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  • 1
    Electronic Resource
    Electronic Resource
    Transgenic Plants of Lettuce ( Lactuca sativa ) Obtained Through Electroporation of Protoplasts (1989)
    [s.l.] : Nature Publishing Company
    Nature biotechnology 7 (1989), S. 503-508 
    Details
    ISSN: 1546-1696
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: [Auszug] Protoplasts from adult leaves of lettuce were electroporated in osmotically adjusted and buffered solutions containing plasmid DNA of either pCAMV CAT or pABDl. Transient CAT activity measurements and selection for kanamycin resistance showed that expression of foreign genes and stable ...
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1038/nbt0589-503
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  • 2
    Electronic Resource
    Electronic Resource
    Plant Regeneration from Mesophyll Protoplasts of Several Nicotiana Species (1979)
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 45 (1979), S. 0 
    Details
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: In a search for model systems in plant cell genetics studies mesophyll protoplasts from eleven species of Nicotiana with low chromosome number (N. acuminata, N. alata, N. glauca, N. glutinosa, N. langsdorffii, N. longiflora, N. otophora, N. paniculata, N. plumbaginifolia, N. suaveolens, N. sylvestris) were shown to divide in a liquid culture medium. Plants were recovered from calli originating from protoplasts of all these species except N. glutinosa.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1399-3054.1979.tb01702.x
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  • 3
    Electronic Resource
    Electronic Resource
    Improved flax regeneration from hypocotyls using thidiazuron as a cytokinin source (1994)
    Springer
    Plant cell reports 14 (1994), S. 120-124 
    Details
    ISSN: 1432-203X
    Keywords: Hypocotyl ; Linum usitatissimum ; Regeneration ; Thidiazuron
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The effects of thidiazuron, benzyladenine and zeatin were tested with respect to bud regeneration of different flax explants from hypocotyls, cotyledons and apices of two fibre varieties (Ariane, Viking) and one linseed variety (Antarès). These three cytokinins were tested either alone or in combination with naphthalene acetic acid, indole acetic acid or 2,4-dichlorophenoxyacetic acid. Hypocotyls were the most responsive explants. Thidiazuron was significantly the most effective followed by benzyladenine, and then zeatin, in inducing organogenesis from hypocotyl segments. The optimal thidiazuron concentration for bud regeneration from hypocotyls was 0.1–0.3 μM in combination with 0.01 μM of naphthalene acetic acid. Six days after plating, shoot initials began to appear on hypocotyl sections compared with ten to fifteen days when using benzyladenine or zeatin.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00233774
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  • 4
    Electronic Resource
    Electronic Resource
    Regeneration and characterization of plants produced from mature tobacco pollen protoplasts via gametosomatic hybridization (1995)
    Springer
    Plant cell reports 14 (1995), S. 204-209 
    Details
    ISSN: 1432-203X
    Keywords: Pollen-protoplast ; Protoplast fusion ; Gametosomatic plants ; Organelle transmission ; Nicotiana
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Mature pollen protoplasts (n) isolated from kanamycin resistant plants of Nicotiana tabacum (2n = 4x = 48) were fused with somatic mesophyll protoplasts (2n) of Nicotiana plumbaginifolia (2n = 20) to produce plants. A total of 3.6·106 mature pollen protoplasts were fused with 7·106 mesophyll protoplasts using a PEG/Ca2+ method. Mature pollen protoplasts did not divide in our culture conditions, and N. plumbaginifolia protoplasts stopped dividing when the protoplast-derived colonies were transferred to a selection medium containing paromomycine (20 mg·l-1). A total of 133 actively growing colonies were recovered on the selection medium containing kanamycin (100 mg·l-1). Plants from twenty resulting cell lines were confirmed as hybrids (17) or cybrids (3) based on leaf and floral morphology and fertility analysis. Isozyme pattern analysis confirmed the nuclear hybrid and cybrid nature, respectively, for 2 and 3 typical gametosomatic selected plants. Root tip squashes of 6 of the gametosomatic hybrid plants revealed chromosome numbers ranging from 44 to 68; the 3 selected cybrid plants had 48 chromosomes. Evidence for organelle transmission from the mesophyll partner in the gametosomatic plants is shown. From the analysis it can be concluded that the gametosomatic fusion involving mature pollen protoplasts (n) carrying a dominant selection marker can be convenient for synthesis of either hybrids or cybrids. Such gametosomatic fusion is therefore considered as a new approach towards the production of androgenetic plants with a choosen cytoplasm.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00233634
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  • 5
    Electronic Resource
    Electronic Resource
    Selection of transgenic flax plants is facilitated by spectinomycin (1996)
    Springer
    Transgenic research 5 (1996), S. 131-137 
    Details
    ISSN: 1573-9368
    Keywords: Agrobacterium tumefaciens ; hypocotyl ; gene transfer ; flax ; neomycin ; spectinomycin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Regeneration of transformed flax shoots after inoculation withAgrobacterium tumefaciens carrying a binary vector with either a neomycin phosphotransferase (nptII) gene and a β-glucuronidase (GUS) reporter gene or a spectinomycin resistance gene was examined. Hypocotyls from 4-day-old seedlings were inoculated with either of the twoA. tumefaciens strains. Selection and regeneration were achieved on a medium containing 0.1 μM thidiazuron, 0.01 μM napthalene acetic acid, 100 mgl−1 kanamycin sulphate or spectinomycin sulphate and 300 mgl−1 cefotaxime. Use of different neomycins for the selection of transformed tissues did select transformed calli but not transformed shoots either directly or via a callus phase. Selection based on spectinomycin resistance allowed the growth of transformed shoots. Transgenic shoots were rooted on a medium containing 100 mgl−1 spectinomycin sulphate. Integration of the spectinomycin resistance gene into the flax genome was confirmed by Southern blot hybridizations and spectinomycin resistance was shown to be inherited as a dominant Mendeliant trait. Therefore, spectinomycin resistance is more suitable for genetic engineering of flax than aminoglycoside resistance.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01969431
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  • 6
    Electronic Resource
    Electronic Resource
    Coat protein gene-mediated protection in Lactuca sativa against lettuce mosaic potyvirus strains (1997)
    Springer
    Molecular breeding 3 (1997), S. 75-86 
    Details
    ISSN: 1572-9788
    Keywords: pathogen-derived resistance ; transgenic plant ; virus resistance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Lettuce mosaic potyvirus (LMV) can be very destructive on lettuce crops worldwide. The LMV strain 0 (LMV-0) coat protein (CP) gene was engineered for expression in plants. It was introduced into three susceptible cultivars of Lactuca sativa using an improved procedure for transformation and regeneration of lettuce, by co-cultivation of leaf explants with Agrobacterium tumefaciens. Several transformants accumulated detectable levels of LMV CP. The R1 progeny of twelve R0 transformants (four plants per cultivar) with T-DNA integration at one single locus, was studied for protection against LMV. The progeny from five R0 transformants showed resistance to LMV-0, with the effectiveness of resistance depending on the development stage of the plants at the time of inoculation. The R1 and R2 progeny from one of these R0 transformants, Cocarde-9a, were more extensively analysed. The homozygous but not the hemizygous R1 plants displayed protection to LMV-0. The R2 progeny from one homozygous R1 plant were shown to be resistant to infection by LMV-0 and other LMV strains. As previously observed in other cases of potyvirus sequence-mediated protection, a phenomenon of recovery was observed in some plants, as well as complete resistance. However, this recovery phenotype was not always maintained, as opposed to the previous described cases, leading to a late progression of viral infection.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1009671925550
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  • 7
    Electronic Resource
    Electronic Resource
    A Lactuca universal hybridizer, and its use in creation of fertile interspecific somatic hybrids (1994)
    Springer
    Molecular genetics and genomics 245 (1994), S. 139-145 
    Details
    ISSN: 1617-4623
    Keywords: Protoplast fusion ; Albinism ; Resistance to kanamycin ; Plant regeneration ; Lactuca sp.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A Lactuca sativa cv. Ardente line heterozygous for a gene encoding resistance to kanamycin, a positive and dominant trait, was crossed with cv. Girelle, which is heterozygous for a recessive albinism marker. The resulting seeds yielded 25% albino seedlings, of which 50% were also resistant to kanamycin. Such plantlets (KR, a) grown in vitro were used for preparation of universal hybridizer protoplasts, since green buds that can develop on kanamycin containing-medium should result from fusion with any wild-type protoplast. To test the practicability of this selection scheme, we fused L. sativa KR, a protoplasts with protoplasts derived from various wild Lactuca as well as various other related species. Protoplast-derived cell colonies were selected for resistance to kanamycin at the regeneration stage. Green buds were regenerated after fusion with protoplasts of L. tatarica and of L. perennis. So far, 9 interspecific hybrid plants have been characterized morphologically. In addition, random amplified polymorphic DNA (RAPD) analysis with selected primers confirmed that these plants are indeed interspecific hybrids. Some plants are female-fertile and production of backcross progenies with L. sativa is in progress. Since many desirable traits such as resistances to viruses, bacteria and fungi (Bremia lactucae) have been characterized in wild Lactuca species, the use of somatic hybridization in breeding programmes now appears a practical possibility.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00283260
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  • 8
    Electronic Resource
    Electronic Resource
    Co-suppression of nitrate reductase host genes and transgenes in transgenic tobacco plants (1994)
    Springer
    Molecular genetics and genomics 243 (1994), S. 613-621 
    Details
    ISSN: 1617-4623
    Keywords: Co-suppression ; Transgene ; Nitrate reductase ; Tobacco ; Field trial
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Constructs carrying the entire or part of the tobacco nitrate reductase cDNA (NIA) cloned between the promoter and terminator sequences of the 35S RNA of the cauliflower mosaic virus were introduced into tobacco, in an attempt to improve nitrate assimilation. Several transgenic plants that had elevated NIA mRNA and nitrate reductase (NR) activity were obtained. In addition, a few plants that exhibited a chlorotic phenotype characteristic of NR-deficient mutants were also obtained. One of these plants contained no NIA mRNA, no NR activity and accumulated nitrate. This phenotype was therefore assumed to result from co-suppression of 35S-NIA transgenes and host NIA genes. NR-deficient plants were also found among the progeny of transformants overexpressing NIA mRNA. Genetic analyses indicated that these NR-deficient plants were homozygous for the 35S-NIA transgene, although not all homozygous plants were deficient for NR. The ratio of normal to NR-deficient plants in the progeny of homozygous plants remained constant at each generation, irrespective of the state of expression of the NIA genes (active or inactive) in the previous generation. This ratio also remained unchanged when field trials were performed in two areas of France: Versailles and Bergerac. The analysis of homozygous plants revealed that co-suppression was reversible at some stage of sexual reproduction. Indeed, host genes and transgenes reactivated at each generation, and co-suppression always appeared after a lag period of normal growth, suggesting that the phenomenon is developmentaly regulated. We observed that the triggering of cosuppression was delayed when plants were initially grown under limited light and/or watered with limited nitrate supply (light and nitrate both being required for the expression of the host NIA genes). However, this delay did not affect the final ratio between normal and NR-deficient plants after transfer to nitrate-fertilized fields. Independent transformants exhibited either different co-suppression ratios or no co-suppression at all, irrespective of the transgene copy number, suggesting that genomic sequences surrounding the transgene might play a role in determining co-suppression.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00279570
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  • 9
    Electronic Resource
    Electronic Resource
    Somatic hybrids of plants by fusion of protoplasts (1978)
    Springer
    Molecular genetics and genomics 165 (1978), S. 239-245 
    Details
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Fusions of protoplasts from Nicotiana langsdorffii and Nicotiana glauca were induced using polyethylene glycol. Parasexual hybrid colonies were selected for their ability to grow without growth substances. Hybrid plants, regenerated after grafting, were all tumorous and exhibited morphological and chromosome number variations. Out of 48 colonies selected in vitro only 6 regenerated flowering plants. Two of these plants had 42 chromosomes and were morphologically identical to the sexual amphidiploid Nicotiana glaucaxlangsdorffii.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00332522
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  • 10
    Electronic Resource
    Electronic Resource
    Sexual and somatic hybridization in the genus Lactuca (1955)
    Springer
    Euphytica 85 (1955), S. 281-285 
    Details
    ISSN: 1573-5060
    Keywords: Lactuca sativa ; Lactuca virosa ; Lactuca tatarica ; Lactuca perennis ; Iettuce ; sexual hybridization ; embryo rescue ; somatic hybridization ; protoplast fusion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Various genes for disease resistance identified in wild Lactuca are difficult, even impossible to exploit in lettuce breeding, due to sexual incompatibility between L. sativa and wild Lactuca sp. We adapted two cellular biology techniques to overcome these interspecific barriers: in vitro embryo rescue and protoplast fusion. In vitro rescue of immature embryos was used successfully for sexual hybridization between L. sativa and L. virosa. Vigorous hybrid plants were produced between L. sativa and seven accessions of L. virosa. Protoplast fusion permitted the regeneration of somatic hybrids between L. sativa and either L. tatarica or L. perennis. Hybrids between L. sativa and L. tatarica were backcrossed to L. sativa.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00023957
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