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
Mario Paredes C
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
Frank Solano Campos
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.
Jaime Eduardo Muñoz Flórez
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.
Muñoz Flórez Jaime Eduardo
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.
Jaime Eduardo Muñoz Flórez; Ana Cruz Morillo Coronado; Yacenia Morillo Coronado
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 difere...
Ana Cruz Morillo Coronado
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
JAVIER RODRÍGUEZ; SIGNED PRIETO; ESPERANZA FAJARDO; CATALINA CORREA; FREDY LÓPEZ; JAIRO CASTRO; YOLANDA SORACIPA
La obesidad y el sobrepeso constituyen problemas de salud pública tanto a nivel mundial como en América Latina. La teoría de la probabilidad y la caminata al azar probabilista han sido base para el desarrollo de predicciones de la dinámica de diferentes epidemias. En este trabajo se estudió el crecimiento de la obesidad en cuatro países con base en la caminata al azar, para confirmar la capacidad de la metodología para describir y predecir esta dinámica específica, y se desarrollaron simulaci...
Julieta Torres Jaramillo; Jaime Eduardo Muñoz; Heiber Cárdenas; Luz ángela álvarez; Juan Diego Palacio
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) p...
Helfgott, Harald Andres
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...
Juan Porfirio Legaria Solano
En este estudio se comparó la eficiencia de dos marcadores, uno bioquímico (proteínas faseolinas) y otro genético (RAPD, polimorfismos en el ADN amplificados al azar) para evaluar la diversidad genética y establecer relaciones filogenéticas entre P. vulgaris, P. lunatus y P. coccineus. El análisis de los patrones de faseolinas permitió detectar polimorfismo entre las especies evaluadas, pero no entre variedades de frijol o individuos de una variedad. Las especies más relacio...
Jaime Eduardo Muñoz Flórez
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
G. CECILIA TORO
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
Viviana Becerra V
Full Text Available Los hongos entomopatógenos son una alternativa atractiva para el control biológico de insectos plagas. En Chile, el Instituto de Investigaciones Agropecuarias (INIA, Centro Regional de Investigación Quilamapu mantiene aproximadamente 400 aislamientos de Beauveria, colectados a través del país. Esta colección ha sido parcialmente clasificada en base a su morfología y a su eficacia como controlador biológico. Sin embargo, es necesario complementar estos estudios con una caracterización a nivel genético-molecular, para determinar el nivel de diversidad genética de Beauveria bassiana e identificar posibles patrones de bandas que permitan dicriminar entre aislamientos. En este estudio, se analizaron 36 aislamientos de B. bassiana provenientes de diversas regiones geográficas del país. El análisis genético se realizó mediante el ADN Polimórfico Amplificado al Azar (RAPD y la Reacción en Cadena de la Polimerasa-Fragmentos de Restricción Polimórficos (PCR-RFLP de los espaciadores internos transcritos (ITS de las secuencias ribosomales del ADN (ITS-rDNA. El análisis de RAPD indicó una alta diversidad genética entre los aislamientos, con un promedio de 43% de similitud. Por otro lado, el análisis de los ITS determinó una menor diversidad, con un 83% de similitud entre aislamientos. La región ITS 1 mostró un mayor número de sitios de restricción que la región ITS 2. Los RAPD fueron más eficientes para discriminar (o identificar entre cepas debido al bajo número de haplotipos detectados con los ITS. Para los dos marcadores utilizados en este estudio, la diversidad genética no estuvo asociada con el origen geográfico de los aislamientos.Entomopathogenic fungi are an attractive alternative for the biological control of insects. In Chile, the Quilamapu Regional Research Center of the National Agricultural Research Institute (INIA has collected about 400 isolates of the genus Beauveria along the country. A partial
Julieta Torres Jaramillo
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
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
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
Michael K. Hole
Full Text Available We report the case of a six-year-old Brazilian girl referred for splenomegaly who first presented with fever, asthenia, and weight loss. Geographical location, clinical exam, and blood laboratories suggested kalaazar. Serology confirmed kala-azar diagnosis, but direct evidence of the parasites was not made. A treatment by meglumine antimoniate is given under hospital surveillance for two weeks. Thereupon, the patient is asymptomatic and all tests are normal.
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
Massive splenomegaly was found in a liver-spleen scan in a young man from Greece. Kala-azar was suggested as a possible cause and was confirmed by culturing Leishmania donovani from the bone marrow aspirate. The differential diagnosis of massive splenomegaly should include kala-azar when a patient has been in an endemic area
Rao, J S; Rahman, S J; Singh, J; Singh, S K
Door to door search during 1991 in 85 villages in Ballia district of Uttar Pradesh revealed 29 sporadic cases of Kala-azar in four villages. Epidemiological investigations indicated indigenous transmission in Phulwaria village (PHC: Dubhar) with 25 cases while the remaining three villages showed four imported cases from the endemic states. No kala-azar cases had been reported in Ballia between 1947 and the present investigation. Phlebotomus arqentipes, the known vector of kala-azar in India, was encountered in 10 PHCs including the four villages having kala-azar cases. Regular vigilance in Ballia and the neighbouring districts in Uttar Pradesh bordering Bihar is suggested in view of indigenous transmission detected in one village with multiple infection in families. The presence of high vector density, ambient environmental factors and absence of regular residual insecticidal spray warrant constant surveillance in Kala-Azar prone areas in Uttar Pradesh. PMID:1344169
Muñoz Flores Jaime Eduardo
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.
Random amplified polymorphic DNA (RAPD) was recently used to assess the variation and phylogenetic relationship of plant material. Some work in this field has been done on sugar beet. The results of a preliminary study to determine whether RAPD can be applied to discriminate between different beetroot (Beta vulgaris L.) breeding material are discussed. 5 refs, 1 fig
Martin Vivodík; Želmíra Balážová; Zdenka Gálová
Castor (Ricinus communis L.) is an important plant for production of industrial oil. The systematic evaluation of the molecular diversity encompassed in castor inbreds or parental lines offers an efficient means of exploiting the heterosis in castor as well as for management of biodiversity. The aim of this work was to detect genetic variability among the set of 30 castor genotypes using 5 RAPD markers. Amplification of genomic DNA of 30 genotypes, using RAPD analysis, yielded 35 fragments, w...
Bittencourt Achiléa; Silva Nancy; Straatmann Andréa; Nunes Victor Luiz Correia; Follador Ivonise; Badaró Roberto
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 leishm...
Deciduous fruit cultivars are traditionally distinguished by means of their phenotypic traits. Since the fruit characteristics are assessed, confirmation of the authenticity of the cultivars can be delayed for as long as 3-4 years. Random amplified polymorphic DNA (RAPD) markers generated by polymerase chain reaction (PCR) have been successfully used to differentiate cultivars in other crops. Therefore, use of RAPD markers was investigated as a means of providing the South African fruit industry with a technique that can rapidly and reliably identify cultivar. 4 refs, 2 figs
Oswaldo P\\u00E1ez; Roberto Valverde; Luis G\\u00F3mez; Arturo Brenes
Para determinar la diversidad genética de Phytophthora infestans en Costa Rica, 62 aislamientos de este patógeno fueron recolectados en plantaciones de papa en las zonas de Cartago, Zarcero, Fraijanes y Heredia durante 1999-2001 y analizados con el uso de RAPDs. Todos los aislamientos fueron previamente evaluados para el tipo de apareamiento y la resistencia al metalaxyl. Se seleccionó 11 imprimadores con los cuales fue posible formar 17 grupos RAPD cuyas frecuen...
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.
After traveling into regions endemic for leishmaniasis all patients presenting with generalized febrile symptoms, pancytopenia, hepatosplenomegaly and symptoms as well as laboratory parameters also seen in autoimmune diseases should be screened for leishmaniasis even after a longer time period. Doctors should bear in mind that especially immunosuppressed patients can present with atypical and abortive symptoms and that in this group of patients immune diagnosis is unreliable. A diagnosis of a lymphoma related only to the spleen should not be made and splenectomy must not be carried out before reliable exclusion of kala azar, using modern diagnostic tools like PCR. Patients should be referred to specialized centers e.g. institutes for tropical medicine and therapy must follow actual guidelines, such as the German guidelines published by the AWMF. PMID:16819655
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.
Yin, Yan-Ni; Chen, Yong-Fang; Li, Shi-Mo; Guo, Jian-Hua
RAPD analysis was used for the taxonomy of plant pathogenic coryneform bacteria, especially for the classification of two new pathogens (Curtobacterium flaccumfaciens pv. basellae pv. nov. and Curtobacterium flaccumfaciens pv. beticola pv. nov.). 20 random primers were screened from 50 ones to detect polymorphism among the total strains used. 80.4% were polymorphic bands among the 225 ones produced. The results of pairwise similarity and UPGMA cluster analysis suggest that the two new pathovars of sugar beet (Beta vulgaris var. saccharifera) and malabar spinach (Basella rubra) are genetically close related with Curtobacterium flacumfaciens, and the minimal similarity coefficient is 0.6511. According to the RAPD analysis and previous research, some newly made taxonomic changes of the plant pathogenic coryneform bacteria are discussed. PMID:16496687
In 1990, a large flood-control project called the Flood Action Plan(FAP) was started in Bangladesh. The FAP built many embankments to divert water during the rainy season. This paper includes two case studies of diseases that are related to flood control in Bangladesh: kala-azar and diarrheal disease. Kala-azar is caused by the protozoan Leishmania donovani and is transmitted to humans by the sandfly Phlebotomus argentipes . It was found that the incidence of kala-azar in embanked areas of no...
Showkat Hussain Ganie
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.
@@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.
Moore, A J; Challen, M P; Warner, P J; Elliott, T J
Cultivars of the white button mushroom Agaricus bisporus are difficult to differentiate, which has made strain protection problematic for this crop species. We have used RAPDs to discriminate between 26 strains of A. bisporus, 24 of which were commercial cultivars, and to characterise the genetic relatedness of these strains. Using 20 primers, 211 RAPD markers were identified and used in hierarchical cluster, patristic distance and parsimony analyses. All strains could be differentiated using the aggregated primer data. Although no one primer could differentiate all 26 strains, several individual primers yielded unique fingerprints for a variety of strains. The greatest differences (up to 28% variation) were observed in comparisons with or between two wild collections of A. bisporus. Quondam cultivars, commercial brown and off-white varieties proved more variable than the widely grown 'hybrid' types. Of the 15 hybrid varieties analysed, only one differed substantially (20% or more variable). The patristic and parsimony analyses both demonstrated the gross similarity of the hybrids, many of which appear to be essentially derived varieties from two original hybrid cultivars. RAPD analyses can assist mushroom strain identification and could play a role in the protection of novel cultivars. PMID:11525623
@@ 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.
A combined methodology of cloned RAPD (random amplification of polymorphic DNA) polymorphic bands and in situ hybridisation to polytene chromosomes is an efficient way to initiate construction of a physical and genetic map of insect disease vectors (Dimopoulos et al. 1996, Mutebi et al. 1997). The studies presented here are the first step in developing this approach in tsetse flies. This technology will be used to support tsetse sterile insect technique (SIT) programmes by providing tools with which population structure and isolation can be assessed and genetic markers that can be used to differentiate released flies from wild flies identified. An added benefit is their possible use in unravelling epidemiological complexity and problems regarding speciation (Besansky et al. 1997). Polytene chromosomes of Diptera have been shown to be excellent material for the study of chromosome structure and function as well as for an understanding of the genetics of natural populations (Lefevre 1976). They provide a means for the accurate mapping of chromosome rearrangements and the precise localisation of genes, using both rearrangement analysis and in situ hybridisation. Previous reports on the cytology of the tsetse flies (Riordan 1968, Maudlin 1970, 1979, Southern et al. 1972, Southern and Pell 1973, Davies and Southern 1976, Southern 1980) have described the basic mitotic karyotype in several Glossina species, and demonstrated the presence of well banded polytene chromosomes in pupal trichogen cells (Southern and Pell 1974, 1981, Pell and Southern 1976). Polytene chromosomes were described for G. austeni Newstead, G. morsitans morsitans Westwood, G. pallidipes Austen and G. fuscipes fuscipes Newstead, but these descriptions are difficult to work with as they are drawings of polytene chromosome elements. In this paper, the photographic chromosome maps of pupal scutellar bristles of G. austeni are presented. They show that these chromosomes can be used with much greater ease
Long-fu ZHU; Xian-long ZHANG; Yi-chun NIE
@@ 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.
de los Ríos Gutiérrez, Iván
Full Text Available This article examines Aristotle’s critical account of the three groups of opinions on chance presented in Physica [Phys.] II 4-6 and considers the question of the identity of the advocates of these ideas. Aristotle confronts three common beliefs according to which: i nothing happens by chance; ii chance is the origin of everything; iii chance is the instrument of a divine dimension inscrutable to the human mind. However, we find no explicit references to any authors. Who are the deniers of chance and who are its proponents? Who identifies chance with divine functions or particular deities? This article aims to outline the debate on the identity of the advocates of these common beliefs and to contribute to its clarification within the frame of the Aristotelian criticism on ancient atomism.En este artículo se examina la exposición crítica de los tres grupos de opiniones sobre el azar que Aristóteles presenta en Physica [Phys.] II 4 y se pregunta por la identidad de sus portavoces. Aristóteles confronta tres creencias establecidas de acuerdo con las cuales: i nada sucede por azar; ii el azar es el origen de todas las cosas; iii el azar es el instrumento un orden divino inescrutable al razonamiento humano. No obstante, en ningún momento encontramos referencias explícitas a uno o varios pensadores. ¿Quiénes son los negadores del azar y quiénes sus defensores? ¿A quién pertenece la identificación del azar con funciones divinas o divinidades singulares? Objetivo del presente artículo es perfilar el debate en torno a identidad de los defensores de las tres posiciones y contribuir, en la medida de lo posible, a su esclarecimiento en el marco de la crítica aristotélica del atomismo antiguo.
Premshanker S. Singh
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
Gilmar E. Martinello
Full Text Available Avaliou-se a utilização de marcadores RAPD para estimar a diversidade em 42 acessos do gênero Abelmoschus e um de Hibiscus. As estimativas das distâncias genéticas foram feitas com base no complemento aritmético do Índice de Jaccard. Foram utilizadas as técnicas de análise multivariada, através de agrupamento hierárquico do vizinho mais próximo e método de Tocher, para estudar os arranjos dos grupos de genótipos, bem como analisar os métodos de agrupamentos empregados. Trinta e um iniciadores foram utilizados para amplificar fragmentos de DNA pela reação de polimerização em cadeia (PCR e foram gerados 103 fragmentos RAPD. O agrupamento hierárquico dos 43 genótipos com base no método do vizinho mais próximo separou os acessos, de modo geral, conforme as espécies botânicas, formando 6 grupos distintos. Isto foi confirmado pela projeção das distâncias genéticas no plano bidimensional, onde o primeiro e maior dos grupos reuniu os acessos de A. esculentus e A. caillei. Por outro lado, o método de Tocher reuniu 90% do germoplasma no grupo I incluindo, neste, os acessos de A. moschatus e A. manihot, além das outras duas espécies anteriores. O método de otimização de Tocher permitiu a formação de apenas 4 grupos de genótipos, mostrando-se coerente apenas em parte à análise de agrupamento hierárquico. Porém, o reagrupamento dos acessos do grupo I de Tocher pelo método hierárquico, revelou a existência de maior heterogeneidade genética no germoplasma estudado.RAPD markers were utilized to estimate the diversity among 42 Abelmoschus and 1 Hibiscus accessions . The genetic distances were based on the arithmetic complement of the Jaccard index. For this purpose we used the multivariate analysis technique by hierarchycal single linkage and the Tocher methods to obtain the genotypes agglomeration as well as to analyze the methods employed. Thirty-one random decamer primers were used do amplify DNA by the polymerase
SAWAZAKI H. E.
Full Text Available Estudou-se, mediante polimorfismo enzimático em gel de poliacrilamida e polimorfismo de DNA com base na amplificação de segmentos de DNA ao acaso, denominado RAPD, a variabilidade genética em algumas espécies e ecótipos de palmeiras dos gêneros Euterpe, Bactris, Elaeis e Syagrus. Os extratos de folhas de mudas dessas palmeiras foram analisados para as isoenzimas de malato desidrogenase (MDH, leucinoaminopeptidase (LAP, glutamato oxaloacetato transaminase (GOT, fosfoglucose isomerase (PGI, fosfoglucose mutase (PGM, fosfatase ácida (ACP, peroxidase (PRX, esterase (EST e para os marcadores RAPD, utilizando os "primers" dos kits A e B da Operon Technologies. Verificou-se grande variabilidade genética interespecífica, comprovada pelos dendrogramas UPGMA, com reconhecimento de híbridos. Foram observadas várias bandas além das referidas pela literatura em gel de amido. Os resultados dos marcadores RAPD comprovaram os das isoenzimas com maior eficácia, pois possibilitaram facilmente a análise de grande número de marcadores genéticos.
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.
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.
Jorge Iván González
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.
ROMÁN, B.; Alfaro, C; Torres, A. M.; MORENO, M. T.; SATOVIC, Z.; PUJADAS, A.; Rubiales, D.
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...
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)
Sankha Koley; Rajesh Kumar Mandal; Sanjiv Choudhary; Arghya Bandyopadhyay
Post-kala-azar dermal leishmaniasis (PKDL) is an unusual dermatosis occurring following an attack of visceral leishmaniasis (VL). There are only few reports of PKDL after successful treatment with miltefosine. We report two cases of PKDL that developed after successful treatment of VL with miltefosine.
Ismail, A; El Hassan, A M; Kemp, K;
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 the immunol......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...... of macrophages, lymphocytes, and plasma cells. In patients who had high interferon-gamma (IFNgamma) responses to Leishmania antigen in vitro, compact epithelioid granulomas were formed. The inflammatory cells were mainly CD3(+) and interleukin-10 (IL10) was the most prominent cytokine in the lesions. However......, IFNgamma was found in all and IL4 in most lesions, in varying amounts. PBMCs from all patients responded to Leishmania antigen by IFNgamma production or proliferation. The results indicate that PKDL develops as a result of an influx of immunocompetent cells into skin, which harbours parasites...
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
MARIA VITÓRIA CECCHETTI GOTTARDI
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
Nielsen, Karen L; Godfrey, Paul A; Stegger, Marc;
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...
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
Ana Cruz Morillo Coronado
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”.
Somaclonal variation in 13 R2 families and the FR-13A parent was investigated at the DNA level using the random, amplified polymorphic DNA (RAPD) method via polymerase chain reaction. Thirty-four random primers were employed to amplify the DNA segments. Six 10-mer primers revealed a total of 67 scorable bands, all of which were polymorphic across the 70 individual plants (five plants and 65 R2 lines). On the basis of the simple matching coefficient of the RAPD banding patterns, all the somaclonal R2 families showed significant dissimilarity with the FR-13A parent. This study demonstrated that molecular makers such as RAPD are useful in revealing changes in DNA sequence not readily seen in morphological or physiological screening. 1 fig
RAPDs were quite efficient in bringing out the diversity at DNA level among non-edible legumes viz., Acacia nilotica, Adenanthera pavonina, Prosopis juliflora, Pithecolobium dulce, Clitoria ternatea and Pongamia pinnata. The RAPD primer index reveals the information content of the RAPD primer per se. Of the 82 primers tested, OPE 8, OPI 6, OPL 2, OPL 16, OPI 18, OPI 13, OPI 14, OPP 1, OPE 20 and OPI 4 with comparatively higher primer index were more informative and can be used for further DNA finger printing and population studies in tree legumes. CTAB protocol was found to be superior in isolating genomic DNA of good quality. The 260/280 ratios varied between 1.70 and 2.09. Though the genomic DNA isolated by potassium acetate method was found to be intact in 0.8% agarose gel, the yield was significantly lower than the modified CTAB method. (author)
V. M. Shinde
Full Text Available In this study, the RAPD (Random Amplified Polymorphic DNA technique was employed for determination of the components in an Ayurvedic herbal prescription, Rasayana Churna. One-hundred-and-twenty decamer oligonucleotide primers were screened in the RAPD analysis to identify three Ayurvedic medicines, dried stem of Tinospora cordifolia, dried fruit of Emblica officinalis and dried fruit of Tribulus terestris, the Ayurvedic prescription. Primer OPC-6 simultaneously generated three distinct amplicons, each specific to one component. The marker with 600 bp is specific to Tinospora cordifolia; the marker 500 bp is specific to Emblica officinalis and the remaining marker >1000 bp was present in Tribulus terestris. Presence of three herbal medicines was determined when RAPD reaction with OPC-6 was performed. The technique was proved to contribute to the identification of components in Ayurvedic herbal preparation and thus helping to serve as a complementary tool for quality control.
WU Xiaoqin; XIONG Dabin; WANG Yu
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.
focus on the conserved ribosomal regions for phylogenetic diversity studies are those that amplify up random genomic sequences. The two most common methods are random amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP)
Xue Xian; Lin Shanzhi; Zhang Zhixiang
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.
LI Yu-Feng; HUANG Qun-Ce; LIANG Yun-Zhang; YU Zeng-Liang
Germination capacity of alfalfa seeds under low energy N+ implantation manifests oscillations goingdown with dose strength. From analyzing alfalfa genome DNA under low energy N+ implantation by RAPD (RandomAmplified Polymorphous DNA), it is recommended that 30 polymorphic DNA fragments be amplified with 8 primersin total 100 primers, and fluorescence intensity of the identical DNA fragments amplified by RAPD is different be-tween CK and treatments. Number of different polymorphic DNA fragments between treatment and CK via N+ im-plantation manifests going up with dose strength.
WANG Jingang; ZHAO Hongkun; GONG Shufang; CHE Daidi
RAPD analysis was applied in studying the classification and genetic relationship of 12 cultivars of Gladiolus hybridus Hort. Twenty-six arbitrary primers screened from 80 primers were used for further PCR and diversity analysis. A total of 164 RAPD sites were detected with a mean of 6.31 fragments amplified for each primer. A total of 151 polymorphic DNA fragments were detected among all the 164 amplified fragments, which accounted for a high level of 92.8% of all and could be used for identification of different cultivars. The result revealed that the germplasm resource of Gladiolus hybridus Hort. cultivar had a narrow genetic base on molecular level.
BAI Heine; NIU De; WANG Lijuan
Random amplified polymorphic DNA (RAPD) was used to study the diversity of seven E. pulcherrima with different color leaves. Aight 10 bp primers selected from 30 arbitrary primers were applied for amplifying the Euphorbia pulcherrima DNA. Total 62 bands were produced, among which 22 bands (35.48%) were polymorphic. The average numbers of polymorphic DNA bands amplified by each primer were 7.8. Cluster analysis results showed that the genetic background of seven E. pulcherrima was relatively narrow and they had the higher similarity coefficient, which indicated that these seven strains had a near relationship, and that RAPD marking technology may be the effective means for E. pulcherrima classification.
Germination capacity of alfalfa seeds under low energy N+ implantation manifests oscillations going down with dose strength. From analyzing alfalfa genome DNA under low energy N+ implantation by RAPD (Random Amplified Polymorphous DNA), it is recommended that 30 polymorphic DNA fragments be amplified with 8 primers in total 100 primers, and fluorescence intensity of the identical DNA fragment amplified by RAPD is different between CK and treatments. Number of different polymorphic DNA fragments between treatment and CK via N+ implantation manifests going up with dose strength
Adams, Emily R.; Inge Versteeg; Leeflang, Mariska M. G.
Identification of post-kala-azar dermal leishmaniasis (PKDL) is important due to the long and toxic treatment and the fact that PKDL patients may serve as a reservoir for visceral leishmaniasis (VL). We summarized the published literature about the accuracy of diagnostic tests for PKDL. We searched Medline for eligible studies investigating the diagnostic accuracy of any test for PKDL. Study quality was assessed using QUADAS-2. Data were extracted from 21 articles including 43 separate studie...
Ghalib, Hashim; Modabber, Farrokh
Background Post kala azar dermal leishmaniasis (PKDL) is a disease that appears after treatment of visceral leishmaniasis (VL). The highest incidence of PKDL in the world is in Sudan. Many patients heal spontaneously within 6 months but those who don't are difficult to treat, often requiring months of daily injections. These patients harbour parasite in their skin and are believed to be a source of infection and possibly epidemics. Present treatment modalities of PKDL are inadequate and impra...
Belay, A Dejenie; Asafa, Y; Mesure, J; Davidson, R. N.
The first two patients to be treated with miltefosine for post-kala-azar dermal leishmaniasis (PKDL) are reported. One was a 26-year-old Ethiopian man who had been treated with sodium stibogluconate, for relapsing visceral leishmaniasis (VL), four times between August 2002 and March 2004. In January 2004 this patient was found to be seropositive for HIV and began antiretroviral treatment with stavudine, lamivudine and nevirapine. Five months later he developed clinical PKDL, with extensive cu...
Full Text Available Introduction & Objective: Visceral Leishmaniasis (kala-azar is the most important endemic disease in Northwestern Iran, particularly in Ardabil province. This study aimed to review the seroepidemiological studies which have been performed in Ardabil province during 1986-2009. Materials & Methods: In this descriptive analytical study, studies which have been carried out from 1986 through 2009 in Northwestern Iran about clinical, diagnostic and epidemiological features of Kala azar, using DAT, were reviewed. Collected data were analyzed using the SPSS software. Results: in total, 2703 of human visceral leishmaniasis were detected by direct agglutination test (DAT in Ardabil province, 1787 (66.1% of them were from Meshkin-shahr district, 837 (31% cases were from Moghan district, and 79 (2.9% cases were from Ardabil district. Ninety eight percent of the cases were under 10 years old while only 0.5% of the VL cases were ≥20 years old and 17% of them were under 1 year of age. Conclusion: Currently Kala-Azar is the most important endemic disease in Northwestern Iran, particularly in Ardabil province. Anti-Leishmania antibodies at the titers of ≥1:3200 using DAT along with clinical signs including fever, anemia and hepatosplenomegaly are considered as active visceral leishmaniasis. DAT antibody titer of 1/800 and lower and absent of clinical signs is considered as negative VL.
Lessells, C.M.; Mateman, A.C.
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
Full Text Available Genetic polymorphism and relationships among 30 commercial varieties of Bangladeshi aromatic rice (Oryza sativa L. were established using random amplified polymorphic DNA (RAPD primers. Out of fifty 10-mer RAPD primers screened initially, four were chosen and used in a comparative analysis of different varieties of indigenous Bangladeshi aromatic rice. Of the 33 total RAPD fragments amplified, 7 (21.21% were found to be shared by individuals of all eight varieties. The remaining 26 fragments were found to be polymorphic (78.79%. Pair-wise estimates of similarity ranged from 0.101 to 0.911. Highest genetic diversity was determined between Radhunipagol and Dubsail varieties (0.911. The amount of genetic diversity within aromatic rice germplasm was quite high as determined by the genetic similarity coefficients between varieties. Genetic similarities obtained from RAPD data were also used to create a cluster diagram. Cluster analysis using an un-weighted pair-group method with arithmetic averages (UPGMA was used to group the varieties and the 30 aromatic rice varieties were grouped into 6 clusters where cluster I includes the maximum number of varieties (9. Cluster VI includes minimum number of varieties (2. This Study offered a rapid and reliable method for the estimation of variability between different varieties which could be utilized by the breeders for further improvement of the local aromatic rice varieties.
Bhunia Gouri Sankar
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
ZHENG Dao-jun; LIANG Yuan-fa; LIU Gou-min; YAN Dong-hai; LINGHU Chang-di; TIAN Yong-hui
The random amplified polymorphic DNA marker (RAPD) was applied to detect the genetic relationships and diversity among 21 germplasm materials of Kudingcha species in Oleaceae, which involved 8 species, i.e., Ligustrum robustum (Roxb.) Blume, L. henryi Hemsl., L. japonicum Thunb, L. japonicum Thunb. vat. pubscens Koidz, L. luciduM Ait., L. pedunculare Rehd, Osmanthus masumuranus Hayata, and L. delavayanm Hariot. 20 RAPD primers selected were applied for the amplification on the 21 germplasm materials mentioned above. 427 bands were obtained, and the percentage of polymorphic bands (PPB) was 97.7%. The genetic similarity coefficients (GS) ranged from 0.1522 to 0.8322 with an average of 0.5466. There was a significant genetic difference among germplasm materials of Kudingcha species in Oleaceae, and UPGMA cluster based on the GS of RAPD could distinguish all test germplasm materials clearly and indicated the relationship of the 8 species mentioned above, all of which indicated that RAPD markers could be used for the studies of genetic diversity and relationship and classification of germplasm resources of Kudingcha species in Oleaceae. Analysis results of RAPD showed that L. japonicum Thunb. var. pubscens Koidz has closer genetic relationship with L. pedunculare Rehd and further genetic relationship with L. japonicum Thunb. among all tested species. The authors suggest that further research is needed to study whether L. japonicum Thunb. var. pubscens Koidz should be classified into a variata of L. japonicum Thunb, or should be considered as an independent species. The analysis results supported that L. pururascens Y. C. Yang should be combined into L. robustum (Roxb.) Biume.
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.
Gasim, S; Theander, T G; ElHassan, A M
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 the...... 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....
Uranw, Surendra Kumar
Visceral leishmaniasis (VL) or kala-azar is a parasitic infectious disease that is fatal if left untreated. Two types of Leishmania species are causal agents of VL: Leishmania infantum and Leishmania donovani. VL caused by L.infantum is a zoonosis and is endemic in countries around the Mediterranean basin and in Latin-America. VL caused by L. donovani is assumed to be an anthroponosis and is endemic in East-Africa and the Indian subcontinent.VL is considered as a major public health problem i...
Daniela L Leite
Full Text Available Em plantas alógamas, as cultivares constituem uma fonte de variação genética planta-a-planta no qual o melhorista pode obter novos materiais. A variabilidade genética entre indivíduos ou populações tem sido medida através de marcadores moleculares. O objetivo deste trabalho foi avaliar molecularmente, através da análise de RAPD (Random Amplified Polimorphic DNA, a diversidade genética existente em um grupo de cultivares de cebola recomendadas para o cultivo na Região Sul do Brasil. Foram analisados 11 iniciadores em DNAs provenientes de um total de 90 plantas individuais oriundas das cultivares Baia Periforme, Bola Precoce, BRS Cascata, Crioula, Primavera e Roxa. As plantas foram genotipadas através dos produtos visualizados em gel de agarose e uma matriz de presença/ausência de bandas foi utilizada nas análises de similaridade genética (coeficiente de Jaccard e de agrupamento pelo método UPGMA. Os 11 iniciadores geraram 140 (86,4% bandas polimórficas. O número de fragmentos por iniciador variou de 10 (UBC105 até 18 (OPA10; OPC11; OPI1 com uma média de 15 bandas por iniciador. A técnica de RAPD mostrou-se eficiente na caracterização molecular, permitindo a formação de grupos de cultivares de acordo com a sua população de origem, com uma similaridade média de 63%. Conclui-se que as cultivares de cebola recomendadas para a Região Sul do Brasil, apresentam divergência genética, com potencial de serem exploradas por melhoristas, no desenvolvimento de genótipos superiores.In alogam plants, the cultivars constitute a source of plant-to-plant genetic variation from which the breeder can obtain new materials. Genetic variability among individuals or populations has been measured through molecular markers. The objective of this work was to evaluate the genetic diversity in a group of onion cultivars recommended for cultivation in the Southern Region of Brazil by RAPD (Random Amplified Polimorphic DNA analysis. Eleven
Ashish G Vala1*, R.S.Fougat1 and G.C.Jadeja
Full Text Available Genetic variability of 15 sets of Plantago ovata Forsk. studied using 11 arbitrary oligonucleotide primers. Among the 90 DNAfragments produced 71 fragments were found to be polymorphic. The mean number of polymorphic bands per primer among 15Plantago ovata genotypes was 6.45 . The higher polymorphism (90.00 % was exhibited by primer OPF-17, while the lowerpolymorphism (60.00 % was detected by OPF-2. The genetic similarity matrix from RAPD data for 15 genotypes was calculatedbased on Jaccard’s coefficients of similarity ranged from 0.45 to 0.80. UPGMA cluster analysis reveals that the 15 genotypeswere clustered in to three clusters. Genetically distinct genotypes identified using RAPD markers could be potential sources ofgermplasm for Isabgol improvement.
Full Text Available Anatomical and Random Amplified Polymorphic DNA (RAPD analysis of two typical populations of Salicornia europaea from Montenegro and Greece (Lesvos, one typical population of S. ramosissima from Spain and one population that belongs to the Salicornia genus from Serbia, was undertaken to develop a new strategy for identifying Salicornia plants. Anatomical variability and differentiation were examined using Principal Component Analysis (PCA and Multivariate Discriminant Function Analysis (MDA. On the basis of the anatomical measurements, the four populations were classified into three groups: one joining the plants from Serbia and Spain, one comprising the Montenegrin group and one comprising the Lesvos group. RAPD analysis indicated that populations from Spain and Serbia were closely related to each other and the Lesvos group was quite different from all the other investigated populations. These results opened up the possibility that the specimens from Serbia belonged to S. ramosissima and not to S. europaea, as reported previously.
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.
Stegniĭ, V N; Chudinova, Iu V; Salina, E A
Genetic polymorphism in varieties and hybrids of cultivated flax (Linum usitatissimum L.) has been investigated by RAPD-PCR. Analysis with 15 primers has revealed varietal specificity and hybrid inheritance of RAPD alleles. This allows genetic certification of the original varieties and their hybrids for breeding purposes. Polymorphic amplification products were obtained in RAPD analysis of DNA from two cultivated flax varieties with the use of 10-11 nucleotide primers. PMID:11094749
Salinity, which is critical in determining the growth and development of plants, is a major problem affecting ever-increasing areas throughout the world. A salt tolerant rice mutant (M-20) was obtained from accession 77-170 (Oryza sativa) through EMS mutagenesis and selection in vitro. The use of 220 10-mer RAPD primers allowed the identification of a new molecular marker, whose genetic distance from a salt tolerance gene is about 16.4 cM. (author)
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
Mkada-Driss, Imen; Lahmadi, Ramzi; Chakroun, Ahmed S; Talbi, Chiraz; Guerbouj, Souheila; Driss, Mehdi; Elamine, Elwaleed M; Cupolillo, Elisa; Mukhtar, Moawia M; Guizani, Ikram
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, to develop
Mkada-Driss, Imen; Lahmadi, Ramzi; Chakroun, Ahmed S; Talbi, Chiraz; Guerbouj, Souheila; Driss, Mehdi; Elamine, Elwaleed M; Cupolillo, Elisa; Mukhtar, Moawia M; Guizani, Ikram
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 characte...
Tímea Kuťka Hlozáková; Zdenka Gálová; Edita Gregová; Martin Vivodík; Želmíra Balážová; Dana Miháliková
Wheat (Triticum aestivum L.) is one of the main crops for human nutrition. The genetic variability of grown wheat has been reduced by modern agronomic practices, which inturn prompted the importance of search for species that could be useful as a genepool for the improving of flour quality for human consumption or for other industrial uses. Therefore, the aim of this study was to analyze the genetic diversity among 24 European wheat genotypes based on Random Amplified Polymorphism (RAPD) mar...
Milić Dubravka; Luković Jadranka; Đan Mihajla; Zorić Lana; Obreht Dragana; Veselić Sanja; Anačkov G.; Petanidou Theodora
Anatomical and Random Amplified Polymorphic DNA (RAPD) analysis of two typical populations of Salicornia europaea from Montenegro and Greece (Lesvos), one typical population of S. ramosissima from Spain and one population that belongs to the Salicornia genus from Serbia, was undertaken to develop a new strategy for identifying Salicornia plants. Anatomical variability and differentiation were examined using Principal Component Analysis (PCA) and Multivariate Discriminant Function Analys...
Welington Luiz Araújo; Derlene Attili de Angellis; João Lúcio de Azevedo
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 d...
YANHUAJUN; SILANDAI; 等
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.
Marouelli, L P; Inglis, P W; Ferreira, M A; Buso, G S C
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. PMID:20645261
Guo, H R; Zhang, S C; Tong, S L; Xiang, J H
We tested the applicability of the random amplified polymorphic deoxyribonucleic acid (RAPD) analysis for identification of three marine fish cell lines FG, SPH, and RSBF, and as a possible tool to detect cross-contamination. Sixty commercial 10-mer RAPD primers were tested on the cell lines and on samples collected from individual fish. The results obtained showed that the cell lines could be identified to the correspondent species on the basis of identical patterns produced by 35-48% of the primers tested; the total mean similarity indices for cell lines versus correspondent species of individual fish ranged from 0.825 to 0.851, indicating the existence of genetic variation in these cell lines in relation to the species of their origin. Also, four primers, which gave a monomorphic band pattern within species/line, but different among the species/line, were obtained. These primers can be useful for identification of these cell lines and for characterization of the genetic variation of these cell lines in relation to the species of their origin. This supported the use of RAPD analysis as an effective tool in species identification and cross-contamination test among different cell lines. PMID:11573817
Thakur, Chandeshwar P.; Kumar, A.(State University of New York at Buffalo, Buffalo, USA); Mitra, Dipendra K.; Roy, Ambak; Sinha, Arun Kumar; Ranjan, Alok
Complications of amphotericin B limit its wide application in the treatment of patients with kala-azar. This study was undertaken with an aim to minimize anti-renal complications and severe rigor in course of treatment with this drug. Parasitologically confirmed kala-azar cases (n = 230) were randomized equally into two groups: a control group received amphotericin B only at a dose of 1 mg/kg of body weight/day for 20 days and a patient (test) group received 500 mL of physiologic saline and 3...
MUKHERJEE, Ashutosh; SIKDAR, Biswanath; Ghosh, Biplab; Banerjee, Anuradha; GHOSH, Enakshi; BHATTACHARYA, Moimee; ROY, Satyesh Chandra
RAPD and ISSR markers have been used to reveal the genetic diversity and phylogenetic analysis of some economically important species of Allium. Two plants related to Allium were also included for better understanding of the phylogeny. ISSR showed more polymorphism within A. cepa and A. sativum, while RAPD showed more polymorphism within A. porrum. Overall, RAPD revealed more intraspecific diversity than ISSR, while ISSR showed more interspecific diversity than RAPD. This showed the usefulnes...
Colauto Nelson Barros
Full Text Available The genetic divergence of five isolates of Agaricus blazei was determined based on RAPD data. Results indicate that there is little genetic variability among the commercialized strains and that RAPD is a feasible and low cost technique that can be used to characterize this fungus.
Colauto Nelson Barros; Dias Eustáquio Souza; Gimenes Marcos Aparecido; Eira Augusto Ferreira da
The genetic divergence of five isolates of Agaricus blazei was determined based on RAPD data. Results indicate that there is little genetic variability among the commercialized strains and that RAPD is a feasible and low cost technique that can be used to characterize this fungus.
Kappe, A.L.; van de Zande, L.; Vedder, E.J.; Bijlsma, R.; van Delden, Wilke
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.
Pinto-Carnide, Olinda; MARTÍN, JUAN PEDRO; Leal, Fernanda; Castro, Isaura; Guedes-Pinto, Henrique; Oriz, Jesús María
Twelve grapevine cultivars from northern Portugal were characterized with RAPD and microsatellites. Nine primers were used in the RAPD analysis; 8 of the varieties showed monotypic patterns. With the 6 microsatellite loci a total of 38 alleles was obtained. Relationships among the studied varieties were observed. Both types of molecular markers have proved useful for identification. Existence of synonymies is discussed.
Genetic relatedness of gray-tailed voles (Microtus canicaudus) was determined by random amplified polymorphic DNA (RAPD). This work is the first reported use of the RAPD method for pedigree analysis of M. canicaudus and demonstrates the feasibility of RAPD for assessing paternity...
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
Rouf, M A; Rahman, M E; Islam, M N; Islam, M N; Ferdous, N N; Hossain, M A
In Bangladesh, the total population at risk for kala-azar exceeds 20 million (18%) living in 88 Thana (19%) of 27 districts (42%). A confirmatory diagnosis of visceral leishmaniasis (kala-azar) is done by demonstration of the parasite (LD body) in organ aspirates or tissue biopsy sample, an invasive procedure with relatively low sensitivity. To assess the diagnostic usefulness of ICT for antibody against the leishmanial antigen rK39 & its feasibility for use under field conditions (rural areas). An experimental study conducted during January, 2003 to July, 2003 in pediatrics department of MMCH including 60 confirmedly diagnosed KA cases & 60 controls having diseases other than KA. One drop of peripheral blood is applied to the nitrocellulose strip & 3 drops of test buffer is added to the dried blood. Observing 2 visible bands indicates presence of IgG anti-K39. The rK39 strip test was positive in 57 out of 60 confirmed KA case diagnosed by LD body demonstration in splenic or bone marrow aspirate. The estimated sensitivity was 95%. One control diagnosed as other than KA had positive strip test but negative aspirate smear. The estimated specificity of the strip test was 98.3%. The predictive value of a positive result is 98.3% & that of a negative result is 93.5%. rK39 strip test is highly sensitive & specific in our situation & it can be used as a simple & the best method for diagnosis of KA in rural areas. PMID:19377416
Welington Luiz Araújo
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.
SHAO Wan-fang; PANG Rui-hua; DUAN Hong-xing; WANG Ping-sheng; XU Mei; ZHANG Ya-ping; LI Jia-hua
RAPD assessment on genetic variations of 45 tea trees in Yunnan was carried out. Eight primers selected from 40 random primers were used to amplify 45 tea samples, and a total of 95 DNA bands were amplified, of which 90 (94.7 %) were polymorphism. The average number of DNA bands amplified by each primer was 11.5. Based on the results of UPGMA cluster analysis of 95 DNA bands amplified by 8 primers,all the tested materials could be classified into 7 groups including 5 complex groups and 2 simple groups, which was basically identical with morphological classification. In addition, there were some speciations in 2 simple groups.
Adrise Medeiros Nunes
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
Patrícia Pereira Pinto
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.
Full Text Available Grevillea (Proteaceae is native to Australia and is known as garden plant. The flowers have high diversity colors, which range over orange, yellow, pink, red and green. It is a large genus, consisted of 357 species. This genus is economically important and there are hundreds of Grevillea hybrids with beautiful color and shape of flower. Information of the genetic relationships of Grevillea hybrids is not available. Ten Grevillea hybrids were used in this study to evaluate the genetic relationships between hybrids and with their parents. PCR-RAPD technique was employed in this study with seven RAPD primers. The dendrogram of Grevillea hybrids was performed using Neighbor-Joining analysis based on genetic distance. The analysis revealed that ‘Moonlight’, ‘Caloundra Gem’, and Little Pink Willie’ were clustered with Grevillea banksii. The three hybrids were hybrid of G. banksii and other Grevillea species. ‘Superb’ was closely related to ‘Robyn Gordon’ as they are hybrid of G. banksii and G. bipinnatifida.
Identification of aluminum (Al) responsive genes is of great importance in illuminating the molecular mechanism of plant Al response. In this present study, we preliminarily identified several genes that possibly involved in Al-response by cDNA based random amplified polymorphic DNA (cDNA-RAPD) method from Al tolerant/sensitive soybean cultivars exposed to 0 or 50 microM Al3+ solutions for two days. Totally one hundred random primers were used to identify the differentially expressed genes; however, only two primers generated eight stable PCR products. The eight gene fragments were cloned and sequenced, then compared with NCBI gene bank. We subsequently verified the expression profiles of these eight genes by real time quantitative PCR (RT-qPCR) and found that two genes were significantly up-regulated after Al treatment for 24, 48 and 72h. One gene, encoding nucleotide-diphospho-sugar transferase which is essential for polysaccharide synthesis and another gene, encoding polygalacturonase inhibiting protein which exerts its role in terms of inhibiting polysaccharide hydrolysis, suggesting the possibility that they might cooperate in response to Al stress through the modification of cell wall components. These findings provided valuable candidate genes for further study on the molecular mechanisms in plant Al tolerance. (author)
RAPD and ISSR markers were used to analyse intra and inter-specific variability of 16 A. caudatus, 18 A. cruentus and 21 A. hypochondriacus accessions. The potential of both approaches was evaluated using three random and three microsatellite primers amplifying in total of 1126 (RAPD), respectively 1013 (ISSR) scorable fragments. Similarity values among accessions of three Amaranthus species ranged from 0.00 to 1.00 in both types of markers. Based on the intra-specific variability the polymorphism percentage varied from 70 to 100% (RAPD) or from 90 to 100% (ISSR) respectively. Resolving power (Rp) of ISSR primers have been higher (5.28 in average) in comparison to RAPD primers (4.84 in average). Percentage of distinguished accessions by ISSR and RAPD primers ranged from 29 to 89% (based on primer's type) and from 19 to 72%, respectively. Cluster analysis based on RAPD and ISSR data has shown the individual species separation except of three accessions. Two of A. caudatus genotypes originated from India clustered with A. hypochondriacus accessions and one of A. hypochondriacus genotype originated from Nepal clustered with A. cruentus accessions. This study has demonstrated, that a single primer marker systems as RAPD and ISSR are able to generate a sufficient level of informative characters for intra and inter-specific analysis of Amaranthus genus. (author)
Mohamed A. Nagaty
Full Text Available Problem statement: Peach genotypes from the Kingdom of Saudi Arabia were not adequately characterized at the biological and molecular levels. This study was carried out to characterize nine peach cultivars growing in Taif (KSA at the biological and molecular levels. Approach: For the nine peach cultivars studied, flowering and fruiting indices were determined and eight, 10-base primers were used to amplify DNA from each cultivar by using Randomly Amplified Polymorphic DNA (RAPD technology. RAPD profiles (i.e., cultivars and RAPD amplicons (i.e., markers were biclustered by using appropriate software and the RAPD biclusters were viewed as a heatmap in order to improve the display of RAPD markers. Results: The different RAPD primers produced a total of 114 fragments (amplicons, of which 63 were polymorphic among the studied cultivars. The RAPD fingerprinting results confirmed the data obtained from the morphological analyses and allowed for estimating the genetic relatedness among the studied peach cultivars. Moreover, the data indicated that certain molecular markers might be associated with certain commercial characteristics. Conclusion/Recommendations: Future studies on the association(s of the selected molecular markers with fruiting characteristics should allow for new gene discovery, molecular breeding, proper biodiversity assessment and better conservation of germplasm resources.
Tigano Myrian Silvana
Full Text Available The genetic variability of 22 isolates of the fungi Alternaria alternata and A. cassiae, obtained from Senna obtusifolia, was studied by RAPD analysis. A total of 491 scorable bands were produced with the use of 28 primers. Cluster analysis based on similarities computed from RAPD markers showed two distinct genetic groups of isolates related to both species. RAPD analysis proved to be an efficient method for detecting genetic variability of A. cassiae and A. alternata isolates occurring in S. obtusifolia, and also for distinguishing Alternaria species.
Tímea Kuťka Hlozáková
Full Text Available Wheat (Triticum aestivum L. is one of the main crops for human nutrition. The genetic variability of grown wheat has been reduced by modern agronomic practices, which inturn prompted the importance of search for species that could be useful as a genepool for the improving of flour quality for human consumption or for other industrial uses. Therefore, the aim of this study was to analyze the genetic diversity among 24 European wheat genotypes based on Random Amplified Polymorphism (RAPD markers. A total of 29 DNA fragments were amplified with an average 4.83 polymorphic fragments per primer. The primer producing the most polymorphic fragments was SIGMA-D-P, where 7 polymorphic amplification products were detected. The lowest number of amplified fragments (3 was detected by using the primer OPB-08. The size of amplified products varied between 300 bp (OPE-07 to 3000 bp (SIGMA-D-P. The diversity index (DI of the applied RAPD markers ranged from 0.528 (OPB-07 to 0.809 (SIGMA-D-P with an average of 0.721. The polymorphism information content (PIC of the markers varied from 0.469 (OPB-07 to 0.798 (SIGMA-D-P with an average 0.692. Probability of identity (PI was low ranged from 0.009 (SIGMA-D-P to 0.165 (OPB-07 with an avarage 0.043. The dendrogram based on hierarchical cluster analysis using UPGMA algorithm was prepared. Within the dendrogram was separated the unique genotype Insegrain (FRA from the rest of 23 genotypes which were further subdivided into two subclusters. In the first subclaster were grouped 13 genotypes and the second subcluster involved 10 genotypes. The first subcluster also included the genotype Bagou from France, in which were detected novel high - molecular - weight glutenin subunits using SDS-PAGE. Using 6 RAPD markers only two wheat genotypes have not been distinguished. Through that the information about genetic similarity and differences will be helpful to avoid any possibility of elite germplasm becoming genetically uniform
RAPD markers were utilized to identify the genetic differences and the genetic relationship between 8 safflower genotypes i.e. seven induced mutants namely Mut 1 H, Mut 2 H2 , Mut3, Mut4, Mut 5 , Mut6, Mut 7 and the parental variety Giza 1. Ten arbitrary primers were used; different primers generated polymorphic RAPD profiles. The number of amplified DNA amplicons across the ten primers ranged from seven amplicons for the primer OBC-18 to 17 amplicons for the primersOPA-03 and OPA-04. However the number of polymorphic amplicons ranged from 1 for the primer OPB-3 to 14 amplicons for the primers OPA-03 and OPA-17. The percentage of polymorphism ranged from 9.09 % for the primer OPB- 03 to 100% for the primer OPC-17.The highest genetic similarity (94%) was found between Mut 4 and Mut 7 and the lowest (79.0%) was found between Mut 1 and Giza 1. Seventeen positive and four negative unique RAPD markers were identified across the 8 safflower genotypes. The parent Giza 1 was characterized by one positive unique marker amplified by OPA-03 primer at the molecular weight of 2000 bp as well as, two negative unique markers generated by the OPB-6 and OPB-5 primers at the molecular weights of 1150 and 800 bp., respectively. The mutant 1 showed highest number of positive unique markers (8) generated by OPA-3 primer at the molecular weights of 1400, 800 ,700 and 600 bp, OPB-04 at the molecular weight 2000 bp., OPB-06 primers at the molecular weight of 900 bp., OPB-05 primer at the molecular weight of 500 bp., and OPA-04 primer at the molecular weight of 600 bp. Mut 2 was identified by two positive unique markers generated by the OPB-05 and OPA-03 primers at the molecular weights of 1500 and 500 bp respectively, However the Mut 3 was characterized by one positive unique marker amplified by OPC-17 primer at the molecular weight 550 bp., there is no unique number was found to characterize the mutant 4. The Mut 5 identified by one positive uniquemarker generated by OPA-04 Primer at the
Sreenivas, Gannavaram; N. A. Ansari; Kataria, Joginder; Salotra, Poonam
A nested PCR assay to detect parasite DNA in slit aspirates from skin lesions of patients with post-kala-azar dermal lesihmaniasis (PKDL) is described. PCR results were positive in 27 of 29 (93%) samples by nested PCR assay, while only 20 of 29 (69%) were positive in a primary PCR assay. The nested PCR assay allowed reliable diagnosis of PKDL in a noninvasive manner.
Emily R. Adams
Full Text Available Identification of post-kala-azar dermal leishmaniasis (PKDL is important due to the long and toxic treatment and the fact that PKDL patients may serve as a reservoir for visceral leishmaniasis (VL. We summarized the published literature about the accuracy of diagnostic tests for PKDL. We searched Medline for eligible studies investigating the diagnostic accuracy of any test for PKDL. Study quality was assessed using QUADAS-2. Data were extracted from 21 articles including 43 separate studies. Twenty-seven studies evaluated serological tests (rK39 dipstick, ELISA, DAT, and leishmanin tests, six studies molecular tests, eight microscopy, and two cultures. Only a few of these studies reported a valid estimate of diagnostic accuracy, as most were case-control designs or used a reference standard with low sensitivity. The included studies were very heterogeneous, for example, due to a large variety of reference standards used. Hence, no summary estimates of sensitivity or specificity could be made. We recommend well-designed diagnostic accuracy trials that evaluate, side-by-side, all currently available diagnostics, including clinical symptoms, serological, antigen, molecular, and parasitological tests and possible use of statistical modelling to evaluate diagnostics when there is no suitable gold standard.
Adams, Emily R; Versteeg, Inge; Leeflang, Mariska M G
Identification of post-kala-azar dermal leishmaniasis (PKDL) is important due to the long and toxic treatment and the fact that PKDL patients may serve as a reservoir for visceral leishmaniasis (VL). We summarized the published literature about the accuracy of diagnostic tests for PKDL. We searched Medline for eligible studies investigating the diagnostic accuracy of any test for PKDL. Study quality was assessed using QUADAS-2. Data were extracted from 21 articles including 43 separate studies. Twenty-seven studies evaluated serological tests (rK39 dipstick, ELISA, DAT, and leishmanin tests), six studies molecular tests, eight microscopy, and two cultures. Only a few of these studies reported a valid estimate of diagnostic accuracy, as most were case-control designs or used a reference standard with low sensitivity. The included studies were very heterogeneous, for example, due to a large variety of reference standards used. Hence, no summary estimates of sensitivity or specificity could be made. We recommend well-designed diagnostic accuracy trials that evaluate, side-by-side, all currently available diagnostics, including clinical symptoms, serological, antigen, molecular, and parasitological tests and possible use of statistical modelling to evaluate diagnostics when there is no suitable gold standard. PMID:23935641
Ma hmoudjanlou, H. (MSc
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.
Murari L Das
Full Text Available Prior to a community-based efficacy trial of long-lasting insecticidal nets (LLINs in the prevention of visceral leishmaniasis (VL; also called kala-azar, a pilot study on preference of tools was held in endemic areas of India and Nepal in September 2005.LLINs made of polyester and polyethylene were distributed to 60 participants, who used the nets sequentially for 7 d. Acceptability and preference were evaluated via indirect indicators through questionnaires at three defined time points before and after use of the LLINs and through focus group discussions (FGDs. In the latter, preferences for color and size were also assessed. Untreated bed nets were owned by 87% of the households prior to the study. All users liked textures of both LLIN types after 7 d of use, but had a slight preference for those made of polyester if they were to recommend a LLIN to relatives or friends (p<0.05, mainly because of their relatively greater softness in comparison to polyethylene LLINs. Users reported that both net types reduced mosquito bites and number of insects, including sand fly (bhusana; genus Phlebotomus, inside the house. Side effects were minor and disappeared quickly.The large-scale intervention trial considered the preferences of the study population to decide on the best tool of intervention--light-blue, rectangular, polyester LLINs of different sizes.
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.
Congwen Song; Manzhu Bao
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.
Marco-Medina, Ana; Casas, José Luis
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. PMID:23625080
Cassava is an important tuber crop grown in tropical and subtropical regions. Protocols for regeneration from tissue culture are very genotype dependent and in many cultivars quite ineffective, therefore improvement programmes through genetic engineering are restricted. Recently, protocols were developed for efficient somatic embryogenesis and/or organogenesis using zygotic embryos and nodal axillary meristems to minimize genotype dependency. Buds were regenerated directly from the exercised explants after a two step culture procedure. In the embryo explants, profilic shoot formation occurred within 2-3 weeks on a medium containing 0.5 mg I-1 BAP alone, or in combination with 0.1 mg I-1 NAA. Nodal explants with an axillary meristem were used to initiate a round, compact, 'bulb like' structure in the high cytokinin containing medium. In the presence of NAA (0.1 mg I-1), BAP (1 mg I-1) and GA3 (0.1 mg I-1), these structures produced multiple shoots. Efficient somatic embryogenesis was obtained from the cotyledon explants. Plants were obtained from all the African genotypes tested, although at different frequencies. The rooting of regenerated shoots exceeded 95% in phytohormone free medium. The polidy of the regenerants was examined using flow cytometry; no change in their ploidy level was observed. The random amplified polymorphic DAN (RAPD)-polymerase chain reaction (PCR) technique was employed to investigate molecular differences and also to generate polymorphic DNA markers in the in vitro regenerated Cassava plants. Twenty primers were used to generate patterns from several regenerants of two Cassava cultivars. The validity of using RAPD markers in such studies is discussed. (author). 3 refs, 1 tab
CHEN Liang; WANG Ping-sheng; Yamaguchi Satoshi
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.
Genetic variability of Brazilian isolates of Alternaria alternata detected by AFLP and RAPD techniques Variabilidade genética de isolados Brasileiros de Alternaria alternata por meio de marcadores moleculares de AFLP e RAPD
ógeno por meio da caracterização de vinte e quatro isolados de A. alternata f. sp. citri de plantas de citros juntamente com quatro isolados endofíticos de manga, sendo um deles identificado como Alternaria tenuissima e outros três como Alternaria arborescens. A análise de componentes principais (PCA do perfil de bandas obtidos pela aplicação de duas técnicas de marcadores moleculares, Amplificação Aleatória de Polimorfismos de DNA (RAPD e Polimorfismo de Comprimento de Fragmentos Amplificados (AFLP, mostrou a formação de quatro grupos distintos. Apesar do mais amplo perfil de análise por meio da técnica de AFLP, não foi observado nenhuma modificação significativa dentro dos grandes grupos obtidos quando comparado ao RAPD, exceto no posicionamento dos isolados LRS 39/3 e 25M. Além disso, em ambas as análises, somente os isolados de plantas de limão agruparam entre si. Considerando outros hospedeiros ou tecidos de planta não foi possível encontrar grupos específicos. Concluindo, ambas as análises (RAPD e AFLP são eficientes no estudo de variabilidade genética de Alternaria spp., fornecendo informações sobre a diversidade genética desta espécie, servindo como base para futuramente correlacionar este estudo com estudos adicionais objetivando o controle da doença.
ETOH, Takeomi; Watanabe, Hideki; Iwai, Sumio
RAPD variation of 30 garlic clones collected in the primary center of origin, Central Asia, was compared with that of 30 garlic clones collected in the westernmost area of distribution, the Iberian Peninsula. Central Asian garlic clones were complete-bolting type, and some of them were fertile clones. On the other hand, Iberian garlic clones showed incomplete-bolting type, and all of them were sterile clones. Basing on the genetic similarity, a dendrogram among those garlic clones by RAPD was...
Nagl Nevena; Taški-Ajduković Ksenija; Popović Andrea; Ćurčić Živko; Danojević Dario; Kovačev Lazar
In marker assisted breeding programs, determination of genome polymorphism and development of suitable molecular markers is of the greatest importance. The aim of this research was development of RAPD markers, which will enable quick and cost efficient DNA polymorphism analysis among closely related sugar beet genotypes. The research was conducted on twelve sugar beet genotypes from population of closely related genotypes. Reactions with eight RAPD primers ...
Impact assessment of contaminants in soil is an important issue in environmental quality study and remediation of contaminated land. A random amplified polymorphic DNA (RAPD) 'fingerprinting' technique was exhibited to detect genotoxin-induced DNA damage of plants from heavy metal contaminated soil. This study compared the effects occurring at molecular and population levels in barley seedlings exposed to cadmium (Cd) contamination in soil. Results indicate that reduction of root growth and increase of total soluble protein level in the root tips of barley seedlings occurred with the ascending Cd concentrations. For the RAPD analyses, nine 10-base pair (bp) random RAPD primers (decamers) with 60-70% GC content were found to produce unique polymorphic band patterns and subsequently were used to produce a total of 129 RAPD fragments of 144-2639 base pair in molecular size in the root tips of control seedlings. Results produced from nine primers indicate that the changes occurring in RAPD profiles of the root tips following Cd treatment included alterations in band intensity as well as gain or loss of bands compared with the control seedlings. New amplified fragments at molecular size from approximately 154 to 2245 bp appeared almost for 10, 20 and 40 mg L-1 Cd with 9 primers (one-four new polymerase chain reaction, (PCR) products), and the number of missing bands enhanced with the increasing Cd concentration for nine primers. These results suggest that genomic template stability reflecting changes in RAPD profiles were significantly affected and it compared favourably with the traditional indices such as growth and soluble protein level at the above Cd concentrations. The DNA polymorphisms detected by RAPD can be applied as a suitable biomarker assay for detection of the genotoxic effects of Cd stress in soil on plants. As a tool in risk assessment the RAPD assay can be used in characterisation of Cd hazard in soil
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
Full Text Available Yulianti, Siregar IZ, Wijayanto N, Tapa Darma IGK, Syamsuwida D (2011 Genetic variation of Melia azedarach in community forests of West Java assessed by RAPD. Biodiversitas 12: 64-69. Melia azedarach L. or mindi (local name is one of the widely planted exotic species in Indonesia, mostly found in community forests in West Java. However, improving and increasing the productivity of mindi commmunity plantation in West Java requires information on patterns of existing genetic diversity. The present work was aimed at estimating the genetic variation of mindi by using RAPD markers. Outcome of the activities was to propose appropriate conservation and management strategies of genetic resources in order to support the establishment of seed sources. Six populations of mindi plantation in the community forests were chosen for this research, i.e Sukaraja (Bogor-1, Megamendung (Bogor-2, Bandung, Purwakarta, Sumedang and Kuningan. Five primers (OPA-07, OPY-13, OPY-16, OPA-09 and OPO-05 producing reproducible bands were analysed for 120 selected mother trees in total, in which 20 trees per locality were sampled. Data were analysed using Popgene ver 1.31, NTSYS 2.02 and GenAlEx 6.3. Based on the analysis, the observed number of alleles per locus ranging from 1.43 to 1.60, and percentage of polymorphic loci (PPL ranging from 43.33 to 60.00.%. The levels of genetic variation were considered as moderate for all populations (He range from 0.1603 to 0.1956 and the the mean level of genetic diversity between population (Gst was 0.3005. Cluster analysis and Principal Coordinates showed three main groups, the first group consists of 4 populations i.e Bandung, Kuningan, Purwakarta and Megamendung, the second was Sukaraja and the third was Sumedang. Based on Analysis of Molecular Variance (AMOVA, the Percentages of Molecular Variance within population (69% is higher than that of between populations (31%. The moderate level of genetic variation in the community
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.
Random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) markers were used to study the DNA polymorphism in 12 gamma ray induced morphological mutants of blackgram [Vigna mungo (L.) Hepper]. Of the 35 random and eight ISSR primers used, 15 random and five ISSR primers detected polymorphism among the mutants. Total number of bands varied from 2 to 9 for RAPD and from 6 to 10 for ISSR. The number of polymorphic bands varied from 1 to 3 for RAPD and from 1 to 6 for ISSR. Percentage of polymorphism ranged from 12.5 to 50.0 for RAPD and 12.5 to 44.4 for ISSR. Significant DNA polymorphism among the mutants were observed using RAPD (25.8%) and ISSR (33.3%) marker. An young leaf chlorine mutant and a smooth pod mutant showed more DNA polymorphism as compared to the parent. DNA polymorphism data revealed by RAPD and ISSR could facilitate selection of mutants to be involved in cross breeding and genome mapping
Badaeva, E. D.; O. Yu. Shelukhina; Goryunova, S. V.; Loskutov, I. G.; V. A. Pukhalskiy
Tetraploid oat species Avena abyssinica, A. vaviloviana, A. barbata, and A. agadiriana were studied using C-banding technique, in situ hybridization with the 45S and 5S rDNA probes, and RAPD analysis in comparison with the diploid species carrying different types of the A-genome (A. wiestii, As; A. longiglumis, Al; A. canariensis, Ac; A. damascena, Ad, A. prostrata, Ap). The investigation confirmed that all four tetraploids belong to the same AB-genome group; however A. agadiriana occupies di...
ASHWANI KUMAR; NEELAM VERMA
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 avera...
Meera N., Lohithaswa HC, Niranjana Murthy and Shailaja Hittalmani
Genotype specific fingerprints were detected in two grain amaranthus varieties KBGA1 and Suvarna using SSR and RAPD markers. In this study 41 Pigeon Pea SSR markers and 6 RAPD markers were used to generate DNA fingerprints for the two varieties of grain amaranthus. Analysis of polymorphic fragments generated from SSR and RAPD markers revealed the genetic variation between grain amaranthus varieties KBGA1 and Suvarna. The results indicate that DNA markers are appropriate tools for assessing ge...
Wang, Pei; Lu, Yanli; Zheng, Mingmin; Rong, Tingzhao; Tang, Qilin
Genetic relationship of a newly discovered teosinte from Nicaragua, Zea nicaraguensis with waterlogging tolerance, was determined based on randomly amplified polymorphic DNA (RAPD) markers and the internal transcribed spacer (ITS) sequences of nuclear ribosomal DNA using 14 accessions from Zea species. RAPD analysis showed that a total of 5,303 fragments were produced by 136 random decamer primers, of which 84.86% bands were polymorphic. RAPD-based UPGMA analysis demonstrated that the genus Z...
Karine Cristina Bicalho
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
J. L. Domínguez Álvarez; R. Nieto-Ángel; A. F. Barrientos- Priego; J. P. Legaria-Solano; J. Pineda-Pineda
En la presente investigación se evaluó la variabilidad molecular y morfológica mediante marcadores tipo RAPDs (Polimorfismos en el ADN Amplificados al Azar), de plantas de guayabo (Psidium guajava L.) Media China derivadas de semilla y sometidas a baja disponibilidad de Fe y Zn. Se establecieron asociaciones entre diecisiete caracteres morfológicos y 49 loci RAPDs y la respuesta de las plantas al estrés nutrimental, mediante análisis de agrupamiento empleando el NTSYS, DARwin y #967;2. E...
C. Magarinos Torres
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
Gouri S. Bhunia
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.
Full Text Available INTRODUCTION: Post-kala-azar dermal leishmaniasis (PKDL is a cutaneous complication appearing after treatment of visceral leishmaniasis, and PKDL patients are considered infectious to sand flies and may therefore play a role in the transmission of VL. We estimated the risk and risk factors of PKDL in patients with past VL treatment in south-eastern Nepal. METHODS: Between February and May 2010 we traced all patients who had received VL treatment during 2000-2009 in five high-endemic districts and screened them for PKDL-like skin lesions. Suspected cases were referred to a tertiary care hospital for confirmation by parasitology (slit skin smear (SSS and/or histopathology. We calculated the risk of PKDL using Kaplan-Meier survival curves and exact logistic regression for risk factors. RESULTS: Out of 680 past-treated VL patients, 37(5.4% presented active skin lesions suspect of PKDL during the survey. Thirty-three of them underwent dermatological assessment, and 16 (2.4% were ascertained as probable (2 or confirmed (14 PKDL. Survival analysis showed a 1.4% risk of PKDL within 2 years of VL treatment. All 16 had been previously treated with sodium stibogluconate (SSG for their VL. In 5, treatment had not been completed (≤ 21 injections. Skin lesions developed after a median time interval of 23 months [interquartile range (IQR 16-40]. We found a higher PKDL rate (29.4% in those inadequately treated compared to those who received a full SSG course (2.0%. In the logistic regression model, unsupervised treatment [odds ratio (OR = 8.58, 95% CI 1.21-374.77], and inadequate SSG treatment for VL in the past (OR = 11.68, 95% CI 2.71-45.47 were significantly associated with PKDL. CONCLUSION: The occurrence of PKDL after VL treatment in Nepal is low compared to neighboring countries. Supervised and adequate treatment of VL seems essential to reduce the risk of PKDL development and active surveillance for PKDL is needed.
Ana Cruz Morillo Coronado; Liseth Gómez Beltrán; Iván Ávila Morales; Ernesto Andrade Urresta; Yacenia Morillo Coronado
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 Vitamin...
Puy, D.; Chimenos Küstner, Eduardo; C. Dorado
De entre las lesiones observadas en la cavidad bucal, las producidas por el virus Herpes simplex (VHS) y las originadas en la estomatitis aftosa recidivante (EAR) representan una parte importante de las lesiones que el odontólogo o estomatólogo encuentra cotidianamente. Aunque cada uno de estos dos procesos posee una etiopatogenia y unas características diferentes, en algunas ocasiones su diagnóstico puede llegar a ser controvertido y difícil, debido a su similar apariencia clínica, planteánd...
Estrella, María Soledad
La conferencia aborda la enseñanza de la estadística, destacando tres modelos específicos del área: la guía GAISE, el ciclo investigativo PPDAC, y el ambiente para el aprendizaje del razonamiento estadístico SRLE. Además, se centra en el desarrollo del pensamiento estadístico según la jerarquía cognitiva de alfabetización estadística, razonamiento y pensamiento estadístico.
This study was performed to assess chemical components and genetic variability of five Syrian wheat varieties using NIR, RAPD and AFLP techniques. NIR technique showed that Cham6 was the best variety in term of wheat grain quality due to their lowest protein (%), hardness, water uptake and baking volume and the highest starch (%) compared to the other tested varieties. PCR amplifications with 21 RAPD primers and 13 AFLP PCs primer combinations gave 104 and 466 discernible loci of which 24 (18.823%) and 199 (45.527%) were polymorphic for the both techniques respectively. Our data indicated that the three techniques gave similar results regarding the degree of relatedness among the tested varieties. In the present investigation, AFLP fingerprinting was more efficient than the RAPD assay. Where the letter exhibited lower Marker Index (MI) average (0.219) compared to AFLP one (3.203). The pattern generated by RAPD, AFLPs markers or by NIR separated the five wheat varieties into two groups. The first group consists of two subclusters. The first subcluster involved Cham8 and Bohous6, while the second one includes Cham6 that is very closed to precedent varieties. The second group consists of Bohous9 and Cham7 that were also closely related. Based on this study, the use of NIR, RAPD and AFLP techniques could be a powerful tool to detect the effectiveness relationships of these technologies. (author)
YUYU SURYASARI POERBA
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.
Casiva, Paola V; Saidman, Beatriz O; Vilardi, Juan C; Cialdella, Ana M
Morphological and genetic diversity among Acacia aroma, A. macracantha, A. caven, and A. furcatispina were studied with morphometric, isozymal, and RAPD approaches. The analysis of seven isozyme systems revealed 21 loci, and RAPD analysis showed 34 loci. Most of these loci allowed us to differentiate the species, with the exception of A. aroma and A. macracantha, the two most similar species. The levels of genetic variability estimated by isozymes were higher than those obtained from RAPD analyses. Morphometric characters showed highly significant differences among the species, although A. aroma and A. macracantha are differentiated only by thorn length. The phenogram obtained from isozyme data is consistent with morphological data. The RAPD phenogram based on allelic frequencies showed agreement with morphological and isozymal approaches only at the intraspecific levels, while the RAPD phenogram based on Nei and Li's similarity measures agreed with the phenograms constructed from isozyme and morphological data. High similarities and high indirect gene flow were found between A. aroma and A. macracantha, results that call the relationship between them into question. PMID:21665685
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.
Full Text Available Randomly Amplified Polymorphic DNA (RAPD is a novel procedure for the identification of polymorphism in plants based on PCR. It does not require prior knowledge of a DNA sequence. RAPD markers are based on the amplification of unknown DNA sequences using single, short, random oligonucleotide primers. Many of the technical limitations of RFLPs have been overcome by RAPD. In the present study seven varieties of C. annuum L. were analysed for RAPD polymorphism using 5 random primers. The study shows that the varieties show remarkable genetic variation. RAPD primers showed different DNA fingerprints for different varieties of Capsicum L. studied. Variation was observed at varietal level in C. annuum L. with respect to the morphological traits, phytoconstituents estimated ( capsaicin, sugar and vitamin C and karyotype analysis also. Thus all the seven can be distinguished at varietal level. Therefore they can be used as a suitable source material for future breeding, genetic and other experiments. The use of other molecular markers like SSRs, AFLP, etc. would provide more precise estimates of genetic variability in the varieties.
Boerhavia diffusa is a medicinally important plant and finds extensive uses in traditional herbal drug preparations. For the development of improved varieties in terms of superior yield and quality of herb/root of B. diffusa, mutation breeding was attempted. Mutants generated by physical and chemical mutagenic treatments were screened for yield and quality parameters of the root/herb up to three consecutive generations. The selected-screened lines generated by physical and chemical mutagenic treatments on two selected genotypes I and II were molecularly analyzed using eight isozymes and eleven RAPD primers producing good amplification. Mutants from BD10 (selected genotype I) were distinct, while, in case of BD22 (selected genotype II), only one mutant BDMu7 was recorded distinct by isozyme analysis. The wild mutant (BDMu16, with maximum height and mouve coloured flower) was distinct in RAPD banding pattern. Isozymes differentiated the mutants from their respective controls, whereas RAPD differentiated the mutants and controls and also distinguished the mutants. The RAPD analysis was found to be better suited than isozymes for detecting genetic differences among controls and their mutants. However, both RAPD and isozyme analyses gave similar patterns of genetic relationships
Esfera pública y juegos de azar: del meeting contra el juego al allanamiento de domicilio privado. Prensa, parlamento y policía en Buenos Aires (1901-1902 Esfera Pública e jogos: do meeting contra o jogo ao allanamiento de domicílio privado. Imprensa, parlamento e polícia em Buenos Aires, (1901-1902 Public Sphere and Gambling: from the meeting against Gambling to Legal Home Search. Press, Parliament and Police in Buenos Aires, (1901-1902
Ana Victoria Cecchi
Full Text Available El 29 de septiembre de 1901 se realizó, en la ciudad de Buenos Aires, un meeting contra el juego concurriendo a la Plaza de Mayo unas 5.000 personas que solicitaron del Presidente de la República la represión de los juegos de azar en el ámbito urbano. El meeting contra el juego permite indagar el lugar ocupado por la prensa en la organización de una movilización de estas características, cuyo eje de protesta encuentra sus fundamentos en el terreno moral y pone en evidencia que no todas las iniciativas nacidas en el seno de la opinión pública van en el sentido de la ampliación de derechos. Las discusiones parlamentarias que esta movilización proyecta también resultan de interés en la medida en que derivan, en 1902, en la sanción de la ley de Represión del juego que volverá legal el allanamiento de domicilio privado.Em 29 de Setembro de 1901 realizou-se na cidade de Buenos Aires um meeting contra o jogo, participando 5.000 pessoas na Plaza de Mayo que solicitaram ao Presidente da República a repressão dos jogos de azar nas áreas urbanas. O meeting contra o jogo permite examinar o lugar da imprensa na organização de uma mobilização destas características, cujo eixo de protesto encontra seus fundamentos no território moral e evidencia que nem todas as iniciativas surgidas na opinião pública refletem a ampliação de direitos. Os debates parlamentários que esta mobilização suscitou também interessam porque -depois de varios meses- derivam, em 1902, na sanção de uma Lei de repressão ao jogo que tornará legal a invasão policial de domicílio particular.On September the 29th, 1901, a meeting against gambling was held in Buenos Aires: about 5,000 people attended the Plaza de Mayo and demanded that the President of the Republic repressed games of chance in urban environments. This meeting against gambling provides grounds for an inquiry regarding the role the media plays in the organization of such events, which start
Full Text Available Olive (Olea europea L. is one of the oldest cultivated plants characteristic in the Mediterranean area, where it is the most important oilproducing crop. The cultivated olive (O. europaea L. var. europaea is propagated by cutting or grafting, whereas wild olive (O. europaea L. var. sylvestris is reproduced from seeds. These two olive types are interfertile and have led to a large number of varieties. Morphological descriptions are not entirely reliable, due to numerous synonyms and homonyms in designations, labelling mistakes, the presence of varietal clones, and the uncertain identification methods thus far applied. Molecular markers, as random amplified polymorphic DNA (RAPD markers, are environment-independent and efficient to identify olive varieties and to detect synonymous and homonymous. In this study, fifteen selected RAPD markers are used for determination of relationships among twenty individuals belonging to four important Turkish olive cultivars. Our results showed that RAPD markers can be used to differentiate olive cultivars
Xiao-lin YIN; Kai-tai FANG; Yi-zeng LIANG; Ricky NS WONG; Amber WY HA
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.
Wilkerson, R C; Parsons, T J; Albright, D G; Klein, T A; Braun, M J
The usefulness of random amplified polymorphic DNA (RAPD) was examined as a potential tool to differentiate cryptic mosquito species. It proved to be a quick, effective means of finding genetic markers to separate two laboratory populations of morphologically indistinguishable African malaria vectors, Anopheles gambiae and An. arabiensis. In an initial screening of fifty-seven RAPD primers, 377 bands were produced, 295 of which differed between the two species. Based on criteria of interpretability, simplicity and reproducibility, thirteen primers were chosen for further screening using DNA from thirty individuals of each species. Seven primers produced diagnostic bands, five of which are described here. Some problematic characteristics of RAPD banding patterns are discussed and approaches to overcome these are suggested. PMID:8269099
Ali, A. M.
Full Text Available The routine identification of mycobacterial strains isolated from patients in different locations in Egypt was confirmed by specific DNA fragment amplification. The susceptibilities of 72 Mycobacterium tuberculosis strains against the four antibiotics used in tuberculosis treatment (Isoniazid, INH; Rifampicin, Rif; Streptomycin, St and Ethambutol, E were examined. Our results indicated that, multi drug resistant tuberculosis (MDR-TB represents about 19.5% of the tested strains, whereas sensitive strains represented 26.4%. The genetic polymorphism of the tested strains was examined using RAPD analysis. Six selected strains represent the different antibiotic susceptibility groups were examined using RAPD fingerprinting. No difference between the strains was recorded using the RFLP analysis of amplified specific fragment. The discrimination power of RAPD analysis was inadequate to clarify the genetic correlation between the tested strains. MDR-TB was approximately double time in 2008 compared with the value in 2007. Most of the new MDRTB was correlated with resident dense population regions.
Full Text Available RAPD markers were used to investigate the genetic diversity of 31 Brazilian cassava clones. The results were compared with the genetic diversity revealed by botanical descriptors. Both sets of variates revealed identical relationships among the cultivars. Multivariate analysis of genetic similarities placed genotypes destinated for consumption "in nature" in one group, and cultivars useful for flour production in another. Brazil?s abundance of landraces presents a broad dispersion and is consequently an important resource of genetic variability. The botanical descriptors were not able to differentiate thirteen pairs of cultivars compared two-by-two, while only one was not differentiated by RAPD markers. These results showed the power of RAPD markers over botanical descriptors in studying genetic diversity, identifying duplicates, as well as validating, or improving a core collection. The latter is particularly important in this vegetatively propagated crop.
Rafael Gustavo Ferreira Morales
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.
Leonardo do Nascimento Rolim
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.
Hazarika, Dimendra; Thangaraj, M; Sahu, Sunil Kumar; Kathiresan, K
Genetic diversity was analysed in three populations of the mangrove species, Avicennia marina by using random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR). Ten random decamer primers were used to score the diversity from three locations of eastcoast of India: Parangipettai (Tamil Nadu), Kakkinada (Andhra Pradesh) and Sundarbans (West Bengal). These primers produced 388 scorable DNA fragments, of which 252 (64.98%) were polymorphic, 182 (46.90%) were monomorphic, and 14 (3.61%) were unique. RAPD banding patterns displayed variations between and within the populations, while, there was no morphological variation. PMID:24617156
SongShujuan; Robert.C.ClarkeI; ShaoHong
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.
Irshad, Saba; Singh, Jyotsna; Kakkar, Poonam; Mehrotra, Shanta
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. PMID:19100816
Andréa Alves do Egito; Beatriz Helena Fuck; Concepta McManus; Samuel Rezende Paiva; Maria do Socorro Maués Albuquerque; Sandra Aparecida Santos; Urbano Gomes Pinto de Abreu; Joaquim Augusto da Silva; Fabiana Tavares Pires de Souza Sereno; Arthur da Silva Mariante
Blood samples were collected from Pantaneiro Horses in five regions of Mato Grosso do Sul and Mato Grosso States. Arabian, Mangalarga Marchador and Thoroughbred were also included to estimate genetic distances and the existing variability among and within these breeds by RAPD-PCR (Random Amplified Polymorphic DNA - Polymerase Chain Reaction) molecular markers. From 146 primers, 13 were chosen for amplification and 44 polymorphic bands were generated. The analysis of molecular variance (AMOVA)...
Sa'ad, Tata Umar; Sabo, Shehu; Abdullahi, Aliyu Dahuwa
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…
Manna, P P; Bharadwaj, D.; Bhattacharya, S.; Chakrabarti, G; Basu, D; Mallik, K K; Bandyopadhyay, S.
Indian kala-azar patients have normal numbers of peripheral blood NK cells but impaired functional activity due to decreased binding and lysis of target cells. This impairment of NK activity could not be corrected by exogenous recombinant human alpha or gamma interferon. However, recombinant human interleukin 2 was able to restore this activity by augmenting conjugate formation and lysis of target cells.
Theinert, Stephan M.; Basu, Rajatava; Forgber, Michael; Roy, Syamal; Sundar, Shyam; Walden, Peter
Sera of kala-azar patients from Bihar, India, were used to identify Leishmania donovani antigens encoded by a phage expression library. Ten antigens were identified, five of which have not been described as leishmania antigens before. The antigens specifically react with sera of leishmania-infected patients but not of toxoplasma- or plasmodium-infected patients.
Ismail, A; Kharazmi, A; Permin, H;
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...
E. D. Badaeva
Full Text Available Tetraploid oat species Avena abyssinica, A. vaviloviana, A. barbata, and A. agadiriana were studied using C-banding technique, in situ hybridization with the 45S and 5S rDNA probes, and RAPD analysis in comparison with the diploid species carrying different types of the A-genome (A. wiestii, As; A. longiglumis, Al; A. canariensis, Ac; A. damascena, Ad, A. prostrata, Ap. The investigation confirmed that all four tetraploids belong to the same AB-genome group; however A. agadiriana occupies distinct position among others. The C-banding, FISH, and RAPD analyses showed that Avena abyssinica, A. vaviloviana, and A. barbata are very similar; most probably they originated from a common tetraploid ancestor as a result of minor translocations and alterations of C-banding polymorphism system. AB-genome species are closely related with the A-genome diploids, and an As-genome species may be regarded as the most probable donor of their A-genome. Although their second diploid progenitor has not been identified, it seems unlikely that it belongs to the As-genome group. The exact diploid progenitors of A. agadiriana have not been determined; however our results suggest that at least one of them could be related to A. damascena.
ARIFIN NOOR SUGIHARTO
Full Text Available RAPD-PCR method and isozyme analysis were used to obtain information of genetic relationship among cucumber varieties. Such information is urgently utilized to support plant breeding program of cucumber. Research was done at the Biotechnology Laboratory of Plant Pest and Diseases, Faculty of Agriculture of Brawijaya University, Malang and Molecular Biology Laboratory, Faculty of Matemathic and Natural Sciences of Brawijaya University, Malang. DNA isolation was done using CTAB method by additional NaCl modification. Sixteen primers from operon were employed to amplify DNA genome by RAPD-PCR. Two enzymes, Esterase and AAT were chosen for isozyme analysis. Clad 97 Program was used for analyzing the data and results in data grouping based on proximity value. Cluster analysis based on isozyme data indicated that there was an adequate lower genetic variation in cucumber, where seven of nine tested varieties showed proximity value of 1.00. Eleven of sixteen primers in RAPD-PCR analysis produced DNA bands. Relativity analysis by using RAPD-PCR method showed high enough of genetic variation. Relativity analysis by using both methods showed that variety 07 was the furthermost. The proximity value between varieties 01 and 02 was 0.916667, these varieties have the higest proximity value among all varieties.
Bianca W. Bertoni
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.
Privitera, Claudio M; Stark, Lawrence W
The human pupil is an important element studied in many clinical procedures. The binocular pupil model presented has a topology encompassing much of the complexity of the pupil system neurophysiology. The dynamic parameters of the model were matched against pupil experiments under multiple conditions. It simulates responses to the swinging flashlight test for different degrees of relative afferent pupil defects, RAPD. PMID:17271776
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.
Information on genetic diversity and relationships among rice genotypes from Pakistan is currently very limited. Molecular marker analysis can truly be beneficial in analyzing the diversity of rice germplasm providing useful information to broaden the genetic base of modern rice cultivars. The objective of this study was to evaluate the genetic polymorphism of 75 rice accessions and improved cultivars using random amplified polymorphic DNA (RAPD) technique. Twenty-eight decamer-primers generated a total of 145 RAPD fragments, of which 116 (80%) were polymorphic. The number of amplification products produced by each primer varied from 3 to 9 with an average of 5.2 alleles primer-1. The size of amplified fragments ranged from 250 to 4000bp. A dendrogram was generated from minimal variance algorithm using Ward method. All the 75 genotypes were grouped into two main groups corresponding to aromatic and non-aromatic types of indica rice. Clustering of accessions did not show any significant pattern of association between the RAPD fingerprints and collection sites. This type of analysis grouping different rice accessions in relation to fragrance, a major rice quality determinant, and varietal group is extremely useful to develop a core collection and gene bank management. Further more, the information revealed by the RAPDs regarding genetic variation is helpful to the plant breeder in selecting diverse parents and for future orientation of rice breeding program. (author)
Bruno Guedes Alcoforado Aguiar
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.
Konstanze Ursula Behrmann
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.
Wang, Tiegu; Huang, Qunce; Feng, Weisen
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, s1, opt-16, and f14, were significantly associated with the muticate trait, while three SSR markers, Rht8 (Xgwm261), Rht-B1b, and Rht-D1b, 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.
Mamta Gupta; Bhawna Verma; Naresh Kumar; Rakesh K. Chahota; Rajeev Rathour; Shyam K. Sharma; Sabhyata Bhatia; Tilak R. Sharma
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.
Jiang, Y; Liu, J-P
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. PMID:22179965
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, s1, opt-16, and f14, were significantly associated with the muticate trait, while three SSR markers, Rht8 (Xgwm261), Rht-B1b, and Rht-D1b, 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
Salah E.D. El-Assal
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
Genetic variation among South Brazilian accessions of Lippia alba Mill. (Verbenaceae) detected by ISSR and RAPD markers Variabilidade genética entre acessos sul-brasileiros de Lippia alba Mill. (Verbenaceae) detectada por ISSR e RAPD
MF. Manica-Cattani; J. Zacaria; G Pauletti; L. Atti-Serafini; S Echeverrigaray
Twenty-seven accessions of Lippia alba Mill. collected in Rio Grande do Sul state, Brazil, were analysed by ISSR and RAPD markers to evaluate their genetic variability and relationships. Six ISSR primers and four RAPD primers generated 120 amplified fragments, most of which were polymorphics. The overall genetic variability among accessions was very high when compared with other plant species. The hierarchical analysis of molecular data (UPGMA) showed low relationship between accessions, and ...
Random amplified polymorphic DNA (RAPD) markers were successfully used in discrimination of sexes in Nile tilapia fish ( Oreochromis niloticus) using linear discriminant function analysis. The results provide support for the view that major genetical sex determining factors exist in tilapia.
Paula Menna Barreto Dias
Full Text Available The genetic diversity in the red clover USA core collection was measured by RAPD and isozyme markers,comparing different approaches of isozyme data analysis and comparing the results of RAPD with those of isozymes. A totalof 15 isozyme loci and 114 RAPD fragments were analyzed. Genetic diversity measured with RAPD and isozymes was high.The analyses of isozymes as binary data was highly correlated with the allele-frequency approach. There was no correlationbetween the distance matrices of different markers. Isozyme data grouped the accessions of red clover in four groups. Thevariation evidenced by AMOVA was higher among and within groups in the populations than at the intra-population level.Results showed the high diversity in the red clover core collection and indicated some populations that could be used inbreeding programs of the crop in Brazil.
Lund, Flemming; Nielsen, A.B.; Skouboe, P.
using morphotypes (colony morphology and colours) and secondary metabolite profiles. Based on production of secondary metabolites the P. commune isolates were classified into 6 groups. The genetic diversity of the P. commune isolates was assessed using randomly amplified polymorphic DNA (RAPD......) and amplified fragment length polymorphism, (AFLP). For a sub-set of 272 P. commune isolates RAPD analysis generated 33 RAPD groups whereas AFLP profiling revealed 55 AFLP groups. This study conclusively showed that the discriminatory power of AFLP was high compared to RAPD and that AFLP fingerprinting matched......In an 8-year study of the diversity and distribution of Penicillium commune contaminants in two different cheese dairies, swab and air samples were taken from the production plants, the processing environment and contaminated cheeses. A total of 321 Penicillium commune isolates were characterized...
Berić Tanja; Urdaci Maria C.; Stanković S.; Knežević-Vukčević Jelena
Genetic diversity and production of hydrolytic enzymes of 205 Bacillus isolates from different geographical and ecological niches in Serbia were studied. Combining RAPD analysis and 16S DNA sequencing, we determined 13 different groups of RAPD profiles within four (five) species: B. subtilis, B. cereus/B. thuringiensis, B. pumilus, and B. firmus. Screening for production of hydrolytic enzymes showed that there was no correlation of enzyme production with species. Most of the isolates from all...
DINESH CHANDRA; PREETI CHOUDHARY
Under a survey and collection programme of National Agricultural Technology Project (NATP), various accessions of Aloe vera were collected from Rajasthan and Gujarat. In this study 10 Accessions from Rajasthan were characterized through RAPD. RAPD revealed 32.08 per cent polymorphic bands detecting 11.9% average diversity among the accessions studied. The diversity ranged from 4.3% to 20.4%. The primers like OPG-15 having high PIC value (0.346) are considered important for diversi...
Elisabete Yurie Sataque Ono
Full Text Available In this study a total of 16 Fusarium verticillioides strains isolated from corn feed samples were characterized by fumonisin (FB production and random amplified polymorphic DNA (RAPD. All the strains produced FB1 and FB2 with levels ranging from 2.41 to 3996.36 µg/g, and from 1.18 to 1209.91 µg/g, respectively. From the 16 F. verticillioides strains, four were identified as low (3.59 to 1289.84 µg/g, eight as intermediate (>1289.84 to 3772.44 µg/g and four strains as high (>3772.44 µg/g fumonisin producers. From the total of 105 loci amplified, 60 (57.14% were polymorphic. RAPD analysis showed very similar patterns among low, moderate and high fumonisin-producing strains. Although RAPD markers were capable of discriminating the different F. verticillioides strains, there was no clear association between these makers and fumonisin production.Neste estudo, 16 cepas de F. verticillioides isoladas de amostras de ração de milho foram caracterizadas com base na produção de fumonisinas (FB e em marcadores de polimorfismos de DNA amplificado ao acaso (RAPD. Todas as cepas produziram FB1 e FB2, com níveis variando, respectivamente, de 2,41 a 3996,36 µg/g e 1,18 a 1209,91 µg/g. De acordo com a produção de fumonisinas totais (FB1 + FB2 e a distribuição por análise de quartis, do total de 16 cepas de F. verticillioides, quatro foram identificadas como baixas produtoras de fumonisinas (3,59 a 1289,84 µg/g, oito como intermediárias (>1289,84 a 3772,44 µg/g e quatro como altas produtoras de fumonisinas (>3772,44 µg/g. Os 10 primers utilizados amplificaram 105 locos, 60 (57,14% dos quais foram polimórficos. As análises de RAPD mostraram padrões muito similares entre as cepas baixas, médias e altas produtoras de fumonisinas. Embora os marcadores RAPD tenham se mostrado capazes de discriminar as diferentes cepas de F. verticillioides, não foi detectada nenhuma associação entre estes marcadores e a produção de fumonisinas.
Minu Sherry; Steve Smith; Ashok Patel; Phil Harris; Paul Hand; Liz Trenchard; Janey Henderson
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.
Full Text Available Abstract Background Rapid, easy, economical and accurate species identification of yeasts isolated from clinical samples remains an important challenge for routine microbiological laboratories, because susceptibility to antifungal agents, probability to develop resistance and ability to cause disease vary in different species. To overcome the drawbacks of the currently available techniques we have recently proposed an innovative approach to yeast species identification based on RAPD genotyping and termed McRAPD (Melting curve of RAPD. Here we have evaluated its performance on a broader spectrum of clinically relevant yeast species and also examined the potential of automated and semi-automated interpretation of McRAPD data for yeast species identification. Results A simple fully automated algorithm based on normalized melting data identified 80% of the isolates correctly. When this algorithm was supplemented by semi-automated matching of decisive peaks in first derivative plots, 87% of the isolates were identified correctly. However, a computer-aided visual matching of derivative plots showed the best performance with average 98.3% of the accurately identified isolates, almost matching the 99.4% performance of traditional RAPD fingerprinting. Conclusion Since McRAPD technique omits gel electrophoresis and can be performed in a rapid, economical and convenient way, we believe that it can find its place in routine identification of medically important yeasts in advanced diagnostic laboratories that are able to adopt this technique. It can also serve as a broad-range high-throughput technique for epidemiological surveillance.
Gustavo Ribeiro Xavier
Full Text Available O conhecimento da variabilidade genética e da relação entre diferentes acessos de caupi é importante para maximizar o uso dos recursos genéticos disponíveis. Esse trabalho teve como objetivo avaliar a variabilidade genética em 45 acessos de caupi (Vigna unguiculata (L. Walp. oriundos do Brasil, EUA e Nigéria, por meio de marcadores RAPD. Foram encontrados 8 iniciadores polimórficos e um total de 48 bandas informativas. De acordo com os perfis polimórficos obtidos, foi observada a formação de quatro grupos genotípicos. Houve uma tendência de agrupamento em razão da origem dos acessos. A maioria dos acessos de variedades locais brasileiras pertence apenas a um grupo, o que sugere uma limitação da base genética. Vale ressaltar que nesse grupo não estavam presentes acessos da Nigéria considerados portadores de características agronômicas superiores, como, por exemplo, alta produtividade. RAPD é uma ferramenta eficiente, capaz de auxiliar a seleção de genótipos de caupi adaptados às diferentes condições edafo-climáticas brasileiras, com vistas ao aumento da produtividade e melhoria de outras características que atendam aos interesses regionais específicos.The knowledge on genetic variability and the relationship among different cowpea accesses is important to maximize resource use represented by available cowpea genotypes. The objective of this work was to determine the genetic variability among 45 cowpea (Vigna unguiculata (L. Walp. accesses from Brazil, USA and Niger, characterized by RAPD (Random Amplified Polymorphic DNA markers. Eight polymorphic primers were identified, comprehending a total of 48 informative bands. Based on the obtained polymorphic profiles, four major clusters were formed. Clustering was mainly influenced by the genotype origin. Most accesses from Brazilian landraces belong to just one cluster, suggesting a limited genetic basis. It is worth noting that none of the genotypes from Niger
Perry, Patricia; Mesa, Vilma María; Fernández, Felipe; Gómez, Pedro
Este libro va dirigido, principalmente a estudiantes de Ciencias Sociales, quienes necesitan conocer y manejar conceptos elementales de estadística. La presentación de los temas se hace a partir de situaciones concretas que motivan y justifican su importancia. Para llegar a expresar la forma operativa de los conceptos se desarrolla un proceso de búsqueda, con énfasis experimental, con el que se pretende aclarar paulatinamente, el concepto y además proporcionar al estudiante un modelo de una m...
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.
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.
Signal grass (Brachiaria decumbens) has been used for many years as grazing pasture for ruminants but it may know to be toxic to small ruminants. Mutagenesis provides an alternative approach to create phenotypic variability in the grass that might be free from toxicity and allow greater selection potential for desired characteristics. The mutant of Brachiara decumbens, which was obtained through induced mutation by gamma irradiation at dose 900 Gh showed phenotypic changes in term of tiller number, leaf to stem ratio and internode length. The variations caused by irradiation were detected by Random Amplified Polymorphic DNA (RAPD) analysis. Decamer oligonucleotide primers were used to generate DNA profiles. 575 bp polymorphic DNA band was observed between control and the mutant. RAPD has been proven to be useful to detect polymorphism between mutants and the control. (Author)
Wang Chao; Li Xin; Gao Li-na; Zhang Lu; Liu Wei; Liu Hui-min; Chen Ya-jun
A total of 69 random primers were screened by using random amplified polymorphic DNA (RAPD) markers to analyze the genetic bands of 32 Kentucky bluegrass cultivars. A total of 197 bands were amplified from 46 primers, among which 195 bands were polymorphic. Each primer could amplify one to nine polymorphic bands with an average of 4.3 per primer. Based on similarity coefficient analysis of RAPD results and by using NTSYS software to cluster analyze with the average UPGMA method, the result showed that 18 cultivars of the 32 were in group 1, three cultivars were in group 2, two cultivars were in group 3, eight cultivars were in group 4, and only one cultivar in group 5.
SUN Jun-ming; WU Shu-ming; TAO Wen-jing; DING An-lin; HAN Fen-xia; JIA Shi-rong
Near-isogenic lines of soybean lipoxygenase, which contain genes Lox, lx1, lx2, lx3, lx1.3, lx2.3,respectively, were used for polymorphic analysis by RAPD technique.520 10-mer-oligonucleotide primers were screened, and thirteen primers showed polymorphism among near-isogenic lines.There were six primers showed special polymorphic bands among lines lx1 and lx1.3. Especially,primer S352 presented the stable results in which a 900 bp band was found in the lines lx1 and lx1.3, and primer S352900 was detected with F2 generation of cross 96P11×Century-1, indicated primer S352900 could be identified as a RAPD marker linked to gene lx1 in soybeans, the distance of linkage was 7.6 cM.
Dimopoulos, G; Zheng, L; Kumar, V; della Torre, A; Kafatos, F C; Louis, C
Randomly amplified polymorphic DNA (RAPD) markers have been integrated in the genetic and cytogenetic maps of the malaria vector mosquito, Anopheles gambiae. Fifteen of these markers were mapped by recombination, relative to microsatellite markers that had been mapped previously. Thirty-four gel-purified RAPD bands were cloned and sequenced, generating sequence tagged sites (STSs) that can be used as entry points to the A. gambiae genome. Thirty one of these STSs were localized on nurse cell polytene chromosomes through their unique hybridization signal in in situ hybridization experiments. Five STSs map close to the breakpoints of polymorphic inversions, which are notable features of the Anopheles genome. The usefulness and limitations of this integrated mosquito map are discussed. PMID:8725241
Shioda, Hiroko; Satoh, Kanako; Nagai, Fumiko; Okubo, Tomoko; Seto, Takako; Hamano, Tomoko; Kamimura, Hisashi; Kano, Itsu
Juice and integument of leaves of 3 Aloe species, Aloe vera, A. ferox and A. africana, are not allowed to be used as food according to the Pharmaceutical Affairs Law in Japan. On the other hand, whole leaves of A. arborescens can be used as food. The present study was designed to distinguish Aloe species by random amplified polymorphic DNA (RAPD) analysis. DNA was isolated from fresh and dried leaves of the 4 Aloe species. Five out of 32 different 10-mer primers examined were useful for analysis. By comparison of the characteristic bands of PCR products on agarose gel, it was possible to distinguish the 4 species. Thus, the botanical species of Aloe in commercial food products can be identified by RAPD analysis. PMID:14606430
Imtiaz Ahmad Khan
Full Text Available Development of genetically compensating nullisomic-tetrasomic and ditelosomic lines of commonwheat (Triticum aestivum L. have been widely used to construct high density genetic maps of homoeologouswheat chromosomes. During present research, easier, cheaper and quicker procedure of Polymerase ChainReaction (PCR was used to map Randomly Amplified Polymorphic DNA primers on chromosome 1D ofcommon wheat. Genomic DNA was isolated from two genetic stocks of wheat cultivar Chinese Spring viz;NT-1D1B and NT-2A2B. PCR were conducted using RAPD primers GLC-07 and GLC-11. RAPD primerGLC-11 amplified a polymorphic allele of approximately 500 bp, which was present in NT-2A2B (used aspositive control but was absent in NT-1D1B indicating that the locus is present on chromosome 1D of commonwheat. Hence this marker (GLC-11 can reliably be used to keep track of chromosome 1D of hexaploid wheat.
KOSHY E. P.
Full Text Available The molecular marker based methods are highly preferred over other methods to assess the clonal fidelity andgenetic polymorphism. In this study, Random amplified polymorphic DNA (RAPD markers were used toamplify DNA from invitro propagated plantlets ofPsophocarpus tetragonolobus (L. DC (Winged Bean forevaluating the genetic stability. Seventeen arbitrary decamer primerswere used to find out the RAPD patternsexisting in parent plant and in vitroregenerated plantlets. These 17 primers (C61-C77 yielded 170 products ofwhich 44 were polymorphic (that is, 25.88% polymorphism.The dendrogram based on the Unweighted PairGroup Methodwith Arithmetic average (UPGMA cluster analysis and Nei’s similarity index depicted that theparent plant and the in vitro cultured plants have high degree of similarity
Full Text Available Cuscuta reflexa (Convolvulaceae is a holoparasitic vine that attacks the aerial parts of many shrubs, trees, and is used immensely in the Indian system of medicine. In this study, the randomly amplified polymorphic DNA (RAPD technique was employed for authentication of Cuscuta reflexa and its adulterant Cuscuta chinensis. Thirty two decamer oligonucleotide primers were used to amplify the genomic DNA isolated from the dried stems as well as seeds of both the species. Out of the thirty two primers used, fourteen did not amplify, eleven gave faint and non-reproducible, while seven gave species-specific reproducible unique bands. The unique bands obtained in PCR amplification clearly discriminated the two species, having similar morphology and thus, RAPD may serve as a complementary tool for quality control.
HU Guofu; YUAN Qiang; LI Fenglan; WEI Qi; HU Baozhong
In this study, we used RAPD to analyze four kinds of color-flowered Salvia splendens Ker-Gawl, and the optimal RAPD reaction conditions were the optimal reaction mixture (25 uL total volume) that contained 2.0 μL 10×buffer,0.45 mmol·L-1 dNTPs, 2.0 mmol·L-1 Mg2+, 2 U Taq DNA polymerase, 0.30 umol·L-1 primer and 40 ng genomic DNA. Total 84 bands were amplified from 12 primers used, and the differential bands had 28 bands, which was 33% of total bands. In cluster group analysis, the four kinds of color-flowered were divided into two styles. One style is that the red color and red-white color were grouped together, then they grouped with purple color into one cluster, and the white color was another style.
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.
Khyati D. Bhatt, Smita K. Girnari, Viralkumar B. Mandaliya, Lalit D. Chariya, Vrinda S. Thaker
Full Text Available Neem (Azadirachta indica A. Juss. is commercially valuable medicinal plant studied for molecular analysis. In the present study, a morphological variant of neem was observed near our department. The leaf pattern was abnormal (crimpled or curly morphologically observed and was against normal leaves. This curly leaves containing neem tree considered as mutant against normal. It was considered as either environmental/ chemical influence or certain variation that might lead to mutation in plant genome. To confirm mutation, in this experimentation, this morphological analysis was confirmed by molecular analysis. For that, the genomic DNA was extracted from both the plants and subjected to RAPD analysis. The morphological variants were shown distinct variation in DNA pattern by selected primers. Thus, RAPD profile proves that there was mutation in plant genome. This result supports the initiative to utilize morphological variants in plant breeding applications, and DNA fingerprinting.
Gasim, S; Elhassan, A M; Kharazmi, A;
production to Leishmania donovani and control antigens were measured in vitro using PBMC isolated at the time of diagnosis of kala-azar, after treatment of visceral leishmaniasis, during follow up, and at the time of diagnosis of PKDL. Proliferative responses and interferon-gamma (IFN-gamma) production were...... low at diagnosis and increased after treatment of kala-azar in both patients who developed (group 1) and those who did not develop PKDL later (group 2). In group 1, development of PKDL was always associated by an increased PBMC response to Leishmania antigen in proliferation and IFN-gamma production...... assays. There were no differences in Leishmania antigen-induced production of IL-4, IL-5 and IL-10 between or within the two groups. We have previously shown that Leishmania parasites spread to the skin during visceral leishmaniasis and proposed that PKDL was the result of an immunological attack...
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.
Nualsri, C.; Junmag, S.; Eksomtramage, T.
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 vari...
Jemni Chibani; Mokhtar Trifi; Messaoud Mars; Néjib Hasnaoui
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 char...
Characterization of different strains of Escherichia coli were analyzed for which samples were collected from different areas of Bangalore and cultured in MacConkey agar. Strains were finally isolated and their biochemical characterization was done for their proper identification, extraction of the nuclear DNA and its purity was tested. Quality testing with the help of spectrophotometer has been carried out and finally the application of a random amplified polymorphic DNA (RAPD) analysis for ...
KUSWANHADI; MUDJI LASMININGSIH; FETRINA OKTAVIA
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 ...
Rafael Gustavo Ferreira Morales; Juliano Tadeu Vilela de Resende; Marcos Ventura Faria; Marcela Carvalho Andrade; Luciane Vilela Resende; Carla Andrea Delatorre; Paulo Roberto da Silva
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 mark...
Sharaf-Eldin, M A; Al-Tamimi, A; Alam, P; Elkholy, S F; Jordan, J R
The artichoke (Cynara scolymus L.) is an important food and medicinal crop that is cultivated in Mediterranean countries. Morphological characteristics, such as head shape and diameter, leaf shape, and bract shape, are mainly affected by environmental conditions. A molecular marker approach was used to analyze the degree of polymorphism between artichoke hybrid lines. The degree of genetic difference among three artichoke hybrids was evaluated using random amplified polymorphic DNA-PCR (RAPD-PCR). In this study, the DNA fingerprints of three artichoke lines (A13-010, A11-018, and A12-179) were generated, and a total of 10 decamer primers were applied for RAPD-PCR analyses. Polymorphism (16.66 to 62.50%) was identified using eight arbitrary decamers and total genomic DNA extracted from the hybrids. Of the 59 loci detected, there were 25 polymorphic and 34 monomorphic loci. Jaccard's similarity index (JSI) ranged between 1.0 and 0.84. Based on the unweighted pair group method with arithmetic mean (UPGMA) similarity matrix and dendrogram, the results indicated that two hybrids (A13-010 and A11-018) were closely related to each other, and the A12-179 line showed more divergence. When identifying correct accessions, consideration of the genetic variation and genetic relationships among the genotypes are required. The RAPD-PCR fingerprinting of artichoke lines clearly showed that it is possible to analyze the RAPD patterns for correlation between genetic means and differences or resemblance between close accessions (A13-010 and A11- 018) at the genomic level. PMID:26782491
Full Text Available 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.
Lima L.H.C.; Návia D.; Inglis P.W.; Oliveira M.R.V. de
In 1991, the poinsettia strain, silverleaf whitefly or B biotype of Bemisia tabaci was detected in Brazil. This variant is a far more serious agricultural pest than the previously prevalent non-B (BR) biotype. The correct identification of B. tabaci is problematic since it is highly polymorphic with extreme plasticity in key morphological characters that vary according to the host. RAPD-PCR was used to survey the B biotype and other biotypes of B. tabaci in Brazil. Whiteflies were collected f...
Muthusamy RAJASEKAR; Muthusamy THANGARAJ; Thathiredypalli R. BARATHKUMAR; Jayachandran SUBBURAJ; Kaliyan MUTHAZHAGAN
Lates calcarifer (Bloch 1790) is one of the major economically important cultivable fish species in India. In this study, three populations of L. calcarifer was selected to assess the genetic diversity. Of which, two wild (Mudaslodai, Muthupettai) and one captive (Mutukadu) population. The genetic diversity of three populations of this species was studied using Random Amplified Polymorphic DNA (RAPD) markers. Ten random primers were used for the assessment of their genetic diversity and const...
Vale, Luis; Dieguez, Rebeca; Sánchez, Laura; Martínez, Paulino; Viñas, Ana
Understanding the genetic basis of sex determination mechanisms is essential for improving the productivity of farmed aquaculture fish species like turbot (Scophthalmus maximus). In culture conditions turbot males grow slower than females starting from eight months post-hatch, and this differential growth rate is maintained until sexual maturation is reached, being mature females almost twice as big as males of the same age. The goal of this study was to identify sex-specific DNA markers in turbot using comparative random amplified polymorphism DNA (RAPD) profiles in males and females to get new insights of the genetic architecture related to sex determination. In order to do this, we analyzed 540 commercial 10-mer RAPD primers in male and female pools of a gynogenetic family because of its higher inbreeding, which facilitates the detection of associations across the genome. Two sex-linked RAPD markers were identified in the female pool and one in the male pool. After the analysis of the three markers on individual samples of each pool and also in unrelated individuals, only one RAPD showed significant association with females. This marker was isolated, cloned and sequenced, containing two sequences, a microsatellite (SEX01) and a minisatellite (SEX02), which were mapped in the turbot reference map. From this map position, through a comparative mapping approach, we identified Foxl2, a relevant gene related to initial steps of sex differentiation, and Wnt4, a gene related with ovarian development, close to the microsatellite and minisatellite markers, respectively. The position of Foxl2 and Wnt4 was confirmed by linkage mapping in the reference turbot map. PMID:24415295
Dergam Jorge A.; Paiva Samuel R.; Schaeffer Carlos E.; Godinho Alexandre L.; Vieira Fabio
In the Rio Doce basin of southeastern Brazil, the freshwater fish Hoplias malabaricus (trahira) is a widespread predatory characin and one of the few resilient native fishes in a highly impacted lake system. In order to test for genetic differentiation in populations within this basin and for biogeographic relationships among populations of this species in other basins, a study was conducted using RAPD-PCR analysis of Rio Doce samples (N = 63) and phylogeographic analyses with mitochondrial D...
Genomic DNAs were isolated from both male and female plants of Acer negundo L., by modified CTAB method of Doyle and Doyle for plant genomic DNA isolation and consequently tested with RAPD technique.Some sex-related specific oxyribonucleic acid bands were amplified bY a group of decamer oligonucleotide random primers. This indicated that some genetic marks related to sexes of the species were found, which laid a foundation for sapling sex identification of Acer negundo L.
Badirzadeh, Alireza; Mehdi MOHEBALI; Ghasemian, Mehrdad; Amini, Hassan; Zarei, Zabiholah; AKHOUNDI, Behnaz; HAJJARAN, Homa; Emdadi, Dariush; Molaei, Soheila; Kusha, Ahmad; ALIZADEH, Saber
Visceral leishmaniasis (VL) is endemic in Northwest and southern Iran. Reports of cutaneous leishmaniasis (CL) in Northwest areas are rare, and its etiological agents are unknown. In the current study, we report six CL and two post kala-azar dermal leishmaniasis (PKDL) cases caused by Leishmania infantum from endemic areas of VL in the Northwest. Smears were made from skin lesions of 30 suspected patients in 2002–2011, and CL was determined by microscopy or culture. Leishmania spp. were ident...
En este trabajo se presenta un análisis de la importancia otorgada a los juegos de azar y su carácter intuitivo para la enseñanza de probabilidad en la propuesta institucional para educación primaria en México, así como los primeros resultados de una actividad realizada con niños de primaria.
Gouri S. Bhunia; Shreekant Kesari; Nandini Chatterjee; Rakesh Mandal; Vijay Kumar; Pradeep Das
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 t...
Soares-Ramos Juliana R.L.
Full Text Available Herbaspirillum spp. are endophytic diazotrophic bacteria associated with important agricultural crops. In this work, we analyzed six strains of H. seropedicae (Z78, M2, ZA69, ZA95, Z152, and Z67 and one strain of H. rubrisubalbicans (M4 by restriction fragment length polymorphism (RFLP using HindIII or DraI restriction endonucleases, random amplified polymorphic DNA (RAPD, and partial sequencing of 16S rDNA. The results of these analyses ascribed the strains studied to three distinct groups: group I, consisting of M2 and M4; group II, of ZA69; and group III, of ZA95, Z78, Z67, and Z152. RAPD fingerprinting showed a higher variability than the other methods, and each strain had a unique electrophoretic pattern with five of the six primers used. Interestingly, H. seropedicae M2 was found by all analyses to be genetically very close to H. rubrisubalbicans M4. Our results show that RAPD can distinguish between all Herbaspirillum strains tested.
S. M. Sarid Ullah
Full Text Available In the present study an efficient and easy method was followed for the isolation of DNA from meristem cylinder in five chewing sugarcane varieties, namely Amrita, Bomaby, Babulal (Co.527, Q83 and Misrimala. The quality and quantity of DNA were assured by visual estimation using agarose gel electrophoresis and UV spectrophotometry. The highest amount of DNA was retrieved from the Amrita (3250 ng/ml and the lowest amount was attained from the variety Q83 (1450 ng/ml. The amount of recovered DNA was enough for PCR amplification and marker studies such as random amplified polymorphic DNA (RAPD. Using RAPD markers, bands obtained from fingerprinting (190 bp to 1200 bp showed 73.5% polymorphism. The dendrogram, based on linkage distance using unweighted pair group method of arithmetic means (UPGMA, indicated segregation of the five chewing varieties of sugarcane into two main clusters. Amrita, Bombay and Misrimala were grouped in cluster 1 (C1 followed by sub-clusters. Babulal and Q83 were grouped in cluster 2 (C2. The results of the present investigation also revealed that the twenty RAPD primers were able to identify and classify the chewing sugarcane varieties based on their genetic relationship.
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
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.
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
Menochilus sexmaculatus commonly known as six spotted zig zag ladybird, is an aphidophagus and the most misidentified Coccinellids due to the occurrence of numerous color variants. The correct identification of Menochilus sexmaculatus and its strains is necessary to implement the use of biological control. In the present study phenotypic and genotypic polymorphism was investigated in Menochilus sexmaculatus collected from Punjab, NWFP and Sindh provinces of Pakistan. Six different morphs of the species were distinguished by analyzing its Elytral color and spot pattern and then Polymerase Chain Reaction was used to generate random amplification of polymorphic DNA (RAPD) from six different types of Menochilus sexmaculatus. Forty primers (OPA and OPC Kit) were used to perform RAPD PCR on six different types of Menochilus sexmaculatus of which, seven primers revealed different patterns related to the Menochilus sexmaculatus types. These seven primers (OPA-04, OPA-09, OPA-18, OPC-04, OPC-12, OPC-15 and OPC-18) produced 111 clear polymorphic bands and 6 scorable strain specific markers. The cluster analysis applied to RAPD data showed high polymorphism among six types and it can be concluded that these six types are six polymorphic strains of the same species, which has implications for natural and biological control of aphids. (author)
Molecular markers such as restriction fragment length polymorphisms (RFLPs) and polymerase chain reaction (PCR) based markers such as random amplified polymorphic DNAs (RAPDs) are being widely used for genetic analysis and DNA fingerprinting. In addition to biochemical markers for isozyme analysis, combined with morphological, physiological and pest and disease reactions, we have established RAPD markers as tools for assessing the genetic diversity in cassava (Manihot esculenta Crantz L.) and yam (Dioscorea species). In cassava, a preparatory study was conducted to characterize 24 cassave elite clones. A preliminary survey of 60 different commercially available primers from Operon, Alameda, California, United States of America (OPA, B and C series) was performed to identify those primers that gave good DNA fingerprinting. The results obtained show a relatively low degree of polymorphism. Nevertheless, the detected polymorphism allowed us to distinguish clones from one another. In yam (Dioscorea species), PCR-RAPD analysis was carried out to detect DNA polymorphism in the improved clones and land races of cultivated yams D. rotundata as well as in 11 wild species of the genus Dioscorea from Africa. Eight primers from the OPB and C series revealed polymorphism in the International Institute of Tropical Agriculture elite clones and land races. Two random primers (OPB-1 and OPB-2) gave excellent scorable polymorphism, allowing us to detect 17 different fragments that were scored as unit characters and used to draw phylogenetic relationships in Dioscorea using the computer program PAUP (phylogenetic analysis using parsimony)
Full Text Available 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 Jaccards 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.
The present study was conducted with an aim to analyze the genetic diversity and phylogenetic relationship among twenty five date palm (Phoenix dactylifera L.) cultivars from Pakistan. The leaf samples for DNA isolation were collected from four provinces of Pakistan i.e., Sindh, Punjab, Khyber Pakhtoonkhwa and Balochistan. DNA was isolated by modified CTAB method. The study was carried out by using six universal RAPD primers. The RAPD primers showed polymorphism among all date palm cultivars. The bands obtained were successfully used to differentiate the genotypes. Based on the pair-wise comparison of amplification products, the genetic relationship was estimated. All date palm cultivars showed variation at the DNA level. The average of genetic diversity among the date palm cultivars ranged 79.4%. A dendrogram was constructed using NTSYSpc programme, on the basis of this analysis, the populations were clustered into two main clusters and sub-clusters. Therefore, the high level of polymorphism was detected and its reproducibility suggests that RAPD markers are powerful and reliable tool for genetic diversity analysis of date palm cultivars. (author)
Full Text Available Genetic variability and differentiation among five annual caraway (Carum carvi populations originating from Tunisia, Germany and Egypt were examined for the first time. Random Amplified Polymorphic DNA (RAPD marker data were obtained and analysed with respect to genetic diversity, population structure and gene flow. Fourteen primers generated a total of 136 discernible and reproducible bands across the analyzed populations, out of which 56 were polymorphic. The UPGMA cluster analysis permitted the discrimination of all the genotypes and their sorting into 3 main groups. German and Egyptian caraway populations diverged significantly from Tunisian ones. Population clustering was made dependently from geographic origin. This has been further explained at the DNA level as we were able to select a set of RAPD fingerprints unique to each of the studied populations. Furthermore, dimensional graph derived from factorial analysis of RAPD frequency data, allowed significant grouping of the genotypes into five sub-plots, representing each one population. Shannon’s index values showed that variation ranks between rather than within populations. These results indicated that considerable genetic differences among C. carvi populations were registered.
Orozco-Castillo, C; Chalmers, K J; Waugh, R; Powell, W
RAPD (randomly amplified polymorphic DNA) markers generated by arbitary decamers have been successfully employed to detect genetic polymorphisms between coffee species and between Coffea arabica genotypes. The RAPD profiles were used to construct dendrograms and these were consistent with the known history and evolution of Coffea arabica. Material originating from Ethiopia and the arabica sub-groups - C. arabica var. typica and C. arabica var. bourbon - were clearly distinguished. RAPD analysis therefore reflects morphological differences between the sub-groups and the geographical origin of the coffee material. Species-specific amplification products were also identified, but, more importantly, amplification products specific to C. canephora were identified in two C. arabica genotypes, Rume Sudan and Catimor 5175. This diagnostic product is therefore indicative of interspecific gene flow in coffee and has biological implications for selective introgressive hybridisation in coffee. Our study demonstrates the power of the polymerase chain reaction technology for the generation of genetic markers for long-lived perennial tree and bush crops. PMID:24190527
João Filipi Rodrigues Guimarães
Full Text Available Genetic diversity in a collection of 64 sugar apple accessions collected from different municipalities in northern Minas Gerais was assessed by RAPD analysis. Using 20 selected RAPD primers 167 fragments were generated, of which 48 were polymorphic (28.7% producing an average of 2.4 polymorphic fragments per primer. Low percentage of polymorphism (< 29% was observed by using the set of primers indicating low level of genetic variation among the 64 accessions evaluated. Genetic relationships were estimated using Jaccard's coefficient of similarity. Accessions from different municipalities clustered together indicating no correlation between molecular grouping and geographical origin. The dendrogram revealed five clusters. The first cluster grouped C19 and G29 accessions collected from the municipalities of Verdelândia and Monte Azul, respectively. The second cluster grouped G16 and B11 accessions collected from the municipalities of Monte Azul and Coração de Jesus, respectively. The remaining accessions were grouped in three clusters, with 8, 15 and 37 accessions, respectively. In summary, RAPD showed a low percentage of polymorphism in the germplasm collection.
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
XIELei; LILiang—Qian; ZHANGDa—Ming
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.
Sugar cane is the fourth most important cash crop of Pakistan and is grown on 1 million hectares of land, with a total production of 37 million tonnes. It does not flower under existing environmental conditions. Sugar cane is vegetatively propagated and the national breeding programmes is restricted to the adaptation and multiplication of exotic varieties. Random amplified polymorphic DNA (RAPD) markers were used to establish polymorphisms among various local sugar cane varieties. DNA from the varieties L-118, L-116, BL-4, BF-162, Col-44, Col-54, Triton and Puri was isolated and amplified by polymerase chain reaction using ten nucleotide primers. The amplification profiles of all the sugar cane varieties were compared and the polymorphisms detected. DNA was isolated from the embryogenic calli of sugar cane subjected to gamma irradiation at different doses (0, 0.5, 2.0, 4.0 and 6.0 krad) and salt stresses (NaCl: 0, 50, 100, 150 and 200 mM), and was amplified with random primers to detect the polymorphisms introduced by stress. The banana is another important vegetatively propagated crop in Pakistan. DNA isolation from micropropagated banana was optimized and RAPD analysis performed on several clones of the banana variety Williams. The level of genetic variability revealed from calli and vegetatively propagated sugar cane and banana by RAPD analysis is discussed. (author). 10 refs, 5 figs, 2 tabs
Dergam Jorge A.
Full Text Available In the Rio Doce basin of southeastern Brazil, the freshwater fish Hoplias malabaricus (trahira is a widespread predatory characin and one of the few resilient native fishes in a highly impacted lake system. In order to test for genetic differentiation in populations within this basin and for biogeographic relationships among populations of this species in other basins, a study was conducted using RAPD-PCR analysis of Rio Doce samples (N = 63 and phylogeographic analyses with mitochondrial DNA (mtDNA haplotypes, including the Rio Grande and Macacu river basins. In the Rio Doce basin, the patterns of genetic similarity of RAPD-PCR markers (individual fingerprinting and Nei?s genetic distance suggest the existence of two genetically different groups, one composed of the lacustrine populations Carioca and Dom Helvécio, and the other of riverine and the remaining lacustrine populations. The differences in the RAPD-PCR patterns may be explained by the existence of sub-basins within this lacustrine system. A maximum parsimony tree of cytochrome b fragment (383 base pairs supports the view that trahiras of the Rio Doce share a complex biogeographic history with those of neighboring basins. The phylogeographic patterns may be explained by a common history of the watersheds of the Rio Doce, Paraíba do Sul, and Rio Grande basins, corroborating the hypothesis of a Plio-Pleistocene separation of these drainage systems, forming the Mantiqueira "divortium aquarium".
Prasad M P
Full Text Available BACKGROUD and OBJECTIVE: Piper, the pepper plants or pepper vines, are the genus of the Piperaceae family and are important economically and ecologically. It contains more than 1,000 species of shrubs, herbs and lianas, many of which are keystone species in their native habitat. The diversification of this taxon is of interest to understand the evolution of plants. In the scope of bioactive compounds, Piper species (P. nigrum, P. retrofractum and P. longum were screened for phytochemicals. METHODOLOGY: Phytochemical analysis of these plants showed presence of phenolic compounds, anthraquinones, terpenoids, flavinoids and lignin. Also, the methanolic extracts of these plants were tested for their antioxidant potential by free radical scavenging activity by DPPH assay. All the tested plants showed antioxidant potential and the overall antioxidant activity of P. nigrum was found to be highest. Genomic DNA was extracted from the fresh leaves of selected cultivars and PCR was performed by using RAPD primers to check the genetic diversity among these cultivars. From the PCR generated fingerprint, dendrogram was plotted by cluster analysis of similarity matrix. RESULTS: Dendrogram constructed by cluster analysis of RAPD markers showed that Piper nigrum and Piper retrofractum are closely related. Since morphological differences among these species are indistinctive, RAPD characterization can be helpful in their Discrimination. CONCLUSION: This finding can be used as prerequisite for plant breeding activities as well as for conservation of genetic resources.
John Yew Huat Tang; Mohd Ikhsan Khalid; Syazana Aimi; Che Abdullah Abu-Bakar; Son Radu
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.
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.
A. M. Musa
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.
ShivaRajAdhikari; SiripenSupakankunti; MMahmudKhan
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.
Eltahir Awad Gasim Khalil
Full Text Available Visceral leishmaniasis (VL is a parasitic disease characterized by immune suppression. Successful treatment is usually followed by immune reconstitution and a dermatosis called post-Kala-azar dermal leishmaniasis (PKDL. Recently, PKDL was described as one of the immune reconstitution syndromes (IRISs in HIV/VL patients on HAART. This study aimed to present PKDL as a typical example of paradoxical IRIS in non-HIV/AIDS individuals. Published and new data on the pathogenesis and healing of PKDL was reviewed and presented. The data suggested that PKDL is a typical example of paradoxical IRIS, being a new disease entity that follows VL successful treatment and immune recovery. PKDL lesions are immune inflammatory in nature with granuloma, adequate response to immunochemotherapy, and an ensuing hypersensitivity reaction, the leishmanin skin test (LST. The data also suggested that the cytokine patterns of PKDL pathogenesis and healing are probably as follows: an active disease state dominated by IL-10 followed by spontaneous/treatment-induced IL-12 priming, IL-2 stimulation, and INF-γ production. INF-γ-activated macrophages eliminate the Leishmania parasites/antigen to be followed by LST conversion and healing. In conclusion, PKDL is a typical example of paradoxical IRIS in non-HIV/AIDS individuals with anti-inflammatory cytokine patterns that are superseded by treatment-induced proinflammatory cytokines and lesions healing.
Jónathan Hernández Núñez; Magel Valdés Yong; Andrés Oscar Torres Chávez
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...
A rapid immunochromatographic dipstick test (ICT) has become available for the qualitative detection of anti-Leishmania antibody using recombinant rK39 antigen. This study was carried out at the department of Microbiology of Rajshahi Medical College, Bangladesh, in order to evaluate the diagnostic potential of immunochromatographic dipstick test. A total of one hundred cases including 60 admitted patients with strong clinical suspicion of kala-azar and 40 healthy controls were investigated for the performance of the ICT. Splenic smears were examined for microscopic detection of Leishman Donovan (LD) bodies obtained from the admitted patients only and smear-positive cases were considered as gold standard as well as confirmed cases of kala-azar. Out of 60 suspected patients, fifty three (88.33%) were found smear-positive and fifty nine (98.33%) were positive for immunochromatographic strip test. All smear-positive cases were also positive for strip test. Voluntary healthy controls (40), which included twenty persons from the endemic zone and twenty from non-endemic zone of kala-azar, were found all negative for the strip test. The sensitivity and specificity of immunochromatographic strip test were found to be 100.00% and 86.95% respectively. Present study findings again reinforce that the immunochromatographic strip test is a simple, reliable and easy-to-perform non-invasive diagnostic tool for visceral leishmaniasis in the endemic area of Bangladesh. (author)
Claris B. Palatnik de Sousa
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.
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.
Boris Gutarra Castillo
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.
Manuel TORAL IBA(N)EZ; Margarita CARU; Miguel A.HERRERA; Luis GONZALEZ; Luis M.MARTIN; Jorge MIRANDA; Rafael M.NAVARRO-CERRILLO
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.
Jiang Changshun; Zou Dongmei; Zhang Yizheng
Stylosanthes guianenesis Sw. is an important tropical forage legume grown and utilized in the tropics and the subtropics of South China. Anthracnose, caused by Colletotrichum gloeosporioides (Penz.) Sacc., is a major constraint to the extensive use of Stylosanthes. Forty-five accessions of S. guianensis were assessed with RAPD for genetic diversity and for resistance to anthracnose. RAPD analysis was performed using twenty primers screened from 200 arbitrary oligonucleotides, and a 71.5% level of polymorphism was found. The dendrogram obtained with unweighted pair group method of averages (UPGMA) based on the RAPD data showed genetic similarity from 50% to 94% among all stylo accessions, and fourteen clusters were defined at 66.5% genetic similarity. Two strains of C. gloeosporioides from stylo in China were used for anthracnose resistance screening. All plant accessions showed variation in the reaction to two strains and the correlation of resistance had a value of 0.904. Multiple correspondence analysis displayed a random distribution of the resistance or susceptibility response with respect to the genetic diversity measured by RAPD analysis except one group. Mean distance was also calculated to determine the diversity within clusters. From our results, the RAPD analysis is an effective and efficient technique of providing quantitative estimates of genetic similarity among stylo accessions.
The project was carried out in 2007 with the purpose of consideration for using the two simple and inexpensive molecular techniques to estimate changes in DNA of rice mutant after gamma irradiation. Three rice cultivars: Basmati370, Tam Thom (TT1), IR64 and three gamma irradiated mutants BDS, TDS and VND 95-20 respectively, were used. Suitable DNA extraction procedure was obtained. PCR optimization was conducted on three important factors including: amount of MgCl2, DNA concentration and annealing temperature. 2.5 mM of MgCl2 for RAPD-PCR and 3.75 mM for RAMP-PCR were found the best. 40 ng DNA provided a good amplification for RAMP-PCR; this figure was 50 ng for RAPD-PCR. Annealing temperatures were determined at 36 oC for RAPD primer and at 55±3 oC for Microsatellite primer. Final results showed that, both RAPD-PCR and RAMP-PCR could detect changes in DNA of rice mutants after gamma irradiation compared to their parents. Percentage of DNA changes determined by RAPD-PCR and RAMP-PCR on Basmati370 and its mutant BDS were 11.49% and 21.2% respectively; These on TT1 and TDS were 8.98% and 15.4%; and on IR64 and VND 95-20 were 3.45% and 4.95%. (author)
Kaushal, H; Bras-Gonçalves, R; Avishek, K; Kumar Deep, D; Petitdidier, E; Lemesre, J-L; Papierok, G; Kumar, S; Ramesh, V; Salotra, P
Post-kala-azar dermal leishmaniasis (PKDL) is a chronic dermal complication that occurs usually after recovery from visceral leishmaniasis (VL). The disease manifests into macular, papular and/or nodular clinical types with mono- or polymorphic presentations. Here, we investigated differences in immunological response between these two distinct clinical forms in Indian PKDL patients. Peripheral blood mononuclear cells of PKDL and naive individuals were exposed in vitro to total soluble Leishmania antigen (TSLA). The proliferation index was evaluated using an enzyme-linked immunosorbent assay (ELISA)-based lymphoproliferative assay. Cytokines and granzyme B levels were determined by cytometric bead array. Parasite load in tissue biopsy samples of PKDL was quantified by quantitative polymerase chain reaction (qPCR). The proportion of different lymphoid subsets in peripheral blood and the activated T cell population were estimated using flow cytometry. The study demonstrated heightened cellular immune responses in the polymorphic PKDL group compared to the naive group. The polymorphic group showed significantly higher lymphoproliferation, increased cytokines and granzyme B levels upon TSLA stimulation, and a raised proportion of circulating natural killer (NK) T cells against naive controls. Furthermore, the polymorphic group showed a significantly elevated proportion of activated CD4(+) and CD8(+) T cells upon in-vitro TSLA stimulation. Thus, the polymorphic variants showed pronounced cellular immunity while the monomorphic form demonstrated a comparatively lower cellular response. Additionally, the elevated level of both activated CD4(+) and CD8(+) T cells, coupled with high granzyme B secretion upon in-vitro TSLA stimulation, indicated the role of cytotoxic cells in resistance to L. donovani infection in polymorphic PKDL. PMID:26948150
RONAN XAVIER CORRÊA
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
Patrícia da Silva Nascente
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
Tulsiani, Suhella; Craig, S B; Graham, G C;
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 as ...
Diversidade genética de Chenopodium ambrosioides da região cacaueira da Bahia com base em marcadores RAPD Genetic diversity based on RAPD markers of Chenopodium ambrosioides from the cocoa region of Bahia State, Brazil
Simone Gualberto Santos; Ronan Xavier Corrêa
Chenopodium ambrosioides L., conhecida no Brasil por suas propriedades medicinais e usada principalmente para o controle de verminoses intestinais, é pouco estudada quanto à diversidade genética. O objetivo deste trabalho foi avaliar a diversidade genética de 16 indivíduos de C. ambrosioides, provenientes de diferentes municípios da região cacaueira da Bahia, pela técnica de RAPD (DNA polimórfico amplificado ao acaso). Apenas 6,9% das 216 bandas RAPD amplificadas foram polimórficas e a anális...
DING Wei; ZHANG Zhengwang; CHANG Jiang; ZHANG Er; WU Xiushan; ZHANG Jinguo
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.
Manica-Cattani, M F; Zacaria, J; Pauletti, G; Atti-Serafini, L; Echeverrigaray, S
Twenty-seven accessions of Lippia alba Mill. collected in Rio Grande do Sul state, Brazil, were analysed by ISSR and RAPD markers to evaluate their genetic variability and relationships. Six ISSR primers and four RAPD primers generated 120 amplified fragments, most of which were polymorphics. The overall genetic variability among accessions was very high when compared with other plant species. The hierarchical analysis of molecular data (UPGMA) showed low relationship between accessions, and no grouping between accessions of the same chemotype. Canonical functions allowed identifying some variables related with the chemical characteristics of the essential oils. Both ISSR and RAPD markers were efficient to address the genetic diversity of L. alba, and may contribute to the conservation and breeding of this increasingly important aromatic and medicinal species. PMID:19675941
Meera N., Lohithaswa HC, Niranjana Murthy and Shailaja Hittalmani
Full Text Available Genotype specific fingerprints were detected in two grain amaranthus varieties KBGA1 and Suvarna using SSR and RAPD markers. In this study 41 Pigeon Pea SSR markers and 6 RAPD markers were used to generate DNA fingerprints for the two varieties of grain amaranthus. Analysis of polymorphic fragments generated from SSR and RAPD markers revealed the genetic variation between grain amaranthus varieties KBGA1 and Suvarna. The results indicate that DNA markers are appropriate tools for assessing genetic variation within and between the species of amaranthus and suggest that cultivated varieties of Amaranthus have significant genetic variation. The differences generated by the markers can be used as fingerprints for detecting the varieties. This is the first report of the utilization of legume microsatellite markers in Amaranthus.
Use of random amplified polymorphic DNA (RAPD to detect DNA damage induced by Prangos ferulacea (Umbelliferae essential oil against the Mediterranean flour moth Ephestia kuehniella Zeller (Lepidoptera: Pyralidae
Ercan Fahriye Sumer
Full Text Available DNA damage detection in Ephestia kuehniella by RAPD-PCR Abstract - The random amplified polymorphic DNA (RAPD assay is a useful method for detecting genotoxin-induced DNA damage. In the present study, this assay was evaluated to measure the essential oil of Prangos ferulacea-induced DNA changes in Ephestia kuehniella Zeller larvae. The third instar larvae of E. kuehniella were exposed to 500 μl L-1 essential oil for 24 h. Forty random primers were used for RAPD-PCR of which eleven produced unique polymorphic band patterns. In comparison with control larvae, the essential oil-treated larvae showed greater changes in RAPD profiles. This is the first report of an analysis of the genotoxic effect of P. ferulacea essential oil against E. kuehniella larvae using RAPD-PCR.