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  • 1
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    American Association for the Advancement of Science (AAAS)
    Publication Date: 1981-09-04
    Description: 〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Bates, G W -- Jackson, E Jr -- New York, N.Y. -- Science. 1981 Sep 4;213(4512):1145.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/7268425" target="_blank"〉PubMed〈/a〉
    Keywords: Catechol O-Methyltransferase/*blood ; Erythrocytes/*enzymology ; Estrone/*analogs & derivatives ; Female ; Humans ; Hydroxyestrones/*metabolism
    Print ISSN: 0036-8075
    Electronic ISSN: 1095-9203
    Topics: Biology , Chemistry and Pharmacology , Computer Science , Medicine , Natural Sciences in General , Physics
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Journal of plant growth regulation 17 (1998), S. 221-230 
    ISSN: 1435-8107
    Keywords: Key Words. Auxin—Cytokinin—Tobacco—Protoplast—Development—cdc2
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract. The roles of auxin and cytokinin in cell cycle reactivation were studied during the first 48 h of culture of mesophyll protoplasts of Nicotiana tabacum. Using hormone delay and withdrawal studies we found that auxin was required by 0–4 h of culture, whereas cytokinin was not required until hour 10–12, which is 6–10 h before S phase. Cycloheximide blocks division, indicating that protein synthesis is required. In an effort to detect a molecular response to either hormone, we examined the expression of the cell cycle marker, cdc2. Cdc2 expression was detected by 12 h of culture, coincident with the timing of the cytokinin requirement and well before the entry into S. However, cdc2 was partially induced by either auxin or cytokinin alone, suggesting that cdc2 expression is not the primary target of either hormone. Our hormone delay experiments suggest that there are separate signal transduction pathways leading from auxin and from cytokinin to reactivation of the cell cycle and that these pathways converge before S. The underlying mechanisms for these distinct pathways remain to be elucidated.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Theoretical and applied genetics 83 (1992), S. 981-986 
    ISSN: 1432-2242
    Keywords: Tobacco mosaic virus ; Nicotiana tabacum ; Kanamycin resistance ; Hypersensitive response
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The hypersensitive response of tobacco to inoculation with tobacco mosaic virus (TMV) is controlled by a single dominant gene, the N gene. As a first step in localizing and transferring the N gene, we have prepared a line of tobacco plants in which the kanamycin-resistance (Kmr) gene is closely linked to the N gene. Nicotiana tabacum plants heterozygous for the N gene were transformed to Kmr by Agrobacterium carrying pMON200. Eighty-nine independent transformed clones were regenerated and were backcrossed with nontransformed, TMV-sensitive plants. Progeny from these crosses were screened first for Kmr; then the Kmr progeny were inoculated with TMV and scored for the hypersensitive response. Of the initial 89 clones, 68 appeared to have integrated a single functional Kmr gene. Initial tests for TMV resistance indicated possible linkage between Kmr and the N gene in 11 plants. With further testing, linkage has been established for two of these plant lines. In one of these lines, the two genes were 30–40 map units apart, and evidence of somatic instability in the linkage was obtained. However, in the second line, linkage between Kmr and the N gene was tight, and recombination between the genes in this case was only 5%. Southern hybridization revealed that this plant contained only a single copy of the Kmr gene. Linkage between Kmr and the N gene in this plant line has been verified in each of two additional backcross generations.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Theoretical and applied genetics 74 (1987), S. 718-726 
    ISSN: 1432-2242
    Keywords: Gene transfer ; Asymmetric hybrids ; Kanamycin resistance ; Tobacco ; Fertility
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Mesophyll protoplasts of a kanamycin-resistant, nopaline-positive Nicotiana plumbaginifolia seed line were inactivated by γ-irradiation and electrically fused with unirradiated mesophyll protoplasts of N. tabacum. Hybrids were selected on kanamycin and regenerated. Genetic material from N. plumbaginifolia was detected in these plants by the following criteria: (1) morphology, (2) esterase isozyme profiles, and (3) the presence of nopaline in leaf extracts. All of the plants regenerated were morphologically more similar to N. tabacum than to N. plumbaginifolia, and many were indistinguishable from N. tabacum. It was found that 37 plants displayed one or two esterases characteristic of N. plumbaginifolia in addition to a full set of esterases from N. tabacum. Based on their esterases, we have classified these plants as somatic hybrids. However, irradiation has clearly reduced the amount of N. plumbaginifolia genetic material that they retain; 24 plants were found that had only N. tabacum esterases but that produced nopaline and were kanamycin resistant. Genomic DNA from several of these plants was probed by Southern blotting for the presence of the authentic neomycin phosphotransferase gene (kanamycin-resistance gene) — all were found to contain the gene. These plants were classified as asymmetric hybrids. Finally, 25 plants were regenerated that were kanamycin sensitive, negative for nopaline, and contained only N. tabacum esterases. All of the regenerated plants, including this final category, were male sterile. As transferring the N. plumbaginifolia cytoplasm to an N. tabacum nuclear background results in an alloplasmic form of male sterility, all of the plants regenerated in this study appear to be cybrids irrespective of their nuclear constitution. Chromosome analysis of the asymmetric hybrids showed that most of them contained one more chromosome than is normal for N. tabacum. The somatic hybrids examined all had several additional chromosomes. Although male sterile, the asymmetric hybrids were female fertile to varying degrees and were successfully backcrossed with N. tabacum. Analysis of the resultant F1 progeny indicated that the kanamycin-resistance gene from N. plumbaginifolia is partially unstable during meiosis, as would be expected for factors inherited on an unpaired chromosome.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Theoretical and applied genetics 80 (1990), S. 481-487 
    ISSN: 1432-2242
    Keywords: Somatic hybridization ; Gene transfer ; Chromosome elimination ; Tobacco ; TMV resistance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Genetically asymmetric hybrids were recovered by fusion of Nicotiana tabacum protoplasts with irradiated protoplasts of kanamycin-resistant, nopalineproducing plants of N. repanda. Hybrid calli were selected by culture on media containing kanamycin and were regenerated. These plants were morphologically similar to N. tabacum but produced nopaline, indicating they retained genes from N. repanda. Esterase isozyme profiles also indicated that the plants are somatic hybrids, but are more similar to N. tabacum than N. repanda. Chromosome counts showed most of the hybrids had 55–62 chromosomes, which is consistent with extensive, although incomplete elimination of N. repanda chromosomes. The hybrids were largely male sterile, but about half of them set seed when crossed with N. tabacum. Chromosome numbers of the progeny and the pattern of inheritance of kanamycin resistance indicated the continued elimination of N. repanda genetic material in these backcrosses. The N. repanda parent used in these fusions gave a hypersensitive response to TMV, whereas the N. tabacum parent was TMV sensitive. When inoculated with TMV, plants from two hybrid clones gave a hypersensitive response. Plants from the other clones became systemically infected with the virus.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Theoretical and applied genetics 88 (1994), S. 965-972 
    ISSN: 1432-2242
    Keywords: Protoplast fusion ; Irradiation Asymmetric somatic hybrids ; Chromosome elimination Nicotiana
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Mesophyll protoplasts of a kanamycin-resistant line of Nicotiana plumbaginifolia were gamma-irradiated and fused with mesophyll protoplasts of N. tabacum plants bearing the sulfur mutation. Hybrid calli were recovered by selection on media containing kanamycin. In one group of experiments, the degree of elimination of donor (N. plumbaginifolia) genetic material in the hybrid calli was assessed by dot-blot hybridization using a N. plumbaginifolia-specific repetitive-DNA sequence as a probe. The elimination of donor DNA was found to increase with increasing gamma dose for all doses tested (5–50 krad). Elimination of donor DNA was also found to continue in the calli for the first 12 months in culture. The degree of chromosome elimination was quite variable; for a 50-krad dose, some hybrids were recovered that retained less than 15% of the donor genome, whereas others retained nearly 50%. In a second set of experiments, the degree of donorchromosome elimination was assessed from the fraction of hybrid calli that exhibited complementation of the Su phenotype due to retention of a wild-type Su allele of the donor. When N. plumbaginifolia protoplasts were inactivated by treatment with iodoacetate, rather than gamma irradiation, all the hybrid calli were green. However, when the donor protoplasts were inactivated by irradiation, the fraction of hybrid calli that were able to complement the Su mutation decreased with increasing gamma dose; for a 50-krad dose only 40% of the hybrid calli were green. From these data, the degree of radiation-induced donor-chromosome elimination was calculated and was found to agree closely with that measured by dot-blot hybridization. We conclude that radiation-induced elimination of donor chromosomes increases with gamma dose and time in culture in N. tabacum (+)N. plumbaginifolia hybrids, but that donor-chromosome elimination is an inherently variable process.
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Theoretical and applied genetics 70 (1985), S. 227-233 
    ISSN: 1432-2242
    Keywords: Electrical fusion ; Protoplast regeneration ; Isozymes ; Nicotiana
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Electrically-induced protoplast fusion has been used to produce somatic hybrids between Nicotiana plumbaginifolia and Nicotiana tabacum. Following fusion of suspension culture protoplasts (N. plumbaginifolia) with mesophyll protoplasts (N. tabacum) heterokaryons were identified visually and their development was followed in culture. Because electrical fusion is a microtechnique, procedures were developed for culturing the heterokaryons in small numbers and at low density. The fusion and culture procedures described are rapid, uncomplicated and repeatable. Good cell viabilities indicate that the fusion procedure is not cytotoxic. Fused material was cultured 1–2 days at high density in modified K3 medium (Nagy and Maliga 1976). The heterokaryons were isolated manually and grown, at low density in conditioned media. Calli have been regenerated. Esterase isozyme patterns confirm the hybrid character of calli and clonally-derived plantlets recovered from these fusions.
    Type of Medium: Electronic Resource
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  • 8
    Publication Date: 1986-08-01
    Print ISSN: 0027-8424
    Electronic ISSN: 1091-6490
    Topics: Biology , Medicine , Natural Sciences in General
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