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  • 1
    Electronic Resource
    Electronic Resource
    Berlin, Germany : Blackwell Verlag GmbH
    Plant breeding 124 (2005), S. 0 
    ISSN: 1439-0523
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Four barley doubled haploid populations were produced by anther culture from the reciprocal crosses between two six-row barley cultivars, ‘Plaisant’ and ‘Orria’; the doubled haploid lines (DHLs) derived from each cross were subsequently assigned to weak or vigorous populations according to the weak or vigorous nature of the originating embryos. Well-formed embryos at day 25 on the induction medium were considered vigorous, whereas embryos maturing later were considered weak. The classification of vigorous and weak was closely associated with the ratio of green to albino plantlets regenerated. A random set of 25 DHLs from each of the four populations were selected for field testing in a replicated trial. Furthermore, a second set consisting of a total of 454 unreplicated DHLs from the four populations were also field assessed for grain yield. Distortion during in vitro culture may impede regeneration of a random array of microspores from a given cross, and may bias genetic estimates of specific trait/marker association in genetic studies. However, no significant differences were detected in this study among the four populations for days to heading, height, grain yield and thousand-kernel-weight when measured on the replicated trial of 100 DHLs, nor for grain yield in the second collection of 454 entries. This suggests that the likelihood of producing improved agronomic pure lines is independent of the direction of crossing and, more importantly, independent of the time when embryos matured in the induction media, at least for these particular six-row cultivars and for the anther culture method used.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Plant breeding 117 (1998), S. 0 
    ISSN: 1439-0523
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: This study was conducted with the objective of characterizing the genetic variation among a representative set of 37 barley cultivars currently grown in Spain, using restriction fragment length polymorphism (RFLP) markers. Thirty-two RFLP probes, in combination with three restriction enzymes, were used to analyse polymorphism at the molecular level. Genetic distances (GD), based on RFLP band patterns, and coancestry coefficients (f), based on pedigree records, were calculated. Of the 95 clone-enzyme combinations analysed, 71 (74.7%) were polymorphic, representing 246 RFLP patterns. A cluster analysis of GD split the sample into five distinct germplasm groups that were consistent with the history of the cultivars (winter European, spring European, CIMMYT-ICARDA materials, the single cultivar ‘Dobla’ and Spanish local materials). The Spanish group was the most distinct one and had unique alleles at markers close to major loci determining phonological adaptation. The probes which best distinguished among groups were also identified. Genetic similarity estimates were moderately consistent with f (for cultivars with complete pedigrees). The implications for integration of diversity studies into breeding programmes are discussed.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell reports 13 (1994), S. 145-148 
    ISSN: 1432-203X
    Keywords: muskmelon ; gene transfer ; Agrobacterium ; regeneration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cotyledon explants of muskmelon (Cucumis melo L., cv. Amarillo Oro) seedlings were co-cultivated with disarmed Agrobacterium tumefaciens strain LBA4404 that contained the binary vector plasmid pBI121.1. The T-DNA region of this binary vector contains the Nopaline synthase/neomycin phosphotransferase II (NPTII) chimeric gene for kanamycin resistance and the Cauliflower Mosaic Virus 35S/β-glucuronidase (GUS) chimeric gene. After infection, the cotyledon pieces were placed in induction medium containing 100 mg/l kanamycin. Putative transformed shoots were obtained, followed by the development of morphologically normal plantlets. The transgenic nature of regenerants was demonstrated by polymerase chain reaction, Southern blot analysis, plant growth on medium selective for the transgene (NPTII) and expression of the co-transformed GUS gene. Factors affecting the transformation procedure are discussed.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-203X
    Keywords: forage grasses ; Festuca pratensis ; suspension cultures ; protoplasts ; plant regeneration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Suspension cultures from mature embryo-derived compact callus were initiated in seven meadow fescue (Festuca pratensis Huds.) cultivars. Four to six months after initiation, embryogenic suspension cultures with a moderate growth rate were established from three of them (cvs. Barmondo, Belimo and Leopard). These suspension cultures showed the capacity, maintained over six months, to regenerate green plants which could be grown to maturity under greenhouse conditions. Morphogenic suspension cultures from single genotypes of three F. pratensis cultivars (cvs. Barmondo, Belimo and Leopard) yielded large numbers of protoplasts, which upon culture in agarose beads using nurse cells formed microcalli with an overall plating efficiency in the range of 10-3 to 10-4. Mature plants were reproducibly regenerated and established in soil, from such protoplasts during a period of six months. The regeneration of fertile plants from protoplasts derived from suspension cultures of meadow fescue and its implications on gene transfer technology for this species are discussed.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-203X
    Keywords: Festuca pratensis ; suspension cultures ; protoplasts ; plant regeneration ; somaclonal variation ; genetic fidelity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A cytological and molecular analysis was performed to assess the genetic uniformity and true-to-type character of plants regenerated from 20 week-old embryogenic suspension cultures of meadow fescue (Festuca pratensis Huds.), and compared to protoplastderived plants obtained from the same cell suspension. Cytological variation was not observed in a representative sample of plants regenerated directly from the embryogenic suspensions and from protoplasts isolated therefrom. Similarly, no restriction fragment length polymorphisms (RFLPs) were detected in the mitochondrial, plastid and nuclear genomes in the plants analyzed. Randomly amplified polymorphic DNA markers (RAPDs) have been used to characterise molecularly a set of mature meadow fescue plants regenerated from these in vitro cultures. RAPD markers using 18 different short oligonucleotide primers of arbitrary nucleotide sequence in combination with polymerase chain reaction (PCR) allowed the detection of pre-existing polymorphisms in the donor genotypes, but failed to reveal newly generated variation in the protoplast-derived plants compared to their equivalent suspensionculture regenerated materials. The genetic stability of meadow fescue plants regenerated from suspension cultures and protoplasts isolated therefrom and its implications on gene transfer technology for this species are discussed.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1432-203X
    Keywords: Key words Barley ; Desiccation ; Embryos ; Doubled haploids ; Microspores
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Barley microspore-derived doubled-haploid embryos have been produced in vitro. The development of embryo desiccation technology will allow long-term storage, germplasm preservation and low delivery cost. Treatment of the microspore-derived embryos was essential to induce desiccation tolerance and to arrest further development and plant regeneration. At the concentrations used, a treatment with trehalose was more efficient than with sucrose, and mannitol was harmful to the embryos. Up to 80% of the desiccated embryos produced complete green plants when transferred to regeneration medium, by the application of a 0.6 m trehalose or a 10–5 m abscisic acid treatment to the embryos in the culture induction medium. The morphology of these plants was similar to plants produced directly from non-desiccated embryos.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1432-2242
    Keywords: Asymmetric somatic hybrids ; forage grasses: fescues and ryegrasses ; Plastome-genome interaction ; Festuca arundinacea ; Lolium multiflorum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Intergeneric asymmetric somatic hybrids have been obtained by the fusion of metabolically inactivated protoplasts from embryogenic suspension cultures ofFestuca arundinacea (recipient) and protoplasts from a non-morphogenic cell suspension ofLolium multiflorum (donor) irradiated with 10, 25, 50, 100, 250 and 500 Gy of X-rays. Regenerating calli led to the recovery of genotypically and phenotypically different asymmetric somatic hybridFestulolium plants. The genome composition of the asymmetric somatic hybrid clones was characterized by quantitative dot-blot hybridizations using dispersed repetitive DNA sequences specific to tall fescue and Italian ryegrass. Data from dot-blot hybridizations using two cloned Italian ryegrass-specific sequences as probes showed that irradiation favoured a unidirectional elimination of most or part of the donor chromosomes in asymmetric somatic hybrid clones obtained from fusion experiments using donor protoplasts irradiated at doses ≤ 250 Gy. Irradiation of cells of the donor parent with 500 Gy prior to protoplast fusion produced highly asymmetric nuclear hybrids with over 80% elimination of the donor genome as well as clones showing a complete loss of donor chromosomes. Further information on the degree of asymmetry in regenerated hybrid plants was obtained from chromosomal analysis including in situ hybridizations withL. multiflorum-specific repetitive sequences. A Southern blot hybridization analysis using one chloroplast and six mitochondrial-specific probes revealed preferentially recipient-type organelles in asymmetric somatic hybrid clones obtained from fusion experiments with donor protoplasts irradiated with doses higher than 100 Gy. It is concluded that the irradiation of donor cells before fusion at different doses can be used for producing both nuclear hybrids with limited donor DNA elimination or highly asymmetric nuclear hybrid plants in an intergeneric graminaceous combination. For a wide range of radiation doses tested (25–250Gy), the degree of the species-specific genome elimination from the irradiated partner seems not to be dose dependent. A bias towards recipient-type organelles was apparent when extensive donor nuclear genome elimination occurred.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1573-5060
    Keywords: intergeneric somatic hybrids ; forage grasses ; fescue ; Festuca arundinacea ; F. rubra ; ryegrasses ; Lolium multiflorum ; L. perenne ; Alopecurus pratensis ; species-specific repetitive DNA sequences
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Intergeneric symmetric and asymmetric somatic hybrids have been obtained by fusion of metabolically inactivated protoplasts from embryogenic suspension cultures of tall fescue (Festuca arundinacea Schreb.) and unirradiated or 10–500 Gy-irradiated protoplasts from non-morphogenic cell suspensions of Italian ryegrass (Lolium multiflorum Lam.). Genotypically and phenotypically different somatic hybrid Festulolium mature flowering plants were regenerated. Species-specific sequences from F. arundinacea and L. multiflorum being dispersed and evenly-represented in the corresponding genomes were isolated and used for the molecular characterization of the nuclear make-up of the intergeneric, somatic Festulolium plants recovered. The irradiation of Italian ryegrass protoplasts with ≤250 Gy X-rays prior to fusogenic treatment favoured the unidirectional elimination of most or part of the donor chromosomes. Irradiation of L. multiflorum protoplasts with 500 Gy produced highly asymmetric (over 80% donor genome elimination) nuclear hybrids and clones showing a complete loss of donor chromosomes. The RFLP analysis of the organellar composition in symmetric and asymmetric tall fescue (+) Italian ryegrass regenerants confirmed their somatic hybrid character and revealed a bias towards recipient-type organelles when extensive donor nuclear genome elimination had occurred. Approaches aimed at improving persistence of ryegrasses based on asymmetric somatic hybridization with largely sexually-incompatible grass species (F. rubra and Alopecurus pratensis), and at transferring the cytoplasmic male sterility trait by intra- and inter-specific hybridization in L. multiflorum and L. perenne, have been undertaken.
    Type of Medium: Electronic Resource
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