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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Theoretical and applied genetics 91 (1995), S. 557-562 
    ISSN: 1432-2242
    Keywords: Solanum tuberosum ; Phytophthora infestans ; Transposable element ; Transformation competence ; Linkage analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In the course of improving diploid potato genotypes for transformation ability, selection for specific components affecting regeneration and transformation was carried out. From a segregating population between two good regenerating clones a selection was made to yield an optimal well-transforming and fertile genotype J92-6400-A16. This plant yielded predominantly diploid transformants and was heterozygous for the gene R1, conferring resistance to Phytophthora infestans. The speed of, and competence for, regeneration and transformation on both sides of the stem explant were improved. A competence factor for tranformation was found to be linked with the R1 locus and a molecular marker on chromosome 5. The male fertility of transformants was frequently decreased to a great extent, whereas female fertility was not so markedly affected.
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  • 2
    ISSN: 1432-2242
    Keywords: Key words Ac/Ds transposon tagging ; Somatic selection ; Hygromycin ; Phytophthora infestans ; R1 resistance gene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Potato is an autotetraploid crop plant that is not very amenable to the deployment of transposon tagging for gene cloning and gene identification. After diploidisation it is possible to get potato genotypes that grow well, but they are self-incompatible. This prevents the production of selfed progeny that are normally used in gene tagging approaches to select for parental lines with the target gene to be tagged in a homozygous stage. We describe here an alternative selection method for directed transposon tagging for a gene of interest in a heterozygous background. Diploid potato plants with a Ds transposon linked to the desired gene of interest (the Phytophthora infestans R1 resistance locus) in a heterozygous stage were used for the development of this directed transposon tagging strategy. After crossing to a diploid Ac transposon-containing genotype, 22 ’interesting’ seedlings (R1Ds/r–; Ac/–) were selected that showed active Ds transposition as displayed by DNA blot hybridisation, empty donor site PCR and sequencing. Protoplast isolation and the use of the hygromycin gene as a cell-specific selection marker of Ds excision enabled the direct selection of Ds excision sectors in these highly chimaeric seedlings. This somatic selection of Ds transpositions and the regeneration through protoplasts resulted in the development of a large population of almost 2000 hygromycin-resistant plants. Southern blot analysis confirmed the insertion of Ds at independent positions in the genome. Every selected plant displayed independent Ds excisions and re-insertions due to the expression of the Ac transposase throughout development. This population, which is developed from seedlings with the desired R1 gene in a heterozygous stage, is directly useful for searching for transposon-tagged R1 mutants. In general, this approach for selecting for somatic transpositions is particularly suitable for the molecular isolation of genes in a heterozygous crop like potato.
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  • 3
    ISSN: 1617-4623
    Keywords: Potato ; Solanum tuberosum ; Phytophthora infestans ; RFLP ; R genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Phytophthora infestans (Mont.) de Bary is the most important fungal pathogen of the potato (Solanum tuberosum). The introduction of major genes for resistance from the wild species S. demissum into potato cultivars is the earliest example of breeding for resistance using wild germplasm in this crop. Eleven resistance alleles (R genes) are known, differing in the recognition of corresponding avirulence alleles of the fungus. The number of R loci, their positions on the genetic map and the allelic relationships between different R variants are not known, except that the R1 locus has been mapped to potato chromosome V The objective of this work was the further genetic analysis of different R alleles in potato. Tetraploid potato cultivars carrying R alleles were reduced to the diploid level by inducing haploid parthenogenetic development of 2n female gametes. Of the 157 isolated primary dihaploids, 7 set seeds and carried the resistance alleles R1, R3 and R10 either individually or in combinations. Independent segregation of the dominant R1 and R3 alleles was demonstrated in two F1 populations of crosses among a dihaploid clone carrying R1 plus R3 and susceptible pollinators. Distorted segregation in favour of susceptibility was found for the R3 allele in 15 of 18 F1 populations analysed, whereas the RI allele segregated with a 1:1 ratio as expected in five F1 populations. The mode of inheritance of the R10 allele could not be deduced as only very few F1 hybrids bearing R10 were obtained. Linkage analysis in two F1 populations between R1, R3 and RFLP markers of known position on the potato RFLP maps confirmed the position of the R1 locus on chromosome V and localized the second locus, R3, to a distal position on chromdsome XI.
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