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  • 1
    ISSN: 1439-0523
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: The effect of regeneration of Lolium perenne and Festuca rubra from embryogenic suspension cells and protoplasts on fertility and growth was evaluated. Embryogenic suspension cultures were either routinely subcultured or cryopreserved and re-established. Phenology, morphology and fertility of regenerated plants were studied for two growing seasons in a replicated field experiment. Most regenerated L. perenne and F. rubra plants showed a delay in inflorescence emergence, a reduced seed yield and differences in morphological traits when compared with seed-grown plants. For L. perenne, performance of plants regenerated from cryopreserved suspension cultures and protoplasts was similar to that of respective plants regenerated from routinely maintained suspension cultures. However, differences in performance were observed for respective regenerants in F. rubra. The phenotypic deviation observed was partly reflected in the randomly amplified polymorphic DNA (RAPD) analysis performed. However, regenerants of both species showing similar, or even superior performance to the seed-grown plants were also found. Embryogenic suspension cells and corresponding protoplasts of L. perenne and F. rubra have the potential for producing fertile, well-performing plants which can be integrated in breeding programs.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 66 (1986), S. 0 
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Protoplasts of Brassica napus L. were cultured individually in a microdroplet system using a synthetic medium with survival rates of more than 70% and division frequencies of up to 65%. Microcallus formation occurred at frequencies of up to 50%. Factors affecting the survival and division of individually cultured protoplasts, such as composition and volume of culture medium, pH, buffering system, osmolarity and genotype, were analyzed.
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  • 3
    Electronic Resource
    Electronic Resource
    Amsterdam : Elsevier
    Plant Science 103 (1994), S. 93-106 
    ISSN: 0168-9452
    Keywords: Cryopreservation ; Embryogenic cell suspensions ; Forage grasses ; Plant regeneration from protoplasts ; Randomly amplified polymorphic DNA markers ; fescues and ryegrasses
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Amsterdam : Elsevier
    Plant Science 94 (1993), S. 179-193 
    ISSN: 0168-9452
    Keywords: Cryopreservation of suspension cultures ; Embryogenic cell suspensions ; Italian, perennial and hybrid ryegrasses ; Lolium species ; Plant regeneration from protoplasts ; Randomly amplified polymorphic DNA markers ; Somaclonal variation
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 0168-9452
    Keywords: Arabidopsis thaliana ; Catharanthus roseus ; Cytochrome P450 ; Tobacco ; Transgenic expression
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Amsterdam : Elsevier
    Plant Science 97 (1994), S. 83-94 
    ISSN: 0168-9452
    Keywords: Gene transfer to protoplasts ; Plant regeneration from protoplasts ; Red or golf course fescue (Festuca rubra) ; Suspension cultures ; Transgenic plants ; Turf and forage grasses
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1432-2145
    Keywords: Nicotiana ; Cytoplasmic male sterility ; Flower morphogenesis ; mtDNA organization ; In organello protein synthesis ; Mitochondrial transcripts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A detailed description of in planta floral ontogeny based on scanning electron microscopy, developmental histology and morphology is presented for three different alloplasmic gene-cytoplasmic male-sterile (CMS)Nicotiana tabacum isonuclear lines with cytoplasms ofN. bigelovii, N. debneyi and N. suaveolens and compared to the corresponding nuclear donorN. tabacum genotype. This allowed the precise determination of the developmental stages affected in the mutant forms as well as a thorough phenotypic characterization of them. The organization of the mitochondrial genome and expression of mitochondrial DNA (mtDNA) was investigated in the three different alloplasmic CMS tobacco analogs and compared to the corresponding malefertile parentalNicotiana species. Southern hybridizations of total cellular DNA and mtDNA from the different sets of lines with probes specific for mitochondrial genes coding for cytochrome oxidase subunits I, II and III, apocytochrome b, ATPase subunits α and 9 as well as 18S-5S ribosomal RNA indicated that: (a) mtDNA organization is different between mitochondrial genomes of fertile and sterile lines but identical in two different fertile tobacco lines; however genetic similarity among different mitochondrial genome types can be revealed by restriction fragment patterns; (b) although several differences were detected between the male-sterile and male-fertile plants, most of these were related to the origin of the mitochondria (cytoplasm donorNicotiana species); (c) identical mtDNA rearrangements — distinct to the cytoplasm donor — occur in cytoplasmic malesterile tobacco analogs bearing cytoplasm fromN. bigelovii in two differentN. tabacum nuclear backgrounds, indicating that mitochondrial genome structure inNicotiana is altered by substitution of the nuclear back-ground, since (d) inter- and intraspecific mitochondrial genome diversity among differentNicotiana species and the corresponding alloplasmic CMS tobacco analogs can be determined by hybridization with mtDNA specific probes. Analysis of in organello translation products in the three CMS-systems described confirmed the presence of variant proteins synthesized by mitochondria from CMS and male-fertileNicotiana isonuclear lines. In addition, differences due to the origin of both the nucleus and the cytoplasm, which involve changes in the presence or size of particular polypeptides, are apparent. A common feature of all three systems — including two different nuclear backgrounds — is the enhanced synthesis of a 31-kDa polypeptide in the strictly isonuclear CMS lines compared to the male-fertile tobacco. In addition, organellar gene expression was studied by Northern blot analysis of transcripts homologous to mitochondrial gene probes, revealing variant mtRNA species associated with some CMS lines.
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  • 8
    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.
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  • 9
    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.
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  • 10
    ISSN: 1432-203X
    Keywords: Forage and turf grasses ; Italian ryegrassLolium multiflorum ; Microprojectile bombardment ; Transgenic plants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transgenic forage-type Italian ryegrass (Lolium multiflorum Lam.) plants have been obtained by microprojectile bombardment of embryogenic suspension cells using a chimeric hygromycin phosphotransferase (hph) gene construct driven by riceActl 5′ regulatory sequences. Parameters for the bombardment of embryogenic suspension cultures with the particle inflow gun were partially optimized using transient expression assays of a chimericβ-glucuronidase (gusA) gene driven by the maizeUbi1 promoter. Stably transformed clones were recovered with a selection scheme using hygromycin in liquid medium followed by a plate selection. Plants were regenerated from 33% of the hygromycin-resistant calli. The transgenic nature of the regenerated plants was demonstrated by Southern hybridization analysis. Expression of the transgene in transformed adult Italian ryegrass plants was confirmed by northern analysis and a hygromycin phosphotransferase enzyme assay.
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