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.
Full Text Available Turkish Red Pine (Pinus brutia Ten. is an important forest tree species in Turkey for various economic and ecological reasons. In this study, nine RAPD (Randomly Amplified Polymorphic DNA primers were used to determine genetic variation within and among populations of P. brutia located at the Duzlercami common-garden test site. This site was established in 1979 and includes six natural populations of P. brutia from two altitudinal transects extending from the coast to higher elevations in the Antalya region of Turkey. A total of 32 polymorphic RAPD loci were found in the analyzed six populations. The mean proportion of polymorphic loci among population samples equals 100%, mean number of alleles for each locus = 2.0, effective allele number = 1.71, Shannon�s information index = 0.58, and mean Nei (1973�s gene diversity value = 0.4. According to GST results, a high proportion of genetic diversity (95-99% is found within populations. A relatively high genetic differentiation was found among altitudinal population pairs in both transect. Also, data on quantitative traits (total height and/or diameter at different ages (13, 17, 30 years were compared with molecular data. There are similarities between the results obtained from RAPD markers and those obtained from the quantitative traits. The differentiation in quantitative traits appears to be due to local adaptation of populations. Data suggest that priority should be given to the selection of material based on geographic origin along the altitudinal gradients of P. brutia populations to conserve the genetic resource of species.
CAO Shao-xian; YANG Li-guo; JIANG Xun-ping; LIU Hong-lin; LU Wei-zhong; XIANG Yang-hai
Blood and tissue samples were collected from 105 goats including 60 Boer goats (30 for eachsex), 30 Xuhuai goats (15 for each sex) and 15 Haimen goats (7 stud and 8 does). DNA was extracted andDNA pools were constructed on the basis of goat breeds. In 36 selective primer combinations, 29 combinationsamplified totally 3 253 markers including 92 polymorphic markers by amplified fragment length polymor-phism (AFLP). On average, 3.17 polymorphic markers were amplified per combination, with a polymorphicfrequency of 2.8%. The primer combinations amplifying more polymorphic markers (showed in brackets)were involved in E00+ACG/M00+CAA (13), E00+ACG/M00+CAG (10), E00+AAC/M00+CAC (8)and E00+AAC/M00+ACT (7). A total of 183 markers including 60 polymorphic markers were amplified byRAPD from the pooled DNA of three breeds using 22 primers with strong polymorphism and high reproduc-ibility selected from 93 RAPD primers. On average, 2.73 polymorphic markers were amplified per primer,with a polymorphic frequency of 32.8%. The results of AFLP and RAPD coincidently suggested that the ge-netic distance is the closest between Xuhuai and Haimen goat, next between Xuhuai and Boer goat, and the far-thest between Haimen and Boer goat. According to the UPGMA method, Haimen and Xuhuai goats can begathered together as a cluster, then Boer goat. Both methods can be used to implicate the genetic difference ofthese three breeds, in particular AFLP has more polymorphic markers.
Full Text Available Acid lime (Citrus aurantifolia Swingle is an important commercial fruit crop, cultivated from terai to high hill landscapes of Nepal. However, production and productivity is very low due to various reasons including infestations by various diseases and pests, lack of diseases and pests resistant and high yielding varieties. In this context, determination of genetic variation at molecular level is fundamental to citrus breeders for the development of elite cultivars with desirable traits. In the present study, Random Amplified Polymorphic DNA (RAPD marker technique has been employed to assess genetic diversity in 60 acid lime landraces representing different agro-ecological zones of eastern Nepal. Nine selected arbitrary primers generated 79 RAPD fragments of which 75 were polymorphic (94.94%. Phenogram was constructed by NTSYSPC ver. 2.21i using UPGMA cluster analysis based on Jaccard’s similarity coefficient to deduce overall genetic diversity and relationships of the acidlime genotypes under study. Sixty acid lime landraces formed seven clusters and similarity value ranged from 38% to 98% with an average of 72%. Genetic variation at different agro-ecological zones was assessed using Popgene ver. 1.32 and found 47% to 69.6% polymorphism. Shannon’s index and Nei’s gene diversity showed highest level of acid lime diversity in Terai zone (PPB, 69.62%; H, 0.213; I, 0.325 followed by mid-hill zone (PPB, 67.09%; H, 0.208; I, 0.317. The results obtained will be useful to citrus breeders for elite cultivar development. The RAPD-PCR technique is found to be the rapid and effective tool for genetic diversity assessment in acid lime landraces of Nepal.
Full Text Available Oktavia F, Lasminingsih M, Kuswanhadi. 2011. Selection of parent trees for Rubber (Hevea brasiliensis breeding based on RAPD analysis. Nusantara Bioscience 3: 124-129. The parent trees’ clones usually originate from the previous generation having the potential of high production with better agronomical characters. Although, phenotype characters can determine the genetic variability among accessions, they are highly sensitive to environmental factors, so it is often difficult to identify the difference between closely related clones. The genetic variability or phylogenetic relationships among rubber clones can be analysis using RAPD method, and based on the result, the parent trees can be selected. This research was aimed to analyze the genetic distance among rubber clones using RAPD method. Analysis was conducted on 45 rubber clones with 12 random primers. Pair-wise comparisons of unique and shared polymorphic amplification products were used to generate similarity coefficients. These coefficients were employed to construct a dendogram by using an Unweighted Pair-Group Method with Arithmetical Averages (UPGMA. The amplification of genomic DNA from 45 clones yielded 2408 DNA fragments, ranging in size from 250 bp to 3000 bp. The range of genetic similarity matrix was very wide (59.18%-94.23%. It indicated that most of the clones have a low level of polymorphism. The lowest genetic similarity (59,18% was found between RRIC 110 and AVROS 352 clones, while the highest (94.23% was between IRR 41 and IRR 42 clones. Cluster analysis showed that 45 clones of rubber were divided into two groups, the biggest group consisted of 30 clones, while the other one consisted of 15 clones with a genetic similarity value of 0,73.
using primer peh no.6 (1.0kb) could be considered as diagnostic band as it was fixed in Leersia infesting populations and was always absent in rice infesting populations of N. lugens. Boostrapping analysis showed 100% confidence level for the separate clusterings between the rice and Leersia infesting populations of N. lugens and also for the genetically isolated group, N. bakeri. These results represent the evidence for genetic structuring within the species, N. lugens with high esterase activities usually caught off rice and those with low esterase activities usually caught off Leersia. As the morphological characters of stridulatory organs and acoustic signals differed in the two sympatric populations of N. lugens so there is an evidence of sibling species in brown planthopper, N. lugens complex. Our results in DNA fingerprinting were consistent with the results of Claridge et al., based on their nymphal survival, virulence, ovipositional preference, mate choice and hybridisation experiments. Using the diagnostic marker and the genetic distance between rice and Leersia infesting populations,as well as cluster analysis from LP-RAPD-PCR, suggested that brown planthopper with high esterase activity captured from rice and brown planthopper with low esterase activity captured from L. hexandra ar closely related sibling species. This information has practical implications for formulating effective control measures against N. lugens, which is a major pest of rice not only in Malaysia but also throughout southeastern Asia, southern Asia and Australia
Full Text Available Genetic variability and identification of some molecular markers were studied in twenty promising lines of wheat using agronomic traits, ISSR (inter simple sequences repeats and RAPD (random amplified polymorphic DNA markers. Significant variation was evidenced in all agronomic traits. The lines proved to be superior to the check cultivar Sahel1 in yield and its component traits. Lines L2, L7 and L8 were the best in most yield component traits in both seasons. Moreover, Lines L2, L4, L5, L7 and L8 showed drought tolerance by which they displayed high performance in agronomic traits as well as a low drought susceptibility index. The percentage of polymorphism was 39.3% and 53.2% for ISSRs and RAPDs, respectively. UBC-881 belonged to penta-nucleotide repeat sequences (GGGTG that produced the highest level of polymorphism, while UBC-846 belonged to di-nucleotide repeat sequences (CA that produced the lowest level of polymorphism. Genetic similarities among wheat lines based on ISSR and RAPD markers ranged from 0.81 to 1.00 and from 0.86 to 0.98, respectively. There was a low average of PIC (polymorphism information content values which were 0.10 (ISSR and 0.15 (RAPD. The RAPD technique exhibited a higher marker index (MI = 0.69 compared to ISSR (MI = 0.43. There was insignificant correlation between ISSR and RAPD data (0.168, p > 0.05. There were two markers (UBC-881450bp and OPF-10540bp, on each of which two traits regressed significantly. The associated markers each explained a maximum regression of 18.92–34.95% of the total available variation for individual associated traits.
The wheat mutant 92k809 with resistance to Fusarium gramine arum was selected by means of induction mutation of irradiation, tissue culture and cell screening. The mutant 92k809 has stable resistance to the disease, good agronomic traits with increasing yield of 4.9% when compared with its parent. The specific DNA fragments, which both 92k809 and resistant material Sumai No.3 own, have been fond by RAPD technique. These fragments, which do not exist in the parent, are related to scab of wheat
Liu Zhenhui; Kong Jie; Liu Ping; Deng Jingyao
The genetic structure of three stocks (K, H and S) of Penaeus chinensis was detected by RAPD method which useed 41 random oligonucleotide primers with 10 bases. Stock K was from the spawning stock of the western coastal waters of the Korea Peninsula, H was from the Huanghai SeaBohai Sea stock and S was from the health cultured stock in Jimo, Shandong Province, China. Three genetic markers, F- 02540, OPN - 13800 and OPN - 202200, which could differentiate the three different stocks, have been reported. The basic data for marker assisted selection (MAS) on genetic breeding of Penaeus chinensis are provided.
Carlos Bernard Moreno Cerqueira-Silva
Full Text Available This study aimed to evaluate the genetic diversity by RAPD markers in 20 genotypes of ‘yellow’ passion fruit (Passiflora edulis Sims. The 16 primers generated 92 markers, 57 (62% of which were polymorphic. The genetic distance (gdij estimated by the complement of the Dice index (gdij = 0.19 and genotype grouping based on UPGMA algorithm showed low variability among genotypes. These results show a narrower genetic basis than reported for other Passiflora populations and the need to increase this variability by germplasm introduction. Divergent genotypes were also identified for the choice of parents for crosses for genetic gains in traits previously selected within the population studied.
H. E. SAWAZAKI; M. L. A. BOVI; Sodek, L.; COLOMBO C.A.
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),...
Random amplified polymorphic DNA (RAPD) analysis was used to evaluate genetic diversity among eight garlic mutants resistant to white rot disease (Sclerotium cepivorum) and two controls. Twelve of 13 synthetic random primers were found to identify polymorphism in amplification products. Mutants characterised with moderate resistance to white rot were closely related to the control using cluster and correlation analyses. On the other hand, highly resistant mutants were quite distant from the control with low correlation coefficients. The banding patterns produced by primer OPB-15 (GGAAGGGTGTT) with highly resistant mutants may be used as genetic markers for early selection of resistant plants. (author)
Palm oil as a plant producing oil is one of the primadonna oil palm plantations that become a source of producer non-migas for Indonesia foreign exchange. Superior varieties of palm oil produced by various research institutions that is a hybrid is tenera dura x pisifera (DxP). Pisifera having an important function in the production of seed oil palm. Hence the population pisifera very important to be managed and developed . A method of RAPD is a method to identify a number of large polymorphis...
卢钢; 曹家树; 陈杭
A genetic linkage map comprised of 131 loci was constructed with an F2 population derived from an inter-subspecific cross between Brassica 'qisihai'. The genetic map included 93 RAPD loci, 36 AFLP loci and 2 morphological loci organized into 10 main linkage groups (LGs) and 2 small groups, covering 1810.9cM with average distance between adjacent markers being approximately 13.8cM. The map is suitable for identification of molecular markers linked to important agronomic traits, QTL analysis, and even for marker-assisted selection in breeding programs of Chinese cabbage and turnip.
Full Text Available Annually many numbers of pationts with Cutaneous Leishmaniasis (CL have been reported in Kermanshah province- IRAN. The study aimed to identify species of Leishmania isolated from patients with CT in Kermanshah. Seven isolates of Leishmania obtained from patients with CL, without any travelling to other provinces, were cultured in NNN medium. After mass production of leptomonads in RPMI 1640 medium DNA was purified and the species were diagnosed using RAPD-PCR technique. The study of electrophoretic fingerprints of the product of RAPD-PCR in seven isolates showed that Leshmania major was the causative agent of CL patients in Kermanshah province. More studies in this field recommended.
J. L. Valle
Full Text Available Se asoció Cenchrus ciliaris (Cc y Gliricidia sepium (Gs para evaluar la producción de biomasa y la composición química de los forrajes en la fase de establecimiento, en el estado de Morelos, México, en condiciones de trópico seco. Se empleó un análisis de varianza con diseño en bloques al azar, en donde T1 fue pasto solo, T2 Cc más Gs con 5,000 plantas ha-1 y el T3 Cs más Gs con 14,285 plantas ha-1. Se midió la producción de forraje individual y asociado con materia seca (t MS/Ha, la altura (A en cm, la proteína cruda (PC%, las fracciones de fibra (FDN% y FDA% y la digestibilidad in vitro de la materia seca (DIVMS%. El T3 tuvo una mejor producción de biomasa (P0.05. La asociación de 14,285 plantas ha-1 de G. sepium con pasto Cenchrus ciliaris mejoró la producción de biomasa y la disponibilidad total de nutrientes por superficie cultivada al establecimiento
Mohammad Shafiul Alam; Hirotomo Kato; Mizuho Fukushige; Yukiko Wagatsuma; Makoto Itoh
Mymensingh is the most endemic district for kala-azar in Bangladesh. Phlebotomus argentipes remains the only known vector although a number of sand fly species are prevalent in this area. Genotyping of sand flies distributed in a VL endemic area was developed by a PCR and restriction-fragment-length polymorphism (RFLP) of 18S rRNA gene of sand fly species. Using the RFLP-PCR analysis with AfaI and HinfI restriction enzymes, P. argentipes, P. papatasi, and Sergentomyia species could be identif...
Melaku, Yosef; Collin, S.; Keus, Kees; Gatluak, Francis; Ritmeijer, Koert; Davidson, R. N.
Médecins sans Frontières-Holland has treated > 67,000 patients with kala-azar (KA) in southern Sudan since 1989. In 2002, we replaced the standard regimen of 30 days of daily sodium stibogluconate (SSG) with a 17-day regimen of daily SSG combined with paromomycin (PM). We analyzed data for 4,263 primary KA patients treated between 2002 and 2005 in southern Sudan to determine the relative efficacy of the combination therapy regimen (PM/SSG). The initial cure rate among patients treated with PM...
Ismail, A; Khalil, E A G; Musa, A M;
Post kala-azar dermal leishmaniasis (PKDL) is a dermatosis caused by persistence of Leishmania donovani parasites in the skin following apparently successful treatment of visceral leishmaniasis. The distribution of PKDL lesions in Sudanese patients often mirrors the clothing habits of those...... affected. It is most severe in or confined to the sun-exposed parts of the skin. It is well established that elimination of Leishmania parasites requires activation of parasitised macrophages by a Th1 immune response and that the latter is depressed by ultraviolet light (UVB). In this paper, we...
Ismail, Ahmed; Gadir, A Fattah A; Theander, Thor G;
cells. Degenerating basal keratinocytes expressed HLA-DR, ICAM-1 and Leishmania antigen and closely interacted with CD4 T cells. Regional lymph nodes showed hyperplasia of the B- and T-cell zones. Conclusions: The inflammatory reaction in PKDL lesions is in response to Leishmania parasites and....../or antigen. The majority of cells are CD4 T cells. Degeneration of the basal keratinocytes is probably due to the action of cytotoxic CD4 T cells interacting with leishmania-expressing epidermal cells. Ismail A, Gadir AFA, Theander TG, Kharazmi A, El Hassan AM. Pathology of post-kala-azar dermal...
Immanuel Selvaraj. C, Pothiraj Nagarajan, K. Thiyagarajan, M Bharathi, and R Rabindran
Full Text Available Genetic diversity among 26 rice genotypes was investigated using RAPD markers. The genotypes were screened for the leaf blastdisease reaction at two different environments. The average number of alleles amplified per primer was 9.03. Average number ofpolymorphic bands per primer was 6.80 with average polymorphism information content (PIC of 0.264. Clustering based ondendrogram revealed two major clusters and 5 sub clusters. Principal Coordinate Analysis (PCoA revealed three major groups.The first coordinate does not discriminate any of the genotypes based on the geographical origin, but the second and thirdcoordinates differentiated South East Asian and South Asian genotypes clearly. Genetic diversity analysis of rice genotypes withRAPD marker system and phenotypic screening for blast resistance revealed that White Ponni (susceptible and Moroberekan(resistant were one among the genetically distant and contrasting parents for leaf blast resistance. There is no cleardiscrimination of the markers to distinguish leaf blast resistant and susceptible genotypes into separate clusters by the principalcoordinate analysis.
A reliable protocol for adult Quercus suber L. somatic embryogenesis (SE) was developed recently. To evaluate the potential use of this protocol in cork oak forest breeding programs, it is essential to guarantee somatic embryos/emblings genetic stability. Random Amplification of Polymorphic DNA (RAPD) is currently used to assess somaclonal variation providing information on genetic variability of the micropropagation process. In this work, SE was induced from adult trees by growing leaf explants on MS medium supplemented with 2,4-D and zeatin. Embling conversion took place on MS medium without growth regulators. DNA from donor tree, somatic embryos and emblings was used to assess genetic variability by RAPD fingerprinting. Fourteen primers produced 165 genetic loci with high quality and reproducibility. Despite somatic embryos originated some poor quality PCR-profiles, replicable and excellent fingerprints were obtained for both donor plant and embling. Results presented no differences among regenerated emblings and donor plant. Hence, the SE protocol used did not induce, up to moment, any genetic variability, confirming data previously obtained with other molecular/genetic techniques, supporting that this protocol may be used to provide true-to-type plants from important forestry species. (author)
El-Banna, A N; El-Nady, M F; Dewir, Y H; El-Mahrouk, M E
Fasciated and normal stem segments of Opuntia microdasys, Opuntia cylindrica, Huernia primulina and Euphorbia lactea were collected from the same plant and compared for their anatomy, water relations and genetic variations. Anatomical differences in terms of thickness of cuticle, vascular bundle, xylem and phloem were analyzed in both normal and fasciated stems. The mucilage cells were higher in the fasciated form of Opuntia microdasys than that in the normal form. Water status in terms of total water content (TWC), water deficit and relative water content (RWC) was influenced by fasciation. Genetic variations were tested in normal and fasciated stems using randomly amplified polymorphic DNA (RAPD) fingerprints and SDS-PAGE of soluble protein extracts. SDS-PAGE protein and RAPD analysis confirmed that normal and fasciated tissues were genetically different. Polymerase chain reaction (PCR) yielded different polymorphic banding patterns that were unique to each primer and distinguishable over all samples. The PCR results of normal and fasciated samples were significantly different in cases of primers P1, P2 and P3. These results indicate that occurrence of fasciation in Opuntia microdasys, Opuntia cylindrica, Huernia primulina and Euphorbia lactea is an epigenetic mutation of tissues. PMID:24013892
Rout, Gyana Ranjan
Identified germplasm is an important component for efficient and effective management of plant genetic resources. Traditionally, plant identification has relied on morphological characters like growth habit, floral morphology like flower colour and other characteristics of the plant. Studies were undertaken for identification and genetic variation within 15 clones of Tinospora cordifolia through random amplified polymorphic DNA (RAPD) markers. Analysis was made using forty decamer primers. Out of them, 15 primers were selected and used for identification and genetic relationships within 15 clones. A total of 138 distinct DNA fragments ranging from 0.2 to 3.2 kb were amplified using 15 selected random primers. The genetic similarity was evaluated on the basis of presence or absence of bands. The genetic distance was very close within the clones. Thus, these RAPD markers have the potential for identification of species and characterization of genetic variation within the population. This study will be helpful to know the genetic background of the medicinal plants with high commercial value, and also provides a major input into conservation biology. PMID:16610228
Full Text Available RAPD analysis of nine Thai pineapple cultivars, including 'Phulae', 'Sawee', 'Tradsithong', 'Phuket', 'Pattavia', 'Intrachitdang', 'Intrachitkhow', 'Petburi No.1', and 'Nanglae', showed that, of 40 arbitrary 10- mer primers, 17 primers gave 206 DNA fragments ranging from 510 to 4,700 bp. One hundred and forty-five (70.4% of the amplified fragments were polymorphic. RAPD analysis using NTSYS-pc Version 2.01e also showed that the similarity coefficients among the cultivars were 0.643-0. 963. The dendrogram indicated that the cultivars were clustered into 3 groups, consistent with the morphological data. The first group, consisting of 'Phuket', 'Phulae', 'Tradsithong', 'Sawee', and 'Petburi No.1', had morphological characteristics of the Queen group, while those of the second ('Intrachitdang' and 'Intrachitkow' and the third ('Nanglae' and 'Pattavia' groups could be determined morphologically to be members of the Spanish and Cayenne groups, respectively. 'Intrachitdang' and 'Intrachitkow' have similarity coefficient of 0.963, while that of 'Phulae' and 'Phuket' is 0.950. These pairs of cultivars are probably the same cultivars. The morphological differences between them are probably caused by mutations, differences in environment and agricultural practices, or combinations of these factors.
The taxonomic relationships among 25 samples representing nine species of Orobanche L. (Orobanchaceae) were determined by the analysis of morphological characters and molecular polymorphism using random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR). In order to construct dendrogram elucidating the relationships among the examined taxa, the coded data were analyzed using the software package NTSYS-pc 2.1 based on the Neighbor-joining (NJ) tree building method based on a distance matrix. The aim of this study is to develop taxonomic relationship based on morphological and molecular data, in order to obtain a more reliable taxonomic relationship of Orobanche species under study. The dendrogram produced by the analysis of the molecular data (RAPD and ISSR) resembled that constructed by NJ dendrogram for the morphological variation. The studied taxa were separated in two groups, the first comprised of the five species of section Trionychon (O. purpurea, O.lavandulacea, O. ramosa, O. mutelii and O. aegyptiaca) and the second comprised of the four species of section Orobanche (O.cernua, O. crenata, O. minor and O. pubescens). High similarity was detected between O. pubescens and O. minor. The results confirmed the close relationship between O. ramosa and O. mutelii. Moreover, this study demonstrated the grouping of the studied taxa in most cases by geographically isolated population. (author)
You, Feng; Wang, Wei; Xu, Dongdong; Zhu, Xiangping; Ni, Jing; Wu, Zhihao; Xu, Yongli; Wang, Xincheng; Zhang, Peijun
The hybrid between olive flounder Paralichthys olivaceus and stone flounder Kareius bicoloratus was produced by artificial insemination of olive flounder eggs with stone flounder sperm. Sinistral and dextral are two types of hybrid progeny after metamorphosis. Karyotypes of both hybrid flounders are the same as those of the two parental species. Of the 22 loci examined from 12 allozymes, 12 confirmed hybridization of the paternal and maternal loci in hybrids and no difference was found in allozyme patterns of sinistral and dextral hybrid fishes. RAPD patterns of these specimens were also studied with 38 primers selected from 104 tested. Among them, the PCR products of 30 primers showed hybridization of the paternal and maternal bands. Genetic variation between hybrids and their parental stocks was analyzed by RAPD using 10 of the above 38 primers. The average heterozygosity and genetic distance were calculated. The results suggested that the filial generation could inherit a little more genetic materials from paternal fish than that from maternal fish.
ZHANG Xue-kun; CHEN Li; YIN Jia-ming; TANG Zhang-lin; LI Jia-na
14 yellow-seeded rapeseed lines (Brassia napus L. ) from different genetic sources were used toanalyze diversity of testa pigments content, oil and protein content, and RAPD markers. The results showedthat the anthocyanin and melanin were the most important pigments in testa and their content were responsiblefor the variation in seed color ranging from orange to black yellow, 14 yellow-seeded lines could be classifiedinto 3 groups., high anthocyanin content group with anthocyanin content over 2.54 mg g-1 DW, the seed colorwas light yellow or orange; low pigments content group with low content of anthocyanin and melanin, the testawas transparent and the seed color was light yellow, greenish yellow or twany; high melanin content groupwith melanin content over 178.4U(A290nm), the testa was black, the seed color was black yellow. Oil eantentchanged from 36.2% to 45.5%, protein content from 21.1% to 27.7%, and the correlation analysis revealedthat the oil content is highly significantly negatively correlated with the protein content. The cluster analysisshowed that the extensive genetic variation existed among 14 yellow-seeded lines by using unweighted pairedgroup method with arithmetic average (UPGMA) based on RAPD markers which were amplified with decamerprimers, the genetic similarity among them ranged from 0.25 to 0. 909, and 14 yellow-seeded lines could putinto 2 clusters corresponding to genome difference.
El-Batanouny, M. H.; Amin, R. M.; Ibrahium, M. K.; El Gohary, S.; Naga, M. I.; Salama, M. S.
Helicobacter pylori is one of the most common causes of chronic bacterial infections in humans and is important in the pathogenesis of gastrointestinal disease, such as duodenal ulcer, gastric ulcer, Gastric adenocarcinoma, and lymphoma. Gastric adenocarcinoma remains one of the leading causes of cancer death in the world. The objective of this study was to assess the effect of photodynamic treatment and medication treatment of Helicobacter pylori using RAPD-PCR. The lethal photosensitization effect was determined by mixing suspensions of H.pylori with Toluidine blue O (TBO) and plating out on blood agar before irradiation with Helium neon (He-Ne) 632.8 nm. The susceptibility of Helicobacter pylori isolates to metronidazole and azithromycin were examined by E-test. Nine random primers were used to screen genetic polymorphism in DNA of different H.pylori groups. Six of them produced RAPD products while three failed to generate any product. The resulting data showed that, although the overall genetic differences between control groups and laser treated groups was higher than that between control groups and azithromycin treated groups yet it still law genetic variability. The main cause of cell death of PDT using TBO as a photosensitizer was mainly cell wall and cytoplasmic membrane.
Full Text Available RAPD markers (Random Amplified Polymorphic DNA were used for analysing 145 individuals (128 T. grandiflorum and 17 T. bicolor from the ex situ Theobroma genus germplasm bank at Instituto Sinchi, located at San José del Guaviare. 5 primers able to generated polymorphism were selected from an initial set of 20, generating 114 bands that enable to us to distinguish between more than 99% of individuals analysed: 57 bands for T. grandiflorum (84.2% polymorphic, 45 bands for T. bicolor (26.7% polymorphic and 12 bands shared between the two species (58.3% polymorphic. A high degree of intra-specific similarity particularly in T. bicolor was established from the similarity matrix obtained by using the Dice index and represented in a UPGMA dendrogram and the principal components analysis (PCA. The comparison of this analysis with a previous morpho-agronomic evaluation of some T. grandiflorum individuals revealed that the groups generated on the basis of its agronomic and morphological traits were heterogeneous at molecular level. The obtained information will be used as a tool in strategies regarding maintenance, enrichment and use of the germplasm bank. Key words: Theobroma grandiflorum, Theobroma bicolour, RAPD, molecular characterisation.
Full Text Available The genetic variability and phylogenetic relationships in oil palm (Elaeis guineensis Jacq. were studied using RAPD (Random Amplified Polymorphic DNA. Leaf samples of 151 plants were collected from different areas in southern Thailand. DNA from the leaf samples was isolated using CTAB buffer and screened by decamer oligonucleotide primers. Among the total of 160 primers screened, 7 primers (OPB-08, OPR-11, OPT-06, OPT-19, OPAB-01, OPAB-09 and OPAB-14 were chosen to analyse for genetic variation in 151 individuals representing 52 dura, 60 tenera and 39 pisifera. Two hundred and nine amplified fragments were obtained from 7 primers with an average of 29.85 RAPD markers per primer. A dendrogram showing genetic similarities among oil palm was constructed based on polymorphic bands using UPGMA (Unweighted Pair-Group Method Using Arithmetic Average. Cluster analysis was performed using the SPSS program, which revealed four major clusters: 1 dura, tenera and pisifera from Paorong Oil Palm Company, Oil Palm Research Center, dura and tenera from private plantation in Krabi, and dura from Thepa Research Station;2 dura and tenera from Thai Boonthong Company, pisifera and tenera from Thepa Research Station, dura, tenera and pisifera from Klong Hoi Khong Research Station; 3 and 4 dura and tenera from Univanit Company, respectively. In general, a similarity index showed relatively high levels of 0.6 or greater.
Thul, Sanjog T; Darokar, Mahendra P; Shasany, Ajit K; Khanuja, Suman P S
The taxonomic identity of Capsicum species is found to be difficult as it displays variations at morpho-chemical characters. Twenty-two accessions of six Capsicum species, namely, C. annuum, C. baccatum, C. chinense, C. eximium, C. frutescens, and C. luteum were investigated for phenotypic diversity based on flower color and for genetic differences by molecular makers. The genetic cluster analyses of 27 RAPD and eight ISSR primers, respectively, revealed genetic similarities in the ranges of 23-88% and 11-96%. Principal component analysis of the pooled RAPD and ISSR data further supports the genetic similarity and groupings. Different species showed variations in relation to corolla shade of flower. C. annuum accessions formed a single cluster in the molecular analysis as maintaining their flower characteristic. C. chinense accession shared flower features with the accessions of C. frutescens and were found to be closer at genotypic level. C. luteum was found to be rather closer to C. baccatum complex, both phenotypically and genetically. The only accession of C. eximium presenting purple flowers falls apart from the groupings. The floral characteristics and the molecular markers are found to be useful toward the delineation of the species specificity in Capsicum collection and identification of genetic stock. PMID:21861246
Liu Xunjia; Zheng Yonglian; Liu Jilin
The genetic diversity of maize populations Wuxi (W) from Southwest China, BSSS9(B) from America, Mohuangjiu (M) from Mexico, WBMC0 synthesized by W, B, M as main parents,WBMC1 one cycle selected from WBMC0 were evaluated by RAPD molecular marker. The results showed that :(1) Totally 89 fragments (loci) were amplified by 15 10-mar random primers, the proportion of polymorphic loci were W 76. 4%, B 75. 3%, M 79. 8%,WBMC0 85. 4% and WBMC1 92. 1% respectively; (2) The mean gene heterozygosity based on 89 loci was W 0. 285, B 0. 252, M 0. 296, WBMC0 0. 327 and WBMC1 0. 346; (3) The mean genetic distance based on 89 loci were W 0. 2533, B 0. 2246, M 0. 2481, WBMC0 0. 3006 and WBMC1 0. 3119; (4) The genotypic mean numbers amplified by 15 primers were W 9.1, B 7.8, M 8.5, WBMC0 10. 1 and WBMC1 10. All indexes indicated that the synthesized maize population were more polymorphic than the parent populations in DNA level. One cycle selection did not reduce the variation. The new conception of "genotypic diversity" (the number of genotypes in a population) was provided to describe the genetic diversity for any population being equilibrium or unequilibrium in genetics. The principle and technical system were discussed for evaluating genetic variation of recurrent selection population using RAPD molecular marker.
Twenty four wheat varieties/lines were assessed through RAPD for genetic diversity. Of forty primers, thirteen were able to amplify the genomic DNA and yielded 269 polymorphic bands. The percentage of the polymorphic loci was 86.22%. Nei's genetic diversity (h) ranged from 0.248 to 0.393, with an average of 0.330. Shanon's index ranged from 0.382 to 0.567, with an average of 0.487. The proportion of genetic variation among the populations ( Ds) accounted for 28.58 % of the whole genetic diversity. The level of gene flow (Nm) was 1.25. Some specific RAPD bands were also identified, variety C-591, and QM-4531 contain a specific segment of 4.9 kbp. Whereas SARC-1 and PKV-1600 amplified a specific DNA segment with primer A-09. Marvi-2000 contains two specific segments of 3.2 kb and 200 bp amplified with primer B-07. Genetically most similar genotypes were C-591 and Pasban-90 (76%) and most dissimilar genotypes were Rawal-87 and Khirman (36.1%). On the basis of results, 24 wheat varieties under study could be divided into 'two' groups and five clusters 'A' to 'E. (author)
Sheila Recepute Silveira
Full Text Available The RAPD technique associated with restriction digestion of genomic DNA was used to assess the genetic variability within and among nine populations of Coffea arabica, including six progenies belonging to the Sarchimor germplasm, the progeny PR 77054-40-10 (Catuaí Vermelho IAC 81 x Icatu, and two commercial cultivars (IAPAR 59 and Catuaí Vermelho IAC-81. These populations were evaluated using analysis of molecular variance (AMOVA, genetic similarity among progenies, and percentage of polymorphic loci. A total of 99 RAPD markers were evaluated of which 67 (67.67% were polymorphic. AMOVA showed that 38.5% and 61.5% of the genetic variation was distributed among and within populations, respectively. The fixation index (F ST of the genotypes was 0.385. The mean genetic variability estimated within populations ranged from 15.58 (IAPAR 59 to 8.27 (Catuaí Vermelho IAC 81. A distinct level of genetic variability was revealed for each of the coffee progenies and varieties studied. The methodology used in this investigation was useful to determine the genetic variability within and among C. arabica L. populations providing significant information for coffee breeding.
YOU Feng; WANG Wei; XU Dongdong; ZHU Xiangping; NI Jing; WU Zhihao; XU Yongli; WANG Xincheng; ZHANG Peijun
The hybrid between olive flounder Paralichthys olivaceus and stone flounder Kareius bicoloratus was produced by artificial insemination of olive flounder eggs with stone flounder Sperm. Sinistral and dextral are two types of hybrid progeny after metamorphosis. Karyotypes of both hybrid flounders are the same as those of the two parental species. Of the 22 loci examined from 12 allozymes,12 confirmed hybridization of the paternal and matemal loci in hybrids and no difference was found in allozyme patterns of sinistral and dextral hybrid fishes. RAPD patterns of these specimens were also studied with 38 primers selected from 104 tested. Among them, the PCR products of 30 primers showed hybridization of the paternal and maternal bands. Genetic variation between hybrids and their parental stocks was analyzed by RAPD using 10 of the above 38 primers. The average heterozygosity and genetic distance were calculated. The results suggested that the filial generation could inherit a little more genetic materials from paternal fish than that from maternal fish.
Full Text Available 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 construction of the dendrogram. A total of 589 scorable bands were obtained, 93.12% of them were polymorphic. The Nei�s gene diversity (H of two wild populations were more (0.0504 � 0.0670 and 0.0519 � 0.0953 than the captive population (0.0489 � 0.0850. The clustering pattern obtained by UPGMA method emphasized the wild populations were clustered in one clade and captive population was deviated into another clade. This study proved that RAPD analysis has the ability to discriminate L. calcarifer populations. Further molecular studies, comprising a higher number of molecular tools are still required to precisely evaluate the genetic structure of all seabass populations along the Indian coast.
Mandal, Anup; Lal, Kuldeep Kumar; Mohindra, Vindhya; Singh, Rajeev Kumar; Punia, Peyush; Chauhan, U K; Lakra, Wazir Singh
Twenty-seven enzyme systems, six random amplified polymorphic DNA (RAPD) primers, and two microsatellite loci were tested to determine intraspecific divergence in the natural population of the endangered Indian featherback fish, Chitala chitala, for the first time. The 262 samples of C. chitala were collected from six riverine locations in India: the Satluj, Ganga (Ghagra, Bhagirathi, and Brahmaputra), Mahanadi, and Narmada river systems. The analysis revealed population subdivisions, with an F(ST) value from 0.1235 (95% confidence 0.0868-0.1621) for RAPD and a combined F(ST) of 0.0344 (95% confidence 0.0340-0.0350) for microsatellite loci. An analysis of 38 allozyme loci did not reveal any polymorphism in the samples from any of the riverine localities; a possible explanation for this could be that the ancestors of Chitala could have faced a population reduction in prehistoric periods, as low allozyme variation is also reported for other species of Chitala from south Asia. PMID:19184406
Cytological effects and RAPD analysis of root-tip cells of Allium fistulosum L., the dry seeds were irradiated by 12 C6+ heavy ions, were studied. Micronucleus and chromosomal aberration observed in the root-tip cells showed that obvious 'linear rise' trend was in the frequency of micronucleus, multi-micronucleus and total chromosomal aberration. But a 'U- formed' trend was observed only in the chromosomal aberration frequency. At the same time, RAPD analysis revealed 12C6+ heavy ions exerted great effect on DNA variation of Allium fistulosum L.. Flawing bands, increasing bands and the ones with different transport ratios were observed in amplified bands of 28 selected primers. The DNA variation rate for 30, 90 and 180Gy treatments was 29.91%, 41.05% and 22.14%, respectively. This study also found that high micronucleus frequency and chromosomal variation lead to high lethality, resulting in low DNA variation rate in the survived plants which seeds were irradiated with high doses. (authors)
Gillies, A C; Navarro, C; Lowe, A J; Newton, A C; Hernández, M; Wilson, J; Cornelius, J P
Swietenia macrophylla King, a timber species native to tropical America, is threatened by selective logging and deforestation. To quantify genetic diversity within the species and monitor the impact of selective logging, populations were sampled across Mesoamerica, from Mexico to Panama, and analysed for RAPD DNA variation. Ten decamer primers generated 102 polymorphic RAPD bands and pairwise distances were calculated between populations according to Nei, then used to construct a radial neighbour-joining dendrogram and examine intra- and interpopulation variance coefficients, by analysis of molecular variation (AMOVA). Populations from Mexico clustered closely together in the dendrogram and were distinct from the rest of the populations. Those from Belize also clustered closely together. Populations from Panama, Guatemala, Costa Rica, Nicaragua and Honduras, however, did not cluster closely by country but were more widely scattered throughout the dendrogram. This result was also reflected by an autocorrelation analysis of genetic and geographical distance. Genetic diversity estimates indicated that 80% of detected variation was maintained within populations and regression analysis demonstrated that logging significantly decreased population diversity (P = 0.034). This study represents one of the most wide-ranging surveys of molecular variation within a tropical tree species to date. It offers practical information for the future conservation of mahogany and highlights some factors that may have influenced the partitioning of genetic diversity in this species across Mesoamerica. PMID:10651917
YOU Feng; ZHANG Peijun; WANG Keling; XIANG Jianhai
Population genetics of the left-eyed flounder, Paralichthys olivaceus, including natural and cultured stocks distributed in the coastal waters near Qingdao of eastern maritime China, was analyzed in allozyme and RAPD. The results showed that among total 29 gene loci of 15 isozymes, 9 and 7 were polymorphic in natural and cultured stocks, respectively. The status of genetic diversity in P olivaceus is low in terms of polymorphic loci in chi-square test and genetic departure index of Hardy-Weinberg equilibrium. More alleles in IDHP, CAT, GDH and Ldh-C allozymes were found in the fish, which could be used as markers in assortive breeding and distinguishing stock, population or species evolution. Total 88 and 86 RAPD bands ranging from 200 to 2 500 bp were recognized individually in average of 7.8-8.0 bands per primer. The genetic diversity in cultured stock is lower than that in natural ones showing an obviously decreasing genetic divergence. Therefore, effective countermeasures must be taken to protect genetic resources of marine cultured fishes. The 2 markers have their own pros and cons. Combining the 2 markers to investigate the genetic variation of populations is suggested. The results provide basic data of this flounder and they are useful for studying genetic improvement and genetic resources of the fish.
Full Text Available The osmotic stress tolerance of B. japonicum strains assessed according to their persistence in PEG solution. The lowest tolerance to osmotic stress was observed in strain 511 (43.3%, and the highest tolerance was observed for strain D216 (3.3% growth reduction in presence of PEG. PGP traits of B. japonicum strains were tested. None of five B. japonicum strains produced siderophore, strains 511 and 518 had the urease ability, and only B. japonicum 518 strain showed the ability to solubilize insoluble tricalcium phosphate. RAPD analysis, using AP10, BC318, AF14 and SPH1 primers, indicated genetic differences between Bradyrhizobium strains. The first group (strains 3, 6 and 518 showed more than 80% similarity. Strains 511 and D216 formed separate clusters. Difference between strains D216 and the other strains were more than 60%, with maximum value of 72% in comparison with strain 511. Plant-growth promoting (PGP traits, osmotic stress tolerance and RAPD analysis highlighted strain D216 as useful for further investigation of B. japonicum impact on drought reduction in symbiosis with soybean. [Projekat Ministarstva nauke Republike Srbije, br. TR 31022 i br. TR 31072
Maria Cristina Rocha Cordeiro
Full Text Available Actually mango (Mangifera indica, L. is considered one of the largest Brazilian fruitbusiness for the export market. Cultivar selection having high fruit quality is a fundamental step to obtain excellent results in this business. A mango breeding program based on intervarietal hybridization may produce new improved cultivars for mango growers. Mango hybrids have been obtained by controlled or open crosses. In the last one, it is important to identify the male parent because it is useful for the genetic cultivar history, thus it is important for planning further improvements. This work presents a parentage test using among others parameters RAPD (Random amplified Polymorphic DNA markers to estimate the male parent of the selected hybrids in an open cross plot by using five mango cultivars densely planted in a latin square design.Atualmente, o cultivo da mangueira (Mangifera indica, L. é considerado um dos maiores agronegócios frutícolas brasileiros para o mercado externo. A seleção de cultivares com elevada qualidade de frutos é uma etapa fundamental para o sucesso deste negócio. O melhoramento genético da mangueira, baseado em hibridações intervarietais, pode originar cultivares superiores aos padrões disponíveis no mercado, ofertando uma nova alternativa ao produtor. Híbridos de mangueira têm sido obtidos por meio de cruzamentos controlados ou abertos. Neste último, é importante identificar o genitor masculino visando a obter a história genética da cultivar além de ser importante no planejamento do melhoramento subseqüente. Este trabalho apresenta um teste de paternidade, utilizando, entre outros parâmetros, marcadores RAPD (Random Amplified Polymorphic DNA, para estimar o genitor masculino de híbridos selecionados em uma área de cruzamento aberto contendo cinco cultivares de manga plantadas em alta densidade, no delineamento experimental do quadrado latino.
Random amplified polymorphic DNA analysis of effective Rhizobium sp. associated with beans cultivated in brazilian cerrado soils Caracterização, via RAPD, de estirpes efetivas de Rhizobium sp associadas ao feijoeiro cultivado em solos de cerrado no Brasil
Isnia Aparecida de Oliveira
Full Text Available Efficient bean nodulating Rhizobium strains, isolated from different Brazilian cerrado soils, were characterized by RAPD. This study showed great genetic heterogeneity among R. tropici and R. leguminosarum bv. phaseoli strains and allowed the constitution of genetic clusters, besides indicating the most suitable primers for this characterization. The groups of genetically distinct strains can be used in competitiveness studies to select appropriate Rhizobium strains for bean inoculation in cerrado soils.Estirpes de Rhizobium eficientes na nodulação do feijoeiro, isoladas de diferentes solos da região do cultivo dessa leguminosa nos cerrados brasileiros, foram caracterizadas via RAPD. Esse estudo mostrou grande heterogeneidade genética entre as estirpes de R. tropici e R. leguminosarum bv. phaseoli testadas e permitiu a definição de grupos genéticos, além de indicar os "primers" mais adequados para essa caracterização. Os grupos de estirpes geneticamente distintas podem ser usados em estudos de competitividade, importantes para obtenção de resultados positivos na inoculação dessa leguminosa em solos de cerrado.
Biswajit Maiti; Malathi Shekar; Rekha Khushiramani; Iddya Karunasagar; Indrani Karunasagar
Sixty five isolates of Vibrio harveyi were subjected to random amplified polymorphic DNA (RAPD)-PCR analysis and protein profiling to investigate the genetic variability among V. harveyi prevalent along the coast and also assess the discriminating ability of these two molecular methods. A total of 10 RAPD primers were assayed for their specificity in detecting V. harveyi, of which only two primers: PM3 and CRA25 were highly reproducible and found suitable for use in RAPD-PCR. The genetic diversity among V. harveyi isolates assessed by RAPD-PCR using PM3 primer yielded 35 different RAPD patterns which clustered the isolates into 15 groups at 72% similarity level. Similarly, RAPD-PCR with CRA25 clustered the 38 patterns into 10 groups at 74% similarity. The discriminatory index $(D)$ value calculated for RAPD fingerprints generated with PM3 and CRA25 were 0.90 and 0.85, respectively. On the other hand, molecular typing of V. harveyi using whole cell proteins generated profiles that showed no major difference indicating the technique to be not useful in typing strains of this bacterium. However, a few of the isolates showed the presence of unique band of 28 kDa that needs to be further investigated to understand the role of the protein in disease process if any.
João Batista Tavares da Silva
Full Text Available The objective of this work was to study the genetic variability of the grasshopper Rhammatocerus schistocercoides (Orthoptera: Acrididae using RAPD analysis among individuals from three populations, one from Colombia and two from Brazil (Goiás and Mato Grosso States. Ninety scorable binary markers were obtained by fingerprinting with 11 oligonucleotide primers. Most of the polymorphism was attributed to 42 markers with variable frequency among the different populations. Although the existence of significant difference among populations (PO objetivo deste trabalho foi estudar a variabilidade genética do gafanhoto Rhammatocerus schistocercoides (Orthoptera: Acrididae por meio da análise de RAPD entre indivíduos de três populações, uma da Colômbia e duas do Brasil (Goiás e Mato Grosso. Noventa marcadores binários foram selecionados através de análise de polimorfismo com o uso de 11 oligonucleotídeos. A maior parte do polimorfismo observado foi atribuída a 42 marcadores com freqüência variável entre as diferentes populações. Apesar da existência de diferença significativa interpopulacional (P<0,0001, grande proporção da variabilidade genética foi detectada dentro das populações (87,7% da variação total. As distâncias entre as populações colombianas e brasileiras foram 0,12 (P<0,0001 e 0,18 (P<0,0001 para Goiás e Mato Grosso, respectivamente. A distância obtida entre Goiás e Mato Grosso foi 0,06 (P<0,0001. Estes dados indicam que as diferenças fenotípicas entre populações estão associadas principalmente às distâncias geográficas entre as populações do Brasil e a da Colômbia.
SAWAZAKI HAIKO ENOK
Full Text Available Estudou-se a variabilidade genética em couve (Brassica oleracea L. var. acephala D.C. tipo manteiga por intermédio do polimorfismo enzimático em gel de poliacrilamida e do polimorfismo de DNA, denominado RAPD (Random Amplified Polymorphic DNA, com base na amplificação de segmentos de DNA ao acaso. Avaliaram-se quinze clones de couve-manteiga do Banco Ativo de Germoplasma do Instituto Agronômico (IAC, utilizando-se extratos de folhas para análise de isoenzimas e marcador RAPD com os "primers" dos kits A e B da Operon Technologies. Entre as isoenzimas estudadas, as mais polimórficas foram as fosfoglucomutase (PGM, peroxidase (PRX e esterase (EST, tendo o sistema PGM realizado a melhor caracterização. Verificou-se a ocorrência de variabilidade genética por meio de isoenzimas e RAPD, porém não foi observada a similaridade entre os dendrogramas obtidos por ambos os tipos de marcadores, sugerindo que as isoenzimas forneceram menos informação sobre o genoma. A maior eficácia do RAPD foi devida à possibilidade de processar maior número de análises, evidenciando mais detalhes sobre o genoma.
Justesen, Annemarie Fejer; Yohalem, David; Bay, Anne; Nicolaisen, Mogens
DNA sequence analysis of the internal transcribed spacer region 1 (ITS1) and random amplified polymorphic DNA (RAPD) markers were used to survey genetic variability in relation to agronomic and regional factors among 60 isolates of Thanatephorus cucumeris (anamorph Rhizoctonia solani) collected from lesions on potato stems or sclerotia of potato tubers. Based on comparative sequence analysis it was shown that all isolates belonged to anastomosis group 3 subgroup Potato Type (AG-3 PT). ITS1 sequence polymorphisms were found within 45 of the 60 isolates showing that different types of the ITS-region are present in individual isolates. Cloning and sequence analysis of the ITS1 region from three selected isolates with sequence polymorphism showed that two different ITS1-types were present in each isolate. RAPD analysis identified 51 RAPD-phenotypes among the 60 investigated isolates indicating a high level of diversity within the subgroup AG-3 PT. Putative clonal isolates with identical RAPD- and ITS1-types were identified within fields, and in one case the same phenotype was found in two different fields separated by several hundred kilometers. Population subdivision analysis based on phenotypic as well as genotypic diversities showed differentiation among populations from different fields when isolates were sampled from tubers, indicating restricted gene flow among soil populations. Low differentiation was seen among field populations sampled from stems, indicating that gene flow is taking place. The population structure was not influenced by the previous crop in the rotation nor by the two cultivars 'Sava' and 'Bintje'. PMID:15000234
Ruanet, V V; Kochieva, E Z; Ryzhova, N N
The results of studies based on multilocus molecular analyses, including random amplified polymorphic DNA (RAPD), inter-simple sequence repeat (ISSR), and amplified fragment length polymorphism (AFLP) analyses, are usually presented in the form of images (electrophoregrams, photographs, etc.). The interpretation of this information is complicated, labor-consuming, and subjective. Artificial neural networks (ANNs), which are ideal "image processors," may be useful when solving such tasks. The possibility of using ANNs for the treatment of the results of RAPD and ISSR analyses has been studied. The RAPD and ISSR spectra have been studied in fragments of DNA of plants from the genus Capsicum L. (peppers). The results of clustering the accessions studied by means of the unweighted pair-group method with arithmetic averages (UPGMA), which is often used for phylogenetic constructions based on RAPD and ISSR data, serve as expert estimates. Fundamentally new methods of genetic polymorphism estimation using ANN technologies, namely, self-organizing feature maps (SOFMs) have been developed. The results show that the clusters obtained with the use of UPGMA and SOFM coincide by more than 90%; taking into account that ANNs can deal with high noise levels and incomplete or contradictory data, the approach proposed may prove to be efficient. PMID:15810617
Full Text Available Bemisia tabaci (Genn. was considered a secondary pest in Brazil until 1990, despite being an efficient geminivirus vector in beans and soybean. In 1991, a new biotype, known as B. tabaci B biotype (=B. argentifolii was detected attacking weed plants and causing phytotoxic problems in Cucurbitaceae. Nowadays, B. tabaci is considered one of the most damaging whitefly pests in agricultural systems worldwide that transmits more than 60 different plant viruses. Little is known about the genetic variability of these populations in Brazil. Knowledge of the genetic variation within whitefly populations is necessary for their efficient control and management. The objectives of the present study were to use RAPD markers (1 to estimate the genetic diversity of B. tabaci populations, (2 to study the genetic relationships among B. tabaci biotypes and two other whitefly species and (3 to discriminate between B. tabaci biotypes. A sample of 109 B. tabaci female individuals obtained from 12 populations in Brazil were analyzed and compared to the A biotype from Arizona (USA and B biotype from California (USA and Paraguay. Trialeurodes vaporariorum and Aleurodicus cocois samples were also included. A total of 72 markers were generated by five RAPD primers and used in the analysis. All primers produced RAPD patterns that clearly distinguished the Bemisia biotypes and the two other whitefly species. Results also showed that populations of the B biotype have considerable genetic variability. An average Jaccard similarity of 0.73 was observed among the B biotype individuals analyzed. Cluster analysis demonstrated that, in general, Brazilian biotype B individuals are scattered independently in the localities where samples were collected. Nevertheless, some clusters were evident, joining individuals according to the host plants. AMOVA showed that most of the total genetic variation is found within populations (56.70%, but a significant portion of the variation is found
@@The somaclonal mutant HX_3 has shown a broad spectrum resistance to bacterial blight. To study the inheritance of the bacterial blight resistance in HX_3, a cross was made between HX_3 and a susceptible cultivar Longtefu A. The F2 population of 418 plants was inoculated with Chinese bacterial blight strain Zhe 173 (pathotype Ⅳ ). Results showed that the F2 progenies segregated in a ratio of 3R∶ 1S (324 resistant plants and 94 susceptible plants). From the plants tested, 114 individuals (86 resistant and 28 susceptible) were chosen randomly for RAPD analysis. Twelve highly resistant and 12 highly susceptible plants were selected to form a resistant pool and a susceptible pool, respectively.
Singh, N; Lal, R K; Shasany, A K
Plantago ovata, popularly known as isabgol, has great commercial and medicinal importance due to thin rosy white membranous seed husk. Isabgol seeds and husks have emollient, demulcent and laxative properties. We used both biometric and molecular techniques to assess the genetic variability and relatedness of 80 germplasm accessions of Plantago spp (P. ovata, P. lanceolata, and P. major) collected both from India and abroad. The range of D2 values (2.01-4890.73) indicated a very high degree of divergence among the accessions. Based on the degree of divergence, 80 accessions/genotypes were grouped into seven clusters. Thirty-six accessions were analyzed through RAPD profiling for similarity and genetic distances, using 20 random primers. Intraspecific differences in all three species were smaller [range for P. ovata (2-17%), P. lanceolata (3-15%), P. major (2-11%)] than interspecific diversity. These highly divergent lines could be used to produce superior hybrids. PMID:19876869
Bananas and plantains are one of the most important fruit crops and a staple food for millions in the tropical and subtropical countries. Owing to the constraints of vegetative propagation, sterility and triploid nature of most of the cultivars, mutation breeding and biotechnological methods are useful tools for developing cultivars resistant to diseases and pests. Herein, multiple shoot cultures of banana Giant Cavendish (Musa spp. AAA group) were gamma-irradiated (5, 10 and 30 Gy). Field evaluation of these exhibited variation for several agronomic traits including dwarf stature and early flowering. The selections are now being studied for stability. Molecular analysis using RAPD markers indicated polymorphism and some of the dwarf selections were analyzed using dwarf-specific SCAR marker confirming their dwarf behavior. The results suggest that the gamma irradiation is useful for the isolation of agronomic variants in Cavendish bananas (author)
YU Shun-wu; CHEN Bao-tang; TAO Ai-lin; ZHANG Duan-pin
To transfer desirable resistance traits from O. minuta to O. sativa, intergeneric hybrid plants between O. sativa (AA, 2n=2X=24) and O. minuta (BBCC, 2n=4X=48) were produced by embryo rescue after sexual cross. Morphological observation and chromosome counts indicated their hybrid status (ABC, 2n =3X=36). Genomic in situ hybridization (GISH) was further applied to confirm the parentage of the chromosomes of F1 hybrids. Chromosomes of O. minuta and O. sativa were distinguishable in the hybrids in different fluorescence colors. GISH indicated that A and BC chromosomes were not randomly assembled in a cell.RAPD profiles unequivocally revealed their hybrids with double parent patterns. The results of blast tests showed that the hybrids had obtained disease resistance from O. minuta, and had a level of susceptibility between the parents.
Degen, B; Caron, H; Bandou, E; Maggia, L; Chevallier, M H; Leveau, A; Kremer, A
The fine-scale spatial genetic structure of eight tropical tree species (Chrysophyllum sanguinolentum, Carapa procera, Dicorynia guianensis, Eperua grandiflora, Moronobea coccinea, Symphonia globulifera, Virola michelii, Vouacapoua americana) was studied in populations that were part of a silvicultural trial in French Guiana. The species analysed have different spatial distribution, sexual system, pollen and seed dispersal agents, flowering phenology and environmental demands. The spatial position of trees and a RAPD data set for each species were combined using a multivariate genetic distance method to estimate spatial genetic structure. A significant spatial genetic structure was found for four of the eight species. In contrast to most observations in temperate forests, where spatial structure is not usually detected at distances greater than 50 m, significant genetic structure was found at distances up to 300 m. The relationships between spatial genetic structure and life history characteristics are discussed. PMID:11737299
卢钢; 陈杭; 等
A genetic linkage map comprised of 131 loci was constructed with an F2 population derived from an inter-subspecific cross between Brassica campestris L.ssp.chinensis cv.aijiaohang” and ssp.rapifera cv.,”'isihai”.The genetic map included 93 RAPD loci,36 AFLP loci and 2 morphological loci organized into 10 main linkage groups(LGs) and 2 small groups,covering 1810.9cM with average distance between adjacent markers being approximately 13.8cM.The map is suitable for identification of molecular markers linked to important agronomic traits.QTL analysis,and even for marker-assisted selection in breeding programs of Chinese cabbage and turnip.
Callejas, C; Ochando, M D
Random amplified polymorphic DNA (RAPD) markers were used to estimate the population structure and phylogenetic relationships among the eight species of the genus Barbus that inhabit the Iberian Peninsula. Ten random oligodecamers were used to amplify DNA from 232 fish sampled from 15 populations. A total of 270 markers were detected that revealed low levels of genetic variability. The conclusions of cluster analysis indicate two main branches and three well-differentiated groups: north-eastern, Mediterranean and Atlantic. This clustering mainly reflects the evolutionary history of the genus, which is closely related to the paleogeography of the Iberian Peninsula. The contribution and application of these results to the conservation of the species, to their taxonomic status and to the process of colonization of the Iberian Peninsula by the genus Barbus are discussed. PMID:12080368
Full Text Available Many factors have contributed to the destruction of fish habitats. Hydroelectric dams, water pollution and other environmental changes have resulted in the eradication of natural stocks. The aim of this study was to detect the genetic variation in Prochilodus marggravii from three collection sites in the area of influence of the Três Marias dam (MG on the São Francisco river (Brazil, using the RAPD technique. The results obtained revealed that the fish in the downstream region nearest the dam have a higher similarity coefficient than those from the other sampling sites that may be related to differences in environmental characteristics in these regions. Additionaly, significant differences in the band frequencies were observed from one collection site to another. These both findings suggest the occurrence of a structured population and have important implications for the conservation of the genetic variability of distinct natural P. marggravii stocks.
Sharma, Sweta K.
Jatropha curcas (Euphorbiaceae), a drought resistant non edible oil yielding plant, has acquired significant importance as an alternative renewable energy source. Low and inconsistent yields found in field plantations prompted for identification of high yielding clones and their large scale multiplication by vegetative propagation to obtain true to type plants. In the current investigation plantlets of J. curcas generated by axillary bud proliferation (micropropagation) using nodal segments obtained from selected high yielding genotypes were assessed for their genetic stability using Randomly Amplified Polymorphic DNA (RAPD) and Amplified Fragment Length Polymorphism (AFLP) analyses. For RAPD analysis, 21 out of 52 arbitrary decamer primers screened gave clear reproducible bands. In the micropropagated plantlets obtained from the 2nd sub-culture, 4 out of a total of 177 bands scored were polymorphic, but in the 8th and 16th sub-cultures (culture cycle) no polymorphisms were detected. AFLP analysis revealed 0.63%, 0% and 0% polymorphism in the 2nd, 8th and 16th generations, respectively. When different genotypes, viz. IC 56557 16, IC 56557 34 and IC 56557 13, were assessed by AFLP, 0%, 0.31% and 0.47% polymorphisms were found, respectively, indicating a difference in genetic stability among the different genotypes. To the best of our knowledge this is the first report on assessment of genetic stability of micropropagated plantlets in J. curcas and suggests that axillary shoot proliferation can safely be used as an efficient micropropagation method for mass propagation of J. curcas. © 2011 Elsevier B.V.
Full Text Available Background: The population size of plants affects on population genetic variation. Materials and Methods: We studied the impact of population size on genetic variation in populations of Capparis spinosa (caper, Capparaceae using RAPDs in East Azerbaijan (Iran. Within-population genetic diversity was estimated based on Nei`s and Shanonn`s diversity using Popgen, and genetic similarity among the populations was studied from a UPGMA dendrogram based the matrix of Nei’s distances obtained through SHAN. Difference in the level genetic variation between small-sized and large-sized populations was tested using Mann-Whitney U test, and correlation between geographical and genetic distances among populations was examined by Pearson test (SPSS, 11.3. Total genetic variation was partitioned into within and among populations based on AMOVA using Arlequin. Results: The polymorphism levels of RAPDs bands among the populations ranged from 48.8% to 81.4%, and within-population Nei’s diversity varied from 0.1667 to 0.2630. Genetic variation in small-sized populations (0.1667 to 0.1809 was significantly lower than the variations in large-sized populations (0.2158 -0.2630 (N= 7, P0.674, Pearson correlation test. Conclusions: Population size has a dramatic impact on its genetic diversity. The results revealed that fragmentation of caper population in the study region has most likely occurred recently. The low genetic diversity revealed within caper populations indicates high risk of extinction and suggests that urgent conservation action is needed to recover diversity in these populations.
The aim of this study is to examine the discriminatory power of random amplified polymorphic DNA (RAPD) marker in Jatropha curcas, and to determine the effect of various dose exposures (0, 5, 10, f, 20 and 25 Kr) of gamma rays on J. curcas, at molecular level. All the ten random primers used produced reproducible polymorphic bands. PCR products of mutant genome revealed a total of 40 bands, out of which 27 were polymorphic. Polymorphism information content (PIC) values were ranged from 0.00 to 0.40 and the highest PIC value of 0.40 was observed in primer OPU-13 followed by primers OPAL-II and OPT-18 (0.30) while no PIC value were reported in primers OPH-18 and OPM-13. Jaccard's coefficient of similarity varied from 0.476 to 0.723, indicative of high level of genetic variation among the mutants studied. UPGMA cluster analysis indicated three distinct clusters, one comprising control while the second included four mutants viz., 10, 15, 25 and 20 Kr. The mutant 5 Kr remained distinct and formed third cluster indicating its higher genetic diversity from the rest of the mutants and control. The primer OPU-13 produced maximum number of bands (8) showed highest discriminatory power and PIC (0.40) by showing maximum number of polymorphic bands (5) when compared to other primers used. The study reveals that RAPD molecular markers can be used to assess polymorphism among the mutants and can be a useful tool to supplement the distinctness, uniformity and stability analysis for plant varietal identification and protection. (author)
Asensio, Luis; González, Isabel; Fernández, Alicia; Rodríguez, Miguel A; Lobo, Esther; Hernández, Pablo E; García, Teresa; Martín, Rosario
A random amplified polymorphic DNA (RAPD) method was developed for the specific identification of grouper (Epinephelus guaza), wreck fish (Polyprion americanus), and Nile perch (Lates niloticus) fillets. Using two different reaction primers (S1 and L1), RAPD analysis produced clear fingerprints from which the three fish species could be easily identified. This approach is rapid and reliable and offers the potential to detect fraudulent or unintentional mislabeling of these species in routine seafood authentication analysis. PMID:11848581
Cláudia Pombo Sudré
Full Text Available The genus Dimorphandra has distinguish relevance considering either medicinal or biodiversity aspects because it includes two species that are economically important flavonoids sources for pharmachemical industry (D. mollis Benth. and D. gardneriana Tul., and species endemic to Brazil, such as D. jorgei Silva and D. wilsonii Rizz., threatened by extinction. In order to evaluate variability among accessions of D. mollis (fava-d'anta, D. gardneriana and D. wilsonii, it was collected fruit from individual plants from three Brazilian states in a total of 57 accessions, which were analyzed with RAPD markers. It was used 20 seeds per progeny; the DNA was extracted from fully-formed young leaves, which were collected in bulk. The data were analyzed using a binary matrix, in which the score one represented presence of a band and zero, absence. The similarity matrix was developed by using the arithmetic complement of the Jaccard index, later grouped based on the Neighbor Joining algorithm. It was found considerable intra and inter specific variability in Dimorphandra spp., which were separated into four groups. Though genetic variability was found, the collecting trips showed that most of these areas are subject to loss of genetic resources of fava-d'anta due to the following factors: continuous anthropic activity, propensity for natural fires, and loss of natural seed dispersers (large fruit-eating mammals. Therefore, protections of these areas and ex situ conservation are essentials for the maintenance of genetic variability of these species.O gênero Dimorphandra tem grande relevância, sobretudo nos aspectos medicinais e de biodiversidade, por incluir duas espécies que são importantes economicamente como fontes de flavonoides para indústria farmacoquímica (D. mollis Benth. e D. gardneriana Tull., e espécies endêmicas do Brasil, como a D. jorgei Silva e D. wilsonii Rizz., sendo esta ameaçada de extinção. Objetivando avaliar a variabilidade
Andréa Alves do Egito
Full Text Available 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 indicated that the greatest portion of detected variability was due to differences between individuals within populations (75.47%. Analysis of the genetic variability between pairs of populations presented higher estimates for the five Pantaneiro populations with the Arabian breed, while lowest estimates were presented by pairs formed among the Pantaneiro populations with the Mangalarga Marchador. Highest genic diversity was shown by the Pantaneiro (0.3396, which also showed highest genetic distance with the Arabian and lowest with Mangalarga Marchador breed. UPGMA dendrogram showed distinct differences between naturalized (Pantaneiro and Mangalarga Marchador and exotic (Arabian and Thoroughbred breeds. In the dendrogram generated by UPGMA method, the similarity matrix generated by the Jaccard coefficient showed distinction between the naturalised breeds, Pantaneiro and Mangalarga Marchador, and the exotic breeds, Árab and English Thoroughbred. Results suggest that the Pantaneiro presents a higher genetic variability than the other studied breeds and has a close relationship with the Mangalarga Marchador.Amostras de sangue foram coletadas de cavalos Pantaneiros de cinco regiões dos estados de Mato Grosso do Sul e Mato Grosso. As raças Mangalarga Marchador, Árabe e Puro-Sangue Inglês (PSI usando marcadores moleculares RAPD-PCR (Random Amplified Polymorphic DNA - Polymerase Chain Reaction foram incluídas no intuito de se calcular as distâncias gen
Maria Selva Senor
Full Text Available En el presente articulo se reconstruyen las trayectorias matrimoniales de dos familias de la élite en el Río de la Plata entre 1776 y 1810 con el objeto de descubrir las estrategias seguidas por las mismas y las repercusiones del azar. Desde una perspectiva microanalitica y a partir de la utilización de la noción de redes sociales se intentará ilustrar acerca de las reglas sociales en una sociedad con variados contextos normativos superpuestos, que incluyen asimismo a la política del Estado y en la que los agentes pueden manipular reglas y tomar decisiones haciendo que el grado de incertidumbre sea mayor.
Ganguly, Swagata; Saha, Pabitra; Chatterjee, Moytrey; Roy, Surajit; Ghosh, Tamal Kanti; Guha, Subhasish K.; Kundu, Pratip K.; Bera, Dilip K.; Basu, Nandita; Maji, Ardhendu K.
Post Kala-azar Dermal Leishmaniasis (PKDL) is a chronic but not life-threatening disease; patients generally do not demand treatment, deserve much more attention because PKDL is highly relevant in the context of Visceral Leishmaniasis (VL) elimination. There is no standard guideline for diagnosis and treatment for PKDL. A species-specific PCR on slit skin smear demonstrated a sensitivity of 93.8%, but it has not been applied for routine diagnostic purpose. The study was conducted to determine the actual disease burden in an endemic area of Malda district, West Bengal, comparison of the three diagnostic tools for PKDL case detection and pattern of lesion regression after treatment. The prevalence of PKDL was determined by active surveillance and confirmed by PCR based diagnosis. Patients were treated with either sodium stibogluconate (SSG) or oral miltefosine and followed up for two years to observe lesion regression period. Twenty six PKDL cases were detected with a prevalence rate of 27.5% among the antileishmanial antibody positive cases. Among three diagnostic methods used, PCR is highly sensitive (88.46%) for case confirmation. In majority of the cases skin lesions persisted after treatment completion which gradually disappeared during 6–12 months post treatment period. Reappearance of lesions noted in two cases after 1.5 years of miltefosine treatment. A significant number of PKDL patients would remain undiagnosed without active mass surveys. Such surveys are required in other endemic areas to attain the ultimate goal of eliminating Kala-azar. PCR-based method is helpful in confirming diagnosis of PKDL, referral laboratory at district or state level can achieve it. So a well-designed study with higher number of samples is essential to establish when/whether PKDL patients are free from parasite after treatment and to determine which PKDL patients need treatment for longer period. PMID:26485704
Full Text Available Post Kala-azar Dermal Leishmaniasis (PKDL is a chronic but not life-threatening disease; patients generally do not demand treatment, deserve much more attention because PKDL is highly relevant in the context of Visceral Leishmaniasis (VL elimination. There is no standard guideline for diagnosis and treatment for PKDL. A species-specific PCR on slit skin smear demonstrated a sensitivity of 93.8%, but it has not been applied for routine diagnostic purpose. The study was conducted to determine the actual disease burden in an endemic area of Malda district, West Bengal, comparison of the three diagnostic tools for PKDL case detection and pattern of lesion regression after treatment. The prevalence of PKDL was determined by active surveillance and confirmed by PCR based diagnosis. Patients were treated with either sodium stibogluconate (SSG or oral miltefosine and followed up for two years to observe lesion regression period. Twenty six PKDL cases were detected with a prevalence rate of 27.5% among the antileishmanial antibody positive cases. Among three diagnostic methods used, PCR is highly sensitive (88.46% for case confirmation. In majority of the cases skin lesions persisted after treatment completion which gradually disappeared during 6-12 months post treatment period. Reappearance of lesions noted in two cases after 1.5 years of miltefosine treatment. A significant number of PKDL patients would remain undiagnosed without active mass surveys. Such surveys are required in other endemic areas to attain the ultimate goal of eliminating Kala-azar. PCR-based method is helpful in confirming diagnosis of PKDL, referral laboratory at district or state level can achieve it. So a well-designed study with higher number of samples is essential to establish when/whether PKDL patients are free from parasite after treatment and to determine which PKDL patients need treatment for longer period.
A retrospective study of intravenous sodium stibogluconate alone and in combinations with allopurinol, rifampicin, and an immunomodulator in the treatment of Indian post-kala-azar dermal leishmaniasis
Full Text Available Background and Aims: A retrospective analysis of treatment outcome using recommended dose of sodium stibogluconate (SSG alone and in combination with other antileishmanial drugs in adults with post-kala-azar dermal leishmaniasis (PKDL attending as outpatients. Methods: A total of 61 patients seen over ten years were included in the report. All had polymorphic lesions. Diagnosis was based on clinical picture, hailing from kala-azar (KA endemic area, exclusion of other dermatoses, histopathology, and therapeutic response. Patients were distributed into two groups: Group I (n = 32, where SSG was given intravenously; in Group II (n = 29, they were allocated to one of four categories using SSG in combination with other drugs. In the first category, SSG was given along with allopurinol (n = 10; in second with rifampicin (n = 6; and in third with both allopurinol and rifampicin (n = 5. In the fourth category, SSG was administered with an immunomodulator (n = 8, Mw vaccine, known to enhance host Th1 response. Results: Only 12 out of 61 patients completed treatment till histopathologic evidence of cure, five in Group I and seven in Group II, no patient being from third category. None had taken SSG without interruptions. Time taken for papulonodules to subside was similar in both groups, but erythema and induration subsided earlier in Group II. Group I patients attained cure after 120 injections while in Group II it took 95 injections in SSG + allopurinol and Mw vaccine categories respectively, and 110 with SSG + rifampicin. Nevertheless this was insufficient to facilitate compliance. Poor performance and high dropouts related to long duration of therapy, thrombophlebitis, difficulty in accessing veins, disabling rheumatic side-effects and practical problems. Liver, renal and pancreatic functions and ECG remained normal. Conclusion: No major advantage was obtained using allopurinol, rifampicin or Mw vaccine along with SSG as compared to SSG alone.
Palacio M. Juan Diego
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Mediante RAPD's se analizaron 120 muestras foliares de 12 biotipos de Guadua angustifolia Kunth clasificados morfológicamente, procedentes de la cuenca del río Cauca, en el departamento del Cauca, Colombia, para determinar diversidad genética. El ADN se extrajo mediante el protocolo modificado de Dellaporta (1983. Se emplearon los cebadores; OPF-12, OPG-19, OPN-19 y OPP-16 con mayor número de bandas polimórficas. El índice de Shannon (HT = 0.4556 ± 0.1849 señaló diversidad genética total alta y diversidad entre los biotipos y al interior de ellos. El Índice de estructura genética (Gst = 0.5200 e Indice de migración efectiva (Nm = 0.4615 definieron biotipos bien diferenciados. El análisis de similaridad conformó tres grupos a un coeficiente de 0.64. El grupo G1 incluyó los biotipos Curvado, Rayada frecuente, Amarilla Playón, Rayada ancha, Rayada escasa, Convexa, Amarilla, Hembra, Verde irregular y algunos individuos de verde alta. El grupo G2, Verde alta y Macho. El grupo G3, Rayada negra. El estudio molecular agrupó los individuos de forma similar al estudio morfológico, con excepción de los individuos del biotipo Hembra.
Palabras claves: Guadua angustifolia, caracterización molecular, variación genética.
Uso do Random Amplified Polymorphic DNA (RAPD no estudo populacional do Triatoma brasiliensis Neiva, 1911 Use of Random Amplified Polymorphic DNA (RAPD in the populational study of Triatoma brasiliensis Neiva, 1911
Érika C. Borges
Full Text Available Para o estudo de variabilidade genética em Triatoma brasiliensis, o principal vetor da doença de Chagas no Nordeste brasileiro, espécimes de três diferentes populações intradomiciliares foram analisados. Regiões do DNA genômico foram amplificadas utilizando dois iniciadores randômicos através da técnica de RAPD (Random Amplified Polymorphic DNA, visualizados em géis de poliacrilamida corados pela prata. Os perfis originados se mostraram bastante homogêneos quando comparados intrapopulacionalmente. Populações capturadas em duas regiões diferentes do Estado do Ceará também apresentaram homogeneidade entre si, mas, quando comparadas com a população proveniente do Piauí, foi possível diferenciá-las. Esses resultados, preliminares, indicam que o RAPD pode ser usado com sucesso nos estudos de variabilidade em triatomíneos, bem como sugerem a existência de variabilidade entre diferentes populações de T. brasiliensis pertencentes a uma mesma subespécie.We evaluated the genetic variability of Triatoma brasiliensis, the main vector of Chagas disease in Northeast Brazil, using specimens from three populations. Regions of genomic DNA were amplified by RAPD (Random Amplified Polimorphic DNA, using two primers. The products were visualized after polyacrylamide gel electrophoresis followed by silver staining. A dendrogram constructed through the Dice similarity coefficient allowed for separation of the tested specimens into three distinct groups. The populations captured in areas from Ceará State showed similar profiles, but different from that captured in Piauí State. Our results indicate that RAPD can be used successfully in triatomine studies and suggest the presence of genetic variability between different populations of T. brasiliensis.
Roberto Pedroso de Oliveira; Mariângela Cristofani; Marcos Antônio Machado
The objective of this work was to construct linkage maps of 'Pêra' sweet orange [Citrus sinensis (L.) Osbeck] and 'Cravo' mandarin (Citrus reticulata Blanco) using RAPD markers and the pseudo-testcross strategy. The parents were chosen according to the resistance/susceptibility to citrus variegate chlorosis (CVC). The segregation of 176 markers was analyzed in 94 progeny of F1 hybrids, which were obtained from controlled crossings. The linkage map of 'Pêra' sweet orange had 117 markers define...
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
Full Text Available 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 análise de agrupamento evidenciou que não há formação de grupos por área de coleta. Portanto, há pequena variabilidade entre os materiais e esta variabilidade encontra-se distribuída entre as regiões amostradas.Chenopodium ambrosioides L. is known in many parts of Brazil for its medicinal properties, mainly used to control intestinal worms. Its genetic diversity is little studied. The objective of this work was to evaluate the genetic variability of 16 accessions of C. ambrosioides from the cocoa region of Bahia State, Brazil, by the RAPD technique (Random Amplified Polymorphic DNA. Only 6.9% of the 216 amplified RAPD bands were polymorphic and the pattern of dispersion of individuals showed no clustering related to sample site. Therefore, there is low variability among accessions and it is distributed among the accessions from the entire sampled region.
A large numbers of samples of wild soybean accessions and cultivated soybean landraces from various areas in China were analyzed by isozyrme, cytoplasmic DNA RFLP and nuclear DNA RAPD markers in order to reveal their genetic diversity. Greater comprehensive genetic diversity was detected in wild soybean than in cultivated soybean. The genetic plentifulness and the genetic dispersion of wild soybean were 180 (95. 2%) and 0. 2891 while those of cultivated soybean were 154(81.5%) and 0. 2091,respectively. On the most loci, especially on isozyme loci Idh1, Aph, Idh2,and Dia, cytoplasmic DNA RFLP loci cp Ⅰ , cp Ⅲ, mt Ⅳ a and mt Ⅳ b, and nuclear RAPD loci OPAP4-8, OPAP5-1, OPAP9-8 and OPAP20-8, the wild soybeans djffered remarkably from the cultivated ones in allele frequency. These markers could be used in further study on the evolution and origin of the cultivated soybean.
Full Text Available In this study, the HAT-RAPD technique was used to detect DNA methylation in the four plant species, rice (Oryza sativa, petunia (Petunia hybrida, spinach (Spinacea oleracea L. and longan (Dimocarpus longan Lour., which were treated using 5-azacytidine, potassium chlorate (KClO3 and a low temperature induction. Rice and petunia were chosen to be induced because in these species it is known that 5-azacytidine is able to induce hypomethylation in their genomes leading to morphological changes, in particular floral induction, in the developing plants. Using the HAT-RAPD technique, DNA methylation was detected using the restriction enzymes HpaII and MspI in rice and petunia (as expected and in spinach, but was found to be absent in longan, which suggests that floral induction in longan is undetectable in longan.
Full Text Available The present investigation was carried out for the determination of genetic conformity of micropropagated plants of Zingiberofficinale cv Majhauley using Random amplified polymorphic DNA (RAPD technique.Total fifteen different arbitrarydecamers were used as primers to amplify DNA from in vitro plant material to assess the genetic fidelity of 18 months old invitro cultured plantlets. Total 38 amplified reproducible bands were produced from 10 primers with an average of 3.8 perRAPD primer, amplified bands were ranging from 150-950 base pair, the highest seven number of bands found in primerCLT53 (50-875 bp and lowest one band in CLT 112 (575 bp and CLT 192 (650 bp primers. All the primers were found to bemonomorphic and no genetic variation was detected within the micropropagated plants. Present protocol may be used fortesting of ginger propagules to ensure mass production of disease free planting materials.
Full Text Available 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 diversity studies, whereas, OPG-14 have highest Discrimination index (0.911 and may be used for identification of different accessions. The clustering analysis resulted in the formation of one group, only consisting of eight accessions while Nagour collections remained out of cluster. The diversity pattern did not show any correlation with the site of collection indicating that original introduction consisted of small sample size its spread was random.
With the sharp development of biomedicine, the protection and utilization of laboratory animal resources, and standardization of laboratory animal have been regarded as the theme of laboratory animal science. Many kinds of genetic markers were used in genetic background analysis, identification of species and strains, and breeding of laboratory animal, unlike morphological marker, cytological marker and isozyme marker, which revealed the difference at gene expressing level, molecular genetic markers used as an accurate and effective method reveal genetic variations of hereditary material, it plays an important role in many fields. The RAPD (Random Amplification Polymorphic DNA) method was firstly described in 1990, it has several distinct advantages over other techniques in that it is easy to perform and fast, and so was widely applied to taxonomy and identification of species, construction of genetic mapping, genetic relationships among species and population genetics. This paper summarized the application of RAPD markers in laboratory animal in recent 10 years.
García, C. B.
Full Text Available The red-legged partridge (Alectoris rufa is a typical avian species of the Mediterranean area. Crossings of red-legged partridges with other Alectoris partridges have been done in order to increase the productivity of this species when captive bred. Restocking of hunting grounds with such hybrid partridges could damage the genetic heritage of the pure red-legged partridge in the wild. In order to avoid this problem RAPD-PCR has been developed to identify hybrid individuals. This methodology has been used to study wild red-legged partridges as well as partridges from different farms. RAPD analysis results in different patterns for different species. Hybrids can be detected among presumed red-legged partridges if they present a diagnostic band which characterises another species.
Full Text Available Due to the environmental changes and habitat destruction the mudskipper fish population is decreasing in recent years. To predict the fish population structure, frequent manual survey and molecular methods are widely used. Molecular markers such as RAPD, microsatellite, allozyme, D-loop haplotype are frequently adopted to assess the population structure of an organism. In this study ten- arbitrary primers were screened to estimate the genetic relationships and diversity of two mudskipper species (Boleophthalmus boddarti and B. dussumieri in Vellar estuary, Tamilnadu, India. By this RAPD marker study, the genetic diversity (H in B. boddarti was more (0.0116 ± 0.0066 than in B. dussumieri (0.0056 ± 0.0024 in Vellar estuary (India. The genetic distance between B. boddarti and B. dussumieri was 1.7943. By observing the species specific bands and the phylogenetic analysis it is revealed that these two species clearly deviated into separate clusters emphasizing the distinct species status.
Chelomina, G N; Rozhkovan, K V; Ivanov, S A
RAPD-PCR analysis of 46 individuals of sturgeons from Amur River has been carried out. Genetic status of Amur sturgeon Acipenser schrenckii Brandt, 1869 and kaluga Huso dauricus Georgi, 1775 native populations has been estimated. Genetic evidences of hybrid origin for two phenotypical hybrids were obtained; estimations of genetic distances between species and hybrids appeared to be at interspecific level. The exact test for differentiation of populations (Exact test) and multidimensional scaling (MDS) analysis were estimated to be the most effective for species and hybrid discrimination, respectively. According to data obtained populations of sturgeon fishes which inhabit Amur River maintained an essential level of genetic variability; the presence of hybrids is regarded as one of risk factors. Multilocus RAPD-PCR markers admit as the convenient and reliable tool for genetic monitoring of Amur River sturgeons to preserve their gene pool. PMID:19140442
Eribe, Emenike Ribs K.; Olsen, Ingar
In a polyphasic approach to improve the taxonomy of Leptotrichia we have recently demonstrated heterogeneity in the enzymatic/biochemical reactions and cellular fatty acid content of Leptotrichia isolates. In the present study, SDS-PAGE of whole-cell proteins and random amplified polymorphic DNA (RAPD) analyses were used to further characterize and distinguish these isolates of which 57 had been assigned as Leptotrichia buccalis and two as ‘Leptotrichia pseudobuccalis’. The phylog...
Dutta, Suhrid R.; Kar, Prasanta K.; Ashok K. Srivastava; Sinha, Manoj K.; Jai Shankar; Ghosh, Ananta K.
The tropical tasar silkworm, Antheraea mylitta, is a semi-domesticated vanya silk-producing insect of high economic importance. To date, no molecular marker associated with cocoon and shell weights has been identified in this species. In this report, we identified a randomly amplified polymorphic DNA (RAPD) marker and examined its inheritance, and also developed a stable diagnostic sequence-characterized amplified region (SCAR) marker. Silkworms were divided into groups with high (HCSW) and l...
Mahmodi, Farshid; Kadir, J. B.; Puteh, A.; Pourdad, S. S.; Nasehi, A.; Soleimani, N.
Genetic diversity and differentiation of 50 Colletotrichum spp. isolates from legume crops studied through multigene loci, RAPD and ISSR analysis. DNA sequence comparisons by six genes (ITS, ACT, Tub2, CHS-1, GAPDH, and HIS3) verified species identity of C. truncatum, C. dematium and C. gloeosporiodes and identity C. capsici as a synonym of C. truncatum. Based on the matrix distance analysis of multigene sequences, the Colletotrichum species showed diverse degrees of intera and interspecific ...
Molecular techniques have become reliable and are highly suitable tools for identifying pathogen species and for genetic variation. The molecular marker is a useful tool for assessing genetic variations. RAPD (Random Amplified Polymorphic DNA) markers have been used to characterize the numerous filamentous fungi collected from different fields of experimental mycology. Rhizoctonia solani is a plant pathogenic fungus which cause sheath blight in rice. Present work focused on polymorphic identi...
Vellaichamy RAMANADEVI; Muthusamy THANGARAJ; Anbazhagan SURESHKUMAR; Jayachandran SUBBURAJ
Due to the environmental changes and habitat destruction the mudskipper fish population is decreasing in recent years. To predict the fish population structure, frequent manual survey and molecular methods are widely used. Molecular markers such as RAPD, microsatellite, allozyme, D-loop haplotype are frequently adopted to assess the population structure of an organism. In this study ten- arbitrary primers were screened to estimate the genetic relationships and diversity of two mudskipper spec...
Maria Sueli Papa Leuzzi; Fernanda Simões de Almeida; Mário Luís Orsi; Leda Maria Koelblinger Sodré
In this study, the RAPD technique was used to analyze the genetic structure of populations of the fish Astyanax altiparanae (Characidae, Tetragonopterinae) living in the lower, middle and upper Paranapanema River, Brazil. The aim was to assess this structure regarding fish handling and conservation programs. The genetic variability (P) was found to be 42.64%, 75% and 75% in the low, middle and upper reaches, respectively. The dendrogram of genetic similarity, obtained by comparative analysis ...
Cassava is an important tuber crop grown in the tropical and subtropical regions. Recently, we developed protocols for efficient somatic embryogenesis using zygotic embryos and nodal axillary meristems in order to reduce the genotype effect. Thereafter flow cytophotometry and randomly amplified polymorphic DNA (RAPD) markers were used to assess the ploidy level and the genetic fidelity of cassava plants regenerated by somatic embryogenesis. No change in the ploidy level of the regenerated plants was observed in comparison with the control plants. In the same way, monomorphic profiles of RAPD were obtained for the different cassava plants regenerated by somatic embryogenesis. The genetic analysis of calli showed only a few differences. Using two pairs of heterologous micro satellite primers developed in a wild African grass, a monomorphic pattern was also detected. Moreover, cultivars of different origins were also analysed using these PCR techniques. Our data from RAPD and materialistic analyses suggested that these techniques can be efficiently used to detect genetic variations in cassava. (author)
KUSUMADEWI SRI YULITA
Full Text Available Nepenthes spp. (Nepenthaceae is one of the most popular ornamental plants in Southeast Asia. There are 97 species of Nepenthes to which 64 are found in Indonesia with the center of its diversity located in Borneo. N. x hookeriana was hypothesised to be a natural hybrid between N. ampullaria and N. rafflesiana on the basis of morphological characters. Several variants of each species were also known. This present study aimed to detect the occurrence of hybrid within N. x hookeriana ‘spotted’ and ‘green’ variant using random amplified polymorphic DNA (RAPD and inter-simple sequence repeats (ISSR. Five RAPD primers and three ISSR primers were used to amplify total DNA genome and produced 83 polymorphic bands ranging in size from 300-1700 bp. Clustering analysis was performed based on RAPD and ISSR profiles using the UPGMA method. The genetic similarity of the combined markers range between 0.30-0.75 indicating a narrow range of genetic similarity among the accessions. Results from cluster analyses suggested that N. x hookeriana was indeed a hybrid between N. ampullaria and N. Rafflesiana, however it was genetically more similar to N. raflessiana.
Full Text Available Molecular techniques have become reliable and are highly suitable tools for identifying pathogen species and for genetic variation. The molecular marker is a useful tool for assessing genetic variations. RAPD (Random Amplified Polymorphic DNA markers have been used to characterize the numerous filamentous fungi collected from different fields of experimental mycology. Rhizoctonia solani is a plant pathogenic fungus which cause sheath blight in rice. Present work focused on polymorphic identification and characterization of Rhizoctonia solani isolate. Twenty eight samples were collected from different locations of South India and Punjab. Rhizoctonia solani (teleomorph: Thanatephorus cucumeris were isolated and used polymorphic examination by molecular markers. Molecular analysis was done with OPC-5, OPC-2, OPA-8 and OPA-11primers and the variability of isolated fungus DNA, allowed the visualization of 265 polymorphic bonds with molecular weight ranging from 0.5kb to 20kb significant differences in RAPD profiles of 28 isolates of R. Solani were found with two primers OPC-5 and OPC-2. To analyze the resolving ability of these primers, cumulative RAPD profiles generated by the primers were analyzed by UPGMA. The dendrogram constructed using 265 polymorphic bonds obtained from 28 isolates with 5 primers was divided into 7 clusters. Based on these results it was concluded that there was a molecular variability among the isolates of R. solani was depicted.
Mahmodi, Farshid; Kadir, J B; Puteh, A; Pourdad, S S; Nasehi, A; Soleimani, N
Genetic diversity and differentiation of 50 Colletotrichum spp. isolates from legume crops studied through multigene loci, RAPD and ISSR analysis. DNA sequence comparisons by six genes (ITS, ACT, Tub2, CHS-1, GAPDH, and HIS3) verified species identity of C. truncatum, C. dematium and C. gloeosporiodes and identity C. capsici as a synonym of C. truncatum. Based on the matrix distance analysis of multigene sequences, the Colletotrichum species showed diverse degrees of intera and interspecific divergence (0.0 to 1.4%) and (15.5-19.9), respectively. A multilocus molecular phylogenetic analysis clustered Colletotrichum spp. isolates into 3 well-defined clades, representing three distinct species; C. truncatum, C. dematium and C. gloeosporioides. The ISSR and RAPD and cluster analysis exhibited a high degree of variability among different isolates and permitted the grouping of isolates of Colletotrichum spp. into three distinct clusters. Distinct populations of Colletotrichum spp. isolates were genetically in accordance with host specificity and inconsistent with geographical origins. The large population of C. truncatum showed greater amounts of genetic diversity than smaller populations of C. dematium and C. gloeosporioides species. Results of ISSR and RAPD markers were congruent, but the effective maker ratio and the number of private alleles were greater in ISSR markers. PMID:25288981
Full Text Available The Random Amplified Polymorphic DNA (RAPD technique was used to study the genetic diversity of four Elops machnata populations in South India. Elops machnata is considered as a least concern species (LC, categorized by the International Union for Conservation and Nature (IUCN. The population trends are currently stable in Indian Ocean, Eastern Africa, but are unknown throughout the rest of its expansive range, especially in Indian estuaries. Among the ten RAPD primers tested, eight primers got amplified and gave scorable bands. In total, 119 scorable bands were observed in all populations. The overall observed and effective number of alleles was found to be 2.000 ± 0.000 and 1.5307 ± 0.2503 respectively for the entire population. The overall polymorphic loci were 61.00% and the overall gene flow among the four populations was predicted to 0.1032. The genetic distance and geographic distance between the four populations showed a positive correlation. The highest genetic similarity (0.6824 was found between Parangipettai and Muthupettai population, which reflected the geographical relationship between them. Tow main clusters were obtained based on UPGMA dendrogram. This study proves that RAPD analysis has the ability to discriminate E. machnata populations in South Indian coastal waters.
Full Text Available Genetic diversity of two wild Kalibaus, Labeo calbasu populations and one hatchery stock was studied using random amplified polymorphic DNA (RAPD method. The three 10–mer random primers (OPA01, OPB02 and OPC03 yielded a total of 26 reproducible and consistently scorable RAPD bands of which 15 (57.69% were considered as polymorphic (P95 indicating a high level of genetic variation in all the studied populations. Among the three populations, Padma population shows low level of genetic diversity (0.1238 compared to other two and it might be caused by habitat degradation in many ways which ultimately affects the genetic variation of Kalibaus. The UPGMA dendrogram based on Nei’s (1972 original measures of genetic distance (D indicated the segregation of two wild and hatchery populations of L. calbasu into two distinct clusters: the Hatchery and Padma populations produced one cluster whereas the Jamuna population belonged to another cluster. This indicates that hatchery brood stock is derived from Padma River. Nevertheless, the preliminary study revealed that RAPD technique could be an effective tool in the assessment of population genetic structure of Kalibaus.
H. S. Ginwal; Neha Mittal; Arvind Tomar; V. K. Varshney
Acorus calamus is a highly valued medicinal plant with globaldistribution used in several drugs of health care systems. We evaluatedthe genetic diversity and population structure of 50 populations of A.calamus from different geographical regions in India through RAPD andchloroplast microsatellite markers. From the total screened 82 RAPDprimers and 18 cpSSR primers, 10 RAPD and nine cpSSRs were foundpolymorphic. The selected 10 RAPD primers produced a total of 96reproducible bands, out of which 65 were polymorphic (67.70%).Whereas, the selected nine cpSSR markers produced 26 alleles and all ofthem were polymorphic. The mean genetic diversity (H) among popula-tions using RAPD (H= 0.263) and cpSSR (H=0.530) markers washigher in comparison to the mean genetic diversity within populations.Mean coefficient of gene differentiation (G) between the populationswas also high for both RAPD (G=0.830) and cpSSR markers (G=0.735), whereas the estimated gene flow was very low for RAPD (Nm =0.102) and for cpSSR (Nm = 0.179). AMOVA analysis revealed thatmore genetic variation resided among the populations than within popu-lations. Significant differences (p < 0.001) were observed between thepopulations and individuals within the populations. Cluster analysis ofRAPD and cpSSR data using UPGMA algorithm based on Nei's geneticsimilarity matrix placed the 50 populations into two main clusters. Theimplication of the results of this study in devising strategy for conserva-tion of A. Calamus is discussed.
Maria do Desterro M dos Santos
Full Text Available Onion is a crop of significant socioeconomic importance to Brazil. Onion germplasm with adaptation to tropical and sub-tropical conditions has played an important role in the development of this crop in the country. In this context, we studied the genetic diversity in a germplasm collection potentially useful for the development of cultivars for tropical and subtropical regions. The genetic variability of 21 accessions/cultivars that have been used as germplasm and/or were developed by onion breeding programs in Brazil was evaluated via RAPD markers. The following accessions were included in the study :'Red Creole', 'Roxa IPA-3', 'Valenciana 14', 'Beta Cristal', 'Diamante', 'Composto IPA-6', 'Aurora', 'Bojuda Rio Grande', 'Alfa Tropical', 'Pêra IPA-4', 'Primavera', 'Belém IPA-9', 'Crioula Alto Vale', 'Conquista', 'Pira-Ouro', 'Vale-Ouro IPA-11', 'Franciscana IPA-10', 'Serrana', 'CNPH 6400', 'Petroline', and 'Baia Periforme'. From the 520 primers used in the initial screening only 38 displayed stable polymorphisms. They produced 624 amplicons, of which 522 (83.7% were monomorphic and 102 (16.3% were polymorphic. An average similarity coefficient of 0.72 was calculated among accessions based upon this subgroup of polymorphic amplicons. This allowed the discrimination of this germplasm collection into six groups with only one of them comprising more than one accession. The main group was formed by 16 accessions ('Diamante', 'Composto IPA-6', 'Aurora', 'Bojuda Rio Grande', 'Conquista', 'Pira-Ouro', 'Serrana', 'Vale-Ouro IPA-11', 'Baia Periforme', 'Primavera', 'Franciscana IPA-10', 'Belém IPA-9', 'Crioula Alto Vale', 'Petroline', 'Pêra IPA-4' and 'Alfa Tropical', for which the genetic origin (with few exceptions can be traced back to the variety 'Baia Periforme'. The populations 'Red Creole', 'Roxa IPA-3', 'Beta Cristal', 'CNPH 6400', and 'Valenciana 14' comprised a set of five isolated groups, showing genetic divergence among them and in
In previous studies, it was found that certain RAPD (Randomly Amplified Polymorphic DNA) markers were present at higher frequencies in radionuclide-contaminated mosquitofish (Gambusia affinis) populations than in reference populations. These markers will be referred to as contaminant specific markers. In the present study, fish with and without these markers were collected from non-contaminated populations and exposed in situ to radionuclides by caging them in one of the contaminated sites. Forty fish were exposed for 1--6 weeks, after which the survivors were collected and DNA was extracted for genotypic analysis. In one experiment, the frequencies of contaminant specific markers in the survivors were compared to the frequencies of these markers in the native contaminated and uncontaminated (the source of the caged fish) populations. It was found that the genotypic distributions were more similar to the native contaminated population. In another experiment, samples of caudal fin tissue were collected for DNA extraction before and after placing fish in the cages, in order to compare survival rates of different genotypes. It was found that fish with the contaminant indicative bands had higher percent survival than the other fish. Experiments are underway or are being planned in order to determine the molecular identity of these bands and the ecological significance of altered band frequencies in hopes of developing population-level biomarkers of contaminant exposure and ecological affects
Full Text Available Lactobacillus plantarum 2035 and Lactobacillus plantarum ACA-DC 2640 are two lactic acid bacteria (LAB strains that have been isolated from Feta cheese. Both display significant potential for the production of novel probiotic food products. The aim of the present study was the development of an accurate and efficient method for the molecular detection and identification of the above strains in a single reaction. A multiplex PCR assay was designed for each strain, based on specific primers derived from Random Amplified Polymorphic DNA (RAPD Sequenced Characterized Amplified Region (SCAR analysis. The specificity of the assay was tested with a total of 23 different LAB strains, for L. plantarum 2035 and L. plantarum ACA-DC 2640. The multiplex PCR assay was also successfully applied for the detection of the above cultures in yogurt samples prepared in our lab. The proposed methodology may be applied for monitoring the presence of these strains in food products, thus evaluating their probiotic character. Moreover, our strategy may be adapted for other novel LAB strains with probiotic potential, thus providing a powerful tool for molecular discrimination that could be invaluable to the food industry.
ZHENG Guo-sheng; CHEN Xiang-ming; MENG Li
Random amplified polymorphic DNA (RAPD) was used to analyze genetic polymophism of 35Tree Peony cultivars with 7 different color groups. Thirty four primers amplified 418 DNA fragments and 337polymorphic bands (80.6%), including specific DNA markers for 18 cultivars that could be used to differentiate cultivars. The UPGMA method was used to analyze the genetic relationship among cultivars. The results showed that 35 Peony cultivars could be divided into 2 cluster groups when using similarity criteria of 1.5, and into 4 cluster groups when using similarity criteria of 1.0. The result confirmed that the flower color has no relation to the genetic clusters and the Tree Peony cultivars originated from the same area has close genetic relationship. Therefore, genetic background has no large effect on the genetic relationship. The sequence based on polymorphic rate from high to low was Blue groups ＞ Yellow groups ＞ Bark red groups ＞ Blake groups ＞White groups ＞ Green groups ＞ Red groups.
Meena Bakshi; Arvind Sharma
We studied the genetic polymorphism among 29 clones of shisham (Dalbergia sissoo Roxb) belonging to different geographic regions using random amplified polymorphic DNA (RAPD) markers. Out of 30 primers used, only 20 primers generated polymorphism in amplified product. In total 232 bands were amplified with 20 primers, of which 192 (82%) were polymorphic with an average of 9.6 bands/primer. The resolving power (Rp) ranged from 2.14 (Primer 5) to 11.93 (Primer 4). Primer 4 and Primer 3 possessed high Rp value. Polymorphism in- formation content (PIC) ranged from 0.15 (Primer 5) to 0.37 (Primer 4). Primer 4 amplified total 18 bands in 29 genotypes with PIC value of 0.37 hence; this set of primer was most informative. The similarity co- efficient analysis revealed two clusters. The first cluster comprised of only 10 clones and the second major cluster comprised of 19 clones. The genetic similarity among 29 clones ranged from 25.86% (clone 10 and 235) to 100% (clone 19 and 59), suggesting a wide genetic base in shisham clones.
M. A. Rahman
Full Text Available This experiment was conducted in the Genetics, Breeding and Reproductive Biotechnology Laboratory under Goat and Sheep Production Research Division, Bangladesh Livestock Research Institute (BLRI, Saver, Dhaka, Bangladesh. DNA was extracted from 14 goat breeds and the extracted DNA was observed by gel electrophoresis. Eight goat specific primers were synthesized by ASM-800 DNA synthesizer and screened in the study and all these primers were capable of priming polymorphic amplification pattern in both the breeds. Random amplification of polymorphic DNA â Polymerase Chain Reaction (RAPD-PCR analysis was carried out using DNA samples of 14 black bengal goat and Jamuna pari goat breeds. Only unambiguous, reproducible and scorable polymorphic fragments were taken into consideration for analysis. Data were analyzed by using a computer programe POPGENE (Version 1.31. Highest level of Nei's gene diversity value (0.4898 was observed in BMS 1494 locus and the mean genetic diversity was obtained 0.3724 among the 14 goat breeds. The highest number of polymorphism obserbed in primer BM1818.The pair-wise genetic distance value ranged from 0.2500 to 1.000. Dendrogram based on Nei's genetic distance using Unweighted Pair Group Method of Arithmetic Means (UPGMA indicated segregation of the 14 goat breeds. Within Jamuna pari goat genetic similarity is low as well as black Bengal goat.
The contents of phycocyanin (PC), allo-phycocyanin (APC), phycobiliproteins (PBP) and the ratio of PC and APC (PC/APC) of nine Spirulina platensis strains used in pilot plant cultivation were 10.74%-16.26%, 3.67%-5.55%, 14.42%-21.81% and 2.87-3.05, respectively. By mechanical grinding and sedimentation, the single cells or spheroplasts of Sp-CH, whose PBP contents was highest in the nine tested strains, were obtained. They were treated by 0.6% EMS for 30 min and 2.4 kGy 60Co γ-rays, and then cultured by the single filament separating, a high PBP content strain named as Sp-CH32 was obtained. The contents of PC, APC and PBP of Sp-CH32 were about 36%, 89% and 50%, higher than that of Sp-CH, respectively. And the PC/APC of Sp-CH32 were 1.91-2.23. Moreover, RAPD analysis showed that the amplifcation bands of primers S38 revealed significant polymorphisms in Sp-CH32 and Sp-CH. The pilot plant cultivation in 3 consecutive years showed that Sp-CH32 was a perfect high PBP content strain with excellent characteristics, and has been applied in mass cultivation and industrialization. (authors)
SHANG Huimin; SONG Weibo
On the basis of the random amplified polymorphic DNA-fingerprinting (RAPD) method, 10 morphospecies of scuticociliates from 7genera, including 15 clones of 13 strains, Uronema rnarinum, Uronema cf marinum, Parauronerna virginianum, P. longurn, Metanophrys similis, M. sinensis, Paralembus digitiformis, Mesanophrys carcini, Paranophrys magna and Cohnilembus verminus were analysed using 8 oligonucleotide primers. The genetic similarity among the clones of the same strain measured by a band-sharing index is 0.97～0.98,while 0.40～0.52 among strains. This value measured is 0.39～0.46 among congeners of the same genus, whereas 0.16～0.47 between different genera. A distance tree was constructed based on 8-primer analysis, in which the scuticociliates investigated were separated into two clusters: one consists of 2 genera, Uronema and Parauronema, and the other was composed of five, Metanophrys, Mesanophrys,Paranophrys, Paralembus and Cohnilembus. It is demonstrated also that the morphospecies Parauronema virginianum may be a species-complex, i. e., it contains different genospecies.
Full Text Available Rubber hand pollination in Indonesian Rubber Research Institute program currently uses Wickham population which genetic analysis showed that genetic diversity of this population is narrow. The development of breeding activity has made the genetic base narrower by inbreeding. In order to solve this problem can use a new genetic resource that is the rubber germplasm IRRDB 1981 population. The genetic relationship between these populations is important to choose parents to avoid closely related genotypes in hand pollination. Therefore RAPD analysis was carried out using four selected primers i.e. OPH-03, OPH-05, OPH-18 and OPN-06. The result showed that Wickham and IRRDB 1981 population were separated into two different big groups with genetic similarity value of 0.64, and those big groups were separated further into many small sub groups with some genetic similarity level. The genetic similarity matrix showed that Wickham and IRRDB 1981 population has a range of genetic similarity 0.37– 0.98. The highest genetic similarity was found between RRIM 600 and PN 621, while the lowest was between BPM 1 and RRIC 100. Value in this matrix showed the genetic diversity between each clone. Based on this result, rubber genotypes of Wickham population could be crossed with genotypes of IRRDB 1981 population by choosing genotypes that have low genetic similarity.
Organogenesis was induced in callus derived from mature zygotic embryos of six families (J-56, S-1003, E-22, E-311, E-440, and Mc) of loblolly pine (Pinus taeda L.) within 24 weeks of culture. Elongation of adventitious buds was achieved on TE medium supplemented with 0.5 mg·L-1 indole-3-butyric acid (IBA) and 1 mg/l 6-benzyladenine (BA). The most suitable medium for root formation proved to be TE medium supplemented with 0.5 mg·L-1 IBA, 2 mg·L-1 BA , and 0.5 mg/l gibberellic acid (GA3). 169 regenerated plantlets were transferred to a perlite∶peatmoss∶vermiculite (1:1:1) soil mixture, and 98 plantlets survived in the field. Total DNA was extracted from the needles of the regenerated plantlets of the six families of loblolly pine. Analysis of random amplified polymorphic DNA (RAPD) using 20 arbitrary oligonucleotide 10-mers, show that amplification products were monomorphic for all the plantlets of family J-56, S-1003, E-22, E-311, E-440, and Mc of loblolly pine. These results suggested that organogenesis can be used for clonal micropropagation of some families of loblolly pine.
Variability obtained from mutation breeding (gamma rays) was examined through molecular marker techniques (RAPD). A total of 85 loci were amplified, out of which 76.47% were polymorphic and 23.53% were monomorphic. Fragments size ranged from 220bp-2.1kb and fragments produced by various primers ranged from 3-13 with an average of 5 fragments per primer. The highest number of loci (13) was amplified with primer B-07, while the lowest number 3 with primer B-01. Results revealed that mutant P1 (20Gy) contained a specific segment of 2.03kb. Genetically most similar genotypes were P2 (10Gy) and P4 (20Gy) (95.55%) while most dissimilar genotypes were P4 (10Gy) and P3 (20Gy) (63.2%). On the basis of results achieved, the mutants could be divided into four clusters and three groups. Mutants P4 (40 Gy) and P4 (10 Gy) were genetically distinct from other mutants. (author)
Bekessy, Sarah A; Allnutt, T R; Premoli, A C; Lara, A; Ennos, R A; Burgman, M A; Cortes, M; Newton, A C
Araucaria araucana (Monkey Puzzle), a southern South American tree species of exceptional cultural and economic importance, is of conservation concern owing to extensive historical clearance and current human pressures. Random amplified polymorphic DNA (RAPD) markers were used to characterise genetic heterogeneity within and among 13 populations of this species from throughout its natural range. Extensive genetic variability was detected and partitioned by analysis of molecular variance, with the majority of variation existing within populations (87.2%), but significant differentiation was recorded among populations (12.8%). Estimates of Shannon's genetic diversity and percent polymorphism were relatively high for all populations and provide no evidence for a major reduction in genetic diversity from historical events, such as glaciation. All pairwise genetic distance values derived from analysis of molecular variance (Phi(ST)) were significant when individual pairs of populations were compared. Although populations are geographically divided into Chilean Coastal, Chilean Andes and Argentinean regions, this grouping explained only 1.77% of the total variation. Within Andean groups there was evidence of a trend of genetic distance with increasing latitude, and clustering of populations across the Andes, suggesting postglacial migration routes from multiple refugia. Implications of these results for the conservation and use of the genetic resource of this species are discussed. PMID:11920130
Galanis, Alex; Kourkoutas, Yiannis; Tassou, Chrysoula C.; Chorianopoulos, Nikos
Lactobacillus plantarum 2035 and Lactobacillus plantarum ACA-DC 2640 are two lactic acid bacteria (LAB) strains that have been isolated from Feta cheese. Both display significant potential for the production of novel probiotic food products. The aim of the present study was the development of an accurate and efficient method for the molecular detection and identification of the above strains in a single reaction. A multiplex PCR assay was designed for each strain, based on specific primers derived from Random Amplified Polymorphic DNA (RAPD) Sequenced Characterized Amplified Region (SCAR) analysis. The specificity of the assay was tested with a total of 23 different LAB strains, for L. plantarum 2035 and L. plantarum ACA-DC 2640. The multiplex PCR assay was also successfully applied for the detection of the above cultures in yogurt samples prepared in our lab. The proposed methodology may be applied for monitoring the presence of these strains in food products, thus evaluating their probiotic character. Moreover, our strategy may be adapted for other novel LAB strains with probiotic potential, thus providing a powerful tool for molecular discrimination that could be invaluable to the food industry. PMID:26506345
The present study was amid to investigate the genetic diversity of twenty peach cultivars and genotypes by RAPD primers at the Institute of Biotechnology and Genetic Engineering, KPK Agricultural University Peshawar. The result indicated that fifteen primers (GLCO9, GLC20, GLA20, GLA13, GLB10, GLB20, GLB06, GLB19, GLA19, GLB19, GLD16, GLB15, GLA15, GLB12, GLB11) gave genetic distance among the peach cultivars and genotypes under study by PCR amplification. Average genetic diversity (estimated as genetic distance) ranged between 12 and 58%. The molecular size of most of the bands were from 150 bp to 1000 bp. Based on dendrogram analysis, Khyber 1 and Khyber 2 was grouped in cluster A, and Tex-A6-69 and BY-8-135 in cluster B, Candan and 6A were most closely related cultivars and genotypes among the 20 peach cultivars and genotypes while Lering, Flam crest, Tex x-9, early grand and Floradaking were distinctly grouped when compared with the rest of population. (author)
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
Asma Bettaibi; Maha Mezghani-Khemakhem; Dhia Bouktila; Hanem Makni; Mohamed Makni
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 polym...
Kenia Gracielle da Fonseca
Full Text Available O Brasil é o maior produtor mundial de maracujá, entretanto tem-se observado redução na produtividade do maracujazeiro nos últimos anos, devido, principalmente, a fatores fitossanitários. Na Embrapa Cerrados, a transferência de genes de resistência de espécies silvestres para as comerciais de maracujazeiro tem sido feita por meio de hibridações interespecíficas seguidas de um programa de retrocruzamentos auxiliados por marcadores moleculares. Este trabalho teve por objetivo verificar a recuperação do genoma recorrente nas plantas RC4 e RC5 [(Passiflora edulis x Passiflora setacea x Passiflora edulis ] com base em marcadores RAPD. O estudo foi desenvolvido no Laboratório de Genética e Biologia Molecular da Embrapa Cerrados. Amostras de DNA de cada material genético (17 plantas RC4, 16 plantas RC5, Passiflora edulis e Passiflora setacea foram amplificadas para obtenção de marcadores RAPD. Foram utilizados 12 primers decâmeros para as plantas RC4 e 14 primers decâmeros para as plantas RC5. Os marcadores RAPD gerados foram convertidos em matriz de dados binários. Verificou-se alta porcentagem de marcadores polimórficos em consequência do cruzamento-base interespecífico. A menor similaridade genética foi observada entre as espécies P. edulis e P. setacea, evidenciando a grande distância genética dessas espécies.Brazil is the largest world producer of passion fruit, however, it has been observed a reduction in the productivity in recent years due, mainly, to phytosanitary factors. At Embrapa Cerrados, the transfer of resistance genes from wild to commercial species of passion fruit has been made through interspecific hybridations, followed by a backcrossing molecular marker-assisted program. The objective this work was to verify the recovery of recurrent genome at the plants RC4 and RC5 [(Passiflora edulis x Passiflora setacea x Passiflora edulis] based on RAPD markers. The study was developed at Embrapa Cerrados
Badirzadeh, Alireza; Mohebali, Mehdi; 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 identified by nested-PCR assay. The disease was confirmed in 20 out of 30 (66%) suspected patients by parasitological examinations. L. infantum was identified in eight and Leishmania major in 12 CL cases by nested-PCR. Cutaneous leishmaniasis patients infected with L. major had the history of travel to CL endemic areas. L. infantum antibodies were detected by direct agglutination test (DAT) at titers of 1:3200 in two cases with history of VL. Results of this study indicated that L. infantum is a causative agent of CL as well as PKDL in the VL endemic areas. PMID:23816511
Verma, Sandeep; Avishek, Kumar; Sharma, Vanila; Negi, Narendra Singh; Ramesh, Venkatesh; Salotra, Poonam
Loop-mediated isothermal amplification (LAMP) is at the forefront in the search for innovative diagnostics for rapid and specific amplification of target DNA under isothermal conditions. We have applied LAMP assay using SYBR Green for clear-cut naked eye detection of Leishmania (Leishmania) donovani in 200 clinical samples of visceral leishmaniasis (VL) and post-kala-azar dermal leishmaniasis (PKDL). The assay was positive in 53/55 VL blood samples (sensitivity, 96.4%; 95% confidence interval [CI], 87.7-99%), 15/15 VL bone marrow aspirate samples (sensitivity, 100%; 95% CI, 79.6-100%), 60/62 PKDL tissue biopsy samples (sensitivity, 96.8%; 95% CI, 88.9-99.1%), and 1/68 control samples (specificity, 98.5%; 95% CI, 92.1-99.7%). The assay was specific for L. (L.) donovani, the causative species for VL and negative for L. (L.) infantum, L. (L.) tropica, and L. (L.) major. This is the first comprehensive clinical study demonstrating the applicability of the LAMP assay for a rapid and reliable molecular diagnosis of VL and PKDL. PMID:23433714
Full Text Available In the next paper I intend to look the novel written by Manuel Vicent El azar de la mujer rubia. The text is presented to us as a new perspective on the transition, this time structured from a female character from which are drawn the wires of the Spanish history since Franco’s death until the present. However, the reading of the work reveals that the character is in the foreground is not another that Adolfo Suárez and that the material that serves as a starting point is the own journalistic of Manuel Vicent. It would think then this novel as an attempt to save from oblivion those newspaper articles of inestimable value –for having been written at the time that the historical events were unfolding– but also as a work that was presented as novel to reveal the hidden story of a love triangle that directly influenced the history of the country, with a female character that is, for the first time, recovered and processed into heroin. The study will focus on the way in which Vicent retrieves those journalistic texts and in the operations of inclusion of the same in a text that mixes fiction with history to return, once more, to the one of the momentous of the recent Spanish history
Mohammad Shafiul Alam
Full Text Available Mymensingh is the most endemic district for kala-azar in Bangladesh. Phlebotomus argentipes remains the only known vector although a number of sand fly species are prevalent in this area. Genotyping of sand flies distributed in a VL endemic area was developed by a PCR and restriction-fragment-length polymorphism (RFLP of 18S rRNA gene of sand fly species. Using the RFLP-PCR analysis with AfaI and HinfI restriction enzymes, P. argentipes, P. papatasi, and Sergentomyia species could be identified. Among 1,055 female sand flies successfully analyzed for the species identification individually, 64.4% flies was classified as Sergentomyia species, whereas 35.6% was identified as P. argentipes and no P. papatasi was found. Although infection of Leishmania within the sand flies was individually examined targeting leishmanial minicircle DNA, none of the 1,055 sand flies examined were positive for Leishmania infection. The RFLP-PCR could be useful tools for taxonomic identification and Leishmania infection monitoring in endemic areas of Bangladesh.
Newton Alex Mayer
Full Text Available Um projeto de pesquisa visando à utilização de clones de umezeiro (Prunus mume Sieb. et Zucc. como porta-enxertos para pessegueiro [Prunus persica (L. Batsch] está sendo conduzido na FCAV/UNESP, Câmpus de Jaboticabal-SP, com promissoras perspectivas de sucesso. Três genótipos de umezeiro foram selecionados de acordo com características agronômicas desejáveis para esta finalidade. A distinção dos três genótipos entre si, baseada exclusivamente em características morfológicas, apresenta limitações. Dessa forma, o objetivo do presente trabalho foi identificar marcadores RAPD capazes de diferenciar e caracterizar os Clones 05, 15 e a cv. Rigitano (Clone 10 de umezeiro, utilizando-se das cultivares Aurora-1 e Okinawa de pessegueiro como outgroup. Dos 220 primers testados, foram selecionados 42, que amplificaram todos os cinco genótipos. Verificou-se que os marcadores RAPD permitiram a distinção entre o Clone 05, o Clone 15 e a cv. Rigitano de umezeiro, demonstrando a existência de variabilidade genética entre os mesmos. Dentre os três genótipos de umezeiro estudados, constatou-se que a similaridade genética é maior entre o Clone 05 e o Clone 15.A research project with the objective do develop mume clones (Prunus mume Sieb. et Zucc., to be used as rootstocks for peach tree [Prunus persica (L. Batsch] is been carried out at the Faculdade de Ciências Agrárias e Veterinárias (FCAV/UNESP, Jaboticabal Campus, São Paulo State, Brazil. These project showed promising perspectives of success, with three clones that were selected according to their characteristics for peach rootstock. But the distinction of the three clones among them, based only in morphologic characteristics, has presented limitations. The objective of the present research was to identify RAPD markers able to characterize and differentiate the 05 and 15 Clones and Rigitano mume cultivar, using Aurora-1 and Okinawa peach tree as outgroup. Among the 220 tested
Ana Paula de Andrade Aukar
Full Text Available It has been evaluated the genetic variability through the use of RAPD molecular markers on the following passionflower species: Passiflora amethystina, P. caerulea, P. cincinnata, P. coccinea, P. serrato digitata, P. foetida, P. maliformis, P. alata, P. giberti, P. laurifolia, P. macrocarpa, P. nitida, P. setacea, P. suberosa, P. ligularis, P. capsularis, P. edulis Sims and its botanical variety P. edulis Sims f. flavicarpa Deg. In this research work, the analyses of the random amplified polymorphic DNA products (RAPD were employed to estimate the genetic diversity and the taxonomic linkage within the species above. The total of 21 primers were used in this study which generated 270 different polymorphic products. It was possible to detect that the Passiflora species had shown a similarity of 17,3%, and between Passiflora edulis Sims and Passiflora edulis Sims f. flavicarpa a similarity of 34,35% has been found. The rate of similarity within edulis specie is low, making it clear that a large variability between the yellow and the purple forms exists.Foram avaliadas as variações genéticas através de marcadores moleculares RAPD, as seguintes espécies de maracujá: Passiflora amethystina, P. caerulea, P. cincinnata, P. coccinea, P. serrato digitata, P. foetida, P. maliformis, P. alata, P. giberti, P. laurifolia, P. macrocarpa, P. nitida, P. setacea, P. suberosa, P. ligularis, P. capsularis, P. edulis Sims e sua variedade botânica P. edulis Sims f. flavicarpa Deg. Neste estudo, a análise dos produtos da amplificação ao acaso do DNA polimórfico (RAPD foi usada para estimar a diversidade genética e as relações taxonômicas entre as espécies. Foram utilizados 21 "primers", que produziram um total de 270 bandas polimórficas. Verificou-se que as espécies de Passiflora apresentaram uma média de similaridade de 17,3%, e entre Passiflora edulis Sims e Passiflora edulis Sims f. flavicarpa, de 34,35%. Pode-se perceber que o valor de
José Hortêncio Mota
Full Text Available Realizou-se este estudo com o objetivo de determinar a diversidade genética entre doze cultivares de alho, por meio de marcadores moleculares RAPD. Foram utilizados seis cultivares nobres e seis cultivares seminobres. A análise de agrupamento das similaridades genéticas foi realizada pelo método UPGMA, gerando um dendrograma utilizando o índice de Jaccard. Houve a formação de dois grupos, sendo o primeiro grupo formado pelas cultivares nobres (Roxo Pérola de Caçador, Chonan, Contestado 12, Caçador 30, Caçador 40 e Quitéria 595, ou seja, cultivares que precisam de vernalização para a formação do bulbo, e um segundo formado pelas cultivares seminobres (Gigante Curitibanos, Gigante Roxo, Gigante Roxão, Gravatá, Amarante e Cateto Roxo ou que não precisam de vernalização para a formação do bulbo. As cultivares nobres e seminobres apresentaram 57,1% e 54,2% de similaridade, respectivamente. Pelos resultados, pode-se concluir que o marcador molecular RAPD foi eficiente em separar dois grupos de cultivares de Allium sativum.The objective of this study was to determine the genetic diversity among twelve garlic cultivars, with RAPD molecular marker. Six cultivars noble and six cultivars half-noble were tested. The analysis of grouping by genetic similarities was carried out by the method of UPGMA with generated a dendrogram using the Jaccard index. Have the formation of two groups, being the first group formed by the cultivars noble (Roxo Pérola de Caçador, Chonan, Contestado 12, Caçador 30, Caçador 40 and Quitéria 595 those that need vernalization for the formation of the bulb and a second group formed by the cultivars half-noble (Gigante Curitibanos, Gigante Roxo, Gigante Roxão, Gravatá, Amarante and Cateto Roxo which do not need vernalization for formation of the bulb. The cultivars noble and half-noble presented 57,1% and 54,2% of similarity, respectively. The results allowed to conclued that RAPD molecular marker were
Utilização de microssatélites e RAPD na caracterização molecular de acessos de Paspalum urvillei Steudel Genetic diversity among accesses of Paspalum urvillei Steudel estimated by microssatelites and RAPD markers
Joaquim Taizo Sawasato
Full Text Available Este estudo foi realizado com o objetivo de verificar a diversidade genética na coleção de acessos de P. urvillei do Departamento de Plantas Forrageiras e Agrometeorologia (DPFA da Faculdade de Agronomia (UFRG visando sua utilização em futuros trabalhos de seleção. Foram avaliados 64 acessos provenientes do Rio Grande do Sul, 1 de Xanxerê, Santa Catarina, três de Curitiba, Paraná, e 1 da Argentina. A diversidade genética foi analisada por meio de marcadores do tipo RAPD e SSR. Utilizaram-se dez primers para marcadores RAPD, o que possibilitou obter 56 bandas polimórficas e 11 grupos no dendrograma com similaridade média de 0,70. Na técnica de SSR, foram utilizados sete primers e obtidas 28 bandas polimórficas, formando sete grupos no dendrograma com similaridade média de 0,66. Ambos os marcadores foram eficientes para o agrupamento de acessos coletados. O uso de maior número de primers para gerar mais bandas polimórficas foi necessário para obtenção de fingerprintsgenômicos dos indivíduos similares. Os dendrogramas gerados neste estudo dão subsídios para futuros cruzamentos de gerações parentais contrastantes ou similares no melhoramento de Paspalum urvillei.The aim of this study was to estimate the genetic diversity among accesses of P. urvillei of Departamento de Plantas Forrageiras e Agrometeorologia (DPFA of the College of Agronomy - UFRGS and to evaluate their use in selection programs. Sixty four accesses from different cities of the Southern Region of Brazil (Rio Grande do Sul, Santa Catarina and Parana States and from Argentine were analyzed by RAPD and SSR molecular markers. Ten primers of RAPD markers were used and resulted in 56 polymorphic bands and 11 groups in a dendrogram with average similarity 0.70. Seven primers were used for the SSR technique and resulted in 28 polymorphic bands and seven groups in a dendrogram with average similarity 0.66. Both markers were efficient on grouping the accesses
Sunil Kumar, K N; Maruthi, K R; Alfarhan, A H; Rajakrishnan, R; Thomas, J
Mistletoes are hemiparasitic plants growing on aerial parts of other host trees. Many of the mistletoes are reported to be medicinally important. The hemiparasitic nature of these plants makes their chemical composition dependent on the host on which it grows. They are shown to exhibit morphological dissimilarities also when growing on different hosts. Helicanthus elastica (Desr.) Danser (mango mistletoe) is one such less explored medicinal mistletoe found on almost every mango tree in India. Traditionally, the leaves of this plant are used for checking abortion and for removing stones in the kidney and urinary bladder while significant antioxidant and antimicrobial properties are also attributed to this species of mistletoe. The current study was undertaken to evaluate molecular differences in the genomic DNA of the plant while growing on five different host trees using four random markers employing random amplified polymorphic DNA (RAPD) followed by similarity matrix by Jaccard's coefficient and distance matrix by hierarchal clustering analysis. Similarity and distance matrix data employing just 4 random markers, separately and the pooled data as well, revealed significant difference in the genomic DNA of H. elastica growing on five different hosts. Pooled data of similarity from all the 4 primers cumulatively showed similarity between 0.256 and 0.311. Distance matrix ranged from of 0.256 to 0.281 on pooling the data from all the four primers. The result employing a minimum number of primers could conclude that genomic DNA of H. elastica differs depending upon the host on which it grows, hence the host must be considered while studying or utilizing this mistletoe for medicinal purposes. PMID:27081357
庄志猛; 石拓; 孔杰; 刘萍; 刘振辉; 孟宪红; 邓景耀
The random amplified polymorphic DNA analysis was used to estimate genetic diversity in one successively cultivated stock and three wild stocks of Penaeus chinensis shrimp, two of which were collected from the spawning and wintering grounds in the west coast of Korean Peninsula, and one from the feeding ground in the China coast of the Yellow Sea. A random primer kit was employed to scan the genomic DNA in 20 individuals of each index stock. A total of 110 reproducible RAPD markers were obtained, 68.2 % of which showed a sound eonformability within all the individuals detected, implying that the genetic variability in P. chinensis is relatively low. The proportions of polymorphic loci among these four stocks ranged from 20% to 33.3%, while the degrees of genetic polymorphisms varied from 0.0093 to 0.0307. The genetic variability of inter-stocks was higher than that of intra-stock. The genetic diversity in different stocks differed from each other; that is, a less genetic differentiation in the spawning and wintering stocks from the west coast of Korean Peninsula was revealed and their genetic diversities were higher than that of the spawning stock in the Bohai Sea and the China coast of the Yellow Sea. As detected, the genetic diversity in the successively cultivated stock was the lowest among these four stocks. Through genetic distance analysis between a random pair of individuals, a dendrogram of the above-mentioned four stocks was constructed by unweighted pair group method with arithmetic mean. The results based on cluster analysis well fitted with the geographical distribution of P. chinensis in the Bohai and Yellow Seas.
Full Text Available The yeast microbiota occurring on different varieties of grapes grown in cool-climate is not completely researched. Therefore, its identification is important to research. On the other hand, yeasts occurring in these fruits can be potentially used as starter cultures to obtain particularly demanded features in the production of wine. In addition, rapid methods for yeast identification allow to eliminate the contamination with pathogenic yeasts, which could cause the loss of wine production. The aim of the study was to isolate and identify the yeasts occurring on the surface of the different varieties of white and red grapes, grown in cool-climate of Poland. Also, the aim was to compare the qualitative and quantitative composition of yeasts on the tested grapes. The 84 cultures of yeasts were isolated, that were initially macroscopic and microscopic analyzed and the purity of cultures was rated on the WL medium. Identification of yeasts by PCR-RAPD was carried using the M13 primer. In the PCR-RFLP method ITS1 and ITS4 primers, as well as restriction enzymes HhaI, HinfI, HaeIII, were used. Preliminary identification of yeasts by standard methods produced results very different from the results obtained by molecular methods. Among the isolated microorganisms yeasts were dominating, but bacteria and molds were also present. Using the PCR-RAPD method most strains of yeasts were identified. Yeast microflora of different varieties of white and red grapes was very similar as the same species of yeasts were identified. Yeasts of the genus Saccharomyces were present in all varieties of grapes. The Rhodotorula mucilaginosa, Saccharomyces cerevisiae, Metschnikowia pulcherrima, Rhodotorula minuta, Pichia kluyveri, Hanseniaspora uvarum and Rhodotorula mucilaginosa were identified by PCR-RAPD. 4 of the 33 tested strains of yeasts were identified by PCR-RFLP. By PCR-RAPD only Hanseniaspora uvarum was identified. The quantity and quality of microorganisms living
Rabello, Aline R; Paulo R. Queiroz; Kenya C.C. Simões; Hiragi, Cássia O.; Luzia H.C. Lima; Maria Regina V. Oliveira; Angela Mehta
The Bemisia tabaci complex is formed by approximately 41 biotypes, two of which (B and BR) occur in Brazil. In this work we aimed at obtaining genetic markers to assess the genetic diversity of the different biotypes. In order to do that we analyzed Bemisia tabaci biotypes B, BR, Q and Cassava using molecular techniques including RAPD, PCR-RFLP and sequencing of the ITS1 rDNA region. The analyses revealed a high similarity between the individuals of the B and Q biotypes, which could be distin...
Keize Pereira Junqueira
Full Text Available A espécie de pitaya mais cultivada atualmente é Hylocereus undatus, a pitaya-vermelha-de-polpa-branca. Colômbia e México são os principais produtores mundiais e, devido à sua rusticidade, a pitaya é considerada uma alternativa potencialmente viável também para o aproveitamento de solos pedregosos, arenosos e maciços rochosos. Apesar da crescente demanda, ainda não há uma cultivar lançada no mercado que atenda às necessidades climáticas de produção e às exigências do consumidor brasileiro. O presente trabalho é parte do programa de seleção e melhoramento da pitaya CPAC PY-01 da Embrapa Cerrados. Objetivou-se realizar o estudo da variabilidade genética de 16 acessos de pitayas mantidos na coleção de germoplasma da Embrapa Cerrados, apresentando diferentes características fenotípicas relacionadas especialmente à produção, por meio de marcadores moleculares RAPD (Random Amplified Polymorphic DNA. O DNA genômico de cada acesso foi extraído, e onze iniciadores decâmeros foram utilizados para a obtenção de marcadores moleculares RAPD, que foram convertidos em matriz de dados binários, a partir da qual foram estimadas as distâncias genéticas entre os acessos e realizadas análises de agrupamento e de dispersão gráfica. Foram obtidos 111 marcadores RAPD, perfazendo uma média de 10,1 marcadores por primer, dos quais 45 (40,54% foram polimórficos. As distâncias genéticas entre os 16 acessos variaram entre 0,006 e 0,148. As maiores distâncias genéticas foram obtidas entre os acessos "52" e "61", sendo que, em 2007, o primeiro produziu mais de 25 frutos, e o segundo, nenhum. Assim, deduz-se que, nesse caso, a próvável causa da variação seja genotípica. As menores distâncias genéticas foram constatadas entre os acessos "63"e "55" e entre "19"e "59". Os dois grupos apresentaram valores de produção próximos. Os marcadores moleculares RAPD mostraram que, mesmo dentro da mesma espécie, há variabilidade gen
In 1977, approximately 250 Mosquitofish (Gambusia affines) were transplanted from a relatively uncontaminated site into a small pond on the Oak Ridge Reservation that is heavily contaminated with radionuclides. DNA polymorphisms, using the RAPD technique, were examined in order to determine if any genetic differentiation had occurred between the two populations. Also, fish from another radionuclide-contaminated population (White Oak Lake) and two unrelated non-contaminated populations were also examined. The RAPD (Randomly Amplified Polymorphic DNA) technique uses the polymerase chain reaction with a short oligonucleotide primer to produce DNA fragments of various lengths. When analyzed by gel electrophoresis, these fragments form banding patterns similar to DNA fingerprints. A total of 26 primers were used to produce DNA band patterns, many of which revealed population differences. In addition several primers revealed banding patterns which differentiated between the Crystal Springs and Pond 3513 populations. Furthermore, bands found at high frequency in Pond 3513 and White Oak Lake populations were absent or present at a lower frequency in the non-contaminated populations. For some primers, the contaminated populations showed more DNA bands per individual, and fish with more bands had fewer DNA strand breaks than the fish with fewer bands. These data will be discussed with relation to biomonitoring programs and evolution of resistance to genotoxins in natural populations
Pineda Santis, Hermes; Arboleda Chacón, Lucy; Echeverry Echavarria, Amparo; Urcuqui Inchima, Silvio; Pareja Molina, Diego; Olivera Angel, Martha; Builes Gómez, Juan
Knowledge on the genetic diversity of wild fish species is essential for conservation and appropriate management of individuals in repopulation programs. In Colombia, Brycon henni has been reported in the Magdalena and Cauca river basins, but the population and range have diminished as a consequence of anthropic activities. In this study, the Random Amplified Polymorphic DNA (RAPD) was used to estimate the actual genetic structure in this species. For the purpose, six sample sites located in the department of Antioquia (Central Chain Mountains of Colombia) were used. Thirty five primers (87.5%), out of forty used, yielded 1 466 reliable and consistent fragments; 417 were considered as unique fragments able to discriminate among the Magdalena (Humarada-1 and Humarada-2) and Cauca (Piedras, La Clara y Guaracfi) river basins samples, suggesting that each is a discrete unit. This diversity suggests that anthropic effects of over fishing, dam building, deforestation and water pollution, have contributed to the isolation of these fish groups on the high mountains. Brycon moorei and Colossoma macropomum, as an interspecific control groups, were placed out of the B. henni general group, confirming their taxonomic classification through morphologic data. The RAPD technique was useful to know the genetic diversity and to discriminate among B. henni populations from different geographic origins, as a basis for an appropriate plan of repopulation, conservation and wildlife management. PMID:19086404
LI Dong-mei; YE Qing-sheng; ZHU Gen-fa
The random amplified polymorphic DNA (RAPD) marker was assessed to detect the genetic relationships among 48hybrid Cymbidium cultivars from Japan, Korea, China, and USA, and 2 species of native Cymbidium. Twenty primers were screened from 100 random decamer primers, and a total of 258 DNA bands were amplified, 253 of which (98.1%) were polymorphic. The average number of polymorphic DNA bands amplified by each primer was 12.6. All culfivars were distinguishable when a number of primers were considered. Genetic similarities among the cultivars and species were estimated based on the amount of band sharing ranging from 0.364-0.817 with an average of 0.581. According to the data,a dendrogram of genetic relationship, which was constructed using the UPGMA method, showed that all the tested cultivars and native species were classified into five cluster groups with the similarity coefficient of 0.592. It revealed that the genetic relationships among tested accessions were to some extent related with their origin, flower colour, branch type, and genealogy. It further indicated that the RAPD technique is a useful tool for studying the genetic relationships among hybrid Cymbidium cultivars.
Lam, Kelly Y C; Chan, Gallant K L; Xin, Gui-Zhong; Xu, Hong; Ku, Chuen-Fai; Chen, Jian-Ping; Yao, Ping; Lin, Huang-Quan; Dong, Tina T X; Tsim, Karl W K
Cordyceps sinensis is an endoparasitic fungus widely used as a tonic and medicinal food in the practice of traditional Chinese medicine (TCM). In historical usage, Cordyceps specifically is referring to the species of C. sinensis. However, a number of closely related species are named themselves as Cordyceps, and they are sold commonly as C. sinensis. The substitutes and adulterants of C. sinensis are often introduced either intentionally or accidentally in the herbal market, which seriously affects the therapeutic effects or even leads to life-threatening poisoning. Here, we aim to identify Cordyceps by DNA sequencing technology. Two different DNA-based approaches were compared. The internal transcribed spacer (ITS) sequences and the random amplified polymorphic DNA (RAPD)-sequence characterized amplified region (SCAR) were developed here to authenticate different species of Cordyceps. Both approaches generally enabled discrimination of C. sinensis from others. The application of the two methods, supporting each other, increases the security of identification. For better reproducibility and faster analysis, the SCAR markers derived from the RAPD results provide a new method for quick authentication of Cordyceps. PMID:26694332
Kelly Y. C. Lam
Full Text Available Cordyceps sinensis is an endoparasitic fungus widely used as a tonic and medicinal food in the practice of traditional Chinese medicine (TCM. In historical usage, Cordyceps specifically is referring to the species of C. sinensis. However, a number of closely related species are named themselves as Cordyceps, and they are sold commonly as C. sinensis. The substitutes and adulterants of C. sinensis are often introduced either intentionally or accidentally in the herbal market, which seriously affects the therapeutic effects or even leads to life-threatening poisoning. Here, we aim to identify Cordyceps by DNA sequencing technology. Two different DNA-based approaches were compared. The internal transcribed spacer (ITS sequences and the random amplified polymorphic DNA (RAPD-sequence characterized amplified region (SCAR were developed here to authenticate different species of Cordyceps. Both approaches generally enabled discrimination of C. sinensis from others. The application of the two methods, supporting each other, increases the security of identification. For better reproducibility and faster analysis, the SCAR markers derived from the RAPD results provide a new method for quick authentication of Cordyceps.
Hizer, Suzanne E; Dhar, Arun K; Klimpel, Kurt R; Garcia, Denise K
Random amplified polymorphic DNA (RAPD) fingerprints of two shrimp populations (Litopenaeus stylirostris) were compared to find genetic marker(s) that may be associated with infectious hypodermal and hematopoietic necrosis virus (IHHNV) resistance or susceptibility. Of the 100 10-mer random primers and 100 intersimple-sequence repeat (ISSR) primers screened, five provided markers specific to the Super Shrimp population and three provided markers specific to the wild caught population. The two populations were further characterized for relative viral load (reported as cycle threshold, CT) using real-time quantitative PCR with primers specific to the IHHNV genome. The beta-actin gene was amplified to serve as a control for normalization of the IHHNV viral load. The mean viral load was significantly lower (C(T) = 34.58; equivalent to 3.3 x 10(1) copies of IHHNV genome/ng DNA) in Super Shrimp than in the wild caught population (CT = 23.49; equivalent to 4.2 x 10(4) copies/ng DNA; P < 0.001; CT values are inversely related to viral load). A preliminary prediction model was created with Classification and Regression Tree (CART) software (Salford Systems, San Diego, Calif.), where the resultant decision tree uses the presence or absence of seven RAPD markers as predictors of the relative viral load. PMID:11908650
Rachel Fatima Gagliardi; Luiz Ricardo Hanai; Georgia Pacheco; Carlos Alberto Oliveira; Leonardo Alves Carneiro; José Francisco Montenegro Valls; Elisabeth Mansur; Maria Lucia Carneiro Vieira
Arachis retusa Krapov. et W. C. Gregory et Valls is endemic in the West-central region of Brazil, occurring in areas endangered by human actions. The establishment of in vitro preservation methods for wild species of Arachis isan alternative to seed banks for germplasm storage, multiplication and distribution. The risk of genetic changesinduced by tissue culture and the monitoring of the genetic stability of the biological material before, during andafter storage must be considered in the context of conservation. Random amplified polymorphic DNA (RAPD) andamplified fragment length polymorphism (AFLP) fingerprinting were used to evaluate the genetic stability of invitro plants originated from cotyledons and embryo axes of A. retusa. Cotyledons originated shoots through directorganogenesis and embryo axes displayed multishoot formation induced by 110 mmol/L and 8.8 mmol/L BAP,respectively. Ninety genomlc regions (loci) generated from RAPD and 372 from AFLP analyses were evaluated. Allamplified fragments detected by both techniques in plants derived from the two explant types were monomorphic.The results indicate that the recovered shoots are genetically stable at the assessed genomic regions.
MARCELA VERSIANI VENÂNCIO PIRES
Full Text Available RESUMO Neste trabalho, objetivou-se avaliar a variabilidade genética da coleção de trabalho de acessos de araticum da Embrapa Cerrados e outros materiais próximos ao Distrito Federal, utilizando marcadores moleculares RAPD, microssatélites e análise de características morfológicas. Folhas de 18 acessos de araticum foram coletadas e utilizadas para a extração das amostras de DNA genômico, as quais foram amplificadas para obtenção de marcadores moleculares RAPD e microssatélites. Na análise morfológica, foram avaliadas 23 características dos acessos de araticum. As dissimilaridades genéticas entre os 18 genótipos de araticum evidenciaram a variabilidade genética dos acessos e as análises de agrupamento levaram à formação de três grupos de similaridade. Verificaram-se coeficientes de dissimilaridades genéticas baixos entre os materiais oriundos da Embrapa Cerrados e altos entre os outros materiais. Esses acessos são importantes fontes de variabilidade para o enriquecimento da atual coleção de trabalho da Embrapa Cerrados e para futuros estudos de caracterização morfológica e agronômica.
Suhrid R. Dutta
Full Text Available The tropical tasar silkworm, Antheraea mylitta, is a semi-domesticated vanya silk-producing insect of high economic importance. To date, no molecular marker associated with cocoon and shell weights has been identified in this species. In this report, we identified a randomly amplified polymorphic DNA (RAPD marker and examined its inheritance, and also developed a stable diagnostic sequence-characterized amplified region (SCAR marker. Silkworms were divided into groups with high (HCSW and low (LCSW cocoon and shell weights, and the F2 progeny of a cross between these two groups were obtained. DNA from these silkworms was screened by PCR using 34 random primers and the resulting RAPD fragments were used for cluster analysis and discriminant function analysis (DFA. The clustering pattern in a UPGMA-based dendogram and DFA clearly distinguished the HCSW and LCSW groups. Multiple regression analysis identified five markers associated with cocoon and shell weights. The marker OPW16(905 bp showed the most significant association with cocoon and shell weights, and its inheritance was confirmed in F2 progeny. Cloning and sequencing of this 905 bp fragment showed 88% identity between its 134 nucleotides and the Bmc-1/Yamato-like retroposon of A. mylitta. This marker was further converted into a diagnostic SCAR marker (SCOPW 16(826 bp. The SCAR marker developed here may be useful in identifying the right parental stock of tasar silkworms for high cocoon and shell weights in breeding programs designed to enhance the productivity of tasar silk.
Full Text Available The karyological and molecular analysis of four fresh water fish species of Family: Poecilidae and their genetic relationships have been studied. Xiphophorus maculates and Xiphophorus hellerii have the same diploid chromosome number 2n=48, but they were different in their karyotypes. Poecilia sphenops and Poecilia reticulata have the same diploid chromosome number 2n=46 and the same fundamental number FN=46, also the same karyotype one group of acrocentric chromosomes. Nine RAPD primers, showed monomorphic bands, were used for the construction of the dendrogram and a similarity matrix. A total of 65 bands were obtained; 39 of them were monomorphic bands. Similarity values among the studied samples ranged from 21% to 38%. High similarity value was obtained between Xiphophorus maculates and Xiphophorus hellerii. (38% and the low similarity values were obtained between Xiphophorus hellerii and Poecilia reticulata (21%. The cluster analysis clearly differentiated Xiphophorus maculates and Xiphophorus hellerii from Poecilia sphenops and Poecilia reticulata. RAPD analysis confirmed that the four species under study are genetically different from each other. These cytogenetic data obtained can be applied for further studies in cytotaxonomy and evolutionary relationships of fishes.
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 O FML (Ligame de Fucose-Manose de Leishmania donovani é uma fração glicoproteica complexa. O seu potencial no diagnóstico da leishmaniose visceral humana foi testado com soros provenientes de Natal, Rio Grande do Norte, Brasil. O teste de FML-ELISA mostrou 100% de sensibilidade e 96% de especificidade, identificando pacientes com calazar declarado (p<0.001, comparados com soros normais e indivíduos com infecção subclínica. Mais de 20% dos sororreativos assimptomáticos desenvolveram a doença no prazo de 10 meses. Na análise de doadores de sangue, 5% de sororeativos, atingindo até 17% num único dia foram detectados. A glicoproteínaGP36 do FHL é reconhecida especificamente por soros de pacientes com calazar. O potencial imunoprotetor do FML no calazar experimental foi testado no modelo swiss albino em combinação com saponina pelas vias subcutâneas e/ou intraperitoneal seguido de desafio com 2x 10(7 amastigolas de Leishmania donovani. Um aumento de 80.0% na resposta de anticorpos específicos (p<0.001 e a redução de 85.5 % da carga parasitária no fígado (p<0.001 foi detectado nos animais vacinados com FML e saponina, independentemente da via de administração.
WANG Li; YANG Juan; GUO Jing; ZHAO Guifang
Psathyrostachys huashanica Keng is a perennial grass and belongs to genus Psathyrostachys under Triticeae.sathyrostachys is found in the center of Middle Asia and the Caucasus Mountain,while P.huashanica,a species endemic to China,is only located in Mt.Hua in the Shaanxi province,China.At present,the population of this species is decreasing,and reaching the edge of extinction.Due to the limitation in distribution and the importance as breeding material for germplasm storage,it has been considered as first class among the national protected rare plants.For this reason,the present study is significant in probing plant flora,origin and evolution of Triticeae,and crop breeding.Randomly amplified polymorphic DNA (RAPD)markers were used to analyze the genetic structure and differentiation of P.huashanica populations sampled in three valleys (Huangpu,Xian,and Huashan Valleys)in Mr.Hua.One hundred and twenty-two RAPDfragments were obtained in all 266 individuals with 20 primers with a mean of 6.1 (2-10)fragments per primer.The percentage of polymorphic loci (PPB)was 60.66%in Huangpu Valley,90.98%in Xian Valley,95.08%in Huashan Valley,and the total PPB was 95.08%,which indicated a highly genetic variability of P.huashanica.The Shannon's Information index and GST were 0.3306 and 0.3263,respectively,indicating that there were more genetic variations within the subpopulations than those among the subpopulations.The gene flow among the subpopulations of P.huashanica (Nm=1.0322)was much less than that of the common anemophytes (Nm=5.24).Mean genetic distance is 0.1571(range:0.0022-0.2901).The highest value of genetic distance was found between the subpopulation (hp1)of Huangpu Valley and the highest altitude subpopulation (h8)of Huashan Valley.Correlation analysis detected significant correlation between genetic distance and vertical distance of altitude.Clustering analysis and principal coordinate analysis revealed the genetic differentiation among the populations of P
Frello, S.; Hansen, K.R.; Jensen, J.;
We have examined the inheritance of 20 rapeseed (Brassica napus)-specific RAPD (randomly amplified polymorphic DNA) markers from transgenic, herbicide-tolerant rapeseed in 54 plants of the BC1 generation from the cross B. juncea x (B. juncea x B. napus). Hybridization between B. juncea and B. napus...
Full Text Available The aim of this study was to assess the genotoxic effects of environmental chemicals on residents living near landfills. The study was based on samples of amniotic fluid from women living in the intensely polluted areas around the Campania region of Italy compared to a nonexposed control group. We evaluated the genetic effects that this amniotic fluids collected in contaminated sites had on Paracentrotus lividus embryos. DNA damage was detected through changes in RAPD (Random Amplified Polymorphism DNA profiles. The absence of the amplified DNA fragments indicated deletions in Paracentrotus lividus DNA exposed to the contaminated amniotic fluids when compared to equal exposure to uncontaminated fluids. These results show the ability of RAPD-PCR to detect and isolate DNA sequences representing genetic alterations induced in P. lividus embryos. Using this method, we identified two candidate target regions for DNA alterations in the genome of P. lividus. Our research indicates that RAPD-PCR in P. lividus embryo DNA can provide a molecular approach for studying DNA damage from pollutants that can impact human health. To our knowledge, this is the first time that assessment of DNA damage in P. lividus embryos has been tested using the RAPD strategy after exposure to amniotic fluid from residents near waste landfill sites.
Dasgupta, Nirjhar; Nandy, Paramita; Sengupta, Chandan; Das, Sauren
Increased salinity distresses some key species severely in Indian Sundarbans. Geomorphic characteristics coupled with demographic obligations have proven to be pivotal factor towards the prevalence of elevated salinity in this zone. Better adaptation to rapid changes in microclimate demands wide range of genetic polymorphism as well. RAPD and ISSR molecular markers were used for this genetic diversity study. Degree of polymorphism was found relatively higher in Bruguiera gymnorrhiza (26.43 % in RAPD and 24.36 % in ISSR) than the other taxa, Heritiera fomes (14.43 and 12.76 % respectively) in case of RAPD and ISSR. Dendrogram constructed based on the similarity matrix showed that for H. fomes, least saline and highest saline zones are positioned in the same clade; whereas in B. gymnorrhiza the higher saline areas were clustered together. Nei's gene diversity (h) as revealed from RAPD and ISSR analysis were found to be 0.0821, 0.0785 and 0.0647, 0.0592 in B. gymnorrhiza and H. fomes respectively. The higher degree of polymorphism as revealed from UPGMA Dendrogram and Nei's genetic diversity might be attributed towards the comfortable growth of B. gymnorrhiza all along the Indian Sundarbans. On the other hand the relatively lesser degree of genetic polymorphism of H. fomes might be attributed towards their precarious status in present days elevated salinity in Indian Sundarbans. PMID:26261402
Pamela E. Akin-Idowu
Full Text Available Efficient utilization of plant genetic resources for nutrition and crop improvement requires systematic understanding of the important traits. Amaranthus species are distributed worldwide with an interesting diversity of landraces and cultivars whose leaves and seeds are consumed. Despite their potential to enhance food security and economic livelihoods, grain amaranth breeding to improve nutritional quality and adoption by farmers in sub-Saharan Africa is scanty. This study assessed the variation among 29 grain amaranth accessions using 27 phenotypic (10 morphological and 17 nutritional characters and 16 random amplified polymorphic DNA (RAPD primers. Multivariate analysis of phenotypic characters showed the first four principal components contributing 57.53% of observed variability, while cluster analysis yielded five groups at 87.5% similarity coefficient. RAPD primers generated a total of 193 amplicons with an average of 12.06 amplicons per primer, 81% of which were polymorphic. Genetic similarities based on Jaccard’s coefficient ranged from 0.61 to 0.88. The RAPD-based unweighted pair group method with arithmetic mean dendrogram grouped the accessions into nine clusters, with the same species clustering together. RAPD primers distinguished the accessions more effectively than phenotypic markers. Accessions in the different clusters as obtained can be exploited for heterotic gain in desired nutritional traits.
Atienzar, Franck A; Jha, Awadhesh N
The random amplified polymorphic DNA (RAPD) is a useful assay for the detection of genotoxin-induced DNA damage and mutations. In this study, we have further evaluated the potential of this assay to measure benzo(a)pyrene [B(a)P]-induced DNA changes, and repair (in kinetic experiments) as well as transgenerational effects in the water fleas, Daphnia magna. The organisms, which reproduce parthenogenetically, were exposed to 50 microg L(-1) B(a)P for 3 or 6 days and were allowed to recover in clean medium for 12 or 9 days, respectively. Qualitative and quantitative changes were observed in RAPD profiles generated not only from the B(a)P exposed Daphnia but also from previously treated organisms during the recovery experiments. The fact that some of the RAPD changes disappeared at the end of both recovery experiments suggested that the DNA effects were fully repaired or reversed. In addition, some of the B(a)P-induced RAPD alterations detected in parental D. magna were also observed in the offspring patterns. This suggested that DNA alterations that occurred in germ cells were probably transmitted to the next cohorts. The present study shows that the RAPD method can be useful to qualitatively assess the kinetics of DNA changes, repair and transgenerational effects and such effects could potentially be linked to survival and reproductive success at higher levels of biological organisation. In addition, the water fleas have efficient capabilities to repair or reverse B(a)P-induced DNA effects. Finally, unrepaired or misrepaired genetic damage induced by genotoxins such as B(a)P could be transmitted to next generations in these parthenogenetically reproducing organisms. PMID:15288546
Random Amplified Polymorphic DNA (RAPDs) markers were utilized to detect polymorphism between pure lines and commercially available Basmati rice varieties to assess variation which may be helpful in quality control and varietal identification (Basmati-370 and derived radiation induced mutants), differentiation of mutants and parents, and identification of RAPD markers co-segregating with important agronomic traits including plant height, days to flower and grain quality. Basmati varieties were distinguished from non-Basmati varieties with the help of five diagnostic markers which will be useful for detecting mixing of non-Basmati and Basmati rices, currently a serious marketing problem. Different Basmati cultivars were identified with the help of diagnostic RAPD markers which can be used in quality control as well as for ''fingerprinting'' of cultivars. Different radiation induced mutants were also successfully distinguished from the parents on the basis of variety specific and mutant specific markers which will be useful for varietal identification. In addition to this, other markers were also identified which can differentiate mutants from each other and are being, used for the fingerprinting of different mutants, particularly the dwarf mutants having similar appearance but different parentage. For identification of RAPD markers co-segregating with plant height and days to flower, 50 F2 plants and four F3 families were studied from a reciprocal cross made between Kashmir Basmati (tall and early) and Basmati-198 (dwarf and late). Segregating bands were observed within these populations, and indicating the possible use of RAPD markers for tagging gene(s) of agronomic importance in rice. (author)
The koinobiont endoparasitoid Opius concolor (Spzel.) has been used as a biological control agent in area-wide biological control programmes of the olive fruit fly Bactrocera oleae (Gmelin), which consists of massive releases of parasitoids in fields where indigenous wild ones are present. Field differentiation between the released and the indigenous parasitoids, and the estimation of the proportion of released individuals in the field population are a prerequisite to estimate the efficiency of biological control programmes. Because wild and released parasitoids are morphologically indistinguishable, a DNA molecular technique, based on Polymerase Chain Reaction (PCR), the Random Amplified Polymorphic DNA (RAPD-PCR) was used to differentiate between five different populations of this parasitoid: Two populations from Jordan, two populations from southern Italy and a laboratory reared strain. RAPD-PCR is a powerful and useful technique when there is no prior knowledge of the genome and the DNA sequence of the tested organism as in the case of the parasitoid O. concolor. In RAPD-PCR three different primers (NP4, D17, A10) were used to test polymorphism between individuals from the five populations by checking the presence/absence of amplified DNA bands from the PCR reactions. Where certain bands were found in some individuals and not found in others, those bands are called polymorphic bands and they could be used to distinguish between individuals within the same population or individuals of one population and individuals of another population. The genomic DNA amplification of 90 individuals of O. concolor, from five different populations (18 individuals from each), two from Jordan, two from southern Italy and the laboratory strain, using the three primers (A10, D17, NP4), resulted in a total of 39 interpretable bands ranging in size from 170 bp to 830 bp. Twenty-one of these bands were polymorphic (i.e. present in some individuals and absent in others). Two strain
Ali Saeed Atiyah AL-Janabi
Full Text Available Sweet orange (Citrus sinensis L. Osbeck is one of the most important commercially cultivated fruit crops of Citrus. Genetic diversity and inter-relationship among 5 cultivars (Indian, Iraqi, Japanese, Syrian, Egyptian of C. sinensis were analyzed based on RAPD markers. Six primers generated reproducible and easily storable RAPD profiles with a number of amplified DNA fragments ranging from 6 to 14 fragment bands. The total number of amplicons detected was 51, including 14 fragments unique bands with average reached 2.8 fragments ̸ primers. While the number of polymorphic ranged from 0 to 8 with an average reached 4.4 fragments ̸ primers with the polymorphic percentage ranged from 0% to 57.1%. While the number of monomorphic ranged from 2 to 5 fragment bands and was total of the monomorphic 15 fragments with an average reached 3 fragments ̸ primers with the monomorphic percentage was 14.2 % to 83.3%. A maximum numbers of amplicons was amplified with primer OPS-238 reached 14 fragments while the minimum number of fragments was amplified with primer OPS-253 reached 6 fragments. The highest number of polymorphic bands reached 8 fragments was obtained with primer OPS-238 with high percentage 57.1%, while the highest number of monomorphic bands reached 5 fragments with high percentage 83.3% was obtained with primer OPS-253. RAPD markers detected genetic distance and similarity, amaximum genetic distance value was observed between Japanese (Jap and Syrian (Syr cultivars reached 0.530 with less similarity value reached 47%, a minimum genetic distance value was observed between sweet Iraqi (Irq and Indian (Ind cultivars reached 0.239 with high similarity value reached 76.1%. The similarity matrices were employed in the cluster analysis to generate a dendrogram using the UPGMA method. The cluster tree analysis showed that the sweet orange cultivars were broadly divided into two main groups A and B with similarity reached 50%. A group including
Full Text Available O objetivo deste trabalho foi a caracterização genética da raça bovina Crioulo Lageano por marcadores RAPD em comparação com as raças Holandesa e Nelore. Foram selecionados 43 primers, que geraram 77 bandas polimórficas. Os animais foram distribuídos em cinco subgrupos de Crioulo Lageano (I a V, e um subgrupo em cada uma das raças Holandesa (VI e Nelore (VII. A maior parte da variância genética total (65,05% foi causada pela diferença de indivíduos dentro dos grupos, e o restante pelas diferenças entre grupos. A análise conjunta dos grupos I a V apresentou variabilidade genética entre grupos de 25,28% e dentro dos grupos de 74,72%. A diversidade gênica vem se mantendo ao longo das gerações no núcleo de conservação do Crioulo Lageano. A raça Holandesa apresentou a menor diversidade gênica (0,1204, e a Crioulo Lageano a maior (0,3154. A maior distância genética (0,3747 foi entre as raças Nelore e Holandesa. Os grupos de Crioulo Lageano apresentaram diferenças entre si e apenas alguns indivíduos de cada grupo posicionaram-se junto a outros grupos. A técnica RAPD é capaz de estimar a distância genética entre raças ou populações e de auxiliar na escolha de indivíduos, visando aos trabalhos de conservação de recursos genéticos.The objective of this study was to characterize genetically the Crioulo Lageano cattle breed, using RAPD markers and compare it to the Holstein and Nelore breeds. Forty three primers were selected, and they generated 77 polymorphic bands. Seven groups were studied: 5 subgroups of Crioulo Lageano (I to V and one each Holstein (VI and Nellore (VII. Using all groups, the greater part of the genetic variance (65.05% was due to differences within groups and the rest due to differences between groups. Using five Crioulo Lageano groups (I to V the results showed 25.28% variation between groups and 74.72% within groups. Genetic diversity has been maintained throughout the generations in this
Full Text Available Em programas de melhoramento de citros, a caracterização adequada dos recursos genéticos disponíveis é de grande importância, principalmente devido às características biológicas da cultura, como a heterozigosidade, a embrionia nucelar e o longo ciclo reprodutivo. A facilidade com que ocorrem hibridações (interespecíficas e intergenéricas e a embrionia nucelar favoreceram a formação e a preservação de novas combinações, classificadas como espécies. Neste estudo, marcadores RAPDs foram utilizados para analisar 15 acessos de Citrus spp., sendo quatro variedades de laranjeiras doce (C. sinensis Osbeck, quatro tangerineiras (C. reticulata Blanco, C. nobilis Loureiro, C. sunki Loureiro e C. deliciosa Tenore, uma laranjeira azeda (C. aurantium L., um pomeleiro (C. paradisi Macf., uma torangeira (C. grandis Osbeck, uma cidreira (C. medica L., uma limeira ácida (C. latifolia e dois híbridos (Citrus clementina T. x (C. tangerina T. x C. paradisi Macf.. Doze sequências iniciadoras aleatórias foram utilizadas para estudar os 15 genótipos, encontrando-se um grau de similaridade mínimio de 0,81 ("Simple Matching" entre as tangerineiras. Os menores graus de similaridade foram encontrados entre as espécies de Citrus menos aparentadas (C. medica, C. grandis e C. latifolia. As quatro cultivares de laranjeiras doces não puderam ser diferenciadas pelos marcadores RAPD utilizados, apresentando similaridade máxima.In citrus improvement programs the characterization of the available genetic resources is of great importance, mainly concerning biological characteristics of the culture, as the heterozigosity, nucellar the embriony and long reproductive cycle. Favored by nucellar embriony interespecific and intergeneric hybridizations and genotypes preservation happen easily. RAPDs markers were used to analyze 15 Citrus spp., four sweet orange (C. sinensis Osbeck, (C. medica, C. grandis e C. latifolia, four mandarins (C. reticulata Blanco, C
Ana Maria Barros
Full Text Available Stylosanthes macrocephala M. B. Ferr. et S. Costa é uma leguminosa utilizada sob consorciação em pastagens, adubação e recuperação de áreas degradadas. A falta de características morfológicas e agronômicas estáveis e de informações ecogeográficas dos locais de coleta dos acessos tem dificultado o melhoramento genético da espécie. A fim de obter descritores ecológicos, moleculares e avaliar a variabilidade genética da coleção de S. macrocephala, 87 acessos foram analisados com o auxílio do Sistema de Informações Geográficas (SIG e de marcadores moleculares RAPD. Os acessos provieram de sete Estados, cinco bacias hidrográficas, sete tipos de vegetação e sete tipos de solos. As altitudes dos locais de coleta variaram de 1 a 1.298 m e a pluviometria anual média de 550 a 2.870 mm. A variabilidade de descritores ecológicos sugeriu diversidade adaptativa na coleção. Com base em 161 marcadores RAPD, verificou-se que as distâncias genéticas entre os acessos de S. macrocephala variaram entre 0,02 e 0,42. Com base nessas distâncias, dez grupos de similaridade genética foram estabelecidos. Observou-se tendência de separação por bacias hidrográficas e elevada variabilidade gen��tica entre os acessos coletados nos estados da Bahia e de Minas Gerais. A alta variabilidade genética da coleção de S. macrocephala evidencia a importância desses acessos para futuros trabalhos de melhoramento genético.Stylosanthes macrocephala M. B. Ferr et S. Costa is a leguminous species used as forage, cover crop and as a pioneer plant to recover degraded areas. Inexistence of stable morphological descriptors and lack of ecogeographic information about collecting sites bring difficulties to the studies of this species. The objective of this work was to use the geographic information system (GIS and RAPD markers to obtain ecological and molecular descriptors and to study the genetic variability of 87 S. macrocephala accessions. The
郑敏; 罗玉萍; 李思光; 朱必凤
以南丰蜜桔树叶为基因组DNA提取材料,建立了其RAPD-PCR分析的最佳反应体系:模板DNA1.5ng/μl,随机引物0.6μM@L-1,dNTPs各0.1mM@L-1,MgCl2.0mM@L-1,BSA 250ng/μl,Tris-HCl 10mM@L-1,pH9.0,KCl 50mM@L-1,Nonidet P400.1%,Taq酶0.5u,总反应体积20μl.
Determinación de la variabilidad genética entre aislamientos de Rosellinia sp. Rosellinia bunodes y Rosellinia pepo mediante la técnica de amplificación aleatoria de polimorfismos de DNA (RAPD y análisis de los espaciadores de transcritos internos (ITSS
Ángel Díaz Jorge Evelio
Full Text Available Teniendo en cuenta que las medidas preventivas son las más eficientes para reducir la incidencia de muchas enfermedades, la implementación de estrategias que permitan el diagnóstico temprano y oportuno de fitopatógenos de cultivos, en especial aquellos de tardío rendimiento, es el objetivo principal de este trabajo. Los hongos fitopatógenos del género Rosellinia se consideran habitantes naturales del suelo. Sin embargo, su fase parasítica se asocia a la aparición de enfermedades radiculares principalmente en cultivos de café, papa y cacao, que generalmente se
relaciona con el aumento en la oferta de materia orgánica, lo cual beneficia el crecimiento sus poblaciones y las de otros microorganismos que pueden ser patógenos. Con el fin de avanzar en el conocimiento de las especies Rosellinia sp., Rosellinia bunodes y Rosellinia pepo y de desarrollar estrategias de diagnóstico para evitar las pudriciones radiculares que estas ocasionan en cultivos de importancia agroeconómica y forestal, se utilizaron dos metodologías moleculares que permiten
realizar un primer acercamiento a la variabilidad genética que presentan diferentes aislamientos de estas especies. Mediante el análisis de RAPD (Amplificación Aleatoria de Polimorfismos de ADN y la secuenciación de las regiones de rDNA-ITS (Espaciadores de Transcritos Internos fue posible establecer un alto grado de variabilidad entre las cepas, aún dentro de una misma especie. Dichas metodologías están basadas en la reacción en cadena de la polimerasa (PCR que ha sido extensamente utilizada tanto en la identificación de un organismo por medio de la amplificación selectiva de su ADN, como al ser aplicada para explorar la variabilidad que posee un genoma. Las cepas de Rosellinia sp. presentaron gran variación en los perfiles electroforéticos de RAPD constituyendo dos ramas aisladas en el dendrograma de similaridad construido a partir de matrices binarias. De la misma manera
HUGO BRUNO MOLINARI
Full Text Available A identificação e caracterização da diversidade genética de plantas por meio de técnicas moleculares envolvem a avaliação de vários indivíduos, necessitando-se, portanto, de métodos rápidos e precisos de extração do DNA. O co-isolamento de polissacarídeos, fenóis e compostos secundários é o principal problema encontrado no isolamento e purificação de DNA vegetal. Folhas das diversas espécies de Passiflora possuem níveis variados desses compostos que podem comprometer este procedimento. O presente estudo foi realizado com o objetivo de avaliar a qualidade e quantidade de DNA de folhas de variedades de Passiflora spp., utilizando-se de três métodos de extração. Os três métodos forneceram DNA em qualidade e quantidade suficientes para a realização da técnica PCR-RAPD.The identification and characterization of the genetic diversity of plants by molecular techniques involve the evaluation of several individuals, therefore requiring fast and precise extraction methods of DNA. Co-isolation of polysaccharides, phenols and secondary products is the main problem during isolation and purification of plant DNA. The leaves of several species of Passiflora have different levels of those compounds that can compromise this procedure. The objective of this study was to evaluate the quality and amount of DNA extracted from Passiflora spp. varieties using three extraction methods. The three methods supplied DNA in quality and quantity sufficient amount for PCR-RAPD analyses.
José Maria D. Gaia
Full Text Available A espécie conhecida como pimenta-de-macaco (Piper aduncum L. possui grande potencial para exploração econômica em função da comprovada utilidade do seu óleo essencial na agricultura e saúde humana. O objetivo deste trabalho foi caracterizar a diversidade genética de populações naturais dessa planta. Um total de dezoito acessos da planta, provenientes de quatro procedências da Amazônia Brasileira, foi examinado por meio de locos de DNA, gerados por análise RAPD (polimorfismo de DNA amplificado ao acaso. O estudo evidenciou a existência de real diversidade entre as populações examinadas, sendo provável que dentro das localidades investigadas, os padrões da diversidade genética acompanhem os padrões de distribuição geográfica.The species known as pimenta-de-macaco (Piper aduncum L. has great economic explotation potential based on the proved usefulness of the essential oil in agriculture and human health. The genetic diversity of their natural populations was characterized. A total of eighteen accessions of the plant, obtained from four different origins in the Brazilian Amazon, was examined by means of DNA loci, generated by RAPD analysis. Real genetic diversity was observed between the analyzed populations and it appear that the patterns of the genetic diversity follow the patterns of the geographical distribution.
Gebre-Michael, T; Malone, J B; Balkew, M; Ali, A; Berhe, N; Hailu, A; Herzi, A A
The distribution of two principal vectors of kala-azar in East Africa, Phlebotomus martini and Phlebotomus orientalis were analysed using geographic information system (GIS) based on (1) earth observing satellite sensor data: Normalized Difference Vegetation Index (NDVI) and midday Land Surface Temperature (LST) derived from advanced very high resolution radiometer (AVHRR) of the global land 1km project of United States Geological Survey (USGS), (2) agroclimatic data from the FAO Crop Production System Zone (CPSZ) of the Intergovernmental Authority on Development (IGAD) sub-region, and (3) the FAO 1998 soils digital map for the IGAD sub-region. The aim was to produce a predictive risk model for the two vectors. Data used for the analysis were based on presence and absence of the two species from previous survey collections in the region (mainly Ethiopia, Kenya and Somalia). Annual, wet season and dry season models were constructed. Although all models resulted in more than 85% positive predictive values for both species, the best fit for the distribution of P. martini was the dry season composite (NDVI 0.07-0.38 and LST 22-33 degrees C) with a predictive value of 93.8%, and the best fit for P. orientalis was the wet season composite (NDVI -0.01 to 0.34 and LST 23-34 degrees C) with a predictive value of 96.3%. The two seasonal composites models derived from satellite data were largely similar with best fit models developed based on the CPSZ climate data: average altitude (12-1900m), average annual mean temperature (15-30 degrees C), annual rainfall (274-1212mm), average annual potential evapotranspiration (1264-1938mm) and readily available soil moisture (62-113mm) for P. martini; and average altitude (200-2200m), annual rainfall (180-1050mm), annual mean temperature (16-36 degrees C) and readily available soil moisture (67-108mm) for P. orientalis. Logistic regression analysis indicated LST dry season composite of the satellite data, average altitude, mean annual
Jeyaram, A.; Kesari, S.; Bajpai, A.; Bhunia, G. S.; Krishna Murthy, Y. V. N.
Visceral Leishmaniasis (VL) commonly known as Kala-azar is one of the most neglected tropical disease affecting approximately 200 million poorest populations 'at risk in 109 districts of three endemic countries namely Bangladesh, India and Nepal at different levels. This tropical disease is caused by the protozoan parasite Leishmania donovani and transmitted by female Phlebotomus argentipes sand flies. The analysis of disease dynamics indicate the periodicity at seasonal and inter-annual temporal scale which forms the basis for development of advanced early warning system. Study area of highly endemic Vaishali district, Bihar, India has been taken for model development. A Systematic study of geo-environmental parameters derived from satellite data in conjunction with ground intelligence enabled modelling of infectious disease and risk villages. High resolution Indian satellites data of IRS LISS IV (multi-spectral) and Cartosat-1 (Pan) have been used for studying environmentally risk parameters viz. peri-domestic vegetation, dwelling condition, wetland ecosystem, cropping pattern, Normalised Difference Vegetation Index (NDVI), detailed land use etc towards risk assessment. Univariate analysis of the relationship between vector density and various land cover categories and climatic variables suggested that all the variables are significantly correlated. Using the significantly correlated variables with vector density, a seasonal multivariate regression model has been carried out incorporating geo-environmental parameters, climate variables and seasonal time series disease parameters. Linear and non-linear models have been applied for periodicity and interannual temporal scale to predict Man-hour-density (MHD) and 'out-of-fit' data set used for validating the model with reasonable accuracy. To improve the MHD predictive approach, fuzzy model has also been incorporated in GIS environment combining spatial geo-environmental and climatic variables using fuzzy membership
Full Text Available Genotype or cultivar identification is the first step to ensure plant breeder’s rights. If registered cultivars or germplasms were not distinguished by any mechanism, then seed production or novel varieties could not be controlled. The RAPD markers are chosen to distinguish seven soybean ( Glycine max. L. Merrill cultivars from each other and to assign the polymorphisms among of them in this study. Eight primers were used and DNA fingerprinting of all cultivars were determined with that primers. According to that fingerprinting all cultivars were distingusihed from each other in DNA banding pattern. Using these markers results, similarity index in between soybean cultivars were calculated. Dwight and Maverick were the most similar cultivars ( These two cultivars were similar at 0,70 point to each other. In addition that Maverick and Defiance cultivars have shown the highest polymorphism rate ( These two cultivars were similar at 0,39 point each other. .
A stable cms line named D-type cms line was obtained after injection of exogenous λ DNA into wheat line 814527. Line 812547 can be its maintainer. In order to detect the differences on molecular level among λ DNA, receptor 814527, sterile line and its F1 hybrid, ctDNA RAPD analysis was made. The results showed that one specific band existed in the donor of the sterile line and its F1 hybrid, but not existed in the receptor. Two specific bands existed in the receptor and F1 hybrid, but disappeared in the sterile line. These bands might be related to male sterility. It also proved that the DNA fragment of the donor could be integrated into the genome of receptor, which might disorder the expressing and controlling of the genes, and then bring about mutations
Aline R. Rabello
Full Text Available The Bemisia tabaci complex is formed by approximately 41 biotypes, two of which (B and BR occur in Brazil. In this work we aimed at obtaining genetic markers to assess the genetic diversity of the different biotypes. In order to do that we analyzed Bemisia tabaci biotypes B, BR, Q and Cassava using molecular techniques including RAPD, PCR-RFLP and sequencing of the ITS1 rDNA region. The analyses revealed a high similarity between the individuals of the B and Q biotypes, which could be distinguished from the BR individuals. A phylogenetic tree based on ITS1 rDNA sequence was constructed. This is the first report of the ITS1 rDNA sequence of Bemisia tuberculata and of the BR biotype of B. tabaci.
Saini, A; Dua, A; Mohindra, V
The Giant river catfish, Sperata seenghala (Sykes) is commercially very important fish species of South Asia. Genetic variability between its populations collected from two rivers i.e. river Sutlej and river Beas of Indus river system in India were examined using randomly amplified polymorphic DNA analysis. Total 38 fish samples were collected from river Sutlej whereas 46 fish samples were collected from river Beas. Total 40 primers were screened, out of these 7 were selected for studying polymorphism which produced a total of 64 RAPD loci in two populations. Percentage polymorphic loci calculated following 95% criterion was 89.06% for Beas population as compared to 95.31% for Sutlej population. Moderate level of genetic divergence (genetic distance of 0.0486) between both the populations suggests distinct population substructure of giant river catfish in both the rivers. PMID:20873207
YI HuaLin; DENG XiuXin
Huazhong Agricultural University, Wuhan 430070, ChinaThirty-one polymorphic decamer primers were selected to genotype 92 progenies from the cross between Yiben No,4, a monoembryonic diploid F1 hybrid of Citrus reticulata Blanco cv Huanongbendizao tangerine and C. ichangensis Swingle, and [Hamlin sweet orange + Rough lemon], an allotetraploid somatic hybrid of Citrus sinensis Osbeck cv. Hamlin and C. jambhiri Lush cv. Rough Lemon. x2(Chi-square) analysis of RAPD markers in the progenies indicated they were randomly transmitted from the four donor parents, without significant difference between the diploids and triploids. However,these progenies were clustered into three major groups using dendrogram constructed by UPGMA,skewed to three parents in certain degrees, 15 (13 triploids and 2 diploids) to Hamlin, 16 (9 and 7) to Yiben No. 4, and 61 (57 and 4) to [Hamlin sweet orange + Rough Lemon] from which genomic contribution was predominant in progenies, respectively.
Thirty-one polymorphic decamer primers were selected to genotype 92 progenies from the cross be- tween Yiben No.4, a monoembryonic diploid F1 hybrid of Citrus reticulata Blanco cv Huanongbendizao tangerine and C. ichangensis Swingle, and [Hamlin sweet orange + Rough lemon], an allotetraploid somatic hybrid of Citrus sinensis Osbeck cv. Hamlin and C. jambhiri Lush cv. Rough Lemon. χ2 (Chi-square) analysis of RAPD markers in the progenies indicated they were randomly transmitted from the four donor parents, without significant difference between the diploids and triploids. However, these progenies were clustered into three major groups using dendrogram constructed by UPGMA, skewed to three parents in certain degrees, 15 (13 triploids and 2 diploids) to Hamlin, 16 (9 and 7) to Yiben No. 4, and 61 (57 and 4) to [Hamlin sweet orange + Rough Lemon] from which genomic contribu- tion was predominant in progenies, respectively.
Adriana Daniela BACIU
Full Text Available In order to identify cultivars with special decorative value and potential genitors for breeding process, 45 genotypes of Calendula genera were analyzed, belonging to six species: C. officinalis, C. alata, C. arvensis, C. stellata, C. suffruticosa and C. tripterocarpa. The average height of plants varied strongly, from 22.0 cm (C. officinalis cv. Rozovyi Sjurpriz cm to 84.1 cm (C. tripterocarpa Rupr.. The lowest number of branches per plant was recorded on C. arvensis F. (4.6 and the largest one on C. officinalis LDA (16.4. The average number of flowers per plant ranged from 98.0 (C. suffruticosa Valh. to 2.0 (C. officinalis UK. From among all genotypes, aphids (Aphis fabae have attacked 19 (42.2%, and the results showed that AD% (Attack Degree depend significantly on genotypes. RAPD analysis and phylogenetic dendrogram illustrated the relationship between genotypes and DNA polymorphism exists between the six species. Were found not only close phylogenetic links among cultivars apart of the same specie, but also between different species. C. officinalis A., C. alata UK and C. suffruticosa formed a subgroup similar to the molecular level, but also confirming some phenotypic similarities, these species having the smallest number of petals in the corolla and the highest sensitivity to Aphis fabae attack. The large variability identified in Calendula genotypes allows the selection of potential genitors for new breeding works, with appropriate decorative characteristics and resistance to aphids attack. RAPD analyses and phenotypic study allows hypothesis regarding the success of intra- and inter-specific hybridization, thus facilitating Calendula breeding processes.
Lanzone, Valentina; Tofalo, Rosanna; Fasoli, Giuseppe; Perpetuini, Giorgia; Suzzi, Giovanna; Sergi, Manuel; Corrado, Federica; Compagnone, Dario
Random amplified polymorphic DNA (RAPD) PCR is a feasible method to evaluate genotoxin-induced DNA damage and mutations. In this study, Lactobacillus plantarum ATCC 14917T, Enterococcus faecium DSMZ 20477T, Escherichia coli PQ37 and Saccharomyces cerevisiae S441 were screened for DNA genetic alterations by DNA fingerprinting using M13 and LA1 primers after treatment with three compounds forming covalent adducts with DNA [benzo[a]pyrenediol epoxide (BPDE), methyl methanesulfonate and 1,2,3,4-diepoxybutane (DEB)]. M13 RAPD fingerprinting revealed that the total number of bands decreased in all treated DNA compared to control samples and generally the lost bands were characterized by high molecular weight. Some extra bands were detected for L. plantarum and E. faecium, while in E. coli and S. cerevisiae DNAs BPDE and DEB treatments did not result in new extra bands. Besides qualitatively analysis, cluster analysis based on Unweighted Pair-Group Method with Average algorithm was performed to compare DNA fingerprints before and after treatments. This analysis confirmed the absence of significant differences between negative controls and treated DNA in S. cerevisiae and E. coli however the disappearance of some bands can be detected. The data indicate that this approach can be used for DNA damage detection and mutations induced by genotoxic compounds and highlighted the possible use of L. plantarum and E. faecium M13 based fingerprinting as reference for hazard identification in risk assessment. PMID:26991971
Wheat is the major winter cereal crop in the world. The total cultivated area of this crop in Egypt is about two million feddans. Soil salinity represent a serious problem to agriculture in arid and semi-arid in the world. Mexico wheat (Triticum vulgar var. Ycora rojo) was imported in 1999 for cultivation. Mexico wheat was exposed to gamma rays (cobalt 60) from 10 to 80 Krad The unirradiated and irradiated wheat were cultivated in the presence of 0, 5000,10000 and 20000 mg/L of salt solution and 16 hour light /25 degree C. The previous treatment was repeated in combination with 5, 10 mg/l ABA and 10, 20 mg/l GA3 separately. Different accessed parameters were used for evaluation, these parameters were: germination percentage, length of shoots and roots, pigment contents (chl. a,b and a/b carotenoids and total pigments), total protein patterns and RAPD, PCR techniques. The results showed that both of radiation and salinity reduced the percentage of germination. Soaking grains in GA3 considerably increased the shoot and root lengths. Highest value of carotenoids obtained act as a defense mechanism against harmful salinity action. Also, the seedling exposed to 80 Krad and treated with ABA (5 or 10 mg/l) can survive during the experimental period, while plants treated with 10 and 20 mg/l GA3 and exposed to 80 Krad can not survive. At low radiation doses (10 and 20 Krad) there was no difference in the number and density of bands of the total protein patterns, while in the RAPD, PCR technique in presence and/or absence of DNA band in unirradiated and irradiated wheat seeds were observed
I Wayan Suardana
Full Text Available Epidemiological studies of zoonotic agent Escherichia coli O157:H7 have been analyzed pheneticallyand or phylogenetically. In a phenetic classification, micoorganisms are arranged into groups (phena onthe basis of high overall similarity using both phenotypic and genotypic methods without judgementaspect of its ancestry or evolutionary. Due to its importance to epidemiological aspect, the study of geneticvariation of isolates origin from some sources need to be conducted in order to trace the routes of infection.A total of 20 samples obtained from some sources i.e clinically human feces, non-clinically human feces,cattle feces, chicken feces, and beef feces were used in this study. The study was started by confirming allof the isolates using O157 latex agglutination test and H7 antiserum test, followed by genomic DNAanalysis by random amplification of polymorphic DNA /RAPD methods. RAPD results were analyzed using a simple matching coeficient (Ssm and alogorhythm unweighted pair group method using arithmeticaverages (UPGMA programe. Results showed there were range of genetic DNA from local isolates (75.1–99,6% which was almost similar to ATCC 43894 control isolate. The highest similarity (99.6% to ATCC43894 control was showed by SM-7(1 isolate obtained from cattle fecal and KL-68(1, isolate obtainedfrom clinically human fecal. In addition, KL-52(7 obtained from clinically human fecal had high similarity(99.6% to MK-35 isolate obtained from chicken fecal. On the other hand, DS-21(4 and DS-16(2 isolatesthat were obtained from beef had high similarity (84.9% to other isolates including ATCC 43894 controlisolate. The highest similarity of E. coli O157:H7 isolates that were obtained from cattle feces, beef, andchicken feces to human feces isolate indicated that there were both cattle and chicken were potentialreservoirs of the zoonotic agen which can be transmitted to human.
Oscar A. Villarreal-Espino-Barros; Fernando X. Plata-Pérez; Germán D. Mendoza-Martínez; José A. Martínez-García; Hernández-García, Pedro A.; José L. Arcos-García
El efecto de la distancia radial al agua (DRA) sobre el volumen vegetal, la cobertura de escape (CE) y la presencia del venado cola blanca, se evaluó. El trabajo se realizó en el estado de Puebla, México. Cinco áreas con una DRA menor a 800 m, cuatro entre 800-1,600 y cuatro mayores a 1,600 m, fueron seleccionadas utilizando un modelo completamente al azar. El volumen vegetal se estimó en estas áreas y fueron el punto de inicio de transectos en los cuales se estimó la CE y el número de grupos...
Alexandre Wagner Silva Hilsdorf; Sara Lívia da Silva Fernandes da Matta; Maria Santina de Castro Morini
Random amplified polymorphic DNA (RAPD) markers were used to investigate the genetic relationship among nests of the carpenter ant, Camponotus rufipes, located in the same area. Five random oligodecamers were used to amplify DNA from 108 ant workers collected from six nests. A total of 47 RAPD markers were identified, which revealed low levels of genetic differentiation among nests (Fst = 0.00218) and a low average Shannon index (0.3727) among workers within nests. These results together sugg...
Preety Panwar; Manoj Nath; Vijay Kumar Yadav; Anil Kumar
Genetic relationships among 52 Eleusine coracana (finger millet) genotypes collected from different districts of Uttarakhand were investigated by using randomly amplified polymorphic DNA (RAPD), simple sequence repeat (SSR) and cytochrome P450 gene based markers. A total of 18 RAPD primers, 10 SSR primers, and 10 pairs of cytochrome P450 gene based markers, respectively, revealed 49.4%, 50.2% and 58.7% polymorphism in 52 genotypes of E. coracana. Mean polymorphic information content (PIC) for each of these marker systems (0.351 for RAPD, 0.505 for SSR and 0.406 for cyt P450 gene based markers) suggested that all the marker systems were effective in determining polymorphisms. Pair-wise similarity index values ranged from 0.011 to 0.999 (RAPD), 0.010 to 0.999 (SSR) and 0.001 to 0.998 (cyt P450 gene based markers) and mean similarity index value of 0.505, 0.504 and 0.499, respectively. The dendrogram developed by RAPD, SSR and cytochrome P450 gene based primers analyses revealed that the genotypes are grouped in different clusters according to high calcium (300–450 mg/100 g), medium calcium (200–300 mg/100 g) and low calcium (100–200 mg/100 g). Mantel test employed for detection of goodness of fit established cophenetic correlation values above 0.95 for all the three marker systems. The dendrograms and principal coordinate analysis (PCA) plots derived from the binary data matrices of the three marker systems are highly concordant. High bootstrap values were obtained at major nodes of phenograms through WINBOOT software. Comparison of RAPD, SSR and cytochrome P450 gene based markers, in terms of the quality of data output, indicated that SSRs and cyt P450 gene based markers are particularly promising for the analysis of plant genome diversity. The genotypes of finger millet collected from different districts of Uttarakhand constitute a wide genetic base and clustered according to calcium contents. The identified genotypes could be used in breeding programmes and
Gómez, E.; E. J. Gómez; Matilde Padilla; González, A; Irene Expósito
El objetivo de este trabajo fue determinar el grado de utilización del nitrógeno, en dos cosechas de maíz rotadas con caupí (Vigna unguiculata Walp), debido al efecto en el suelo de varios tipos de abonos orgánicos. Para ello, se aplicaron al suelo cinco tipos de abonos orgánicos, de manera que cada uno de ellos aportara 475 kg.ha-1 de N total y se comparó la eficiencia en la utilización del N con un testigo sin abono. Los tratamientos se distribuyeron en el campo en Bloques al Azar. Se encon...
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Study on the safety and efficacy of miltefosine for the treatment of children and adolescents with post-kala-azar dermal leishmaniasis in Bangladesh, and an association of serum vitamin E and exposure to arsenic with post-kala-azar dermal leishmaniasis: an open clinical trial and case–control study protocol
Mondal, D; Hasnain, M G; Hossain, M S; Ghosh, D; Ghosh, P; Hossain, H; Baker, J; Nath, R; Haque, R; Matlashewski, G; Hamano, S
Introduction Post-kala-azar dermal leishmaniasis (PKDL) is a dermatological complication that occurs primarily among treated visceral leishmaniasis (VL) patients, and sporadically in a few without a history of VL. It mostly affects children and adolescents but is also common in adults. The conventional treatment with 120 intramuscular injections of sodium stibogluconate (SSG) is phasing out. Miltefosine (MF) is the only eventual alternative to SSG; however, its efficacy and safety profiles for treatment of children and adolescents with PKDL are lacking. In addition, risk factors for PKDL are poorly investigated. Host genetic, nutritional and environmental factors could be potential risk factors. As such, here we propose to evaluate the efficacy and safety of MF for 12 weeks at an allometric dose for children and adolescents with PKDL, and also to explore potential risk factors for PKDL. Methods and analysis A cross-sectional survey will look for suspected participants with PKDL among treated VL children and adolescents, a subsequent open clinical trial with MF at allometric dose, with a follow-up at 12 months. A case–control study will be carried out for PKDL risk factors. Assuming 95% cure rate, 95% CI and α=0.05, a sample size of 73 children with PKDL is needed. Considering an attrition rate of 10%, the final sample size is 80 children in each group. Descriptive and analytical analyses will be performed. Primary outcome is safety and cure rate of 12 weeks of treatment with MF. Ethics and dissemination International Centre for Diarrhoeal Disease Research, Bangladesh (ICDDR,B) Ethical Review Committee (ERC) approved the protocol (PR#013045). Written informed consent will be taken from all participants and their guardians (in case of minor). A Data and Safety Monitoring Board (DSMB) of ICDDR,B ERC will monitor all study activities to ensure the safety of the participants. Trial registration number NCT02193022; Pre-results. PMID:27188804
Cunha, Joana T; Ribeiro, Tânia I B; Rocha, João B; Nunes, João; Teixeira, José A; Domingues, Lucília
Serra da Estrela Protected Designation of Origin (PDO) cheese is the most famous Portuguese cheese and has a high commercial value. However, the adulteration of production with cheaper/lower-quality milks from non-autochthones ovine breeds compromises the quality of the final product and undervalues the original PDO cheese. A Randomly Amplified Polymorphic DNA (RAPD) method was developed for efficient detection of adulterant breeds in milk mixtures used for fraudulent production of this cheese. Furthermore, Sequence Characterized Amplified Region (SCAR) markers were designed envisioning the detection of milk adulteration in processed dairy foods. The RAPD-SCAR technique is here described, for the first time, to be potentially useful for detection of milk origin in dairy products. In this sense, our findings will play an important role on the valorization of Serra da Estrela cheese, as well as on other high-quality dairy products prone to adulteration, contributing to the further development of the dairy industry. PMID:27283677
Manoj P. Brahmane
Full Text Available We used random amplification of polymorphic DNA (RAPD to generate species-specific diagnostic fragment patterns for the molecular identification of the ornamental aquarium fish species Badis badis and Dario dario. Seven arbitrary oligodecamer primers produced a total of 116 bands of which 98.23% were polymorphic. The size of the amplified products was in the range 340 bp to 2170 bp. Intraspecies genetic similarity was 0.879 ± 0.023 for B. badis and 0.840 ± 0.014 for D. dario while interspecies genetic similarity was 0.602 ± 0.017, with cluster analysis displaying separate taxonomic and evolutionary status for these fish. The results show that RAPD was useful for the molecular identification of aquarium fish species, with morphological traits also being important.
Alexandre Wagner Silva Hilsdorf
Full Text Available Random amplified polymorphic DNA (RAPD markers were used to investigate the genetic relationship among nests of the carpenter ant, Camponotus rufipes, located in the same area. Five random oligodecamers were used to amplify DNA from 108 ant workers collected from six nests. A total of 47 RAPD markers were identified, which revealed low levels of genetic differentiation among nests (Fst = 0.00218 and a low average Shannon index (0.3727 among workers within nests. These results together suggest that the C. rufipes nest may be formed by a single, once-mated queen and that nests produced by queens that are genetically related tend to keep their nests in close proximity to one other.
Analysis of the genetic diversity in Metopolophium dirhodum (Walker (Hemiptera, Aphididae by RAPD markers Análise da diversidade genética de Metopolophium dirhodum (Walker (Hemiptera, Aphididae por meio de marcadores RAPD
Full Text Available The emergence of host-races within aphids may constitute an obstacle to pest management by means of plant resistance. There are examples of host-races within cereals aphids, but their occurrence in Rose Grain Aphid, Metopolophium dirhodum (Walker, 1849, has not been reported yet. In this work, RAPD markers were used to assess effects of the hosts and geographic distance on the genetic diversity of M. dirhodum lineages. Twenty-three clones were collected on oats and wheat in twelve localitites of southern Brazil. From twenty-seven primers tested, only four primers showed polymorphisms. Fourteen different genotypes were revealed by cluster analysis. Five genotypes were collected only on wheat; seven only on oats and two were collected in both hosts. Genetic and geographical distances among all clonal lineages were not correlated. Analysis of molecular variance showed that some molecular markers are not randomly distributed among clonal lineages collected on oats and on wheat. These results suggest the existence of host-races within M. dirhodum, which should be further investigated using a combination of ecological and genetic data.A emergência de raças hospedeiro-especialistas em afídeos pode constituir um obstáculo ao manejo de pragas por meio de plantas resistentes. Existem exemplos de raças hospedeiro-especialistas em afídeos de cereais, embora a ocorrência de raça hospedeiro-especialista no pulgão-verde-pálido-do-trigo Metopolophium dirhodum (Walker, 1849 (Hemiptera, Aphididae não tenha sido relatada ainda. Marcadores RAPD foram utilizados para avaliar os efeitos da distância geográfica e do hospedeiro sobre a diversidade genética de linhas clonais de M. dirhodum. Vinte e três clones foram coletados em aveia e trigo em doze localidades do sul do Brasil. De vinte e sete iniciadores usados para a análise, apenas quatro iniciadores mostraram polimorfismos. A análise de agrupamento por similaridade genética revelou haver quatorze
Divergência genética em tomate estimada por marcadores RAPD em comparação com descritores multicategóricos Genetic divergence among tomato accessions using RAPD markers and its comparison with multicategoric descriptors
Leandro SA Gonçalves
Full Text Available A estimativa da variabilidade genética existente em um banco de germoplasma é importante não só para a conservação dos recursos genéticos, mas também para aplicações no melhoramento de plantas. O presente trabalho teve como objetivo estudar a divergência genética entre 78 acessos de uma coleção de germoplasma de tomateiro, com base em 74 marcadores RAPD e correlacionar esses resultados àqueles da caracterização morfoagronômica realizada para 27 descritores. Foi utilizado o agrupamento hierárquico UPGMA para analisar os dados, observando-se a formação de 13 grupos. Esses grupos foram correlacionados a cinco descritores (hábito de crescimento, tipo de folha, cor do fruto, número de lóculos e formato do fruto. Alguns grupos apresentaram peculiaridades, a exemplo do grupo IV, que reuniu acessos com frutos no formato de pêra; o grupo VII com acessos resistentes a murcha-bacteriana e o grupo IX, que englobou acessos com folhas do tipo batata. As análises por bootstrap revelaram poucos agrupamentos consistentes. Houve correlação positiva e altamente significativa entre as matrizes geradas pelos 27 descritores qualitativos e pelos marcadores RAPD (t = 14,02. A correlação de Mantel (r = 0,39 foi altamente significativa, porém de baixa magnitude. O baixo valor verificado para esta correlação sugere que ambas as etapas de caracterização (morfoagronômica e molecular são importantes para um conhecimento mais amplo e melhor discriminação entre os acessos de tomate.The estimation of genetic variability in a germplasm bank is important not only for the conservation of the genetic resources, but also for applications in plant breeding. The genetic divergence among 78 tomato accessions was studied, based on 74 RAPD markers. Also, a correlation between the molecular profile and 27 morphological and agronomic data was performed. Cluster analysis (UPGMA, used to study the data, resulted in 13 groups that were correlated with
Eduardo Alano Vieira
Full Text Available Annual ryegrass is a temperate climate annual foraging grass, grown mostly in the South of Brazil, especially in the state of Rio Grande do Sul. Despite its importance, little is known about its genetic diversity, both within and among the populations cultivated. This knowledge is of fundamental importance for developing breeding and conservation strategies. The aim of this study was to characterize the genetic diversity and structure of four populations of annual ryegrass. Three of the populations were located in Rio Grande do Sul and the fourth in Uruguay. RAPD markers were used to study the genetic diversity and structure of these populations. Analysis of 375 individuals sampled from the populations, using six RAPD primers, generated a total of 82 amplified bands. They included 73 polymorphic bands (89,02%. The value of the total genetic diversity index obtained, (0,71 was high, indicating the presence of wide genetic diversity in the four populations. Genetic structure analysis revealed that 98% of total diversity is intrapopulational, whereas interpopulational genetic diversity was only 2%. These results suggest that before these populations separated, they had gone through a period of gene exchange and, even after the separation event, gene frequency stayed at levels similar to the original levels, with no differential selection for these genes in the different cultivation areas.O azevém anual é a gramínea anual forrageira de clima temperado de maior utilização no sul do Brasil, particularmente no Estado do Rio Grande do Sul. Apesar de toda a importância que a espécie apresenta, pouco se conhece a respeito da diversidade genética presente entre e dentro das populações cultivadas. Este conhecimento é de fundamental importância para o estabelecimento das estratégias de melhoramento genético e de conservação destes materiais. O objetivo deste estudo foi caracterizar a diversidade genética e a estrutura genética de quatro
Huh, Man; Lee, Hak; Lee, Bok; Choi, Joo
RAPD analysis was conducted to estimate genetic diversity and population structure of the wild (natural) and cultivated sea lettuce, Enteromorpha prolifera. The objectives of this study were to estimate the levels of genetic diversity in the wild and cultivated populations and to describe how the genetic variation of this species is distributed within and among its populations. In wild sea lettuce, 93.2% of loci at the species level showed polymorphism. The cultivated populations were found t...
Cunha, Joana T.; Ribeiro, Tânia I. B.; Rocha, João B.; Nunes, João; Teixeira, J. A.; Domingues, Lucília
Available online 17 May 2016 Serra da Estrela Protected Designation of Origin (PDO) cheese is the most famous Portuguese cheese and has a high commercial value. However, the adulteration of production with cheaper/lower-quality milks from non-autochthones ovine breeds compromises the quality of the final product and undervalues the original PDO cheese. A Randomly Amplified Polymorphic DNA (RAPD) method was developed for efficient detection of adulterant breeds in milk mixtures used for fra...
Maurizio Guida; Marco Guida; Bruna De Felice; Daniela Santafede; Raffaella D'Alessandro; Attilio Di Spiezio Sardo; Marianna Scognamiglio; Cinzia Ferrara; Giuseppe Bifulco; Carmine Nappi
The aim of this study was to assess the genotoxic effects of environmental chemicals on residents living near landfills. The study was based on samples of amniotic fluid from women living in the intensely polluted areas around the Campania region of Italy compared to a nonexposed control group. We evaluated the genetic effects that this amniotic fluids collected in contaminated sites had on Paracentrotus lividus embryos. DNA damage was detected through changes in RAPD (Random Amplified Polymo...
Full Text Available Os fungos Acremonium strictum e Fusarium verticillioides normalmente apresentam algumas similaridades morfológicas, o que dificulta sua diferenciação em sementes de milho (Zea mays, particularmente quando ocorrem simultaneamente. Técnicas moleculares de análise do DNA têm possibilitado o desenvolvimento de métodos rápidos, sensíveis eespecíficos no diagnóstico de fitopatógenos, em complemento à análise morfológica. Este trabalho objetivou caracterizar e dimensionar a diversidade genética de dez isolados de A. strictum obtidos de sementes de milho, provenientes de diferentes regiões produtoras brasileiras, por meio da análise do DNA genômico (RAPD. Objetivou-se ainda, diferenciar os isolados de A. strictum de F. verticillioides por meio da técnica citada. Pela análise do DNA, 25 primers Operon geraram polimorfismos, os quais tornaram possível e seguro o agrupamento de isolados de A. strictum e sua diferenciação de F. verticillioides. Os isolados de A. strictum apresentaram variabilidade intraespecífica entre 3,4% e 44,4%. Para a maioria dos casos não foi possível correlacionar a similaridade genotípica e a origem geográfica dos isolados de A. strictum.Differentiation between Acremonium strictum and Fusarium verticillioides infecting corn (Zea mays seeds is difficult because they present some morphological similarities. Molecular techniques have been applied with success, allowing the development of fast, sensible and specific diagnostic methods for plant pathogens in complement to morphological analysis. The objectives of this work were to characterize and measure the genetic diversity of ten isolates of A. strictum associated with maize (Zea mays seeds from different brazilian regions, using DNA analysis (RAPD in comparison with one isolate of F. verticillioides. Twenty-five Operon primers generated polymorphisms that were efficient in grouping A. strictum isolates and in distinguishing them from F. verticillioides
Roberto Pedroso de Oliveira
Full Text Available The objective of this work was to construct linkage maps of 'Pêra' sweet orange [Citrus sinensis (L. Osbeck] and 'Cravo' mandarin (Citrus reticulata Blanco using RAPD markers and the pseudo-testcross strategy. The parents were chosen according to the resistance/susceptibility to citrus variegate chlorosis (CVC. The segregation of 176 markers was analyzed in 94 progeny of F1 hybrids, which were obtained from controlled crossings. The linkage map of 'Pêra' sweet orange had 117 markers defined by 12 linkage groups, which spanned 612.1 cM. Only six markers could not be linked to the linkage group and 48.7% of the markers showed segregation distortion. The linkage map of 'Cravo' mandarin had 51 markers defined by 12 linkage groups, which spanned 353.3 cM. Only two markers did not link to the groups and 15.7% showed segregation distortion. The construction of linkage maps is relevant to future mapping studies of the inheritance of CVC, citrus canker and leprosis.O objetivo deste trabalho foi elaborar mapas de ligação de laranja 'Pêra' [Citrus sinensis (L. Osbeck] e tangerina 'Cravo' (Citrus reticulata Blanco utilizando marcadores RAPD e a estratégia pseudo-testcross. Os parentais foram escolhidos tomando-se por base o nível de resistência/suscetibilidade à clorose variegada dos citros (CVC. A segregação de 176 marcadores foi analisada em uma progênie de 94 híbridos F1, obtidos por meio de cruzamentos controlados. O mapa de ligação de laranja 'Pêra' apresentou 117 marcadores distribuídos em 12 grupos de ligação, totalizando 612,1 cM. Apenas seis marcadores não se ligaram a algum dos grupos de ligação e 48,7% deles apresentaram segregação distorcida. O mapa de ligação de tangerina 'Cravo' apresentou 51 marcadores distribuídos em 12 grupos de ligação, totalizando 353,3 cM. Apenas dois marcadores apresentaram-se desligados e 15,7% deles apresentaram segregação distorcida. A elaboração de mapas de ligação dessas
MENG Zining; ZHUANG Zhimeng; JIN Xianshi; TANG Qisheng; SU Yongquan
Random amplified polymorphic DNA (RAPD) technique is applied to 12 individuals from each species of the hairtail fishes Trichiurus lepturus and Eupleurogrammus muticus in the Yellow Sea. The percentage of polymorphic sites, degree of genetic polymorphism and genetic distance are compared and the phylogenetic tree is constructed by Neighbor-joining method. The partial mitochondrial 16S rRNA gene is amplified by polymerase chain reaction (PCR) and the PCR products are directly sequenced after being purified. These sequences, together with the homologous sequences of another Trichiuridae species Lepidopus caudatus obtained from GenBank, are used to analyze nucleotide difference and to construct a UPGMA phylogenetic tree by means of biological informatics. Analysis shows: (1) the RAPD technique is a highly sensitive method for investigating genetic diversity in T. lepturus, and E. muticus. T. lepturus exhibits a lower polymorphism and genetic diversity than E. muticus; (2) according to the analysis of the partial mitochondrial 16S rRNA gene sequences, a very low intraspecific variation and considerably high divergence among species were found, which reveals a dual nature of conservatism and variability in mitochondrial 16S rRNA gene; (3) five primers generate the species-specific RAPD sites and these sites can be served as the molecular markers for species identification and (4) it can be proved at DNA variation level that T. lepturus and E. muticus are of two species respectively pertaining to different genera, which supports the Nelson taxonomic conclusion.
Skibsted, U.; Baggesen, Dorte Lau; Dessau, R.; Lisby, G.
Isolates of Salmonella Enteritidis from 81 patients from Herlev Hospital or from Copenhagen County were analysed by random amplification of polymorphic DNA (RAPD), pulsed-field gel electrophoresis (PFGE) and phage-typing. Fourteen polymorphic markers from five decamer primers unambiguously placed...... that RAPD is useful as a tool in investigations of microbial outbreaks in its own right, or to supplement phage-typing and PFGE of Salmonella Enteritidis.......Isolates of Salmonella Enteritidis from 81 patients from Herlev Hospital or from Copenhagen County were analysed by random amplification of polymorphic DNA (RAPD), pulsed-field gel electrophoresis (PFGE) and phage-typing. Fourteen polymorphic markers from five decamer primers unambiguously placed...... (both PFGE type I) formed the fifth group. The sixth group of four isolates was not phage typeable and was PFGE type III. Forty outbreak-related isolates of phage-type 6 were resolved into three strains. No diversity of phage-type 6 was found among isolates unrelated to the outbreak. It is concluded...
LI Xiaozun; PAN Junsong; WANG Gang; TIAN Libo; SI Longting; WU Aizhong; CAI Run
A cucumber ( Cucumis sativus L. ) molecular linkage map, including 79 random-amplified polymorphic DNAs (RAPD)and two genes , lb for lateral branch and f for female sex expression, is constructed from a cross between a line, S52, with weak lateral growing ability and staminate from Dabieshan Mountains area in China and another line, S06, with strong lateral growing ability and gynoecious from Europe. The map contains nine linkage groups and spans 1110.0 cM with an average distance of 13.7 cM between loci. The lb locus is located in a longer linkage group LG-2 and flanked by two markers, OP-Q5-1 and OP-M-2-2, at 9.3 cM and 15.9 cM, respectively. In the meantime, the RAPD loci, OP-Q5-2 and BC151, in a short linkage group were found to flank f at 13.7 cM and 13.4 cM,respectively. The construction of RAPD map has paved a way for further study of the genes for lateral branch, female sex expression and other agronomic traits in cucumber.
Tereza Cristina A. Leal
Full Text Available Sixteen strains of Yersinia enterocolitica serotype O:3, isolated from apparently healthy pigs collected in Rio de Janeiro, and four human strains of serotypes O:4, O:5, O:6 and O:13 were analyzed by RAPD-PCR. The strains were grouped into five genotypic profiles according to the amplification patterns obtained with three random primers. Fifteen of the 16 pig strains had identical amplification patterns, which was named genotypic profile 1. The one different profile was named genotypic profile 2. Genotypic profile 1 was also exhibited by the O:6 human serotype strain. The O:4 and O:13 human serotype strains showed similar amplification profiles with two primers. However, the third primer induced a distinct profile in each strain. Therefore, these two strains were placed into genotypic profile 3 and 4, respectively. Each primer produced a completely different amplification profile in the O:5 human serotype strain; therefore, it was named genotypic profile 5. The presence or absence of plasmids in the strains studied did not affect the amplification results. These results show that genetic variations can exist within a serotype, and strains of different serotypes can exhibit the same amplification profile when compared using other primers.Foram utilizados três "primers" aleatórios para caracterizar pela técnica RAPD-PCR 16 cepas de Yersinia enterocolitica do sorotipo O:3, isoladas de suínos sadios do Rio de Janeiro. Pelos resultados dos padrões de amplificação, as 16 cepas dos suínos e as 4 cepas humanas usadas como referência (sorotipos O:4, O:5, O:6 e O:13 foram agrupadas em 5 perfis genotípicos. Quinze cepas de suínos apresentaram um padrão de amplificação idêntico (perfil genotípico 1 e somente uma apresentou um perfil de amplificação diferente (perfil genotípico 2. O mesmo padrão de amplificação do perfil genotípico 1 foi também observado em uma cepa humana do sorotipo O:6. As cepas humanas dos sorotipos O:4 e O:13
Zhou, L; Wang, Y; Gui, J F
Sex evolution has been a debating focus in evolutionary genetics. In lower vertebrates of reptiles, amphibians, and fish, a species or a bioform reproduces either sexually or asexually but never both. A few species were found to consist of all females in fish. These all-female species can propagate by asexual reproduction modes, such as gynogenesis and hybridogenesis. However, the coexistence of sexuality and asexuality in a single species was recently noted only in a cyprinid fish silver crucian carp, Carassius auratus gibelio. This fish had been demonstrated to be capable of gynogenesis stimulated by sperm from other related species. Surprisingly, natural populations of this fish consist of a minor but significant portion (approx. 20%) of males. As different clones with specific phenotypic and genetic characteristics have been found, and RAPD markers specific to each clone have recently been identified, this fish offers many advantages for analyzing whether or not genetic recombination occurs between different clones. In this study, artificial propagation was performed in clone F and clone D. Ovulated eggs from clone F were divided into two parts and respectively inseminated with sperm from a clone D male and from a red common carp (Cyprinus carpio) male. The control clone D individuals were selected from gynogenetic offspring of clone D activated by sperm of red common carp. The phenotype and sex ratio in the experimental groups were also observed. Using RAPD molecular markers, which allow for reliable discrimination and genetic analysis of different clones, we have revealed direct molecular evidence for gonochoristic reproduction in the gynogenetic silver crucian carp and confirmed a previous hypothesis that the silver crucian carp might reproduce both gynogenetically and gonochoristically. Therefore, we conclude that the silver crucian carp possesses two reproductive modes, i.e., gynogenetic and gonochoristic reproduction. The response mechanism of two
Ana Lilia Alzate-Marin
Full Text Available An efficient and rapid DNA minipreparation modified method for frozen samples was developed for five tropical tree species: Copaifera langsdorffii, Hymenaea courbaril, Eugenia uniflora, Tabebuia roseo alba and Cariniana estrellensis. This procedure that dispenses the use of liquid nitrogen, phenol and the addition of proteinase K, is an adaptation of the CTAB-based DNA extraction method. The modifications included the use of PVP to eliminate the polyphenols, only one chloroform-isoamyl alcohol step and the addition of RNase immediately after extraction with chloroform. The yields of the DNA samples ranged from 25.7 to 42.1 µg from 100 mg leaf tissue. The DNA samples extracted by this method were successfully used for PCR (SSR and RAPD analyses in these five and other twelve tropical tree species.Este trabalho teve como objetivo otimizar um protocolo econômico, rápido e eficaz de minipreparação de DNA genômico, para as espécies florestais Copaifera langsdorffii (Óleo de Copaíba, Hymenaea courbaril (Jatobá, Eugenia uniflora (Pitanga, Tabebuia roseo alba (Ipê Branco e Cariniana estrellensis (Jequitibá Branco. Este método é uma adaptação da técnica de extração CTAB de Doyle e Doyle (1990, o qual consiste principalmente na adição de PVP para eliminar polifenoles, somente uma etapa de extração com clorofórmio-álcool isoamílico e a adição da RNase A imediatamente após a extração com clorofórmio. O método também dispensa o uso de nitrogênio líquido, o uso do fenol e a adição de proteinase K. Os DNAs das espécies florestais extraídos apresentaram alto rendimento e boa qualidade, com rendimento de 25.7 a 42.1 µg de DNA a partir de 100 mg de tecido foliar congelado. Com este protocolo, em apenas 1 dia de trabalho, uma pessoa pode completar o isolamento do DNA de aproximadamente 50 amostras de folhas (dependendo da capacidade da centrífuga. O DNA obtido pode ser usado para métodos de análise baseados em PCR (SSR e
蒋建平; 杨懋勋; 黄永芳
本研究选择了中国广东省7个有代表性的山区作为野百合(Lilium brownii F.E.Brown ex Miellez)天然居群的采样点,对采集到的199份样品进行了随机扩增多态性DNA(RAPD)分析,以建立野百合天然居群RAPD分析方法体系以便后续研究,并了解广东省野百合天然居群多态性情况和居群内外个体及野百合及变种百合样品的聚类情况.以新鲜叶片、硅胶干燥叶片及鳞片为材料分别提取DNA,从273条10聚寡核苷酸随机引物中筛选出20条随机引物,对7个居群共199个野百合及百合样品进行RAPD扩增.共检测到433条RAPD谱带,其中多态性条带为430条,所有个体的多态性条带百分率(PPB)为99.31％,各居群的平均多态位点比率为60.67％.通过对199个个体的RAPD数据的UPGMA聚类分析,获得了聚类图.199个样品用三种方法都成功提取出质量较高的DNA,并能扩增出清晰、明亮、稳定的RAPD条带.在聚类图遗传相似系数为0.68处发现同一居群内的所有百合个体和野百合个体在聚类图中表现了明显的分离,这是原变种野百合及变种百合的区别在分子水平上很好的反映.%Lilium brownii F. E. Brown ex Miellez, Brown lily, belongs to Lilium of Liliaeeae in planttax-onorny, is a endemic species and one of commonlly-used cross breeding parents of Lilium in China. L. Brownii F. E. Brown ex Miellez var. Virdulum Baker, Greenish lily, a variety of Lilium brownii, is also a well known medicinal and edible plant. Wild germplasm resources are the basic materials for genetic research and plant breeding. Howerever, due to Lilium brownii is scattered in the wild and its collection is generally only in flowering phase, it is a great difficulty for its scientific research. In the year before, researches on Lilium brownii especially molecular biology researches are still lacking in Guangdong province. In this study, total 199 Lilium brownii samples of 7 natural populations collected from
野蘑菇(Agaricus arvensis Schaeff ex.Fr.)是蘑菇属(Agaricus)又一种近年来广泛栽培的食用菌.由于它的菌丝细胞具有多核,无锁状联合,以及人们对它的繁殖模式和生活史认识的不足,给杂交育种工作造成了较多困难.运用随机扩增多态DNA遗传标记技术,结合拮抗试验和核相分析,对自育的单孢菌株之间的杂交试验进行分析研究.结果表明:两个相互亲合的同核体菌株被配对培养时,交配反应出现,并形成异核体的后代.可能野蘑菇具有双重的交配繁殖系统--同宗配合和异宗配合.在食用菌杂交育种研究中,机扩增多态DNA(RAPD)是一种非常有效和方便的检验杂合子的方法.
Full Text Available CONTEXTO: A literatura mostra comorbidade entre jogo patológico, abuso de substâncias e transtornos afetivos (maior prevalência de depressão em adultos. É alta a prevalência de depressão nos idosos, tornando-os vulneráveis à dependência de jogos de azar e ao abuso de substâncias. Com o aumento desta população em todo o mundo, faz-se necessário mais pesquisas sobre esse tema. OBJETIVOS: Avaliar o grau de envolvimento do idoso que aposta em jogos de azar e sua correlação com uso de substâncias. MÉTODOS: Revisão bibliográfica dos bancos de dados MEDLINE, LILACS e PsycINFO, sem limite de tempo. RESULTADOS: Idosos apostadores recreativos de jogos de azar apresentam consumo principalmente de álcool e boa avaliação de saúde geral. Idosos com problemas com jogo apresentam abuso de álcool, mais transtornos psiquiátricos e doenças cardiovasculares que os idosos jogadores recreativos. Quando comparados com adultos ou jovens jogadores, os idosos apresentam menos problemas relacionados ao consumo de álcool. CONCLUSÕES: Assim como nos adultos, também existe correlação entre apostas em jogos de azar e consumo de substâncias nos idosos. Quanto mais problemático o envolvimento com jogo, mais problemático é o consumo de substâncias (álcool e maior prevalência de transtornos do humor (depressão. No entanto, não se pode afirmar uma relação causal entre esses fatores, sendo necessária realização de mais pesquisas sobre esse tema.BACKGROUND: Literature presents comorbidity between pathological gambling, substance use/abuse and affective disorders (depression being the most prevalent in adults. The prevalence of depression disorder in elderly people is high, making them vulnerable to pathological gambling and substance abuse. With the increase of this population worldwide, more research regarding these issues has become necessary. OBJECTIVES: Assess the involvement of the elderly in gambling activities and its correlation
Liu, Bo; Wang, Qingyin; Li, Jian; Liu, Ping; He, Yuying
The Chinese shrimp Penaeus ( Fenneropaeneus) chinensis is an important species in marine fishery and aquaculture in China. A female Chinese shrimp Penaeus ( Fenneropaeneus) chinensis was captured from west coast of the Korean peninsula and mated with a “Yellow Sea No. 1” male to produce the first filial generation (F1) 100 F2 full-sib progeny from brother-sister crosses between F1 families was used for the mapping study. A genetic linkage map of the Chinese shrimp was constructed, based on 354 markers, including 300 amplified fragment length polymorphism (AFLP) markers, 42 microsatellite (SSR) markers, and 12 randomly amplified polymorphism (RAPD) markers. Forty-seven linkage groups (LGs) were identified. The total map length was 4 580.5 cM, with an average spacing of 11.3 cM, covering 75.8% of the estimated genome size. The construction of this genetic linkage map was part of a genetic breeding program. This linkage map will contribute to the discovery of genes and quantitative trait loci (QTLs) in Chinese shrimp.
Full Text Available Plant tissue culture techniques offer a great opportunity to overcome the limitations associated with the large scale cultivation of spine gourd. Present study was carried out to formulate the best possible media for large scale production of spine gourd and result of the study revealed that highest percentage (85% of embryogenic callus was obtained from MS medium supplemented with 2.0 mg/L each of 2, 4- Dichlorophenoxyacetic acid (2, 4-D and 6- Benzylamino Purine (BAP in leaf explants of spine gourd. Maximum number of shoots (12.15 ± 1.51 shoots were observed on MS medium augmented with BAP (4.0 mg/L in combination with L-glutamine (2.0 mg/L from leaf derived embryogenic callus of spine gourd. Identification of sex by using morphological characters in the newly regenerated plantlets of spine gourd at fourth leaf stage is another problem for large scale propagation of female plants. PCR based molecular marker OPA-15, a Random Amplified Polymorphic DNA (RAPD primer can be used as a differential marker to identify female plants form male plants at pre-flowering stage in newly regenerated plantlets (in vitro and as well as in field plants (in vivo of spine gourd. A unique amplification band (700 bp in size appeared only in female samples, but not in male samples of spine gourd.
Saha, Soumen; Adhikari, Sinchan; Dey, Tulsi; Ghosh, Parthadeb
Plant regeneration through rapid in vitro clonal propagation of nodal explants of Morus alba L. variety S-1 was established along with genetic stability analysis of regenerates. Axillary shoot bud proliferation was achieved on Murashige and Skoog (MS) medium in various culture regimes. Highest number of shoots (5.62 ± 0.01), with average length 4.19 ± 0.01 cm, was initially achieved with medium containing 0.5 mg/l N(6)-benzyladenine (BA) and 3% sucrose. Repeated subculturing of newly formed nodal parts after each harvest up to sixth passage, yielded highest number of shoots (about 32.27) per explants was obtained after fourth passage. Rooting of shoots occurred on 1/2 MS medium supplemented with 1.0 mg/1 Indole-3-butyric acid (IBA). About 90% (89.16) of the plantlets transferred to the mixture of sand:soil:organic manure (2:2:1) in small plastic pots acclimatized successfully. Genetic stability of the discussed protocol was confirmed by two DNA-based fingerprinting techniques i.e. RAPD (random amplified polymorphic DNA) and ISSR (inter-simple sequence repeat). This protocol can be used for commercial propagation and for future genetic improvement studies. PMID:26693403
Young shoots of Dahlia pinnata Cav cultivar were irradiated by 80 MeV/u 12C6+ ions. The differences at DNA level between dwarf mutant and wild type were detected by using RAPD (Random amplified polymorphic DNA) method. The results showed that there were different polymorphic bands induced by 18 primers among the tested 25 primers, and the ratio of polymorphic bands and total bands accounted for 19.57% after 1.80 x 108C6+/cm2 irradiation. Meanwhile, after 1.08 x l08 C6+/cm2 irradiation the polymorphic bands were induced by only 6 primers and the ratio accounted for 5.76%. Using the Jaccard formula, statistic analysis of amplified bands were obtained and it showed that after 1.80 x 108 C6+/cm2 and 1.08 x 108 C6+/cm2 irradiation the similarity index were 0.65 and 0.92 respectively. The results suggested that DNA should prone to mutant at high doses, and high dose irradiation might be more effective in corn improving and mutagenic breeding. (authors)
Lu, Pei-Luen; Yorkson, Mitsuko; Morden, Clifford W
The genus Chrysodracon has six endemic species in the Hawaii Islands. Chrysodracon hawaiiensis is endemic to Hawaii Island and was described as a distinct species in 1980. It was listed as an endangered species on the International Union for the Conservation of Nature and Natural Resources (IUCN) Red List in 1997. This woody plant species was, at one time, common in exposed dry forests, but it became very rare due to grazing pressure and human development. The tree species Chrysodracon auwahiensis (C. auwahiensis), endemic to Maui and Molokai, still has large adult populations in dry lands of the islands, but unfortunately no regeneration from seed has been reported in those areas for many years. The two endemic species were examined using the molecular technique of random amplified polymorphic DNA (RAPD) and inter simple sequence repeats (ISSR) to determine the genetic structure of the populations and the amount of variation. Both species possess similar genetic structure. Larger and smaller populations of both species contain similar levels of genetic diversity as determined by the number of polymorphic loci, estimated heterozygosity, and Shannon's index of genetic diversity. Although population diversity of Chrysodracon hawaiiensis (C. hawaiiensis) is thought to have remained near pre-disturbance levels, population size continues to decline as recruitment is either absent or does not keep pace with senescence of mature plants. Conservation recommendations for both species are suggested. PMID:27537876
Full Text Available The genus Chrysodracon has six endemic species in the Hawaii Islands. Chrysodracon hawaiiensis is endemic to Hawaii Island and was described as a distinct species in 1980. It was listed as an endangered species on the International Union for the Conservation of Nature and Natural Resources (IUCN Red List in 1997. This woody plant species was, at one time, common in exposed dry forests, but it became very rare due to grazing pressure and human development. The tree species Chrysodracon auwahiensis (C. auwahiensis, endemic to Maui and Molokai, still has large adult populations in dry lands of the islands, but unfortunately no regeneration from seed has been reported in those areas for many years. The two endemic species were examined using the molecular technique of random amplified polymorphic DNA (RAPD and inter simple sequence repeats (ISSR to determine the genetic structure of the populations and the amount of variation. Both species possess similar genetic structure. Larger and smaller populations of both species contain similar levels of genetic diversity as determined by the number of polymorphic loci, estimated heterozygosity, and Shannon’s index of genetic diversity. Although population diversity of Chrysodracon hawaiiensis (C. hawaiiensis is thought to have remained near pre-disturbance levels, population size continues to decline as recruitment is either absent or does not keep pace with senescence of mature plants. Conservation recommendations for both species are suggested.
谭冬梅; 罗淑萍; 李疆; 韩海涛
应用随机扩增多态性DNA(RAPD)技术对新疆阿月浑子(Pistacia vera L.)地方品种雌、雄株进行性别鉴定的研究.通过叶片总DNA抽提、RAPD标记分析及反复的试验筛选,找到了一个适用于新疆阿月浑子的RAPD反应体系和循环参数.采用OPO08引物对雌、雄株基因组DNA扩增出性别之间差异性的核苷酸片段,证实雄性植株DNA的扩增产物有一条大约700bp的特异性条带,而雌性植株则无此特异性条带.即找到一条与阿月浑子性别相关的基因标记,表明RAPD技术可应用于新疆阿月浑子地方品种的性别鉴定.此研究是对我国雌雄异株果树阿月浑子在分子水平进行早期性别鉴定的一个尝试.
The aim of our study was to induce with radiation of atrazine resistant and tolerated mutants in Coles, Amsoy-71 and 1937 soybean varieties. Atrazine that is photosynthetic inhibitor is the most important herbicide of S-triazin group, and shows toxic effect on soybean plant. For the improvement of the atrazine resistant plants with mutation breeding, the seeds belonging to the three varieties were irradiated with 200 Gy of gamma radiation dose. The irradiated seeds were sown in the field and at the end of harvesting season, every pod at node situated on the main stem was picked up separately and M2 generations were obtained. At the plants, which were obtained from M2 generation, chlorophyll mutants were determined and atrazine selection was made. The percentage of chlorophyll mutants for Amsoy-71, Coles and 1937 soybean varieties were found as 1.07, 1.48 and 1.32, respectively. At the end of atrazine selection, the percentages of atrazine resistant plants for Amsoy-71, Coles and 1937 soybean varieties were 0.80, 0.60 and 0.53, respectively. The percentages of atrazine tolerated plants were 1.07, 1.18 and 1.05, respectively as well. In our research; the differences among the mutants replying to atrazine in various concentrations were examined by using RAPD procedure as the molecular marker techniques in comparison with polymorphism. In the study done by using 14 primers; according to the amplification results, the differences between atrazine resistant plants were shown
Full Text Available Objective(sPseudomonas aeruginosa is the most important cause of chronic lung infections and death in patients with cystic fibrosis. Determining the distribution of specific strains within patient populations is important in order to examine the epidemiology of the disease and the possibility of cross infection among patients. Materials and MethodsForty six Iranian patients with cystic fibrosis were studied for colonization with P. aeruginosa. Colony phenotype was recorded and antibiotic susceptibility to 11 antibiotics was determined using the disc diffusion method. Genetic fingerprinting was carried out by RAPD–PCR and by PFGE.ResultsForty five P. aeruginosa isolates were recovered from 31 patients including sequential cultures from 9 subjects. The rate of colonization increased with age. All isolates were susceptible to tobramycin and ciprofloxacin, 97.8% were sensitive to amikacin and piperacillin, 93.3% to gentamycin, 91.1% to ticarcillin, 86.7% to colistin, 80% to carbenicillin, 48.9% to cefotaxime, 26.7% to imipenem and 11.1% to ceftazidime. Genetic fingerprinting showed similar distribution profiles for RAPD-PCR and PFGE and the majority of the isolates had unique fingerprints. ConclusionNo relationship was observed between the obtained genotypes and antibiotic susceptibility profiles and common predominant virulent clones were not found among the isolates.
Coelho-Bortolo, T; Mangolin, C A; Lapenta, A S
Lasioderma serricorne (F.) is a small cosmopolitan beetle regarded as a destructive pest of several stored products such as grains, flour, spices, dried fruit and tobacco. Chemical insecticides are one of the measures used against the pest. However, intensive insecticide use has resulted in the appearance of resistant insect populations. Therefore, for the elaboration of more effective control programs, it is necessary to know the biological aspects of L. serricorne. Among these aspects, the genetic variability knowledge is very important and may help in the development of new control methods. The objective of this study was to evaluate the genetic variability of 11 natural populations of L. serricorne collected respectively in three and four towns in the states of Paraná and São Paulo, Brazil, using 20 primers random amplified polymorphic DNA (RAPD) and polymorphisms of esterases. These primers produced 352 polymorphic bands. Electrophoretic analysis of esterases allowed the identification of four polymorphic loci (Est-2, Est-4, Est-5 and Est-6) and 18 alleles. Results show that populations are genetically differentiated and there is a high level of genetic variability within populations. The high degree of genetic differentiation is not directly correlated to geographical distance. Thus, our data indicate that movement of infested commodities may contribute to the dissemination of L. serricorne, facilitating gene flow. PMID:26459013
Although bananas and plantains are one of the most important fruit crops, gearing up the breeding programmes for these has always remained the most difficult task due to several inherent problems such as parthenocarpy, barriers in obtaining viable seeds and long life cycle etc. In this regard, incorporation of in vitro techniques such as shoot-tip / cell cultures along with conventional as well as non-conventional methods of genetic improvement is of utmost importance, especially in those vegetatively propagated species with long crop cycle and low in vivo proliferation rate. In order to understand the radio-sensitivity, the callus and cell cultures of banana were exposed to differential doses of gamma-rays. Growth of the callus cultures reduced with increasing dose of gamma-rays. Similar trend was noticed in irradiation of cell suspensions also where a dose of 40 Gy and more was completely lethal. The experience gained from previous and present experiments has yielded optimization of the procedures for gamma-irradiation and subsequent handling of banana in vitro cultures. The RAPD analysis of the selected variants was unable to detect adequate polymorphism, and further experimentation in these regards is being done. (author)
Khrisanfova, G G; Ludannyĭ, R I; Slyn'ko, Iu V; Iakovlev, V N; Cemenova, S K
The polymerase chain reaction with arbitrary primers (RAPD-PCR) was used to study and to evaluate the genetic variation in the hybrid progeny of two Cyprinidae species, common bream Abramis brama and roach Rutilus rutilus. Genetic polymorphism was studied in 20 fishes (young of the current year) obtained in four individual crosses: R. rutilus x R. rutilus (RR), A. brama x A. brama (AA), R. rutilus x A. brama (RA), and A. brama x R. rutilus (AR). Amplification spectra obtained with eight primers contained 288 fragments, 97.6% of which proved to be polymorphic. The proportion of polymorphic fragments was 75.0% in the RR progeny, 58.1% in the AA progeny, 84.9% in the AR progeny, and 77.8% in the RA progeny. Classification analysis in the space of principal components was performed with the first four components, which together accounted for 64% of the total variance of the character under study. The individual contributions of components I, II, III, and IV were 26.8, 16.8, 11.5, and 8.9%, respectively. Fishes of the two pure species and the hybrid progeny (direct and reverse hybrids together) were clearly differentiated in the space of principal components I and II. The best differentiation of the four samples (RR, AA, RA, and AR) was observed in the space of principal components II and IV. Possible causes of high genetic variation in interspecific hybrids are discussed. PMID:15575514
Maria Sueli Papa Leuzzi
Full Text Available In this study, the RAPD technique was used to analyze the genetic structure of populations of the fish Astyanax altiparanae (Characidae, Tetragonopterinae living in the lower, middle and upper Paranapanema River, Brazil. The aim was to assess this structure regarding fish handling and conservation programs. The genetic variability (P was found to be 42.64%, 75% and 75% in the low, middle and upper reaches, respectively. The dendrogram of genetic similarity, obtained by comparative analysis of the sets of samples from the three sites, showed the formation of three clusters. All of the genetic parameters used indicate that the population in the lower Paranapanema is genetically different from those in the middle and upper sections. The theta P test shows that the low Paranapanema is highly differentiated from the middle (0.2813 and upper (0.2912 Paranapanema, while the differentiation between the last two is moderate (0.0895. The data obtained in the present work suggest that recolonization and conservation studies should not be focused on the species A. altiparanae as such, but on the conservation units, because they are the genetically differentiated populations.
Análisis de variabilidad genética en Moniliophthora roreri con AP-PCR y RAPD en Antioquia, Colombia Analysis of genetic variability in Moniliophthora roreri with AP-PCR and RAPD in Antioquia, Colombia
Afanador Kafuri Lucía
Full Text Available Moniliophthora roreri es el agente causante de la moniliasis del cacao, la enfermedad más severa en las plantaciones de cacao en el departamento de Antioquia, Colombia. Los marcadores moleculares RAPD (Random Amplyfied Polymorphism of DNA y AP-PCR (Arbitraly Primed Polymerase Chain Reaction fueron usados para estudiar la variabilidad genética de 170 aislamientos de M. roreri colectados en doce municipios de Antioquia. El análisis dividió la población en seis grupos, el grupo G1 fue el más grande y contenía el 95% de los aislamientos con una alta similitud genética (coeficiente de similitud de 0,7 a 1, mientras los otros cinco grupos contenían solo aislamientos de Apartadó y Dabeiba con una similitud genética moderadamente baja (coeficiente de similitud entre 0,45 a 0,55. El análisis de componentes principales mostró una alta similitud genética entre la población excepto entre los aislamientos de Apartadó y Dabeiba, que registraron los más altos niveles de variabilidad genética con valores altos del índice de Shannon y el porcentaje de loci polimórficos, mientras los otros aislamientos registraron una baja variabilidad genética. Los valores de diversidad y diferenciación genética en la población muestran una introducción reciente de M. roreri en las plantaciones de cacao de Antioquia, y una reproducción predominantemente clonal en la población. De acuerdo con Amova, la mayoría de la variación genética se encontró dentro de los municipios (75,68% con solo un 5,94% presente entre las subregiones.Moniliophthora roreri is the causal agent of moniliasis in cocoa, the most severe disease affecting cocoa plantations in the Antioquia department in Colombia. RAPD (random amplified polymorphism of DNA and AP-PCR (arbitrarily-primed polymerase chain reaction molecular markers were used for studying the genetic variability of 170 M. roreri isolates collected from twelve municipalities in Antioquia. Cluster analysis divided the
Rejane Rodrigues de Oliveira
Full Text Available The objective of this study was to verify the genetic diversity between and within seven populations of Moxotó goat (n = 264 from the States of Pernambuco, Paraíba and Rio Grande do Norte, using RAPD (Random Amplified Polymorphic DNA. Moxotó, as well as other naturalized breeds, suffers genetic losses due to the indiscriminate miscegenation with breeds raised in the Northeast Region of Brazil. The genetic characterization of these genetic resources is essential to conservation and breeding programs. DNA was extracted from lymphocytes using a non-organic protocol. The 16 primers used were selected from 120 decamer oligonucleotide primers and generated 56 polymorphic bands. The analysis of molecular variance (AMOVA showed that the greater part of total genetic variability (71.55% was due to differences between individuals within populations, while 21.21% was among populations. The analysis of variance among the pairs of populations demonstrated that the populations located in Floresta, PE x Angicos, RN presented a smaller value of intrapopulational differentiation (8.9%, indicating low genetic variability among them. Nei's genetic distances varied between 0.0546 and 0.1868 in the populations. The dendrogram generated showed that the Canindé breed, used as outgroup, clustered with the populations of Moxotó, indicating a possible common origin of the naturalized goat breeds.O objetivo deste estudo foi verificar a diversidade genética entre e dentro de sete populações de cabras Moxotó (n = 264 dos Estados de Pernambuco, Paraíba e Rio Grande do Norte, usando a técnica de RAPD (Poliformismo de DNA Amplificados ao Acaso. A raça Moxotó, assim como outras naturalizadas, sofre grande erosão genética devido à miscigenação indiscriminada com outras raças existentes na Região Nordeste. A caracterização genética desta raça é essencial para programas de conservação e melhoramento. Os marcadores moleculares são uma ferramenta útil na
Diversidade genética de Enterolobium contortisiliquum (Vell. Morong. no Baixo Rio São Francisco, por meio de marcadores RAPD Genetic diversity of Enterolobium contortisiliquum (Vell. Morong. in the low San Francisco river by RAPD markers
Georgea da Cruz Santana
Full Text Available Enterolobium contortisiliquum Vell. Morong (Leguminosae-Mimosoideae é uma espécie muito utilizada em programas de recuperação de matas ciliares no Baixo Rio São Francisco, devido ao seu rápido crescimento inicial. Assim, o objetivo deste trabalho foi avaliar, por meio de marcadores moleculares RAPD, a diversidade genética de oito indivíduos de uma população remanescente dessa espécie, visando contribuir para a definição de estratégias de coleta de sementes. Os indivíduos estão situados em uma área de 100 ha de mata ciliar do Baixo Rio São Francisco. Para a extração do DNA, pelo método CTAB 2%, foram utilizadas folhas tenras dos indivíduos. Testaram-se 20 oligonucleotídios de 10 bases de seqüência arbitrária, cujos produtos foram separados em gel de agarose 0,8%, submetidos à eletroforese horizontal, corados com brometo-de-etídio e visualizados em luz ultravioleta. A similaridade genética entre os indivíduos foi calculada pelo Coeficiente de Similaridade de Jaccard e a construção do dendrograma, realizada utilizando-se o método UPGMA. O valor médio de diversidade genética entre as matrizes foi de 49%, variando de 33 a 85%. Os indivíduos 6 e 7 apresentaram relativa proximidade genética (67%, não sendo indicado o plantio de suas mudas ou semeadura direta para recuperação de área ciliar em locais muito próximos. A partir dos resultados observados, podem-se desenvolver estratégias para a coleta de sementes e produção de mudas, auxiliando, assim, programas de restauração ambiental.Enterolobium contortisiliquum Vell. Morong (Leguminosae-Mimosoideae is very much used in riparian forest restoration programs in the Low San Francisco River because of its fast initial growth. The objective of this work was to evaluate by RAPD molecular markers the genetic diversity of eight individuals of a remaining population of this species, in order to contribute for the definition of strategies for seed production. The
Mapeamento de genes de resistência do feijoeiro à ferrugem, antracnose e mancha-angular usando marcadores RAPD Mapping rust, anthracnose and angular leaf spot resistance genes in common bean using RAPD markers
Fábio G. Faleiro
Full Text Available A organização de diferentes genes de resistência da cultivar Ouro Negro de feijoeiro-comum (Phaseolus vulgaris à ferrugem, antracnose e mancha-angular foi estudada com o auxílio de marcadores moleculares. Uma população de 154 linhas endogâmicas recombinantes (RIL's obtidas do cruzamento entre as cultivares Ouro Negro e Rudá foram inoculadas com sete raças fisiológicas de Uromyces appendiculatus, três de Colletotrichum lindemuthianum, e quatro de Phaeoisariopsis griseola. Amostras de DNA de cada uma das RIL's foram amplificadas via PCR utilizando 70 diferentes primers. A análise da segregação da resistência à ferrugem, antracnose e mancha-angular na população de 154 RIL's revelou diferentes modos de herança para a resistência a cada uma das raças fisiológicas. A análise de ligação genética revelou que os diferentes genes de resistência à ferrugem e à antracnose estão no mesmo grupo de ligação. Os genes de resistência à mancha-angular também foram mapeados juntos, porém em outro grupo de ligação. Verificou-se neste trabalho que a utilidade dos marcadores RAPD, previamente identificados como ligados a genes de resistência do feijoeiro a doenças foi restrita. Apenas cinco dos 38 marcadores moleculares testados foram validados na população de RIL's como ligados aos genes de resistência à ferrugem e à antracnose. Três novos marcadores (OBA16(669 e OBA16(583 a 10,4 cM em acoplamento e OAD9(3210 a 13,9 cM em repulsão ligados ao bloco gênico de resistência da cultivar Ouro Negro à mancha-angular foram identificados.Molecular markers were used to study the organization of rust, anthracnose and angular leaf spot resistance genes in common bean (Phaseolus vulgaris Ouro Negro cultivar. A segregant population of 154 recombinant inbred lines (RIL´s from the crossing between Ouro Negro and Rudá cultivars was inoculated under controlled conditions with seven races of Uromyces appendiculatus, three of
Cadena, Marleny; Naranjo, Alvaro; Ñústez, Carlos E.
Se evaluó el daño en tubérculo causado por la polilla guatemalteca (Tecia solanivora P.) en 60 genotipos de la especie Solanum phureja bajo condiciones de laboratorio en la Universidad Nacional de Colombia. Se utilizó un diseño completamente al azar con cinco repeticiones, tomando como unidad experimental un tubérculo, sobre el cual se colocaron 20 huevos de T. solanivora. Las variables evaluadas fueron: porcentaje de eclosión, porcentaje de pupamiento y porcentaje de daño. Los resultados obt...
Furong WANG; Zhijun ZHANG; Renzhong LIU; Qinhong LIU; Jun ZHANG
@@ Sixteen cultivars of Upland cotton (Gossypium hirsutum L. ) cultivars in Huang-Huai Cottongrowing Region were detected by RAPD while the F1 heterosis of each hybrid involved these cultivars were evaluated. The genetic similarity (GS) of the 16 cultivars through analysis of 115polymorphic RAPD loci obtained from 70informative primers were 53%～88%.
Assessment of genetic diversity on a sample of cocoa accessions resistant to witches' broom disease based on RAPD and pedigree data Avaliação da diversidade genética em uma amostra de acessos de cacau resistentes à doença vassoura-de-bruxa, com base em dados de RAPD e pedigree
Ronaldo Carvalho dos Santos; José Luís Pires; Uilson Vanderley Lopes; Karina Peres G. Gramacho; Acassi Batista Flores; Rita de Cássia S. Bahia; Helaine C. Cristine Ramos; Ronan Xavier Corrêa; Dario Ahnert
Genetic diversity in cocoa (Theobroma cacao L.) has been assessed based on morphological and molecular markers for germplasm management and breeding purposes. Pedigree data is available in cocoa but it has not been used for assessing genetic relatedness. The geneitic diversity of 30 clonal cocoa accessions resistant to witche´ broom disease, from the CEPEC series, were studied on the basis of RAPD data and pedigree information. Twenty of these accessions descend from the TSA-644 clone, ...
Total protein electrophoresis and RAPD fingerprinting analysis for the identification of Aeromonas at the species level Análise eletroforética de proteínas totais e marcadores de RAPD na identificação de Aeromonas ao nível de espécie
Ana Paula Longaray Delamare
Full Text Available Fifteen well-defined strains of Aeromonas of thirteen species were analyzed by SDS protein electrophoretic analysis (SDS-PAGE and random amplified polymorphic DNA analysis (RAPD. The comparison between the patterns obtained by both methods allowed differentiating all the strains. Clusters formed by the unweighted pair group method with arithmetic averages applied to protein data correlates with the genetic and biochemical information about the species. The results show that protein fingerprinting has the potential to differentiate Aeromonas species, but the low qualitative variation indicates that this technique is not efficient for the characterization of strains within a species. Conversely, RAPD fingerprinting allows the identification of strains but the high variability limits its potential as an aiding method for species identification.Quinze linhagens de Aeromonas pertencentes a treze espécies foram avaliadas através de eletroforese de proteínas totais (SDS-PAGE e segmentos de DNA amplificados ao acaso (RAPD. A comparação entre os padrões obtidos por ambos métodos permitiu diferenciar todas as linhagens. Agrupamentos formados com base nos dados protéicos mostraram relação com informações bioquímicas e genéticas a respeito das espécies. Os resultados mostraram que análises protéicas têm potencial para diferenciar espécies de Aeromonas, mas a baixa variação qualitativa indica que esta técnica não é eficiente para a caracterização entre linhagens dentro de espécies. Ao contrário, marcadores de RAPD permitem identificar linhagens, mas a alta variabilidade limita seu potencial como método auxiliar na identificação de espécies.
Faleiro Fábio G.
Full Text Available Objetivou-se neste trabalho propor uma metodologia para a utilização de marcadores RAPD como uma ferramenta auxiliar na classificação de isolados de Phytophthora spp. causadores da podridão-parda do cacaueiro (Theobroma cacao no Brasil. Existe uma necessidade constante de monitorar populações de Phytophthora spp. nas regiões cacaueiras do Brasil e a tarefa de classificação dos isolados é difícil e demorada. Com base em estudos de diversidade genética de isolados de Phytophthora capsici, P. palmivora e P. citrophthora por meio de marcadores RAPD, foram escolhidos três isolados de cada espécie como padrões e dois "primers" decâmeros mais informativos na diferenciação das espécies (OPA 13 e OPH 18. O DNA genômico dos isolados padrões e de três isolados não classificados foi extraído e amplificado, utilizando-se os dois "primers" decâmeros mais informativos. Os padrões de marcadores RAPD obtidos permitiram uma diferenciação visual clara dos isolados de cada espécie e mostraram-se úteis na classificação de isolados de Phytophthora spp. A metodologia proposta já está sendo utilizada no Centro de Pesquisas do Cacau, solucionando eventuais dúvidas resultantes da caracterização morfológica dos isolados.
张慧; 裴志东; 倪冲; 康廷国
目的:应用RAPD技术鉴别通关藤及其混淆品,并分析同种不同产地通关藤基因同源性.方法:CTAB法提取7个不同产地的通关藤及其6种混淆品的DNA,以20条随机引物进行非特异扩增.结果:随机引物285( GGG AAC CCG T)可稳定扩增不同产地通关藤药材的DNA,随机引物E01 (CCC AAG GTC C)可有效的鉴别通关藤及其混淆品.结论:RAPD法可准确鉴别正品通关藤及其混淆品,不同产地通关藤的基因具有同源性.%Objective;To identify Marsdeniae Tenacissimae Caulis and its adulterants by RAPD (random amplified polymorphic DNA) and analyze the gene homology of Marsdeniae Tenacissimae Caulis from various habitats. Methods; General DNA was isolated from Marsdeniae tenacissimae Caulis which were from seven various habitats and its six adulterants by CTAB, the twenty RAPD was used to identify them. Results; Random primer 285 (GGG AAC CCT T) could amplify the gene of Marsdeniae tenacissimae Caulis from various habitats stablely, Marsdeniae tenacissimae Caulis and its adulterants could be identified by primer E01 (CCC AAG GTC C) effectively. Conclusion;The method of RAPD can be used to identify Marsdeniae Tenacissimae Caulis and its adulterants, the gene of Marsdeniae tenacissimae Caulis from various habitats have homology.
莫小路; 曾庆钱; 黄珊珊; 蔡岳文; 王玉生; 严振
Objective:To identify the different plant resources of Herba Rabdosiae Serrae by using Random Amplified Polymorphic DNA ( RAPD ) Analysis. Methods: The Mini Spin Columns were used to extract the genomic DNA from five different plants of Herba Rabdosiae Serrae. With the DNA extracted from these plants as template,the 85 oligo nucleic acids (10 bp)as random primers,the Polymer Chain Reaction(PCR) was done and the results were analysed by electro-pharoses. Results: 12 primers were selected with polymorphism and 7 of them showed good polymorphism in RAPD map. Conclusion; RAPD method can be used to identify the plant resources of Herba Rabdosiae Serrae.%目的:用分子标记技术对中药溪黄草的基原植物分类鉴定.方法:用离心柱法提取5种溪黄草基原植物基因组DNA,用85个10 bp的寡核苷酸随机引物进行PCR(聚合酶链式反应),将扩增产物进行琼脂糖凝胶电泳,分析电泳图谱.结果:从85条随机引物中筛选出了12条多态性较好的引物,其中7个引物的扩增图谱表现较好的多态性,且重复性较好.结论:RAPD技术可以成功鉴定溪黄草药材基原.
Lu, Sheng-You; Hong, Kuo-Hsiang; Liu, Show-Ling; Cheng, Yu-Pin; Wu, Wen-Luan; Chiang, Tzen-Yuh
We used sequence variation of the atpB- rbcL intergenic spacer of cpDNA and nested clade analysis to assess the phylogeographic pattern of Michelia formosana, a species restricted to Taiwan and the Ryukyus. In total, 31 haplotypes were identified and clustered into four major chlorotypes. Genetic composition of nearly all populations was heterogeneous and paraphyletic phylogenetically. Although the apportionment of cpDNA variation hardly revealed a geographic pattern due to the coancestry of dominant sequences, some chlorotypes were restrictedly distributed. According to the patterns of clade dispersion and displacement, a reconstructed minimum spanning network revealed that historical events of past fragmentation and range expansion, associated with glaciation, may have shaped the phylogeographic patterns of M. formosana. Four possible refugia were identified: the Iriomote and Ishigaki Islands (the southern Ryukyus), Wulai (northern Taiwan), and Nanjen (southern Taiwan), on the basis of the interior positions of their haplotypes in the network and the high level of nucleotide diversity. Given insufficient time for coalescence at the cpDNA locus since the late Pleistocene recolonization, lineage sorting led to low levels of genetic differentiation among populations. In contrast, hierarchical examination of the random amplified polymorphic DNA (RAPD) data scored from six populations across three geographical regions, using an analysis of molecular variance (AMOVA), indicated high genetic differentiation both among populations (Phi(ST) = 0.471) and among regions (Phi(CT) = 0.368). An unweighted pair group method with arithmetic mean (UPGMA) tree of the RAPD fingerprints revealed that populations of two offshore islands of eastern Taiwan ( M. formosana var. kotoensis) were clustered with geographically remote populations of the Ryukyus instead of those in southern Taiwan, suggesting some historical division due to geographic barriers of the central mountain range. In
Rathore, Mangal Singh; Chikara, J; Mastan, Shaik G; Rahman, H; Anand, K G V; Shekhawat, N S
Efficient plantlet regeneration with and without intermediate callus phase was achieved for a selected genotype of Aloe vera L. which is sweet in test and used as a vegetable and source of food. Random amplified polymorphic DNA (RAPD) and inter simple sequence repeats (ISSR) marker assays were employed to evaluate genetic stability of plantlets and validate the most reliable method for true-to-type propagation of sweet aloe, among two regeneration systems developed so far. Despite phenotypic similarities in plantlets produced through both regeneration systems, the differences in genomic constituents of plantlets produced through intermediate callus phase using soft base of inflorescence have been effectively distinguished by RAPD and ISSR markers. No polymorphism was observed in regenerants produced following direct regeneration of axillary buds, whereas 80% and 73.3% of polymorphism were observed in RAPD and ISSR, respectively, in the regenerants produced indirectly from base of the inflorescence axis via an intermediate callus phase. Overall, 86.6% of variations were observed in the plantlets produced via an intermediate callus phase. The occurrence of genetic polymorphism is associated with choice of explants and method used for plantlet regeneration. This confirms that clonal propagation of sweet aloe using axillary shoot buds can be used for commercial exploitation of the selected genotype where a high degree of fidelity is an essential prerequisite. On the other hand, a high degree of variations were observed in plantlets obtained through indirect regeneration and thus cannot be used for the mass multiplication of the genotype; however, it can be used for crop improvement through induction of somaclonal variations and genetic manipulations. PMID:21887522
Full Text Available As an aim in sustainable agriculture, biological control of plant diseases has received intensive attention mainly as a response to public concern about the use of chemical fungicides in the environment. Soil Actinomycetes particularly Streptomyces spp. enhance soil fertility and have antagonistic activity against wide range of plant pathogens. To investigate for biocontrol means against the pathogen, 30 isolates of Actinomycetes have been isolated from agricultural soils of Kerman province of Iran and assayed for antagonistic activity against Botrytis allii, the agent of onion gray mold. RAPD DNA analysis has been used to determine the relatedness of active and non-active isolates based on their RAPD-PCR fingerprints. PCR amplifiable DNA samples have been isolated using the CTAB method and amplified fragments have been obtained from 5 random 10-mer primers. Different DNA fingerprinting patterns have been obtained for all of the isolates. Electrophoretic and cluster analysis of the amplification products has revealed incidence of polymorphism among the isolates. A total of 138 bands, ranging in size from 150-2800 bp, have been amplified from primers which 63.7% of the observed bands have been polymorphic. Genetic distances among different varieties have been analyzed with a UPGMA (Unweighted pair-group method, arithmetic average-derived dendrogram. Resulting dendrogram has showed from 0.65 to 0.91 similarities among varieties and divided the isolates into five major groups. Isolates which haven’t had any antagonistic activity against B. allii have been separated into a group and other isolates classified into four groups. The results indicate that RAPD is an efficient method for discriminating and studying genetic diversity of Streptomyces isolates.
Full Text Available Twenty four yardlong bean and 13 cowpea accessions were planted in the field to characterize their morphology and genetic relatedness. A randomized complete block design (RCBD with two replicationswas used. Growth habit, days to flowering, pod color, pod length, number of pods/plant, yield/plant and consumption quality were recorded. The results showed that pod length, number of pods/plant and podyield/plant among 37 accessions were highly significant differerence. Mean pod yield and pod length of 24 yardlong bean accessions were 212.1 g/plant and 48.7 cm, respectively, while mean pod yield and pod lengthof 13 cowpea accessions were 117.4 g/plant and 21.3 cm, respectively. Twenty two yardlong bean accessions exhibited indeterminate growth habit while 10 of 13 cowpea had determinate growth habit and the restsexhibited semi-determinate growth. Genetic variation and relationships among accessions were investigated based on RAPD technique. Total DNA was extracted from young leaf samples of all accessions using CTAB buffer. One hundred and twenty decamer oligonucleotide primers were screened and 5 primers (OPC-06,OPR-12, OPZ-03, OPZ-08, OPZ-13 were chosen for further evaluation. A dendrogram of genetic similarity was constructed based on 23 polymorphic bands obtained from 5 primers using UPGMA in SPSS program,which revealed separate groups between yardlong bean and cowpea. The similarity coefficient among yardlong bean and cowpea accessions ranged from 0.515 to 1.000 and 0.548 to 1.000, respectively.
Marialouis, Xavier Alexander
Introduction Global spreading of multidrug resistant strains of Escherichia coli is responsible for Urinary Tract Infection (UTI) which is a major health problem in of concern. Among the gram negative bacteria, the major contributors for UTI belongs to the family Enterobacteriaceae, which includes E. coli, Klebsiella, Citrobacter and Proteus. However, E. coli accounts for the major cause of Urinary tract infections (UTIs) and accounts for 75% to 90% of UTI isolates. Aim The main aim of this study is to analyse the phylogenetic grouping of clinical isolates of UTI E. coli. Materials and Methods In this study nearly 58 E. coli strains were isolated and confirmed through microbiological, biochemical characterization. The urine samples were collected from outpatients having symptoms of UTI, irrespective of age and sex in Tamil Nadu, India. The isolates were subjected to analyse for ESBL and AmpC β-lactamase production. To understand its genetic correlation, molecular typing was carried out using RAPD-PCR method. Results Here we noted phenotypically twenty seven isolates were positive for ESBL and seven for AmpC β-lactamase production. However, among the ESBL isolates higher sensitivity was noted for Nitrofurantoin and Cefoxitin. It is worth to note that the prevalence of UTIs was more common among female and elderly male. Phylogenetic grouping revealed the presence of 24 isolates belonged to B2 group followed by 19 isolates to group A, eight isolates to group B1 and Seven isolates to group D. Conclusion Phenotypically most of the strains were positive for ESBL and showed high sensitivity for Nitrofurantoin and cefoxitin.
The detection technology of peroxidase isoenzyme pattern and RAPD (Randomly Amplified Polymorphic DNA) analysis were used to space-flown sweet pepper variety of 87-2 carried by a recoverable satellite, and its correspondent ground control Longjiao-2. Determination of peroxidase isoenzyme revealed that the physiological activities of space-flown sweet pepper were significantly higher than those of its ground control. To detect the polymorphism of gene sequence and the difference in molecular level. 42 primers were used, among them, 38 primers amplified polymorphic products, and the amplified products from 5 primers of sweet pepper 87-2 differed from those of ground control
Manoj P. Brahmane; Krishna Mitra; Sudhanshu S. Mishra
We used random amplification of polymorphic DNA (RAPD) to generate species-specific diagnostic fragment patterns for the molecular identification of the ornamental aquarium fish species Badis badis and Dario dario. Seven arbitrary oligodecamer primers produced a total of 116 bands of which 98.23% were polymorphic. The size of the amplified products was in the range 340 bp to 2170 bp. Intraspecies genetic similarity was 0.879 ± 0.023 for B. badis and 0.840 ± 0.014 for D. dario while interspeci...
Skibsted, U.; Baggesen, Dorte Lau; Dessau, R.;
Isolates of Salmonella Enteritidis from 81 patients from Herlev Hospital or from Copenhagen County were analysed by random amplification of polymorphic DNA (RAPD), pulsed-field gel electrophoresis (PFGE) and phage-typing. Fourteen polymorphic markers from five decamer primers unambiguously placed...... (both PFGE type I) formed the fifth group. The sixth group of four isolates was not phage typeable and was PFGE type III. Forty outbreak-related isolates of phage-type 6 were resolved into three strains. No diversity of phage-type 6 was found among isolates unrelated to the outbreak. It is concluded...
Biernasiuk, Anna; Korona-Głowniak, Izabela; Grzegorczyk, Agnieszka; Malm, Anna
Cancer patients are predisposed to fungal infections caused by Candida albicans, especially to oral or respiratory tract candidiasis. The aim of this study was to estimate genetic diversity by RAPD-PCR (random amplified polymorphic DNA-polymerase chain reaction) of C. albicans isolated from upper respiratory tract of 100 patients with non-small cell lung cancer. Among 52 strains, 34 genotypes were defined. 10 clusters comprising 28 (53.85%) isolates with similarity coefficient ≥ 80% were formed. The remaining 24 (46.15%) isolates represented individual genotypes. The RAPD-PCR technique revealed genomic variability within C. albicans isolated from upper respiratory tract of the cancer patients. PMID:25371918
Diversidade genética entre híbridos de laranja-doce e tangor 'Murcott' avaliada por fAFLP e RAPD Genetic diversity among hybrids of sweet orange and 'Murcott' tangor evaluated by fAFLP and RAPD markers
Full Text Available O objetivo deste trabalho foi avaliar a diversidade genética em uma população de 148 híbridos de tangor 'Murcott' (Citrus reticulata Blanco x C. sinensis L. Osbeck e laranja 'Pêra' (C. sinensis L. Osbeck obtidos por polinização controlada, pelo uso de marcadores fAFLP e RAPD. Marcadores polimórficos (416 marcadores fAFLP e 33 RAPD foram utilizados para avaliar a similaridade genética entre os híbridos, calculada com o coeficiente Jaccard pelo método UPGMA. A consistência de cada agrupamento foi determinada pelo programa BOOD. Houve alta similaridade genética entre os parentais. A laranja 'Pêra' apresentou maior número (132 de loci em heterozigose em relação ao tangor 'Murcott' (105, corroborando a teoria de origem híbrida para a laranja-doce. Observaram-se dois grupos distintos de plantas, e um deles abrangeu 80% dos híbridos com maior similaridade com a laranja 'Pêra'. A análise bootstrap não revelou consistência estatística entre esses grupos. Marcadores fAFLP são mais eficientes na avaliação do polimorfismo, sendo indicados para seleção de indivíduos híbridos mais próximos a um dos parentais.The objective of this work was to evaluate the genetic diversity in a population of 148 hybrids of 'Murcott' tangor (Citrus reticulata Blanco x C. sinensis L. Osbeck and 'Pêra' sweet orange (C. sinensis L. Osbeck, obtained by controlled polination, using fAFLP and RAPD markers. Polymorphic markers (416 fAFLP and 33 RAPD markers were used to evaluate genetic similarity among the hybrids, calculated by the coefficient of Jaccard, using the UPGMA method. The consistency of each group was determined by software BOOD. There was high genetic similarity within the parents. 'Pêra' sweet orange had a higher number of loci in heterozygosis (132 compared to 'Murcott' tangor (105, supporting the theory of hybrid origin for sweet oranges. Two distinct groups of plants were observed: one group had 80% of the hybrids that displayed
Analía A. Lanteri; Confalonieri, Viviana A.; M. Amalia Scataglini
Después de diez años del primer registro del picudo del algodonero, Anthonomus grandis Boheman (Coleoptera, Curculionidae), en la Argentina, el insecto ha llegado a la zona algodonera del Chaco. Los estudios moleculares realizados sobre poblaciones de la Argentina, Brasil y Paraguay, y posibles poblaciones fuente de EE.UU y México, han aportado información relevante para el control de la plaga. Se aplicaron las técnicas de RAPD (Polimorfismos del ADN Amplificados al Azar) y de secuenciación d...
Lourdes Cerón González; Juan P. Legaria Solano; Clemente Villanueva Verduzco; Jaime Sahagún Castellanos
Se analizaron colecciones de cuatro especies de calabaza cultivada (Curcubita argyrosperma Huber, C. pepo L., C. moschata (Duchesne ex Lam.) Duchesne ex Poiret, y C. ficifolia Bouché) provenientes de la región Centro-Sur de México, para determinar la diversidad genética entre y dentro de las especies, y obtener las huellas genéticas correspondientes. Se utilizaron marcadores moleculares tipo RAPD (Polimorfismos en el ADN Amplificados al Azar). Se probaron 60 iniciadores y se estudió un total ...
Juan Porfirio Legaria Solano; María Elisa Alvarado Cano; Ricardo Gaspar Hernández
Se evaluó la variabilidad genética con marcadores RAPD (Polimorfismos en el ADN Amplificados al Azar) en 50 colectas de pitahaya (Hylocereus undatus Haworth, Britton and Rose) provenientes de nueve estados de México y una colecta de Colombia que se incluyó como testigo. Se detectó alta variabilidad genética (polimorfismo entre colectas de 92.5 %) en las poblaciones de pitahaya. Las huellas genéticas obtenidas permitieron la identificación individual de cada material genét...
S.M. Zakiur Rahman
Full Text Available Genetic variation is a key component for improving a stock through selective breeding programs. Randomly amplified polymorphic DNA (RAPD markers were used to assess genetic variation in three wild population of the catla carp (Catla catla Hamilton 1822 in the Halda, Jamuna and Padma rivers and one hatchery population in Bangladesh. Five decamer random primers were used to amplify RAPD markers from 30 fish from each population. Thirty of the 55 scorable bands were polymorphic, indicating some degree of genetic variation in all the populations. The proportion of polymorphic loci and gene diversity values reflected a relatively higher level of genetic variation in the Halda population. Sixteen of the 30 polymorphic loci showed a significant (p < 0.05, p < 0.01, p < 0.001 departure from homogeneity and the F ST values in the different populations indicated some degree of genetic differentiation in the population pairs. Estimated genetic distances between populations were directly correlated with geographical distances. The unweighted pair group method with averages (UPGMA dendrogram showed two clusters, the Halda population forming one cluster and the other populations the second cluster. Genetic variation of C. catla is a useful trait for developing a good management strategy for maintaining genetic quality of the species.
Sachse, Svea; Bresan, Stephanie; Erhard, Marcel; Edel, Birgit; Pfister, Wolfgang; Saupe, Angela; Rödel, Jürgen
Extended spectrum of β-lactam (ESBL) resistance of Klebsiella pneumoniae has become an increasing problem in hospital infections. Typing of isolates is important to establish the intrahospital surveillance of resistant clones. In this study, the discriminatory potential of randomly amplified polymorphic DNA and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) analyses were compared with multilocus sequence typing (MLST) by using 17 β-lactam-resistant K. pneumoniae isolates of different genotypes. MLST alleles were distributed in 8 sequence types (STs). Among ESBL strains of the same ST, the presence of different β-lactamase genes was common. RAPD band patterns also revealed 8 types that corresponded to MLST-defined genotypes in 15 out of 17 cases. MALDI-TOF analysis could differentiate 5 clusters of strains. The results of this work show that RAPD may be usable as a rapid screening method for the intrahospital surveillance of K. pneumoniae, allowing a discrimination of clonally related strains. MALDI-TOF-based typing was not strongly corresponding to genotyping and warrants further investigation. PMID:25266674
Franco Marília Caixeta
Full Text Available Marcadores RAPD (Random Amplified Polymorphic DNA foram usados para avaliar a diversidade genética entre 19 cultivares de feijão (Phaseolus vulgaris L.. Dos cento e oito locos de RAPD obtidos de 15 primers decâmeros, 70 foram polimórficos. Para estimar a distância genética foi usado o coeficiente de similaridade de Jaccard e as análises de agrupamento foram feitas pelos métodos UPGMA e Tocher. As análises de agrupamento confirmaram a ampla diversidade genética existente entre germoplasmas tropicais de feijão, separando as cultivares em dois grupos principais, correspondendo aos centros de domesticação Andino (genótipos de sementes médias e grandes e Mesoamericano (genótipos de sementes pequenas. No grupo Andino, a diversidade genética relativa foi maior do que no Mesoamericano.
Caracterización, por RAPD-PCR, de aislados de Pseudomonas aeruginosa obtenidos de pacientes con fibrosis quística RAPD-PCR characterization of Pseudomonas aeruginosa strains obtained from cystic fibrosis patients
Full Text Available OBJETIVO: Caracterizar a las cepas de P aeruginosa aisladas de lavados broncoalveolares de pacientes con fibrosis quística a lo largo de un periodo de tres años. MATERIAL Y MÉTODOS: Estudio prospectivo, de seguimiento de una población de pacientes con fibrosis quística. Se utilizó la técnica de la amplificación del ADN empleando PCR con bajas condiciones de especificidad (Random amplified polymorphic DNA, RAPD-PCR para la amplificación del ADN de cepas de P aeruginosa aisladas de lavados broncoalveolares de cinco pacientes con fibrosis quística, provenientes del Servicio de Neumología y Cirugía del Tórax del Instituto Nacional de Pediatría de la Ciudad de México, en el periodo de junio de 1996 a junio de 2002; se establecieron los patrones de amplificación de cada aislamiento, lo que permitió la identificación precisa de todas las cepas aisladas y el estudio de la epidemiología de P aeruginosa en los pacientes seleccionados con dicha enfermedad. RESULTADOS: Se definieron 18 patrones de amplificación del ADN que permitieron identificar a cada cepa de P aeruginosa aislada en las diferentes muestras de lavado broncoalveolar; no se encontró relación entre el fenotipo de P aeruginosa (mucoide o no mucoide y el genotipo de cada aislamiento, ya que cepas con fenotipos distintos mostraron patrones de amplificación semejantes; en nuestros pacientes se identificaron cepas con patrones de amplificación distintos a partir de una misma muestra, lo que sugiere la presencia de infecciones simultáneas por más de una cepa de P aeruginosa; se demostró que dos hermanos con la enfermedad compartían cepas con genotipos semejantes, lo que sugiere una contaminación cruzada entre ambos, y se demostró el aislamiento de cepas de P aeruginosa con genotipos semejantes a lo largo de los periodos estudiados. CONCLUSIONES: La identificación mediante la caracterización genotípica de las cepas de P aeruginosa aisladas de los pacientes con
Raquel dos Santos Carvalho
Full Text Available Nativo do Cerrado brasileiro e com alta variabilidade morfológica, o cajuzinho-do-cerrado (Anacardium humile St. Hill. apresenta frutos de grande aceitação pelas populações locais, os quais atraem por suas características peculiares, como tamanho, sabor único e potencial para uso sustentável por produtores e pela indústria. A produção de sementes limitada, acarretada pela baixa polinização e pela alta predação por animais e insetos, dificulta a propagação da espécie. O conhecimento da variabilidade genética do cajuzinho-do-cerrado é importante para maximizar o uso de seus recursos genéticos para futuros programas de melhoramento e de conservação da espécie. No presente trabalho, a variabilidade genética de 122 acessos de A. humile procedentes de 11 municípios (procedências do Cerrado de Goiás e Mato Grosso, foi estimada por meio de marcadores RAPD. As similaridades genéticas foram estimadas a partir da matriz binária, tendo sido processadas análises de agrupamento e de dispersão gráfica a partir da matriz de distâncias. Os iniciadores com maior expressão foram OPA11 e 08. Os dez iniciadores utilizados geraram 157 bandas, sendo 156 polimórficas (99 %, com média de 15,6 bandas/ iniciadores. Grande variabilidade dentro de municípios foi detectada, sendo o polimorfismo superior a 90 %, exceto da procedência de Jataí-GO. A distância entre acessos variou de 0,138 a 0,561, com média de 0,370, sendo os menores valores registrados entre os acessos de Mineiros-GO, e Serranópolis-GO. Os acessos de Caiapônia-GO, e Santo Antônio do Descoberto-GO, foram os mais distantes geneticamente. A dissimilaridade total entre acessos variou de 0,103 a 0,796, com médias de 0,390. Os acessos 87 e 114 de Serranópolis-GO, e Santo Antônio do Descoberto-GO, respectivamente, foram os mais distantes geneticamente, demonstrando a importância dessas procedências no enriquecendo do banco de germoplasma da espécie.
Dennis Tomalá; Johnny Chavarría; Beatriz E.A Escobar
Colossoma macropomum es una especie de la región amazónica perteneciente a la familia Characidae, la de mayor diversidad de peces de agua dulce. A nivel de laboratorio, se determinó la tasa de consumo de oxígeno (CO) en metabolismo de rutina de C. macropomum a diferentes pesos corporales: 40, 60, 90, 140 y 250 g (expresados por unidad de masa corporal) y temperatura del agua de 21, 26 y 31°C. El experimento se diseñó completamente al azar en arreglo factorial (5x3x3), utilizando un respirómet...
Santiago Rubio, Clara de
Objetivos: estudiar las alteraciones en el desarrollo dentario presentes en niños con necesidades especiales determinando cuales son las más frecuentes. Material y métodos: el estudio se realiza mediante el análisis de radiografías panorámicas de 50 pacientes elegidos al azar del Título Propio de la UCM “Especialista en atención odontológica integrada en el niño con necesidades especiales”. Resultado y conclusiones: la anomalía más frecuente es la agenesia (31%). Los pacientes con osteo...
Correlação entre heterose e divergência genética estimadas por cruzamentos dialélicos e marcadores moleculares rapd em populações de milho-pipoca Correlation between heterosis and genetic divergence estimated of diallel crosses and rapd molecular markers in populations of popcorn
Danilo Antonio Rinaldi
Full Text Available Com o objetivo de correlacionar a heterose, estimada através de cruzamentos dialélicos, com a divergência genética obtida pelo uso de marcadores moleculares RAPD, oito populações de milho-pipoca (1-PASHA, 2-PAPA, 3-PAAPC, 4-PO, 5-ZL, 6-CMS 042, 7-RS 20 e 8-CMS 43 foram intercruzadas em esquema dialélico completo, sem recíprocos, no ano agrícola de 2002/2003, gerando 28 híbridos. A avaliação dos híbridos foi realizada no ano agrícola de 2003/2004, em Londrina e Ponta Grossa, PR, em um ensaio com trinta e oito tratamentos, constituídos de vinte e oito combinações híbridas, oito parentais e duas testemunhas (IAC 112 e IAC TC01. O delineamento experimental utilizado foi o de blocos casualizados com três repetições. Foram avaliados seis caracteres: massa de grãos, capacidade de expansão, altura de planta, altura de espiga, prolificidade e florescimento feminino. Foi utilizada a técnica de RAPD para a obtenção das estimativas de distâncias genéticas entre as populações. Os resultados inferem em correlações positivas e significativas entre a divergência genética detectada pelos marcadores RAPD e massa de grãos, altura de plantas, altura de espiga e prolificidade, dos vinte e oito híbridos avaliados no dialelo em estudo. Para capacidade de expansão, florescimento e heterose percentual não foi detectada correlação significativa com a divergência genética.The objective of this study was to correlate the heterosis evaluated by diallel complete design with the genetic divergence estimated through the use of RAPD markers. Eight popcorn populations (1-PASHA, 2-PAPA, 3-PAAPC, 4-PO, 5-ZL, 6-CMS 42, 7-RS 20 and 8-CMS 43 were intercrossed on a complete diallel scheme, without reciprocal crosses, during 2002/2003 summer season, resulting in 28 hybrids. Hybrid evaluations were accomplished in the 2003/2004 summer season, at Londrina and Ponta Grossa, PR, in a trial with thirty-eight treatments, including all hybrid
Dhakshanamoorthy, Dharman; Selvaraj, Radhakrishnan; Chidambaram, Alagappan
The presence of important chemical and physical properties in Jatropha curcas makes it a valuable raw material for numerous industrial applications, including the production of biofuel. Hence, the researcher's interest is diversified to develop more and better varieties with outstanding agronomic characteristics using conventional breeding. Among these, mutation breeding is one of the best approaches to bring genetic changes in plant species. The aim of this study is to evaluate the diversity and genetic relationship among J. curcas mutants, which were obtained from different doses of gamma rays (control, 5 Kr, 10 Kr, 15 Kr, 20 Kr and 25 Kr) and EMS (1%, 2%, 3% and 4%), using RAPD marker. Among the 21 random primers, 20 produced polymorphic bands. The primers, OPM-14 and OPAW-13, produced a minimum number of bands (3) each across the ten mutants, while the primer OPF-13 produced the maximum number of bands (10), followed by the primers OPU-13, OPAM-06, OPAW-09 and OPD-05, which produced 9 bands each. The number of amplicons varied from 3 to 10, with an average of 7 bands, out of which 4.57 were polymorphic. The percentage of polymorphism ranged from 0.00 to 100 with an average of 57%. In the present study, RAPD markers were found most polymorphic, with an average polymorphism information content (PIC) value of 0.347, effective multiplex ratio (EMR) of 35.14, marker index (MI) of 14.19, resolution power (Rp) of 11.19, effective marker index (EMI) of 8.21 and genotype index (GI) of 0.36, indicating that random primers are useful in studies of genetic characterization in J. curcas mutant plants. In a dendrogram constructed based on Jaccard's similarity coefficients, the mutants were grouped into three main clusters viz., (a) control, 10 Kr, 15 Kr, 20 Kr, 2% EMS, and 3% EMS, (b) 5 Kr and 1% EMS, and (c) 25 Kr and 4% EMS mutants. Based on the attributes of the random primers and polymorphism studied, it is concluded that RAPD analysis offers a useful molecular marker
Genotype characterization of Haematobia irritans from different Brazilian geographic regions based on randomly amplified polymorphic DNA (RAPD analysis Caracterização genotípica de Haematobia irritans procedentes de diferentes regiões geográficas brasileiras baseada na análise do DNA polimórfico amplificado ao acaso (RAPD-PCR
Luciana G. Brito
Full Text Available Blood-sucking diptera are important parasites in bovine production systems, especially regarding confinement conditions. Haematobia irritans, the horn fly, is one of the most troublesome species within bovine production systems, due to the intense stress imposed to the animals. An important aspect while studying the variability within a species is the study of the geographic structure of its populations and, attempting to find out the genetic flow of Brazilian populations of horn fly, the RAPD technique, which is suited for this purpose, has been used. The use of molecular markers generated from RAPD made it possible to identify the geographic origin of samples from different Brazilian geographic regions, as well as to estimate the genotypic flow among the different Brazilian populations of the horn fly.Dípteras hematófagos são importantes parasitas dentro de sistemas de produção de bovinos, especialmente em confinamento. Haematobia irritans, a mosca-dos-chifres, é uma das espécies que maiores problemas causa em sistemas de produção de bovinos, dado ao intenso estresse que impõe aos animais. Um importante aspecto quando se estuda a variabilidade genética dentro das espécies é o estudo da estrutura geográfica destas populações. Buscando-se estimar a similaridade genotípica das diferentes populações brasileiras da mosca do chifre utilizou-se a técnica do DNA polimórfico amplificado ao acaso (RAPD-PCR, que mostrou-se eficiente para tal propósito. A utilização dos marcadores moleculares gerados através da técnica de RAPD-PCR tornou possível a identificação da origem geográfica das amostras das diferentes regiões geográficas brasileiras, assim como, estimar o fluxo genotípico entre as diferentes populações brasileiras da mosca-dos-chifres.
Marcadores moleculares RAPD e descritores morfológicos na avaliação da diversidade genética de goiabeiras (Psidium guajava L. = RAPD molecular markers and morphological descriptors in the evaluation of genetic diversity of guava (Psidium guajava L.
Aroldo Gomes Filho
Full Text Available O conhecimento da variabilidade genética e fenotípica entre diferentes acessos de goiabeiras é importante para se apoiar programas de melhoramento dessa espécie na região Norte Fluminense que carece de novas culturas capazes de gerar renda aos produtores locais. O objetivo deste trabalho foi avaliar a divergência genética entre seis cultivares e 19 acessos de goiabeiras, por meio de marcadores moleculares RAPD e características morfoagronômicas. Foram obtidas 117 marcas polimórficas, utilizando-se 28 iniciadores. Os resultados mostraram uma concordância parcial entre os métodos de agrupamentos estudados, com a formação de 12 grupos. O acesso Vita 3 e o acesso 6 foram os mais divergentes, apresentando distância genética de 0,663. A análise comparativa dos agrupamentos revelou que os marcadores RAPD e os descritores morfológicos foram eficientes para discriminação dos acessos e que houve variabilidade genética potencial para uso em Programa de Melhoramento Genético.The knowledge of the genetic and phenotypic variability among different accessions of guava is important for supporting improvement programs of this specie in northern Rio de Janeiro state, which needs new cultivars able to generate income for local farmers. This work aimed to evaluate the genetic divergence among six cultivars and 19 accessions of guava via RAPD molecular markers and morphologicalcharacteristics. One hundred and seventeen polymorphic markers were obtained from 28 primers. The results showed a partial agreement between the methods of studied groupings, with the formation of 12 groups. The accessions ‘Vita 3’and ‘6’ were the most divergent, showing genetic distance of 0.663. The comparative analysis of groupings showed that RAPD markers and morphological descriptors were effective in discriminating the accessions and to show potentialgenetic variability useful in genetic improvement programs.
Marcadores RAPD e caracteres morfoagronômicos na determinação da diversidade genética entre acessos de pimentas e pimentões RAPD markers and morphoagronomic traits in determining genetic diversity among chili peppers and sweet peppers
Fabiane Rabelo da Costa; Telma Nair Santana Pereira; Cláudia Pombo Sudré; Rosana Rodrigues
A diversidade genética existente em coleções e bancos de germoplasma pode ser estimada por meio de diversos métodos, sendo que a escolha destes depende da disponibilidade dos recursos e da precisão desejada pelo pesquisador. Neste trabalho, marcadores RAPD e caracteres morfoagronômicos foram usados para estimar a divergência genética entre 52 acessos de Capsicum spp. Um total de 57 variáveis binárias geradas pela caracterização morfoagronômica e 84 bandas polimórficas obtidas a partir da anál...
Full Text Available Abstract The aim of the present paper is to present a preliminary genetic survey of the fallow deer (Dama dama L., 1758 population of the island of Rhodes (Greece in order to verify its genetic variability. Italian population specimens were chosen as a control group because, as can be ascertained from literature, they have a very low level of variability. The analysis was carried out on hair samples obtained from each population. It was performed on a PCR modified method called RAPD which is based on the amplification of genomic DNA by using a single oligonucleotide of random sequence as a primer. The results of the analysis of the Rhodian specimens show clearly the presence of polymorphic individuals, absent in the Italian deer sampled.
Zimback Léo; Barbosa Wilson; Mori Edson Seizo; Veiga Renato Ferraz de Arruda
Foi realizada a caracterização genética das cultivares de pêssego Tropical e Douradão pelo método RAPD, em comparação com 'Dourado-1', 'Tutu', 'Maravilha', 'Rubro-sol', 'Flordaprince', 'Aurora-1' e 'Talismã' do Banco Ativo de Germoplasma de Frutas de Caroço do IAC, mantido no Núcleo de Agronomia de Capão Bonito, utilizando DNA extraído de folhas jovens. Foram obtidos 31 marcadores genéticos a partir dos primers altamente polimórficos OPAD10, OPAE04, OPAE07, OPAE09, OPAE11, OPAJ04 e OPAJ06, se...
LONG Wen-hong; XU Ming-hui; ZHANG Shu-hua
The relationship between the indica-japonica differentiation of parents and heterosis of hybridin rice was studied with 10 sterile lines, 10 maintainer lines of the sterile lines, 38 restorer lines of Dian typehybrid rice and 40 hybrids derived from the sterile lines and restorer lines. The results indicated that there wasa parabolic correlation between the indica-japonica differentiation of parents detected by RAPD markers andheterosis of hybrid rice. When certain indica-japonica differentiation of parents exists, the percentage of com-binations with positve heterosis and the mean heterosis of hybrids were high. But that didn't mean too muchdifference of parents could lead to stronger hetesosis, that meant overdiffernt parents seldom bring positve het-erotic hybrids. Suitable difference of percent of indica alleles of parents of stronger heterotic hybrid was 12 -16%.
Full Text Available The aim of the study was to evaluate the plants obtained as a result of cultivated tomato crosses with wild species meant to transfer resistance to TSWV. Six viable plants were obtained from L. esculentum x L. chilense and L. esculentum x L. peruvianum crosses after the application of in vitro embryo culture. In terms of such morphological traits as growth habit of plants, size and shape of leaves, the length and colour of internodes in branching stems, the plants displayed intermediate traits, resembling, nonetheless, the wild form. RAPD analysis with 8 primers revealed that all the hybrids had bands typical of the paternal forms. This confirms the paternal component in hybrid development. As far as the resistance to Polish TSWV isolates is concerned, two hybrids exhibited a high level of resistance, similar to negative control, three hybrids - enhanced resistance and one hybrid was susceptible to TSWV infection.
Thakur, Julie; Dwivedi, Mayank D.; Sourabh, Pragya; Uniyal, Prem L.; Pandey, Arun K.
Pittosporum eriocarpum Royle, a medicinally important taxon, is endemic to Uttarakhand region of Himalaya. It has become endangered due to over-collection and the loss of habitats. As raising plants through seeds in this plant is problematic, a reliable protocol for micropropagation using nodal explants has been developed. High shoot regeneration (95%) occurred in MS medium augmented with BA 0.4mg/l in combination IBA 0.6mg/l. In vitro regenerated shoots were rooted in MS medium supplemented with three auxins, of which 0.6 mg/l indole butyric acid proved to be the best for rooting (90%) with maximum number of roots per shoot. Thereafter, rooted plants were hardened and nearly 73% of rooted shoots were successfully acclimatized and established in the field. Start codon targeted (SCoT), inter simple sequence repeats (ISSR) and random amplified polymorphic DNA (RAPD) markers were used to validate the genetic homogeneity amongst nine in vitro raised plantlets with mother plant. DNA fingerprints of in vitro regenerated plantlets displayed monomorphic bands similar to mother plant, indicating homogeneity among the micropropagated plants with donor mother plant. The similarity values were calculated based on SCoT, ISSR and RAPD profiles which ranged from 0.89 to 1.00, 0.91 to 1.00 and 0.95 to 1.00 respectively. The dendrograms generated through Unweighted Pair Group Method with arithmetic mean (UPGMA) analysis revealed 97% similarity amongst micropropagated plants with donor mother plant, thus confirming genetic homogeneity of micropropagated clones. This is the first report on micropropagation and genetic homogeneity assessment of P. eriocarpum. The protocol would be useful for the conservation and large scale production of P. eriocarpum to meet the demand for medicinal formulations and also for the re-introduction of in vitro grown plants in the suitable natural habitats to restore the populations. PMID:27434060
宋书娟; Robert. C.Clarke; 邵宏
中国有丰富的大麻资源.目前,在我国对于大麻的研究多集中在形态和化学成分等方面,而分子遗传水平的研究至今尚未见报道.RAPD分子标记技术是适合于大麻分子水平研究的首选方法.本文以采集多年的大麻植物干标本的叶为研究材料,分别对Mg2+,dNTPs,Taq DNA酶的浓度,和退火温度和退火时间以及反应循环数等条件进行了检测,得到了适合大麻RAPD研究的理想条件.实验结果表明,用干标本来进行一定的分子遗传研究是可行的,且结果也相对稳定.%China has a great resource of Cannabis. Research on the taxonomy and morphology of Chinese Cannabis has been carried out, but so far no molecular genetic research has been published. Random amplified polymorphic DNA (RAPD) is a suitable technique for molecular genetic research on Cannabis. In this experiment,using Cannabis herbarium specimens as a source of genetic materials, the correlative conditions of the Polymerase chain reaction (PCR), (i.e., gradient density of Mg2+, dNTPs, Taq DNA polymerase, anneal temperature, anneal time and reaction cycles) were examined separately. An effective procedure for the RAPD analysis of Cannabis was obtained.
Nadezhda L Bolsheva
Full Text Available The wide variation in chromosome number found in species of the genus Linum (2n = 16, 18, 20, 26, 28, 30, 32, 36, 42, 72, 84 indicates that chromosomal mutations have played an important role in the speciation of this taxon. To contribute to a better understanding of the genetic diversity and species relationships in this genus, comparative studies of karyotypes and genomes of species within section Syllinum Griseb. (2n = 26, 28 were carried out. Elongated with 9-aminoacridine chromosomes of 10 species of section Syllinum were investigated by C- and DAPI/С-banding, CMA and Ag-NOR-staining, FISH with probes of rDNA and of telomere repeats. RAPD analysis was also performed. All the chromosome pairs in karyotypes of the studied species were identified. Chromosome DAPI/C-banding patterns of 28-chromosomal species were highly similar. Two of the species differed from the others in chromosomal location of rDNA sites. B chromosomes were revealed in all the 28-chromosomal species. Chromosomes of Linum nodiflorum L. (2n = 26 and the 28-chromosomal species were similar in DAPI/C-banding pattern and localization of several rDNA sites, but they differed in chromosomal size and number. The karyotype of L. nodiflorum was characterized by an intercalary site of telomere repeat, one additional 26S rDNA site and also by the absence of B chromosomes. Structural similarities between different chromosome pairs in karyotypes of the studied species were found indicating their tetraploid origin. RAPD analysis did not distinguish the species except L. nodiflorum. The species of section Syllinum probably originated from a common tetraploid ancestor. The 28-chromosomal species were closely related, but L. nodiflorum diverged significantly from the rest of the species probably due to chromosomal rearrangements occurring during evolution.
Muravenko, Olga V; Yurkevich, Olga Yu; Bolsheva, Nadezhda L; Samatadze, Tatiana E; Nosova, Inna V; Zelenina, Daria A; Volkov, Alexander A; Popov, Konstantin V; Zelenin, Alexander V
Karyotypes of species sects. Linum and Adenolinum have been studied using C/DAPI-banding, Ag-NOR staining, FISH with 5S and 26S rDNA and RAPD analysis. C/DAPI-banding patterns enabled identification of all homologous chromosome pairs in the studied karyotypes. The revealed high similarity between species L. grandiflorum (2n = 16) and L. decumbens by chromosome and molecular markers proved their close genome relationship and identified the chromosome number in L. decumbens as 2n = 16. The similarity found for C/DAPI-banding patterns between species with the same chromosome numbers corresponds with the results obtained by RAPD-analysis, showing clusterization of 16-, 18- and 30-chromosome species into three separate groups. 5S rDNA and 26S rDNA were co-localized in NOR-chromosome 1 in the genomes of all species investigated. In 30-chromosome species, there were three separate 5S rDNA sites in chromosomes 3, 8 and 13. In 16-chromosome species, a separate 5S rDNA site was also located in chromosome 3, whereas in 18-chromosome species it was found in the long arm of NOR-chromosome 1. Thus, the difference in localization of rDNA sites in species with 2n = 16, 2n = 30 and 2n = 18 confirms taxonomists opinion, who attributed these species to different sects. Linum and Adenolinum, respectively. The obtained results suggest that species with 2n = 16, 2n = 18 and 2n = 30 originated from a 16-chromosome ancestor. PMID:18500654
Bolsheva, Nadezhda L; Zelenin, Alexander V; Nosova, Inna V; Amosova, Alexandra V; Samatadze, Tatiana E; Yurkevich, Olga Yu; Melnikova, Nataliya V; Zelenina, Daria A; Volkov, Alexander A; Muravenko, Olga V
The wide variation in chromosome number found in species of the genus Linum (2n = 16, 18, 20, 26, 28, 30, 32, 36, 42, 72, 84) indicates that chromosomal mutations have played an important role in the speciation of this taxon. To contribute to a better understanding of the genetic diversity and species relationships in this genus, comparative studies of karyotypes and genomes of species within section Syllinum Griseb. (2n = 26, 28) were carried out. Elongated with 9-aminoacridine chromosomes of 10 species of section Syllinum were investigated by C- and DAPI/С-banding, CMA and Ag-NOR-staining, FISH with probes of rDNA and of telomere repeats. RAPD analysis was also performed. All the chromosome pairs in karyotypes of the studied species were identified. Chromosome DAPI/C-banding patterns of 28-chromosomal species were highly similar. Two of the species differed from the others in chromosomal location of rDNA sites. B chromosomes were revealed in all the 28-chromosomal species. Chromosomes of Linum nodiflorum L. (2n = 26) and the 28-chromosomal species were similar in DAPI/C-banding pattern and localization of several rDNA sites, but they differed in chromosomal size and number. The karyotype of L. nodiflorum was characterized by an intercalary site of telomere repeat, one additional 26S rDNA site and also by the absence of B chromosomes. Structural similarities between different chromosome pairs in karyotypes of the studied species were found indicating their tetraploid origin. RAPD analysis did not distinguish the species except L. nodiflorum. The species of section Syllinum probably originated from a common tetraploid ancestor. The 28-chromosomal species were closely related, but L. nodiflorum diverged significantly from the rest of the species probably due to chromosomal rearrangements occurring during evolution. PMID:25835524
Full Text Available Abstract Definition of the disease AL amyloidosis results from extra-cellular deposition of fibril-forming monoclonal immunoglobulin (Ig light chains (LC (most commonly of lambda isotype usually secreted by a small plasma cell clone. Most patients have evidence of isolated monoclonal gammopathy or smoldering myeloma, and the occurrence of AL amyloidosis in patients with symptomatic multiple myeloma or other B-cell lymphoproliferative disorders is unusual. The key event in the development of AL amyloidosis is the change in the secondary or tertiary structure of an abnormal monoclonal LC, which results in instable conformation. This conformational change is responsible for abnormal folding of the LC, rich in β leaves, which assemble into monomers that stack together to form amyloid fibrils. Epidemiology AL amyloidosis is the most common type of systemic amyloidois in developed countries with an estimated incidence of 9 cases/million inhabitant/year. The average age of diagnosed patients is 65 years and less than 10% of patients are under 50. Clinical description The clinical presentation is protean, because of the wide number of tissues or organs that may be affected. The most common presenting symptoms are asthenia and dyspnoea, which are poorly specific and may account for delayed diagnosis. Renal manifestations are the most frequent, affecting two thirds of patients at presentation. They are characterized by heavy proteinuria, with nephrotic syndrome and impaired renal function in half of the patients. Heart involvement, which is present at diagnosis in more than 50% of patients, leading to restrictive cardiopathy, is the most serious complication and engages prognosis. Diagnostic methods The diagnosis relies on pathological examination of an involved site showing Congo red-positive amyloid deposits, with typical apple-green birefringence under polarized light, that stain positive with an anti-LC antibody by immunohistochemistry and
Full Text Available O gênero Diadegma compreende espécies que são parasitóides de larvas da traça-das-crucíferas Plutella xylostella, uma das mais importantes pragas das plantas da família Brassicacea. Este gênero possui distribuição mundial. Neste trabalho, três populações de Diadegma spp. oriundas de diferentes países (Brasil, Ilha da Reunião e Malásia foram caracterizadas geneticamente por meio da técnica de RAPD-PCR. Não foi constatada variabilidade intra-populacional, no entanto o alto coeficiente de similaridade entre populações sugeriram que esses insetos poderiam pertencer a espécies diferentes.The genus Diadegma has species that are parasitoids of larvae of diamondback moth, Plutella xylostella, one of the most important pests of Brassicacea. This genus has a worldwide distribution. Representative samples of three Diadegma populations from Brazil, Reunion Island and Malaysia were characterized by RAPD-PCR. No intra-population variability was found. However, the high coefficient of genetic similarity between the populations suggests that they could belong to different species.
Assessment of genetic diversity on a sample of cocoa accessions resistant to witches' broom disease based on RAPD and pedigree data Avaliação da diversidade genética em uma amostra de acessos de cacau resistentes à doença vassoura-de-bruxa, com base em dados de RAPD e pedigree
Ronaldo Carvalho dos Santos
Full Text Available Genetic diversity in cocoa (Theobroma cacao L. has been assessed based on morphological and molecular markers for germplasm management and breeding purposes. Pedigree data is available in cocoa but it has not been used for assessing genetic relatedness. The geneitic diversity of 30 clonal cocoa accessions resistant to witche´ broom disease, from the CEPEC series, were studied on the basis of RAPD data and pedigree information. Twenty of these accessions descend from the TSA-644 clone, originated from a cross between the Upper Amazon germplasm called Scavina-6, the main source of resistance to witches' broom disease, and IMC-67. The ten remaining clones come from different sources including Amazon and Trinitario germplasm. RAPD data was collected using 16 primers and pedigree information was obtained from the International Cocoa Germplasm Database. Genetic similarities, genetic distances and coefficient of parentage were calculated using available software. Relatively low genetic diversity was observed in this germplasm set, probably because of great genetic relatedness amongst accessions studied and the poor representation of the germplasm. The TSA-644 descendants were more diverse than the other accessions used in the study. This might be due to the origin of the TSA clone, which was derived from highly divergent genotypes. Association between genetic similarities based on RAPD data and coefficient of parentage, based on pedigree data, was very low, probably due to the homogeneity of the breeding stocks and poor pedigree information. These findings are useful to cocoa breeders in planning crosses for the development of hybrid and clonal cultivars.A diversidade genética em cacau (Theobroma cacao L., embasada em dados morfológicos e em marcadores moleculares, tem sido avaliada com fins de manejo de germoplasma e uso no melhoramento genético. Dados de genealogia de cacau, embora disponíveis, não têm sido utilizados. Foi analisada a
刘平; 彭建营; 彭士琪; 周俊义; 代丽
Genetic relationships between Ziziphus jujuba and Ziziphus spinosa populations were studied using RAPD data in population genetics. 275 loci were gained among which 249 loci were polymorphic by 22 primers. The polymorphic loci percentage was 89% among Z. spinosa population while that of Z .jujuba population was 56%. 31 specific RAPD markers were detected on 3 Z.jujuba varieties and 15 Z. spinosa forms. Cluster analysis showed: when λ = 12, the forms tested were divided into 2 groups. One included Z. jujuba cv. Zanhuangdazao, Yudichuangan, Yuanshichuangan, Xiaochuangan, Qiantai No. 1, Luzhuangchuangan; the other included Z. jujuba cv. Zilingdan and 26 Z. spinosa forms. All above indicated that Z. jujuba and Z. spinosa should be regarded as two separate species.
Royal Society of Tropical Medicine and Hygiene joint meeting with Médecins Sans Frontières at Manson House, London, 20 March 2003: field research in humanitarian medical programmes. Médecins Sans Frontières interventions against kala-azar in the Sudan, 1989-2003.
Ritmeijer, Koert; Davidson, R. N.
Since 1989, Médecins Sans Frontières (MSF) has provided medical humanitarian assistance during outbreaks of visceral leishmaniasis (VL; kala-azar) in Sudan. First, in western Upper Nile in southern Sudan, where a VL epidemic occurred after the resumption of the civil war in Sudan in 1983, with an estimated 100,000 deaths. Later, MSF started interventions in eastern Upper Nile and Gedaref State. In these two endemic regions VL incidence has risen markedly since 2001, which could be the start o...
Incidencia de Inóculo y Variables Climáticas como Herramientas en la Toma de Decisiones del Manejo del Tizón de Fuego [Erwinia amylovora (Burill) Winslow et al.] Durante la Floración del Manzano [Malus sylvestris (L.) Mill. var. domestica (Borkh.) Mansf.
Manuel Rafael Ramírez-Legarreta; Juan Luis Jacobo-Cuéllar; Mario René Ávila-Marioni; Rafael Ángel Parra-Quezada; María Guadalupe Zacatenco-González; Alejandro Romo-Chacón
En los años 2006 y 2007 se determinó la presencia de Erwinia amylovora (Ea) en muestreos de 180 flores elegidas al azar en cuatro bloques de 60 árboles cada uno durante la floración del manzano cv. Golden Delicious, en la localidad de Cuauhtémoc, Chihuahua. Cada bloque representó diferente toma de decisiones para el uso de antibióticos: variables de clima, presencia de Ea en estigmas, variables de clima+Ea en estigmas y testigo. En 2008 se validaron los resultados en seis huertos del cv. Gold...