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  • 1
    Electronic Resource
    Electronic Resource
    Transformation of Medicago sativa L. Using a Ti Plasmid Derived Vector (1991)
    Oxford, UK : Blackwell Publishing Ltd
    Plant breeding 106 (1991), S. 0 
    Details
    ISSN: 1439-0523
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: A cultivar of Medicago sativa was transformed with Agrobacterium tumefaciens strain At81 binary-vector carrying the plasmid pKan3A, with the neomycin phosphotransferase gene under the control of the mannopine biosynthesis promoter. Stem segments were infected with the bacterium and kanamycin resistant calli were obtained; plant regeneration via somatic embryogenesis and polyembryogenesis was achieved only in media devoid of the antibiotic. Genetic transformation was confirmed by the presence of the structural gene through DNA-DNA hybridization and the enzymatic assay showed its functional expression. Mesophyll protoplasts, leaf calli and S1 progeny of transformed plants grew in the presence of kanamycin (100 μgml-1). Results are discussed in relation to the use of kanamycin-resistant plants in somatic hybridization in the genus Medicago.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1439-0523.1991.tb00477.x
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  • 2
    Electronic Resource
    Electronic Resource
    Extent of RFLP variability in tetraploid populations of alfalfa, Medicago sativa (1996)
    Oxford, UK : Blackwell Publishing Ltd
    Plant breeding 115 (1996), S. 0 
    Details
    ISSN: 1439-0523
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Seven widely-cultivated alfalfa varieties and three ecotypes adapted to Central Italy were used to evaluate the extent of polymorphism in that species. Twenty plants per accession were analysed with 16 RFLP probes combined with three restriction enzymes (48 probe/enzyme combination in total) and the data were used to compute the Nei's similarity index taken as a measure of inter- and intra-population RFLP variability. The varieties were, in general, more homogeneous than the ecotypes and the cultivars ‘Adriana’ and ‘Florida’ could be differentiated more easily than the others. Few accession-specific hybridizing fragments were scored and seven populations could be distinguished from the others on the basis of significant differences in the frequencies of specific fragments. The DNA of plant populations of several sizes was bulked and the ability to detect a given fragment in pooled samples was related to the fraction of plants having that fragment among the plants forming the bulk. The results are discussed with special emphasis on the practical utilization of RFLPs for varietal identification.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1439-0523.1996.tb00883.x
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  • 3
    Electronic Resource
    Electronic Resource
    Protoplast Fusion in the Genus Medicago and Isoenzyme Analysis of Parental and Somatic Hybrid Cell Lines (1991)
    Oxford, UK : Blackwell Publishing Ltd
    Plant breeding 106 (1991), S. 0 
    Details
    ISSN: 1439-0523
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Leaf mesophyll protoplasts of Medicago arborea (2n = 32) were electrofused with cell suspension and/or callus protoplasts of Medicago sativa. Heterokaryons were identified in agarose beds by their dual fluorescein isothiocianate—chlorophyll fluorescence and their coordinates were recorded. Hybrid minicalli were manually picked up and grown first in nurse culture and then in callus induction medium. Hybrid nature of the selected calli was confirmed by isoenzyme analyses. In order to verify whether morphogenesis of somatic hybrid calli was affected by cell incompatibility, mesophyll and cell suspension protoplasts, derived from the same plant of M. sativa with high embryogenic capacity, were fused. Only callus tissues derived from mesophyll protoplasts retained the highly embryogenic character of the M. sativa genotype, while hybrid cell lines were non-morphogenic and showed isoenzyme patterns similar to tissues derived from cell suspension protoplasts. The achievement of somatic hybrid plants in the genus Medicago is discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1439-0523.1991.tb00490.x
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  • 4
    Electronic Resource
    Electronic Resource
    A Deep X-ray Survey of the PMS Population of the Upper Sco-Cen Association (1998)
    Springer
    Astrophysics and space science 261 (1998), S. 121-122 
    Details
    ISSN: 1572-946X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Physics
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1002018230818
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  • 5
    Electronic Resource
    Electronic Resource
    Plant regeneration from callus protoplasts of Onobrychis viciifolia scop. (sainfoin)
    Amsterdam : Elsevier
    Plant Science 63 (1989), S. 87-94 
    Details
    ISSN: 0168-9452
    Keywords: Onobrychis viciifolia ; bloat safe forage ; plant regeneration ; 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(89)90104-0
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  • 6
    Electronic Resource
    Electronic Resource
    A maize anthocyanin transactivator induces pigmentation in hairy roots of dicotyledonous species (1998)
    Springer
    Plant cell reports 17 (1998), S. 339-344 
    Details
    ISSN: 1432-203X
    Keywords: Key words Genetic transformation ; Reporter gene ; Agrobacterium rhizogenes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Several dicotyledonous species were infected with an Agrobacterium rhizogenes binary vector harbouring the plasmid 121.Sn which contains the maize gene Sn under the constitutive promoter CaMV35S. In maize, Sn transactivates the anthocyanin pathway in different tissues. The aim of this work was to test the efficiency of this gene to regulate the anthocyanin pathway in heterologous systems and verify its suitability as a reporter gene. The pigmentation of the hairy roots was compared with hairy roots stained for β-glucuronidase activity, which were used as a control. In two polymorphic genotypes of Lotus angustissimus, DNA integration and expression were assayed. The maize gene is competent to induce anthocyanin pigmentation in different species, but the complexity of the regulatory mechanisms of anthocyanin synthesis restricts the use of Sn as a reporter gene.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s002990050404
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  • 7
    Electronic Resource
    Electronic Resource
    Production of interspecific somatic hybrid plants in the genus Medicago through protoplast fusion (1992)
    Springer
    Theoretical and applied genetics 84 (1992), S. 792-797 
    Details
    ISSN: 1432-2242
    Keywords: M. sativa ; M. coerulea ; Protoplast electrofusion ; Somatic hybrids ; Morphological analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Symmetric somatic hybrid plants have been produced by electrofusion of leaf protoplasts of Medicago sativa and callus protoplasts of Medicago coerulea. The selection of hybrid individuals has been performed at the cellular level by recording the positions of single heterocaryons immobilized in a semisolid culture medium. The hybrid nature of the heterokaryons was assessed in fluorescent light on the basis of their color. Hybrid minicalli were picked up manually and grown first on propagating, and then on regenerating, media. Six putative hybrid calli were selected and two of them regenerated several plants. The hybrid nature of the regenerants was confirmed by cytological and isozyme analysis. Among the several morphological traits taken into account for the characterization of somatic hybrid plants, some were intermediate, some lower, and some higher, with respect to the parents. The somatic hybrid plants were fertile and set seed. The production of somatic hybrid plants in the genus Medicago is discussed in relation to the regenerating capability of parental protoplasts.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00227386
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  • 8
    Electronic Resource
    Electronic Resource
    Somatic hybrid plants between the forage legumes Medicago sativa L. and Medicago arborea L. (1996)
    Springer
    Theoretical and applied genetics 93 (1996), S. 183-189 
    Details
    ISSN: 1432-2242
    Keywords: Key wordsM. sativa ; M. arborea ; Protoplast electro-fusion ; Somatic hybrids ; Genome rearrangements
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Interspecific somatic hybrid plants were obtained by symmetrical electrofusion of mesophyll protoplasts of Medicago sativa with callus protoplasts of Medicago arborea. Somatic hybrid calli were picked manually from semi-solid culture medium after they were identified by their dual color in fluorescent light. Twelve putative hybrid calli were selected and one of them regenerated plants. The morphogenesis of the somatic hybrid calli was induced by the synthetic growth regulator 1,2 benzisoxazole-3-acetic acid. Somatic hybrid plants showed intensive genome rearrangements, as evidenced by isozyme and RFLP analysis. The morphology of somatic hybrid plants was in general intermediate between the parents. The production of hybrids by protoplast fusion between sexually incompatible Medicago species is related to the in vitro responsiveness of the parental protoplasts. The possibility of using somatic hybrid plants in alfalfa breeding is discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s001220050264
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  • 9
    Electronic Resource
    Electronic Resource
    Hairy root transformation in alfalfa (Medicago sativa L.) (1987)
    Springer
    Theoretical and applied genetics 73 (1987), S. 523-530 
    Details
    ISSN: 1432-2242
    Keywords: Hairy root ; Medicago sativa L. ; Phenotypic variation ; Agrobacterium rhizogenes ; Genetic transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The widely cultivated forage legume alfalfa (Medicago sativa L.) was transformed with the agropine type Agrobacterium rhizogenes NCPPB 1855. Sterile root and callus cultures were derived from tumorous hairy roots which were easily obtained independent of the plant variety or genotype. Plant regeneration, via somatic embryogenesis, was achieved only when a selected alfalfa line, characterized by high regenerative capability, was utilized. Genetic transformation was confirmed by the presence of agropine and T-DNA. Phenotypic alterations, mainly affecting the root system, were observed in transformed plants. The possibility that T-DNA-induced variations could be useful in the improvement of M. sativa is discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00289189
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  • 10
    Electronic Resource
    Electronic Resource
    Transcription of a maize cDNA in Lotus corniculatus is regulated by T-DNA methylation and transgene copy number (1999)
    Springer
    Theoretical and applied genetics 98 (1999), S. 257-264 
    Details
    ISSN: 1432-2242
    Keywords: Key words CaMV 35S promoter ; DNA methylation ; Gene silencing ; Lotus corniculatus ; rbcS Promoter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract   Lotus corniculatus plants transformed with a maize cDNA (G1L) encoding a sulphur-rich γ-zein were obtained by using two fusion genes: one with the CaMV 35S promoter, the other with the ribulose bisphosphate carboxylase small subunit (rbcS) promoter. The highest expression of G1L mRNA was found in plants transformed with G1L under the rbcS promoter. The steady state level of G1L mRNA in the leaves was generally directly correlated with the G1L copy number. However, due to a transcriptional block, no G1L mRNA was detected in some of the 35S-G1L multi-copy transformants. Analyses with methylation-sensitive restriction enzymes revealed that the T-DNA of the silenced 35S-G1L transformants was methylated. T-DNA copy number, G1L silencing activity, and the state of methylation were strictly correlated in primary transformants. A cross between two 35S-G1L transformed plants, one (S) with the T-DNA methylated and the other (NS) without, showed that: (1) the methylated state of T-DNA passed through meiosis; and (2) when T-DNA copies from the two parents were combined in the progeny, the unmethylated T-DNA copies of parent NS became methylated at different levels and G1L mRNA production was dependent on the degree of methylation.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s001220051066
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