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  • 1
    Electronic Resource
    Electronic Resource
    An improved method for protoplast micro-injection suitable for transfer of entire plant chromosomes
    Amsterdam : Elsevier
    Plant Science 50 (1987), S. 161-169 
    Details
    ISSN: 0168-9452
    Keywords: Nicotiana plumbaginifolia ; chromosome transfer ; microinjection ; protoplasts
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology
    Type of Medium: Electronic Resource
    URL: http://linkinghub.elsevier.com/retrieve/pii/0168-9452(87)90153-1
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  • 2
    Electronic Resource
    Electronic Resource
    Microprotoplast isolation, enrichment and fusion for partial genome transfer in plants (1992)
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 85 (1992), S. 0 
    Details
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Somatic hybridization using protoplasts with whole genomes has often resulted in complex hybrids with many unwanted chromosomes or genes. Several researchers have attempted to reduce the number of undesired chromosomes through irradiation of the donor protoplasts, but so far without much success. Alternatively, micropro-toplasts containing one or a few chromosomes can be used for partial genome transfer, as has been demonstrated in human and other mammalian cell systems using microcells. Recently, we have optimized the ‘microprotoplast system’ for several donor cell lines (potato, Nicotiana, sugar beet) carrying various genetic markers, such as kanamycin resistance, β-glucuronidase, nitrate reductase deficiency, hormone autotrophy, opines, etc. Protocols were developed to obtain higher yields of micro-protoplasts as well as to enrich sub-diploid microprotoplasts containing one or a few chromosomes. These microprotoplasts were fused with whole mesophyll protoplasts of recipient lines using polyethylene glycol. Various requirements for lines used as donor and recipient partners in microprotoplast-protoplast fusions are described. The results are discussed in the context of partial genome transfer.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1399-3054.1992.tb04740.x
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  • 3
    Electronic Resource
    Electronic Resource
    Intergeneric transfer of a partial genome and direct production of monosomic addition plants by microprotoplast fusion (1996)
    Springer
    Theoretical and applied genetics 92 (1996), S. 316-325 
    Details
    ISSN: 1432-2242
    Keywords: Single ; specific chromosomes ; Transgenes ; Microprotoplast fusion ; Monosomic additions ; Genomic in situ hybridization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Results are reported on the transfer of single, specific chromosomes carrying kanamycin resistance (KanR) and β-glucuronidase (GUS) traits from a transformed donor line of potato (Solanum tuberosum) to a recipient line of the tomato species Lycopersicon peruvianum through microprotoplast fusion. Polyethylene glycol-induced mass fusion between donor potato microprotoplasts containing one or a few chromosomes and normal recipient diploid L. peruvianum protoplasts gave several KanR calli. A high frequency of plants regenerated from KanR calli expressed both KanR and GUS, and contained one or two copies of npt-II and a single copy of gus. Genomic in situ hybridization showed that several microprotoplast hybrid plants had one single potato donor chromosome carrying npt-II and gus genes and the complete chromosome complement of the recipient L. peruvianum (monosomic additions). Several monosomic-addition hybrid plants could be regenerated within the short time of 3 months and they were phenotypically normal, resembling the recipient line. These results suggest that the transfer of single chromosomes is tolerated better than is the transfer of the whole donor genome. The unique advantages of microprotoplast fusion are discussed: these include the direct production of monosomic addition lines for the transfer and introgression of economically important traits in sexually-incongruent species, the construction of chromosome-specific DNA libaries, high-resolution physical mapping and the identification of alien chromosome domains related to gene expression.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00223674
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  • 4
    Electronic Resource
    Electronic Resource
    Flow-cytometric characterization and sorting of plant chromosomes (1984)
    Springer
    Theoretical and applied genetics 67 (1984), S. 463-467 
    Details
    ISSN: 1432-2242
    Keywords: Chromosome isolation ; Flow cytometry ; Haplopappus gracilis (Nutt.) Gray ; Synchronization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Flow cytometric measurements of DNA frequency distribution were used to follow the synchronization process in suspension cells from Haplopappus gracilis (2n=4). Metaphase chromosomes were isolated from these synchronized cells and both the acro- and metacentric chromosomes were sorted by flow cytometry based on the different DNA contents. Possible applications of this procedure in fundamental genetics as well as practical plant breeding are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00263414
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  • 5
    Electronic Resource
    Electronic Resource
    Do S allele-specific peroxidase isoenzymes exist in self-incompatible Nicotiana alata? (1980)
    Springer
    Theoretical and applied genetics 57 (1980), S. 119-123 
    Details
    ISSN: 1432-2242
    Keywords: Self-incompatibility ; Nicotiana alata ; S Alleles-peroxidase isoenzymes-Styles ; Pollen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In order to test Pandey's hypothesis that peroxidase isoenzymes determine S-gene specificity in Nicotiana alata, peroxidase isoenzymes in styles and pollen from various plants of an inbred- and a cross progeny were compared by means of starch gel electrophoresis and electrofocusing. No relation between the S-genotype and the peroxidase isoenzyme patterns of pollen or of styles could be established. The differences between the isoenzyme patterns of different S-genotypes were ascribed to differences in the genetic background of various plants that had the same S-genotype.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00253883
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  • 6
    Electronic Resource
    Electronic Resource
    S-specific proteins in styles of self-incompatible Nicotiana alata (1981)
    Springer
    Theoretical and applied genetics 59 (1981), S. 185-190 
    Details
    ISSN: 1432-2242
    Keywords: Self incompatibility ; Nicotiana alata ; Sallele specific proteins ; Styles
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A comparison of the stigma protein patterns of individual plants of the inbred- and cross-progenies in Nicotiana alata by isoelectric focusing revealed the presence of S-specific proteins. The S allele-protein relationship was found for three different S alleles. The S-specific proteins occurred in both stigma and stylar parts of the pistil whereas they were absent in leaves. In clone OWL the concentration of S-specific proteins in the stigma increased gradually during floral development. The shift from compatibility to incompatibility was not accompanied by an abrupt increase in concentration of the S-proteins.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00264974
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  • 7
    Electronic Resource
    Electronic Resource
    Intergeneric transfer of a partial genome and direct production of monosomic addition plants by microprotoplast fusion (1996)
    Springer
    Theoretical and applied genetics 92 (1996), S. 316-325 
    Details
    ISSN: 1432-2242
    Keywords: Key words Single ; specific chromosomes  ; Transgenes  ;  Microprotoplast fusion  ;  Monosomic additions  ;  Genomic in situ hybridization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Results are reported on the transfer of single, specific chromosomes carrying kanamycin resistance and β-glucuronidase (GUS) traits from a transformed donor line of potato (Solanum tuberosum) to a recipient line of the tomato species Lycopersicon peruvianum through microprotoplast fusion. Polyethylene glycol-induced mass fusion between donor potato microprotoplasts containing one or a few chromosomes and normal recipient diploid L. peruvianum protoplasts gave several calli. A high frequency of plants regenerated from calli expressed both and GUS, and contained one or two copies of npt-II and a single copy of gus. Genomic in situ hybridization showed that several microprotoplast hybrid plants had one single potato donor chromosome carrying npt-II and gus genes and the complete chromosome complement of the recipient L. peruvianum (monosomic additions). Several monosomic-addition hybrid plants could be regenerated within the short time of 3 months and they were phenotypically normal, resembling the recipient line. These results suggest that the transfer of single chromosomes is tolerated better than is the transfer of the whole donor genome. The unique advantages of microprotoplast fusion are discussed: these include the direct production of monosomic addition lines for the transfer and introgression of economically important traits in sexually-incongruent species, the construction of chromosome-specific DNA libaries, high-resolution physical mapping and the identification of alien chromosome domains related to gene expression.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s001220050130
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  • 8
    Electronic Resource
    Electronic Resource
    Identification and molecular analysis of transgenic potato chromosomes transferred to tomato through microprotoplast fusion (1997)
    Springer
    Theoretical and applied genetics 94 (1997), S. 1053-1059 
    Details
    ISSN: 1432-2242
    Keywords: Key words Microprotoplast fusion ; Chromosome transfer ; Alien gene integration ; Chromosome identification ; Sexual transmission
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: 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 analysing 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 gene 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.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s001220050514
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  • 9
    Electronic Resource
    Electronic Resource
    Interspecific transfer of isolated plant mitochondria by microinjection (1995)
    Springer
    Plant cell reports 14 (1995), S. 781-785 
    Details
    ISSN: 1432-203X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Data on isolation, purification and transfer of mitochondria from a cytoplasmic male sterile line of the “Ogura” type of Raphanus sativus to a male fertile line of Brassica napus are reported. Microinjection has been used for the transfer of the donor mitochondria to the recipient protoplasts. The injected protoplasts were identified and followed individually throughout their development using a computerized microscope stage which greatly enhanced the number of injections (five-fold). The transferred donor mitochondria were stably maintained during several successive cell divisions, revealing that they were viable and functional. Several calluses were obtained from injected protoplasts without using any selection pressure. Restriction fragment length analysis of seventeen calluses, using mitochondrial DNA probes, indicated that three contained the donor “Ogura” type mitochondria. No recombinant types of mitochondria have been observed. Flow cytometric and karyotype analyses of the calluses revealed the presence of similar amount of DNA and chromosome number as those of the recipient plants of B.napus. The application of microinjection for the manipulation of cytoplasmic composition is discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00232922
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  • 10
    Electronic Resource
    Electronic Resource
    Vital DNA staining of agarose-embedded protoplasts and cell suspensions of Nicotiana plumbaginifolia (1988)
    Springer
    Plant cell reports 7 (1988), S. 489-492 
    Details
    ISSN: 1432-203X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The DNA of agarose-embedded protoplasts of Nicotiana plumbaginifolia was stained with Hoechst 33342 by immersing microscope slides, coated with immobilized protoplasts, into Erlenmeyer flasks containing consecutively dye solution, pH-correcting washing solutions and culture medium. After staining, protoplasts regenerated cell walls, started to divide and proliferated to calli. The culture system with immobilized protoplasts permits rapid change of culture media and accurate control of experimental conditions. The staining technique offers the opportunity for continuous observation of chromosomal behaviour and cell dynamics in individual plant cells. The same staining procedure was successfully applied to DNA of plant cells in suspension. Flow cytometric analysis revealed a retarding effect of the dye on the cell cycle, but within hours the cells recovered and showed their normal growth characteristics as compared to the controls.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00272738
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