OPAE12 and OPAE17), which generated 119 bands, with 53.8% polymorphism. The coefficients of genetic similarity among the individuals ranged from 0.68 to 0.99, indicating a high genetic similarity among them. The 3D projection analysis clustered the majority of individuals confirming their high similarity. Though individuals of A. pallinii are minute ({+-} 360 {mu}m long), the PCR-RAPD technique can still be used for their identification, complementing morphological analyses or for comparison of populations collected in different geographic regions. This is the fi rst molecular study carried out with stigmaeid mites. (author)

  6. Abu al-Layth al-Libi

    Science.gov (United States)

    2015-02-01

    He has described at length the various methods to which the prisoners were subjected, including sleep depravation, nudity and the use of electrodes...suspects of the Riyadh bombing acknowledge their involvement on television , he was under the impression that they too had been subjugated to this... sleeping , al-Libi, al-Zawi and `Abd al-Karim managed to escape by jumping from the six-meter-tall wall of al-Ruways.38 Their breakout led to a

  7. RAPD em Pau-rosa (Aniba rosaeodora Ducke: adaptação do método para coleta de amostras in situ, ajuste das condições de PCR e apresentação de um processo para selecionar bandas reprodutíveis RAPD in Rosewood (Aniba rosaeodora Ducke: adequation of a method for in situ sample collecting, PCR adjustments and presentation of a procedure to select reproducible amplified fragments

    Directory of Open Access Journals (Sweden)

    Ronaldo Pereira Santos

    2007-06-01

    Full Text Available O Pau-rosa (Aniba rosaeodora Ducke é uma espécie importante economicamente para a Região Amazônica, porque sua madeira é fonte de linalol, insumo utilizado pelas perfumarias. Esta espécie foi explorada durante décadas e, ainda assim, o conhecimento acerca da diversidade genética intra-específica é muito restrito. Foram objetivos deste trabalho: 1 validar um protocolo para coleta de folhas de pau-rosa que permitisse preservar a integridade do DNA até a estocagem em "freezer"; 2 selecionar um protocolo para extração de DNA em quantidade e qualidade adequadas para geração de bandas RAPD e 3 desenvolver um critério para avaliar o grau de reprodutibilidade que pudesse auxiliar a seleção de bandas RAPD úteis para análises de diversidade genética. Imediatamente após a coleta, as folhas foram acondicionadas em tubos de polietileno com sílica gel e aí permaneceram por até 10 dias. Foram testados três protocolos para a extração de ácidos nucléicos destas folhas, condições ideais para as PCR e a reprodutibilidade dos padrões RAPD. Critérios para a eliminação das bandas que mais contribuíram para o afastamento dos resultados do ideal da reprodutibilidade total foram desenvolvidos e a significância estatística das diferenças geradas pela aplicação dos critérios ao conjunto de dados foi testada. DNA com qualidade e em quantidade suficiente para a geração de padrões RAPD, nas condições ideais definidas para as PCRs, foi obtido. A eliminação de bandas com reprodutibilidade menor que 70% não diferiu do controle. A eliminação de bandas com reprodutibilidade menor que 90% diferiu dos demais tratamentos em todos os arranjos testados (P Rosewood (Aniba rosaeodora Ducke is one of the economically valuable species in the Amazon region, because it is the principal source of linalool which is demanded by the perfumery industry. This species was submitted to hard exploitation along the past decades and besides this

  8. 辣椒RAPD系统的建立及在杂种纯度鉴定中的应用%Establishment of an Efficient RAPD Protocol in Pepper and Its Application in Gen etic Purity Testing of F1 Seeds

    Institute of Scientific and Technical Information of China (English)

    黄三文; 张宝玺; 等

    2001-01-01

    n this report,a simple and efficient protocol for RAPD assay in pepper was est ablished through optimization in three main procedures-DNA extraction,PCR,el ectrophoresis.The protocol was employed in screening RAPD markers for genetic pu rity testing of‘Zhongjiao’serial hybrid cultivars.A total of 12 stable and str ong RAPDs were identified to distinguish all 9 F1 s from their parental lines.%对辣椒RAPD反应的各个环节, 包括DNA提取、PCR反应和电泳检 测进行了筛选和优化,确定了一个简单、高效、相对稳定的RAPD系统,并筛选出12个较稳定 的RAPD标记。可以用于“中椒”系列辣椒的9个杂交种的纯度鉴定。

  9. "Use of Random Amplified Polymorphic DNA Polymerase Chain Reaction (RAPD-PCR and ITS2 PCR assays for differentiation of populations and putative sibling species of Anopheles fluviatilis (Diptera: Culicidae in Iran"

    Directory of Open Access Journals (Sweden)

    SR Naddaf Dezfouli

    2002-09-01

    Full Text Available Anopheles fluviatilis complex is known to be a vector of malaria in Iran. Since mosquitoes of this species cover a wide geographical range in Iran, they might have evolved into different separated populations. Random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR assay was used to differentiate geographic populations of this species. DNA was extracted from individual mosquitoes from 8 localities in 4 south and southeast provinces and amplified in PCR reactions using 18 single primers of arbitrary nucleotide sequence. Results of RAPD-PCR showed that Kazeroun populations could simply be differentiated from other populations using a diagnostic fragment amplified with primer UBC-306. But other populations could not be differentiated either visually or by means of statistical analysis. Moreover ITS2 fragments of some selected specimens were amplified using a pair of universal primer and sequenced as a key standard for detection of putative sibling species. Sequence analysis of the ITS2 fragments revealed a very high (100% homology among the populations. These findings are crucial in epidemiological studies concerning relatedness of geographic populations and vector movement in the region. Results of RAPD-PCR and ITS2 analysis suggest that this taxon in Iran comprises of only one species with a low genetic variation among geographic populations.

  10. [Genetic diversity and subdivision parameters of Colias crocea Fourc. and C. erate Esp. (Lepidoptera, Pieridae) in Crimea according to allozyme and RAPD-PCR analyses].

    Science.gov (United States)

    Milovanov, A E; Simchuk, A P

    2008-01-01

    Some parameters of genetic diversity and subdivision were tested in syntopic Crimean populations of two species of the sulfur butterllies, Colias crocea Fourc. and C. erate Esp. by allozyme and RAPD-PCR analyses. Genetic diversity in each of the species compared appeared to be approximately equal: the portion of polimorphic loci P95 (99) is 67% for C. crocea and is 61% for C. erate. Paratypic forms of both species do not differ from nominative ones by parameters of genetic diversity. The mean expected heterozygosities (H'(e)) in the C. crocea, C. erate and their paratypic forms by the RAPD-PCD data are 24.3 +/- 4.4%, 21.7 +/- 4.2%, 26.4 +/- 4.1%, respectively. The observed heterozygosities by the allozyme data in other samples are considerably less than theoretically expected ones (H(o) = 15.6 +/- 3.3%, H(e) = 50.5 +/- 4.5% for C. crocea; H(o) = 17.95 +/- 6.15%, H(e) = 50.8 +/- 8% for C. erate, and H(o) = 24.2 +/- 7.5%, H(e) = 50.9 +/- 8.7% for paratypic forms, respectively). The mean observed heterozygosity in the paratypic forms of C. erate (f. androconiata, f. chrysodona, f. edusoides) is almost two times higher than that of the f. chlorodona and f. eratoides. Paratypic forms of both species differ from nominative ones (particularly from that of C. crocea) by the level of intrapopulation inbreeding and by the degree of gene flow (F = 0.691 for C. crocea, 0.646 for C. erate, and 0.524 for paratypic forms). One can suppose a significant gene exchange among adjacent populations of C. crocea and C. erate (Wright's F(ST) = 0.155 for 6 allozyme and 18 RAPD loci, Nm = 1.363). Nei's similarity coefficients (S) varies from 0.7554 among nominative C. crocea and C. erate (D(N) = 0.28) to 0.8092 among C. crocea and paratypic forms (D = 0.21) and to 0.8936 among C. erate and paratypic forms of both species (D(N) = 0.11). Paratypic forms with rounded valve margin preliminarily identified as C. crocea revealed a considerable degree of similarity to the paratypic forms of C

  11. Analysis of the genetic polymorphism of Paracoccidioides brasiliensis and Paracoccidioides cerebriformis "Moore" by random amplified polymorphic DNA (RAPD and 28S ribosomal DNA sequencing: Paracoccidioides cerebriformis revisited Análise do polimorfismo genético do Paracoccidioides brasiliensis e Paracoccidioides cerebriformis "Moore" pela técnica de amplificação aleatória do polimorfismo do DNA (RAPD e sequenciamento do DNA ribossomal 28S: Paracoccidioides cerebriformis revisitado

    Directory of Open Access Journals (Sweden)

    Sarah Desirée Barbosa Cavalcanti

    2005-06-01

    Full Text Available Our purpose was to compare the genetic polymorphism of six samples of P. brasiliensis (113, 339, BAT, T1F1, T3B6, T5LN1, with four samples of P. cerebriformis (735, 741, 750, 361 from the Mycological Laboratory of the Instituto de Medicina Tropical de São Paulo, using Random Amplified Polymorphic DNA Analysis (RAPD. RAPD profiles clearly segregated P. brasiliensis and P. cerebriformis isolates. However, the variation on band patterns among P. cerebriformis isolates was high. Sequencing of the 28S rDNA gene showed nucleotide conservancy among P. cerebriformis isolates, providing basis for taxonomical grouping, and disclosing high divergence to P. brasiliensis supporting that they are in fact two distinct species. Moreover, DNA sequence suggests that P. cerebriformis belongs in fact to the Aspergillus genus.Nosso propósito foi comparar o polimorfismo genético de seis amostras de P. brasiliensis (113, 339, BAT, T1F1, T3B6, T5LN1, com quatro amostras de P. cerebriformis (735, 741, 750, 361 do laboratório de micologia do Instituto de Medicina Tropical de São Paulo, utilizando a técnica de Amplificação Aleatória do Polimorfismo de DNA (RAPD. O perfil de bandas do RAPD diferenciou claramente os isolados de P. brasiliensis de P. cerebriformis. Entretanto, ocorreu uma variação significativa no padrão de bandas das amostras de P. cerebriformis. O sequenciamento do gene ribossomal 28S revelou seqüências de nucleotídeos bastante conservadas entre os isolados de P. cerebriformis, fornecendo subsídio para o agrupamento taxonômico destas amostras, diferenciando estas de P. brasiliensis e mostrando que de fato são espécies distintas. A seqüência de DNA sugere que P. cerebriformis pertence ao gênero Aspergillus.

  12. Identification of Seed Purity of Oilseed Sunflower (Helianthus annuus L.) Hybrid A15 with RAPD%杂交油葵A15种子纯度的RAPD鉴定

    Institute of Scientific and Technical Information of China (English)

    莫结胜; 刘杰; 刘公社

    2001-01-01

    从杂交油葵A15及其亲本中提取基因组DNA,用180个RAPD随机引物进行扩增,从中筛选出3个可将亲本和子代区分开的引物OPD09、OPD12和OPK12。OPD09产生亲本互补的特征带OPD09-1470bp、OPD09-870bp;OPD12产生母本特征带OPD12-1230bp,OPK12产生父本特征带OPK12-1540bp、OPK12-940bp,上述谱带均在子代中出现。以单引物(OPD09)和双引物(OPD12和OPK12)产生的这两组特征谱带作为分子标记分别对杂交油葵种子纯度进行鉴定得到了一致的结果,并与大田纯度检测结果相一致。用RAPD鉴定杂交油葵种子纯度是可行的。%The genomic DNAs extracted from young leaves of oilseed sunflowerhybrid A15 and its parents were analyzed using polymerase chain reaction (PCR) with arbitrary 10-mer primers to generate random amplified polymorphic DNA (RAPD) markers. Among 180 RAPD primers, three primers, OPD09, OPD12, OPK12, which could generate differential RAPD profiles among A15 and its parents, were screened. Primer OPD09 could generate parental complementary characteristic bands: OPD09-1470bp, OPD09-870bp; primer OPD12 could generate maternal characteristic band OPD12-1230bp; primer OPK12 could generate paternal characteristic bands OPK12-1540bp and OPK12-940bp. All these characteristic bands screened were found in A15 hybrids. These bands can be used as RAPD markers to identify seed purity of A15. It is viable to identify oilseed sunflower hybrids seed purity with RAPD.

  13. 用随机扩增多态DNA(RAPD)技术研究不同地理株埃及伊蚊的分化%Differentiation of Aedes aegypti with Genetic Polymorphisms Detected By the Random Amplified Polymorphism DNA Polymerase Chain Reaction (RAPD-PCR)

    Institute of Scientific and Technical Information of China (English)

    周光智; 赵彤言; 李春晓; 朱礼华; 薛健

    2003-01-01

    目的用RAPD技术对实验室饲养的广东、海南、台湾和印尼Baro等4个不同地理株的8只雌蚊进行随机扩增多态DNA分析.方法随机扩增多态性DNA(RAPD)技术.结果选用20个随机引物进行扩增,有8个引物表现清晰的RAPD谱带并呈显著多态性.UPGMA法构建的分子系统树表明埃及伊蚊4个地理株之间存在着一定程度的遗传分化.结论用RAPD方法可以区分不同地理株埃及伊蚊.

  14. Factores de riesgo asociados al embarazo ectópico

    Directory of Open Access Journals (Sweden)

    Jónathan Hernández Núñez

    2013-12-01

    Full Text Available Fundamento: el embarazo ectópico se ha duplicado en la mayor parte de los países industrializados y representa una de las principales causas de muerte materna. Objetivo: identificar factores de riesgo que predisponen al embarazo ectópico.Métodos: estudio de casos y controles realizado en un universo de 132 pacientes del Municipio Habana del Este, operadas de embarazo ectópico en el Hospital Dr. Luís Díaz Soto desde el 2009 al 2011. Se escogió una muestra al azar de 66 pacientes que constituyó el grupo estudio; se contó con otro grupo control de igual cantidad de pacientes no operadas de embarazo ectópico en dicho período. Se analizaron variables relacionadas con factores de riesgo sociodemográficos, ginecobstétricos y quirúrgicos, conducta sexual y hábito de fumar. Se empleó la media, desviación estándar, porcentaje, test de Student, estadígrafo chi cuadrado y odds ratio. Resultados: el embarazo ectópico fue más frecuente en mujeres de color de piel blanca, entre 25 y 34 años de edad; el antecedente de enfermedad inflamatoria pélvica, endometriosis, tres o más abortos provocados, así como la historia de infertilidad, embarazo ectópico previo, inicio precoz de las relaciones sexuales, múltiples compañeros sexuales y el empleo de dispositivos intrauterinos fueron factores predisponentes para el embarazo ectópico; no así la escolaridad, ocupación, paridad, apendicectomía y cirugía tubárica previa, ni tabaquismo.Conclusiones: existen múltiples factores de riesgo que predisponen al embarazo ectópico y que contribuyen al incremento de la morbimortalidad materna, muchos de los cuales son susceptibles de ser modificados desde la atención primaria de salud.

  15. Characterization of fusion products from protoplasts of yeasts and their segregants by electrophoretic karyotyping and RAPD Caracterização de produtos de fusão de protoplastos de leveduras e seus segregantes via cariotipagem eletrodorética e RAPD

    Directory of Open Access Journals (Sweden)

    Cleide Viviane Buzanello Martins

    1999-01-01

    Full Text Available In order to characterize fusion products from yeast protoplasts and their segregants, with important features to the wine making industry, electrophoretic karyotyping and RAPD (Random Amplified Polymorphic DNA were utilized. Electrophoretic karyotyping was performed by the CHEF ("contour-clamped homogeneous electric field electrophoresis" method, which allowed the detection of chromosomal band complementation in fusion products and the presence of patterns of both parental and intermediary strains in segregants. By utilizing two primers, an amplification pattern of DNA fragments was obtained. While fusion products (diploid showed a pattern of complementary bands, segregants showed bands of either parental strains or even intermediary bandsCom o objetivo de caracterizar os produtos de fusão de protoplastos de leveduras com características de importância para a indústria vinícola e seus segregantes, foram empregadas as técnicas de separação de bandas cromossômicas por eletroforese e de RAPD (amplificação ao acaso de DNA polimórfico. O cariótipo eletroforético foi realizado pelo método CHEF ("contour-clamped homogeneous eletric field eletrophoresis", constatando-se a complementação de bandas cromossômicas no produto de fusão e padrões de ambos os parentais e padrões intermediários nos segregantes. A análise do padrão de amplificação dos fragmentos de DNA com dois primers evidenciou um padrão de bandas complementares nos produtos de fusão (diplóide e padrão de bandas de um e de outro parental ou mesmo bandas intermediárias nos segregantes.

  16. Analysis the Genetic Divergence of Botrytis by RAPD%葡萄孢菌全基因组DNA遗传多态性RAPD分析

    Institute of Scientific and Technical Information of China (English)

    吴德喜; 王扬; 王学英; 张立新; 黄国品; 孔令平

    2000-01-01

    @@ 应用随机扩增多态性DNA(RAPD)分子标记技术,从Operon Technologies公司的OPA和OPB组随机引物中筛选出OPA01,OPA02,OPA05,OPA06,OPA07,OPA11,OPA12,OPA13,OPA18,OPA19,OPB01,OPB06,OPB20等13个寡聚核苷酸引物,对采自云南、西藏、四川等地的B01,B02两种不同寄主的B.cinerea和B03(B.vieiao),B05(B.convolvta),B06(B.linzhii),B10(B.paeoniae),B11(B.paeonialla),B12(B.yunnanensis),B13(B.multiformis),B14(B.acladiopsis)等10个Botrytis属菌种以及B09(Homobotrys fagapyruma)全基因组DNA进行随机扩增,进行遗传相似性分析,现将结果简要报告如下:

  17. Prevalence of Listeria species in camel sausages from retail markets in Aydin province in Turkey and RAPD analysis of Listeria monocytogenes Isolates

    Directory of Open Access Journals (Sweden)

    Ozbey Gokben

    2006-06-01

    Full Text Available Abstract Samples were taken from 100 camel sausages from the different retail markets in Aydin province in the south-west of Turkey and they were tested for the presence of Listeria spp by biochemical methods. Samples were enriched using Listeria Enrichment Broth and they were inoculated onto Listeria Selective Agar. Listeria monocytogenes was isolated from nine samples (9%, Listeria innocua from 14 samples (14% and Listeria welshimeri from two samples(2%. A 701 bp fragment of listeriolysin O sequence for L. monocytogenes was amplified using specific primers by polymerase chain reaction (PCR for confirmation of the identification. A random primer (OPA-11 was used in a random amplified polymorphic DNA (RAPD assay. This detected five different band profiles amongst the L. monocytogenes isolates, indicating a relatively large amount of genetic heterogeneity amongst the nine isolates. The study has highlighted the need for improved strategies for food safety, in particular appropriate hygienic precautions to avoid contamination of sausage during the manufacturing process and appropriate preservation techniques during storage and transport, to prevent transmission of Listeria spp to consumers at home and abroad.

  18. 卡瓦胡椒及胡椒的RAPD聚类分析%Study on the Clustering Analysis of Piper methysticum and Piper nigrum by RAPD

    Institute of Scientific and Technical Information of China (English)

    施江; 辛莉; 郑楷; 李建国; 郑学勤

    2005-01-01

    目的:通过RAPD分析搞清楚卡瓦胡椒与胡椒及胡椒属其它近缘野生种的亲缘关系.方法和结果:采用随机引物80条进行筛选,用从80条随机引物中入选的20条引物,对卡瓦胡椒和胡椒属共计28份材料进行RAPD扩增,均能产生清晰的扩增谱带.重复1~2次,结果稳定可靠.20个引物共扩增出170个条带,其中多态性条带有20条,占总扩增条带数的12%.RAPD分析结果显示在相似系数0.36处对28份种质可划分为6个类型,其中卡瓦胡椒被单独聚为一类,说明卡瓦胡椒与胡椒及其它近缘野生种的亲缘关系有一定的距离.

  19. Genetic differences between wild and hatchery populations of Diplodus sargus and D. vulgaris inferred from RAPD markers: implications for production and restocking programs design.

    Science.gov (United States)

    Pereira, J C; Lino, P G; Leitão, A; Joaquim, S; Chaves, R; Pousão-Ferreira, P; Guedes-Pinto, H; dos Santos, M Neves

    2010-01-01

    Restocking and stock enhancement programs are now recognized as an important tool for the management of fishery resources. It is important, however, to have an adequate knowledge on the genetic population structure of both the released stock and the wild population before carrying out such programs. In this study, random amplified polymorphic DNA (RAPD) markers were applied to assess genetic diversity and population structure of wild and hatchery populations of the white seabream Diplodus sargus and the common two-banded seabream D. vulgaris (Sparidae). The estimated values for intrapopulation genetic variation, measured using the percentage of polymorphic loci (%P), Shannon index (H'), and Nei's gene diversity (h), showed high values for all populations. The percentage of genetic variation within D. sargus and D. vulgaris populations, based on coefficient of gene differentiation, reached 82.5% and 90% of the total genetic variation, respectively. An undeniable decrease in genetic variation was found in both hatchery populations, particularly in D. sargus, compared to the wild ones. However, the high values of variation within all populations and the low levels of genetic variation among populations did not indicate inbreeding or depression effects, thus indicating a fairly proper hatchery management. Nevertheless, the results of this study highlight the importance of monitoring the genetic variation of hatchery populations, particularly those to be used in restocking programs. The creation of a genetic baseline database will contribute to a more efficient conservation management and to the design of genetically sustainable restocking programs.

  20. RAPD Analysis of Seven Vespertilionid Bats (Chiroptera: Vespertilionidae) in Henan Province%河南省蝙蝠科7种蝙蝠的RAPD分析

    Institute of Scientific and Technical Information of China (English)

    余燕; 马金友; 王艳梅; 牛红星

    2007-01-01

    为了解蝙蝠科种间亲缘关系,采用随机引物对河南省蝙蝠科7种蝙蝠进行DNA多态性研究,从20个随机引物中优化出12个引物对基因组DNA进行扩增,共扩增出223条DNA谱带,平均每个引物扩增出18.6条谱带.RAPD聚类结果表明,种间亲缘关系较远,种内亲缘关系较近.对同种蝙蝠而言,同一地理区域的蝙蝠个体之间分化较小,不同地理区域的蝙蝠个体之间分化较大.同时对鼠耳蝠属和长翼蝠亚科的分类地位也进行了讨论.

  1. 不同地理区域蛤蚧的RAPD分析%RAPD Analysis on Genetic Divergence and Phylogenesis of Gekko gecko from Different Areas

    Institute of Scientific and Technical Information of China (English)

    秦新民; 梁燕妮; 黄夕洋; 庞广福

    2005-01-01

    应用随机扩增多态性DNA(RAPD)技术,对6个不同地域(河池、南宁、桂林、百色、越南、泰国)的蛤蚧(Gekko gecko)进行了遗传多样性和系统发育分析.21个RAPD引物共扩增了218个位点,片段大小在200~2 000 bp,其中184个是多态位点,占84.4%.地域之间的遗传距离指数在0.011 2~0.963 1,遗传相似性系数在0.381 7~0.988 8之间,根据遗传距离指数和遗传相似性系数,用NTSYSpc 2.10软件包中的UPGMA法构建了系统聚类图,结果均显示南宁地区、桂林地区、百色地区和河池地区先聚在一起,再和越南聚在一起,最后和泰国群体聚类.这与形态地理分布特征相一致.

  2. 大花蕙兰基因组DNA提取及RAPD反应条件探索%Genomic DNA Extraction and RAPD Protocols for Cymbidium hybridium

    Institute of Scientific and Technical Information of China (English)

    李冬梅; 朱根发; 叶庆生

    2006-01-01

    用SDS、CTAB和改良CTAB法分别提取大花蕙兰叶片基因组DNA,发现以改良CTAB法提取的DNA纯度高,产率也较高.以5'-GGTGCTCCGT-3'(BA440)为随机引物,并以大花蕙兰品种‘金杯'(Cymbidium hybridium cv.Hiroshima Golden Cup"Sunny Moon")的DNA为模板,对RAPD(Random Amplified Polymorphic DNA)反应体系进行了优化研究,结果表明,25μl反应体系中,Mg2+、Taq DNA聚合酶、引物、模板DNA和dNTP 5种主要成分的适宜浓度或用量分别是:2.0 mmol/L、1.0 U、0.20μmol/L、25 ng和0.20 mmol/L.扩增程序优化为:94℃预变性4 min;94℃变性0.5 min,37℃退火1 min,72℃延伸1 min,40个循环;最后72℃延伸7 min.

  3. 大麻性别的RAPD和SCAR分子标记%RAPD and SCAR Molecular Markers of Sexuality in the Dioecious Species Cannabis sativa L.

    Institute of Scientific and Technical Information of China (English)

    陈其军; 韩玉珍; 傅永福; 赵德刚; 国凤利; 孟繁静; 刘卫平

    2001-01-01

    利用随机扩增多态性DNA (random amplified polymorphic DNA, RAPD)技术获得与大麻性别连锁的分子标记.将10株雄性大麻或10株雌性大麻的单个DNA样品等量混合分别组成雄性或雌性DNA池(DNA pool),以提供具有相同遗传背景的雌、雄性DNA样品.每个随机引物分别用三个不同的循环程序进行PCR扩增.在30个随机引物中,用引物401扩增得到一条约2.5kb雄性多态性片段.对该片段进行了克隆和序列分析,并根据序列分析结果将上述RAPD分子标记转化为重复性和特异性更好的SCAR(sequence characterized amplified regions)分子标记.

  4. Characteristics of PCR-SSCP and RAPD-HPCE methods for identifying authentication of Penis et testis cervi in Traditional Chinese Medicine based on cytochrome b gene.

    Science.gov (United States)

    Li, Mingcheng; Gao, Lijun; Qu, Li; Sun, Jingyu; Yuan, Guangxin; Xia, Wei; Niu, Jiamu; Fu, Guilian; Zhang, Lihua

    2016-07-01

    The use of Penis et testis cervi, as a kind of precious Traditional Chinese Medicine (TCM), which is derived from dry deer's testis and penis, has been recorded for many years in China. There are abundant species of deer in China, the Penis et testis from species of Cervus Nippon and Cervus elaphusL were authentic, others species were defined as adulterant (different subspecies of deer) or counterfeits (different species). Identification of their origins or authenticity becomes a key in controlling the herbal products. A modified column chromatography was used to extract mitochondrial DNA of dried deer's testis and penis from sika deer (C. Nippon) and red deer (C. elaphusL) in addition to adulterants and counterfeits. Column chromatography requires for a short time to extract mitochondrial DNA of high purity with little damage of DNA molecules, which provides the primary structure of guarantee for the specific PCR; PCR-SSCP method showed a clear intra-specific difference among patterns of single-chain fragments, and completely differentiate Penis et testis origins from C. Nippon and C. elaphusL. RAPD-HPCE was based on the standard electropherograms to compute a control spectrum curve as similarity reference (R) among different samples. The similarity analysis indicated that there were significant inter-species differences among Penis et testis' adulterant or counterfeits. Both techniques provide a fast, simple, and accurate way to directly identify among inter-species or intra-species of Penis et testis.

  5. 番木瓜抗病突变抗PRSV基因的RAPD标记分析%RAPD analysis on a gene of resistance to PRSV in Papaya

    Institute of Scientific and Technical Information of China (English)

    黄建昌; 肖艳; 夏有杰

    2009-01-01

    利用随机扩增多态性DNA(Randomly amplified polymorphie DNA,RAPD)分子标记和集团分离分析法(Bulked segregation analysis,BSA)分析技术,对番木瓜(Carica papaya L.)F2代分离群体和抗番木瓜环斑型花叶病毒(Papaya ringspot virus,PRSV)突变体LK-1的抗PRSV基因进行连锁分析,从250个随机引物中筛选出了与抗PPLSV基因相连锁的3个RAPD标记,连锁分析表明,S1366、S7058和S7125与抗PRSV基因(rys)的遗传距离分别为1.9 cM、2.4 cM和12.2 cM,其中S1366和S7058两个标记在抗PRSV基因的-侧,S7125在另-侧.

  6. RAPD-PCR and real-time PCR HRM based genetic variation evaluations of Urtica dioica parts, ecotypes and evaluations of morphotypes in Turkey.

    Science.gov (United States)

    Uzonur, Irem; Akdeniz, Gamze; Katmer, Zeynep; Ersoy, Seyda Karaman

    2013-01-01

    Urtica dioica is an ethnobotanically and medicinally important Complementary and Alternative Medicine (CAM) plant worldwide and in Turkey; 90 % of herbal CAM applications depend on it in Turkey. It has a wide range of habitats in nearly all continents. It is found in all three phytogeographical regions in Turkey (Euro-Siberian, Irano-Turanian, Mediterranean) with high adaptivity to heterogeneous geographies such as climate, soil types and altitudes. This fact in relation to the assessment of chemical constituents of the plant and combining with further genetic and morphological variation data can assist and enhance the works for the utility and reliability of CAM applications in effect and activity of this plant species. In this work we have made some preliminary experiments with novel approaches to reveal the ecotypes and genetic variation of mighty ecotypes of Urtica dioica from different phytogeographical regions of Turkey (Euro-Siberian and Mediterranean). The ecotypes have heterogeneity in both its parts (leaf, stem, root) as revealed by Random Amplified Polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR) using random primers and High-resolution Melt (HRM) analysis using Urtica dioica specific primers and universal chloroplast DNA (cpDNA) primers and morphological traits such as phenolic contents and antioxidant capacities of plants' leaf infusions as used in medicinal applications in Turkey. This work will contribute a lot for the development of molecular markers to detect the genetic variation and heterogeneity of Urtica dioica to further relate with expected phenotypes that are most useful and relevant in CAM applications.

  7. Inbred strains of brine shrimp derived from Artemia franciscana: lineage, RAPD analysis, life span, reproductive traits and mode, adaptation, and tolerance to salinity changes.

    Science.gov (United States)

    Nambu, Fumiko; Tanaka, Shin; Nambu, Ziro

    2007-02-01

    Inbred strains of the brine shrimp were developed from dry dormant cysts of wild-type Artemia franciscana produced in the Great Salt Lake, U.S.A. The established strains were named GSL2, 4, and 7. They were raised in 2% natural sea salt solution at 28 degrees C under a long-day condition, and fed on food sold for Artemia. Ovoviviparous offspring (free-swimming nauplii) in each brood derived from full sib (sister x brother) matings were used for succeeding generations. The ordinal number of the filial generation increased at a rate of ten generations per year. The number was over 60, and the lineage was recorded. Random amplified polymorphic DNA (RAPD) analyses of the inbred strains revealed the uniqueness, homogeneity, and genetic similarity among them. Their life span, the time required to become sexually mature, brood size, mode of reproduction, and adaptation and tolerance to salinity changes were investigated. The inbred strains usually released free-swimming nauplii rather than spawning encysted gastrulae (dormant cysts). On the other hand, the opposite results were obtained from wild-type Artemia under the same conditions. Both adults and nauplii of the inbred strains appeared to be less adaptive and less tolerant to salinity changes compared to those of the wild type. The established inbred strains should provide a wider and deeper scope for Artemia biology in particular, and the life sciences in general.

  8. A RAPD based study revealing a previously unreported wide range of mesophilic and thermophilic spore formers associated with milk powders in China.

    Science.gov (United States)

    Sadiq, Faizan A; Li, Yun; Liu, TongJie; Flint, Steve; Zhang, Guohua; He, GuoQing

    2016-01-18

    Aerobic spore forming bacteria are potential milk powder contaminants and are viewed as indicators of poor quality. A total of 738 bacteria, including both mesophilic and thermophilic, isolated from twenty-five powdered milk samples representative of three types