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  • Springer  (88)
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  • 1
    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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  • 2
    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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  • 3
    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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  • 4
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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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  • 5
    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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  • 6
    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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  • 7
    ISSN: 1432-1939
    Keywords: Hybridization ; Herbivores ; RAPD ; Host plant resistance ; Willow
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We studied the morphology, molecular genetics, and hebivory of two species of willows (Salix sericea and S. eriocephala) and their interspecific hybrids to test four alternative hypotheses concerning the effects of hybridization on plant resistance. Individually marked plants were identified using morphological traits in the field and measurements of stipule and leaf pubescence were made and compared using Canonical Discriminant Function Analysis. DNA was extracted from the leaves of a sample of the marked plants and RAPD-PCR analysis was performed to establish the genetic status of parental and hybrid plants. RAPD band analysis generally verified the genetic status of parental plants. Hybrid plants were usually correctly identified in the field with a few exceptions. However, the hybrid plants were a heterogeneous group of plants made up of most plants that appear to be F1s and a few plants that appear to be backcrosses to S. sericea. Morphological variables were useful for distinguishing S. sericea from S. eriocephala and hybrids, but were not as dependable in distinguishing between S. eriocephala and hybrids. We compared the densities of 11 herbivore species and the infection by a leaf rust pathogen (Melampsora sp.) on the leaves and stems of two parents and the hybrids in the field. We found support for the Additive hypothesis (3 species), the Dominance hypothesis (2 species) and the Hybrid Susceptibility hypothesis (7 species, 6 herbivores and the Melampsora rust). We found no evidence for the Hybrid Resistance hypothesis. Guild membership was not a good predictor of similar responses of species to hybrid versus parental plants. A Canonical Discriminant Function Analysis showed discrete separation of the taxa based on herbivore densities, illustrating different community structures on hybrid and parental plants. This study demonstrates the diversity of responses of phytophages in response to interspecific hybridization.
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  • 8
    ISSN: 1432-203X
    Keywords: Somatic hybrids ; Potato ; Reducing sugar ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A somatic hybridisation programme was undertaken to evaluate the expression of reducing sugar accumulation in potato. Interspecific hybrids created between the Solanum tuberosum cultivar Record and the diploid species Solanum phureja were evaluated at the morphological and molecular levels. These analyses indicated that the protoplast regenerants were partial (asymmetric) hybrids which had undergone elimination of S. phureja chromosomes. Tubers of the parents exhibited significant differences for reducing sugar accumulation during cold storage with S. phureja having lower levels of glucose and fructose than Record. The somatic hybrids resembled the S. phureja parent in terms of reducing sugar accumulation demonstrating that low reducing sugar accumulation is dominant to high reducing sugar accumulation in these particular genotypes. These results are discussed in relation to the exploitation of asymmetric hybridisation for the production of potato genotypes for the potato processing industry.
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  • 9
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    Current genetics 26 (1994), S. 456-460 
    ISSN: 1432-0983
    Keywords: Parasitella parasitica ; Zygomycetes ; RAPD ; PCR ; RFLP ; Electrophoretic karyotype ; Molecular taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In addition to conventional methods for the identification of fungi, molecular techniques at the DNA level are increasingly being employed. In order to check the validity of such experimental approaches, we have analyzed the well-defined species Parasitella parasitica, which belongs to the family Mucoraceae (Mucorales, Zygometes). The seven strains of this species, which are available from international strain collections, were analyzed by several molecular methods: restriction fragment length polymorphism analysis (RFLP), the random primer-dependent polymerase chain reaction (RAPD-PCR), and electrophoretic karyotyping. Unexpectedly, these strains are highly diverse at the molecular level. By these techniques they can be divided consistently into two different groups. Nevertheless, all seven strains belong to a single species. They show no morphological differences and sexual spores (zygospores) were found in all possible combinations either within or between the two groups. Southern-blot analysis of genomic DNA of all P. parasitica strains with RAPD-PCR-derived labelled probes shows the existence of repetitive elements characteristic for only one group of P. parasitica. In addition, chromosome sizes, which were separated by rotating-field electrophoresis, were highly divergent, and ranged from 3 to 6.5 Mb in one group and between 2 and 4.5 Mb in the other. The RAPD-PCR patterns also discriminate both groups of P. parasitica. However, they are very similar if strains of a single group are compared. Therefore, we propose that the determination of fungal species by molecular techniques should be vetted at least by morphological and physiological parameters and, whenever possible, by mating experiments.
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  • 10
    ISSN: 1432-0983
    Keywords: Aspergillus nidulans ; Aspergillus quadrilineatus ; Gene assignment ; Isoenzyme analysis ; RFLP analysis ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Chromosome-substituted haploid segregants were selected from among the benomyl-induced progeny of an interspecific hybrid produced by polyethylene-glycol-induced fusion of protoplasts of an Aspergillus nidulans ‘master strain’ and an A. quadrilineatus auxotrophic mutant. These segregants were examined by RFLP, RAPD, and isoenzyme analysis. The A. nidulans ribosomal repeat unit was assigned to chromosome V, while the benA and the pyrG genes were assigned to linkage groups VIII and I, respectively, of A. nidulans. None of the other cloned genes tested (gdhA, amdS and 25s rRNA) showed polymorphism between the two parents. The method was also used to assign RAPD markers and isoenzyme bands of β-arylesterase, phosphatases, NAD-dependent malate dehydrogenase, and cellulase, to A. nidulans chromosomes and/or to their A. quadrilineatus equivalents. The isoenzyme and DNA sequences assigned to chromosomes could be used to saturate the genetic map of A. nidulans, or could serve as starting points for the construction of a genetic map of A. quadrilineatus. No method affording the same possibilities has been described so far in Aspergilli. This chromosome-assay method may be a useful alternative to pulsed-field-gel electrophoretic procedures for the assignment of molecular markers to chromosomes.
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  • 11
    ISSN: 1432-0983
    Keywords: Phytophthora megasperma f. sp. glycinea ; Avirulence ; Gene-for-gene ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Two genetically different isolates of the homothallic Oomycete, Phytophthora sojae, were demonstrated to outcross and form hybrid oospores after co-culturing in vitro. Random amplified polymorphic DNA (RAPD) markers revealed ten hybrids among 354 oospores analysed. One F1 hybrid was allowed to self fertilise and produce an F2 population of 247 individuals. Among 53 F2 individuals, selected at random, 18 polymorphic RAPD markers were observed to segregate at near 3:1 Mendelian ratios, consistent with segregation for dominant alleles at single loci. Segregation of virulence against soybean resistance genes Rps1a, 3a, and 5 revealed that the avirulence genes Avr1a, 3a and 5 were dominant to virulence. Avirulence against these three resistance genes appeared to be conditioned by one locus for Avr1a and two independent, complementary dominant loci for both Avr3a and Avr5. Segregation of virulence against Rps6 was in the ratio of 1:2:1 (avirulent:mixed reaction:virulent), suggesting a semi-dominant allele at a single locus. Two avirulence genes and one RAPD marker formed one linkage group, in the order Avr3a, OPH4-1, Avr5, each separated by approximately 5 cM. Our results confirm that outcrossing occurred between the parental isolates, and that sexual recombination under field conditions may play an important role in generating and maintaining genetic diversity in populations of P. sojae.
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  • 12
    ISSN: 1432-0983
    Keywords: Populus tremuloides ; RAPD ; Nucleotide divergence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In the present study, a total of 142 trees sampled from five populations of trembling aspen (Populus tremuloides Michx.) in Alberta was analyzed by the polymerase chain reaction (PCR) with five random oligonucleotide primers. Null-allele frequencies of 28 putative RAPD loci were estimated using the given departure from Hardy-Weinberg equilibrium (F IS) previously estimated with isozyme markers for the same population. Nucleotide divergence between populations was then estimated in a fashion similar to restriction-fragment data, but considering the dominance of the RAPDs. The average of nucleotide divergence between populations was in the order of 0.0005 and nucleotide divergence were found to be highly correlated with geographic distance. The results suggest that isolation by distance may be an important factor in the genetic differentiation of trembling aspen.
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  • 13
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    Plant molecular biology 26 (1994), S. 597-602 
    ISSN: 1573-5028
    Keywords: apple ; DNA marker ; RAPD ; Venturia inaequalis ; Malus floribunda
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Breeding resistant apple plants is an alternative way to control fungal pathogens reducing the environmental impact due to the use of pesticides. The breeding of apple cultivars resistant to Venturia inaequalis could be much improved by marker-assisted selection. A molecular marker closely linked to the resistance locus called Vf could replace selection based on infection studies. To find such molecular markers, DNA of progenies from crossings of a resistant and a susceptible apple tree was subject to bulked segregant analysis. Two markers were found with a genetic distance of 10.6% and 19.7% recombination frequency to the Vf locus.
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  • 14
    ISSN: 1432-203X
    Keywords: Hybrid rice ; RAPD ; PCR ; Oryza sativa ; DNA fingerprinting ; Molecular markers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary DNA from three families of rice plants selected in Northern China (each comprising the male sterile, the restorer, the hybrid F1 and the maintainer lines) has been extracted and amplified by PCR with different random DNA primers (RAPD analysis). Then, DNA has been analysed by agarose gel electrophoresis and DNA bands scored as present or absent. The generated matrices are reproducible and amenable for identification of each single plant line. Thus, RAPD fingerprinting of the inbred parental lines and of the resulting hybrid is proposed as a convenient tool for the identification, protection and parentage determination of plant hybrids. Furthermore, by offering a molecular tool to verify the degree of dissimilarity between the parental lines, the RAPD analysis may also be used to search for new parental combinations.
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  • 15
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    Theoretical and applied genetics 88 (1994), S. 283-288 
    ISSN: 1432-2242
    Keywords: Picea abies ; RAPD ; Population genetics ; Linkage map
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Norway spruce (Picea abies Karst.) is a most important species among European forest trees for both economical and ecological reasons. However, this species has suffered from a lack of information on the genetic side due to the scarcity of linkage data. In this study we have used a population of 72 megagametophytes from a single tree in a natural Italian stand to produce a genetic linkage map by means of RAPD markers. Ninety-six random decamers used as primers yielded 185 polymorphic loci showing Mendelian inheritance. Analysis of the segregation by multipoint analysis allowed us to define 17 major linkage groups covering a total distance of 3584 cM, with an average spacing between markers of 22 cM. Possible uses of a genetic linkage map with respect to population ecology and genetics are discussed.
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  • 16
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    Theoretical and applied genetics 88 (1994), S. 417-422 
    ISSN: 1432-2242
    Keywords: Lotus corniculatus ; Lotus species ; Fabaceae ; interspecific hybridization ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The ability of random amplified polymorphic DNA (RAPD) to distinguish among different taxa of Lotus was evaluated for several geographically dispersed accessions of four diploid Lotus species, L. tennis Waldst. et Kit, L. alpinus Schleich., L. japonicus (Regel) Larsen, and L. uliginosus Schkuhr and for the tetraploid L. corniculatus L., in order to ascertain whether RAPD data could offer additional evidence concerning the origin of the tetraploid L. corniculatus. Clear bands and several polymorphisms were obtained for 20 primers used for each species/accession. The evolutionary pathways among the species/accessions presented in a cladogram were expressed in terms of treelengths giving the most parsimonious reconstructions. Accessions within the same species grouped closely together. It is considered that L. uliginosus which is most distantly related to L. corniculatus, may be excluded as a direct progenitor of L. corniculatus, confirming previous results from isoenzyme studies. Lotus alpinus is grouped with accessions of L. corniculatus, which differs from previous studies. With this exception, these findings are in agreement with previous experimental studies in the L. corniculatus group. The value of the RAPD data to theories on the origin of L. corniculatus is discussed.
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  • 17
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    Theoretical and applied genetics 88 (1994), S. 775-779 
    ISSN: 1432-2242
    Keywords: Beta vulgaris ; RAPD ; Mitochondrial DNA ; Chloroplast DNA ; Cytoplasmic male sterility
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The technique of random amplified polymorphic DNA (RAPD) offers a broad range of applications in the investigation of plant genomes. A promising prospect is the use of RAPD products as genetic markers. We have investigated a possible organellar source of fragments in RAPD patterns of total DNA. Two nearly-isogenic lines of cytoplasmic male-sterile and male-fertile sugar beet (Beta vulgaris L.) were subjected to RAPD analysis with six different primers. Total, nuclear, mitochondrial (mt), and chloroplast (cp), DNA from each line were investigated. Reproducible DNA fingerprints could be obtained from both organellar DNAs. Differences in band patterns of mtDNA between cytoplasmic male-sterile and -fertile lines were observed with five out of six primers, whereas different cpDNA patterns were generated by one of the primers. Consequently, the RAPD technique can be used to discriminate between different cytoplasms. Clear evidence is provided for the organellar origin of fragments in genomic (total DNA) RAPD patterns. The consequences of these results for the interpretation of RAPD analyses are discussed.
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  • 18
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    Theoretical and applied genetics 88 (1994), S. 637-645 
    ISSN: 1432-2242
    Keywords: Petunia ; RAPD ; Genetic map ; Origin Chromosome blocks
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have established the first linkage map forPetunia hybrida based upon both RAPD and phenotypical markers. The progeny studied consisted of 100 BC1 individuals derived from the [(St40xTlvl)xTlvl] back-cross. Each morphological marker has previously been mapped onto one of the seven chromosomes. The map consists of 35 RAPD loci of which 24 were affected onto chromosomes while 10 loci were not affected. The loci covered 262.9 cM with a mean distance of 8.2 cM. They are dispersed over seven linkage groups, of which six are carried on identified chromosomes. The RAPD markers were also applied on a set of tenP. hybrida, lines chosen for their diversity and on a set of seven wild species corresponding to the possible ancestors of theP. hybrida species. The markers were found both in the wild species as well as inP. hybrida lines indicating that they are inherited and are stable enough to establish similarities and to suggest relationships between species. Eight out of the ten lines carry different linkage groups of RAPD markers, which suggest that recombinant events occurred between chromosomes which originated in the wild species.
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  • 19
    ISSN: 1432-2242
    Keywords: DNA ; RFLP ; RAPD ; Brassica ; Genetic relationships
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Restriction fragment length polymorphism (RFLP) and random amplified polymorphic DNA (RAPD) markers are being used widely for evaluating genetic relationships of crop germplasm. Differences in the properties of these two markers could result in different estimates of genetic relationships among some accessions. Nuclear RFLP markers detected by genomic DNA and cDNA clones and RAPD markers were compared for evaluating genetic relationships among 18 accessions from six cultivated Brassica species and one accession from Raphanus sativus. Based on comparisons of genetic-similarity matrices and cophenetic values, RAPD markers were very similar to RFLP markers for estimating intraspecific genetic relationships; however, the two marker types gave different results for interspecific genetic relationships. The presence of amplified mitochondrial and chloroplast DNA fragments in the RAPD data set did not appear to account for differences in RAPD- and RFLP-based dendrograms. However, hybridization tests of RAPD fragments with similar molecular weights demonstrated that some fragments, scored as identical, were not homologous. In all these cases, the differences occurred at the interspecific level. Our results suggest that RAPD data may be less reliable than RFLP data when estimating genetic relationships of accessions from more than one species.
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  • 20
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    Theoretical and applied genetics 89 (1994), S. 9-13 
    ISSN: 1432-2242
    Keywords: RAPD ; Sex determination Molecular markers ; Pistachio ; Dioecy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The Random Amplified Polymorphic DNA (RAPD) technique was used to amplify DNA segments, with the objective of finding markers linked to sex determination in the dioecious species, Pistacia vera. Progenies from two female parents pollinated by a common male parent were studied. Two bulks of DNA were made in each cross, one from males and one from females, by pooling an equal weight of fresh leaves from each individual contributing to the bulk prior to DNA extraction. DNA was extracted from each bulked sample and from each of the contributing individuals. DNA was also extracted from 14 cultivars of P. vera and from 94 open-pollinated, fewweeks-old P. vera seedlings of unknown sex. Seven hundred different decamer oligonucleotide primers were used to perform DNA amplification, with 1 of these (OPO08) producing a 945 bp amplification band that was present only in the bulked female samples and absent in the bulked male samples of the two crosses. The relationship between band presence and female sex expression was conserved in every individual obtained from the two crosses and in the 14 cultivars unrelated to the crosses. We propose that this band is tightly linked to the gene(s) that control sex determination in pistachio. The OPO08945 RAPD marker could be used in a breeding program to screen the gender of pistachio plants long before they reach reproductive maturity, resulting in considerable savings of time and economic resources. In order to verify that assumption we screened 94 additional seedlings with the OPO08 primer and obtained results consistent with a 1∶1 male:female ratio.
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  • 21
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    Theoretical and applied genetics 89 (1994), S. 271-275 
    ISSN: 1432-2242
    Keywords: RAPD ; RFLP ; Chloroplast DNA ; Natural hybrid ; Paulownia taiwaniana
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Genomic DNA of Paulownia fortunei, P. kawakamii and P. taiwaniana were amplified with 10-base primers of arbitrary sequences using the polymerase chain reaction (PCR). A total of 351 DNA fragments were amplified from 23 primers and of these 265 fragments (75.5%) were polymorphic. Almost all of the PCR-amplified products of P. taiwaniana were shared by either P. fortunei or P. kawakamii, or both, and the number of polymorphic fragments shared by P. taiwaniana and P. fortunei was about equivalent to those shared by P. taiwaniana and P. kawakamii. Restriction fragments of chloroplast DNA (cpDNA) purified from Paulownia species and from reciprocal crosses between P. fortunei and P. kawakamii were analyzed. Restriction enzyme SalI-digested cpDNA showed an identical pattern in both P. kawakamii and P. taiwaniana. These results further support the hypothesis that P. taiwaniana is the natural hybrid between P. fortunei and P. kawakamii and that the maternal parent of P. taiwaniana is P. kawakamii.
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  • 22
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    Theoretical and applied genetics 89 (1994), S. 636-642 
    ISSN: 1432-2242
    Keywords: Apomixis ; Agamospermy ; Pearl Millet ; Interspecific hybrids ; RFLP ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Two molecular markers, a RAPD (randomly amplified polymorphic DNA) and a RFLP/STS (restriction fragment length polymorphism/sequence-tagged site), previously were found associated with apomictic reproductive behavior in a backcross population produced to transfer apomixis from Pennisetum squamulatum to pearl millet. The occurrence of these molecular markers in a range of 29 accessions of Pennisetum comprising 11 apomictic and 8 sexual species was investigated. Both markers were specific for apomictic species in Pennisetum. The RFLP/STS marker, UGT 197, was found to be associated with all taxa that displayed apomictic reproductive behavior except those in section Brevivalvula. Neither UGT197 nor the cloned RAPD fragment OPC-04600 hybridized with any sexually reproducing representatives of the genus. The cloned C04600 was associated with 3 of the 11 apomictic species, P. ciliare, P. massaicum, and P. squamulatum. UGT197 was more consistently associated with apomictic reproductive behavior than OPC04600 or cloned C04600, thus it could be inferred that UGT197 is more closely linked to the gene(s) for apomixis than the cloned C04600. The successful use of these probes to survey other Pennisetum species indicates that apomixis is a trait that can be followed across species by using molecular means. This technique of surveying species within a genus will be useful in determining the relative importance of newly isolated markers and may facilitate the identification of the apomixis gene(s).
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  • 23
    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
    Topics: Biology
    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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  • 24
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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
    Topics: Biology
    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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  • 25
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    Theoretical and applied genetics 87 (1994), S. 668-672 
    ISSN: 1432-2242
    Keywords: RAPD ; Genetic variation ; Upland and lowland rice ; Rice genetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Genetic variation of nine upland and four lowland rice cultivars (Oryza sativa L.) was investigated at the DNA level using the randomly amplified polymorphic DNA (RAPD) method via the polymerase chain reaction (PCR). Forty-two random primers were used to amplify DNA segments and 260 PCR products were obtained. The results of agarosegel electrophoretic analysis of these PCR products indicated that 208 (80%) were polymorphic. All 42 primers used in this experiment were amplified and typically generated one-to-four major bands. Only two primers showed no polymorphisms. In general, a higher level of polymorphism was found between japonica and indica subspecies while fewer polymorphisms were found between upland and lowland cultivars within the indica subspecies. A dendrogram that shows the genetic distances of 13 rice cultivars was constructed based on their DNA polymorphisms. Classification of rice cultivars based on the results from the RAPD analysis was identical to the previous classification based on isozyme analysis. This study demonstrated that RAPD analysis is a useful tool in determining the genetic relationships among rice cultivars.
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  • 26
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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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  • 27
    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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  • 28
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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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  • 29
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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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  • 30
    ISSN: 1432-2242
    Keywords: Key words Bs2 resistance gene ; Pepper ; RAPD ; AFLP ; Positional cloning
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    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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  • 31
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    Theoretical and applied genetics 87 (1994), S. 757-763 
    ISSN: 1432-2242
    Keywords: Lycopersicon esculentum ; RAPD ; PCR ; Meloidogyne ; Root knot nematode
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    Topics: Biology
    Notes: Abstract A PCR-based codominant marker has been developed which is tightly linked to Mi, a dominant genetic locus in tomato that confers resistance to several species of root-knot nematode. DNA from tomato lines differing in nematode resistance was screened for random amplified polymorphic DNA markers linked to Mi using decamer primers. Several markers were identified. One amplified product, REX-1, obtained using a pair of decamer primers, was present as a dominant marker in all nematode-resistant tomato lines tested. REX-1 was cloned and the DNA sequences of its ends were determined and used to develop 20-mer primers. PCR amplification with the 20-mer primers produced a single amplified band in both susceptible and resistant tomato lines. The amplified bands from susceptible and resistant lines were distinguishable after cleavage with the restriction enzyme Taq I. The linkage of REX-1 to Mi was verified in an F2 population. This marker is more tightly linked to Mi than is Aps-1, the currently-used isozyme marker, and allows screening of germplasm where the linkage between Mi and Aps-1 has been lost. Homozygous and heterozygous individuals can be distinguished and the procedure can be used for rapid, routine screening. The strategy used to obtain REX-1 is applicable to obtaining tightly-linked markers to other genetic loci. Such markers would allow rapid, concurrent screening for the segregation of several loci of interest.
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  • 32
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    Theoretical and applied genetics 87 (1994), S. 805-815 
    ISSN: 1432-2242
    Keywords: RAPD ; Bulked segregant analysis ; Genomic mapping ; Peach, Prunus persica
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Nine different F2 families of peach [Prunus persica (L.) Batsch] were analyzed for linkage relationships between 14 morphological and two isozyme loci. Linkage was detected between weeping (We) and white flower (W), 33 cM; double flower (Dl) and pillar (Br), 10 cM; and flesh color (Y) and malate dehydrogenase (Mdh1), 26 cM. A leaf variant phenotypically distinct from the previously reported wavy-leaf (Wa) mutant in peach was found in progeny of ‘Davie II’. The new willow-leaf character (designated Wa2) was closely linked (0.4 cM) to a new dwarf phenotype (designated Dw3). Two families derived from the pollen-fertile cultivar ‘White Glory’ segregated for pollen sterility, but segregation did not follow a 3∶1 ratio. Evidence is presented suggesting that ‘White Glory’ possesses a pollen-sterility gene (designated Ps2) that is non-allelic to the previously reported pollen-sterility gene (Ps) in peach. Ps2 was linked to both weeping (We-Ps2, 15.5 cM) and white flower (Ps2-W, 25.3 cM). A genomic map of peach containing 83 RAPD, one isozyme, and four morphological markers was generated using an F2 family obtained by selfing an NC174RL x ‘Pillar’ F1. A total of 83 RAPD markers were assigned to 15 linkage groups. Various RAPD markers were linked to morphological traits. Bulked segregant analysis was used to identify RAPD markers flanking the red-leaf (Gr) and Mdh1 loci in the NC174RL x ‘Pillar’ and ‘Marsun’ x ‘White Glory’ F2 families, respectively. Three markers flanking Mdh1 and ten markers flanking Gr were identified. The combination of RAPD markers and bulked segregant analysis provides an efficient method of identifying markers flanking traits of interest. Markers linked to traits that can only be scored late in development are potentially useful for marker-aided selection in trees. Alternatives for obtaining additional map order information for repulsion-phase markers in large F2 populations are proposed.
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  • 33
    ISSN: 1432-2242
    Keywords: S. halepense ; Comparative genetic mapping ; Maize ; RAPD ; Marker-assisted selection
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    Topics: Biology
    Notes: Abstract The first “complete” genetic linkage map of Sorghum section Sorghum is described, comprised of ten linkage groups putatively corresponding to the ten gametic chromosomes of S. bicolor and S. propinquum. The map includes 276 RFLP loci, predominately detected by PstI-digested S. bicolor genomic probes, segregating in 56 F2 progeny of a cross between S. bicolor and S. propinquum. Although prior cytological evidence suggests that the genomes of these species are largely homosequential, a high level of molecular divergence is evidenced by the abundant RFLP and RAPD polymorphisms, the marked deviations from Mendelian segregation in many regions of the genome, and several species-specific DNA probes. The remarkable level of DNA polymorphism between these species will facilitate development of a high-density genetic map. Further, the high level of DNA polymorphism permitted mapping of multiple loci for 21 (8.2%) DNA probes. Linkage relationships among eight (38%) of these probes suggest ancestral duplication of three genomic regions. Mapping of 13 maize genomic clones in this cross was consistent with prior results. Mapping of heterologous cDNAs from rice and oat suggests that it may be feasible to extend comparative mapping to these distantly-related species, and to ultimately generate a detailed description of chromosome rearrangements among cultivated Gramineae. Limited investigation of a small number of RFLPs showed several alleles common to S. bicolor and S. Halepense (“johnson-grass”), but few alleles common to S. propinquum and S. halepense, raising questions about the origin of S. halepense.
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  • 34
    ISSN: 1432-2242
    Keywords: Leaf rust ; RAPD ; RFLP ; Triticum aestivum ; Triticum spelta
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Near-isogenic lines (NILs) for the leaf rust resistance gene Lr9 were screened for polymorphisms at the molecular level. RAPD (random amplified polymorphic DNA) primers as well as RFLP (restriction fragment length polymorphism) markers were used. Out of 395 RAPD primers tested, three showed polymorphisms between NILs, i.e., an additional band was found in resistant lines. One of these polymorphic bands was cloned and sequenced. Specific primers were synthesized, and after amplification only resistant lines showed an amplified product. Thus, these primers define a sequence-tagged site that is specific for the translocated fragment carrying the Lr9 gene. A cross between a resistant NIL and the spelt (Triticum spelta) variety ‘Oberkulmer’ was made, and F2 plants were analyzed for genetic linkage. All three polymorphisms detected by the PCR (polymerase chain reaction) and one RFLP marker (cMWG684) showed complete linkage to the Lr9 gene in 156 and 133 plants analyzed, respectively. A second RFLP marker (PSR546) was closely linked (8±2.4 cM) to the Lr9 gene and the other four DNA markers. As this marker maps to the distal part of the long arm of chromosome 6B of wheat, Lr9 and the other DNA markers also map to the distal region of 6BL. All three PCR markers detected the Lr9 gene in independently derived breeding lines and varieties, thus proving their general applicability in wheat breeding programs.
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  • 35
    ISSN: 1432-2242
    Keywords: Brassica juncea ; RAPD ; DNA polymorphism ; Genetic distance ; Heterosis breeding
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract RAPD assays were performed, using 34 arbitrary decamer oligonucleotide primers and six combinations of two primers, to detect inherent variations and genetic relationships among 12 Indian and 11 exotic B. juncea genotypes. Of 595 amplification products identified, 500 of them were polymorphic across all genotypes. A low level of genetic variability was detected among the Indian genotypes, while considerable polymorphism was present among the exotic ones. Based on the pair-wise comparisons of amplification products the genetic similarity was calculated using Jaccard's similarity coefficients and a dendrogram was constructed using an unweighted pair group method was arithmetical averages (UPGMA). On the basis of this analysis the genotypes were clustered into two groups, A and B. Group A comprised only exotic genotypes, whereas all the Indian genotypes and four of the exotic genotypes were clustered in group B. Almost similar genotypic rankings could also be established by computing as few as 200 amplification products. In general, a high per cent of heterosis was recorded in crosses involving Indian x exotic genotypes. On the other hand, when crosses were made amongst Indian or exotic genotypes, about 80% of them exhibited negative heterosis. Results from this study indicate that, despite the lack of direct correlation between the genetic distance and the degree of heterosis, genetic diversity forms a very useful guide not only for investigating the relationships among Brassica genotypes but also in the selection of parents for heterotic hybrid combinations.
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    Theoretical and applied genetics 88 (1994), S. 412-416 
    ISSN: 1432-2242
    Keywords: Panicum ; Millet ; RAPD ; PCR ; Genetic diversity
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    Topics: Biology
    Notes: Abstract The potential use of random amplified polymorphic DNA (RAPD) was evaluated as a source of genetic markers for studying variation among four species of Panicum and within the crop species P. miliaceum and P. sumatrense. Polymorphism in RAPD markers was observed across and within species. The four species were distinct in RAPD patterns and were separated at low correlation values even with small samples involving single genotypes per species. Accessions of P. miliaceum were grouped according to geographical regions of origin. The study demonstrated that unlike isozyme and protein electrophoresis patterns, RAPD markers can be applied to studying genetic diversity, defining gene pools, and identifying cultivars for this group of millets.
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  • 37
    ISSN: 1432-2242
    Keywords: Powdery mildew (Leveillula taurica) ; Tomato ; RAPD ; RFLP ; Lv
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    Topics: Biology
    Notes: Abstract We report the tagging of a powdery mildew [Leveillula taurica (Lév.) Arnaud.] resistance gene (Lv) in tomato using RAPD and RFLP markers. DNA from a resistant (cv Laurica) and a susceptible cultivar were screened with 300 random primers that were used to amplify DNA of resistant and susceptible plants. Four primers yielded fragments that were unique to the resistant line and linked to the resistance gene in an F2 population. One of these amplified fragments, OP248, with a molecular weight of 0.7 kb, was subsequently mapped to chromosome 12, 1 cM away from CT134. Using RFLP markers located on chromosome 12, it was shown that approximately one half of chromosome 12 (about 42 cM), in the resistant variety is comprised of foreign DNA, presumably introgressed with the resistance gene from the wild species L. chilense. Further analysis of a backcross population revealed that the Lv gene lies in the 5.5-cM interval between RFLP markers, CT211 and CT219. As a prelude to map-based cloning of the Lv gene, we are currently enriching the density of markers in this region by a combination of RAPD primers and other techniques.
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  • 38
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    Theoretical and applied genetics 89 (1994), S. 590-598 
    ISSN: 1432-2242
    Keywords: Linkage map ; Brassica nigra ; RFLP ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We constructed a genetic map on Brassica nigra based on a segregating population of 83 F2 individuals. Three different types of molecular markers were used to build the map including isozymes, restriction fragment length polymorphisms (RFLP), and random amplified polymorphic DNA (RAPD). The final map contained 124 markers distributed in 11 linkage groups. The map covered a total distance of 677 cM with the markers distributed within a mean distance of 5.5cM. Of the sequences found in the B. nigra map, 40% were duplicated and organized into three different types of arrangements. They were either scattered throughout the genome, organized in tandem, or organized in blocks of duplicated loci conserved in more than 1 linkage group.
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    Theoretical and applied genetics 89 (1994), S. 964-968 
    ISSN: 1432-2242
    Keywords: RAPD ; Wheat ; Hessian fly ; DNA markers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The Hessian fly [Mayetiola destructor (Say)] is a major pest of wheat (Triticum aestivum L.) and genetic resistance has been used effectively over the past 30 years to protect wheat against serious damage by the fly. To-date, 25 Hessian fly resistance genes, designated H1 to H25, have been identified in wheat. With near-isogenic wheat lines differing for the presence of an individual Hessian fly resistance gene, in conjunction with random amplified polymorphic DNA (RAPD) analysis and denaturing gradient-gel electrophoresis (DGGE), we have identified a DNA marker associated with the H9 resistance gene. The H9 gene confers resistance against biotype L of the Hessian fly, the most virulent biotype. The RAPD marker cosegregates with resistance in a segregating F2 population, remains associated with H9 resistance in a number of different T. aestivum and T. durum L. genetic backgrounds, and is readily detected by either DGGE or DNA gel-blot hybridization.
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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
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    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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  • 41
    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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  • 42
    ISSN: 1432-2242
    Keywords: Key words Vicia faba L. ; RAPD ; Mahalanobis genetic distance ; Usefulness ; Genetic variance ; Mid-parent heterosis
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    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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  • 43
    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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  • 44
    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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  • 45
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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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  • 46
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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
    Topics: Biology
    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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  • 47
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    Theoretical and applied genetics 89 (1994), S. 847-852 
    ISSN: 1432-2242
    Keywords: RAPD ; DNA fingerprinting ; Phylogeny Apple rootstocks ; Identification key
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract RAPD (random amplified polymorphic DNA) markers were used to fingerprint eight commercially available apple rootstocks (Nertchinsk, Northern Spy, Osman, Heyer 12, M.1, M.9, M.26 and MM.106), 10 winter hardy offsprings derived from the cross of Nertchinsk x M.9, six winter hardy offsprings derived from the cross of Nertchinsk x M.26 and one winter hardy offspring derived from each of the two crosses between Osman x Heyer 12 and Northern Spy x M.1. Phylogeny analysis using parsimony allowed us to draw the genetic relationship between these lines using only RAPD markers data. The resulting cladogram was compared to the true genetic relationship between these lines in order to assess the efficiency of RAPD markers in determining accurately the phylogenetic relationship. We also developed a DNA fingerprinting system based on 13 informative RAPD loci amplified by five RAPD primers that allowed the rapid identification of apple rootstocks.
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  • 48
    ISSN: 1432-2242
    Keywords: Key words Glycoalkaloids ; Potato ; Metabolic pathways ; RAPD ; Leptine ; Insect resistance ; Solanum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    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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  • 49
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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
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    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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  • 50
    ISSN: 1432-2242
    Keywords: Key words Rosa sect. Caninae ; Biometrics ; Heterogamy ; RAPD ; Segregation distortion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    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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  • 51
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    Theoretical and applied genetics 98 (1999), S. 602-607 
    ISSN: 1432-2242
    Keywords: Key words Triticum ; Germplasm ; RAPD ; Misclassification ; Duplication
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    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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  • 52
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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
    Topics: Biology
    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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  • 53
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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
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    Topics: Biology
    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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  • 54
    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
    Topics: Biology
    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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  • 55
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    Plant systematics and evolution 189 (1994), S. 133-148 
    ISSN: 1615-6110
    Keywords: Fabaceae ; Vicia pisiformis ; rDNA ; rflp ; PCR ; RAPD ; random priming ; genetic variation ; rare species ; threatened species
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    Topics: Biology
    Notes: Abstract Nine Swedish populations, 1–5 individuals/population, and one cultivated individual of the rare speciesVicia pisiformis were investigated for genetic variation. In hybridizations with two rDNA probes using 8 restriction enzymes, only two individuals belonging to one population were polymorphic. A map of the rDNA gene cluster was constructed for four of the restriction enzymes used. Two of the polymorphic sites were mapped and were found to be located outside regions coding for rRNA, presumably caused by single point mutations or small deletions. The repeat length of the rDNA region was c. 10,000 bp, which corresponds well with the size found for other species belonging toFabaceae. No length polymorphism was found in the intergenic spacer, contrary to the situation found for most other plant species investigated for rDNA variation. The haplotype diversity for the species (Hsp Shannon) was very low (0.055). Within-population values (Hpop) was 0 for all populations except the variable one, which had 0.301. PCR amplification with 6 random primers also revealed very low levels of genetic diversity. A polymorphism was observed in a limited number of individuals for four populations. Hsp was 0.065 and $$\bar Hpop$$ was 0.050. The average D value (Wetton) for the PCR haplotypes was 0.99.
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  • 56
    ISSN: 1615-6110
    Keywords: Pinaceae ; Picea mariana ; P. rubens ; P. glauca ; RAPD ; genetic relationship ; interspecific hybrids ; mitotic stability
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    Topics: Biology
    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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  • 57
    ISSN: 1615-6110
    Keywords: Fagaceae ; Quercus ; Hybridization ; RAPD ; allozymes
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    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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  • 58
    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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  • 59
    ISSN: 1572-9788
    Keywords: Musa ; plantain ; RAPD ; VNTR ; AFLP ; breeding
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Progress in the breeding of plantain and banana has been restricted by the complex genetic structure and behaviour of cultivated polyploid Musa. Genetic improvement has been hindered due to the large amount of space required for growth and maintenance of plant populations, in addition to the long growth cycle and the low levels of fertility and seed viability characteristic of cultivated genotypes. Molecular marker assisted breeding has the potential to dramatically enhance the pace and efficiency of genetic improvement in Musa. This study was conducted to compare different PCR-based marker systems (RAPD, VNTR and AFLP) for the analysis of breeding populations generated from two diverse Musa breeding schemes. All three assays detected a high level of polymorphism between parental genotypes and within progeny populations. As expected, AFLP assays had by far the highest multiplex ratio while VNTR analysis detected the highest levels of polymorphism. AFLP analysis of a full-sib tetraploid hybrid population confirmed previous reports based on VNTR analysis, of a high frequency of recombination during 2n (3x) gamete formation by a triploid plantain landrace. In addition, both VNTR and RAPD analyses of a full-sib triploid hybrid population suggested a high frequency of homoeologous recombination during n (2x) gamete formation by tetraploid hybrids. In general, there was a poor correlation between estimates of genetic similarity based on different types of marker. The implications of these findings for the molecular breeding of Musa crops are discussed.
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  • 60
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    Molecular breeding 5 (1999), S. 275-281 
    ISSN: 1572-9788
    Keywords: AFLP ; DNA markers ; genetic mapping ; marker-aided selection ; Pinus radiata ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Efficient construction of reasonable density genetic linkage maps is an essential component of QTL detection programmes. The AFLP technique has been used to produce genetic linkage maps in a range of species. We have developed protocols to generate reproducible AFLP profiles in Pinus radiata and have evaluated the inheritance and informativeness of AFLP markers in this important timber species. The large genome size of P. radiata necessitated increased levels of selection at both the pre-amplification and selective amplification steps of the AFLP protocol to generate reproducible AFLP profiles. Once optimised ca. 41.3 scorable AFLP bands were resolvable through denaturing gels, of which 48.4% were polymorphic in a screen of eight unrelated trees. This level of polymorphism is ca. three times higher than with RAPD markers. The total number of bands and the number of polymorphismic bands per PCR were ca. halved when AFLPs were electrophoresed on non-denaturing gels and stained with ethidium bromide. Using the protocols developed, AFLP is an efficient method for generating the DNA markers required for genetic linkage map construction in P. radiata.
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  • 61
    ISSN: 1573-8590
    Keywords: Artemia ; genetic polymorphism ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geography
    Notes: Abstract We have applied the technique of random amplification of polymorphic DNA (RAPD) to the analysis of the relationships among four species of brine shrimp:Artemia franciscana, A. urmiana, A. sinica, andA. parthenogenetica. Seventy ten-base synthetic oligonucleotides were used to amplify a total of 458 distinct fragments. DNA polymorphisms were found in all the species examined; the highest percentage of polymorphic bands was found inA. parthenogenetica, with 28.8 per cent. Each species was scored for the presence or absence of every amplification product and the data entered into a binary data matrix. Cluster analysis was then performed to create a dendrogram using UPGMA by the NTSYS program. There are significant differences between bisexual species and parthenogenetic populations.A. parthenogenetica provided 94 specific molecular markers, while bisexual species gave 27 specific molecular markers.A. sinica is a species distinct from the other Old World bisexual species.
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  • 62
    ISSN: 1573-8590
    Keywords: Artemia ; genetic polymorphism ; RAPD
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    Topics: Biology , Geography
    Notes: Abstract We have applied the technique of random amplification of polymorphic DNA (RAPD) to the analysis of the relationships among four species of brine shrimp: Artemia franciscana, A. urmiana, A. sinica, and A. parthenogenetica. Seventy ten-base synthetic oligonucleotides were used to amplify a total of 458 distinct fragments. DNA polymorphisms were found in all the species examined; the highest percentage of polymorphic bands was found in A. parthenogenetica, with 28.8 per cent. Each species was scored for the presence or absence of every amplification product and the data entered into a binary data matrix. Cluster analysis was then performed to create a dendrogram using UPGMA by the NTSYS program. There are significant differences between bisexual species and parthenogenetic populations. A. parthenogenetica provided 94 specific molecular markers, while bisexual species gave 27 specific molecular markers. A. sinica is a species distinct from the other Old World bisexual species.
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    European journal of plant pathology 105 (1999), S. 667-680 
    ISSN: 1573-8469
    Keywords: asexual reproduction ; mating types ; oomycetes ; origin ; RAPD ; RFLP ; population genetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Phytophthora cinnamomi isolates from South Africa and Australia were compared to assess genetic differentiation between the two populations. These two populations were analysed for levels of phenotypic diversity using random amplified polymorphic DNAs (RAPDs) and gene and genotypic diversity using restriction fragment length polymorphisms (RFLPs). Sixteen RAPD markers from four decanucleotide Operon primers and 34 RFLP alleles from 15 putative loci were used. A few isolates from Papua New Guinea known to posses alleles different from Australian isolates were also included for comparative purposes. South African and Australian P. cinnamomi populations were almost identical with an extremely low level of genetic distance between them (Dm=0.003). Common features for the two populations include shared alleles, low levels of phenotypic/genotypic diversity, high clonality, and low observed and expected levels of heterozygosity. Furthermore, relatively high levels of genetic differentiation between mating type populations (Dm South Africa=0.020 and Dm Australia=0.025 respectively), negative fixation indices, and significant deviations from Hardy–Weinberg equilibrium, all provided evidence for the lack of frequent sexual reproduction in both populations. The data strongly suggest that both the South African and Australian P. cinnamomi populations are introduced.
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  • 64
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    Plant systematics and evolution 217 (1999), S. 313-332 
    ISSN: 1615-6110
    Keywords: Cucumis melo ; melon ; intra-specific classification ; RAPD ; Inter-SSR ; DNA fingerprinting
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cucumis melo L. (melon) genotypes differ widely in morphological and biochemical traits. Intraspecific classification of such variability has been difficult, and most taxonomists still rely on the work of Naudin (1859). A collection of 54 accessions representing diverse genotypes from 23 countries was surveyed. Morphological traits related to the vegetative and flowering stages and mature fruit morphology and quality parameters, e.g., taste, aroma, sugar composition and pH, were scored. These were used to construct a “botanical-morphological” dendrogram that generally reflected the classification ofCucumis melo into several horticultural varieties. DNA polymorphism among the accessions was assessed using the Inter-SSR-PCR and RAPD techniques that detected abundant DNA polymorphism among melon genotypes. Cluster analysis indicated that the largest divergence was between North American and Europeancantalupensis andinodorus cultivars as one group, and the more “exotic” varieties:conomon, chito, dudaim, agrestis andmomordica, as a second group. The molecular phylogeny agreed, broadly, with the classification of melon into two subspecies, and did not contradict the division into “horticultural varieties”. It was apparent, however, that the infra-specific division is rather loose, molecular variation being distributed continuously between and within cultivar groups. We suggest that despite the morphological diversity, separation between varietal-groups may be based on a too small number of genes to enable unambiguous infra-specific classification based on DNA diversity.
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    Plant and soil 159 (1994), S. 159-170 
    ISSN: 1573-5036
    Keywords: DNA-mediated transformation ; DNA polymorphisms ; ectomycorrhizal fungi ; genetics ; polymerase chain reaction ; RAPD ; symbiosis-related genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The variability within and among ectomycorrhizal species provides a substantial genetic resource and the potential to increase forest productivity and environmental sustainability. Two parallel and interacting approaches, classical and molecular genetics, are being developed to acquire the genetic information underpinning selection of improved ectomycorrhizal strains. Determining the genetic traits of the fungi which contribute to symbiosis and plant function are being followed using natural variability combined with classical and molecular genetic manipulations. Classical and molecular manipulations for breeding rely on key information including sexual and parasexual reproduction, postmeiotic nuclear behaviour, mating-types and vegetative incompatibility mechanisms. Progress in the manipulation of genomes of ectomycorrhizal fungi will depend on efficient methods for gene cloning and DNA transformation. Gene transfer into fungal cells have been shown to be successful and include treatment of protoplasts and intact mycelium with naked DNA in the presence of polyvalent cations, electroporation, and microbombardment. The merits and limitations of these methods are discussed. Using this technology the expression of foreign DNA, the functional analysis of fungal DNA sequences, as well as molecular exploitation for commercial purposes can be carried out. This review concentrates on these aspects of fungal molecular biology and discusses the applications of the experimental systems that are currently available to ectomycorrhizal fungi. As it is essential to be able to define the traits which a breeder is seeking to improve, availability of genetically defined strains that are isogenic for a character or differ only in one character and a thorough knowledge of the biochemistry of the symbiosis will be necessary before any genetic manipulation be carried out. Genetic variability of ectomycorrhizal strains has been assessed by DNA fingerprinting. This approach allows the evaluation of DNA variability and the exchange of genetic information in natural populations, the identification of species and isolates by DNA polymorphisms, and tracking the environmental fate of the introduced fungi to determine their survival, growth, and dissemination within the soil.
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  • 66
    ISSN: 1573-5060
    Keywords: Brassica napur ; doubled haploids ; RAPD ; linolenic acid ; erucic acid ; marker assisted selection ; rapeseed breeding
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Undesirable characteristic of rapeseed oil is a relatively high level of linolenic acid (18:3), which is easily oxidized leading to rancidity and a shortened shelf life of the oil. Previous attempts to reduce linolenic acid levels in rapeseed oil through breeding have been impaired by complex genetics and strong environmental sensitivity of this trait. Therefore, our objective was to develop molecular markers for low linolenic acid that could facilitate the breeding of low linolenic rapeseed. Bulked segregant analysis was employed to identify two RAPD markers associated with 18:3 in a doubled haploid population segregating for linolenic and erucic acid levels. Based on analysis of individual DH lines, the markers RM350 and RM574, representing two independent loci, accounted for a total of 39% of the genetic variability in this population. This marker RM350 alone accounted for 25% genetic variation for this trait with no evidence of recombination. Significant interlocus interaction found between the markers RM350 and RM574 suggested that epistasis was involved in the genetic control of 18:3 level in this population. Another marker designated as RM322, which was independent of the other two, was found significantly associated with the erucic acid level and oil content. RAPD markers identified in this study should be a useful tool for the early detection of low linolenic, or low or high erucic acid genotypes in rapeseed breeding programs based on doubled haploids.
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  • 67
    ISSN: 1573-5060
    Keywords: crown rot ; fusarium crown and root rot ; genetic linkage ; Lycopersicon peruvianum ; molecular markers ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract In tomato ( Lycopersicon esculentum Mill.) a single dominant gene ( Frl) on chromosome 9 confers resistance to fusarium crown and root rot (crown rot) incited by Fusarium oxysporum f. sp. radicis-lycopersici. To identify randomly amplified polymorphic DNA (RAPD) markers linked to Frl, crown rot susceptible and resistant tomato lines were screened for polymorphisms using 1000 random 10-mer primers and three reliable RAPD markers were found linked to Frl (UBC #'s 116, 194, and 655). A codominant polymorphic PCR marker of TG101, a restriction fragment length polymorphic (RFLP) marker linked to Frl, was developed to facilitate the linkage studies. Using TG101 and the four RAPD markers, on a Frl segregating backcross population of 950 plants indicated that all belong to the same linkage group. The polymorphic allele order was found to be TG101 – 655 – 116 – 194 – Frl. UBC 194 was found to be 5.1 cM from Frl in this population. Furthermore, it was the only marker found in the resistant genotypes ‘Mocis’ and Fla 7226, whereas resistant genotypes ‘Momor’, Ohio 89-1, and Fla 7464 all had UBC 194 and UBC #'s 116, 194, and 655.
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  • 68
    ISSN: 1573-5060
    Keywords: Bph-1 ; linkage analysis ; mapping ; RAPD ; RFLP ; rice
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract We report the tagging of a brown planthopper (BPH) resistance gene (Bph–1) in rice using RAPD and RFLP markers. The Korean rice variety ‘Gayabyeo’ has dominant duplicate genes including Bph–1 conferring resistance to biotype 1 of BPH. Bulked segregant RAPD analysis was employed for rapid identification of DNA markers linked to resistance genes. For tagging these two genes, an F2F3 population from a ‘Gayabyeo’ × ‘Nagdongbyeo’ cross was developed and evaluated for BPH resistance. Three bulked DNAs from two groups of homozygous BPH resistant (each for Bph–1 and the other unknown gene) and homozygous susceptible F2 plants were analyzed by RAPD using 140 random oligomers. One primer, OPD–7 yielded a 700-bp fragment that was present in Gayabyeo and resistant F2 plants (homozygous for Bph-1 locus) but absent in Nagdongbyeo and susceptible F2 plants. Cosegregation of this marker with Bph-1 was verified using an F2 population segregating for Bph-1. Chromosomal regions surrounding the Bph-1 were examined with additional RFLP and microsatellite markers on chromosome 12 to define the location of the RAPD marker and Bph-1. Use of this RAPD marker could facilitate early selection of resistant lines for BPH.
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  • 69
    ISSN: 1573-5060
    Keywords: AFLP ; DNA fingerprinting ; isozymes ; RAPD ; rice
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A survey of the genetic diversity among the major cuban rice cultivars was conducted using isozyme, RAPD and AFLP markers. Polymorphisms were detected for esterases, peroxidases, alcohol dehydrogenases and polyphenoloxidases systems; 21 RAPD primers and four AFLP primer combinations. Heterozygosity arithmetic mean value (Hav(p)), the effective multiplex ratio (EMR) and the marker index (MI), were calculated for isozyme, RAPD and AFLP markers. The mean value of genetic similarity among the different varieties was 0.92 for isozyme, 0.73 for RAPD and 0.58 for AFLP analyses. Thus, AFLP were able to detect polymorphisms with higher efficiency than RAPD (+15%) and isozyme (+34%). Data from the isozyme, RAPD and AFLP analyses were used to compute matrices of genetic similarities. The efficiency of the UPGMA for the estimation of genetic relatedness among varieties was supported by cophenetic correlation coefficients. The resulting values indicated that the distortion level for the estimated similarities was minimal. The correlation coefficients obtained by the Mantel matrix correspondence test, which was used to compare the cophenetic matrices for the different markers, showed that estimated values of genetic relationship given for isozyme and RAPD markers (r = 0.89), as well as for AFLP and RAPD markers (r = 0.82) were properly related. However, AFLP and isozyme data showed only moderate correlation (r = 0.63). Although the genetic variability found among the different cultivars was low, both RAPD and AFLP markers proved to be efficient tools in assessing the genetic diversity of rice genotypes.
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  • 70
    ISSN: 1573-5060
    Keywords: classification ; DNA ; plum varieties ; Prunus ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract We investigated the genetic diversity of 42 plum varieties by RAPD analysis. Twenty primers discriminated all plum varieties excepting two synonymous pairs: 'Botankyou and ‘Kelsey’, and ‘Chairn’ and ‘Tragedy’. Two North American plums, ‘Beach Plum’ and ‘Glow’, were genetically distinct from the other plums by cluster analysis. Overlaps observed between the ‘European plum group’ and the ‘Japanese plum group’, were perhaps due to intercrossing. We could also discriminate ‘Sordum’ from 'Late Sordum and ‘Bansei Sordum’, although ‘Late Sordum’ and ‘Bansei Sordum’ are thought to be derived from bud mutants of ‘Sordum’.
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  • 71
    ISSN: 1573-5060
    Keywords: DNA fingerprint ; Lactuca sativa ; Oryza sativa ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Thirty-five rice (Oryza sativa L.) varieties, including 18 japonica, 5 javanica and 12 indica subspecies and 12 lettuce (Lactuca sativa L.) varieties were identified taxonomically, using PCR with originally designed 21 RAPD (Random Amplified Polymorphic DNA) primers and 8 sequence-specific primers, used for amplifying four specific DNA fragments. Use of these primers revealed polymorphisms among varieties in rice and lettuce and facilitates DNA fingerprinting. Dendrograms of both species based on polymorphisms were constructed and genetical relationships were established. In rice, half the number of amplified bands were polymorphic and almost all varieties differentiated. However, differentiation of minor genetic alterations among somaclonal variants or mutants and their mother varieties was not feasible. In L. sativa, 47% of the amplified fragments were polymorphic and all 12 varieties were differentiated. Some of the PCR fragments were variety or type specific, which could be used for indicators for type-selection. The dendrogram obtained showed differentiated clusters of crisphead, leaf and butterhead type, findings in good accord with the classification based on the genetic background.
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  • 72
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    Genetic resources and crop evolution 46 (1999), S. 587-598 
    ISSN: 1573-5109
    Keywords: diversity ; genetic resources ; GIS ; Oryza sativa ; RAPD ; rice
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A diverse set of 115 rice varieties from Bangladesh was surveyed using 35 polymorphic RAPD (randomly amplified polymorphic DNA) markers and the genetic structure of this germplasm, encompassing the principal rice ecotypes of Bangladesh (aus, aman and boro), was determined using multivariate analysis. The level of genetic diversity was evaluated and compared with the levels of diversity found within other rice growing areas of the world. Geographical information systems analysis using Atlas-GIS was employed to analyse and present the geographic distribution of genetic diversity across Bangladesh, and cluster analysis was used to test the efficiency of selection of material for a core collection.
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  • 73
    ISSN: 1573-5109
    Keywords: core collection ; germplasm ; molecular marker ; potato ; RAPD ; Solanum phureja
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The potato crop originated in the Andean highlands where numerous farmer's varieties and non-cultivated wild species exist. An Andean potato collection is held in trust at the International Potato Center (CIP) to preserve the biodiversity of this crop and ensure the supply of germplasm for potato improvement worldwide. A core collection representing the biodiversity of the Andean potato germplasm is under construction using morphological, molecular, and geographic data. One of the eight cultivated potato species, Solanum phureja, has been genotyped using the RAPD technique. A protocol suitable for large germplasm collection genotyping has been developed to process numerous samples at reasonable costs. From 106 RAPD primers evaluated, we have selected 12 primers yielding 102 polymorphic markers, which unambiguously discriminated all 128 accessions but 2 that are possible duplicates. The S. phureja germplasm collected throughout the Andean countries appears to have a homogeneous genetic constitution. There was no clear geographic pattern as indicated by cluster analysis of the RAPD data. A sub-group of 20 accessions has been identified on the basis of the marker data and selected to maximize molecular (RAPD) variance and polymorphism. The probability of capturing equal amounts of marker polymorphism in this sub-group of 20 accessions by random sampling is less than 40%. This set accessions represents our first group of accessions that may constitute a core of the S. phureja collection. This tentative core will be challenged for diversity content by alternate markers and agronomic traits. Hence, the methodology for sampling less than 10% of the base collection, proposed for core collections by Brown (1989), can be based on molecular marker data provided cost-efficient fingerprints are developed.
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  • 74
    ISSN: 1573-6857
    Keywords: barley ; Hordeum spontaneum ; microsite ecology ; molecular edaphicdifferentiation ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Random amplified polymorphic DNA polymerase chain reaction (RAPDPCR) was used to assess genetic diversity in four subpopulations (86 individuals) of wild barley, Hordeum spontaneum, sampled from Tabigha microsite near the Sea of Galilee, Israel. The microsite consists of two 100 m transects that are topographically separated by 100 m, each equally subdivided into 50 m of basalt and terra rossa soil types. Despite the same macroclimate characterizing the area around the Sea of Galilee, the microsite offers two edaphically different microhabitats, with basalt being a more ecologically heterogeneous and broader-niche than the relatively drier but more homogeneous and narrow-niche terra rossa. Analysis of 118 putative loci revealed significant (P〈0.05) genetic differentiation in polymorphism (P0.05) between the two soils across the transects with P being higher in the more heterogeneous basalt (mean P0.05 = 0.902), than in terra rossa (mean P0.05 = 0.820). Gene diversity (He) was higher in basalt (mean He=0.371), than in terra rossa (mean He=0.259). Furthermore, unique alleles were confined to one soil type, either in one or both transects. Rare alleles were observed more frequently in terra rossa than basalt, and in transect II only. Gametic phase disequilibria showed a larger multilocus association of alleles in basalt than terra rossa, and in transect I than II. Spearman rank correlation (rs) revealed a strong association between specific loci and soil types, and transects. Also, analysis of multilocus organization revealed soil-specific multilocus-genotypes. Therefore, our results suggest an edaphically differentiated genetic structure, which corroborates the niche width-variation hypothesis, and can be explained, in part, by natural selection. This pattern of RAPD diversity is in agreement with allozyme and hordein protein diversities in the same subpopulations studied previously.
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  • 75
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    Plant cell, tissue and organ culture 59 (1999), S. 81-87 
    ISSN: 1573-5044
    Keywords: asymmetric hybrid ; Citrus ; donor-recipient fusion ; Microcitrus ; RAPD ; X-ray
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract X-ray irradiated embryogenic protoplasts of Microcitrus papuana Swing. were electrically fused with iodoacetic acid-treated embryogenic protoplasts of Newhall navel orange [Citrus sinensis (L.) Osb.]. Seven cell lines were established by low-melting agarose embedding culture of fusion-treated protoplasts. Cytological examination of 4 cell lines showed that each cell line consisted of many aneuploid (45.10%, 38.98%, 32.69% and 34.85%, respectively) and diploid cells (52.94%, 59.33%, 63.46% and 62.12%. respectively), whereas only a few tetraploid cells (1.96%, 1.69%, 3.85% and 3.03%, respectively) were detected. Analyses of random amplified polymorphic DNA with four 10-mer primers confirmed the hybrid characteristics of the cell lines, which in combination with chromosome counting proved that the cell lines were asymmetric hybrids.
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  • 76
    ISSN: 1573-5060
    Keywords: Erwinia carotovora ; Solanum tuberosum ; somaclonal variation ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Three somatic hybrid lines between potato (cv. While Lady line no. Ke 79, 2n = 2x = 48) + Solanum brevidens (PI 218228, 2n = 2x = 24) were evaluated using randomly amplified polymorphic DNA (RAPD) markers. The lines originated from the same callus but showed different reactions to Erwinia carotovora ssp. carotovora, the cause of potato soft rot. By the use of 48 oligomer primers producing 99 scorable bands, DNA polymorphism were detected on 7 of 12 S. brevidens chromosomes. Loss of certain DNA segments on chromosome 5, 6, 9 and 11 were observed. Some of the variations could have taken place in early callus stage of development; others may have occurred after initiation of individual shoot regeneration. The possible involvement of missing RAPD products specific to one somatic hybrid that shows decreased resistance to bacterial soft rot is discussed.
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  • 77
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    Euphytica 107 (1999), S. 167-176 
    ISSN: 1573-5060
    Keywords: genetic diversity ; Lathyrus ; L.sativus ; phylogenetic relationship ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Eight Lathyrus sativus L. accessions from a variety of geographic origins were used to study intraspecific genetic diversity using RAPD analysis. Fourteen decamer primers produced 64 amplification products, 50% of which were polymorphic between the samples. Jaccard's coefficient of genetic similarity was calculated between samples and a dendrogram was constructed by an unweighted pair-group method with arithmetical averages (UPGMA). The dendrogram showed that most of the L. sativus plants clustered into accessions or common geographical areas. The average genetic similarity coefficient within accessions was 0.12 and between accessions was 0.20, indicating a low level of intraspecific genetic variation. Interspecific genetic diversity and phylogenetic relationships of eight Lathyrus species, including L. sativus and Pisum sativum L. (field pea) were examined using 14 decamer primers which produced 283 amplification products. All amplification products were polymorphic across the nine species. In the dendrogram the Lathyrus species clustered into three distinct groups which correlated with the Sections Lathyrus, Clymenum and Linearicarpus. This supports traditional taxonomic classifications of the genus Lathyrus which are based on morphological traits. Of the species from Section Lathyrus, L. gorgoni and L. cicera were the most similar to L. sativus. The results suggest that a strategy of breeding for producing lines of L. sativus with increased genetic variation would be effectively achieved through hybrid production between accessions from wide geographic areas particularly the Mediterranean area and the Indian subcontinent. However, the most effective method would be introgression of germplasm from other species in Section Lathyrus.
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  • 78
    ISSN: 1573-5060
    Keywords: genetic diversity ; germplasm ; molecular markers ; RAPD ; sesame
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Fifty-eight accessions of sesame (Sesamum indicum L.), an important oil seed crop of the tropics and subtropics were analysed using random amplified polymorphic DNA (RAPD) technique. The material analysed comprised 36 collections from 18 different states of India and four adjoining countries of the Indian subcontinent, and 22 exotic accessions from 21 sesame growing countries around the world. The results from PCR amplifications with the selected 24 random 10-mer primers were statistically analysed. The value of Jaccard’s similarity coefficients ranged from 0.19 to 0.89. The results indicated the presence of high level of genetic diversity. However, the extent of genetic diversity was greater in the collections from Indian subcontinent as compared to the exotics. Among the Indian accessions, the collections from Rajasthan and North-eastern states were highly diverse. The phenetic analysis grouped 48 out of 58 accessions in six clusters and the remaining highly diverse accessions were placed outside these close-knit clusters. The Bootstrap estimates obtained by Wagner parsimony analysis were significant for seven out of 49 nodes in the majority-rule consensus tree (〈95% occurrence). The results of both the analyses were, however, broadly comparable when the constitution of the individual clusters were considered. The principal components analysis indicated that the first two components accounted for only 21% of the total variations and in order to explain 〈75% of variations 18 components were required. The high level of genetic diversity prevalent among the Indian collections is probably indicative of the nativity of this crop species. Similarly, the relatively lower level of polymorphism in exotic germplasm could be ascribed to the comparatively recent introductions of limited germplasm of this crop into some of the non-traditional sesame growing countries.
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  • 79
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    Euphytica 110 (1999), S. 139-149 
    ISSN: 1573-5060
    Keywords: cultivar identification ; genetic diversity ; Prunus ; RAPD ; rootstocks
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract We have used RAPD markers to characterize Prunus rootstocks from different species, both commercial, and selected clones from the breeding program at Aula Dei Experimental Station (Zaragoza, Spain). Molecular markers were used to study the genetic variation among different species, and within species. Forty one genotypes were used in this study. They included P. amygdalo-persica, and P. persica × P. davidiana hybrids; P. cerasifera, P. domestica, and P. insititia clones, and other diverse interspecific hybrids, which were divided in three groups according to postulated taxonomic classification. Diversity patterns obtained from 80 RAPD primers were evaluated in a representative subset of genotypes. This screening helped to identify 7 RAPD primers that were selected to produce a combined classification of the whole set of rootstock clones. This analysis successfully clustered rootstocks according to the classification scheme widely used to characterize Prunus clones, mainly based on morphological descriptors. Further than that, it supported the alleged origin of some interspecific materials, and confirmed a case of possible misclassification (‘Myrobalan 29 C’). A more thorough diversity analysis was conducted within each group of materials, using larger sets of primers (12–14). After this analysis, disjointed clusters were formed for P. amygdalo-persica and P. persica × P. davidiana hybrids in one group, and for Myrobalan (P. cerasifera) and Marianna (P. cerasifera × P. munsoniana) plums in another group. P. insititia and P. domestica clones, however, formed a jumbled cluster, possibly due to genetic interchange among them during their domestication and breeding history.
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  • 80
    ISSN: 1573-5060
    Keywords: Erwinia carotovora subsp ; atroseptica ; Phytophthora infestans ; PVY ; RAPD ; Solanum brevidens ; Solanum tuberosum ; somatic hybrids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Symmetric somatic hybrids were produced by electrofusion of protoplasts of two dihaploid tuber-bearing potato (Solanum tuberosum L.) lines and Solanum brevidens Phil., a diploid non-tuber-bearing wild potato species. A total of 985 plants was obtained. Verification of nuclear hybridity of putative hybrids was based on additive RAPD patterns, general morphological characteristics and chromosome counts. 53 (90%) calli regenerated into plants which were identified as somatic hybrids. Most of the hybrids were aneuploids at the tetraploid (4×) or hexaploid (6×) level. The 20 hybrids tested expressed a high level of resistance to potato virus Y (PVY N ) characteristic of the S. brevidens parent. Resistance to late blight (Phytophthora infestans (Mont.) de Bary) varied between hybrids, but was on average better than that of the fusion parents. Resistance of hybrids to bacterial stem rot (Erwinia carotovora subsp. atroseptica (van Hall) Dye) was not superior to that of commercial potato cultivars.
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  • 81
    ISSN: 1573-5060
    Keywords: Chenopodium ; genetic relationship ; molecular markers ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The RAPD technique was used to identify genetic relationships in 19 accessions, including six species of the genus Chenopodium. A dendrogram was constructed using UPGMA from 399 DNA markers. The molecular data clustered species and accessions into five different groups. Group 1 with three cultivated varieties of C. nuttalliae, Group 2 included eight cultivars and two wild varieties of C. quinoa, Group 3 with C. berlandieri and C. album, Group 4 with two accessions of C. pallidicaule, and Group 5 with 2 accessions of C. ambrosioides. The polymorphic patterns generated by RAPD profiles showed different degrees of genetic relationship among the species studied. A low level of intraspecific variation was found within the accessions of C. quinoa, C. nuttalliae, and C. pallidicaule. The RAPD markers were found to be a useful tool for detecting genetic variation within the genus Chenopodium.
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  • 82
    ISSN: 1573-5060
    Keywords: cyst nematode ; Heterodera glycines ; diversity ; resistance ; RFLP ; RAPD ; soybean ; Glycine max
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Cultivar Peking has been extensively used as a source of resistance to Race 3 and Race 5 of soybean cyst nematode, Heterodera glycines I., and Peking genes for resistance are present in a wide range of resistant soybean cultivars. Peking is also used as a host differential in the soybean cyst nematode race classification system. Thirteen Peking lines maintained in the USDA Soybean Germplasm Collection and in several breeding programs were surveyed using RFLP and RAPD markers for genetic characterization. Based on the molecular diversity combined with reaction to soybean cyst nematode, Peking genotypes from a common original source were identified. Peking lines PI 297543 (introduction from Hungary), and PI 438496A, PI 438496B and PI 438496C (introductions from Russia) represented unrelated germplasms. Identified molecular polymorphism can be used to validate the genetic purity of Peking lines used as host differentials for soybean cyst nematode classification system as well as utilization of an individual germplasm line in genetic-breeding programs.
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  • 83
    ISSN: 1573-5117
    Keywords: aquatic plants ; RAPD ; hybridization ; genetic diversity ; Scirpus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Enzyme polymorphisms have been widely used in aquatic plants since the 1980s. Studies on DNA polymorphisms are less numerous and a case-study using both methods on Scirpus is worked out. Along the unique freshwater tidal zone of the River Schelde (Belgium), clumps of Scirpus species are mostly scattered in small and fragmented locations on the dikes and mud flats. Most of these taxa are native S. triqueter, S. tabernaemontani or intermediate morphological forms. However, several cultivated strains of S. tabernaemontani have been introduced in recent years. Such ‘exotic’ strains have been planted to stabilize the muddy riverbanks and became well established and may perform better than the native hybrid complex. In order to determine the existing genetic diversity among these species and the possibility for genetic pollution, stems of 30 clumps from a series of locations along the tidal river were investigated for seven enzymes (SDH, PGM, EST, MNR, GOT, 6PGD and ME) and for markers at DNA level using random amplified polymorphic DNA's (RAPD) of 22 decanucleotides. Data analysis of the allozymes and of the amplified DNA fragments enabled us to classify unambiguously the different Scirpus taxa. Direct evidence of hybridization between S. triqueter and S. tabernaemontani could not be obtained, but the putative hybrids are genetically intermediate or close to S. triqueterwhen considering the DNA polymorphism. The introduced clones of S. tabernaemontani consisted of at least three groups of genotypes of which one was very related to the native ones. The escaped clumps could be assigned to a third introduced but less-related strain.
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  • 84
    ISSN: 1573-5117
    Keywords: RAPD ; nile perch ; Tanganyika ; endemic ; genetic differentiation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The genetic differentiation of endemic nile perch (Lates stappersi) populations in Lake Tanganyika was studied using RAPD. DNA was extracted from alcohol stored muscle tissue by a salting method, without organic solvents. Three primers amplified 58 variable DNA fragments from 270 individuals from five localities. The genetic distances of local samples as inverse of bandsharing ranged from 0.097 to 0.312. The population sampled in Kigoma, close to the estuary of the Malagarazi river showed high values of genetic distance in pairwise comparisons with other sampled populations. Principal component analysis separated the main population and the 25 samples from Kigoma with high eigenvalues. Five individuals sampled in Kigoma were united with the main population, as confirmed by significant differences in band frequences. The local population in Kigoma had significantly different frequencies in 24 RAPD bands when compared to the pooled samples of Lates stappersi. No clearly diagnostic fragments were found. The genetic distance (1-F) between the Kigoma population and the united main stock was 0.195. Based on Slatkin's index on private alleles, the level of migration between Kigoma and all other sampling sites united, migration is restricted (Nm = 0.43) and allows genetic separation.
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  • 85
    ISSN: 1573-5117
    Keywords: DNA ; RAPD ; genetic diversity ; Bruguiera ; Sri Lanka ; mangroves
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The identification of populations of Bruguiera sexangula, Bruguiera gymnorhiza and their putative hybrids in the field is difficult using only morphological and phenological characters. Using a PCR based technique, RAPD (Random Amplified Polymorphic DNA), the genetic variation of Bruguiera populations was studied from contrasting climatic and geographic regions along the southwest coastal region of Sri Lanka. Out of 45 primers screened, 20 primers allowed us to observe polymorphism, not only between species (interspecific) but also within the species (intraspecific). Analysis of RAPD data appears to be helpful in determining the genetic relationship among populations of B. gymnorhiza and B. sexangula. RAPD markers revealed that the two species are well separated without any hybrid position between the two taxa though they occur in mixed stands. Although sampling sizes of populations of this study were small, genetic variation among B. gymnorhiza and B. sexangula populations could be observed. For B. sexangula, it was possible to differentiate each of the three populations, even when using a small number of primers.
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  • 86
    Electronic Resource
    Electronic Resource
    Springer
    World journal of microbiology and biotechnology 15 (1999), S. 381-385 
    ISSN: 1573-0972
    Keywords: Brucella abortus ; Brucella melitensis ; polymerase chain reaction ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A 1.3 kb Brucella-specific DNA fragment produced through the use of arbitrarily primed polymerase chain reaction (AP-PCR) was tested for its specificity by DNA–DNA hybridization to Brucella and non-Brucella bacteria. The digoxigenin (DIG)-labelled 1.3 kb DNA fragment hybridized with Brucella abortus and Brucella melitensis but did not hybridize with other non-Brucella bacteria tested. The sensitivity of the reaction was determined; as little as 150 fg DNA or 30 Brucella cells could be detected. The specificity and sensitivity of the 1.3 kb DNA fragment combined with the simplicity and speed of the technique suggest the potential of this fragment as a DNA probe for the quick and reliable detection of Brucella organisms.
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  • 87
    ISSN: 1573-0972
    Keywords: Bradyrhizobium japonicum ; nitrogen fixation ; RAPD ; RFLP ; strain identification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Genomic DNA of 13 Bradyrhizobium japonicum strains was prepared and analysed by restriction fragment length polymorphism (RFLP) with nif and nod probes, and by random amplified polymorphic DNA (RAPD) with 11 primers of arbitrary nucleotide sequence. Polymorphism was observed in both analyses. The RFLP and RAPD banding patterns of different strains were used to calculate genetic divergence and to construct phylogenetic trees, allowing studies on the relationships between the strains. RFLP with nif and nod probes permitted the separation of the strains into two divergent groups, whereas RAPD separated them into four main groups. RAPD allowed closely related strains to be distinguished.
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  • 88
    ISSN: 1573-8469
    Keywords: genetic variability ; plant pathogen ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Crinipellis perniciosa (Stahel) Singer is the causal agent of witches' broom disease in the Sterculiaceae, Solanaceae, and Bixaceae families. The disease is endemic to the Brazilian Amazon, and was first reported infecting Theobroma cacao (cocoa) in the State of Bahia, Brazil, in 1989. Random amplified polymorphic DNA (RAPD) analyses were performed on 46 isolates of C. perniciosa from cocoa that were collected from 15 counties in Bahia and the Brazilian Amazon. A total of 258 RAPD loci from 20 primers and three mixed primers were analyzed. Of these loci, 108 (42%) were polymorphic, with an average of 4.7 polymorphic loci per primer produced. Genetic similarities were estimated using Nei and Li's index and UPGMA clustering. Bootstrap analysis divided the phenogram into four significantly different clusters: two groups contained isolates from Ariquemes and from Ouro Preto, Rondônia, and the other two separated the isolates from Bahia into two major groups of C. perniciosa, classified as Group 1 (G1) and Group 2 (G2). The two groups of isolates from Bahia differed for their genetic similarity with the isolates from the Brazilian Amazon. The geographic distribution of the groups in Bahia suggests two independent focal points of introduction. Ongoing programs to screen for resistant cocoa genotypes should consider both groups of isolates.
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