Analysis of genome survey sequences and SSR marker development for Siamese Mud Carp, Henicorhynchus siamensis, using 454 Pyrosequencing

Siamese mud carp (Henichorynchus siamensis) is a freshwater teleost of high economic importance in the Mekong River Basin. However, genetic data relevant for delineating wild stocks for management purposes currently are limited for this species. Here, we used 454 pyrosequencing to generate a partial genome survey sequence (GSS) dataset to develop simple sequence repeat (SSR) markers from H. siamensis genomic DNA. Data generated included a total of 65,954 sequence reads with average length of 264 nucleotides, of which 2.79% contain SSR motifs. Based on GSS-BLASTx results, 10.5% of contigs and 8.1% singletons possessed significant similarity (E value < 10–5) with the majority matching well to reported fish sequences. KEGG analysis identified several metabolic pathways that provide insights into specific potential roles and functions of sequences involved in molecular processes in H. siamensis. Top protein domains detected included reverse transcriptase and the top putative functional transcript identified was an ORF2-encoded protein. One thousand eight hundred and thirty seven sequences containing SSR motifs were identified, of which 422 qualified for primer design and eight polymorphic loci have been tested with average observed and expected heterozygosity estimated at 0.75 and 0.83, respectively. Regardless of their relative levels of polymorphism and heterozygosity, microsatellite loci developed here are suitable for further population genetic studies in H. siamensis and may also be applicable to other related taxa.

Assessment and rationalization of genetic diversity of Papua New Guinea taro (Colocasia esculenta) using SSR DNA fingerprinting

Taro (Colocasia esculenta) accessions were collected from 15 provinces of Papua New Guinea (PNG). The collection, totalling 859 accessions was collated for characterization and a core collection of 81 accessions (10%) was established on the basis of characterization data generated on 30 agro-morphological descriptors, and DNA fingerprinting using seven SSR primers. The selection of accessions was based on cluster analysis of the morphological data enabling initial selection of 20% accessions. The 20% sample was then reduced and rationalized to 10% based on molecular data generated by SSR primers. This represents the first national core collection of any species established in PNG based on molecular markers. The core has been integrated with core from other Pacific Island countries, contributing to a Pacific regional core collection, which is conserved in vitro in the South Pacific Regional Germplasm Centre at Fiji. The core collection is a valuable resource for food security of the South Pacific region and is currently being utilized by the breeding programmes of small Pacific Island countries to broaden the genetic base of the crop.

A high-density consensus map of barley linking DArT markers to SSR, RFLP and STS loci and agricultural traits

Background: Molecular marker technologies are undergoing a transition from largely serial assays measuring DNA fragment sizes to hybridization-based technologies with high multiplexing levels. Diversity Arrays Technology (DArT) is a hybridization-based technology that is increasingly being adopted by barley researchers. There is a need to integrate the information generated by DArT with previous data produced with gel-based marker technologies. The goal of this study was to build a high-density consensus linkage map from the combined datasets of ten populations, most of which were simultaneously typed with DArT and Simple Sequence Repeat (SSR), Restriction Enzyme Fragment Polymorphism (RFLP) and/or Sequence Tagged Site (STS) markers. Results: The consensus map, built using a combination of JoinMap 3.0 software and several purpose-built perl scripts, comprised 2,935 loci (2,085 DArT, 850 other loci) and spanned 1,161 cM. It contained a total of 1,629 'bins' (unique loci), with an average inter-bin distance of 0.7 ± 1.0 cM (median = 0.3 cM). More than 98% of the map could be covered with a single DArT assay. The arrangement of loci was very similar to, and almost as optimal as, the arrangement of loci in component maps built for individual populations. The locus order of a synthetic map derived from merging the component maps without considering the segregation data was only slightly inferior. The distribution of loci along chromosomes indicated centromeric suppression of recombination in all chromosomes except 5H. DArT markers appeared to have a moderate tendency toward hypomethylated, gene-rich regions in distal chromosome areas. On the average, 14 ± 9 DArT loci were identified within 5 cM on either side of SSR, RFLP or STS loci previously identified as linked to agricultural traits. Conclusion: Our barley consensus map provides a framework for transferring genetic information between different marker systems and for deploying DArT markers in molecular breeding schemes. The study also highlights the need for improved software for building consensus maps from high-density segregation data of multiple populations.

SSR analysis of cultivated groundnut (Arachis hypogaea L.) germplasm resistant to rust and late leaf spot diseases

Cultivated groundnut (Arachis hypogaea L.) is an agronomically and economically important oilseed crop grown extensively throughout the semi-arid tropics of Asia, Africa and Latin America. Rust (Puccinia arachidis) and late leaf spot (LLS, Phaseoisariopsis personata) are among the major diseases causing significant yield loss in groundnut. The development of varieties with high levels of resistance has been constrained by adaptation of disease isolates to resistance sources and incomplete resistance in resistant sources. Despite the wide range of morphological diversity observed in the cultivated groundnut gene pool, molecular marker analyses have thus far been unable to detect a parallel level of genetic diversity. However, the recent development of simple sequence repeat (SSR) markers presents new opportunities for molecular diversity analysis of cultivate groundnut. The current study was conducted to identify diverse disease resistant germplasm for the development of mapping populations and for their introduction into breeding programs. Twenty-three SSRs were screened across 22 groundnut genotypes with differing levels of resistance to rust and LLS. Overall, 135 alleles across 23 loci were observed in the 22 genotypes screened. Twelve of the 23 SSRs (52%) showed a high level of polymorphism, with PIC values ≥0.5. This is the first report detecting such high levels of genetic polymorphism in cultivated groundnut. Multi-dimensional scaling and cluster analyses revealed three well-separated groups of genotypes. Locus by locus AMOVA and Kruskal-Wallis one-way ANOVA identified candidate SSR loci that may be valuable for mapping rust and LLS resistance. The molecular diversity analysis presented here provides valuable information for groundnut breeders designing strategies for incorporating and pyramiding rust and late leaf spot resistances and for molecular biologists wishing to create recombinant inbred line populations to map these traits.

Caracterización molecular de 105 accesiones de cacao(Theobroma Cacao L.) utilizando 10 marcadores moleculares tipo SSR en INTA-CNIAB, Nicaragua, 2011

Se realizó la caracterización molecular de 105 accesiones de Cacao que contemplan 79 colectas nacionales, 21 híbridos de referencia y 5 materiales criollo referencia , congregados en 8 grupos de acuerdo a su ubicación geográfica al momento de la colecta; utilizando 10 marcadores moleculares tipo SSR con el fin de conocer la divers idad genética de Cacao colectado en finca de productores nicaragüenses . Se utilizó el método de extracción de ADN C TAB - Buffer, Amplificación de fragmentos de ADN mediante la técnica PCR , visualización de fragmentos de ADN en cámara electroforesis. Se identificaron 183 alelos en las 105 ac cesiones con una media de 18.3, los rangos de Heterosigocidad observada y esperada resultaron mayores para MTcCIR12 con 0.437 y 0.81 respectivamente ; el CIP y el Índice de Shannon proporcionaron los mayores valores para MTcCIR12 con 0.96 y 1.94 respectivamente . La media general del CIP e Índice de Shannon para todos los locus es de 0.87 y 1.37 respectivamente; e l análisis molecular de varianza reveló que las diferencias genéticas dentro de los grupos (85%) es mayor que las diferencias entre grupos (15%) ; se realizó el dendograma basado en las distancia de Nei’s donde revela diferencias m ínimas entre los grupos , la mayor relación genética son entre los grupos RSJ y Carazo ; en cambio para el grupo Carazo e Hibrido referencia se encontraron las mayores diferencias. E n el PCA se encontró materiales colectados que resultaron genéticamente pare cidos a los criollo s referencias, es el caso de RSJ 0211, RSJ 0 311, RSJ 0511, Bomat0510, Ji0210 ; los materiales Ji - Cuá 0104 y RSJ0411 están estrechamente relacionado . Se demostró que existe una alta diversidad genética y que existen materiales promisorios para posteriores estudios de mejora genética

Caracterizacion molecular de 52 accesiones de yuca (Manihot esculenta Crantz) usando nueve marcadores tipo SSR, CNIAB-INTA, Nicaragua

Se realizó un estudio para determinar la variabilidad genética de 35 accesiones de yuca colectadas en territorio nicaragüense, se incluyó como material de referencia 14 accesiones introducidas del Centro Internacional de Agricultura Tropical (CIAT), Colombia y 3 introducidas de Brasil, con el objetivo de identificar duplicidad de accesiones y comparar la estructura genética de las accesiones colectadas en Nicaragua. Los análisis de diversidad se obtuvieron de los datos de 9 m arcadores microsatélitestipo SSR. Se detectó un total de 47 alelos en los nueve microsatélites, el número de alelos varió de 3 a 9, con un valor medio de 5. El índice de diversidad genética fue alto de 0.61. El valor promedio del PIC fue de 0.60, demostrando que los marcadores más informativos y polimórficos fueron el SSRY 100, GA-131y GA-12, con alto poder de discriminación. El análisis molecular de varianza mostró que la mayor diferencia existe dentro del grupo, no así entre grupo. La mayor distancia genética determinada entre los grupos fue entre el grupo de Matagalpa con las Internacionales, presentando menor distancia las RAAS con las Rio San Juan, mientras que el grupo de Río San Juan y RAAS, presentaron la mayor identidad genética. El análisis de conglomerado mostró un coeficiente de correlación cofenética de 0.82 el cual agrupo ó seis grupos genéticamente idénticos. La información genética obtenida permitirá reducir las accesiones idénticas y seleccionar las de interés genético, para garantizar un manejo sostenible de los recursos que se dispone en el banco de germoplasma del Centro Nacional de Investigación Agropecuaria y Biotecnología (CNIAB) del Instituto Nicaragüense de Tecnología Agropecuaria (INTA).

Some biological features of Saprolegnia parasitica Coker - cause of fish dermatomycosis. [Translation from: Pervaya nauchnaya konferentsiya molodykh Uchenykh Biologov (Tezisy Dokladov) pp.84-86. Kiev, A.N.Ukr. SSR., 1964]

Parasitic and infectious diseases of fish, of wide distribution in fish-rearing ponds, retard to a significant extent the development of fish culture in the Ukraine. One of the diseases of fish attracting attention in connection with the general distribution of its causative agent, the fungus Saprolegnia parasitica Coker, in water-bodies of various types, appears to be dermatomycosis. The aim of this investigation is to study the conditions favouring the development of S. parasitica. Among the studied factors were water temperature and oxygen content.

The influence of constant temperature on the embryonic growth of Asellus aquaticus L. (Crustacea) [Translation from: Vestsi Akademii Navuk Belorusskoi SSR, Seriya Biyal. 1979(1) 128-130]

It is known that an adequately large amount of work has been devoted to investigations on the influence of temperature on the growth period of aquatic invertebrates. However, the action of the given factors on the basic biological characteristics of embryonic growth in crustaceans is virtually unknown. An experimental study of the effectiveness of the transformation of matter and energy during the period of embryogenesis in the isopod Asellus aquaticus L. under different constant temperatures was carried out. Specimens were collected in the quarry lakes of the Kurasovshchin zone (city-Minsk). The authors developed a quantitative analysis of the basic energetic properties of animals during one of the physiological stages at different constant temperatures, which allows one to determine the temperature range in which the expenditure of energy, at a given instance during embryonic growth, is minimised. For A. aquaticus this range is represented by the limits 10-22°C, during which the least expenditure of energy is observed between 14.5 and 18.8°C.

Respiration and the efficiency of energy during embryonic development of Asellus aquaticus L. (Crustacea) [Translation from: Vestsi Akademii Navuk Belorusskoi SSR, Seriya Biyal. 1978(5) 112-114, 1978]

Experimental studies were carried out on the transformation of matter and energy during embryonic development of the isopod Crustacea Asellus aquaticus L. under two constant temperatures. Material for the experiments was collected in the quarry lakes of the Kurasovshchin-zone (city-Minsk) at 9°C in April and at 18°C in June, 1977. The rate of consumption of oxygen per specimen by eggs and embryos of Asellus aquaticus was observed and dynamics of the basic biological characteristics during embryonic development studied.

Comparison of systems of biological indication approved during the course of the joint Anglo-Soviet investigations held under the auspices of the Institute of Hydrobiology of the Academy of Sciences of the Ukrainian SSR

In accordance with the plan for joint Anglo-Soviet scientific and technical collaboration on environmental problems, the comparative evaluation of systems of hydrobiological analysis of the surface water quality started in 1977 at the Regional Laboratory of the Severn-Trent Water Authority in Nottingham were continued in the spring of 1978. The investigations were carried out under the auspices of the Institute of Hydrobiology of the Academy of Sciences of the Ukrainian SSR. Hydrobiological and hydrochemical samples were collected by Soviet and British specialists from the Kiev reservoir and the rivers Dnieper, Sozh, Desna and Snov. The samples were processed on the expedition ships and in the Laboratory for the Hydrobiology of Small Water Bodies of the Institute of Hydrobiology of the Academy of Sciences of the Ukrainian SSR. The possible approved methods to be adopted were evaluated from the samples using the phytoperiphyton, phytoplankton, zooplankton and zoobenthos against a background of hydrochemical characteristics. The study concludes that weather conditions complicated the work on testing the systems of biological indication of water quality and made it inadvisable to use those methods of comparison which were used when similar work was carried out in Nottingham.