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  • 1
    ISSN: 1432-2242
    Keywords: Key words Map ; Onion ; Allium ; Duplication
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The bulb onion, Allium cepa L., is a diploid (2n=2x=16) plant with a huge nuclear genome. Previous genetic and cytogenetic analyses have not supported a polyploid origin for onion. We developed a low-density genetic map of morphological markers, randomly amplified polymorphic DNAs (RAPD), and restriction fragment length polymorphisms (RFLP) as a tool for onion improvement and to study the genome organization of onion. A mapping population of 58 F3 families was produced from a single F1 plant from the cross of two partially inbred lines (Brigham Yellow Globe 15-23 and Alisa Craig 43). Segregations were established for restoration of male fertility in sterile cytoplasm, complementary light-red bulb color, 14 RAPDs, 110 RFLPs revealed by 90 anonymous cDNA clones, and 2 RFLPs revealed by a cDNA clone of alliinase, the enzyme responsible for the characteristic Allium flavors. Duplicated RFLP loci were detected by 21% of the clones, of which 53% were unlinked (〉30 cM), 5% loosely linked (10–30 cM), and 42% tightly linked (〈10 cM). This duplication frequency is less than that reported for paleopolyploids but higher than for diploid species. We observed 40% dominant RFLPs, the highest yet reported among plants. Among duplicated RFLP loci, 19% segregated as two loci each with two codominant alleles, 52% segregated as one locus with codominant alleles and one locus with only a dominant fragment, and 29% segregated as two loci with only dominant fragments. We sequenced cDNAs detecting duplicated RFLPs; 63% showed homology to known gene families (e.g., chlorophyll binding proteins, ubiquitin, or RuBISCO), and 37% were unique clones showing significant homology to known genes of low-copy number or no homology to database sequences. Duplicated RFLPs showing linkage could be due to retroviral-like sequences in adjacent coding regions or intrachromosomal, as opposed to whole genome, duplications. Previous cytological analyses and this genetic map support intrachromosomal duplication as a mechanism contributing to the huge onion genome.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Theoretical and applied genetics 97 (1998), S. 960-967 
    ISSN: 1432-2242
    Keywords: Key words AFLP ; Bulked segregant analysis ; Daucus carota ; Inverse PCR ; Marker conversion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Recent advances have expanded the potential usefulness of molecular techniques for plant genetic research. AFLP is a powerful technique, allowing rapid and reliable analysis of multiple, potentially polymorphic sites in a single experiment. Because AFLP technology requires no a priori knowledge of genome structure or preparation of molecular probes, it is immediately useful for a wide variety of plant species. However, because AFLP markers are dominant, costly, and technologically demanding, the technique has limited application for large-scale, locus-specific uses. In carrot, the Y 2 locus controls carotene accumulation in the root xylem core. Although carrot is an important source of dietary carotene, little is known about the regulation and biosynthesis of carotenes in carrot. We identified six AFLP fragments linked to the Y 2 locus through a combination of F2 mapping and bulked segregant analysis. We have developed a procedure for generating simple, codominant, PCR-based markers from dominant AFLP fragments using a Y 2 -linked AFLP fragment as a model. Our converted marker requires only a simple PCR followed by standard agarose gel electrophoresis. It is rapid, simple, reliable, comparatively inexpensive, codominant, and non-radioactive. Conversion of AFLP fragments to forms better adapted to large-scale, locus-specific applications greatly expands the usefulness of this molecular technique.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-2242
    Keywords: Key words Solanum bulbocastanum ; Late blight ; Phytophthora infestans ; Somatic hybrid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Somatic hybrids between potato and Solanum bulbocastanum, a wild diploid (2n=2x=24) Mexican species, are highly resistant to late blight, caused by Phytophthora infestans. Both randomly amplified polymorphic DNA (RAPD) and restriction fragment length polymorphism (RFLP) markers that are closely linked to the resistance have been noted by analysis of three different backcross-2 populations derived from two different somatic hybrids. With reference to previously published potato and tomato maps, resistance appears to be on the long arm of chromosome 8 and is flanked by RFLP markers CP53 and CT64. In a population of BC2 plants derived from a cross between the BC1 line J10lK6 [(S. tuberosum PI 203900+S. bulbocastanum PI 243510) ×Katahdin)]×Atlantic, late blight resistance cosegregated with RFLP marker CT88 and RAPD marker OPG02–625.
    Type of Medium: Electronic Resource
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