Identification and molecular analysis of transgenic potato chromosomes transferred to tomato through microprotoplast fusion

E. Rutgers, K.S. Ramulu, P. Dijkhuis, J. Blaas, F.A. Krens, H.A. Verhoeven

    Research output: Contribution to journalArticleAcademicpeer-review

    13 Citations (Scopus)

    Abstract

    Results are reported on the integration sites and copy number of alien marker genes neomycin phosphotransferase II (nptII) and β-glucuronidase (uidA), introduced into diploid potato Solanum tuberosum through transformation by Agrobacterium tumefaciens. Also, the transgenic potato chromosomes 3 and 5 harbouring the nptII and uidA genes, which were transferred to tomato (wild species Lycopersicon peruvianum) by microprotoplast fusion, as revealed by genomic in situ hybridization (GISH), were identified by RFLP analysis using chromosome-specific markers. The data revealed three integration sites in the donor potato genome, each containing the uidA gene, and two also harbouring the nptII gene. Analysis of monosomic-addition hybrid plants obtained after microprotoplast fusion showed that each of these three integration sites is located on a different potato chromosome. The microprotoplast hybrid plants contained only the chromosomes that carried the selectable gene nptII. The data on sexual transmission of the donor potato chromosome carrying the uidA and nptII genes were obtained by analyzing the first backcross progeny (BC1) derived from crossing a monosomic-addition hybrid plant to tomato (L. peruvianum). The glucuronidase (GUS) assay and PCR analysis using primers for the uidA gent indicated the presence of the potato chromosome in GUS-positive and its absence in GUS-negative BC1 plants. RFLP analysis confirmed sexual transmission of the potato chromosome carrying the nptII and uidA genes to the BC1 plants. A few BC1 plants contained the nptII and uidA genes in the absence of the potato additional chromosome, indicating that the marker genes were integrated into the tomato genome. The potential applications of the transfer of alien chromosomes and genes by microprotoplast fusion technique are discussed.
    Original languageEnglish
    Pages (from-to)1053-1059
    JournalTheoretical and Applied Genetics
    Volume94
    Issue number8
    DOIs
    Publication statusPublished - 1997

    Fingerprint

    Kanamycin Kinase
    kanamycin kinase
    Lycopersicon esculentum
    Solanum tuberosum
    Chromosomes
    genetically modified organisms
    tomatoes
    potatoes
    chromosomes
    Glucuronidase
    Genes
    genes
    Solanum peruvianum
    monosomics
    Genetic Markers
    Restriction Fragment Length Polymorphisms
    restriction fragment length polymorphism
    Tissue Donors
    Genome
    genetic markers

    Cite this

    @article{cf00375ae708466d9aa0366172b75717,
    title = "Identification and molecular analysis of transgenic potato chromosomes transferred to tomato through microprotoplast fusion",
    abstract = "Results are reported on the integration sites and copy number of alien marker genes neomycin phosphotransferase II (nptII) and β-glucuronidase (uidA), introduced into diploid potato Solanum tuberosum through transformation by Agrobacterium tumefaciens. Also, the transgenic potato chromosomes 3 and 5 harbouring the nptII and uidA genes, which were transferred to tomato (wild species Lycopersicon peruvianum) by microprotoplast fusion, as revealed by genomic in situ hybridization (GISH), were identified by RFLP analysis using chromosome-specific markers. The data revealed three integration sites in the donor potato genome, each containing the uidA gene, and two also harbouring the nptII gene. Analysis of monosomic-addition hybrid plants obtained after microprotoplast fusion showed that each of these three integration sites is located on a different potato chromosome. The microprotoplast hybrid plants contained only the chromosomes that carried the selectable gene nptII. The data on sexual transmission of the donor potato chromosome carrying the uidA and nptII genes were obtained by analyzing the first backcross progeny (BC1) derived from crossing a monosomic-addition hybrid plant to tomato (L. peruvianum). The glucuronidase (GUS) assay and PCR analysis using primers for the uidA gent indicated the presence of the potato chromosome in GUS-positive and its absence in GUS-negative BC1 plants. RFLP analysis confirmed sexual transmission of the potato chromosome carrying the nptII and uidA genes to the BC1 plants. A few BC1 plants contained the nptII and uidA genes in the absence of the potato additional chromosome, indicating that the marker genes were integrated into the tomato genome. The potential applications of the transfer of alien chromosomes and genes by microprotoplast fusion technique are discussed.",
    author = "E. Rutgers and K.S. Ramulu and P. Dijkhuis and J. Blaas and F.A. Krens and H.A. Verhoeven",
    year = "1997",
    doi = "10.1007/s001220050514",
    language = "English",
    volume = "94",
    pages = "1053--1059",
    journal = "Theoretical and Applied Genetics",
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    Identification and molecular analysis of transgenic potato chromosomes transferred to tomato through microprotoplast fusion. / Rutgers, E.; Ramulu, K.S.; Dijkhuis, P.; Blaas, J.; Krens, F.A.; Verhoeven, H.A.

    In: Theoretical and Applied Genetics, Vol. 94, No. 8, 1997, p. 1053-1059.

    Research output: Contribution to journalArticleAcademicpeer-review

    TY - JOUR

    T1 - Identification and molecular analysis of transgenic potato chromosomes transferred to tomato through microprotoplast fusion

    AU - Rutgers, E.

    AU - Ramulu, K.S.

    AU - Dijkhuis, P.

    AU - Blaas, J.

    AU - Krens, F.A.

    AU - Verhoeven, H.A.

    PY - 1997

    Y1 - 1997

    N2 - Results are reported on the integration sites and copy number of alien marker genes neomycin phosphotransferase II (nptII) and β-glucuronidase (uidA), introduced into diploid potato Solanum tuberosum through transformation by Agrobacterium tumefaciens. Also, the transgenic potato chromosomes 3 and 5 harbouring the nptII and uidA genes, which were transferred to tomato (wild species Lycopersicon peruvianum) by microprotoplast fusion, as revealed by genomic in situ hybridization (GISH), were identified by RFLP analysis using chromosome-specific markers. The data revealed three integration sites in the donor potato genome, each containing the uidA gene, and two also harbouring the nptII gene. Analysis of monosomic-addition hybrid plants obtained after microprotoplast fusion showed that each of these three integration sites is located on a different potato chromosome. The microprotoplast hybrid plants contained only the chromosomes that carried the selectable gene nptII. The data on sexual transmission of the donor potato chromosome carrying the uidA and nptII genes were obtained by analyzing the first backcross progeny (BC1) derived from crossing a monosomic-addition hybrid plant to tomato (L. peruvianum). The glucuronidase (GUS) assay and PCR analysis using primers for the uidA gent indicated the presence of the potato chromosome in GUS-positive and its absence in GUS-negative BC1 plants. RFLP analysis confirmed sexual transmission of the potato chromosome carrying the nptII and uidA genes to the BC1 plants. A few BC1 plants contained the nptII and uidA genes in the absence of the potato additional chromosome, indicating that the marker genes were integrated into the tomato genome. The potential applications of the transfer of alien chromosomes and genes by microprotoplast fusion technique are discussed.

    AB - Results are reported on the integration sites and copy number of alien marker genes neomycin phosphotransferase II (nptII) and β-glucuronidase (uidA), introduced into diploid potato Solanum tuberosum through transformation by Agrobacterium tumefaciens. Also, the transgenic potato chromosomes 3 and 5 harbouring the nptII and uidA genes, which were transferred to tomato (wild species Lycopersicon peruvianum) by microprotoplast fusion, as revealed by genomic in situ hybridization (GISH), were identified by RFLP analysis using chromosome-specific markers. The data revealed three integration sites in the donor potato genome, each containing the uidA gene, and two also harbouring the nptII gene. Analysis of monosomic-addition hybrid plants obtained after microprotoplast fusion showed that each of these three integration sites is located on a different potato chromosome. The microprotoplast hybrid plants contained only the chromosomes that carried the selectable gene nptII. The data on sexual transmission of the donor potato chromosome carrying the uidA and nptII genes were obtained by analyzing the first backcross progeny (BC1) derived from crossing a monosomic-addition hybrid plant to tomato (L. peruvianum). The glucuronidase (GUS) assay and PCR analysis using primers for the uidA gent indicated the presence of the potato chromosome in GUS-positive and its absence in GUS-negative BC1 plants. RFLP analysis confirmed sexual transmission of the potato chromosome carrying the nptII and uidA genes to the BC1 plants. A few BC1 plants contained the nptII and uidA genes in the absence of the potato additional chromosome, indicating that the marker genes were integrated into the tomato genome. The potential applications of the transfer of alien chromosomes and genes by microprotoplast fusion technique are discussed.

    U2 - 10.1007/s001220050514

    DO - 10.1007/s001220050514

    M3 - Article

    VL - 94

    SP - 1053

    EP - 1059

    JO - Theoretical and Applied Genetics

    JF - Theoretical and Applied Genetics

    SN - 0040-5752

    IS - 8

    ER -