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  • Articles  (108)
  • RAPD  (67)
  • RFLP  (49)
  • Springer  (108)
  • 1995-1999  (108)
  • 1998  (108)
  • 1
    ISSN: 1432-1890
    Keywords: Key words DNA polymorphism ; Ectomycorrhizal fungi ; Genetic diversity ; Pisolithus tinctorius ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Twenty Pisolithus tinctorius isolates from different geographic locations and different hosts were characterized by the random amplified polymorphic DNA technique. Thirteen arbitrary primers generated 87 DNA fragments, all of them polymorphic. These data were used to calculate genetic distances among the isolates. The pairwise genetic distances ranged from 1 to 100%, with an average of 58.7%. Cluster analysis based on the amplified fragments grouped the isolates according to their host and geographical origins. Group I contained isolates collected in Brazil and group II those collected in the Northern Hemisphere. In addition to the diversity seen at the molecular level, the isolates also showed host specificity. Greenhouse experiments demonstrated that isolates from the Northern Hemisphere colonized mainly Pinus whereas isolates from Brazil colonized only Eucalyptus. The molecular data suggest that the Pisolithus tinctorius isolates analyzed belong to two distinct groups. The data also suggest new guidelines for future investigations on the taxonomy and systematic of this important fungus species. Furthermore, these results support future experiments aimed at the selection and development of improved isolates of P. tinctorius.
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  • 2
    Electronic Resource
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    Springer
    Plant molecular biology reporter 16 (1998), S. 139-139 
    ISSN: 1572-9818
    Keywords: competition ; DNA mixture ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Three amplification protocols were analyzed for error rate and generation of polymorphisms during RAPD analysis. Using a set of 240 primers, the protocols detected similar frequencies of polymorphisms in two inbred sugar beet lines. The error rate was investigated by including a 1:1 mixture of DNA from the two lines in all analyses. Similar error rates, approximately 18%, were detected by the three protocols. Thus, altered amplification conditions did not substantially affect the error rate during RAPD analysis. For each of the three possible pairs of protocols, a positive correlation was obtained for primer and number of polymorphisms. Thus, a set of highly polymorphic RAPD primers can be used effectively, without prior screening, to detect polymorphisms for each protocol.
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  • 3
    ISSN: 1572-9818
    Keywords: amplified fragment length polymorphism ; cocoa ; RAPD ; woody plant
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Accurate identification of parental plants and their hybrids is essential for an effective breeding programme. Traditional classification of cocoa varieties relies on the characterisation of agricultural traits at plant maturity. A rapid and reliable method is described, based on genotypic analysis. An efficient DNA isolation procedure was developed, yielding unsheared DNA of high purity. Two genetic fingerprinting techniques, RAPD and AFLP™, were evaluated for their suitability in distinguishing cocoa varieties. RAPD analysis was unsatisfactory due to the low frequency of polymorphisms and poor reproducibility. AFLP™ was reliable in distinguishing phenotypically identical, known varieties of cocoa. Importantly, AFLP™ also revealed intra- and inter-varietal variation. Abbreviations: AFLP™, amplified fragment length polymorphism; APS, ammonium persulphate; CTAB, hexadecyltrimethylammonium bromide; DEB, DNA extraction buffer; f.wt., fresh weight; NEB, nuclei extraction buffer; PMSF, phenylmethanesulphonyl fluoride; RAPD, random amplified polymorphic DNA; T4 PNK, Bacteriophage T4 polynucleotide kinase; Taq, Thermus aquaticus; TBE, tris-borate-EDTA; TEMED, NNN′N′ tetramethylethylenediamine.
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  • 4
    ISSN: 1572-9818
    Keywords: cpDNA ; DNA extraction ; fingerprinting ; forest trees ; M13 fingerprinting ; method ; PCR ; rDNA ; RFLP ; rhododendron ; plant
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Four DNA extraction protocols were compared for ability to produce DNA from the leaves or needles of several species: oak, elm, pine, fir, poplar and maize (fresh materials) and rhododendron (silica dried or frozen material). With the exception of maize and poplar, the species are known to be difficult for DNA extraction. Two protocols represented classical procedures for lysis and purification, and the other two were a combination of classical lysis followed by anion exchange chromatography. The DNA obtained from all procedures was quantified and tested by PCR and Southern hybridisation.Test results indicated superiority of one of the four protocols; a combination of CTAB lysis followed by anion exchange chromatography which enabled DNA extraction from all seven species. A second protocol also produced DNA from leaves or needles of all species investigated and was well suited for PCR applications but not Southern hybridisations. The remaining protocols produced DNA from some but not all species tested. Abbreviations: CTAB, hexadecyltrimethylammonium bromide; EtOH, Ethanol; TBE, tris-borate-EDTA.
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  • 5
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    Plant molecular biology reporter 16 (1998), S. 91-91 
    ISSN: 1572-9818
    Keywords: Amaranthus ; DNA fingerprinting ; PCR ; polysaccharides ; RAPD ; total DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A simple, efficient and reliable method is described for isolation of total DNA from young leaves of Amaranthus species. This procedure yields a high amount (600–800 µg DNA/g fresh leaf tissue) of good quality DNA free from contaminating proteins, polysaccharides, and coloured pigments. The DNA is suitable for digestion with several restriction endonucleases, preparation of Southern blots, and PCR amplification. The DNA has been successfully used for generating DNA fingerprint profiles and RAPD banding patterns in two species of Amaranthus. The procedure is suitable for processing of a large number of samples simultaneously.
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  • 6
    ISSN: 1573-0832
    Keywords: armadillo ; Paracoccidioides brasiliensis ; PCR ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Sixty-three Paracoccidioides brasiliensis isolates obtained from three nine-banded armadillos ( Dasypus novemcinctus), one Amazonian armadillo's and 19 clinical isolates were compared by random amplified polymorphic DNA analysis with the primer OPG-19. The isolates were divided into three major clusters, I, II and III. Coincidences between human and armadillo isolates were observed in clusters I and II. Cluster III consisted only of armadillos' isolates. The results suggested that (I) humans may acquire P. brasiliensis infection by contact with armadillo's environment, (II) there may be P. brasiliensis genotypes peculiar to the animal, and (III) individual armadillos may be infected with P. brasiliensis cells with different genotypes.
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  • 7
    ISSN: 1573-0832
    Keywords: Candida ; identification ; PCR ; phylogeny ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Fast and reliable identification of different species of the genus Candida is important to define adequate therapeutic decisions, because the different species have highly variable susceptibilities to antifungal drugs; azoles and amphothericin B. Accurate statistical records on case history and epidemiological studies also depend on effective identification. To address this problem we established a RAPD method that enabled direct identification of five very common species of Candida. Initially, reference band patterns were established for C. albicans, C. tropicalis, C. parapsilosis, C. glabrata and C. krusei. One of the primers, M2, showed remarkably conserved intra-specific patterns of approximately 10 bands each, ranging in size from 2.0 to 0.1 kb. These patterns were significantly different and species-specific. Few bands were conserved between different species of Candida, which was assumed to be consistent with their phylogenetic relatedness. In addition, band patterns were constant and reproducible and DNA isolated from single colonies yielded sufficient DNA for identification. The reference band patterns were then used, in blind experiments, to identify species of Candida in 50 randomly chosen samples, including clinical isolates and ATCC strains. RAPD results were 100% consistent with results obtained by conventional diagnostic methods and were achieved in one day instead of several days taken by conventional methods. Because ideal identification methods should be consistent with phylogeny and taxonomy we tested whether RAPD could be used to calculate genetic distances. Comparison of RAPD phylogenetic trees with 18S rRNA trees showed significant differences in tree topologies which indicated that RAPD data could not accurately measure the relative distances between different species. Also, computer simulations of RAPD random patterns were used to test whether the observed degree of RAPD band pattern similarities could occur at random. These simulations suggested that the level of inter-specific band pattern similarities observed in our data could be obtained at random, while intra-specific pattern similarities could not. RAPD would be helpful to discriminate between isolates but not to quantitate the differences. We suggest that the inaccurate estimate of genetic distances from RAPD is a general limitation of the technique and not a specific problem of our identification method. Because of the repetitive character of the target sequences, genetic distances calculated from RAPD could be affected by paralogy, namely, recombination and duplication events not parallel with speciation events.
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  • 8
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    Plant molecular biology reporter 16 (1998), S. 90-90 
    ISSN: 1572-9818
    Keywords: DNA extration ; DNA fingerprint ; half seed ; PCR ; RFLP ; target gene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A single-seed DNA extraction method was developed for rapid identification of plant genotype. The method was applied to 12 plant species, including the oil seeds sesame and soybean. The results were comparable to those obtained for oil-less seeds such as rice. This method will be useful for genotypic selection which requires rapid screening of large populations. It can also be used to identify varietal purity of seed stocks by PCR and RFLP analysis. The method includes two major steps, (i) treatment by proteinase K in an SDS extraction buffer, and (ii) grinding of a single half seed in the buffer after incubation. About 1.5–2 µg of DNA per half seed (the endosperm part) of rice was obtained and more than 200 half seed samples could be handled by one person in a day. The DNA could be used for fingerprinting and detection of target genes in a transgenic plant by PCR. The amplified PCR products from the half seed DNA exhibited the same banding patterns as those from leaf DNA. Yield and quality of DNA extracted from half seeds of rice was also sufficient for RFLP analysis. The remnant half seeds containing the embryo can be maintained for later germination of selected genotypes.
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  • 9
    ISSN: 1432-1211
    Keywords: Key words IL-5Rα ; Promoter ; Polymorphism ; RFLP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
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  • 10
    ISSN: 1432-203X
    Keywords: Key words Somatic hybridization ; RAPD ; Citrus huanglongbin ; Sexual and graft incompatibility ; Aurantioideae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Protoplasts isolated from `Page' tangelo (Minneola tangelo × clementine) cell suspension cultures were electrically fused with mesophyll protoplasts of orange jessamine [Murraya paniculata (L.) Jack]. Shoots were regenerated after 6 – 10 months of culture, but they were extremely recalcitrant to producing roots in root-induction medium. Complete plantlets were formed via micrografting. Chromosome counting of shoot tips revealed they were tetraploids (2n = 4x = 36). Glutamateoxaloacetate transaminase isozyme and randomly amplified polymorphic DNA analysis confirmed their hybridity. Orange jessamine is immune to citrus huanglongbin, a severe disease of citrus, but sexual incompatibility and limited graft compatibility exist between Citrus and orange jessamine. The cell fusion technique may make it possible to transfer the huanglongbin resistance trait from orange jessamine to Citrus.
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  • 11
    ISSN: 1432-203X
    Keywords: Key words Genetic stability ; Micropropagation ; Pinewood-nematode ; Pinus thunbergii ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Random amplified polymorphic DNA (RAPD) markers were used to determine the genetic stability of long-term (more than 10 years) micropropagated shoots of Japanese black pine (Pinus thunbergii Parl.). Thirty-six shoots consisting of three morphotypes (short, medium, and long needles) were randomly chosen from about 4,000 micropropagated shoots regenerated from the explants of a single nematode-resistant mother plant. Out of 126 primers screened, 30 gave 134 clear reproducible bands. A total of 4,824 bands obtained from these studies exhibited no aberration in RAPD banding patterns among the tested shoots. Our results show that regenerants from our plant micropropagation system are genetically stable.
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  • 12
    ISSN: 1432-2145
    Keywords: Key words Abies ; Egg cell ; Plastid inheritance ; RFLP ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The ultrastructure of egg cells in Abies alba was examined to elucidate the lack of maternal inheritance of plastids. Before fertilization, maternal plastids are absent in the perinuclar zone containing mainly mitochondria and smooth endoplasmic reticulum. During egg cell development the maternal plastids are transformed into large inclusions which are situated mostly towards the periphery of the egg cell, and finally disintegrate. As a consequence, they do not participate in zygote formation. RFLP analysis of cpDNA of parental trees and their F1 interspecific hybrids (A. alba×A. numidica, A. alba×A. nordmanniana, A. nordmanniana×A. Alba) using HindIII and BamHI showed a paternal mode of cpDNA inheritance. Paternal inheritance has also been found with PCR/RFLP analysis of cpDNA from parental trees and their hybrids (A. alba×A. pinsapo, A. pinsapo×A. alba, A. pinsapo×A. numidica) using ApaI and HaeIII digests, as well as in the crosses of A. cephalonica×A. nordmanniana, A. nordmanniana×A. cephalonica, A. cephalonica×A. numidica using TagI digests.
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  • 13
    ISSN: 1432-203X
    Keywords: Key wordsPopulus alba L. ; Protoplast ; Plant regeneration ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We developed an efficient plant regeneration system from protoplasts for poplar (Populus alba L.). Protoplasts were isolated from 4-day-old suspension cultures derived from seed-induced calli with a yield of 6.96× 106 cells/g fresh weight cells and then cultured at a concentration of 2.5×105 cells/ml in NH4NO3-free Murashige and Skoog (MS) medium supplemented with 5 µM 2,4-dichlorophenoxyacetic acid (2,4-D), 0.05 µM thidiazuron (TDZ) and 0.5 M glucose as a osmoticum. The plating efficiency of the cultured protoplasts was calculated at 26.5% at day 7 and 31.7% at day 14. Cell colonies were observed after culturing for 4 weeks. Regenerated colonies were propagated through subculture in liquid MS medium supplemented with 5 µM 2,4-D. Buds were induced from regenerated calli on MS medium containing 10 µM kinetin or 1 µM TDZ. Regenerated shoots were rooted on half-strength MS medium, and the plantlets were transplanted in soil. Randomly amplified polymorphic DNA analysis did not detect any DNA polymorphism among the regenerated plants.
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  • 14
    ISSN: 1432-203X
    Keywords: Key words DNA content ; Morphology ; Protoplast fusion ; RAPD ; Somatic hybrid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Solanum acaule Bitt. is a disomic tetraploid (4x) wild potato species which is resistant to several potato diseases. Introgression of disease resistance and abiotic stress tolerance to the tetrasomic tetraploid (4x) cultivated potato (S. tuberosum L.) gene pool via crossing has been limited due to the difference in the endosperm balance number. In the present study, protoplast fusion was applied to produce hexaploid (6x) somatic hybrids between the parental lines, tetraploid (4x) S. acaule and two anther-derived dihaploid (2x) lines of S. tuberosum cv. White Lady. One callus (0.4%) of a total of 229 calli obtained regenerated into shoots in the fusion combination S. acaule (+) White Lady 15.dh.8.2.2. All the regenerated shoots were confirmed to be interspecific somatic hybrids using species-specific RAPD markers. In another fusion combination, S. acaule (+) White Lady 7.dh.23.1.1, fifteen calli (5%) regenerated into a total of sixteen shoots from 289 calli. All the analysed somatic hybrids between S. acaule and S. tuberosum were hexaploid. The mean DNA content (2C value) of the combination S. acaule (+) White Lady 15.dh.8.2.2 somatic hybrids (4.55 pg), was approximately the sum (4.69 pg) of the DNA contents of the parental lines, S. acaule (2.95 pg) and S. tuberosum (1.74 pg). In the greenhouse, the two somatic hybrids analysed were normal in their morphological characteristics and more vigorous than their parental lines. Most of the morphological characteristics were closer to the tetraploid S. acaule than to the dihaploid S. tuberosum. The interspecific somatic hybrids are currently being tested for frost tolerance and glycoalkaloid composition.
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  • 15
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    Theoretical and applied genetics 97 (1998), S. 1321-1330 
    ISSN: 1432-2242
    Keywords: Key words Maize ; Ustilago maydis ; QTL mapping ; RFLP ; Resistance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  We mapped and characterized quantitative trait loci (QTLs) for resistance to Ustilago maydis and investigated their consistency across different flint-maize populations. Four independent populations, comprising 280 F3 lines (A×BI), 120 F5 lines (A×BII), 131 F4 lines (A×C) and 133 F4 lines (C×D), were produced from four European elite flint inbreds (A, B, C, D) and genotyped at 89, 151, 104, and 122 RFLP marker loci, respectively. All Fn lines were evaluated in field trials with two replications in five German environments. Genotypic variances were highly significant for the percentage of U. maydis infected plants (UST) in all populations, and heritabilities exceeded 0.69. Between five and ten QTLs were detected in individual populations by composite interval mapping, explaining between 39% and 58% of the phenotypic variance. These 19 different QTLs were distributed over all ten chromosomes without any clustering on certain chromosomes. In most cases, gene action was dominant or overdominant. Fourteen pairs of the detected QTLs for UST displayed significant digenic epistatic interactions, but only two of them did so after arcsin √UST/100 transformation. Significant QTL× environment interactions occurred frequently. Between two to four QTLs were common between pairs of populations. Population C×D was also grown in Chartres, a location with a high U. maydis incidence. Two out of six QTLs identified for Chartres were in common with QTLs detected across five German environments for C×D. Consequently, marker-assisted or phenotypic selection based on results from natural infection seem to be suitable breeding strategies for improving the resistance of maize to U. maydis.
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  • 16
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    Theoretical and applied genetics 96 (1998), S. 31-36 
    ISSN: 1432-2242
    Keywords: Key words Foxtail millet ; Genetic map ; Primary trisomics ; RFLP ; Setaria italica ; Setaria viridis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  An RFLP-based map consisting of 160 loci was constructed in an intervarietal cross of foxtail millet [Setaria italica (L.) P. Beauv.], Longgu 25×Pagoda Flower Green. The map comprises nine linkage groups, which were aligned with the nine foxtail millet chromosomes using trisomic lines, and spans 964 cM. The intraspecific map was compared to an interspecific map, constructed in a S. italica×S. viridis cross. Both the order of the markers and the genetic distances between the loci were highly conserved. Deviations from the expected 1 : 2 : 1 Mendelian segregation ratios were observed in both the intra- and inter-specific populations. The segregation data indicate that chromosome VIII in the Longgu 25×Pagoda Flower Green cross carries a gene that strongly affects gamete fertility.
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  • 17
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    Theoretical and applied genetics 96 (1998), S. 101-111 
    ISSN: 1432-2242
    Keywords: Key words Hordeum ; Barley ; RAPD ; Variability ; Phylogeny ; DNA analyses
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The genetic variation of 102 natural populations of wild barley growing in Spain was assessed using RAPDs (random amplified polymorphic DNA). The plant material included the annual species H. marinum subsp. marinum (22 populations) and subsp. gussoneanum (14), H. murinum subsp. murinum (7) and subsp. leporinum (35), and the perennial species H. bulbosum (17) and H. secalinum (7). Ten of the tested 64 arbitrary 10-mer primers amplified polymorphic DNA in all taxonomic units. Analyses was performed within and between populations, species and subspecies. The primers gave a total of 250 RAPD products. The level of polymorphism varied between taxonomic units depending on the primers employed and the plant reproductive system. In general, the most variable were the allogamous species H. secalinum and H. bulbosum and the autogamous H. marinum subsp. marinum. Among the amplified bands, 69 (27%) were shared by at least two different taxonomic units. The remaining bands were specific. The results demonstrate differences in the degree of similarity between taxonomic units. Jaccard’s similarity coefficients for interval measure within and between populations were used to produce a cluster diagram using the unweighted pair-group method (UPGMA). The different populations of the species and subspecies of Hordeum fell into three groups. The first group contained the populations belonging to both subspecies of H. marinum, plus those of H. secalinum. The populations of H. marinum subsp. gussoneanum were very closely associated. Those of H. marinum subsp. marinum were grouped in a broad cluster. The second group, occupying the innermost position of the tree, was very closely associated with the populations of both subspecies of H. murinum. The third branch segregated H. bulbosum. A series of RAPD markers were investigated by cleaving the amplified products of the same size with restriction endonucleases that recognize targets of 4- or 6-bp. The production of equivalent fragments following cleavage by the same enzyme would seem to demonstrate their homology in samples from different individuals, populations or taxonomic units.
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  • 18
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    Theoretical and applied genetics 96 (1998), S. 468-474 
    ISSN: 1432-2242
    Keywords: Key words Microsynteny ; Genome ; Gene cluster ; RFLP ; PFGE
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Genetic and physical maps, consisting of a large number of DNA markers for Arabidopsis thaliana chromosomes, represent excellent tools to determine the organization of related genomes such as those of Brassica. In this paper we report the chromosomal localization and physical analysis by pulsed-field gel electrophoresis (PFGE) of a well-defined gene complex of A. thaliana in the Brassica nigra genome (B genome n=8). This complex is approximately 30 kb in length in A. thaliana and contains a cluster of six genes including ABI1 (ABA-responsive), RPS2 (resistance against Pseudomonas syringae, a bacterial disease), CK1 (casein kinase I), NAP (nucleosome-assembly protein), X9 and X14 (both of unknown function). The Arabidopsis chromosomal complex was found to be duplicated and conserved in gene number at different levels in the Brassica genome. Linkage group B1 had the most-conserved arrangement carrying all six genes tightly linked. Group B4 had an almost complete complex except for the absence of RPS2. Other partial complexes of fewer members were found on three other chromosomes. Our studies demonstrate that by this approach it is possible to identify ancestrally related chromosome segments in a complex and duplicated genome, such as the genome of B. nigra, permitting one to draw conclusions as to its origin and evolution.
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  • 19
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    Theoretical and applied genetics 96 (1998), S. 683-687 
    ISSN: 1432-2242
    Keywords: Key words Essential oils ; Mint ; Mentha piperita ; M. spicata ; Somatic hybridization ; Plant regeneration ; RAPD ; Southern hybridization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Twenty eight somatic hybrid plants were identified following protoplast fusions between peppermint (Mentha piperita L. cv Black Mitcham), producing high-quality oil, and spearmint (Mentha spicata L. cv Native Spearmint), likewise producing high-quality oil and also possessing resistance to verticillium wilt. Prior to fusion, peppermint protoplasts were subjected to iodoacetic acid to inhibit cell division. Protoplasts of peppermint and spearmint were fused using polyethylene glycol plus DMSO. Fusion products were cultured according to an efficient protoplast-to-plant-cycle protocol developed for peppermint. Using this protocol, iodoacetic acid-treated peppermint protoplasts were not able to divide, whereas untreated spearmint protoplasts had the ability to produce callus but not shoots. Therefore, selection of somatic hybrid calli was based on the presumed capability of hybrid cells to form calli and shoots. Shoots in vitro were initially identified as hybrids using RAPD profiles. Subsequently, observations on morphology, chromosome counts, and Southern-hybridization patterns confirmed their hybrid status. The results of verticillium tests revealed that 18 somatic hybrids were more susceptible than Native Spearmint, while hybrid II-14 had a level of susceptibility intermediate between that of the fusion parents. Oil-analysis of hybrid plants indicated that they all have a GC-profile typical of spearmint oil.
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  • 20
    ISSN: 1432-2242
    Keywords: Key words Medicago sativa ; RAPD ; Cultivars ; Genetic distance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Alfalfa (Medicago sativa L.) is a forage legume of world-wide importance whose both allogamous and autotetraploid nature maximizes the genetic diversity within natural and cultivated populations. This genetic diversity makes difficult the discrimination between two related populations. We analyzed this genetic diversity by screening DNA from individual plants of eight cultivated and natural populations of M. sativa and M.  falcata using the RAPD method. A high level of genetic variation was found within and between populations. Using five primers, 64 intense bands were scored as present or absent across all populations. Most of the loci were revealed to be highly polymorphic whereas very few population-specific polymorphisms were identified. From these observations, we adopted a method based on the Roger’s genetic distance between populations using the observed frequency of bands to discriminate populations pairwise. Except for one case, the between-population distances were all significantly different from zero. We have also determined the minimal number of bands and individuals required to test for the significance of between-population distances.
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  • 21
    ISSN: 1432-2242
    Keywords: Key words Mangroves ; Genome relationship ; Phylogeny ; RAPD ; RFLP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  DNA from pooled leaf samples of 11 true major mangrove, three true minor mangrove, two mangrove associate, two mangrove parasite, three terrestrial and one cultivated species were isolated for the present study. In total, 198 random amplified polymorphic DNAs (RAPDs) and 180 restriction fragment length polymorphism (RFLP) loci were scored by using ten primers and 14 enzyme-probe combinations respectively. The polymorphism observed for these markers revealed a high degree of genetic diversity in mangroves at both inter-specific or inter-generic levels. A dendrogram, constructed after pooling both RAPD and RFLP data, using a similarity index was analysed for genome relationships among these species. The dendrogram showed clustering of all the major mangroves, except for Nypa fruticans (Arecaceae), into one group. All species under the tribe Rhizophorae formed a sub-cluster, to which Xylocarpus granatum was found to be the most closesly related species. The clustering pattern implied that Excoecaria agallocha and Acanthus ilicifolius should be considered as true minor mangroves. The present study also provided molecular data favouring the separation of Avicennia spp. from the Verbenaceae to create a monotypic family the Avicenniaceae. The separation of Viscum orientale into the Viscaceae was also favoured.
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  • 22
    ISSN: 1432-2242
    Keywords: Key words Durum wheat ; Mapping ; Linkage map ; RFLP ; Morphological markers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  A genetic linkage map of tetraploid wheat [Triticum turgidum (L.) Thell.] was constructed using segregation data from a population of 65 recombinant inbred lines (RILs) derived from a cross between the durum wheat cultivar Messapia and accession MG4343 of T. turgidum (L.) Thell. ssp dicoccoides (Korn.) Thell. A total of 259 loci were analysed, including 244 restriction fragment length polymorphisms (RFLPs), one PCR (polymerase chain reaction) marker (a sequence coding for a LMW (low-molecular-weight) glutenin subunit gene located at the Glu-B3 locus), seven biochemical (six seed-storage protein loci and one isozyme locus) and seven morphological markers. A total of 213 loci were mapped at a LOD≥3 on all 14 chromosomes of the A and B genomes. The total length of the map is 1352 cM and the average distance between adjacent markers is 6.3 cM. Forty six loci could not be mapped at a LOD≥3. A fraction (18.6%) of the markers deviated significantly from the expected Mendelian ratios; clusters of loci showing distorted segregation were found on chromosomes 1B, 3AL, 4AL, 6AL and 7AL. The durum wheat map was compared with the published maps of bread wheat using several common RFLP markers and general features are discussed. The markers detected the known structural rearrangements involving chromosomes 4A, 5A and 7B as well as the translocation between 2B-6B, but not the deletion on 2BS. This map provides a useful tool for analysing and breeding economically important quantitative traits and for marker-assisted selection, as well as for studies of genome organisation in small grain cereal species.
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  • 23
    ISSN: 1432-2242
    Keywords: Key words RFLP ; RAPD ; Genetic map ; Null loci ; Gene family
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  We have constructed a sex-averaged genetic linkage map in coastal Douglas-fir (Pseudotsuga menziesii [Mirb.] Franco var ‘menziesii’) using a three-generation outcrossed pedigree and molecular markers. Our research objectives are to learn about genome organization and to identify markers associated with adaptive traits. The map reported here is comprised of 141 markers organized into 17 linkage groups and covers 1,062 centiMorgans (cM). Of the markers positioned on the map, 94 were derived from a Douglas-fir complimentary-DNA (cDNA) library that was constructed from new-growth needle tissue. Other markers include 11 Douglas-fir genomic-DNAs, 20 loblolly pine (Pinus taeda L.) cDNAs, 15 random amplified polymorphic DNAs (RAPDs) and a PCR-amplified phytochrome probe. A high degree of variation was detected in each of the two parents of our mapping population, and many of the restriction fragment length polymorphism (RFLP) and RAPD phenotypes were complex. Marker data were analyzed for linkage using mapping software JOINMAP version 2.0.
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  • 24
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    Theoretical and applied genetics 97 (1998), S. 1000-1012 
    ISSN: 1432-2242
    Keywords: Key words CCN ; RFLP ; PCR ; Heterodera avenae ; Genetic mapping
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  A genetic map of the long arm of chromosome 6R of rye was constructed using eight homoeologous group-6 RFLP clones and five PCR markers derived from the rye-specific dispersed repetitive DNA family, R173. The map was developed using a novel test-cross F1 (TC-F1) population segregating for resistance to the cereal cyst nematode. Comparisons were made between the map generated with other rye and wheat group-6 chromosome maps by the inclusion of RFLP clones previously mapped in those species. Co-linearity was observed for common loci. This comparison confirmed a dramatic reduction in recombination for chromosome 6R in the TC-F1 population. The CreR locus was included in the linkage map via progeny testing of informative TC-F1 individuals. CreR mapped 3.7 cM distal from the RFLP locus, XksuF37. Comparative mapping should allow the identification of additional RFLP markers more closely linked to the CreR locus.
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  • 25
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    Theoretical and applied genetics 96 (1998), S. 791-796 
    ISSN: 1432-2242
    Keywords: Key words Sorghum ; RAPD ; CMS ; Fertility restoration ; MtDNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Seven sorghum restorer lines that differentially restore (or maintain) the A1 and A2 cytoplasmic male-sterile (CMS) cytoplasms were studied by RFLP analyses of their mtDNAs and RAPD analyses of their mitochondrial DNA (mtDNA) and total DNA to understand nuclear mitochondrial combinations that are present in these lines. RFLP data from 11 mitochondrial gene probes were inadequate to classify these seven lines. However, the analysis of RAPD profiles of total DNA could distinguish these lines on the basis of their ability to restore completely or partially the fertility in the A1/A2 CMS cytoplasms. Interestingly, RAPD profiles of mtDNAs of these lines also followed the same pattern as that of the total DNA. These results indicate that the different restorer lines possess specific nuclear-cytoplasm combinations. Further, the results also show that the RAPD technique can be used to identify markers for different cytoplasms used in CMS.
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  • 26
    ISSN: 1432-2242
    Keywords: Key words Soybean ; Glycine max ; QTL ; RFLP ; Antixenosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  One hundred and thirty nine restriction fragment length polymorphisms (RFLPs) were used to construct a soybean (Glycine max L. Merr.) genetic linkage map and to identify quantitative trait loci (QTLs) associated with resistance to corn earworm (Helicoverpa zea Boddie) in a population of 103 F2-derived lines from a cross of ‘Cobb’ (susceptible) and PI229358 (resistant). The genetic linkage map consisted of 128 markers which converged onto 30 linkage groups covering approximately 1325 cM. There were 11 unlinked markers. The F2-derived lines and the two parents were grown in the field under a plastic mesh cage near Athens, Ga., in 1995. The plants were artificially infested with corn earworm and evaluated for the amount of defoliation. Using interval-mapping analysis for linked markers and single-factor analysis of variance (ANOVA), markers were tested for an association with resistance. One major and two minor QTLs for resistance were identified in this population. The PI229358 allele contributed insect resistance at all three QTLs. The major QTL is linked to the RFLP marker A584 on linkage group (LG) ‘M’ of the USDA/Iowa State University public soybean genetic map. It accounts for 37% of the total variation for resistance in this cross. The minor QTLs are linked to the RFLP markers R249 (LG ‘H’) and Bng047 (LG ‘D1’). These markers explain 16% and 10% of variation, respectively. The heritability (h2) for resistance was estimated as 64% in this population.
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  • 27
    ISSN: 1432-2242
    Keywords: Key words Tomato and Solanum lycopersicoides intergeneric hybrid ; Chloroplast DNA ; Nuclear genome ; RFLP ; GISH
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  RFLP (restriction fragment length polymorphism) and GISH (genomic in situ hybridization) analyses were employed to identify the chloroplast and nuclear genomes of the somatic hybrids and progeny between tomato ‘Ohgata zuiko’ and Solanum lycopersicoides (‘LA 2386’). A random distribution of the chloroplast genotype was determined using a cloned 19.6-kb BamHI fragment (Ba1) of tobacco chloroplast DNA. Eight selected hybrids were analyzed for their chromosomal compositions; 4 were tetraploids (2n=48) with an equal number of chromosomes derived from each parent as accurately determined by GISH, and the other 4 were hexaploids, containing an average of two sets of tomato chromosomes and one set from the wild parent. RFLP analysis with six tomato nuclear probes of known chromosomal locations revealed no major variation among the 44 hybrid plants surveyed. However, it also showed the presence of both parent-specific alleles and the loss of some and the presence of a few non-parental alleles, indicating rearrangement and/or recombination of the nuclear DNA. The relevance of the molecular and cytological methods and the potential use of somatic hybrids for plant breeding are demonstrated.
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  • 28
    ISSN: 1432-2242
    Keywords: Key words Potato virus A ; Potato virus Y ; Resistance gene ; RFLP ; Diploid potato
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The gene Ry adg that confers resistance to potato Y potyvirus (PVY) in the cultivated potato [Solanum tuberosum subsp. andigena, line 2x(v-2)7] is located on chromosome XI in a segment that contains three other known resistance genes in other syntenic solanaceous species. One of them is the gene N that controls resistance to tobacco mosaic tobamovirus in tobacco and has previously been isolated and sequenced. Three sequence-related, resistance gene-like (RGL) DNA fragments (354–369 bp) highly homologous to the gene N were PCR-amplified from the potato line 2x(v-2)7. Two RGL fragments (79 and 81% homologous to the N gene) co-segregated with Ry adg among the 77 F1 progeny tested. These RGLs may originate from a resistance gene family on chromosome XI. The potato line 2x(v-2)7 also expressed resistance to potato A potyvirus (PVA), which was controlled by another locus on chromosome XI mapped ca. 6.8 cM distal to Ry adg .
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  • 29
    ISSN: 1432-2242
    Keywords: Key words Brassica napus ; Raphanus sativus ; Restorer gene ; Introgression ; RFLP ; RAPD ; Genetic mapping
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Bulked segregant analysis and comparative mapping were applied to identify molecular markers linked to the Rfo restorer gene used for the Ogu-INRA cytoplasmic male-sterility system in rapeseed. These markers were then used to localise the radish introgression on the B. napus genetic map constructed from the cross ‘Darmor.bzh’ x ’Yudal’. The introgression mapped on the DY15 linkage group. From the comparison of this latter group to the linkage group constructed on a F2 progeny segregating for the radish introgression, it was concluded that the introgression had occurred through homoeologous recombination, that it was not distal and that it had replaced a B. napus region of around 50 cM. A QTL involved in aliphatic seed glucosinolate content was located on the DY15 linkage group at a position corresponding to one end of the introgression. The DNA markers identified in this study are being used in map-based cloning of the Rfo gene and in marker-assisted selection.
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  • 30
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    Theoretical and applied genetics 97 (1998), S. 147-153 
    ISSN: 1432-2242
    Keywords: Key words Genetic mapping ; Isozyme ; RFLP ; Secale cereale L. ; Self-fertility
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Three mutations determining self-fertility at the S, Z and S5 self-incompatibility loci on chromosomes 1R, 2R and 5R of rye, respectively, were mapped using three different F2 populations. There was a close linkage of one isozyme and four RFLP markers, and no recombinant plants were detected. These markers are Prx7, Xiag249 and Xpsr634 for the S locus (1R), Xbcd266 for the Z locus (2R) and Xpsr100 for the S5 locus (5R). Linkage data for markers associated to the self-fertility mutations at the S, Z and S5 loci were calculated and compared with genetic maps computed by MAPMAKER multipoint analysis.
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  • 31
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    Theoretical and applied genetics 97 (1998), S. 238-245 
    ISSN: 1432-2242
    Keywords: Key words Wheat ; Flour colour ; QTL mapping ; RFLP ; AFLP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  An RFLP map constructed using 150 single seed descent (SSD) lines from a cross between two hexaploid wheat varieties (‘Schomburgk’בYarralinka’) was used to identify loci controlling flour colour. Flour colour data were obtained from field trials conducted over two seasons at different sites. The estimated heritability of this trait was calculated as 0.67. Two regions identified in the preliminary analysis on chromosomes 3A and 7A, accounted for 13% and 60% of the genetic variation respectively. A detailed analysis of the major locus on 7A was conducted through fine mapping of AFLP markers identified using bulked segregant analysis (BSA). Seven additional markers were identified by the BSA and mapped to the region of the 7A locus. The applicability of these markers to identify wheat lines with enhanced flour colour is discussed.
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  • 32
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    Theoretical and applied genetics 97 (1998), S. 797-800 
    ISSN: 1432-2242
    Keywords: Key words Genetic mapping ; RFLP ; Flowering time ; Photoperiod response ; Barley
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The gene ea 7 determining photoperiod insensitivity under short day length was mapped on the short arm of chromosome 6H near the centromere. The gene was linked to the two flanking markers Xmwg2264 and Xmwg916 by 6.7 and 13.0 cM, respectively. Compared to Ppd-H1 (chromosome 2H) and Ppd-H2 (chromosome 1H), ea 7 determines the strongest effect on flowering time with 55 and 18 days difference compared to photoperiod sensitive genotypes grown under short and long photoperiods, respectively. Allelic and homoeologous relationships to major genes and quantitative trait loci controlling flowering time in barley and wheat are discussed.
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  • 33
    ISSN: 1432-2242
    Keywords: Key words CAP ; py-1 ; RAPD ; RFLP ; Breeding
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  We report the molecular mapping of the py-1 gene for resistance to corky root rot [Pyrenochaeta lycopersici (Schneider and Gerlach)] in tomato using RAPD and RFLP marker analysis. DNA from near-isogenic lines (NILs) of tomato differing in corky root rot resistance was screened with 575 random oligonucleotide primers to detect polymorphic DNAs linked to py-1. Three primers (OPW-04, OPC-02, OPG-19) revealed polymorphisms between the NILs. Twelve resistant and eight susceptible DNA pools derived from segregating F3 families were used to confirm that the RAPD markers were linked to the py-1 gene. Two of the linked amplified fragments, corresponding to OPW-04 and OPC-02, were subsequently cloned and mapped on the tomato molecular linkage map as RFLPs. These clones were located between TG40 and CT31 on the short arm of chromosome 3. Further analysis with selected RFLP markers showed that 7% (8.8 cM) of chromosome 3 of the resistant line ‘Moboglan’ was introgressed from the L. peruvianum donor parent. Three RFLP markers (TG40, TG324, and TG479) from the introgressed part of chromosome 3 were converted to cleaved amplified polymorphism (CAP) markers for use in a polymerase chain reaction (PCR) assay. These PCR markers will allow rapid large-scale screening of tomato populations for corky root rot resistance.
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  • 34
    ISSN: 1432-2242
    Keywords: Key words Apical dominance ; Bulk segregant analysis ; Map ; Pea ; RAPD ; SCAR
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Random amplified polymorphic DNA (RAPD) markers linked to two morphological markers ( fa and det), three ramosus genes (rms2, rms3 and rms4) and two genes conferring flowering response to photoperiod in pea (sn, dne) were selected by bulk segregant analysis on F2 populations. Two RAPD fragments were cloned and sequenced to generate the two SCAR markers V20 and S2 which are linked to rms3 and dne, respectively. All these genes, except rms2, were previously located on the pea classical linkage map. Rms2 mapped to linkage group IB which contains the afila gene. Precise genetic maps of the regions containing the genes were obtained and compared to the RAPD map generated from the recombinant inbred-lines population of the cross Térèse×K586. This cross was chosen because several mutants were obtained from cultivars Térèse and Torsdag (K586 was derived from Torsdag). This collection of isogenic lines was used for the construction of F2 mapping populations in which polymorphic RAPD markers were already known and mapped. Moreover, the well-known problem in pea of variability in the linkage associations between crosses was avoided. This work contributes to the precise integration between the classical map and the molecular maps existing in pea.
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  • 35
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    Theoretical and applied genetics 97 (1998), S. 1210-1216 
    ISSN: 1432-2242
    Keywords: Key words Soybean ; Glycine max ; RFLP ; QTL ; Plant height ; Canopy width
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  There are both economic and environmental reasons for reducing the use of herbicides for weed control in soybean [Glycine max (L.) Merr.] fields. Optimizing crop competitiveness can reduce reliance on chemical weed control. Fast and vigorous early growth and rapid canopy development can be effective in suppressing weed infestation of crop plants. The purposes of this study were to identify and molecularly map the quantitative trait loci (QTLs) conditioning soybean plant height and canopy width during the early vegetative stages of soybean growth. A restriction fragment length polymorphism (RFLP) linkage map was created using 142 markers and 116 F2-derived lines from a cross of ‘S100’בTokyo’. The parents and the 116 F2-derived lines were evaluated in the greenhouse and in the field at Athens, Ga., in 1996 and 1997. Combined over environments, Tokyo averaged 41 and 17% taller plants than S100 at the V7 and V10 stages of development. Transgressive segregation was observed among the progeny at both stages. Based on single-factor analysis of variance (ANOVA), three and four independent RFLP loci were associated with plant height at the V7 and V10 stages, respectively. All three loci detected [on linkage groups (LGs) C2 and F, and unlinked] at the V7 stage were also detected at the V10 stage along with one additional independent locus on LG E. The Tokyo allele contributed to increased plant height at all loci except at the unlinked locus. Three QTLs (on LGs C2, E, and F) were consistent across environments, three (on LGs C2 and F, and unlinked) were consistent across stages of plant development, and two (on LGs C2 and F) were consistent both across environments and stages of plant development. Within each stage of development, there was no interaction among the independent loci, and the respective loci together explained most of the variation in the traits. Three independent RFLP loci were associated with canopy width at the V10 stage, of which one was unique to the trait, while the remaining loci (on LGs C2 and F) were in common with the independent loci for plant height. Canopy width had a strong correlation (r=0.87) with plant height at the V10 stage. However, mature plant height, lodging, or seed weight had no phenotypic or QTL association with early plant height or canopy width.
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  • 36
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    Theoretical and applied genetics 97 (1998), S. 1279-1288 
    ISSN: 1432-2242
    Keywords: Key words Consensus map ; Isozymes ; RFLP ; Rye ; Secale cereale L.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Consensus linkage maps were constructed for all seven rye chromosomes using 12 basic RFLP maps. The maps presented contain a total of 413 markers. The number of markers per chromosome varies from 41 (chromosome 3R) to 83 (chromosome 1R). In addition to 374 RFLP and 24 isozyme markers 15 gene loci were incorporated, determining the traits reduced plant height, self fertility, male sterility restoration, vernalization response, resistance against powdery mildew, chlorophyll deficiency, hairy leaf sheath, hairy peduncle, waxy endosperm, waxless plant and absence of ligules. The maps presented allow the selection of markers for the fine mapping of certain regions of the rye genome. In terms of the known chromosomal rearrangements within the Triticeae its utilization can also be extended for mapping in wheat and barley.
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  • 37
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    Theoretical and applied genetics 96 (1998), S. 37-45 
    ISSN: 1432-2242
    Keywords: Key words Microsatellite ; RAPD ; PCR ; Linkage map ; Wheats
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The potential of PCR-based markers for construction of a genetic linkage map in Einkorn wheat was investigated. From a comparison of polymorphisms between two Einkorn wheats, Triticum monococcum (Mn) and T. boeoticum (Bt), we obtained 49 polymorphic bands produced by 33 primers for inter-simple sequence repeat (ISSR) and 36 polymorphic bands shown by 25 combinations of random amplified polymorphic DNA (RAPD) primers for mapping in 66 individuals in the F2 population. Although 44 ISSR fragments and 29 RAPD fragments statistically showed a 3 : 1 segregation ratio in the F2 population, only 9 markers each of the ISSR and RAPD bands were able to be mapped on the RFLP linkage map of Einkorn wheat. ISSR markers were distributed throughout the chromosomes. The mapped positions of the ISSR markers seemed to be similar to those obtained by the RFLP markers. On the other hand, 4 of the 9 RAPD markers could map the RFLP marker-poor region on the short arm of 3Am, suggesting a potential to map novel regions containing repetitive sequences. Comparisons of the genetic linkage map of Einkorn wheat to the linkage map and cytological map of common wheat revealed that the marker orders between the two maps of Einkorn wheat and common wheat coincided except for 4A, which harbors chromosome rearrangements specific for polyploid wheats, indicating a conservatism between the two genomes. Recombinations in Einkorn wheat chromosomes took place more frequently around the centromere and less at the distal part of chromosomes in comparison to those in common wheat. Nevertheless, recombinations even in Einkorn wheat chromosomes were strongly suppressed around the centromere. In fact, the markers located within 1 cM of the centromere were located almost in the central part of the chromosome arm.
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  • 38
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    Theoretical and applied genetics 97 (1998), S. 950-959 
    ISSN: 1432-2242
    Keywords: Key words Vitis vinifera. L ; Seedlessness ; RAPD ; SCAR ; BSA ; Marker-assisted selection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The variety Vitis vinifera cv Sultanine presents a type of seedlessness in which fertilization occurs but seeds subsequently fail to develop. It has been suggested that this trait might be controlled by three complementary recessive genes regulated by a dominant gene named I. Bulk segregant analysis was used to search for random amplified polymorphic DNA (RAPD) markers linked to the I gene in progeny obtained by crossing two partially seedless genotypes. One hundred and forty decamer primers were screened using bulks obtained by pooling the DNA of extreme individuals from the phenotypic distribution. We identified two RAPD markers which appeared tightly linked to I (at 0.7 and 3.5 cM respectively). The closest marker was used to develop a codominant SCAR (sequence characterized amplified region), named SCC8. This latter marker appeared of great value either to exclude from the progeny potentially seeded individuals or to select for seedless individuals. Indeed, all the seeded individuals of the progeny were found to be homozygous scc8 -/scc8 -, and all the individuals homozygous SCC8 +/SCC8 + were seedless. Moreover, this marker was successfully applied to other natural seedless varieties where codominance persisted. SCC8 was also used to dissect more precisely the genetics of seedlessness. ANOVA analysis indicated that this SCAR marker accounted for at least 64.9% of the phenotypic variation of the seed’s fresh weight and for at least 78.7% of the phenotypic variation of the seed’s dry matter. These results confirmed the presence of a major gene, and also the existence of other complementary recessive genes, controlling the expression of seedlessness.
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  • 39
    ISSN: 1432-2242
    Keywords: Key words Chromosome addition line ; Synteny group ; Brassica campestris ; Brassica oxyrrhina ; Monosomic ; Alloplasmic ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Monosomic chromosome addition lines of Brassica oxyrrhina in the background of alloplasmic B. campestris carrying B. oxyrrhina cytoplasm were generated and characterised through morphology, cytology and molecular (RAPD) analysis. Four successive backcrosses of the synthetic alloploid B. oxycamp with B. campestris yielded 24 monosomic addition plants that were grouped into seven different synteny groups based on morphological similarity and RAPD patterns. Each synteny group exhibited morphological features diagnostic for the presence of individual B. oxyrrhina chromosomes including some novel phenotypes. Meiotic studies of the addition lines revealed the homoeology of four B. oxyrrhina chromosomes (synteny groups 1, 3, 5 and 6 ) with B. campestris chromosomes as indicated by trivalent associations, with the highest homoeology (44.23%) in synteny group 1 and the lowest (6.1%) in synteny group 3. Seed fertility of the addition lines ranged from 94.85% (synteny group 1) to 56.98% (synteny group 5). All of the addition lines were male-sterile except synteny group 6 which had 12–16% stainable pollen. Ovule transmission of the B. oxyrrhina chromosomes added to the progenies of addition lines ranged from 23.52% (synteny group 6) to 14% (synteny group 7). RAPD analysis confirmed the validity of synteny grouping based on morphological observations. Approximately 45% of the primers studied were informative, giving B. oxyrrhina-specific RAPD bands unique for each synteny group, except group 6.
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  • 40
    ISSN: 1432-2242
    Keywords: Key words NESTUR ; Stem growth efficiency ; RAPD ; QTL ; Haploid megagametophyte
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  NESTUR (needle-to-stem unit rate) is a stem growth index of conifer seedlings that measures the efficiency of stemwood production per unit of needle growth, and is related to other seedling traits such as height, stem diameter, stem volume and needle volume. Quantitative trait loci (QTLs) affecting the expression of stem growth efficiency in radiata pine seedlings were investigated using a RAPD linkage map constructed from markers scored on haploid, megagametophytic DNA. Four putative QTLs were detected which accounted for 8.5–36.4% of the population variance. A search for evidence of epistasis, using both complete pairwise and conditional interactions, did not yield any statistically significant result. Over a 3-year period, seedlings with high-NESTUR marker alleles showed a superior growth performance of 17–40% for height, diameter and volume over those with low-NESTUR marker alleles.
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  • 41
    ISSN: 1432-2242
    Keywords: Key words Quercus robur L ; Linkage map ; RAPD ; SCAR ; Microsatellite ; Minisatellite ; 5S rDNA ; Isozymes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  A genetic map of Pedunculate oak (Quercus robur) was constructed based on one 5S rDNA, 271 RAPD, ten SCAR, 18 microsatellite, one minisatellite, and six isozyme markers. A total of 94 individuals from a full-sib family was genotyped. Two maps, including 307 markers, were constructed according to the “two-way pseudo-testcross” mapping strategy. Testcross markers segregating in the 1 : 1 ratio were first used to establish separate maternal (893.2 cM, 12 linkage groups) and paternal (921.7 cM, 12 linkage groups) maps. Both maps provided 85–90% genome coverage. Homologies between the male and female linkage groups were then identified based on 74 intercross markers segregating in the 3 : 1, 1 : 2 : 1 and 1 : 1 : 1 : 1 ratios (RAPDs, SCARs, SSRs, 5S rDNA and isozymes) in the hybrid progeny. In each map, approximately 18% of the studied markers showed segregation distortion. More than 60% of the skewed markers were due to an excess of heterozygote genotypes. This map will be used for: (1) studying the molecular organisation of genomic regions involved in inter- and intraspecific differentiation in oaks and (2) identification of QTLs for adaptive traits.
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  • 42
    ISSN: 1432-2242
    Keywords: Key words Prunus persica ; Linkage map ; RFLP ; RAPD ; AFLP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  A genetic linkage map of peach [Prunus persica (L.) Batch] was constructed in order to identify molecular markers linked to economically important agronomic traits that would be particularly useful for long-lived perennial species. An intraspecific F2 population was generated from self-pollinating a single F1 plant from a cross between a flat non-acid peach, ‘Ferjalou Jalousia®’ and an acid round nectarine ‘Fantasia’. Mendelian segregations were observed for 270 markers including four agronomic characters (peach/nectarine, flat/round fruit, acid/non-acid fruit, and pollen sterility) and 1 isoenzyme, 50 RFLP, 92 RAPD, 8 inter-microsatellite amplification (IMA), and 115 amplified fragment length polymorphism (AFLP) markers. Two hundred and forty-nine markers were mapped to 11 linkage groups covering 712 centiMorgans (cM). The average density between pairs of markers is 4.5 cM. For the four agronomic characters studied, molecular markers were identified. This map will be used for the detection of QTL controlling fruit quality in peach and, particularly, the acid and sugar content.
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  • 43
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    Theoretical and applied genetics 97 (1998), S. 968-975 
    ISSN: 1432-2242
    Keywords: Key words Triticum monococcum ; Vernalization genes ; Vrn-1 ; Vrn-2 ; RFLP ; Comparative maps
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The adaptability of Triticum aestivum to a large range of environments is partially due to genetic differences in sensitivity to vernalization. The most potent gene reducing the vernalization requirement in hexaploid wheat is Vrn-A1. An orthologous vernalization gene, designated Vrn-A m 1, was mapped in the diploid wheat Triticum monococcum between RFLP markers Xwg908 and Xabg702 on the long arm of chromosome 5AmL. The orthology of VrnA m 1 with Vrn-A1 (5A wheat, originally Vrn1), Vrn-D1 (5D wheat, originally Vrn3), Vrn-R1 (5R rye, originally Sp1) and Vrn-H1 (5H barley, originally Sh2) was shown by mapping RFLP markers linked to these vernalization genes on the T. monococcum linkage map. A second vernalization gene, designated Vrn-A m 2, was found in the distal region of chromosome 5AmL within a segment translocated from homoeologous group 4. This gene is completely linked to RFLP marker Xbcd402 and located between the same RFLP markers (Xβ-Amy-1 and Xmwg616) as the Vrn-H2 (originally Sh) locus in Hordeum vulgare.
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  • 44
    ISSN: 1432-2242
    Keywords: Key words Genetic diversity ; Triticum tauschii ; Triticum aestivum ; RFLP ; Landrace wheat
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    Notes: Abstract  Chinese accessions of Triticum tauschii and T. aestivum L. from the Sichuan white (SW), Yunnan hulled (YH), Tibetan weedrace (TW), and Xinjiang rice (XR) wheat groups were subjected to RFLP analysis. T. tauschii and landraces of T. aestivum from countries in Southwest Asia were also evaluated. For T. tauschii, a west to east gradient was apparent where the Chinese accessions exhibited less diversity than those from Southwest Asia. Compared to the Southwest Asian gene pool, the Chinese T. tauschii was highly homogeneous giving a low frequency of polymorphic bands (16%) and banding patterns (1.33 per probe) with 75 RFLP probe-HindIII combinations. Accessions of T. tauschii from Afghanistan and Pakistan were genetically more similar to the Chinese T. tauschii than those from Iran. Of 368 bands found for 39 Chinese hexaploid wheat accessions with 63 RFLP probe-HindIII combinations, 28.3% were polymorphic with an average of 2.6 banding patterns per probe and 5.0 bands per genotype. The individual Chinese landrace wheat groups revealed less variation than those from Afghanistan, Iran, and Turkey. When classified into country based groups, however, the diversity level over all Chinese landraces was greater than that of some Southwest Asian landraces, especially those from Afghanistan and Iran . The XR wheat group was genetically distinct from the other three Chinese landrace groups and was more related to the Southwest Asian landraces. The TW group was genetically similar to, but more diverse than, the SW and YH groups. The Chinese landraces had a higher degree of genetic relatedness to the Southwest Asian T. tauschii, particularly to accessions from Iran, rather than to the Chinese T. tauschii. ‘Chinese Spring’ was most related to ‘Chengdu-guang-tou’, a cultivar from the SW wheat group.
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  • 45
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    Theoretical and applied genetics 96 (1998), S. 621-627 
    ISSN: 1432-2242
    Keywords: Key words Cacao ; Theobroma cacao ; Genetic diversity ; Crop evolution ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Neotropical tree crops are affected by a combination of biological and human factors that complicate the study of genetic diversity and crop evolution. Genetic diversity and relationships among southern Mexican populations and horticultural collections of Theobroma cacao (chocolate, cocoa, cacao) are examined in light of the agricultural practices of the Maya. Collections of cacao were obtained from the extremes of its geographic range including archeological sites in southern Mexico where cacao was first domesticated. Genetic diversity was assayed by 57 informative random amplified polymorphic DNA (RAPD) marker loci. A unique sample of the total diversity found in this study exists in the southern Mexican populations. These populations are significantly different from all other cacao with regards to their profile of RAPD bands, including the ‘criollo’ variety, their morphological and geographical group. A population of cacao found in a sinkhole (cenote) in northern Yucatan with genetic affinities to populations in Chiapas suggests the Maya maintained plants far away from their native habitat. This finding concurs with known agroforestry practices of the Maya. Modern efforts to increase germplasm of tropical tree crops such as cacao should carefully examine archeological sites where genetic diversity, either deliberately or by chance, was collected and maintained by ancient cultures.
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  • 46
    ISSN: 1432-2242
    Keywords: Key words Wheat ; Aegilops comosa ; Translocation ; Homoeologous recombination ; RFLP ; C-banding ; Genomic in situ hybridization
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    Topics: Biology
    Notes: Abstract  The genetic constitutions of chromosome 2M of Aegilops comosa and the derived wheat-Ae. comosa translocations were analyzed by molecular cytogenetic techniques. Hybridization of 15 RFLP markers covering the entire length of the group-2 chromosomes revealed that chromosome 2M was structurally rearranged compared to the homoeologous chromosomes of wheat by either a pericentric inversion or a terminal intrachromosomal translocation. The breakpoint of the rearrangement was located in a region between the loci Xpsr131 and Xcdo405, resulting in the translocation of 47% of 2MS to 2ML. This aberrant structure of 2M allowed homoeologous recombination between 2M and its wheat counterpart only in the translocated segment on 2ML. C-banding and genomic in situ hybridization analyses confirmed that all translocation chromosomes consisted of the complete 2MS arm, a large part of 2ML, and very small distal segments derived from 2AS or 2DS, as expected from the aberrant structure of chromosome 2M. Thus, the translocation in the line 2A-2M?4/2 can be described as T2AS-2M?1L ⋅ 2M?1S and the translocations in the lines Compair and 2D-2M?3/8 as T2DS-2M?1L ⋅ 2M?1S. RFLP analysis determined the breakpoints in these translocation chromosomes to be within the telomeric 16% of the wheat chromosome arms. The breakpoint of the 2A/2M translocation was between Xbcd348 and Xcdo783, and that of the 2D/2M translocation was between Xcdo783 and Xpsr666. Because the translocation chromosomes retain the structural aberration found in chromosome 2M, further exploitation of the wheat-Ae. comosa translocations for cultivar improvement is questionable.
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  • 47
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    Theoretical and applied genetics 96 (1998), S. 823-831 
    ISSN: 1432-2242
    Keywords: Key words Mangifera indica L. ; Anthracnose ; Somatic embryogenesis ; Dual culture ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Embryogenic nucellar cultures of two polyembryonic mango cultivars, ‘Hindi’ and ‘Carabao’, were selected for resistance to the culture filtrate and phytotoxin of a virulent strain of Colletotrichum gloeosporioides Penz. that was isolated from mango leaves. The cultures were recurrently selected either with progressively increasing concentrations of culture filtrate or by continuous challenge with the same concentration of either culture filtrate phytotoxin. Mycelium growth was inhibited when the pathogen was cocultured with the selected, resistant embryogenic cultures. Conditioned plant growth medium containing macerated resistant embryogenic cultures did not inhibit mycelium growth, confirming that extracellular antifungal compounds were involved in the defense response. Enhanced secretion of chitinase and glucanase was observed in the plant growth medium in which resistant embryogenic cultures and regenerated somatic embryos were grown in comparison with the controls.
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  • 48
    ISSN: 1432-2242
    Keywords: Key words Citrus ; RFLP ; RAPD ; Phylogeny ; Taxonomy
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    Topics: Biology
    Notes: Abstract  Relationships among 88 accessions representing 45 Citrus species, three man-made hybrids, and six related genera were examined for restriction fragment length polymorphisms (RFLP). Thirty-two Citrus and three Microcitrus accessions were also examined by random amplified polymorphic DNA (RAPD) analysis. A measure of relative heterozygosity was estimated based on the mean of the number of fragments per individual per probe-enzyme combination (PEC) divided by total number of fragments per PEC for all non-hybrid Citrus individuals. The presence in a Citrus species of a rare band found also in a related genus was taken as an indication of possible introgression, while the presence of several fragments unique to 1 species was used to indicate non-involvement of that species in hybridization events. Most species that have been described in the literature as hybrids had high heterozygosity indices and no unique fragments. Distance matrices and dendrograms were generated using simple matching coefficient and neighbor-joining cluster analysis. RFLP and RAPD data gave approximately the same results. These data showed C. maxima was affiliated with the papedas C. hongheensis and C. latipes. C. medica clustered with C. indica when only non-hybrid taxa were examined, or among limes, lemons, and relatives when all species were considered. Mandarins did not show strongly supported groupings among themselves, nor with other species. These data showed that several accessions were probably assigned to the wrong species.
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  • 49
    ISSN: 1432-2242
    Keywords: Key words Potato virus X ; Resistance gene ; Genetic mapping ; RFLP ; Solanaceae
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    Notes: Abstract  The line IvP35 of the diploid (2n=2x=24) cultivated potato species Solanum phureja (family Solanaceae) expresses hypersensitive resistance (H) to potato X potexvirus (PVX). In this study, a diploid potato population was produced using IvP35 as the male parent and a diploid line of S. tuberosum (87HW13.7) as the female parent and tested for resistance to PVX. Data indicated that H to PVX in IvP35 is a dominant, monogenically inherited trait controlled by a single gene, named Nx phu , that is in a simplex condition (Nxnx). RFLP analysis carried out on the progeny lines revealed 4 markers (CT220, TG328, CT112 and TG424) from the long arm of chromosome IX that were linked to the hypersensitive phenotype; the closest linkage was observed with the marker TG424. Previous authors have shown that the same region of chromosome IX contains the gene Sw-5 for resistance to tomato spotted wilt tospovirus in Lycopersicon peruvianum (Solanaceae).
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  • 50
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    Theoretical and applied genetics 96 (1998), S. 844-851 
    ISSN: 1432-2242
    Keywords: Key words Common bean ; Anthracnose resistance ; RAPD ; Genetic structure ; Centre of origin
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    Notes: Abstract  The genetic structure of wild common bean populations was studied in the South-Andean centre of origin of the species. Plants were collected from 21 populations in Argentina and genetic variability was assessed for molecular and resistance markers. Polymorphism was weak for phaseolin, the major seed-storage protein, and for RAPD markers, while a high level of polymorphism was observed for resistance to anthracnose, one of the most important diseases of common bean. For the three traits, within-population variability was important and represented between 43.6% and 67.5% of the total variation. Although among-population differentiation was significant for all the traits, no correlation was found between the population distances calculated from RAPDs and resistance. These results indicate that pathogen selection pressure may be an important factor influencing the distribution of variability within and among host plant populations.
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  • 51
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    Theoretical and applied genetics 96 (1998), S. 1162-1169 
    ISSN: 1432-2242
    Keywords: Key words Wheat ; Russian wheat aphid ; Dn2 resistance gene ; RAPD ; SCAR
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    Topics: Biology
    Notes: Abstract  RAPD (random amplified polymorphic DNA) analysis was used to identify molecular markers linked to the Dn2 gene conferring resistance to the Russian wheat aphid (Diuraphis noxia Mordvilko). A set of near-isogenic lines (NILs) was screened with 300 RAPD primers for polymorphisms linked to the Dn2 gene. A total of 2700 RAPD loci were screened for linkage to the resistance locus. Four polymorphic RAPD fragments, two in coupling phase and two in repulsion phase, were identified as putative RAPD markers for the Dn2 gene. Segregation analysis of these markers in an F2 population segregating for the resistance gene revealed that all four markers were closely linked to the Dn2 locus. Linkage distances ranged from 3.3 cM to 4.4 cM. Southern analysis of the RAPD products using the cloned RAPD markers as probes confirmed the homology of the RAPD amplification products. The coupling-phase marker OPB10880c and the repulsion-phase marker OPN1400r were converted to sequence characterized amplified region (SCAR) markers. SCAR analysis of the F2 population and other resistant and susceptible South African wheat cultivars corroborated the observed linkage of the RAPD markers to the Dn2 resistance locus. These markers will be useful for marker-assisted selection of the Dn2 gene for resistance breeding and gene pyramiding.
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  • 52
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    Theoretical and applied genetics 96 (1998), S. 1151-1161 
    ISSN: 1432-2242
    Keywords: Key words European maize ; Zea mays ; Sugarcane mosaic virus ; Disease resistance ; RFLP
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    Notes: Abstract  Sugarcane mosaic virus (SCMV) causes considerable damage to maize (Zea mays L.) in Europe. The objective of the present study was to determine the genetic basis of resistance to SCMV in European maize germplasm and to compare it with that of U.S. inbred Pa405. Three resistant European inbreds D21, D32, and FAP1360A were crossed with four susceptible inbreds F7, KW1292, D408, and D145 to produce four F2 populations and three backcrosses to the susceptible parent. Screening for SCMV resistance in parental inbreds and segregating generations was done in two field trials as well as under greenhouse conditions. RFLP markers umc85, bnl6.29, umc10, umc44, and SSR marker phi075 were used in F2 populations or F3 lines to locate the resistance gene(s) in the maize genome. Segregation in the F2 and backcross generations fitted to different gene models depending on the environmental conditions and the genotype of the susceptible parent. In the field tests, resistance in the three resistant European inbreds seems to be controlled by two to three genes. Under greenhouse conditions, susceptibility to SCMV in D32 appears to be governed by one dominant and one recessive gene. Allelism tests indicated the presence of a common dominant gene (denoted as Scm1) in all three resistant European inbreds and Pa405. Marker analyses mapped two dominant genes: Scm1 on chromosome 6S and Scm2 on chromosome 3.
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  • 53
    ISSN: 1432-2242
    Keywords: Key words Wild soybean ; Glycine soja ; RFLP ; Mitochondrial DNA ; Geographic distribution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Wild soybean (Glycine soja Sieb. et Zucc.), regarded as the progenitor of cultivated soybean [G. max (L.) Merr.], is widely distributed in East Asia. We have collected 1097 G. soja plants from all over Japan and analyzed restriction fragment length polymorphisms (RFLPs) of mitochondrial DNA (mtDNA) in them. Based on the RFLPs detected by gel-blot analysis, using coxII and atp6 as probes, the collected plants were divided into 18 groups. Five mtDNA types accounted for 94% of the plants examined. The geographic distribution of mtDNA types revealed that, in many regions, wild soybeans grown in Japan consisted of a mixture of plants with different types of mtDNA, occasionally even within sites. Some of the mtDNA types showed marked geographic clines among the regions. Additionally, some wild soybeans possessed mtDNA types that were identical to those widely detected in cultivated soybeans. Our results suggest that the analysis of mtDNA could resolve the maternal lineage among plants of the genus Glycine subgenus Soja.
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  • 54
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    Theoretical and applied genetics 96 (1998), S. 203-208 
    ISSN: 1432-2242
    Keywords: Key words Genetic mapping ; Reciprocal crosses ; RFLP ; Rye ; Secale cereale L.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  A genetic linkage map of rye composed of 91 loci (88 RFLP, two morphological and one isozyme markers) has been developed using two reciprocal crosses. The RFLP loci covering all seven chromosomes were detected by a selection of rye, wheat, barley and oat cDNA and genomic DNA probes. The level of polymorphism was dependent on the source of the clones, with a ranking of rye〉wheat〉barley〉oat. Distorted segregations were detected in linkage groups of chromosomes 1R, 4R, 5R and 7R. When the recombination of the two reciprocal crosses was compared, no systematic increase or decrease in one or the other direction was observed suggesting that a combination of populations of reciprocal crosses is possible.
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  • 55
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    Theoretical and applied genetics 96 (1998), S. 361-366 
    ISSN: 1432-2242
    Keywords: Key words Centromere ; Telocentric ; Secondary trisomic ; Maize ; RFLP
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Abstract  The centromere of maize chromosome 4 was previously localized to a 26-cM interval using molecular markers and B-A translocations. The objective of the present study was to refine the placement of the centromere using secondary trisomics. Two independently isolated secondary trisomics (having an isochromosome plus two normal homologs) for 4S were recovered. RFLP analysis of populations segregating for them placed the centromere of chromosome 4 between bnl15.45 and bnl7.20, two RFLP loci that are 5.4-cM apart on the UMC map and 11.5-cM apart on the BNL map.
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  • 56
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    Theoretical and applied genetics 96 (1998), S. 354-360 
    ISSN: 1432-2242
    Keywords: Key words Soybean ; Glycine max ; QTL ; RFLP ; SLW ; Leaf size
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    Notes: Abstract  Selection for high specific leaf weight (SLW) in soybean [Glycine max (L) Merr.] may increase apparent photosynthetic rate per unit leaf area (AP), which in turn may improve seed yield. In general, the SLW and leaf size are negatively correlated in soybean. To maximize total photosynthetic performance, and perhaps the seed yield, of a soybean cultivar, it would be necessary to establish a large leaf area rapidly while maintaining a high SLW. The objective of the present study was to identify quantitative trait loci (QTLs) conditioning SLW and leaf size in soybean. One hundred and twenty F4-derived lines from a ‘Young’×PI416937 population were evaluated using restriction fragment length polymorphism (RFLP) markers. The genetic map consisted of 155 loci on 33 linkage groups (LGs) covering 973 cM of map distance. The phenotypic data were collected from two different environments – a greenhouse at Athens, Ga. and a field site at Windblow, N.C. The SLW and leaf-size measurements were made on leaves from the 8th and 9th node of soybean plants at the V12 stage of development. Combined over environments, six putative independent RFLP markers were associated with SLW, and four of these loci were consistent across environments. Individually, the six markers each explained between 8 and 18% of the phenotypic variation among lines for SLW. The Young alleles contributed to a greater SLW at four of the six independent marker loci, and transgressive segregation occurred among the progeny for SLW. Three putative independent RFLP markers were associated with leaf size, each explaining between 6 to 11% of the phenotypic variation in the trait, and one of these markers was identified in both environments. There was no correlation between SLW and leaf size in this population. Similarly, none of the six QTLs conditioning SLW were linked to any of the three QTLs for leaf size. In this soybean population, it is possible to select for progeny lines with greater SLW than either parent perhaps without affecting the leaf size. It is feasible to pyramid all of the desirable alleles for greater SLW and large leaf size in a single genetic background.
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    Theoretical and applied genetics 97 (1998), S. 83-89 
    ISSN: 1432-2242
    Keywords: Key words Lens culinaris subsp. orientalis ; Recombinant inbred lines ; AFLP ; RAPD ; Genetic map
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  A genetic linkage map of Lens sp. was constructed with 177 markers (89 RAPD, 79 AFLP, six RFLP and three morphological markers) using 86 recombinant inbred lines (F6:8) obtained from a partially interspecific cross. The map covered 1073 cM of the lentil genome with an average distance of 6.0 cM between adjacent markers. Previously mapped RFLP markers were used as anchor probes. The morphological markers, pod indehiscence, seed-coat pattern and flower-color loci were mapped. Out of the total linked loci, 8.4% showed segregation distortion. More than one-fourth of the distorted loci were clustered in one linkage group. AFLP markers showed more segregation distortion than the RAPD markers. The AFLP and RAPD markers were intermingled and clustering of AFLPs was seldom observed. This is the most extensive genetic linkage map of lentil to-date. The marker density of this map could be used for the identification of markers linked to quantitative trait loci in this population.
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  • 58
    ISSN: 1432-2242
    Keywords: Key words Zea mays L ; AFLP ; RFLP ; Testcross mean ; Testcross variance ; Genetic distance ; Midparent heterosis
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    Notes: Abstract  Prediction of the means and genetic variances in segregating generations could help to assess the breeding potential of base populations. In this study, we investigated whether the testcross (TC) means and variances of F3 progenies from F1 crosses in European maize can be predicted from the TC means of their parents and F1 crosses and four measures of parental genetic divergence: genetic distance (GD) determined by 194 RFLP or 691 AFLPTM 1 markers, mid-parent heterosis (MPH), and absolute difference between the TC means of parents (∣P1−P2∣). The experimental materials comprised six sets of crosses; each set consisted of four elite inbreds from the flint or dent germplasm and the six possible F1 crosses between them, which were evaluated for mid-parent heterosis. Testcross progenies of these materials and 20 random F3 plants per F1 cross were produced with a single-cross tester from the opposite heterotic group and evaluated in two environments. The characters studied were plant height, dry matter content and grain yield. The genetic distance between parent lines ranged between 0.17 and 0.70 for RFLPs and between 0.14 and 0.57 for AFLPs in the six sets. Testcross-means of parents, F1 crosses, and F3 populations averaged across the six crosses in a particular set generally agreed well for all three traits. Bartlett’s test revealed heterogeneous TC variances among the six crosses in all sets for plant height, in four sets for grain yield and in five sets for dry matter content. Correlations among the TC means of the parents, F1 crosses, and F3 populations were highly significant and positive for all traits. Estimates of the TC variance among F3 progenies for the 36 crosses showed only low correlations with the four measures of parental genetic divergence for all traits. The results demonstrated that for our material, the TC means of the parents or the parental F1 cross can be used as predictors for the TC means of F3 populations. However, the prediction of the TC variance remains an unsolved problem.
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    Theoretical and applied genetics 97 (1998), S. 99-102 
    ISSN: 1432-2242
    Keywords: Key words Genetic mapping ; CMS ; Fertility restoration ; RAPD ; RFLP ; Rye ; Secale cereale L.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  A gene determining the restoration of cytoplasmic genic male sterility (CMS) caused by the Gülzow (G)-type cytoplasm was mapped by analyzing an F2 and F3 population comprising 140 and 133 individual plants, respectively. The target gene, designated Rfg1, was mapped on chromosome 4RL distally to three RFLP (Xpsr119, Xpsr167, Xpsr899) and four RAPD (XP01, XAP05, XR11, XS10) loci. Xpsr167 and Xpsr899 are known to be located on the segment of chromosome 4RL which was ancestrally translocated and is homoeologous to the distal end of other Triticeae 6S chromosomes. It is suggested that Rfg1 may be allelic to the gene determining the restoration of rye CMS caused by the Pampa (P) cytoplasm (chromosome 4RL) and to Rfc4 that on rye addition lines of chromosome 4RL restores male fertility of hexaploid wheat with T. timopheevi cytoplasm. Homoeoallelism to two loci for cytoplasmic-male-sterility restoration on chromosomes 6AS and 6BS in hexaploid wheat is also suggested.
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    Theoretical and applied genetics 97 (1998), S. 327-336 
    ISSN: 1432-2242
    Keywords: Key words STS ; Codominant PCR marker ; RFLP ; Loblolly pine (Pinus taeda L.) ; Comparative map
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    Topics: Biology
    Notes: Abstract  We report a strategy for developing codominant PCR-based genetic markers by using sequenced cDNA clones from loblolly pine (Pinus taeda L.). These clones were previously used as probes for detecting restriction fragment length polymorphisms (RFLPs) to generate linkage maps. After assessing the complexity of banding patterns from Southern blots, we selected clones representing relatively simple gene families, and then determined nucleotide sequences for about 200 bp at each end of the cDNA inserts. Specific PCR primers were designed to amplify samples of genomic DNA derived from two loblolly pine mapping populations. Polymorphisms were detected after digesting the amplified DNA fragments with a battery of restriction endonucleases, and most polymorphisms were inherited in a Mendelian fashion. These newly identified genetic markers are codominant and relatively simple to use. By assaying DNA from individuals used to construct RFLP maps, we show that most of these markers map to the same position as the RFLP loci detected using their corresponding cDNAs as probes, implying that these markers have been converted from RFLP to PCR-based methods. These PCR-based markers will be useful for genome mapping and population genetics.
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  • 61
    ISSN: 1432-2242
    Keywords: Key words Targeted mapping ; RFLP ; RAPD ; Brassica napus ; Polima CMS ; Nearly isogenic line ; Bulked segregant analysis
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    Notes: Abstract  We have used two targeting approaches [pairs of nearly isogenic lines (NILs) and bulked segregant analysis] to identify DNA markers linked to the Rfp1 restorer gene for the pol CMS of canola (Brassica napus L.). We were able to target the Rfp1 locus as efficiently by comparing NILs as by bulked segregant analysis, and it was demonstrated in this instance that double-screening strategies could significantly improve the overall targeting efficiency. The chance occurrence of shared homozygosity at specific unlinked chromosomal regions in the bulks was found to limit the efficiency of bulked segregant analysis, while the efficiency of NIL comparison was limited by residual DNA from the donor cultivar at scattered sites throughout the genome of the NILs.
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    Theoretical and applied genetics 97 (1998), S. 422-430 
    ISSN: 1432-2242
    Keywords: Key words Sunflower ; Helianthus ; Polyploidy ; Genomes ; RAPD
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    Topics: Biology
    Notes: Abstract  Forty taxa belonging to 36 species and four unclassified accessions of Helianthus were studied using RAPD technology. Single ten-mer primers were screened for those amplifying fragments common to several species. We found that when several species shared a common fragment, they belong to the same section of the genus. Moreover, we also found that some fragments are common to all species of the Helianthus. Most of the fragments were found to be of the same size in these species and to share the homology indicated by molecular hybridization. Out of 118 retained fragments, 33 were common to all Helianthus species, 56 were unique to perennial species of sects. Atrorubentes and Ciliares, 24 were unique to sect. Atrorubentes, 29 were unique to sect. Helianthus, whereas 0 were unique to sect. Ciliares. Each set of common or specific fragments was assumed to belong to a genome: (1) the C genome carrying the fragments common to all species of the three sections, (2) the H genome unique to sect. Helianthus, (3) the P genome common to perennial species (sects. Atrorubentes and Ciliares), and (4) the A genome unique to sect. Atrorubentes. The genomic structure was therefore HC for sect. Helianthus, CPA for sect. Atrorubentes, and CP? for sect. Ciliares. Molecular hybridizations with amplification products revealed homologies between Helianthus genomes and several other genera in the Helianthinae sub-tribe. The simple method used to characterize these fragments led to powerful tools for recognizing genomes which reconcile the section organization of the genus and the degree of difficulty in crossing perennial and annual forms.
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    Theoretical and applied genetics 97 (1998), S. 439-445 
    ISSN: 1432-2242
    Keywords: Key words Actinidia ; RAPD ; SCAR ; Sex-linked markers ; Marker-assisted selection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Two sex-linked random amplified polymorphic DNA (RAPD) markers identified from Actinidia chinensis were converted into sequence-characterised amplified regions (SCARs) for the large-scale screening of Actinidia breeding populations. Initial SCAR primers converted one RAPD (SmX) into a dominant marker, but the other (SmY), which was potentially more useful because of its linkage to the male determining ‘Y’ locus, failed to retain polymorphism. This difficulty was overcome by cloning and sequencing the alternate ‘allele’ from female plants, and then designing ‘allele’-specific primers that utilised nucleotide differences between the sexes. Using a quick squash-blot method of DNA extraction, the SCAR primers were tested in 120 A. chinensis plants to determine their gender. The system is now in use for large-scale screening of seedling populations in the Actinidia breeding programme. The sex-linked SCAR primers also functioned with plants from some other geographically separate accessions of A. chinensis and with plants in the closely related polyploid species A. deliciosa, but did not amplify a sex-linked band in more distantly related species of Actinidia.
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  • 64
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    Theoretical and applied genetics 97 (1998), S. 657-670 
    ISSN: 1432-2242
    Keywords: Key words Triticum aestivum ; Phylogeny ; Genetic distance ; Genome ; Introgression ; Allopolyploidy ; RFLP ; Glutenin ; rRNA ; Non-transcribed spacers ; Evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Polymorphism in the lengths of restriction fragments at 53 single-copy loci, the rRNA locus Nor3, and the high-molecular-weight glutenin locus Glu1 was investigated in the D genome of hexaploid Triticum aestivum and that of Aegilops tauschii, the source of the T. aestivum D genome. The distribution of genetic variation in Ae. tauschii suggests gene flow between Ae. tauschii ssp. strangulata and ssp. tauschii in Iran but less in Transcaucasia. The “strangulata” genepool is wider than it appears on the basis of morphology and includes ssp. strangulata in Transcaucasia and southeastern (SE) Caspian Iran and ssp. tauschii in north-central Iran and southwestern (SW) Caspian Iran. In the latter region, Ae. tauschii morphological varieties ‘meyeri’ and ‘typica’ are equidistant to ssp. strangulata in Transcaucasia, and both belong to the “strangulata” genepool. A model of the evolution of Ae. tauschii is presented. On the geographic region basis, the D genomes of all investigated forms of T. aestivum are most closely related to the “strangulata” genepool in Transcaucasia, Armenia in particular, and SW Caspian Iran. It is suggested that the principal area of the origin of T. aestivum is Armenia, but the SW coastal area of the Caspian Sea and a corridor between the two areas may have played a role as well. Little genetic differentiation was found among the D genomes of all investigated free-threshing and hulled forms of T. aestivum, and all appear to share a single D-genome genepool, in spite of the fact that several Ae. tauschii parents were involved in the evolution of T. aestivum.
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  • 65
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    Theoretical and applied genetics 97 (1998), S. 1042-1046 
    ISSN: 1432-2242
    Keywords: Key words T. aestivum ; H. villosa ; C-banding ; RFLP ; Deletion mapping ; Pm21
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Three deletion lines (del6V?2S-1, del6V? 2L-1, and del6V?2L-2) of Haynaldia villosa chromosome 6V added to wheat were identified by C-banding and characterized by RFLP analyses. The breakpoints were located at fraction lengths (FL) 0.58 in del6V?2S-1 in the short arm, and FL 0.66 in del6V?2L-1 and FL 0.64 in del6V?2L-2 in the long arm. Thirty-one Triticeae homoeologous group-6 DNA probes were used to map RFLP loci in the deletion lines and the wheat-H. villosa disomic substitution (DS) line 6V?2(6A). Nine probes failed to detect polymorphism between Chinese Spring and DS6V?2(6A). Ten of sixteen polymorphic short-arm loci were not detected in del6V?2S-1. Thus, the loci are located in the deleted distal chromosome region. Six RFLP markers were mapped in the proximal 58% of 6VS. Of 20 DNA markers specific for 6VL, six mapped in the distal 36% of the long arm, and nine mapped in the proximal 64% of 6VL. The breakpoint of the short arm of 6V?2 occurs between Xpsr106 and Xcdo270, and that of the long arm between Xpsr915 and Xmwg934. The powdery mildew resistance gene Pm21 is located on the short arm of chromosome 6V?2. Pm21 is present in del6V?2S-1, and can be further mapped in the proximal 58% of 6V?2S.
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  • 66
    ISSN: 1618-2545
    Keywords: intraspecific group ; isozymes ; morphology ; Pythium ultimum var.ultimum ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Comparisons were made between two morphological groups ofPythium ultimum var.ultimum strains isolated in a vegetable field in Japan. The groups were distinguished as having smaller or larger sexual organs by the sizes of their antheridia and oogonia. Morphological study indicated that the two groups comprised a single taxon,P. ultimum var.ultimum, by the current taxonomical keys. The smaller group grew faster in the lower temperature range of 4–15°C, whereas the larger group grew faster in the higher temperature range of 25–37°C. Random amplified polymorphic DNA (RAPD) and isozyme analyses revealed genetic dissimilarity between the two groups. Cluster analysis of the isozyme banding patterns with four otherPythium spp. demonstrated that the genetic dissimilarity between the two groups was equivalent to species level. In the field survey, the smaller group was frequently detected in February, May and September but not in July, while the larger group was detected mainly in July and September. The two groups were not distinguishable by their pathogenicity to spinach seedlings.
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  • 67
    ISSN: 1618-2545
    Keywords: Coprinus ; RAPD ; strain identification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract All five examined strains ofCoprinus cinereus could be clearly discriminated from the strains of five otherCoprinus species by RAPD patterns with 12 of 13 primers. Also one specimen of unknownCoprinus strain was identified to beC. cinereus by this method. The RAPD patterns were similar among the strains in the same species; many common DNA fragments were recognized as well as some strain-specific DNA fragments. Thus all seven strains ofC. cinereus and all four strains ofC. angulatus examined could be distinguished individually. Diakryotic strains showed the combined RAPD patterns of the two monokaryotic strains constituting the dikaryon. The combined RAPD markers observed in the dikaryons were segregated in their basidiospore progeny. All 18 randomly picked progeny showed different combinations of RAPD markers from the parental strains.
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  • 68
    ISSN: 1573-5176
    Keywords: Dunaliella ; ITS ; PCR ; RFLP ; strains ; taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The genus Dunaliella comprises 28 species defined primarily by morphological and physiological criteria, which vary considerably depending on growth conditions. Concomitantly, the taxonomic status of various species is uncertain. To confirm the taxonomic identity and to better understand the relationship within Dunaliella, seven taxa ( D. salina, D. bardawil, D. tertiolecta, D. parva, D. viridis, D. lateralis, D. peircei) were compared using RFLP analysis of the nuclear rDNA repeats, specifically the internal transcribed spacer regions, including the 5.8S rRNA gene. Volvox aureus was used as an outgroup. A single ITS PCR amplification product was obtained for each taxon. An ITS fragment of ca. 640 bp was present in all the taxa within the subgenus Dunaliella, except for D. salina CCMP 1303 (ca. 540 bp) and D. lateralis (subgenus Pascheria) (ca. 600 bp). A cluster analysis based on the presence or absence of bands generated by digestion of the PCR product with 8 restriction endonucleases (DpnI, HhaI, EcoRI, PvuII, TaqI, HaeIII, MspI, StyI) revealed no correlation between the genetic relationship inferred from the ITS-RFLP data and the morpho-physiological attributes used for taxonomy. In addition, differences in morphology, physiology and in the length and restriction fragment patterns of the ITS region of D. salina CCMP 1303 suggest that this strain does not belong to Dunaliella.
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  • 69
    ISSN: 1572-9788
    Keywords: transgenic rice ; particle bombardment ; cell electroporation ; RAPD ; AFLP ; AFRP ; RAMP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract In the present work we utilised some of the most discriminative molecular tools, such as RAPD, AFLP, AFRP and RAMP, to analyse the genome of independently derived transgenic plants from three elite Italian cultivars (cv. Lido, Carnaroli and Thaibonnet) and found that two methods for direct gene transfer, namely particle bombardment and intact cell electroporation (the latter being a procedure set up in this work), result in transgenic rice (Oryza sativa L.) plants that exhibit negligible genomic changes. This is in contrast with recently published results showing relevant changes in the DNA of transgenic rice plants generated through protoplasts electroporation and of transgenic poplar plants engineered through Agrobacterium tumefaciens infection. Implications of these findings are discussed in the context of selecting appropriate gene transfer methodologies to produce transgenic plants expressing genes of interest while retaining their genomic integrity and, thus, their superior agronomic and/or industrial traits.
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  • 70
    ISSN: 1572-9788
    Keywords: Helianthus annuus ; QTL ; resistance ; RFLP ; Sclerotinia scerotorum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Restriction fragment length polymorphism and isoenzyme markers were used to investigate quantitative trait loci involved in sunflower resistance to mycelial extension of Sclerotinia sclerotiorum on leaves and capitula. Seed weight, oil content and flowering data were also evaluated. Four quantitative trait loci were demonstrated for leaf resistance and two for capitulum resistance. One of these zones appears involved in resistance to both types of S. sclerotiorum attack while the others appear specific for resistance of one part of the plant. Two quantitative trait loci were detected for seed weight, three for oil content and three for flowering date. Individual quantitative trait loci explained 9% to 48% of the phenotypic variability, confirming the polygenic basis of the quantitative traits studied. Overall, the quantitative trait loci explain 60% of the genetic variation for leaf resistance and 38% for capitulum resistance to S. sclerotiorum. One linkage group is particularly interesting since it includes quantitative trait loci for all the five quantitative traits measured. Hypotheses for linkage versus pleiotropy and consequences of all the results in resistance breeding are discussed.
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  • 71
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    Molecular breeding 4 (1998), S. 227-234 
    ISSN: 1572-9788
    Keywords: Helianthus annuus ; PCR ; Puccinia helianthi ; RAPD ; rust resistance ; sunflower
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract In this study we report on the identification of molecular markers, OX20600 and OO04950, linked to the geneR Adv in the proprietary inbred line P2. This gene confers resistance to most of the pathotypes of Puccinia helianthi identified in Australia. Analysis indicates these RAPD markers are linked to the resistance locus at 0.0 cM and 11 cM respectively. SCAR markers SCX20600 and SCO04950 derived from these two RAPD markers, and SCT06950 derived from a previously reported RAPD marker linked at 4.5 cM from the R 1 rust resistance gene were developed. SCX20600 and SCO04950 were linked at similar distances from their resistance locus as the RAPD markers. SCTO6950 co-segregated completely with rust resistance. The robustness of the R 1 SCAR marker was demonstrated through the amplification of the marker in a diverse range of sunflower germplasm considered to possess the R 1 gene. The SCAR markers forR Adv were not amplified in the sunflower rust differential set thereby supporting the contention that this is a novel resistance gene. They did amplify in a number of proprietary lines closely related to the line P2. This locus is under further investigation as it will be useful in our attempts to use molecular-assisted breeding to produce durable resistance in sunflower to P. helianthi.
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  • 72
    ISSN: 1572-9788
    Keywords: composite interval mapping ; maize streak virus ; QTL ; RFLP ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Resistance to maize streak virus (MSV) is an essential trait of improved maize varieties in sub-Saharan Africa. We mapped quantitative trait loci (QTL) for resistance to MSV in a population of 196 F2:3 lines derived from a cross between the maize inbred lines CML202 (resistant) from CIMMYT-Zimbabwe and Lo951 (susceptible) from Italy. Field tests were planted at two locations in Zimbabwe, inoculated with viruliferous leaf hoppers (Cicadulina mbila), and scored twice (21 and 83 days after infesting, DAI) on a 1–5 scale. The mean final streak intensity (score 2) of the parent lines was 2.2 (CML202) and 4.8 (Lo951). Genotype × location interaction was large for score 1 but negligible for score 2. Consequently, the heritability was higher for score 2 (0.93) than for score 1 (0.62). By composite interval mapping across locations, using a linkage map with 110 RFLP loci, four significant (LOD ≥3.0) QTL were identified for score 1 on chromosomes (C) 1, 2, 3, and 4, respectively. All four were contributed by CML202. For score 2, only the QTL on C 1 was significant (LOD =37), explaining 59% of the phenotypic and 64% of the genotypic variance. The QTL's partially dominant gene action was consistent with the nearly intermediate resistance of the F1 generation (relative heterosis for resistance 12%). The presence of one major QTL is consistent with the bimodal frequency distribution of the mapping population showing a clear 3:1 segregation. This gene seems to be allelic or identical to Msv1, a major resistance gene which was previously identified in the same genomic region in Tzi4, an inbred line from IITA. Inbred CML202 had lower final disease ratings than Tzi4. The greater resistance of CML202 may be due to allelic differences at the msv1 locus or due to the minor QTL on C 2, 3, and 4 which were not detected in Tzi4. z y Trigo (International Maize and Wheat Improvement Center); IITA, International Institute of Tropical Agriculture; IRAT, Institute de Recherches Agronomiques Tropicales et des Cultures Vivrières; KARI, Kenya Agricultural Research Institute; MSV, maize streak virus; QTL, quantitative trait locus/loci
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  • 73
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    European journal of plant pathology 104 (1998), S. 611-617 
    ISSN: 1573-8469
    Keywords: Exserohilum turcicum ; Johnson grass ; maize ; northern corn leaf blight ; population genetic structure ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Setosphaeria turcica is the causal agent of northern corn leaf blight, a foliar maize disease of worldwide economic importance. In Europe, its severity increases. To investigate the pathogen's population-genetic structure in central Europe, a total of 80 isolates was sampled in Germany, Switzerland, France, Austria, and Hungary and investigated with 52 random amplified polymorphic DNA (RAPD) markers. The mating type of the isolates was determined in testcrosses. Among the 73 isolates from maize there were 26 different RAPD haplotypes. All isolates with identical haplotype are considered clonemates. The haplotype shared by most members was represented by 22 isolates from Germany, Switzerland, and France, indicating high fitness and substantial migration. Only a single clone had members in both southeastern Austria and southwestern Switzerland, suggesting that the Alps constitute a major barrier for this pathogen. Several haplotypes differed by only one or two RAPD bands from the predominant haplotype and may have arisen by mutation. Few other clonal lineages were detected. The evolution of some haplotypes could not be explained by mutation alone. Sexual recombination may rarely occur. In population samples from Germany, Switzerland, and France, mating type MAT2 was predominating, while most isolates from Austria and Hungary had MAT1. Seven isolates from Johnson grass (Sorghum halepense), an alternative host of S. turcica, were clonemates and very different in RAPD haplotypes from all isolates collected from maize.
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  • 74
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    Plant systematics and evolution 212 (1998), S. 53-77 
    ISSN: 1615-6110
    Keywords: Leguminosae ; Leucaena leucocephala ; L. diversifolia ; L. ×spontanea ; Hybridization ; hybrid detection ; spontaneous hybrid ; RAPD ; RFLP ; chloroplast DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The detection of hybridity inLeucaena is discussed in relation to: (i) traditional criteria, (ii) molecular criteria and (iii) models to predict hybrid leaf morphology. Morphological, geographical and molecular evidence for the occurrence of interspecific hybrids betweenL. leucocephala andL. diversifolia in south-central Mexico, northern Guatemala, Jamaica, Dominican Republic, the Philippines and Papua New Guinea is presented. Predicted mean hybrid leaf trait values calculated from parent material are compared with data from putative hybrids and shown to be similar. The origin of these hybrids is discussed and shown to be the result of artificial sympatry resulting from indigenous, and recent exotic, domestication of the parent species. The hybrid is described asL. ×spontanea.
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  • 75
    ISSN: 1615-6110
    Keywords: Leguminosae ; Phaseolus ; Isozymes ; genetic distance ; Lima bean ; rDNA ; RFLP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Genetic variation inPhaseolus lunatus (Lima bean) was investigated at isozyme and DNA levels. Sixty cultivated accessions, including representatives of the Mesoamerican and Andean gene pools and intermediate types, were analyzed for variability at 17 isozyme loci. Some accessions were also examined for restriction fragment length polymorphism (RFLP) at the rDNA level. These data were used to construct two dendrograms showing clear separation in two distinct groups corresponding to each of the gene pools and an intermediate one probably representing a transitional group.
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  • 76
    ISSN: 1573-5036
    Keywords: chalcone isomerase ; chalcone synthetase ; chitinase ; PAL ; phaseolin ; Phaseolus vulgaris ; QTL ; RFLP ; Rhizobium tropici ; Xanthomonas campestris bv. phaseoli
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A recently developed bean RFLP linkage map was used to identify genetic elements affecting quantitative trait loci (QTLs) in two contrasting common bean genotypes, BAT-93 and Jalo EEP558, under two levels of mineral nitrogen: low – 0.25 mM NH4NO3 and a high – 6 mM NH4NO3. QTLs affecting nodule number (NN) and response to Xanthomonas campestris bv. phaseoli, which causes common bacterial blight (CBB) were identified and mapped. Analyses of 70 F2-derived F3 families, using the F1, the two parents, and a nodulation-defective mutant (Nod-) inoculated with R. tropici UM1899 under both levels of N showed significant differences (P#60;0.0001) among the F3 families for NN. Under low N, three genomic regions influenced both traits, with seven linked markers. In three of the six regions influencing NN, higher NN was associated with the Jalo EEP-558 allele, whereas in only two regions was the BAT-93 allele associated with higher NN. One-way analysis of variance, with each marker as the independent variable and NN as the dependent variable, and interval mapping analysis identified four QTLs, which accounted for 45% of the total variation, and two additional QTLs near to yet unassigned loci. In linkage group D7, one QTL mapped to the same region as a QTL for CBB. Under high N, three additional regions were linked to NN, one where the BAT-93 allele was closely associated with CH18 (chitinase), and the others where the Jalo EEP-558 allele was associated with CHS (chalcone synthetase) and PAL-1 (phenylalanine ammonia lyase). Four regions for CBB were mapped adjacent to or in the same region as a QTL for NN. Thus, N showed dual and opposite effects on the expression of NN and CBB. Analysis of these RFLP markers revealed these ‘hidden’ favorable alleles and can serve as an indirect selection tool to increase NN and resistance to CBB.
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  • 77
    ISSN: 1573-5060
    Keywords: Cocos nucifera ; RFLP ; genetic diversity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A study of the genetic diversity in coconut by RFLP analysis was performed in 100 individuals representing 10 Tall and seven Dwarf local populations or 'ecotypes' from various geographical origins. Nine cDNA clones from rice, one mitochondrial DNA clone (CoxI) and one genomic clone (rDNA) from wheat were used as probe for southern hybridization. The distribution of the 40 polymorphic bands revealed by rice cDNA clones was studied using a multivariate analysis and allowed to identify two main genetical groups. The first one includes the ecotypes from the Far East and from the South Pacific, whereas the other one comprises the ecotypes from India, Sri Lanka and Western Africa. The rDNA and the CoxI probes confirm this distinction. The Far East and the Pacific regions which were the most likely center of origin also exhibit the widest polymorphism. The associations between the Panama Tall and the Pacific group and between the West African Tall and the Indian Ocean group reflect their likely origin. The Comoro Tall appears to be intermediate between the two main groups, and could reflect the old migration route between Indonesia and Madagascar. All Dwarf varieties belong to the first group, even those collected in West Africa. Those were probably introduced from Asia and Pacific at the beginning of the century. The cross-pollinating Tall ecotypes were generally more polymorphic than the self pollinating Dwarf ecotypes. The legitimacy of two hybrids between ecotypes was confirmed and maternal inheritance of mitochondrial genome was observed.
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  • 78
    ISSN: 1573-5060
    Keywords: sweet potato ; Ipomoea batatas ; RAPD ; genetic variation ; Chile
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract RAPD (randomly amplified polymorphic DNA) technology was applied to analyze the genetic variability of sweet potato germplasm existing in Chile and elsewhere. Analysis of 28 cultivars from all over the world showed polymorphic bands with all 18 primers tested. A total of 124 RAPD bands were scored with an average of 6.9 polymorphic bands per primer. These results confirm that sweet potato exhibits high genetic variation. Two groups were distinguished: one containing Peruvian cultivars, and another containing cultivars from the rest of the world. Analysis of 14 accessions from Central Chile and one from Northern Chile showed polymorphic bands with 24 of 26 primers tested, but almost all of the 140 polymorphic bands merely showed the distinctness of the Northern accession. The almost complete uniformity of the other 14 accessions shows that sweet potato germplasm collected in Central Chile has very little genetic variability and may be derived from a single cultivar. Based on these results and on historical records, some hypotheses are proposed to explain the origin of sweet potatoes cultivated in Chile.
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  • 79
    ISSN: 1573-5060
    Keywords: cultivar identification ; fingerprinting ; genetic similarity ; Juglans regia ; RAPD ; walnut
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The potential use of the Randomly Amplified Polymorphic DNA (RAPD) technique for characterization and assessment of genetic relationships was investigated in nineteen walnut (Juglans regia L.) genotypes used as parents or released as cultivars from the breeding program of the University of California at Davis. Most of the 72 decamer primers used yielded scorable amplification patterns based on discernable bands. The results obtained produced a unique fingerprint for each of the walnut genotypes studied. Cluster analysis separated the 19 walnut genotypes into two main groups whose differences were related to their pedigree. Genotypes sharing common parents tend to group together and with at least one of the parents. Thus, RAPD markers can detect enough polymorphism to differentiate among walnut genotypes, even among closely related genotypes, and the genetic similarity based on RAPDs appears to reflect the known pedigree information. RAPD technology can be useful in current walnut breeding programs, allowing the identification of new cultivars as well as the assessment of the genetic similarity among genotypes which will help in selecting the best parents to obtain new genetic combinations.
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  • 80
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    Euphytica 101 (1998), S. 249-255 
    ISSN: 1573-5060
    Keywords: avocado ; classification ; ecological races ; Persea americana ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract RAPD (randomly amplified polymorphic DNA) analysis was carried out on 16 accessions representing the three ecological races of avocado (Persea americana Mill.), and one accession of P. schiedeana Nees. Twenty two preselected primers produced 133 polymorphic DNA fragments in the RAPD assay of the avocado accessions. One primer was identified which could differentiate each of the avocado accessions. Potentially race-specific markers for each of the Mexican, Guatemalan, and West Indian races, have been detected. A Jaccard's similarity coefficient matrix was generated and a dendrogram constructed using UPGMA (unweighted pair-group method of arithmetic averages) cluster analysis. Percentage similarity between avocado accessions ranged from 46% to 85%. The lowest similarity (between 22% and 29%) was revealed between P. schiedeana and any P. americana accession. Average similarity within races of avocado was 75% for the Mexican race, 71% for the West Indian race and 73% for the Guatemalan race. Average similarity between races ranged from 53% to 58%. The dendrogram identified three groups, representing the races of avocado. These results are in concordance with the present classification of avocado into three subspecies (varieties) of P. americana, namely drymifolia, americana, and guatemalensis, corresponding to the Mexican, West Indian and Guatemalan races, respectively, and confirm the separate species status of P. schiedeana. We conclude that RAPD markers may be useful for the classification of avocado and for the assessment of genetic diversity of avocado germplasm.
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  • 81
    ISSN: 1573-5060
    Keywords: genetic diversity ; linkage drag ; RFLP ; Stenodiplosis sorghicola ; sorghum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract In recent years, hybrids with levels of resistance to sorghum midge (Stenodiplosis sorghicola Coquillett) have become available to Australian sorghum producers. These hybrids have been readily accepted to the extent that more than 80% of the sorghum growing area was planted to hybrids with some level of midge resistance by 1995. Since selection for resistance to sorghum midge is one of the primary objectives of Australian sorghum breeding programs, the relationship between resistance and genetic diversity was investigated. Genetic diversity and heterozygosity were assessed using restriction fragment length polymorphism analysis among 26 grain sorghum hybrids grown commercially in Australia. The genetic distances between each sorghum hybrid and a standard highly resistant hybrid were found to be strongly negatively correlated to hybrid midge resistance ratings (r = - 0.77, p 〈 0.001). In addition, the average heterozygosity of each hybrid was correlated with their midge resistance ratings (r = - 0.54, p 〈 0.01). The results indicate that the move to midge resistant hybrids has been associated with a narrowing of the genetic diversity and average heterozygosity of commercial sorghum hybrids. Repeated use of particular elite parent lines, linkage drag and genetic drift are likely to have contributed to this decline. This reduction in genetic diversity may have implications for the genetic vulnerability of sorghum in Australia and the rate of progress in breeding for yield.
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  • 82
    ISSN: 1573-5060
    Keywords: Brassica napus ; RAPD ; bulked DNA ; DNA fingerprinting
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Since DNA-based markers are unaffected by environmental or physiological factors, they have potential utility in the description of plant cultivars required for award of proprietary rights (i.e. Plant Breeders' Rights). The high discriminating power of this class of markers, however, can also make demonstration of uniformity and stability of such a marker within a cultivar difficult, especially for genetically-complex cultivars. This report examines the usefulness of bulking equal quantities of DNA from 14 to 20 individuals of a cultivar to identification of RAPD DNA markers that distinguish between Brassica napus cultivars of varying genetic complexity. For the four cultivars assessed (Quantum, OAC Springfield, Innovator and AC Excel), it is shown that consistent presence/absence scores are obtained from bulked DNA samples for three different RAPD markers despite a significant degree of variation among samples from individuals. Use of bulked DNA samples thus may enable identification of a distinguishing profile of RAPD markers whose presence/absence is uniform and stable even in complex cultivars. Nevertheless, RAPD markers remain limited in that they are not strictly quantitative in nature. This limitation is discussed with respect to cultivar description for plant breeders' rights applications.
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  • 83
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    Euphytica 104 (1998), S. 181-189 
    ISSN: 1573-5060
    Keywords: Genetic diversity ; macha wheat ; spelta wheat ; RAPD ; accession duplication
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Genetic diversity in a crop species is basic to improvement of the species and can be estimated at the molecular level. The objective of this study was to estimate genetic diversity within and between spelta and macha wheats. Random amplified polymorphic DNA (RAPD) analysis was conducted on 69 spelta and 32 macha wheat accessions. The classification of spelta and macha accessions, based on Jaccard genetic similarity coefficients for RAPD markers, was consistent with their geographic origin. The results indicated that the germplasm of macha wheat was more diverse than that of spelta wheat. In the dendrogram of macha wheat, four spelta-like accessions grouped together, separate from the remaining macha accessions, suggesting that these accessions were misclassified. In addition, accessions with identical RAPD patterns were found, indicating that these accessions were probably duplicated. Thus RAPD analysis can be used to estimate genetic diversity and identify duplicate accessions in wheat germplasm collections.
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  • 84
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    Euphytica 99 (1998), S. 167-173 
    ISSN: 1573-5060
    Keywords: AFLP ; Capsicum annuum ; genetic distances ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Genetic relationships were examined among thirty-four pepper (Capsicum annuum) cultivars of different types. Two types of PCR-based markers were used, RAPD and AFLP, and their relative effectiveness was compared. A dendrogram based on RAPD markers separated the large-fruited sweet cultivars from the small-fruited pungent peppers, and the former group showed less divergence than the latter. The percentage of polymorphic markers was lower for AFLP than for RAPD markers (13 and 22% respectively). However, AFLP primers amplified on average six times more products than RAPD markers. The average numbers of polymorphic products per primer were 1.6 and 6.5 for RAPD and AFLP primers, respectively, i.e., AFLP primers were four times more efficient than RAPD primers in their ability to detect polymorphism in pepper. While four blocky type cultivars were indistinguishable by RAPD, two AFLP primer pairs were sufficient to distinguish the four cultivars from each other.
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  • 85
    ISSN: 1573-5060
    Keywords: embryo sac ; RAPD ; interspecific hybrid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract In cassava, apomixis fixes heterosis and avoids transmission of systemic pathogens which complicate vegetative propagation of the crop. A combination of evidence from maternal inheritance of RAPD markers and the structure of the embryonic sac in large progeny sets of two distinct genotypes have further confirmed the occurrence of apomixis in cassava. We could advance further on earlier reports of the detection of apomixis in four ways: (1) we could arrive at an estimate of the rate of facultative apomixis in the range of 2%; (2) we detected the occurrence of apomixis in a second genotype, derived from a different interspecific cross; (3) apomictic behavior was demonstrated in an F1 individual and (4) parallel embryonic evidence was generated that corroborate the potential occurrence of apomixis by apospory. The fact that apomixis was detected in an F1 interspecific hybrid hints to the possibility of directly transferring genes for apomixis from a wild relative to cultivated cassava.
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  • 86
    ISSN: 1573-5060
    Keywords: bottleneck ; isozyme ; landrace ; lentil ; RAPD ; South Asia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Lentil landraces from South Asia exhibit a low diversity and discordance with landraces from other countries according to a combination of qualitative and quantitative agromorphological characters. They exhibit specific phenological adaptation to the South Asian environment which precludes the direct use of alien germplasm in breeding programs in South Asia. An understanding of the genetic relationships and diversity of South Asian lentil landraces, in relation to landraces from other countries, is important in attempting to widen the genetic base of germplasm in the region. The objectives of this study were to investigate the genetic relationships between lentil landraces from 3 South Asian countries (India, Nepal and Pakistan) and those from 13 other countries and to determine their relative genetic diversities, using both isozyme electrophoresis and random amplified polymorphic DNA (RAPD) analysis. Polymorphisms were observed for 7 isozyme loci (16 alleles) and 22 RAPD loci. According to Nei's genetic distance, germplasm from Afghanistan clustered with that from the South Asian countries. The germplasm from these countries was striking different to that from the other countries studied. Based on genetic distance estimates from RAPD analysis, the countries with the lowest diversity were Pakistan, Afghanistan and Nepal. These data support evidence at the morphological level of a genetic bottleneck in lentil landraces from South Asia. Genetic relationships between countries outside the South Asian group are discussed. Classification into macrosperma and microsperma types did not reflect overall country relationships.
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  • 87
    ISSN: 1573-5060
    Keywords: chloroplast DNA ; eggplant ; PCR ; RFLP ; Solanum ; taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract RFLP analysis of a PCR amplified 3.2-kbp region of cpDNA bounded by the conserved sequences in rbc L and ORF 106 was performed in eggplant (Solanum melongena), its related Solanum species, S. incanum, S. virginianum (= S. surattense), S. torvum, S. aethiopicum (= S. gilo), S. aethiopicum (= S. integrifolium), S. violaceum (= S. indicum), S. violaceum (= S. sanitwongsei) and S. mammosum and the reciprocal hybrids between S. aethiopicum (= S. integrifolium) and S. melongena 'Uttara'. The target region of cpDNA was amplified correctly by PCR. The amplified products were digested with each of 10 restriction enzymes (Alu I, Ase I, BamH I, Hinf I, Msp I, Rsa I, ScrF I, Sty I, Taq I and Xba I). Variations of restriction patterns among the species were recognized after digesting the amplified products with each of the seven restriction enzymes, Taq I, Alu I, Rsa I, Sty I, Ase I, Hinf I and Xba I. The restriction patterns divided the examined nine species into the following five clusters, 1) S. melongena and S. incanum, 2) S. virginianum (= S. surattense), 3) S. torvum, 4) S. aethiopicum (= S. gilo), S. aethiopicum (= S. integrifolium), S. violaceum (= S. indicum) and S. violaceum (= S. sanitwongsei) and 5) S. mammosum. The restriction pattern with Alu I in each of the reciprocal hybrids between S. melongena 'Uttara' and S. aethiopicum (= S. integrifolium) was identical with that of seed parent. The present study demonstrated the availability of the PCR-RFLP analysis of cpDNA for assessing taxonomic relationships and identifying cytoplasmic parentage of interspecific hybrids in eggplant and related Solanum species.
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  • 88
    ISSN: 1573-5060
    Keywords: dihaploid Solanum tuberosum ; Phytophthora infestans (Mont.) de Bary ; polygenically inherited resistances ; somatic hybrids ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Five dihaploid Solanum tuberosum genotypes, encompassing different levels of polygenically inherited resistance to potato late blight disease (Phytophthora infestans (Mont.) de Bary) in foliage and tubers, were used in four intraspecific protoplast fusion combinations. Vigorous growing putative hybrid calli were selected four weeks after electrofusion. Intact plants were regenerated from 6% of the selected calli. Verification of hybridity was accomplished by use of RAPD analysis which revealed that 53% of the regenerated plants were true somatic hybrids. The score of true somatic hybrids in the different fusion combinations ranged within 21% – 100%. The hybrid plants were analysed for resistance to foliage blight in a field trial and assessed for tuber blight resistance by use of a laboratory test. Resistance to late blight in foliage and tubers varied between the hybrids. Very high levels of resistance to both foliage and tuber blight were obtained in some hybrids. However, loss of resistance in some hybrids as compared to the parental plants were also observed. Possible reasons for the phenotypic disappearance of resistance to either foliage or tuber blight are suggested.
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  • 89
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    Euphytica 103 (1998), S. 287-292 
    ISSN: 1573-5060
    Keywords: Sorghum ; genome mapping ; rust ; RFLP ; QTL
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The location and effects of genomic regions for rust resistance in sorghum were determined. One hundred and sixty recombinant inbreds, which derived from a cross between QL39 and QL41, were used as a segregating population for genome mapping and rust resistance evaluation. Phenotypic data were collected in replicated field trials in two years. Interval mapping and non-parametric mapping identified four regions, each in a separate linkage group, associated with rust resistance. The region with the largest effect on rust resistance is on linkage group 10; it accounted for 40% of the total phenotypic variation.
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  • 90
    ISSN: 1573-5060
    Keywords: taro ; Colocasia esculenta ; RAPD ; DNA fingerprinting ; phylogenetic tree
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Forty-four taro (Colocasia esculenta), two tanier (Xanthosoma species) and one Colocasia gigantea accessions were evaluated for genetic diversity using random amplified polymorphic DNA (RAPD) primers. Seventy-three of 112 primers amplified PCR DNA products used to fingerprint the accessions. Thirty-two primers were considered highly informative because they amplified more than 5 bands or amplified one or more polymorphic bands that distinguished between accessions. RAPDs showed high genetic diversity in taro accessions from Indonesia, were capable in distinguishing between Hawaiian accessions, and could separate triploid from diploid accessions. UPGMA cluster analysis of genetic similarity estimates (Jaccard's coefficient), separated the accessions into 3 main groups with C. esculenta divided into 5 subgroups. These primers will be useful for future genetic analysis and provide taro breeders with a genetic basis for selection of parents for crop improvement. Polymorphic markers identified in the DNA fingerprinting study will be useful to screen a segregating population which is being generated in our laboratory aimed at developing a taro genetic linkage map.
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  • 91
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    Genetic resources and crop evolution 45 (1998), S. 511-523 
    ISSN: 1573-5109
    Keywords: Allium ; crop evolution ; GISH ; hybridogenic cultivars ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Three vegetative crops of sect. Cepa in genus Allium (Top onion, French grey shallot and viviparous triploid onion) of suspected hybridogenic origin were studied with genomic in situ hybridization (GISH) and random amplified polymorphic DNA (RAPD) markers. Related wild and cultivated species were included in the analysis in order to assess their contributions to the genomes of the investigated species. In A. × proliferum, the parental chromosomes derived from A. fistulosum and A. cepa were unequivocally identified by GISH, proving the hybrid status of this crop. The French grey shallot proved to belong to A. oschaninii according to the RAPD analysis and the GISH results, it is clearly separate from the normal shallots of A. cepa var. aggregatum. Thus the grey shallots are a new crop species and can be considered as an aggregatum form of the wild progenitor species A. oschaninii. The triploid viviparous onion comprises mostly A. cepa derived DNA in its genome. A non-cepa component could not be attributed to any of the sect. Cepa species included in this study and is most likely derived from one of the species of this section not yet identified by molecular means.
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  • 92
    ISSN: 1573-5109
    Keywords: chloroplast DNA ; hybridization ; introgression ; Pyrus ; RFLP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Polymorphism of chloroplast DNA of 106 accessions of Pyrus (mainly east Asian accessions) was examined. Four haplotypes were observed with the combination of three independent restriction site mutations digested by EcoO109 I, Sal I, and Xba I, respectively. In the occidental pear accessions only the most plesiomorphic type was observed, whereas all four types appeared in the oriental pear accessions. This suggests that the oriental species of Pyrus and occidental ones may have evolved independently. The distribution of four haplotypes in the east Asian pear was quite incongruent with the species or infrageneric classification using mainly morphological characters. Considering the high crossability and much occurrence of suspected interspecific hybrids in wild populations, this disaccord is inferred to be the results of the hybridization and introgression between species.
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  • 93
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    Genetic resources and crop evolution 45 (1998), S. 243-251 
    ISSN: 1573-5109
    Keywords: Phaseolus ; genetic variability ; amino acids ; isozymes ; proteins ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The genetic variation within and between Spanish landraces or varieties of Phaseolus vulgaris L. (common bean) and P. coccineus L. (runner bean) has been estimated by means of isozymes and random amplified polymorphic DNA (RAPD) analyses. Likewise, storage protein and amino acid content in dry seeds have been estimated. Fifteen landraces (60 accessions) of P. vulgaris and six of P. coccineus (six accessions) have been studied. Of the seven isozymatic systems analyzed only three systems and three loci showed variability in each species. Isozyme analyses revealed that genetic variability within and between landraces exist in both species. Even variability within accession was detected in some P. vulgaris landraces. Comparison of isozyme data indicated that Spanish landraces have a lower level of genetic variability than wild American materials and probably also lower than American landraces. RAPD analysis allowed for the uniquely distinguishing of all landraces. Genetic similarity among landraces, estimated by both isozymes and RAPDs, were not related with the seed morphological characters (color, size and shape) which define each variety or landrace. Variation in protein and amino acid content among landraces was also detected. The average protein content in common bean (20.48%) was similar to values previously reported in this species and higher than the average in the runner bean landraces (16.33%). In relation to the amino acid content methionine and cysteine showed the lowest values in all samples, although the content of these two amino acids varied widely among landraces.
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  • 94
    ISSN: 1573-5109
    Keywords: DNA fingerprints ; genetic diversity ; Ipomoea batatas ; polymerase chain reaction ; RAPD ; sweetpotato
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The island of New Guinea is considered a secondary center on diversity for sweetpotato, because of its range of isolated ecological niches and large number of cultivars found within a small area. Information of genetic diversity in Papua New Guinea (PNG) sweetpotato is essential for rationalizing the global sweetpotato germplasm collection. Using random amplified polymorphic DNA (RAPD), we compared the genetic variation and genetic diversity in 18 PNG cultivars versus 18 cultivars from South America. The analysis of molecular variance revealed large genetic diversity in both groups of cultivars. The within-group (among individuals) variation accounted for 90.6% of the total molecular variance. However, the difference between PNG and South American groups is statistically significant, although it explained only 9.4% of the total molecular variance. The PNG cultivars are also less divergent than their South American ancestors as the mean genetic distance in PNG group is significantly smaller than that of South American group. The lower level of genetic diversity in PNG cultivars was also reflected by multidimensional scaling. This study shows that PNG cultivars, after many years of isolated evolution in an unique agro-ecological environment are substantially divergent from their ancestors in South America. The genetic diversity level in PNG cultivars is significantly lower than that in South American cultivars. It thus provides a baseline for continuing studies of genetic diversity in different sweetpotato gene pools.
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  • 95
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    Genetic resources and crop evolution 45 (1998), S. 337-342 
    ISSN: 1573-5109
    Keywords: burr medic ; genetic variability ; Medicago ; outcrossing rate ; RAPD ; selfing
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Although burr medic (Medicago polymorpha L.) is commonly considered a self-pollinating species, intrapopulational variation for morphological, biochemical and molecular markers is relatively high. To investigate whether part of this variation is the result of outcrossing, we designed RAPD analysis experiments to reveal both inter- and intra-accession crossing. No cases of inter-accession hybrids were documented, but an intra-accessional crossing rate of 0 to 4% was estimated for one of the four accessions studied. Therefore, a rare outcrossing event between local individuals and migrating genotypes may contribute to high genetic variability observed in natural M. polymorpha. A better understanding of the factors which influence outcrossing in M. polymorpha is pertinent both for medic breeding programmes and for assessing risks associated with releasing transgenic herbicide-resistant crops.
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  • 96
    ISSN: 1573-5109
    Keywords: AMOVA ; Lens ; population genetic structure ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract An understanding of the genetic structure of populations is vital for the formation of optimum collection, conservation and utilization strategies for plant genetic resources. This is of particular importance in the case of in-situ conservation, a strategy gaining in popularity. The population genetic structures of five wild lentil taxa, Lens culinaris subsp. orientalis, L. odemensis, L. ervoides, L. nigricans and L. lamottei were investigated using isozyme electrophoresis and random amplified polymorphic DNA (RAPD). Approximately 20 plants from each of 5 populations per taxon were screened for variation at 11 isozyme loci and using three RAPD primers. Levels of variation were generally low, although considerable variation existed in the levels of diversity found within populations of L. culinaris subsp. orientalis and L. lamottei. Comparison of the results obtained in this study with the results obtained in a previous study indicate that this is a trend occurring across all species. It implies that levels of diversity within populations must be measured and considered prior to targeting of specific populations for in-situ conservation. Analysis of molecular variance of both isozyme and RAPD data revealed that between 78% and 99% of the variation was attributable to between-population differences. Isozyme results from L. lamottei populations were, however, contradictory. Possible explanations for this difference are discussed.
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  • 97
    ISSN: 1573-5117
    Keywords: RAPD ; Littorina saxatilis ; L. neglecta ; phylogeny
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A genetic analysis of ‘morphotypes’ of Littorina saxatilis from two locations on the north-east coast of England (Filey and Ravenscar), using randomly amplified DNA polymorphisms (RAPD) generated with a single primer, revealed quite different patterns of variation. Thin shelled, wide-apertured (H-form) animals from Ravenscar tended to cluster separately from thick shelled (M) forms, indicating genetic differentiation of these morphs. Animals of similar morphology (H and M) from Filey (about 30 km distant) did not display such an obvious pattern, and although there was still evidence of differentiation from discriminant analysis of RAPD data, levels of correct classification were reduced at Filey. This suggests that the utility of a single RAPD primer for separation of such forms varies over a relatively small distance. L. arcana from Ravenscar, included as an outgroup, were generally well differentiated from L. saxatilis and were noted to exhibit less variation, a phenomenon that has been noted previously in some allozyme and RAPD analyses. A similar RAPD analysis undertaken on small, barnacle dwelling, brooding forms from Peak Steel, Ravenscar revealed that animals appeared to have as great a tendency to cluster together on a microgeographic scale (by collection patch) as by ‘species’ ( L. neglecta or L. saxatilis b) although predominance of certain species in individual patches largely explains this. Discriminant analysis of RAPD presence/absence data did correctly place over 90% of barnacle dwelling animals to their respective species, and we consider this as evidence of separate gene pools. RAPD is taken to be a useful tool for screening genetic variation in this complex of animals on a local scale when either a pre-selected informative primer is utilised or a battery of primers is used, but its efficacy may be reduced when a single primer is employed for screening animals from different shores.
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  • 98
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    Hydrobiologia 378 (1998), S. 33-42 
    ISSN: 1573-5117
    Keywords: protocols ; RAPD ; Littorina saxatilis ; L. obtusata ; L. fabalis ; L. littorea ; extracting DNA ; amplifying DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Randomly amplified polymorphic DNA (RAPD) is a fast and useful method of genome marking that is useful for studies of, for example, parentage, mating patterns, taxonomy of sibling species and intra-specific population genetic structures. Here we compare three different procedures for extracting high molecular weight genomic DNA; phenol-chloroform, hexadecyltrimethyl ammonium-bromide (CTAB) and Chelex 100. Double phenol-chloroform and CTAB extractions both generated high amounts of high quality DNA while Chelex 100 failed to do so. We also compared PCR-amplification with different concentrations of template DNA and found that 1–2 ng per 25 μl of amplification cocktail gave the best results. Amplifying DNA prepared by the three extraction methods revealed that DNA extracted with double phenol-chloroform gave the clearest bands. The double phenol-chloroform extraction seems thus the most suitable extraction method for RAPD in Littorina, however Chelex may be the only method useful for extracting DNA from very small individuals, for example, pre-hatching stages.
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  • 99
    ISSN: 1573-6857
    Keywords: genus Pellia ; liverworts ; RAPD ; taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The random amplified polymorphic DNA (RAPD) technique was used to study the genomic relationships of three liverworts from the genus Pellia: P. epiphylla, P. borealis and P. neesiana. Altogether 150 characters (150 DNA fragments obtained using PCR) were scored. These characters were used to create a matrix of pairwise distances between all the pairs of taxa. Both distance (UPGMA, Fitch–Margoliash and Neighbor–Joining) and binary character‐state (Wagner and Camin‐Sokal parsimony and compatibility) methods were applied for trees' construction. Our results strongly support distinction of the recently discovered sibling species of P. epiphylla – species N and P. epiphylla – species S, which have an allopatric distribution in Poland (N – North, S – South Poland). Moreover, our data also supports the hypothesis of a hybrid origin (alloploid) of the polyploid P. borealis from P.epiphylla−N×P.epiphylla−S. P. neesiana was excluded as a donor of either of the genomes of P. borealis.
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  • 100
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    World journal of microbiology and biotechnology 14 (1998), S. 415-420 
    ISSN: 1573-0972
    Keywords: Arbitrary primers ; Brucella abortus ; Brucella melitensis ; polymerase chain reaction ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Randomly amplified polymorphic DNA (RAPD) profiles of Brucella and non-Brucella DNA were established after polymerase chain reaction (PCR) amplification. Five arbitrary oligonucleotide primers were screened to generate Brucella-specific DNA fingerprints. The arbitrary primer OPB-01 (5′-GTTTCGCTCC-3′) produced DNA bands specific to Brucella. Amplification conditions must be optimized for reproductibility. Accordingly, we optimized and established the conditions, which included Mg2+, enzyme (DNA polymerase), primer, template and deoxyribonucleoside triphosphate (dNTP) concentrations as well as the optimum number of thermal cycles to produce OPB-01 directed Brucella DNA fingerprints. The optimized RAPD method can produce a 1.3 kb DNA fragment specific to Brucella. This DNA fragment was common to eight biovars of B. abortus and one biovar of B. melitensis. The fragment was not detected in genetically related species such as Ochrobactrum anthropi and other non-Brucella organisms associated with farm animals. We anticipate the use of this fragment as a possible probe for the detection of Brucella organisms.
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