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  • 11
    ISSN: 1432-2242
    Keywords: Potato cyst-nematode ; Quantitative trait loci RFLP ; Distorted segregation ; Solanum spegazzinii
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A high level of resistance toGlobodera pallida pathotypes Pa2 and Pa3 exists inSolanum spegazzinii, a wild relative of potato (S. tuberosum ssp.tuberosum). Here we report the mapping of loci involved in quantitatively-inherited nematode resistance with the use of RFLPs. One major locus,Gpa, was mapped on chromosome 5 and two minor loci on chromosomes 4 and 7 ofS. spegazzinii. Additionally, the contribution of the susceptible parent to nematode resistance was determined. TheGpa locus was solely responsible for the high resistance level found in the segregating population. However, the RFLP marker closely linked to this resistance locus showed a distorted segregation, with a shortage of plants having the resistance linked allele. Our results indicate that a prediction of the genetic constitution of a quantitative trait based solely on phenotypic observations can lead to erroneous conclusions.
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  • 12
    ISSN: 1432-2242
    Keywords: Molecular markers Beet cyst nematode resistance ; Hs1 Heterodera schachtii Schm ; Satellite DNA Monosomic fragment additions Beta ; Pulsed-field gel electrophoresis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract New members of a satellite DNA family (Sat 121), specific for wild beets of the section Procumbentes of the genus Beta, were isolated. Sequence analysis showed that the members of Sat-121 fall into two distinct classes. The organization of Sat-121 in the vicinity of a beet cyst nematode (Heterodera schachtii Schm.) resistance locus (Hs1) in B. patellaris and B. procumbens was investigated by pulsed-field gel electrophoresis (PFGE) using DNA from a series of resistant monosomic fragment additions, each containing an extra chromosome fragment of B. patellaris chromosome-1 (pat-1) in B. vulgaris. In this way several clusters of Sat-121 flanking the Hs1 pat-1 locus were identified. In nematode resistant diploid introgressions (2n=18), which contain small segments of B. procumbens chromosome-1 (pro-1) in B. vulgaris, only two major Sat-121 clusters were detected near the Hs1 pro-1 locus.
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  • 13
    Electronic Resource
    Electronic Resource
    Springer
    Theoretical and applied genetics 92 (1996), S. 463-470 
    ISSN: 1432-2242
    Keywords: Key words  RFLPs ; Solanum spegazzinii ; Globodera sp. ; Interlocus interaction ; Intralocus interaction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract   A backcross population, derived from the cross (S. tuberosum×S. spegazzinii)×S. tuberosum was used to map QTLs involved in nematode resistance, tuber yield and root development. Complete linkage maps were available for the interspecific hybrid parent as well as the S. tuberosum parent, and interval mapping for all traits was performed for both. Additionally, the intra- and inter-locus interactions of the QTLs were examined. The Gro1.2 locus, involved in resistance to G. rostochiensis pathotype Ro1, that was previously mapped in the S. tuberosum×S. spegazzinii F1 population, was located more precisely on chromosome 10. A new resistance locus, Gro1.4, also conferring resistance to G. rostochiensis pathotype Ro1, was found on chromosome 3. Different alleles of this locus originating from both parents contributed to the resistant phenotype, indicating multiallelism at this locus. No interlocus interactions were observed between these two resistance loci. For resistance to G. pallida no QTLs were detected. One minor QTL involved in tuber yield was located on chromosome 4. Two QTLs involved in root development and having large effects were mapped on chromosomes 2 and 6 and an epistatic interaction was found between these two loci.
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  • 14
    ISSN: 1432-2242
    Keywords: Randomly amplified polymorphic DNA (RAPD) ; Deletion mapping ; Sequence Tagged Site (STS) ; Monosomic fragment additions ; Beet cyst nematode resistance ; Heterodera schachtii Schm ; Beta patellaris
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A beet cyst nematode (BCN)-resistant telosomic addition of B. patellaris chromosome 1 in B. vulgaris was used to isolate 6 RAPD markers linked to the BCN resistance locus Hs1 pat-1. Southern analysis showed that the analyzed RAPD products contain either low-, middle or high-repetitive DNA. The relative positions of the random amplified polymorphic DNA (RAPD) markers and of the restriction fragment length polymorphism (RFLP) loci corresponding to the low-repetitive RAPD products were determined by deletion mapping using a panel of seven nematode-resistant B. patellaris chromosome-1 fragment additions. One RAPD marker, OPB11800, was found to be present in two copies on the long arm telosome of B. patellaris chromosome 1. These copies are closely linked to the BCN resistance gene and flank the gene on both sides. On the basis of the nucleotide sequence of OPB11800, sequence-tagged site (STS) primers were developed that amplify specific fragments derived from the two OPB11800 loci. These STS markers can be used in the map-based cloning of the BCN gene, as they define start and finishing points of a chromosomal walk towards the Hs1 pat-1 locus. Two copies of the middle-repetitive OPX21100 marker were mapped in the same interval of the deletion mapping panel as the resistance gene locus and thereby belong to the nearest markers as yet found for the BCN gene in B. patellaris.
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  • 15
    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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  • 16
    Electronic Resource
    Electronic Resource
    Springer
    Theoretical and applied genetics 92 (1996), S. 463-470 
    ISSN: 1432-2242
    Keywords: RFLPs ; Solanum spegazzinii ; Globodera sp ; Interlocus interaction ; Intralocus interaction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A backcross population, derived from the cross (S. tuberosumxS. spegazzinii)xS. tuberosum was used to map QTLs involved in nematode resistance, tuber yield and root development. Complete linkage maps were available for the interspecific hybrid parent as well as the S. tuberosum parent, and interval mapping for all traits was performed for both. Additionally, the intra- and inter-locus interactions of the QTLs were examined. The Gro1.2 locus, involved in resistance to G. rostochiensis pathotype Ro1, that was previously mapped in the S. tuberosumxS. spegazzinii F1 population, was located more precisely on chromosome 10. A new resistance locus, Gro1.4, also conferring resistance to G. rostochiensis pathotype Ro1, was found on chromosome 3. Different alleles of this locus originating from both parents contributed to the resistant phenotype, indicating multiallelism at this locus. No interlocus interactions were observed between these two resistance loci. For resistance to G. pallida no QTLs were detected. One minor QTL involved in tuber yield was located on chromosome 4. Two QTLs involved in root development and having large effects were mapped on chromosomes 2 and 6 and an epistatic interaction was found between these two loci.
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  • 17
    ISSN: 1432-2242
    Keywords: Potato ; Agrobacterium transformation ; Plant regeneration ; Monohaploid, diploid ; Genetic markers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Agrobacterium transformation of stem internodes of four monohaploid (839-79, 849-7, 851-23, 855-1) and two diploid (M9 and HH260) potato genotypes using hairy root-inducing single (LBA 1020, LBA 9365, LBA 9402) and binary (LBA 1060KG) vectors is reported. Various media and successive culture steps were tested for plant regeneration from different transformed root clones. The fate of introduced genetic markers in root clones and regenerated plants (hairy root phenotype, hormone autotrophy, opine production, kanamycin resistance, β-glucuronidase activity), the ploidy stability and protoplast yield were analysed. The transformation efficiency of stem internodes (hairy root production) and the regeneration capacity of the transformed root clones greatly differed within and between the various potato genotypes. The regenerated plants obtained after transformation with both types of vectors often showed the absence of one or more genetic markers. However, transformation with the binary Agrobacterium vector generally resulted in the stable presence of the opines in all transformed root clones and most regenerated plants. In HH260, transformation efficiency, plant regeneration of transformed root clones, protoplast yield and ploidy stability were the highest as compared to the other genotypes. The application of these transformed plants as marker lines in gene mapping and gene expression studies is indicated.
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  • 18
    ISSN: 1432-2242
    Keywords: Nematode resistance Pulsed field electrophoresis ; Satellite DNA ; Sugar beet Yeast artificial chromosome library
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A YAC library was constructed from the Beta vulgaris fragment addition AN5-203b. This monosomic fragment addition harbors an approximate 12-Mbp fragment of B.patellaris chromosome 1 accomodating the Hs1 pat-1 conferring resistance to the beet cyst nematode (Heterodera schachtii). The YAC library consists of 20,000 YAC clones having an average size of 140 kb. Screening with organelle-specific probes showed that 12% of the clones contain chloroplast DNA while only 0.2% of the clones hybridizes with a mitochondrial specific probe. On the basis of a sugar beet haploid genome size of 750 Mbp this library represents 3.3 haploid genome equivalents. The addition fragment present in AN5-203b harbors a major satellite DNA cluster that is tightly linked to the Hs1 pat-1 locus. The cluster is located on a single 250-kb EcoRI restriction fragment and consists of an estimated 700–800 copies of a 159-bp core sequence, most of which are arranged in tandem. Using this core sequence as a probe, we were able to isolate 1 YAC clone from the library that contains the entire 250-kb satellite DNA cluster.
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  • 19
    ISSN: 1432-2242
    Keywords: Morphological markers ; Isozymes ; Non-inbred species ; Combined map ; JoinMap
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A genetic map of potato (Solanum tuberosum L.) integrating molecular markers with morphological and isozyme markers was constructed using a backcross population of 67 diploid potato plants. A general method for map construction is described that differs from previous methods employed in potato and other outbreeding plants. First, separate maps for the female and male parents were constructed. The female map contained 132 markers, whereas the male map contained 138 markers. Second, on the basis of the markers in common the two integrated parental maps were combined into one with the computer programme JoinMap. This combined map consisted of 175 molecular markers, 10 morphological markers and 8 isozyme markers. Ninety-two of the molecular markers were derived from DNA sequences flanking either T-DNA inserts in potato or reintegrated maize transposable elements originating from these T-DNA constructs. Clusters of distorted segregation were found on chromosomes 1,2,8 and 11 for the male parent and chromosome 5 for both parents. The total length of the combined map is 1120 cM.
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  • 20
    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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