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  • Springer  (201)
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  • 1995-1999  (201)
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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Entomologia experimentalis et applicata 91 (1999), S. 359-368 
    ISSN: 1570-7458
    Keywords: Barbarea vulgaris ssp. arcuata ; Cruciferae ; Phyllotreta nemorum ; Chrysomelidae ; Alticinae ; flea beetle ; plant defence ; host plant range ; near-isogenic ; Y-linkage ; evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A Y-linked gene (R-gene) in the flea beetle Phyllotreta nemorum L. (Coleoptera: Chrysomelidae: Alticinae) confer the ability of larvae to survive on types of the plant Barbarea vulgaris R.Br. (Brassicaceae) which are immune to attack by susceptible conspecifics. Two near-isogenic flea beetle lines were developed. The YE-line contained the Y-linked R-gene, and male larvae from this line survived on B. vulgaris. The ST-line did not contain the gene and did not survive on the plant. The YE-line had been developed through 8–9 generations of backcrosses (YE-males with ST-females) and the two lines were considered to be isogenic except for genes located on the Y-chromosome. A single copy of the Y-linked gene is sufficient to transfer a susceptible genotype (ST) into a resistant genotype (YE) which is able to utilize a plant that is immune to attack by specimens without R-genes. The Y-linked gene had no effects on survival on other plant species tested. The gene did not have any effect on developmental times and weights of adult beetles reared on other plants than B. vulgaris. Developmental times of larvae with the Y-linked gene were longer on B. vulgaris than on normal host plants, R. sativus and S. arvensis, but the adults obtained the same size on these plant species. No trade-offs of the Y-linked gene were discovered. The results suggest that the occurrence of the Y-linked gene is a derived trait which has enabled the flea beetle to expand its host plant range. The evolution of a host shift to B. vulgaris seems not to be favoured by the presence of this single gene.
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  • 2
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    Entomologia experimentalis et applicata 91 (1999), S. 29-35 
    ISSN: 1570-7458
    Keywords: chemoreception ; deterrents ; Pieris ; Brassicaceae ; cardenolides ; host-plant selection ; evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Pieris butterflies (Lepidoptera: Pieridae) are specialist herbivores of cruciferous plants. They exploit glucosinolates, secondary plant metabolites chemotaxonomically characteristic for this plant family, as token stimuli. In addition to particular glucosinolates, some genera of the Cruciferae contain cardenolides, steroidal allelochemicals that act as potent feeding and oviposition deterrents to several Pieris species. We investigated the sensory mechanisms by which these compounds are perceived in larvae. Pieris caterpillars and many other lepidopterous species are endowed with so-called generalist deterrent receptors, that respond to a broad spectrum of secondary plant substances. In Pieris caterpillars we found a second type of deterrent chemoreceptor in maxillary styloconic taste sensilla. This neuron is very sensitive to cardenolides (threshold 0.1–0.3 μM). The generalist deterrent receptor also responds to these substances but its threshold lies at 50–100× higher concentrations. In behavioural preference experiments Pieris brassicae L. caterpillars preferred cardenolide-treated cabbage leaf discs when confronted with a choice between them and a deterrent substance that does not occur in the Brassicaceae. The cardenolides acted as potent deterrents when offered against untreated cabbage leaf discs. This demonstrates that the balance of activity elicited in the two types of deterrent chemoreceptors determines the behavioural decision.
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  • 3
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    Acta biotheoretica 47 (1999), S. 29-40 
    ISSN: 1572-8358
    Keywords: Sexual selection ; mate selection ; gamete selection ; evolution ; ploidy ; asssortative mating
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Results of an agent-based computer simulation of the evolution of diploid sexual organisms showed that several mate selection strategies confer much higher average fitness to the simulated populations, and higher evolutionary stability to the alleles coding for these strategies, than random mating. Strategies which select for 'good genes' were very successful, and so were strategies based on assortative mating. The results support the hypothesis that mating is not likely to be random in nature and that the most successful mate selection strategies are those based on assortative mating or on advantageous genes.
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  • 4
    ISSN: 1570-7458
    Keywords: Sitobion avenae ; Sitobion fragariae ; RAPD ; PCR ; microsatellites ; mtDNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A set of molecular markers to differentiate the aphid (Hemiptera: Aphidoidea) species Sitobion avenae (Fabricius) from Sitobion fragariae (Walker), is presented. These markers correspond to (1) a region of the mitochondrial DNA, (2) five species-specific RAPD banding patterns and (3) four microsatellite loci. Each of the markers was able to clearly distinguish between the species. The utility of each molecular marker is discussed. Mitochondrial DNA is best applicable to species determination and relative abundance, RAPDs to the evaluation of genetic diversity, and microsatellites to the assessment of the population genetic structure; the combined use of mtDNA with the other techniques can be of importance when the presence of hybrids is suspected, and RAPDs with microsatellites are best used together in population genetics and host preference studies.
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  • 5
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    Cellular and molecular life sciences 51 (1995), S. 454-464 
    ISSN: 1420-9071
    Keywords: Quantitative genetics ; life history ; evolution ; cladocera ; heritability ; Daphnia ; zooplankton
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Quantitative genetic techniques are powerful tools for use in understanding the microevolutionary process. Because of their size, lifespan, and ease of culture, many zooplankton species are ideal for quantitative genetic approaches. As model systems, studies of zooplankton life histories are becoming increasingly used for examination of the central paradigms of evolutionary theory. Two of the fundamental empirical questions that zooplankton quantitative genetics studies can answer are: 1) How much genetic variance exists in natural populations for life history traits? 2) What is the empirical evidence for trade-offs that permeate life history theory based on optimality approaches? A review of existing data onDaphnia indicates substantial genetic variance for body size, clutch size, and age at first reproduction. Average broad-sense heritabilities for these three characters across 19 populations of 6 species are 0.31, 0.31, and 0.34, respectively. Although there is some discrepancy between the two pertinent studies that were designed to decompose the total genetic variance into its additive and non-additive components, a crude average seems to suggest that approximately 60% of the total genetic variance has an additive basis. The existing data are somewhat inconsistent with respect to presence/absence of trade-offs (negative genetic correlations) among life history traits. A composite of the existing data seems to argue against the existence of strong trade-offs between offspring size and offspring number, between present and future reproduction, and between developmental rate and fecundity. However, there is some evidence for a shift toward more negative (less positive) covariances in more stressful environments (e.g., low food). Zooplankton will prove to be very useful in future study in several important areas of research, including the genetics and physiology of aging, the importance of genotype-environment interaction for life history traits, and the evolution of phenotypic plasticity.
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  • 6
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    Minds and machines 9 (1999), S. 309-346 
    ISSN: 1572-8641
    Keywords: language ; grammar ; syntax ; semantics ; evolution ; emergence ; brain size
    Source: Springer Online Journal Archives 1860-2000
    Topics: Computer Science , Philosophy
    Notes: Abstract It is commonly argued that the rules of language, as distinct from its semantic features, are the characteristics which most clearly distinguish language from the communication systems of other species. A number of linguists (e.g., Chomsky 1972, 1980; Pinker 1994) have suggested that the universal features of grammar (UG) are unique human adaptations showing no evolutionary continuities with any other species. However, recent summaries of the substantive features of UG are quite remarkable in the very general nature of the features proposed. While the syntax of any given language can be quite complex, the specific rules vary so much between languages that the truly universal (i.e. innate) aspects of grammar are not complex at all. In fact, these features most closely resemble a set of general descriptions of our richly complex semantic cognition, and not a list of specific rules. General principles of the evolutionary process suggest that syntax is more properly understood as an emergent characteristic of the explosion of semantic complexity that occurred during hominid evolution. It is argued that grammatical rules used in given languages are likely to be simply conventionalized, invented features of language, and not the result of an innate, grammar-specific module. The grammatical and syntactic regularities that are found across languages occur simply because all languages attempt to communicate the same sorts of semantic information.
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  • 7
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    Journal for general philosophy of science 30 (1999), S. 37-58 
    ISSN: 1572-8587
    Keywords: protein ; experimentation ; conceptual variation and selection ; evolution ; Mulder ; Liebig ; Pflüger ; Nägeli
    Source: Springer Online Journal Archives 1860-2000
    Topics: Philosophy , Nature of Science, Research, Systems of Higher Education, Museum Science
    Notes: Abstract A philosophically comprehended account is given of the genesis and evolution of the concept of protein. Characteristic of this development were not shifts in theory in response to new experimental data, but shifts in the range of questions that the available experimental resources were fit to cope with effectively. Apart from explanatory success with regard to its own range of questions, various other selecting factors acted on a conceptual variant, some stemming from a competing set of research questions, others from an altogether different field of inquiry, and still others from the external environment. These results are best explained on, hence support, an evolutionary model of the progress of experimental investigation, whose outlines are briefly discussed.
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  • 8
    ISSN: 1432-1890
    Keywords: Key words Ectomycorrhiza ; Boletus ; Amanita ; Lactarius ; Russula ; Picea abies ; RAPD ; Intra- and infraspecific variability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The application of random amplified polymorphic DNA (RAPD) analysis for the identifcation of ectomycorrhizal symbionts of spruce (Picea abies) belonging to the genera Boletus, Amanita and Lactarius at and below the species level was investigated. Using both fingerprinting [M13, (GTG)5, (GACA)4] as well as random oligonucleotide primers (V1 and V5), a high degree of variability of amplified DNA fragments (band-sharing index 65–80%) was detected between different strains of the same species, hence enabling the identification of individual strains within the same species. The band-sharing index between different species of the same genus (Boletus, Russula and Amanita) was in the range of 20–30%, and similar values were obtained when strains from different taxa were compared. Thus RAPD is too sensitive at this level of relatonship and cannot be used to align an unknown symbiont to a given taxon. We therefore conclude that RAPD is a promising tool for the identification of individual strains, and could thus be used to distinguish indigenous and introduced mycorrhizal strains from the same species in natural ecosystems.
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  • 9
    ISSN: 1432-1432
    Keywords: Escherichia coli ; RAPD ; RFLP ; Clonal theory ; Recombination
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Analysis of the Escherichia coli population by multilocus enzyme electrophoresis (MLEE) has established its clonal organization, but there is increasing evidence that horizontal DNA transfer occurs in E. coli. We have assessed the genetic structure of the species E. coli and determined the extent to which recombination can affect the clonal structure of bacteria. A panel of 72 E. coli strains from the ECOR collection was characterized by random amplified polymorphic DNA (RAPD) and restriction-fragment-length polymorphism (RFLP) of the ribosomal RNA gene (rrn) regions. These strains have been characterized by MLEE and are assumed to reflect the range of genotypic variation in the species as a whole. Statistical analysis, including factorial analysis of correspondence (FAC) and hierarchical classifications, established that the data obtained with the three genetic markers are mutually corroborative, thus providing compelling evidence that horizontal transfer does not disrupt the clonal organization of the population. However, there is a gradient of correlation between the different classifications which ranges from the highly clonal structure of 132 group strains causing extraintestinal infections in humans to the less-stringent structure of B1 group strains that came mainly from nonprimate mammals. This group (B1) appears to be the framework from which the remaining non-A group strains have emerged. These results indicate that RAPD analysis is well suited to intraspecies characterization of E. coli. Lastly, treating the RAPD data by FAC allowed description of subgroup-specific DNA fragments which can be used, in a strategy comparable to positional cloning, to isolate virulence genes.
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  • 10
    ISSN: 1437-5613
    Keywords: Key words AMOVA ; Dispersion ; Gene flow ; Genetic distance ; HOMOVA ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A survey of the genetic variability in deer mouse populations was performed using specimens collected from six different islands on a lake covering approximately 50 km2. Random amplified polymorphic DNA (RAPD) was used to measure the extent of the genetic differences in this insular system. An analysis of molecular variance (AMOVA) revealed that populations are clearly separated at this microgeographic scale (F st = 0.13863; P 〈 0.001). The homogeneity of molecular variance test (HOMOVA) indicated that within-population levels vary greatly (B p = 0.76831; P 〈 0.001). The within-population molecular variance was found to be mainly correlated with the accessibility of the islands, computed as the inverse of the geographic distance separating an island from the lakeshore (r = 0.916; P 〈 0.003).
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  • 11
    ISSN: 1572-9818
    Keywords: Date-palm ; DNA library ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A random genomic library of Tunisian date-palm varieties has been built from total cellular DNA, previously amplified according to an RAPD procedure. The resultant recombinant DNA is characterised by a size ranging from 200 to 1600 bp inserts. This DNA would constitute a large number of anonymous probes useful in Southern hybridisation experiments. It would also provide potential markers aimed at the molecular characterisation of date-palm varieties, aid the search of those associated with bayoud disease and suggest a sex determination of trees.
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  • 12
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    Plant molecular biology reporter 17 (1999), S. 171-178 
    ISSN: 1572-9818
    Keywords: Camellia sinensis ; DNA isolation ; PCR ; RAPD ; Tea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A simple procedure for DNA isolation from processed dried commercial samples of tea is described. The method involves a modified CTAB procedure employing extensive washing, use of 1% PVP to remove polyphenolics and a single phenol:chloroform extraction step. The average yield ranges from 164–494 μg/g tea sample for various market samples. The DNA obtained from 11 different brands of tea using this procedure were consistently amplifiable (using both RAPD primers as well as defined sequences as primers) and digestible with restriction endonucleases.
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  • 13
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    Journal of the history of biology 32 (1999), S. 343-383 
    ISSN: 1573-0387
    Keywords: agnosticism ; Darwinian ; evolution ; materialism ; Malthusian ; nebular hypothesis ; popularization ; professionalization ; transitional forms
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , History
    Notes: Abstract Robert Chambers and Thomas Henry Huxley helped popularize science by writing for general interest publications when science was becoming increasingly professionalized. A non-professional, Chambers used his family-owned Chambers' Edinburgh Journal to report on scientific discoveries, giving his audience access to ideas that were only available to scientists who regularly attended professional meetings or read published transactions of such forums. He had no formal training in the sciences and little interest in advancing the professional status of scientists; his course of action was determined by his disability and interest in scientific phenomena. His skillful reporting enabled readers to learn how the ideas that flowed from scientific innovation affected their lives, and his series of article in the Journal presenting his rudimentary ideas on evolution, served as a prelude to his important popular work, Vestiges of the Natural History of Creation. Huxley, an example of the new professional class of scientists, defended science and evolution from attacks by religious spokesmen and other opponents of evolution, informing the British public about science through his lectures and articles in such publications as Nineteenth Century. He understood that by popularizing scientific information, he could effectively challenge the old Tory establishment -- with its orthodox religious and political views -- and promote the ideas of the new class of professional scientists. In attempting to transform British society, he frequently came in conflict with theologians and others on issues in which science and religion seemed to contradict each other but refused to discuss matters of science with non-professionals like Chambers, whose popular writing struck a more resonant chord with working class readers.
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  • 14
    ISSN: 1420-9098
    Keywords: RAPD ; DNA polymorphism ; parental analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Previously known parent-offspring relationship for queens and her daughters of the antColobopsis nipponicus was examined using RAPD markers in order to test the reliability of this molecular technique for estimating the reproductive structure within colonies of social insects. RAPD markers from 20 oligomers successfully clustered the queen with her daughters among an artificially generated polygynous society, even when paternal information was unavailable. When information from both the queen and her sperm was included in the analysis, 20 polymorphic bands seem to be sufficient to cluster correctly the true parents to their offspring. Lack of father's information considerably decreased accuracy of the analysis. Thus, if RAPD markers are to be used to demonstrate parent-offspring relationship between individuals in the field, sperm from the queen's spermatheca should be incorporated in analysis.
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  • 15
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    Insectes sociaux 42 (1995), S. 57-69 
    ISSN: 1420-9098
    Keywords: Hindgut ; alkalinity ; evolution ; symbionts ; gut morphology
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The pH of the gut contents was measured in 52 species of higher termites (Termitidae), representing 36 genera in all four subfamilies. A statistically significant trend was shown from lower termites with low mean gut pH through to the Termitinae with higher mean gut pHs. Elevation of the pH occurred principally in the first and third proctodaeal segments, reaching values as high as 10.5 in 8 soil-feeding genera and 1 wood-feeding genus of Termitinae. Elevation of gut pH within the Termitidae appears to be independent of the general nature of the feeding substrate.
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  • 16
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    Immunogenetics 49 (1999), S. 865-871 
    ISSN: 1432-1211
    Keywords: Key words Orangutan ; MHC class I ; HLA-C ; natural killer cells ; evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  HLA-B and C are related class I genes which are believed to have arisen by duplication of a common ancestor. Previous study showed the presence of orthologues for both HLA-B and C in African apes but only for HLA-B in Asian apes. These observations suggested that the primate C locus evolved subsequent to the divergence of the Pongidae and Hominidae. From an analysis of orangutan Tengku two HLA-C-like alleles (Popy C*0101 and Popy C*0201) were defined as well as three HLA-B-like (Popy-B) alleles. By contrast, no Popy-C alleles were obtained from orangutan Hati, although three Popy-B alleles were defined. Thus an HLA-C-like locus exists in the orangutan (as well as a duplicated B locus), implying that the primate C locus evolved prior to the divergence of the Pongidae and Hominidae and is at least 12–13 million years old. Uncertain is whether all orangutan MHC haplotypes contain a C locus, as the failure to find C alleles in some individuals could be due to a mispairing of HLA-C-specific primers with certain Popy-C alleles. These results raise the possibilities that other primate species have a C locus and that the regulation of natural killer cells by C allotypes evolved earlier in primate evolution than has been thought.
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  • 17
    ISSN: 1432-203X
    Keywords: Key words Somaclonal variation ; Picea glauca ; RAPD ; Somatic embryogenesis ; Cryopreservation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Trees were regenerated from six white spruce embryogenic clones after cryopreservation for 3 and 4 years, respectively. Genetic stability was evaluated using randomly amplified polymorphic DNA (RAPD) fingerprints. Somaclonal variation was detected in some in vitro embryogenic cultures 2 and 12 months after they were re-established following cryopreservation but not in the corresponding regenerated trees. These results suggest that trees regenerated from cryopreserved cultures in subsequent years are primarily genetically stable in the genomic regions tested and that variation observed due to the in vitro culture process infrequently affects trees regenerated from normally maturing and germinating somatic embryos. However, trees regenerated from somatic embryos that matured or germinated abnormally in in vitro culture exhibited altered RAPD fragment patterns.
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  • 18
    ISSN: 1432-203X
    Keywords: Key wordsAllium sativum ; Garlic ; Genetic instability ; RAPD ; Somaclonal variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Plants were regenerated by somatic embryogenesis from long-term callus cultures derived from five garlic (Allium sativum L.) cultivars. Thirty-five of these plants were subjected to RAPD analysis. The frequency of variation was found to be cultivar dependent: approximately 1% in the two clones Solent White and California Late and around 0.35% in another three clones, Chinese, Long Keeper and Madena. Certain band changes were found in regenerants of different cultivars, suggesting the existence of a mutation-sensitive part of the garlic genome. The karyotypes of another 75 regenerants derived from the same callus cultures of three parental garlic clones were examined. Of these plants, 9.3% were found to be tetraploids, 4% aneuploid and 2.6% showed a change in the position of the secondary constriction. No association could be shown between the rate of variation for molecular and cytological characters either by comparing cultivars or examining individual regenerants.
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  • 19
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    Ecological research 10 (1995), S. 321-325 
    ISSN: 1440-1703
    Keywords: body temperature ; brood parasitism ; cuckoo ; evolution ; telemetry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Body temperatures of 11 bird species, including cuckoos, were measured in an artificial meteorological room. Ratios of change in body temperature to that in air temperature were thereby obtained for each species. Cuckoos demonstrate a remarkably high value, indicating a particularly low ability to regulate body temperature. Viewed in this light, the cuckoo's parasitic behavior is very likely an adaptation to overcome a physiological disadvantage. This in turn might be expected to reinforce delay in evolution of temperature homeostasis.
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  • 20
    ISSN: 1573-1561
    Keywords: Eriocrania cicatricella ; Eriocrania sparrmannella ; Eriocraniidae ; Lepidoptera ; sex pheromone ; EAG ; GC-EAD ; mass spectrometry ; synthesis ; evolution ; (Z)-4-hepten-2-one ; (2R)-heptan-2-ol ; (2R)-(Z)-4-hepten-2-ol
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Extracts from different body parts of adult femaleEriocrania cicatricella (Zett.) were tested for electrophysiological activity on conspecific male antennae. Extracts from the Vth abdominal segment, containing a pair of exocrine glands, elicited the largest electroantennographic response when compared to extracts of other body parts. Female extracts were analyzed by gas chromatography with simultaneous flame ionization and electroantennographic detection (EAD). The EAD active peaks were identified as (Z)-4-hepten-2-one, (2R)-heptane-2-ol, and (2R)-(Z)-4-hepten-2-ol by coinjection on a gas chromatography and by comparison of mass spectra with those of synthetic standards. In field tests, a blend of these three pheromone components was highly attractive to conspecific males, and a subtractive assay confirmed that the unsaturated alcohol is the major pheromone component, whereas no definite behavioral activity could be assigned to the ketone or the saturated alcohol. A bait containing the two alcohols withS-configuration was attractive to maleE. sparrmannella (Bosc), whereas no males ofE. cicatricella were found in these traps. The sex pheromone compounds inE. cicatricella are chemically similar to pheromones reported in Trichoptera and they are produced in homologous glands.
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  • 21
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    Journal of chemical ecology 25 (1999), S. 31-49 
    ISSN: 1573-1561
    Keywords: Chemical ecology ; evolution ; variation ; population dynamics ; community ; species interactions ; infochemical ; semiochemical ; parasitoid ; foraging behavior ; learning ; phenotypic plasticity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The marriage of chemistry with ecology has been a productive one, providing a wealth of examples of how chemicals play important roles in the loves and lives of living organisms. At first the marriage may have been a simple and monogamous one with the major scientific aim of making proximate analyses of chemically mediated, individual level interactions. But times have changed and chemical ecology is broadening, embracing different approaches and disciplines. There is, for example, increasing appreciation of variability in the systems under study and an increase in evolutionary thinking. Another promising development is greater recognition of the potential importance of chemically mediated interactions for population dynamics and for structuring communities and species coexistence. The latter is an utterly underexplored area in chemical ecology. The field of chemical ecology of insect parasitoids shows some of these promising developments. Responses of parasitoids to infochemicals are increasingly studied with an integrated approach of mechanism and function. This integration of “how” and “why” questions significantly enhances the evolutionary and ecological understanding of stimulus–response patterns. The future challenge in chemical ecology is to demonstrate how chemically mediated interactions steer ecological and evolutionary processes at all levels of ecological organization. To reach this goal there is a need for interdisciplinary collaboration among chemists and ecologists working at different levels of organization and with different approaches, with other disciplines as partners.
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  • 22
    ISSN: 1573-4943
    Keywords: Klebsiella aerogenes ; ribitol dehydrogenase ; evolution ; mutant structures
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract A mutant ribitol dehydrogenase (RDH-F) was purified from Klebsiella aerogenes strain F which evolved from the wild-type strain A under selective pressure to improve growth on xylitol, a poor substrate used as sole carbon source. The ratio of activities on xylitol (500 mM) and ribitol (50 mM) was 0.154 for RDH-F compared to 0.033 for the wild-type (RDH-A) enzyme. The complete amino acid sequence of RDH-F showed the mutations. Q60 for E60 and V215 for L215 in the single polypeptide chain of 249 amino acid residues. Structural modeling based on homologies with two other microbial dehydrogenases suggests that E60 → Q60 is a neutral mutation, since it lies in a region far from the catalytic site and should not cause structural perturbations. In contrast, L215 → V215 lies in variable region II and would shift a loop that interacts with the NADH cofactor. Another improved ribitol dehydrogenase, RDH-D, contains an A196 → P196 mutation that would disrupt a surface α-helix in region II. Hence conformational changes in this region appear to be responsible for the improved xylitol specificity.
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  • 23
    ISSN: 1573-4927
    Keywords: Akodon ; Cricetidae rodents ; genetic diversity ; biochemical polymorphism ; evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The present study involved an electrophoretic survey of 22 protein loci in 269 individuals belonging to three species of the genusAkodon, A. aff.cursor (2n=16),A. cursor (2n=14/15), andA. montensis (2n=24/25/26), collected in Eastern Brazil. The joint results of gene diversity, genetic distances, phenetic analyses, and phylogenetic trees suggested thatA. aff.cursor has recently separated fromA. cursor and that the three species have experienced a recent chromosomal divergence followed by low allozyme differentiation. These data are in agreement with their classification as sibling species.
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  • 24
    ISSN: 1573-4943
    Keywords: Ribosomal proteins ; protein sequencing ; evolution ; Haloarcula marismortui
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract The ribosomal protein HS23 from the 30S subunit of the extreme halophilicHaloarcula marismortui, belonging to the group of archaea, was isolated either by RP-HLPLC or two-dimensional polyacrylamide gel electrophoresis. The complete amino acid sequence was determined by automated N-terminal microsequencing. The protein consists of 123 residues with a corresponding molecular mass of 12,552 Da as determined by electrospray mass spectroscopy; the pI is 11.04. Homology studies reveal similarities to the eukaryotic ribosomal protein S8 fromHomo sapiens, Rattus norvegicus, Leishmania major, andSaccharomyces cerevisiae.
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  • 25
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    Plant molecular biology 29 (1995), S. 1005-1014 
    ISSN: 1573-5028
    Keywords: evolution ; genome mapping ; isozymes ; oxygen radicals ; powdery mildew
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Clones representing two distinct barley catalase genes, Cat1 and Cat2, were found in a cDNA library prepared from seedling polysomal mRNA. Both clones were sequenced, and their deduced amino acid sequences were found to have high homology with maize and rice catalase genes. Cat1 had a 91% deduced amino acid sequence identity to CAT-1 of maize and 92% to CAT B of rice. Cat2 had 72 and 79% amino acid sequence identities to maize CAT-2 and-3 and 89% to CAT A of rice. Barley, maize or rice isozymes could be divided into two distinct groups by amino acid homologies, with one group homologous to the mitochondria-associated CAT-3 of maize and the other homologous to the maize peroxisomal/glyoxysomal CAT-1. Both barley CATs contained possible peroxisomal targeting signals, but neither had favorable mitochondrial targeting sequences. Cat1 mRNA occurred in whole endosperms (aleurones plus starchy endosperm), in isolated aleurones and in developing seeds, but Cat2 mRNA was virtually absent. Both mRNAs displayed different developmental expression patterns in scutella of germinating seeds. Cat2 mRNA predominated in etiolated seedling shoots and leaf blades. Barley genomic DNA contained two genes for Cat1 and one gene for Cat2. The Cat2 gene was mapped to the long arm of chromosome 4, 2.9 cM in telomeric orientation from the mlo locus conferring resistance to the powdery mildew fungus (Erysiphe graminis f.sp. hordei).
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  • 26
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    Plant molecular biology 29 (1995), S. 1057-1070 
    ISSN: 1573-5028
    Keywords: Arabidopsis ; EF-Tu ; evolution ; gene families ; mitochondria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have characterized a second nuclear gene (tufM) in Arabidopsis thaliana that encodes a eubacterial-like protein synthesis elongation factor Tu (EF-Tu). This gene does not closely resemble the previously described Arabidopsis nuclear tufA gene, which encodes the plastid EF-Tu, and does not contain sequence elements found in all cyanobacterial and plastid tufA genes. However, the predicted amino acid sequence includes an N-terminal extension which resembles an organellar targeting sequence and shares three unique sequence elements with mitochondrial EF-Tu's, from Saccharomyces cerevisiae and Homo sapiens, suggesting that this gene encodes the Arabidopsis mitochondrial EF-Tu. Consistent with this interpretation, the gene is expressed at a higher level in flowers than in leaves. Phylogenetic analysis confirms the mitochondrial character of the sequence and indicates that the human, yeast, and Arabidopsis tufM genes have undergone considerably more sequence divergence than their cytoplasmic counterparts, perhaps reflecting a cross-compartmental acceleration of gene evolution for components of the mitochondrial translation apparatus. As previously observed for tufA, the tufM gene is present in one copy in Arabidopsis but in several copies in other species of crucifers.
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  • 27
    ISSN: 1573-5028
    Keywords: carbon fixation ; oxidative pentose phosphate pathway ; chloroplasts ; evolution ; endosymbiosis ; isoenzymes
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    Topics: Biology
    Notes: Abstract Exploiting the differential expression of genes for Calvin cycle enzymes in bundle-sheath and mesophyll cells of the C4 plant Sorghum bicolor L., we isolated via subtractive hybridization a molecular probe for the Calvin cycle enzyme d-ribulose-5-phosphate 3-epimerase (R5P3E) (EC 5.1.3.1), with the help of which several full-size cDNAs were isolated from spinach. Functional identity of the encoded mature subunit was shown by R5P3E activity found in affinity-purified glutatione S-transferase fusions expressed in Escherichia coli and by three-fold increase of R5P3E activity upon induction of E. coli overexpressing the spinach subunit under the control of the bacteriophage T7 promoter, demonstrating that we have cloned the first functional ribulose-5-phosphate 3-epimerase from any eukaryotic source. The chloroplast enzyme from spinach shares about 50% amino acid identity with its homologues from the Calvin cycle operons of the autotrophic purple bacteria Alcaligenes eutrophus and Rhodospirillum rubrum. A R5P3E-related eubacterial gene family was identified which arose through ancient duplications in prokaryotic chromosomes, three R5P3E-related genes of yet unknown function have persisted to the present within the E. coli genome. A gene phylogeny reveals that spinach R5P3E is more similar to eubacterial homologues than to the yeast sequence, suggesting a eubacterial origin for this plant nuclear gene.
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    Molecular biology reports 22 (1995), S. 139-145 
    ISSN: 1573-4978
    Keywords: chloroplast ; cyanelle ; evolution ; pre-tRNA processing ; ribozyme ; wheat germ
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract RNase P consists of both protein and RNA subunits in all organisms and organelles investigated so far, with the exception of chloroplasts and plant nuclei where no enzyme-associated RNA has been detected to date. Studies on substrate specificity revealed that cleavage by plant nuclear RNase P is critically dependent on a complete and intact structure of the substrate. No clearcut answer is yet possible regarding the order of processing events at the 5′ or 3′ end of tRNAs in the case of nuclear or chloroplast processing enzymes. RNase P from a phylogenetically ancient photosynthetic organelle will be discussed in greater detail: The enzyme from theCyanophora paradoxa cyanelle is the first RNase P from a photosynthetic organelle which has been shown to contain an essential RNA subunit. This RNA is strikingly similar to its counterpart from cyanobacteria, yet it lacks catalytic activity. Properties of the holoenzyme suggest an intermediate position in RNA enzyme evolution, with an eukaryotic-type, inactive RNA and a prokaryotic-type small protein subunit. The possible presence of an RNA component in RNase P from plant nuclei and modern chloroplasts will be discussed, including a critical evaluation of some criteria that have been frequently applied to elucidate the subunit composition of RNase P from different organisms.
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  • 29
    ISSN: 1573-5028
    Keywords: ABA-inducible genes ; coding region repeats ; embryo-specific gene family ; evolution ; Hordeum vulgare L. ; phylogenetic analysis
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    Topics: Biology
    Notes: Abstract The highly conserved Group 1 late embryogenesis abundant (Lea) genes are present in the genome of most plants as a gene family. Family members are conserved along the entire coding region, especially within the extremely hydrophilic internal 20 amino acid motif, which may be repeated. Cloning of Lea Group 1 genes from barley resulted in the characterization of four family members named B19.1, B19.1b, B19.3 and B19.4 after the presence of this motif 1, 1, 3 and 4 times in each gene, respectively. We present here the results of comparative and evolutionary analyses of the barley Group 1 Lea gene family (B19). The most important findings resulting from this work are (1) the tandem clustering of B19.3 and B19.4, (2) the spatial conservation of putative regulatory elements between the four B19 gene promoters, (3) the determination of the relative ‘age’ of the gene family members and (4) the ‘chimeric’ nature of B19.3 and B19.4, reflecting a cross-over or gene-conversion event in their common ancestor. We also show evidence for the presence of one or two additional expressed B19 genes in the barley genome. Based on our results, we present a model for the evolution of the family in barley, including the 20 amino acid motif. Comparisons of the relatedness between the barley family and all other known Group 1 Lea genes using maximum parsimony (PAUP) analysis provide evidence for the time of divergence between the barley genes containing the internal motif as a single copy and as a repeat. The PAUP analyses also provide evidence for independent duplications of Group 1 genes containing the internal motif as a repeat in both monocots and dicots.
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    Biochemical genetics 33 (1995), S. 173-181 
    ISSN: 1573-4927
    Keywords: fragile-X DNA systems ; expandable triplet repeats ; dynamic mutations ; conserved genetic domains ; evolution ; heritable disease mechanism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract A model explaining properties exhibited by fragile-X DNA systems arises from observations that time-dependent base substitutions are expressed at G-C sites but not at A–T sites (Biochem. Genet.32:383, 1994). [CGG]n sequences are classified as most sensitive to evolutionary base substitution processes involving time-dependent populating of G-C sites with enol-imine states having enhanced stability. Increased density of these states in oocyte DNA would introduce a ground-state collapse double-helix of reduced energy that would inhibit strand separation by the replicase. Evolutionarily altered G′ in CG′G triplets allows CG′G to be transcribed as CTG, an initiation codon. And this will cause reinitiation of DNA synthesis, thereby adding additional CGG units to the collapsed double helix. This situation would not occur in slower-evolving male haploid DNA that replicates frequently.
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  • 31
    ISSN: 1573-4927
    Keywords: Akodon ; Cricetidae rodents ; genetic diversity ; biochemical polymorphism ; evolution
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The present study involved an electrophoretic survey of 22 protein loci in 269 individuals belonging to three species of the genusAkodon, A. aff.cursor (2n=16),A. cursor (2n=14/15), andA. montensis (2n=24/25/26), collected in Eastern Brazil. The joint results of gene diversity, genetic distances, phenetic analyses, and phylogenetic trees suggested thatA. aff.cursor has recently separated fromA. cursor and that the three species have experienced a recent chromosomal divergence followed by low allozyme differentiation. These data are in agreement with their classification as sibling species.
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  • 32
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    Molecular biology reports 21 (1995), S. 165-167 
    ISSN: 1573-4978
    Keywords: 5S ribosomal RNA ; Harpalus rufipes ; evolution
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    Topics: Biology
    Notes: Abstract The nucleotide sequence of 5S ribosomal RNA from the beetleHarpalus rufipes was determined and compared with primary structures of other insect 5S rRNAs. Sequence differences between two beetle 5S rRNAs may represent phylogenetic markers specific for two groups of Coleoptera — Adephaga and Polyphaga. Analysis of all insect sequences using parsimony allowed us to infer a phylogenetic tree of insects, which is consistent with morphological and paleobiological data.
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  • 33
    ISSN: 1573-5028
    Keywords: Arabidopsis ; evolution ; expression ; genomic clone ; in situ hybridization ; myrosinase
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    Topics: Biology
    Notes: Abstract Myrosinase (thioglucoside glucohydrolase, EC 3.2.3.1.) is in Brassicaceae species such as Brassica napus and Sinapis alba encoded by two differentially expressed gene families, MA and MB, consisting of about 4 and 10 genes, respectively. Southern blot analysis showed that Arabidopsis thaliana contains three myrosinase genes. These genes were isolated from a genomic library and two of them, TGG1 and TGG2, were sequenced. They were found to be located in an inverted mode with their 3′ ends 4.4 kb apart. Their organization was highly conserved with 12 exons and 11 short introns. Comparison of nucleotide sequences of TGG1 and TGG2 exons revealed an overall 75% similarity. In contrast, the overall nucleotide sequence similarity in introns was only 42%. In intron 1 the unusual 5′ splice border GC was used. Phylogenetic analyses using both distance matrix and parsimony programs suggested that the Arabidopsis genes could not be grouped with either MA or MB genes. Consequently, these two gene families arose only after Arabidopsis had diverged from the other Brassicaceae species. In situ hybridization experiments showed that TGG1 and TGG2 expressing cells are present in leaf, sepal, petal, and gynoecium. In developing seeds, a few cells reacting with the TGG1 probe, but not with the TGG2 probe, were found indicating a partly different expression of these genes.
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  • 34
    ISSN: 1573-5028
    Keywords: evolution ; Chlamydomonas reinhardtii ; chloroplast ; site-specific recombination ; transcription ; transposition
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    Topics: Biology
    Notes: Abstract We have characterized two copies of a 2.4 kb DNA element that we call ‘Wendy’, in the chloroplast chromosome of Chlamydomonas reinhardtii. The two copies of Wendy reside in different single-copy regions at opposite positions in the chloroplast genome. Like many mobile DNA elements, both copies of Wendy are bordered by inverted repeats and contain several additional degenerate copies of these repeat sequences in direct or inverted orientation. In addition, four basepairs are repeated in direct orientation. Two major open reading frames (ORFs) are predicted from the DNA sequence of Wendy I. These ORFs are co-transcribed from a promoter inside the element. The deduced amino acid sequence of the larger of these ORFs shares some weak similarities with sequence motifs of transposases and integrases of other mobile elements. Wendy II appears to be altered relative to Wendy I by point mutations and small deletions and insertions which destroy the ORFs. The leader sequence of the Wendy transcript is nearly identical with the leader sequence of the rbcL transcript of C. reinhardtii, but not of C. moewusii (where the complete Wendy was also undetectable). Furthermore, both copies of Wendy are bracketed by gene clusters that are separated in C. reinhardtii but are contiguous in C. moewusii where they exist in an inverted orientation compared with C. reinhardtii. Wendy was not found in any of the completely sequenced chloroplast genomes of rice, tobacco, pine, Euglena or Marchantia, nor in any other GenBank entry. Our results suggest that Wendy has invaded C. reinhardtii after divergence from other species. Subsequent Wendy-dependent illegitimate homologous or site-specific recombination events or both may have contributed to scrambling of the C. reinhardtii chloroplast genome relative to genomes of other species.
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    Theoretical and applied genetics 90 (1995), S. 859-864 
    ISSN: 1432-2242
    Keywords: Phaseolus vulgaris ; Colletotrichum lindemuthianum ; PAL mRNA ; Cytology ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A suitable experimental model was designed with the aim of investigating the specific effect of different resistance genes in the Phaseolus vulgaris — Colletotrichum lindemuthianum interaction. The four resistance genes examined were chosen because they confer a different phenotype (resistance or susceptibility) to the lines carrying them when challenged by a range of C. lindemuthianum races. These different resistance genes were introgressed independently into the same susceptible recipient line. The isogenicity of the five near-isogenic lines (NILs) thus obtained (four resistant lines, one susceptible line = recipient line) was assessed by a RAPD analysis. The hypersensitive reaction occurred at the same time after infection, whatever the resistance gene present, when the NILs were challenged by the avirulent race 9 of the pathogen. In contrast, the pathogen development was arrested more or less rapidly in the different NILs. At the first stages of the infection process, the transcripts encoding phenylalanine ammonia-lyase were accumulated to a different extent in the different resistant NILs but always to a higher level than in the susceptible recipient line. These results suggest that the different resistance genes operate through more than one way in the production of defense factors.
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    Theoretical and applied genetics 90 (1995), S. 1049-1055 
    ISSN: 1432-2242
    Keywords: Oryza sativa ; Rice ; Genetic resources ; RAPD ; Molecular markers ; Cluster analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A set of accessions of Oryza sativa from the International Rice Research Institute (Philippines) that included known and suspected duplicates as well as closely related germplasm has been subjected to RAPD analysis. The number of primers, the number of polymorphic bands and the total number of bands were determined that will allow the accurate discrimination of these categories of accessions, including the identification of true and suspected duplicates. Two procedures have been described that could be employed on a more general basis for identifying duplicates in genetic resources collections, and further discussion on the values of such activities is presented.
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  • 37
    ISSN: 1432-2242
    Keywords: RAPD ; Sugarcane ; Embryogenic callus ; Genetic transformation ; Somaclonal variation
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    Topics: Biology
    Notes: Abstract Random amplified polymorphic DNA (RAPD) analysis using 10-mer oligonucleotide primers efficiently differentiated sugarcane cultivars and proved suitable for detecting gross genetic change such as that which can occur in sugarcane subjected to prolonged tissue culture, for example in protoplast-derived callus. However, RAPD analysis was not sufficiently sensitive to detect smaller genetic changes that occur during sugarcane genetic transformation. The length of DNA scored for polymorphism per primer averaged 13.2 kb, or 0.0001% of the typical sugarcane genome size of 1.2 × 107 kb (2C). RAPD analysis of sugarcane plants regenerated from embryogenic callus revealed very few polymorphisms, indicating that gross genetic change is infrequent during this tissue culture procedure, although epigenetic effects result in transient morphological changes in regenerated plants. More sensitive variations on the RAPD technique may increase the practicality of DNA-based screening of regenerated plant lines to reveal somaclonal variants.
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  • 38
    ISSN: 1432-2242
    Keywords: Brassica rapa ; RFLP ; RAPD ; QTL ; Palmitic acid
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    Topics: Biology
    Notes: Abstract F2 progeny (105 individuals) from the cross Jo4002 x Sv3402 were used to identify DNA markers associated with palmitic-acid content in spring turnip rape (Brassica rapa ssp. oleifera). QTL mapping and ANOVA analysis of 140 markers exposed one linkage group with a locus controlling palmitic-acid content (LOD score 27), and one RAPD (random amplified polymorphic DNA) marker, OPB-11a, closely linked (1.4 cM) to this locus. Palmitic-acid content in the 62 F2 plants with the visible allele of marker OPB-11a was 8.45 ±3.15%, while that in the 24 plants without it was 4.59 ±0.97%. As oleic-acid concentration is affected by a locus on the same linkage group as the palmitic-acid locus, this locus probably controls the chain elongation from palmitic acid to oleic acid (through stearic acid). Marker OPB-11a may be used in future breeding programs of spring turnip rape to simplify and hasten the selection for palmitic-acid content.
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    Theoretical and applied genetics 91 (1995), S. 647-654 
    ISSN: 1432-2242
    Keywords: Lens ; RAPD ; Cluster analysis ; Diversity ; Phylogeny
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract RAPD markers were used to distinguish between six different Lens taxa, representing cultivated lentil and its wild relatives. Twenty-four arbitrary sequence 10-mer primers were identified which revealed robust and easily interpretable amplification-product profiles. These generated a total of 88 polymorphic bands in 54 accessions and were used to partition variation within and among Lens taxa. The data showed that, of the taxa examined, ssp. orientalis is most similar to cultivated lentil. L. ervoides was the most divergent wild taxon followed by L. nigricans. The genetic similarity between the latter two species was of the same magnitude as between ssp. orientalis and cultivated lentil. In addition, species-diagnostic amplification products specific to L. odemensis, L. ervoides and L. nigricans were identified. These results correspond well with previous isozyme and RFLP studies. RAPDs, however, appear to provide a greater degree of resolution at a sub-species level. The level of variation detected within cultivated lentils suggests that RAPD markers may be an appropriate technology for the construction of genetic linkage maps between closely related Lens accessions.
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    Theoretical and applied genetics 90 (1995), S. 27-32 
    ISSN: 1432-2242
    Keywords: RAPD ; Genetic diversity ; Vicia faba L ; Germ plasm ; Gene pools
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Broadening of the genetic base and systematic exploitation of heterosis in faba bean requires reliable information on the genetic diversity in the germ plasm. Three groups of faba bean inbred lines were examined by means of RAPDs (random amplified polymorphic DNAs) assays: 13 European small-seeded lines, 6 European large-seeded lines, and 9 Mediterranean lines. Out of 59 primers, 35 were informative and yielded 365 bands, 289 of which were polymorphic with a mean of 8.3 bands per primer. Monomorphic bands were omitted from the analyses and genetic distances (GD) were estimated via the coefficient of Jaccard. The mean GD among the European small-seeded lines was significantly greater than those among the lines of the other two groups. Repeatability of GD estimates was high. Cluster (UPGMA) and principal coordinate analyses identified European small-seeded lines and Mediterranean lines as distinct groups with European large-seeded lines located in between. The results are in harmony with published archaeobotanical findings. We conclude that RAPDs are useful for classification of germ plasm and identification of divergent heterotic groups in faba bean.
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  • 41
    ISSN: 1432-2242
    Keywords: Leaf rust ; RFLP ; RAPD ; Wheat ; Agropyron elongatum
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    Notes: Abstract The objective of this study was to identify molecular markers linked to the wheat leaf rust resistance gene Lr24 derived from Agropyron elongatum (3DL/3Ag translocation). Two near isogenic lines (NILs), ‘Arina’ and Lr24/7 * “Arina”, were screened for polymorphism at the DNA level with 115 RFLP probes. Twenty-one of these probes map to the homoeologous group 3. In addition, 360 RAPD primers were tested on the NILs. Six RFLP probes showed polymorphism between the NILs, and 11 RAPD primers detected one additional band in the resistant NIL. The genetic linkage of the polymorphic markers with Lr24 was tested on a segregating F2 population (150 plants) derived from a cross between the leaf rust resistant Lr24/7 * “Arina” and the susceptible spelt (Triticum spelta) variety ‘Oberkulmer’. All 6 RFLP markers were completely linked to Lr24: one was inherited as a codominant marker (PSR1205), one was in coupling phase (PSR1203) and 4 were in repulsion phase (PSR388, PSR904, PSR931, PSR1067) with Lr24. The localization of these probes on chromosome 3D was confirmed by nulli-tetrasomic analysis. Distorted genotypic segregation was found for the Codominant RFLP marker PSR1205. This distortion can be explained by the occurrence of hemizygous plants. One of the 11 RAPD markers (OPJ-09) also showed complete linkage to theLr24 resistance gene. The polymorphic RAPD fragment was cloned and sequenced. Specific primers were synthesized, and they produced an amplification product only in the resistant plants. This specific marker allows a reliable and rapid screening of a large number of genotypes in practical breeding. Analysis of 6 additional lines containing Lr24 revealed that 3 lines have a smaller chromosomal segment of A. elongatum than lines derived from ‘Agent’, a commonly used gene donor for the Lr24 resistance gene.
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  • 42
    ISSN: 1432-2242
    Keywords: Key words Ascochyta lentis ; Lens culinaris ssp. culinaris ; Bulked segregant analysis ; Resistance genes ; RAPD ; QTL analysis
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Abstract  Foliar resistance to Ascochyta lentis is controlled at a single major locus by a dominant gene (AbR 1 ) in the lentil accession ILL5588 (cv ‘Northfield’). Flanking RAPD markers that are closely linked to the resistance locus in coupling phase were identified by bulked segregant analysis. Out of 261 decanucleotide primers screened 7 produced a polymorphic marker that segregated with the resistance locus, and all markers were found to exist within a single linkage group. Five of the seven RAPD markers were within 30 cM of the resistance locus. Log likelihood analysis for detecting QTL associated with the foliar resistance revealed that a single narrow peak accounted for almost 90% of the variance of resistance between the bulks. Preliminary mapping in an F3 population revealed that the closest flanking markers were approximately 6 and 14 centiMorgans (cM) away from the resistance locus. These markers should be useful for the discrimination of resistant germplasm through marker-assisted selection in future breeding programmes and represent the first essential step towards the map-based cloning of this resistance gene.
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    Theoretical and applied genetics 99 (1999), S. 58-64 
    ISSN: 1432-2242
    Keywords: Key words Genetic map ; RFLP ; AFLP ; RAPD ; SAMPL ; Daucus carota L. ssp. sativus
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Abstract  A 109-point linkage map consisting of three phenotypic loci (P 1, Y 2, and Rs), six restriction fragment length polymorphisms (RFLPs), two random amplified polymorphic DNAs (RAPDs), 96 amplified fragment length polymorphisms (AFLPs), and two selective amplification of microsatellite polymorphic loci (SAMPL) was constructed for carrot (Daucus carota L. ssp. sativus; 2n=2x=18). The incidence of polymorphism was 36% for RFLP probes, 20% for RAPD primers, and 42% for AFLP primers. The overall incidence of disturbed segregation was 18%. Linkage relationships at a LOD score of 4.0 and θ=0.25 indicated 11 linkage groups. The total map length was 534.4 cM and the map was clearly unsaturated with markers spaced at 4.9 cM. AFLP P6B15 was 1.7 cM from P 1, AFLP P1B34 was 2.2 cM from Y 2, and AFLP P3B30XA was 8.1 cM from Rs.
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    Theoretical and applied genetics 91 (1995), S. 869-875 
    ISSN: 1432-2242
    Keywords: Genetic map ; Linkage ; Eucalypts ; RFLP ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract An integrated genetic linkage map for E. nitens was constructed in an outbred three-generation pedigree. Analysis of 210 RFLP, 125 RAPD and 4 isozyme loci resulted in 330 markers linked in 12 linkage groups covering 1462 cM (n=11 in eucalypts). The 12th linkage group is comprised of only 5 markers and will probably coalesce with another linkage group when further linked loci are located. Co-dominant RFLP loci segregating in both parents were used to integrate linkages identified in the male and female parents. Differences in recombination frequencies in the two parents were observed for a number of pairs of loci, and duplication of sequences was identified both within and between linkage groups. The markers were distributed randomly across the genome except for the RFLPs in linkage group 10 and for some loci showing segregation distortion, which were clustered into three regions of the map. The use of a large number of co-dominant RFLP loci in this map enables it to be used in other pedigrees of E. nitens and forms a basis for the detection and location of QTL in E. nitens and other eucalypt species.
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    Theoretical and applied genetics 90 (1995), S. 853-858 
    ISSN: 1432-2242
    Keywords: Microseris ; RAPD ; QTL ; Trichomes Gene regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Segregation for 289 random amplified polymorphic DNA markers (RAPDs) has been determined in 106 F2 plants of an interspecific hybrid (H27) between Microseris douglasii (strain B14) and M. bigelovii (C94). Multicelluar trichomes (“type D”, specific for Microseris) occur on the leaf teeth of early vegetative rosettes of the B14 parent and on the leaf blades of later rosettes in both parents. Trichomes on the leaf blades appear earlier and eventually more densely in B14. Segregation for trichome appearance is quantitative and strongly transgressive in the F2 hybrid. Cosegregation between RAPDs and trichome phenotypes combined with linkage data have revealed a main gene (“quantitative trait locus A”, QTL-A) with a pleiotropic effect on all trichome characters and two unlinked additive modifiers (QTL-B, QTL-C). Alleles of both modifiers reduce the main gene effect in each parent. Their recombination explains the occurrence of plants with transgressive phenotypes in the hybrid offspring. Additional QTLs affecting trichomes are at and below the level of statistical significance.
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  • 46
    ISSN: 1432-2242
    Keywords: RAPD ; Pseudo-testcross ; Eucalyptus ; QTL ; Vegetative propagation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have extended the combined use of the “pseudo-testcross” mapping strategy and RAPD markers to map quantitative trait loci (QTLs) controlling traits related to vegetative propagation in Eucalyptus. QTL analyses were performed using two different interval mapping approaches, MAPMAKER-QTL (maximum likelihood) and QTL-STAT (non-linear least squares). A total of ten QTLs were detected for micropropagation response (measured as fresh weight of shoots, FWS), six for stump sprouting ability (measured as # stump sprout cuttings, #Cutt) and four for rooting ability (measured as % rooting of cuttings, %Root). With the exception of three QTLs, both interval-mapping methods yielded similar results in terms of QTL detection. Discrepancies in the most likely QTL location were observed between the two methods. In 75% of the cases the most likely position was in the same, or in an adjacent, interval. Standardized gene substitution effects for the QTLs detected were typically between 0.46 and 2.1 phenotypic standard deviations (σp), while differences between the family mean and the favorable QTL genotype were between 0.25 and 1.07 (σp). Multipoint estimates of the total genetic variation explained by the QTLs (89.0% for FWS, 67.1 % for#Cutt, 62.7% for %Root) indicate that a large proportion of the variation in these traits is controlled by a relatively small number of major-effect QTLs. In this cross, E. grandis is responsible for most of the inherited variation in the ability to form shoots, while E. urophylla contributes most of the ability in rooting. QTL mapping in the pseudo-testcross configuration relies on withinfamily linkage disequilibrium to establish marker/trait associations. With this approach QTL analysis is possible in any available full-sib family generated from undomesticated and highly heterozygous organisms such as forest trees. QTL mapping on two-generation pedigrees opens the possibility of using already existing families in retrospective QTL analyses to gather the quantitative data necessary for marker-assisted tree breeding.
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  • 47
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    Theoretical and applied genetics 98 (1999), S. 657-663 
    ISSN: 1432-2242
    Keywords: Key words Cicer ; Species relationships ; DNA fingerprinting ; RAPD ; Chickpea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Random amplified polymorphic DNA markers were used to distinguish between nine different Cicer taxa representing the cultivated chickpea and eight other related annual wild species. Of the 75 random10-mer primers tested, only 8 amplified genomic DNA across all the species. A total of 115 reproducibly scorable RAPD markers were generated, all except 1 polymorphic, and these were utilized to deduce genetic relationships among the annual Cicer species. Four distinct clusters were observed and represented C. arietinum, C. reticulatum and C. echinospermum in first cluster followed by C. chorassanicum and C. yamashitae in the second cluster, while C. pinnatifidum, C. judaicum and C. bijugum formed the third cluster. Cicer cuneatum did not cluster with any of the species and was most distantly placed from the cultivated species. Except for the placement of C. chorassanicum and C. yamashitae, deduced species’ relationships agreed with previous studies. In addition, species-diagnostic amplification products specific to all the nine species were identified. The results clearly demonstrate a methodology based on random-primed DNA amplification that can be used for studying Cicer phylogeny and chickpea improvement.
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  • 48
    ISSN: 1432-2242
    Keywords: Key words Vicia faba ; Genetic map ; Trisomics ; RAPD ; Seed-protein genes ; QTLs
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Seven F2 families of faba bean descendent from plants trisomic for chromosomes 3, 4, 5 and 6 were analyzed for isozyme markers and two of these were also studied for morphological and RAPD markers and seed-protein genes. Linkage analysis revealed 14 linkage groups, 8 of which were unambiguously assigned to specific chromosomes. Several QTLs for seed weight were identified, the most important of which, located on chromosome 6, explained approximately 30% of the total phenotypic variation. Comparison of results from Vicia faba with the maps of the related species Pisum sativum L. and Cicer arietinum L. revealed one possible new case of linkage conservation. A composite linkage analysis based on 42 markers analyzed in this and previous studies, where line Vf 6 was also used as the female parental, allowed the new assignment of previously independent linkage groups and/or markers to specific chromosomes. Thus, the number of linkage groups was reduced to 13, each comprising an increased number of markers. No contradictory results were detected, indicating the suitability of the statistical procedure and methodology used so far in the development of the map of this species.
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  • 49
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    Theoretical and applied genetics 99 (1999), S. 147-156 
    ISSN: 1432-2242
    Keywords: Key words Capsicum ; Diagnostic markers ; Genetic diversity ; Germplasm ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Germplasm characterization is an important link between the conservation and utilization of plant genetic resources. A total of 134 accessions from six Capsicumspecies maintained at the Asian Vegetable Research and Development Center were characterized using 110 randomly amplified polymorphic DNA (RAPD) markers. Ten pairs of potentially duplicated accessions were identified. Multidimensional scaling analysis of the genetic distances among accessions resulted in clustering corresponding to a previous species assignment except for six accessions. Diagnostic RAPDs were identified which discriminate among the Capsicumspecies. The diagnostic markers were employed for improved taxonomic identification of accessions since many morphological traits used in the identification of Capsicumare difficult to score. Three Capsicumaccessions, misclassified based on morphological traits, were reassigned species status based on diagnostic RAPDs. Three accessions, not previously classified, were assigned to a species based on diagnostic RAPDs. Definitive conclusions about the species assignment of three other accessions were not possible. The level of diversity between Capsicum annuumaccessions from the genebank and the breeding program were compared and no differences were observed either for RAPD variation or diversity. The utilization of genetic resources as a source of variance for useful traits in the breeding program may be the reason for the similarity of these two groups.
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  • 50
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    Theoretical and applied genetics 99 (1999), S. 1061-1067 
    ISSN: 1432-2242
    Keywords: Key words Native American maize ; RAPD ; Genetic relationships ; Reproducibility ; Geography and evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Genetic variation among 15 accessions of Native American maize from the Great Plains was investigated using random amplified polymorphic DNA (RAPD). RAPDs revealed very high levels of polymorphism among accessions. Banding patterns ranged in percentage polymorphism from 46.7% to 86.2% with an overall mean of 70.7% for the primers analyzed. The construction of genetic relationships using cluster analysis and principal coordinates analysis revealed that RAPDs are successful in confirming hypothesized relationships and in identifying misclassified specimens. Furthermore, the phenogram fails to reveal a strong correspondence between genetic relationships and the geographical position of Native Americans prior to contact. This provides support for the hypothesis that multiple introductions of maize into the Great Plains via trade may have resulted in the great morphological variation found among accessions in the region. Based on these data, it is unlikely that a separate Great Plains race of maize can be distinguished. In general, we conclude that RAPDs are potentially very useful in organizing seed collections and understanding intraspecific genetic differentiation.
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  • 51
    ISSN: 1432-2242
    Keywords: Key words Bs2 resistance gene ; Pepper ; RAPD ; AFLP ; Positional cloning
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    Topics: Biology
    Notes: Abstract  The Bs2 resistance gene of pepper confers resistance against the bacterial pathogen Xanthomonas campestris pv. vesicatoria. As a first step toward isolation of the Bs2 gene, molecular markers tightly linked to the gene were identified by randomly amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP) analysis of near-isogenic lines. Markers flanking the locus were identified and a high-resolution linkage map of the region was developed. One AFLP marker, A2, was found to cosegregate with the locus, while two others, F1 and B3, flank the locus and are within 0.6 cM. Physical mapping of the A2 and F1 markers indicates that these markers may be within 150 kb of each other. Together, these results indicate that the Bs2 region may be cloned either by chromosome walker or landing. The linked markers were also used to characterize gamma-irradiation-induced mutants at the Bs2 locus.
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  • 52
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    Theoretical and applied genetics 90 (1995), S. 166-172 
    ISSN: 1432-2242
    Keywords: Ribes nigrum ; RAPD ; Genetic variation
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    Topics: Biology
    Notes: Abstract Ribes nigrum germplasm was screened for random amplified polymorphic DNA (RAPD) markers. Fiftyfour markers were identified which generated individual fingerprints for each of 21 cultivars. Genetic variation within R. nigrum germplasm, as detected by RAPDs, demonstrated that the genetic basis for improvement of blackcurrant is narrower than would be expected by the analysis of parentage.
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  • 53
    ISSN: 1432-2242
    Keywords: Wheat ; Rye ; RAPD ; PCR ; In situ hybridization ; Dispersed repeat
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    Topics: Biology
    Notes: Abstract Bulk segregant analysis was used to obtain a random amplified polymorphic DNA (RAPD) marker specific for the rye chromosome arm of the 1BL.1RS translocation, which is common in many high-yielding bread wheat varieties. The RAPD-generated band was cloned and end-sequenced to allow the construction of a pair of oligonucleotide primers that PCR-amplify a DNA sequence only in the presence of rye chromatin. The amplified sequence shares a low level of homology to wheat and barley, as judged by the low strength of hybridization of the sequence to restriction digests of genomic DNA. Genetic analysis showed that the amplified sequence was present on every rye chromosome and not restricted to either the proximal or distal part of the 1RS arm. In situ hybridization studies using the amplified product as probe also showed that the sequence was dispersed throughout the rye genome, but that the copy number was greatly reduced, or the sequence was absent at both the centromere and the major sites of heterochromatin (telomere and nucleolar organizing region). The probe, using both Southern blot and in situ hybridization analyses, hybridized at a low level to wheat chromosomes, and no hybridizing restriction fragments could be located to individual wheat chromosomes from the restriction fragment length polymorphism (RFLP) profiles of wheat aneuploids. The disomic addition lines of rye chromosomes to wheat shared a similar RFLP profile to one another. The amplified sequence does not contain the RIS 1 sequence and therefore represents an as yet undescribed dispersed repetitive sequence. The specificity of the amplification primers is such that they will provide a useful tool for the rapid detection of rye chromatin in a wheat background. Additionally, the relatively low level of cross-hybridization to wheat chromatin should allow the sequence to be used to analyse the organization of rye euchromatin in interphase nuclei of wheat lines carrying chromosomes, chromosome segments or whole genomes derived from rye.
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  • 54
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    Theoretical and applied genetics 90 (1995), S. 767-770 
    ISSN: 1432-2242
    Keywords: Solanum melongena ; Insanum ; RAPD ; Interrelationships
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    Topics: Biology
    Notes: Abstract RAPD analysis was carried out on 52 accessions of Solanum melongena (eggplant) and related weedy forms known as “insanum”. Twenty-two primers amplified 130 fragments. Solanum melongena exhibited 117 of the fragments, all of which were also present in insanum. Insanum displayed an additional 13 fragments not found in S. melongena. Overall, the insanum accessions were more diverse than those of S. melongena. The calculated similarity between them was 0.947. The RAPD results were closely concordant with the results of an electrophoretic isozyme survey performed on the same accessions. The concordance of the results shows that even though S. melongena and insanum are highly diverse morphologically, it is no longer appropriate to distinguish them taxonomically.
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  • 55
    ISSN: 1432-2242
    Keywords: Allozymes ; Chloroplast DNA ; RAPD ; Genetic variation ; Abies
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    Topics: Biology
    Notes: Abstract Allozyme, chloroplast (cpDNA) and random amplified polymorphic DNA (RAPD) markers have been used to estimate genetic and taxonomic relationships among different populations of Abies alba and the relic population of A. nebrodensis. Twelve isozyme gene loci, as well as restriction fragment length polymorphism (RFLP) at cpDNA spacer regions between t-RNA genes were analysed. Moreover, a set of 60 random sequence 10-mer primers were tested. Over all isozyme loci, evident differences in allele frequencies among A. nebrodensis and A. alba populations were found, particularly at 2 loci, phosphoglucose isomerase (Pgi-a) and shikimate dehydrogenase (Skd-a). More than 10% of the total genetic diversity was due to differences among populations. High values of genetic distances among populations were also found. Out of the 60 primers tested, 12 resulted in a polymorphic banding pattern both within and among populations. A total of 84 RAPD fragments were produced by the 12 selected primers. A phenogram of relationships among populations was constructed based on RAPD band sharing: the differentiation of the A. nebrodensis population was evident. The analysis of molecular variance (AMOVA) was used to apportion the variation among individuals within populations and among populations. There was considerable variation within each population: even so, genetic divergence was found among populations. This pattern of genetic variation was very different from that reported for inbred species. Identical cpDNA amplification and restriction patterns were observed among all the individuals sampled from the populations. Taken together, the results of allozyme and RAPDs show a clear differentiation among A. nebrodensis and A. alba populations and provide support for their classification into two different taxonomic groups.
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  • 56
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    Theoretical and applied genetics 91 (1995), S. 142-149 
    ISSN: 1432-2242
    Keywords: DNA fingerprint ; Molecular markers ; Picea mariana ; Picea rubens ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Species-specific molecular markers were designed to assist in the identification of closely related black spruce (Picea mariana [B.S.P.] Mill.) and red spruce (P. rubens Sarg.) in northeastern North America. Trees from six provenances of black spruce and three provenances of red spruce were sampled from outside the sympatric zone. They were first classified using a composite index of five qualitative morphological traits. The species-specific genetic markers were developed using random amplified polymorphic DNAs (RAPD) and a combination of bulk sample and individual tree analyses. Each species bulk sample was constructed from DNAs obtained from 12 trees that were from outside the sympatric zone and showed a morphological composite index specific of each species. A total of 161 primers were screened with the bulk samples. From these, 52 primers showing segregating fingerprints were further screened with the individual trees. Most of the markers observed were shared by the two species, and there was less diversity in P. rubens. A small number of markers were found to be monomorphic or nearly monomorphic and specific to either P. mariana or P. rubens. These markers remained species-specific when F1 progenies derived from independent intraspecific crosses were screened, and they were subsequently found to co-segregate in hybrids derived from independent interspecific crosses here used as controls.
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  • 57
    ISSN: 1432-2242
    Keywords: Solanum ; Potato ; RAPD ; Interspecific hybrids ; SCAR
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    Topics: Biology
    Notes: Abstract A system of randomly amplified polymorphic DNA (RAPD) markers was developed to facilitate the transfer of S. bulbocastanum (blb) genes into the S. tuberosum (tbr) genome by hybridization and backcrossing. DNA from tbr, blb and the hexaploid hybrid was used as a template for polymerase chain reaction (PCR) amplification. Polymorphic RAPD products, originating from 10-mer primers, specific for blb were cloned and sequenced at their ends to allow the synthesis of 18-mer primers. The 18-mer primers allowed a more reproducible assay than the corresponding RAPDs. Of eight 18-mer primer pairs, four amplified the expected products specific for blb. However, the stringency of the primer annealing conditions needed to be carefully optimized to avoid amplification of the homeologous tbr product, suggesting that the original RAPD polymorphisms were due to single base-pair changes rather than deletions or insertions. Two primers used for amplification of backcross 2 progeny segregated in a 1∶1 (presence:absence) ratio; the other two were unexpectedly absent. The most likely explanation for the loss of these markers is irregular meiosis in the original hexaploid hybrid and subsequent elimination of chromosomes. Cytological analysis of the meiosis in the hybrid demonstrated widespread irregular pairing and the presence of lagging univalents. In addition, the first backcross individual used as the parent for the second backcross had 54 chromosomes instead of the predicted 60. In conclusion, our results demonstrate that PCR technology can be used for the efficient isolation of taxon-specific markers in Solanum. Furthermore, by the use of these markers we detected the loss of chromosomes that was subsequently shown by cytological analysis to be caused by irregular meiosis of the somatic hybrid.
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  • 58
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    Theoretical and applied genetics 91 (1995), S. 598-602 
    ISSN: 1432-2242
    Keywords: Gynogenesis ; Isozyme ; RAPD ; Agronomic evaluation ; Gametoclonal variation ; Genetic homogeneity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Haploid induction via gynogenesis offers the possibility of using doubled haploid (DH) inbred lines in onion breeding. A first DH line that originated from the open-pollinated (OP) cultivar ‘Dorata di Parma’ was obtained after overcoming difficulties associated with the haploidy of the regenerants. Spontaneous chromosome doubling occurs seldom in onion. The first DH line obtained was cloned and selfed to produce sufficient seeds for genetic studies. The homozygosity of the DH gynogenic line was revealed on the basis of the low standard deviations of the bulb traits polar diameter, shape index and weight with respect to those of the S1 line or the OP cultivar. In the DH line, moreover, segregation of RAPD and alpha esterase markers was not noted. Out of four primers revealing polymorphism at 16 ge-netic loci in the OP cultivar ‘Dorata di Parma’, none produced polymorphism in the DH gynogenic line. The Est-1 locus, homozygous in 22 plants (Est-1 1/1 in 3 and Est-1 2/2 in 19) and heterozygous (Est-1 1/2) in 11 plants of the OP cultivar, always carried the same alleles in the DH line. We also tested genetic stability during micropropagation of a second halpoid line obtained via gynogenesis from var. ‘Senshyu Yellow’. Seventeen plants of this line were tested to detect changes occurring during the tissue culture process. Again no polymorphism was observed. The high genetic homogeneity observed in the two gynogenic lines of onion could be related to the absence of the callus phase during the gynogenic process.
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  • 59
    ISSN: 1432-2242
    Keywords: Rice TGMS gene ; RAPD ; RFLP ; Molecular markers ; Gene tagging
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The thermo-sensititve genic male-sterile (TGMS) gene in rice can alter fertility in response to temperature and is useful in the two-line system of hybrid rice production. However, little is known about the TGMS gene at the molecular level. The objective of this study was to identify molecular markers tightly linked with the TGMS gene and to map the gene onto a specific rice chromosome. Bulked segregant analysis of an F2 population from 5460s (a TGMS mutant line) x ‘Hong Wan 52’ was used to identify RAPD markers linked to the rice TGMS gene. Four hundred RAPD primers were screened for polymorphisms between the parents and between two bulks representing fertile and sterile plants; of these, 4 primers produced polymorphic products. Most of the polymorphic fragments contained repetitive sequences. Only one singlecopy sequence fragment was found, a 1.2-kb fragment amplified by primer OPB-19 and subsequently named TGMS1.2. TGMS1.2 was mapped on chromosome 8 with a RIL population and confirmed by remapping with a DHL population. Segregation analysis using TGMS1.2 as a probe indicated that TGMS1.2 both consegregated and was lined with the TGMS gene in this population. It is located about 6.7 cM from the TGMS gene. As TGMS1.2 is linked to the TGMS gene, the TGMS gene must be located on chromosome 8.
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  • 60
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    Theoretical and applied genetics 91 (1995), S. 1184-1189 
    ISSN: 1432-2242
    Keywords: Nicotiana tabacum ; Chalara elegans ; Black root rot ; Resistance gene ; RAPD ; Linkage
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Linkage of randomly amplified polymorphic DNA (RAPD) markers with a single dominant gene for resistance to black root rot (Chalara elegans Nag Raj and Kendrick; Syn. Thielaviopsis basicola [Berk. and Broome] Ferraris) of tobacco (Nicotiana tabacum L.), which was transferred from N. debneyi Domin, was investigated in this study. There were 2594 repeatable RAPD fragments generated by 441 primers on DNAs of ‘Delgold’ tobacco, a BC5F8 near isogenic line (NIL) carrying the resistance gene in a ‘Delgold’ background, and ‘PB19’, the donor parent of the resistance gene. Only 7 of these primers produced eight RAPD markers polymorphic between ‘Delgold’ and ‘PB19’, indicating there are few RAPD polymorphisms between them despite relatively dissimilar pedigrees. Five of the eight RAPD markers were not polymorphic between ‘Delgold’ and the NIL. All of these markers proved to be unlinked with the resistance gene in F2 linkage tests. Of the remaining three RAPD markers polymorphic between ‘Delgold’ and the NIL, two were shown to be strongly linked with the resistance gene; one in coupling and the other in repulsion. Application of the two RAPDs in the elimination of linkage drag associated with the N. debneyi resistance gene and marker-assisted selection for the breeding of new tobacco cultivars with the resistance gene is discussed.
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  • 61
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    Theoretical and applied genetics 91 (1995), S. 1214-1221 
    ISSN: 1432-2242
    Keywords: Population genetics ; White pine blister rust ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Genetic diversity was studied in 22 populations of the white pine blister rust fungus Cronartium ribicola from natural stands and plantations of eastern white pine, Pinus strobus. Pseudo-allelic frequencies were estimated at each of 7 putative RAPD loci by scoring for presence or absence of amplified fragments in dikaryotic aecidiospores. Analysis of genetic distance between all pairs of populations did not reveal any trend with regard to geographic origin or type of white pine stand. In addition, when hierarchical population structure was analysed, total genetic diversity (H s =0.214) was mostly attributable to diversity within populations (H s =0.199; AMOVA φ st =0.121, P〈0.01). Genetic diversity of populations relative to region of origin (east, centre, and west) or type of stand (natural stands vs plantations) was not significantly different from zero (P〉0.10) Nevertheless, a significant proportion of genetic differentiation was found between populations within region or stand type (F st =0.114; φ sc =0.132, P〈0.001). This result indicates that some population structure exists but that it appears to be independent of region of origin or type of stand. At least for 2 populations from white pine plantations, it appears possible that a recent introduction of a limited number of propagules was responsible for low levels of genetic diversity. We interpret these results as meaning that either long-distance dispersal is taking place between populations more than 1000 km apart or that these populations share a common recent ancestor. In addition, we suggest that C. ribicola may still be expanding its distribution by colonizing new plantations.
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  • 62
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    Theoretical and applied genetics 98 (1999), S. 171-177 
    ISSN: 1432-2242
    Keywords: Key words Varietal identification ; RAPD ; Microsatellite ; Vitis vinifera L.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The aim of this study was to develop a cultivar identification tool based on molecular analysis and a statistical approach. From the PIC parameter we defined the D parameter, which evaluates the efficiency of a primer for the purpose of identification of varieties; i.e. the probability that two randomly chosen individuals have different patterns. D can be used to compare different types of markers even if only the allelic frequencies are known. We used this parameter to develop an algorithm for selecting the optimal combination of primers necessary to identify a set of varieties. The optimal combination of primers determined for a small elite group of varieties applied on a larger set induces a risk of confusion involving 1 of the elite varieties. We estimated the risk of confusion using the D value of each primer of the combination. We applied this methodology on a set of 224 varieties of Vitis vinifera screened with 21 RAPD primers and two microsatellite loci. The discriminating power of the primers did not only depend on the number of patterns it generates but also on the frequencies of the different patterns. A combination of 8 primers (6 RAPD and two microsatellite) was found to be optimum for the discrimination of these 224 varieties. A subset of 38 elite varieties was also investigated. The determined optimal combination of 4 primers (3 RAPD and one microsatellite) applied on the 224 varieties gave 9 risks of confusion involving 1 of the elite varieties. Confusion can happen between varieties with the same origin as well as between varieties of very diverse geographical origins.
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  • 63
    ISSN: 1432-2242
    Keywords: Key words Somatic hybridization ; Hexaploid ; RAPD ; Chromosome number variation ; Genetic improvement ; Aurantioideae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Chinese wampee [Clausena lansium (Lour.) Skeels], a sexually incompatible relative of citrus, is commercially cultivated in South China. In this study, embryogenic protoplasts of ‘Bonanza’ navel orange [Citrus sinensis (L.) Osbeck] were electrically fused with leaf protoplasts isolated from ‘Chicken Heart’ Chinese wampee. After 8 months of culture, fusion products regenerated into shoots. More than 70% of the shoots unexpectedly rooted well. Chromosome counting of several shoot- and root-tips revealed that their chromosome numbers were not 2n=4x=36 as expected, but 2n=6x=54, suggesting that chromosome doubling occurred rather than chromosome elimination in this intertribal fusion combination. RAPD analysis of embryoids and the leaves of unrooted and rooted shoots verified their hybridity. This is the first report of hexaploid somatic hybrid plant regeneration from fusion between diploids in Aurantioideae.
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  • 64
    ISSN: 1432-2242
    Keywords: Key words Vicia faba L. ; RAPD ; Mahalanobis genetic distance ; Usefulness ; Genetic variance ; Mid-parent heterosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Determining the genetic potential of a base population from the properties of their parental lines would improve the efficiency of a breeding program. In the present study, we investigated whether the means of the parents and the genetic distance determined from RAPD data (GD) or multivariate analysis (Mahalanobis D2), mid-parent heterosis (MPH), and the absolute difference between means of the parents (∣P1−P2∣) can be used for predicting the means and genetic variances (σ^2 g ) of F3:4 lines derived from different crosses in faba beans. The material comprised 18 intra- and 18 inter-pool crosses among lines from the Minor, Major, and Mediterranean germplasm pools. Fifty F3:4 lines from each cross were evaluated for days to anthesis, plant height, seeds per plant, and seed yield in German (GE) and Mediterranean (ME) environments. GD estimates between parent lines ranged from 0.38 to 0.58, while D2 ranged from 45.5 to 134.7. Correlations between means of the parents and F3:4 lines were highly significant for most traits. Estimates of σ2 g for all traits showed non-significant correlations with MPH, GD, D2. In one ME, ∣P1−P2∣ had significant associations with σ^2 g for seed yield and days to anthesis. The predicted usefulness of crosses, defined as the sum of the population mean and selection responses, was most closely associated with the means of F3:4 lines. We conclude from this study that the means of F3:4 lines can be predicted from the means of the parents, whereas the prediction of genetic variance is still an unsolved problem
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  • 65
    ISSN: 1432-2242
    Keywords: Key words Addition lines ; Multiplexed PCR ; RAPD ; Sequence tagged site ; Tritordeum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  RAPD markers were developed for octoploid×Tritordeum (amphiploid Hordeum chilense×Triticum aestivum) and its parents. Addition lines were used to identify specific RAPD markers for the Hordeum chilense chromosomes detectable in a wheat background. Twelve RAPD fragments have been cloned, sequenced and converted into STS markers. Eleven of these STSs have maintained both the chromosome specificity and the possibility of detection in a wheat background. The use of these markers in multiplexed PCRs facilitates both the efficient and reliable screening of new addition lines as well as the monitoring of introgression of H. chilense in bread and durum wheat.
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  • 66
    ISSN: 1432-2242
    Keywords: Key words Pinus contorta ; Silviculture ; Reforestation ; Gene conservation ; RAPD ; SSR ; DNA analyses
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  We examined the effects of different methods of forest regeneration on the genetic diversity of lodgepole pine (Pinus contorta var ‘latifolia’) using two different DNA-based molecular markers [randomly amplified polymorphic DNA (RAPDs) and microsatellites or simple sequence repeats (SSRs)]. Genetic diversity was estimated for 30 individuals in each of four populations for the following three stand types: (1) mature lodgepole pine (〉100 years); (2) 20- to 30-year-old harvested stands left for natural regeneration; (3) 20- to 30-year-old planted stands (4 stands of each type); and one group of 30 operationally produced seedlings. There was no significant effect of stand type on expected heterozygosity, although allelic richness and diversity were much higher for SSRs than for RAPDs. Expected heterozygosity ranged from 0.39 to 0.47 based on RAPDs and from 0.67 to 0.77 based on SSRs. The number of alleles per locus for SSRs ranged from 3 to 34 (mean 21.0), and there was a significant relationship between sequence repeat length and the number of alleles at a locus. Both marker types showed that over 94% of the variation was contained within the populations and that the naturally regenerated stands sampled had lower (not significant) expected heterozygosity than the planted or unharvested stands. The group of seedlings (assessed by RAPDs only) had expected heterozygosity and allele frequencies similar to those of the unharvested stands. Genetic distance measures were higher than obtained previously in the species using isozyme markers. There was no correlation between the two marker types for pair-wise genetic distances based on populations analyzed by both methods. Pair-wise genetic distance measures and an ordination of allele frequencies for both marker types showed little effect of geographic location or stand type on genetic similarity.
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  • 67
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    Theoretical and applied genetics 99 (1999), S. 11-15 
    ISSN: 1432-2242
    Keywords: Key words Fagus crenata ; Fagus japonica ; Microsatellite ; RAPD ; RAHM ; SSR
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  We have developed microsatellite markers (SSRs) applicable to Fagus crenata using the RAHM method and investigated their polymorphisms. We also applied the SSRs in an analysis of a closely related species, F. japonica. Here we describe the isolation and characterization of nine polymorphic microsatellite markers, of which eight are applicable to both species. Among 30 individuals of each of F. crenata and F. japonica we detected a total of 79 and 77 alleles, respectively, with an average of 9.9 and 8.6 alleles per locus. The mean expected heterozygosity (He) was 0.615 (range: 0.216–0.925) in F. crenata and 0.660 in F. japonica (range: 0.259–0.827). The He values were considerably higher than those previously found for isozymes. Paternity exclusion probabilities for multiple loci, calculated over all loci, were extremely high (0.999 and 0.998 in F. crenata and F. japonica, respectively): sufficiently high to study pollen flow in both species.
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  • 68
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    Theoretical and applied genetics 99 (1999), S. 837-843 
    ISSN: 1432-2242
    Keywords: Key words Daucus carota spp. sativus ; RAPD ; Cytoplasmic male sterility (CMS) ; Asymmetric cell fusion
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Abstract  The mitochondrial DNA of various carrot lines was characterized by random amplified polymorphic DNA (RAPD) analysis, and six sequence-tagged sites (STSs) led to identification of the petaloid type of cytoplasmic male sterility (CMS). Using six STS primer combinations, we were able to classify five CMS lines into two groups and eight fertile carrots into six groups. Both the STS1 and the STS4 primer combinations differentiated CMS cytoplasms from the fertile cytoplasms, and the STS2 primer combination revealed two different types of CMS cytoplasms – of Wisconsin Wild and Cornell origins. Cybrid carrot lines with petaloid flowers which had been obtained by asymmetric cell fusion could also be separated from fertile cybrids by the STS1 primer combination. The STS1 fragment contained a homologous sequence with the orfB gene. DNA gel blot analysis indicated that homologous regions to the STS1 fragment existed in fertile types as well as the CMS types, although the restriction fragment size patterns differed. These observations demonstrate that rearrangements involving this region occurred in the mitochondrial genome. The STS4 fragment had a more complicated gene structure, including retrotransposon-like sequences and small segments of chloroplast genome.
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  • 69
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    Theoretical and applied genetics 90 (1995), S. 189-193 
    ISSN: 1432-2242
    Keywords: Sugar beet ; Beta vulgaris ; RFLP ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract An updated map of sugar beet (Beta vulgaris L. ssp. vulgaris var ‘altissima Doell’) is presented. In this genetic map we have combined 248 RFLP and 50 RAPD loci. Including the loci for rhizomania resistance Rr1, hypocotyl colour R and the locus controlling the monogerm character M, 301 loci have now been mapped to the nine linkage groups covering 815 cM. In addition, the karyotype of some of the Beta vulgaris chromosomes has been correlated with existing RFLP and RAPD linkage maps.
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  • 70
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    Theoretical and applied genetics 90 (1995), S. 451-456 
    ISSN: 1432-2242
    Keywords: Lycopersicon esculentum ; Lycopersicon peruvianum ; RAPD ; RFLP ; Tomato spotted wiltvirus (TSWV)
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Abstract The Sw-5 locus confers dominant resistance to tomato spotted wilt virus (TSWV). To map the location and facilitate the identification of markers linked to Sw-5 we developed a pair of near-isogenic lines (NILs) and an F2 Lycopersicon esculentum x L. pennellii population segregating for resistance to TSWV. DNA from the NILs was analyzed using 748 random 10-mer oligonucleotides to discern linked molecular markers using a random amplified polymorphic DNA (RAPD) approach. One random primer (GAGCACGGGA) was found to produce a RAPD band of about 2200 bp that demonstrates linkage to Sw-5. Data from co-segregation of resistance and restriction fragment length polymorphisms (RFLPs) in a F2 interspecific population position Sw-5 between the markers CT71 and CT220 near the telomere of the long arm of chromosome 9.
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  • 71
    ISSN: 1432-2242
    Keywords: RFLP ; RAPD ; Linkage map ; Sugi (Cryptomeria japonica) ; Three-generation pedigree
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    Notes: Abstract A linkage map for sugi was constructed on the basis of restriction fragment length polymorphism (RFLP), random amplified polymorphic DNA (RAPD), and isozyme loci using a three-generation pedigree prepared for genetic analysis of heartwood color. A total of 128 RFLP (123 cDNA and 5 genomic probes), 33 RAPD, 2 isozyme, and 1 morphological (dwarf) loci segregated in 73 progeny. Of the 164 segregating loci, 145 loci were distributed in 20 linkage groups. Of these loci, 91 with confirmed map positions were assigned to 13 linkage groups, covering a total of 887.3 cM. A clustering of markers with distorted segregation was observed in 6 linkage groups. In the four clusters, distortions with a reduction in the number of homozygotes from one parent only were found.
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  • 72
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    Theoretical and applied genetics 91 (1995), S. 89-97 
    ISSN: 1432-2242
    Keywords: Garlic ; Isozyme ; RAPD ; Crop evolution ; Domestication
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    Notes: Abstract Garlic (Allium sativum L.) is a sterile species of considerable variability with respect to morphological and physiological features. The crop presumably originated in West to Middle Asia from its progenitor A. longicuspis Regel and was transported from there to the Mediterranean and other areas of cultivation. In order to clarify older classification schemes, often based on small or biased collections, we used isozyme and RAPD markers to analyze and structure a collection of 300 accessions, many of which were gathered in Middle Asia close to the assumed center of origin. All of the accessions were first investigated with isozymes, and 48 were selected for a RAPD analysis. The resulting molecular markers were used to construct neighbor-joining dendrograms to group the accessions and to indicate the genetic distances between them. Based on the dendrograms and in conjunction with some morphological features, we propose an infraspecific classification of garlic with four major groups. In agreement with the results of other workers, A. longicuspis lies within the range of the species A. sativum. Numerous forms with varying degrees of domestication are part of our longicuspis group, from which presumably the more derived cultivar groups originated. The origin and spreading of the crop are discussed with respect to the geographical distribution and the genetic distances of the accessions.
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  • 73
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    Theoretical and applied genetics 91 (1995), S. 439-447 
    ISSN: 1432-2242
    Keywords: Chrysanthemum ; RAPD ; DNA fingerprint ; RFLP ; Stability
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    Topics: Biology
    Notes: Abstract Several techniques of DNA analysis were applied to identify chrysanthemum cultivars. Unrelated cultivars could be distinguished by using RAPDs (random amplified polymorphic DNAs), inter-SSR (simple sequence repeat) PCR (polymerase chain reaction), hybridization-based DNA fingerprinting, as well as RFLPs (restriction fragment length polymorphisms). Cultivars with different flower colours and belonging to one family, i.e. vegetatively derived from 1 cultivar, appeared to have the same DNA fragment patterns, whichever technique was applied. The absence of polymorphisms between different accessions of the same cultivar indicated a high stability of the observed patterns.
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  • 74
    ISSN: 1432-2242
    Keywords: Key words Glycoalkaloids ; Potato ; Metabolic pathways ; RAPD ; Leptine ; Insect resistance ; Solanum
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    Notes: Abstract   Solanum chacoense Bitter, a wild relative of the cultivated potato, produces several glycoalkaloids, including solanine, chaconine, and the leptines. The foliar-specific leptine glycoalkaloids are believed to confer resistance to the Colorado Potato Beetle (CPB). Using two bulked DNA samples composed of high- and low-percent leptine individuals from a segregating F1 population of S. chacoense, we have identified two molecular markers that are closely linked to high percent solanine+chaconine and, conversely, to nil/low percent leptine. One of these, a 1,500-bp RAPD product (UBC370-1500), had a recombination value of 3% in the F1 progeny, indicating tight linkage. UBC370-1500 mapped to the end of the short arm of potato chromosome 1, in the region of a previously mapped major QTL for solanidine, from a S. tuberosum (solanidine)×S. berthaultii (solasodine) cross. Taken together, these results suggest that either (1) a major locus determining solanidine accumulation in Solanum spp. is on chromosome 1 in the region defined by the RFLP markers TG24, CT197, and CT233, or (2) this region of chromosome 1 may harbor two or more important genes which determine accumulation of steroidal aglycones. These findings are important for the genetics of leptine (as well as other glycoalkaloid) accumulation and for the development of CPB-resistant potato varieties.
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    Theoretical and applied genetics 98 (1999), S. 86-92 
    ISSN: 1432-2242
    Keywords: Key words Cannabis sativa ; Dioecy ; Sex ; RAPD ; SCAR
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    Notes: Abstract  A 400-bp RAPD marker generated by a primer of random decamer sequence has been found associated with the male sex phenotype in 14 dioecious cultivars and accessions of hemp (Cannabis sativa L.). The primer OPA8 generates a set of bands, most of which polymorphic among all the individual plants tested, and 1 of which, named OPA8400, present in all male plants and absent in female plants. A screening of 167 plants belonging to different genotypes for the association of the OPA8400 marker with the sex phenotype revealed that only in 3 cases was the 400-bp band was present in plants phenotypically female; on the contrary, in male plants the band was never missing, while in monoecious plants it was never present. Despite this sex-specific association, the sequences corresponding to OPA8400 were present in both staminate and carpellate plants, as revealed by Southern blotting and hybridization with the cloned RAPD band. The RAPD marker was sequenced, and specific primers were constructed. These primers generated, on the same genotypes used for RAPD analysis, a SCAR marker 390 bp in length and male-specific. This SCAR is suitable for a precise, early and rapid identification of male plants during breeding programs of dioecious and monoecious hemp.
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  • 76
    ISSN: 1432-2242
    Keywords: Key words Rosa sect. Caninae ; Biometrics ; Heterogamy ; RAPD ; Segregation distortion
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    Notes: Abstract  The dogroses, Rosa sect. Caninae, are polyploid and characterized by their unique meiosis with an unequal number of chromosomes in the male and female gametes. The pollen cells have 7 chromosomes and the egg cells 21, 28 or 35 depending on the ploidy level of the species. The resulting matroclinal inheritance was studied with both morphological and molecular markers in a pair of reciprocal crosses between R. dumalis and R. rubiginosa (2n=35). A canonical discriminant analysis based on seven morphological characters showed only a minor overlapping between the two progeny groups. In addition, the R. dumalis×R. rubiginosa offspring were more heterogeneous than the offspring from the reciprocal cross in each of the characters analysed. Eleven RAPD markers specific for the R. dumalis parent and 10 RAPD markers specific for the R. rubiginosa parent were scored in the offspring. Each of the offspring exhibited either all, or all-but-one, of the seed parent markers. The average number of pollen donor markers found in the offspring was 3.2 (R. dumalis×R. rubiginosa) and 2.7 (R. rubiginosa×R. dumalis). About half of the pollen donor markers were never transmitted to the progeny. This is, to our knowledge, the first time the highly skewed chromosome distribution in Rosa sect. Caninae has been demonstrated with statistically evaluated morphological data and with molecular markers.
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    Theoretical and applied genetics 98 (1999), S. 602-607 
    ISSN: 1432-2242
    Keywords: Key words Triticum ; Germplasm ; RAPD ; Misclassification ; Duplication
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    Notes: Abstract  Crop germplasm collections contain a considerable percentage of misclassified accessions which may affect the use of germplasm for agricultural crop improvement. The objective of this study was to determine if random amplified polymorphic DNA (RAPD) analysis could be used to reclassify misclassified Triticum accessions. Twelve accessions suspected to be misclassified, based on morphological characters, as either macha or vavilovii wheat were studied using RAPD and cytological analyses. In the RAPD analysis, a dendrogram, based on Jaccard genetic similarity coefficients, grouped 5 dicoccum-like, 1 timopheevii-like, and 6 monococcum-like accessions with Triticum dicoccum, T. timopheevii, and T. monococcum accessions, respectively. These results were confirmed by the cytological analysis. A RAPD marker specific to the D genome was also detected. This study suggests that RAPD analysis can be used to classify germplasm and to distinguish some species in Triticum.
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    Theoretical and applied genetics 98 (1999), S. 1029-1035 
    ISSN: 1432-2242
    Keywords: Key words Brassica oleracea L. ; RAPD ; Seed bulk ; Genetic resources ; Genetic variability
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Abstract  The concept of a core collection was elaborated to fit the necessity of optimizing the management, for both conservation and use, of genetic resources in sizeable collections. This approach requires an analysis of how the genetic variability is structured among the accessions. The large number of heterogeneous populations in our collection of Brassica oleracea makes genetic diversity studies based on plant-to-plant analysis impracticable. To overcome this limitation, the variability analysis by RAPD on seed bulks was investigated for its efficiency in assessing the structure of the genetic diversity of this collection. The optimal bulk size and the bulking or sampling variation were evaluated with bulks of different size and with replicated samples. A mixture of known genotypes was also used to characterise the band detection in bulks, and to compare the plant-to-plant and the bulk methods. Forty seeds were chosen to represent each population. In such a bulk, the detection of bands depended on the proportion of the genotype they were derived from in the mixture. Intense and frequent bands were detected in the bulk with a 15% detection limit. The observed bulking or sampling variation within populations was smaller than the variation between populations, leading to an efficient separation of populations with a clustering of all samples of the same population. The distances calculated from bulk data were highly correlated with the distances based on the plant-to-plant analysis. We demonstrated that RAPD on seed bulks can be used to describe the genetic diversity between populations.
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    Theoretical and applied genetics 98 (1999), S. 985-994 
    ISSN: 1432-2242
    Keywords: Key words Digitalis obscura ; AMOVA ; HOMOVA ; Population genetics ; RAPD
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Abstract  Random amplified polymorphic DNA (RAPD) markers were used to assess levels and patterns of genetic diversity in Digitalis obscura L. (Scrophulariaceae), an outcrossing cardenolide-producing medicinal plant species. A total of 50 plants from six natural populations on the Iberian Peninsula were analysed by six arbitrarily chosen decamer primers resulting in 96 highly reproducible RAPD bands. To avoid bias in parameter estimation, analyses of population genetic structure were restricted to bands (35 of 96) whose observed frequencies were less than 1–3/n in each population. The analysis of molecular variance (AMOVA) with distances among individuals corrected for the dominant nature of RAPDs (genotypic analysis) showed that most of the variation (84.8%) occurred among individuals within populations, which is expected for an outcrossing organism. Of the remaining variance, 9.7% was attributed to differences between regions, and 5.5% for differences among populations within regions. Estimates of the Wright, Weir and Cockerham and Lynch and Milligan FST from null-allele frequencies corroborated AMOVA partitioning and provided significant evidence for population differentiation in D. obscura. A non-parametric test for the homogeneity of molecular variance (HOMOVA) also showed significant differences in the amount of genetic variability present in the six populations. UPGMA cluster analyses, based on Apostol genetic distance, revealed grouping of some geographically proximate populations. Nevertheless, a Mantel test did not give a significant correlation between geographic and genetic distances. This is the first report of the partitioning of genetic variability within and between populations of D. obscura and provides important baseline data for optimising sampling strategies and for conserving the genetic resources of this medicinal species.
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  • 80
    ISSN: 1432-2242
    Keywords: Key words Cucumis melo ; Molecular markers ; RAPD ; CAPS ; RFLP ; Fusarium oxysporum ; Fusarium resistance ; Marker-assisted selection (MAS)
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Abstract  Fusarium wilt, caused by Fusarium oxysporum Schlecht f. sp. melonis Snyder & Hans, is a worldwide soil-borne disease of melon (Cucumis melo L.). Resistance to races 0 and 1 of Fusarium wilt is conditioned by the dominant gene Fom-2. To facilitate marker-assisted backcrossing with selection for Fusarium wilt resistance, we developed cleaved amplified polymorphic sequences (CAPS) and restriction fragment length polymorphisms (RFLP) markers by converting RAPD markers E07 (a 1.25-kb band) and G17 (a 1.05-kb band), respectively. The RAPD-PCR polymorphic fragments from the susceptible line ’Vedrantais’ were cloned and sequenced in order to construct primers that would amplify only the target fragment. The derived primers, E07SCAR-1/E07SCAR-2 from E07 and G17SCAR-1/G17SCAR-2 from G17, yielded a single 1.25-kb fragment (designated SCE07) and a 1.05-kb fragment (designated SCG17) (the same as RAPD markers E07 and G17), respectively, from both resistant and susceptible melon lines, thus demonstrating locus-specific associated primers. Potential CAPS markers were first revealed by comparing sequence data between fragments amplified from resistant (PI 161375) and susceptible (’Vedrantais’) lines and were then confirmed by electrophoresis of restriction endonuclease digestion products. Twelve restriction endonucleases were evaluated for their potential use as CAPS markers within the SCE07 fragment. Three (BclI, MspI, and BssSI) yielded ideal CAPS markers and were subsequently subjected to extensive testing using an additional 88 diverse melon cultigens, 93 and 119 F2 individuals from crosses of ’Vedrantais’ x PI 161375 and ’Ananas Yokneam’×MR-1 respectively, and 17 families from a backcross BC1S1 population derived from the breeding line ’MD8654’ as a resistance source. BclI- and MspI-CAPS are susceptible-linked markers, whereas the BssSI-CAPS is a resistant-linked marker. The CAPS markers that resulted from double digestion by BclI and BssSI are co-dominant. Results from BclI- and MspI-CAPS showed over 90% accuracy in the melon cultigens, and nearly 100% accuracy in the F2 individuals and BC1S1 families tested. This is the first report of PCR-based CAPS markers linked to resistance/susceptibility for Fusarium wilt in melon. The RFLP markers resulting from probing with a clone-derived 1.05-kb SCG17 PCR fragment showed 85% correct matches to the disease phenotype. Both the CAPS and RFLP markers were co-dominant, easier to score, and more accurate and consistent in predicting the melon phenotype than the RAPD markers from which they were derived.
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    Mycoscience 36 (1995), S. 465-467 
    ISSN: 1618-2545
    Keywords: benzyl bromide ; DNA preparation ; fungal DNA ; PCR ; RAPD
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    Topics: Biology
    Notes: Abstract For isolation of fungal DNA for PCR amplification, we compared three DNA isolation methods: enzymatic cleavage and the use of benzyl chloride or benzyl bromide. Since benzyl bromide is more reactive, its use enabled us to readily isolate the total nucleic acids as a DNA template source from various fungi, including dematiaceous hyphomycetes, for RAPD analysis.
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    Plant systematics and evolution 196 (1995), S. 141-151 
    ISSN: 1615-6110
    Keywords: Orchidaceae ; Loroglossum hircinum ; Compound pollen ; pollinium ; pollen tubes ; generative cell ; evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The structure of the massulae composing the pollinium ofLoroglossum hircinum was studied before pollination and 12 and 24 hours afterwards. The grains are grouped in tetrads closely packed in massulae. The exine is only present on the outside of the massulae. The intine consists of two layers: a compact layer surrounding the pollen grain and a looser layer surrounding the pollen grain and a looser layer surrounding the tetrad. Twelve hours after pollination, pollen volume and the space between the tetrads increase due to vacuolization. Twenty-four hours after pollination, pollen volume and tetrad spacing are higher due to vacuolization and some grains have emitted pollen tubes. Pollen growth due to vacuole formation, and the absence of common walls between adjacent tetrads lead to crumbling of the massulae. The mature pollen grain does not have apertures: the site of pollen tube emission is determined after pollination. The first grains to germinate are those in the centre of the massula. The vegetative cell nucleus is the first to enter the pollen tube; the generative cell elongates and undergoes the second haploid mitosis shortly after entering the pollen tube.
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    Plant systematics and evolution 198 (1995), S. 167-178 
    ISSN: 1615-6110
    Keywords: Fabaceae ; Arachis ; Arachis hypogaea ; RAPD ; systematics ; evolution ; germplasm resources
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Abstract Twenty-six accessions of wildArachis species and domesticated peanuts,A. hypogaea, introduced from South America were analyzed for random amplified polymorphic DNA (RAPD). The objective of the study was to investigate inter- and intraspecific variation and affinities among species of sect.Arachis which have been proposed as possible progenitors for the domesticated peanut. Ten primers resolved 132 DNA bands which were useful for separating species and accessions. The most variation was observed among accessions ofA. cardenasii andA. glandulifera whereas the least amount of variation was observed inA. hypogaea andA. monticola. The two tetraploid species could not be separated by using RAPDs.Arachis duranensis was most closely related to the domesticated peanut and is believed to be the donor of the A genome. The data indicated thatA. batizocoi, a species previously hypothesized to contribute the B genome toA. hypogaea, was not involved in its evolution. The investigation showed that RAPDs can be used to analyze both inter- and intraspecific variation in peanut species. Southern hybridization of RAPD probes to blots containing RAPD of theArachis species provided information on genomic relationships and revealed the repetitive nature of the amplified DNA.
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    Plant systematics and evolution 215 (1999), S. 37-47 
    ISSN: 1615-6110
    Keywords: Ranunculaceae ; Aconitum ; Delphinium ; Helleborus ; Nigella ; Seed oil ; fatty acids ; phylogeny ; evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Many members ofRanunculaceae contain unusual fatty acids in their seed oils. This leads to rather typical genus-specific fatty acid patterns or “fingerprints” in these seed oils. The members of theDelphinioideae and/orHelleboroideae, however, do not contain highly unusual fatty acids. Nevertheless, their seed oil fatty acid fingerprints are also fairly typical and genus-specific, and the patterns found are rather consistent throughout several species of one genus. It was found that species ofAconitum do not contain fatty acids with 20 carbon atoms.Delphinium, Consolida, Helleborus, Nigella and others do contain C20 fatty acids. In allHelleborus species, for example, there was a consistent C20 fatty acid pattern of 20:0≪20:1≫20:2〉20:3. Species ofNigella andGaridella contain high levels,Helleborus low levels, of 20:2n-6 in their seed oils.Delphinium andAconitum both contain low levels of 18:3n-3, whereasHelleborus spp. consistently show high levels of this fatty acid. The genus-specific fatty acid patterns found are discussed, and a correlation with the subfamily and tribe affiliation of the genera investigated here is attempted.
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  • 85
    ISSN: 1615-6110
    Keywords: Pinaceae ; Picea mariana ; P. rubens ; P. glauca ; RAPD ; genetic relationship ; interspecific hybrids ; mitotic stability
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Abstract Random amplified polymorphic DNA (RAPD) analysis was used to determine genetic relationships amongP. mariana (black spruce),P. rubens (red spruce), andP. glauca (white spruce) and to assess the degree of polymorphism within populations from different provenances and among spruce hybrids. Eleven arbitrary decamer primers were used to amplify genomic DNAs extracted from embryogenic cultures and seedlings. Species-specific RAPD markers were identified.Picea mariana andP. rubens showed similar RAPD profiles confirming their close genetic relationship. Species-specific RAPD markers were identified and were useful in distinguishing white spruce from black and red spruces. RAPD differentiation between populations within each species was small. The level of polymorphism was much higher in spruce hybrid populations than in the pure species. Cytological analysis ofP. mariana ×P. rubens hybrids showed normal mitotic behaviour at prophase, metaphase, anaphase, and telophase. All the hybrids analyzed from different cross combinations were euploids.
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    Plant systematics and evolution 216 (1999), S. 135-166 
    ISSN: 1615-6110
    Keywords: Dipsacaceae ; Pseudoscabiosa ; Scabiosa ; Sixalix ; Lomelosia ; Pycnocomon ; Scabiosiopsis ; Tremastelma ; Epicalyx ; fruit anatomy ; evolution ; systematics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Fruits ofDipsacaceae are single-seeded, have bristle-shaped calyx segments and are tightly enclosed by four fused bracts forming an epicalyx. Comparative morphological and anatomical studies reveal a great diversity of epicalyx and calyx, often relevant to fruit dispersal. The present contribution deals with theScabiosa group of genera, the core of theScabioseae tribe. Most of its taxa develop a diaphragma from a meristem on the inside of the epicalyx. This diaphragma, together with the lower part of the epicalyx encloses the fruit proper, whereas the upper parts form a so-called “epi-diaphragma” (ed) and a ± hyaline corona. Differences of the epicalyx with respect to the size and position of the ed, elaboration of the corona, origin of pits (=foveoles) and other morphological and anatomical specializations can be demonstrated. Together with palynological and karyological data these new facts support an improved concept of relationships and systematics for the taxa studied:Scabiosa sect.Scabiosa and sect.Cyrtostemma are closely related and should be united to form the genusScabiosa s. str.;Pycnocomon can be maintained as an independent genus, sister toScabiosa sect.Trochocephalus which then has to be treated as a genus,Lomelosia. In contrast, the following genera have to be included inLomelosia:Tremastelma asLomelosia sect.Callistemma, andScabiosiopsis as part ofLomelosia sect.Lomelosia. Pseudoscabiosa deviates in so many features that it has to be excluded from the redefinedScabioseae s. str.
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  • 87
    ISSN: 1615-6110
    Keywords: Gymnosperms ; Pinaceae ; A. alba ; A. cephalonica ; A. borisii-regis ; A. bornmuelleriana ; A. nordmanniana ; A. equi-trojani ; A. pinsapo ; A. numidica ; A. cilicica ; Allozyme variation ; genetic diversity ; phylogenetic relationships ; evolution
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Abstract Nineteen natural Mediterranean fir populations, belonging to eight species and to one natural hybrid (A. ×borisii-regis), were investigated by starch and polyacrylamide gel electrophoresis. A total of 31 alleles was scored at eight loci (IDH-B, ACP-A, PER-B, 6PGD-A, 6PGD-B, MNR-B, PGI-B, PGM-A. Great variation was observed in the heterozygosity among the population studied and ranged from 0.010 (A. pinsapo) to 0.328 (A. cephalonica). The interpopulation genetic diversity was about 26% of the total genetic diversity. From the dendrogram, new phylogenetic relationships were revealed. High affinity was observed between the Calabrian fir population and the one from north-west Greece as well as betweenA. equi-trojani grown in Asia Minor and the southern Greek populations. Species specific alleles were found inA. cilicica. From the findings of the present work, a new hypothesis concerning the taxonomy, distribution and evolution ofAbies species in the Balkan Peninsula is supported.
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  • 88
    ISSN: 1615-6110
    Keywords: Chlorococcales ; Microsporales ; Microspora ; Absolute configuration ; classification ; evolution ; flagellar apparatus ; ultrastructure ; zoospore
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    Topics: Biology
    Notes: Abstract The spatial configuration of the flagellar apparatus of the biflagellate zoospores of the green algal genusMicrospora is reconstructed by serial sectioning analysis using transmission electron microscopy. Along with the unequal length of the flagella, the most remarkable characteristics of the flagellar apparatus are: (1) the subapical emergence of the flagella (especially apparent with scanning electron microscopy); (2) the parallel orientation of the two basal bodies which are interconnected by a prominent one-piece distal connecting fiber; (3) the unique ultrastructure of the distal connecting fiber composed of a central tubular region which is bordered on both sides by a striated zone; (4) the different origin of the d-rootlets from their relative basal bodies; (5) the asymmetry of the papillar region which together with the subapical position of the basal bodies apparently cause the different paths of corresponding rootlets in the zoospore anterior; (6) the presence of single-membered d-rootlets and multi-membered s-rootlets resulting in a 7-1-7-1 cruciate microtubular root system which, through the different rootlet origin, does not exhibit a strict 180° rotational symmetry. It is speculated that the different basal body origin of the d-rootlets is correlated with the subapical implant of flagella. It is further hypothesized that in the course of evolution the ancestors ofMicrospora had a flagellar papilla that has migrated from a strictly apical position towards a subapical position. Simultaneously, ‘ancestral’ shift of flagella along the apical cell body periphery has taken place as can be concluded from the presence of an upper flagellum overlying a lower flagellum in the flagellar apparatus ofMicrospora. The basic features of the flagellar apparatus of theMicrospora zoospore resemble those of the coccoid green algal generaDictyochloris andBracteacoccus and also those of the flagellate green algal genusHeterochlamydomonas. This strengthens the general supposition thatMicrospora is evolutionarily closely related to taxa which were formerly classified in the traditionalChlorococcales.
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  • 89
    ISSN: 1615-6110
    Keywords: Fagaceae ; Quercus ; Hybridization ; RAPD ; allozymes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract RAPDs were employed as genetic markers to detect interspecific hybridization between the closely related oak speciesQuercus robur andQ. petraea. Fourteen primers were used in order to check the genetic status (“pure” or hybrid) of individuals classified morphologically. Among the 147 PCR fragments obtained 11 appear to be species-specific. In the phenotypically intermediate individuals different combinations of these species-specific bands were obtained. The patterns in these putative hybrids were not additive, which may be either the result of repeated backcrossing and introgression between the two species or of heterozygosity within the parental species. The results of the RAPD study are consistent with morphological analyses and allozyme data obtained for theGot-2 locus. Thus the RAPD markers used in this study may provide a powerful genetic tool for the identification of hybrids and the discrimination between the two “pure” species.
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  • 90
    ISSN: 1573-5052
    Keywords: Clonal structure ; Cloudberry ; Genetic variation ; DNA fingerprinting ; RAPD ; Rubus chamaemorus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The clonal structure of Rubus chamaemorus populations was investigated using DNA fingerprinting. The PCR-based methods included the use of 10-base RAPD primers and 16-base simple sequence repeat primers. In the hybridization method variation was studied using hypervariable multilocus probes, one derived from the M13 bacteriophage and the other a synthetic (AC)/(TG) polynucleotide. Although R. chamaemorus expresses clear variation in morphology, the level of genetic differentiation appears to be fairly low. The observed numbers of clones in the three populations examined in Finland varied from 2 to 4. The total number of genotypes across populations was 5, of which one was unique. The results obtained using the two fingerprinting methods were comparable but lead to a slightly different grouping of clones.
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  • 91
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    Journal of bioeconomics 1 (1999), S. 13-18 
    ISSN: 1573-6989
    Keywords: Malthus ; Darwin ; evolution ; policy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Economics
    Notes: Abstract This is a rather impressionist report of my recollections of the history of the bioeconomics field.
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  • 92
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    Journal of bioeconomics 1 (1999), S. 19-34 
    ISSN: 1573-6989
    Keywords: Darwinian world view ; evolution ; evolutionary economics ; development ; subjectivism ; natural selection ; analogy ; adaptation ; evolutionary progress ; preferences ; genetic endowment ; growth of consumption
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Economics
    Notes: Abstract Bioeconomics—the merging of views from biology and economics—on the one hand invites the 'export' of situational logic and sophisticated optimization developed in economics into biology. On the other hand, human economic activity and its evolution, not least over the past few centuries, may be considered an instance for fruitfully applying ideas from evolutionary biology and Darwinian theory. The latter perspective is taken in the present paper. Three different aspects are discussed in detail. First, the Darwinian revolution provides an example of a paradigm shift which contrasts most significantly with the 'subjectivist revolution' that took place at about the same time in economics. Since many of the features of the paradigmatic change that were introduced into the sciences by Darwinism may be desirable for economics as well, the question is explored whether the Darwinian revolution can be a model for introducing a new paradigm in economic theory. Second, the success of Darwinism and its view of evolution have induced economists who are interested in an evolutionary approach in economics to borrow, more or less extensively, concepts and tools from Darwinian theory. Particularly prominent are constructions based on analogies to the theory of natural selection. Because several objections to such analogy constructions can be raised, generalization rather than analogy is advocated here as a research strategy. This means to search for abstract features which all evolutionary theories have in common. Third, the question of what a Darwinian world view might mean for assessing long term economic evolution is discussed. Such a view, it is argued, can provide a point of departure for reinterpreting the hedonistic approach to economic change and development. On the basis of such an interpretation bioeconomics may not only go beyond the optimization-cum-equilibrium paradigm currently prevailing in economics. It may also mean adding substantial qualifications to the subjectivism the neoclassical economists, at the turn of the century, were proud to establish in the course of their scientific revolution.
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  • 93
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    Biology and philosophy 14 (1999), S. 395-430 
    ISSN: 1572-8404
    Keywords: adaptationism ; Daniel C. Dennett ; electric fish ; electroreception ; evolution ; evolutionary function ; indeterminism ; mental content ; neuroethology ; sensory modality ; underdetermination
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Philosophy
    Notes: Abstract Are attributions of content and function determinate, or is there no fact of the matter to be fixed? Daniel Dennett has argued in favor of indeterminacy and concludes that, in practice, content and function cannot be fixed. The discovery of an electrical modality in vertebrates offers one concrete instance where attributions of function and content are supported by a strong scientific consensus. A century ago, electroreception was unimagined, whereas today it is widely believed that many species of bony fish, amphibians, sharks, skates, and rays possess this non-human sensory modality. A look at the history of science related to this discovery reveals a highly interdisciplinary endeavor, encompassing ethology, behavioral analysis, neuroscience, and evolutionary biology. While each area provides important evidence, none is sufficient on its own to fix content and function. Instead, I argue that an interdisciplinary, neuroethological approach is required to carry out such determinations. Further, a detailed consideration of biological research suggests that while content and function claims are empirically underdetermined and uncertain, there is insufficient reason to believe in an additional problem of indeterminism. In particular, Dennett's indeterminism arises from a research methodology -- logical adaptationism -- that generates evidence from only one of the areas of neuroethology. However, logical adaptationism does not reflect adaptationism as it is practiced in contemporary biology. I conclude that Dennett is faced with a dilemma: On the one hand, he can hold to logical adaptationism and the indeterminism that results from it, while giving up the relevance of his arguments to biological practice. On the other, he can embrace a more accurate version of adaptationism -- one which plays a role in a larger neuroethological framework -- but from which no strong indeterminacy claims follow.
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  • 94
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    Biology and philosophy 14 (1999), S. 561-584 
    ISSN: 1572-8404
    Keywords: change ; evolution ; evolutionary epistemology ; selection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Philosophy
    Notes: Abstract This paper is concerned with the debate in evolutionary epistemology about the nature of the evolutionary process at work in the development of science: whether it is Darwinian or Lamarckian. It is claimed that if we are to make progress through the many arguments that have grown up around this issue, we must return to an examination of the concepts of change and evolution, and examine the basic kinds of mechanism capable of bringing evolution about. This examination results in two kinds of processes being identified, dubbed ‘direct’ and ‘indirect’, and these are claimed to exhaust all possibilities. These ideas are then applied to a selection of the debates within evolutionary epistemology. It is shown that while arguments about the pattern and rate of evolutionary change are necessarily inconclusive, those concerning the origin of novel variations and the mode of inheritance can be resolved by means of the distinctions made here. It is claimed that the process of selection in the evolution of science can also be clarified. The conclusion is that the main process producing the evolution of science is a direct or Lamarckian one although, if realism is correct, an indirect or Darwinian process plays a vital role.
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  • 95
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    Biology and philosophy 10 (1995), S. 181-196 
    ISSN: 1572-8404
    Keywords: Biological species concept ; gene flow ; gene circulation ; Ernst Mayr ; stalemates ; typology ; evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Philosophy
    Notes: Abstract Over the decades, there has been substantial empirical evidence showing that the unity of species cannot be maintained by gene flow. The biological species concept is inconclusive on this point. The suggestion is made that the unity of species is maintained rather by selection constantly spreading new alleles throughout the species, or bygene circulation. There is a lack in conceptual distinction between gene flow and gene circulation which lies at the heart of the problem. The concept of gene circulation also sheds some new light on the problem of typology and on such a broad concept as evolution. A new species definition is proposed.
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  • 96
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    Biology and philosophy 10 (1995), S. 339-356 
    ISSN: 1572-8404
    Keywords: Species ; lineage ; individual ; class ; evolution ; organism ; population
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Philosophy
    Notes: Abstract What are species? One popular answer is that species are individuals. Here I develop another approach to thinking about species, an approach based on the notion of a lineage. A lineage is a sequence of reproducing entities, individuated in terms of its components. I argue that one can conceive of species as groups of lineages, either organism lineages or population lineages. Conceiving of species as groups of lineages resolves the problems that the individual conception of species is supposed to resolve. It has added the virtue of focusing attention on the characteristic of species that is most relevant to understanding their role in evolutionary processes, namely, the lineage structure of species.
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  • 97
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    Biology and philosophy 10 (1995), S. 435-457 
    ISSN: 1572-8404
    Keywords: Functional explanation ; morphology ; ethology ; evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Philosophy
    Notes: Abstract This article deals with a type of functional explanation, viability explanation, that has been overlooked in recent philosophy of science. Viability explanations relate traits of organisms and their environments in terms of what an individual needs to survive and reproduce. I show that viability explanations are neither causal nor historical and that, therefore, they should be accounted for as a distinct type of explanation.
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  • 98
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    Biology and philosophy 10 (1995), S. 389-417 
    ISSN: 1572-8404
    Keywords: Catalysis ; chance ; determinism ; emergence of life ; evolution ; non-equilibrium thermodynamics ; panspermia ; protometabolism ; reduction ; RNA world ; self-organization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Philosophy
    Notes: Abstract This paper calls attention to a philosophical presupposition, coined here “the continuity thesis” which underlies and unites the different, often conflicting, hypotheses in the origin of life field. This presupposition, a necessary condition for any scientific investigation of the origin of life problem, has two components. First, it contends that there is no unbridgeable gap between inorganic matter and life. Second, it regards the emergence of life as a highly probable process. Examining several current origin-of-life theories. I indicate the implicit or explicit role played by the “continuity thesis” in each of them. In addition, I identify the rivals of the “thesis” within the scientific community — “the almost miracle camp.” Though adopting the anti-vitalistic aspect of the “continuity thesis”, this camp regards the emergence of life as involving highly improbable events. Since it seems that the chemistry of the prebiotic stages and of molecular self-organization processes rules out the possibility that life is the result of a “happy accident,” I claim that the “almost miracle” view implies in fact, a creationist position.
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  • 99
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    Biology and philosophy 14 (1999), S. 65-82 
    ISSN: 1572-8404
    Keywords: adaptation ; explanation ; evolution ; preadaptation ; specialization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Philosophy
    Notes: Abstract The concept of preadaptation, though useful, continues to trouble evolutionary scientists. Usually, it is treated as if it were really adaptation, prompting such diverse theorists as Gould and Vrba, and Dennett to suggest its removal from evolutionary theory altogether. In this paper, I argue that the as-if sense is ill-founded, and that the sense of preadaptation as a process may be defended as unequivocal and generally useful in evolutionary explanations, even in such problem areas as human evolution.
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  • 100
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    Biology and philosophy 14 (1999), S. 253-278 
    ISSN: 1572-8404
    Keywords: ancestry ; Bayesianism ; creationism ; Darwin ; evolution ; likelihood ; natural selection ; phylogeny ; probability ; Reichenbach
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Philosophy
    Notes: Abstract Modus Darwin is a principle of inference that licenses the conclusion that two species have a common ancestor, based on the observation that they are similar. The present paper investigates the principle's probabilistic foundations.
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