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  • 1
    Electronic Resource
    Electronic Resource
    Retention of the capacity to produce plants from protoplasts in cryopreserved cell lines of rice (Oryza sativa L.) (1991)
    Springer
    Plant cell reports 10 (1991), S. 171-174 
    Details
    ISSN: 1432-203X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A method is described for cryopreservation of cell suspension lines of rice (Oryza sativa L.) for use in protoplast research and as a way of retaining desirable characteristics of cell lines. The procedure involves pre-culture with mannitol, addition of a cryoprotectant solution of sucrose, dimethyl sulfoxide, glycerol and L-proline, two step freezing and storage in liquid nitrogen. Cells have been preserved for up to 14 months (the longest period tried in these experiments). Cryopreserved cells proliferated after plating on solid medium and new cell suspensions could be initiated within 15 days. Viable protoplasts, capable of divisions and callus formation, could be obtained 15–21 days after thawing. Variation between cell lines in terms of recovery rate after cryopreservation occurred. Differences between cell lines in plating efficiencies on solidified medium, however, contributed to this variation. Protoplasts from cryopreserved regenerable cell lines gave rise to embryogenic callus from which plants could be regenerated. These plants developed to maturity. A transformed cell line was also cryopreserved and it had retained the hygromycin resistance and regenerative capacity of the original cell line.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00234288
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  • 2
    Electronic Resource
    Electronic Resource
    RNA processing in yeast mitochondria: characterization of mit− mutants disturbed in the synthesis of subunit I of cytochrome c oxidase (1984)
    Springer
    Current genetics 8 (1984), S. 457-465 
    Details
    ISSN: 1432-0983
    Keywords: RNA processing ; Yeast mitochondria ; mit− mutants ; RNA splicing
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Mit− mutants disturbed in the synthesis of cytochrome c oxidase subunit I lack the mRNA for this protein and accumulate longer RNAs still containing intron sequences. We have analyzed the patterns of transcripts occurring in several such mutants in an attempt to define a pathway of processing events and to demarcate intron-sequences involved in RNA splicing. We find that processing does not follow a strictly ordered pathway and, in contrast to the situation for the cytochrome b gene, that a block in the processing of an intron does not necessarily lead to a block in the processing of introns downstream. Although in some cases, this may result from overlapping specificities of intronic-URF encoded RNA maturases, an internal start of translation on precursor RNAs seems more likely. M5-16, a mutant deleted for a large part of the central portion of the subunit I gene exhibits delayed processing and a highly simplified pattern of intermediates. The lengths of these indicate that maturation of the mRNA for subunit I involves processing, as well as splicing.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00433912
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  • 3
    Electronic Resource
    Electronic Resource
    Translation controls the expression level of a chimaeric reporter gene (1992)
    Springer
    Plant molecular biology 20 (1992), S. 921-938 
    Details
    ISSN: 1573-5028
    Keywords: β-D-glucuronidase ; mannopine synthase promoter ; Agrobacterium ; gene expression ; initiation of translation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transcriptional and translational fusions between the reading frame of the β-D-glucuronidase gene (gusA) and the 2′ as well as the 1′ promoter of mannopine synthase (mas), a TR locus of Agrobacterium tumefaciens, were made. The expression of these constructs was studied in the transgenic F1 offspring of independent tobacco transformants at the protein level by assaying for GUS activity and western blot analysis of the GUS protein and at the steady-state mRNA level. In leaves, stems and roots no correlation was found between steady-state levels of GUS mRNA and enzyme activity. In older tissues significantly higher GUS activities were found. This is explained by the stable character of the GUS protein together with an accumulation of protein upon ageing. Three to ten times higher GUS activities were found for in vitro grown plants than for greenhouse-grown plants of the same offspring, despite similar levels of GUS mRNA. Roots from in vitro grown plants display three to ten times higher GUS activities than stems and leaves. In transgenic plants grown in vitro, containing a translational fusion with two AUGs in phase, the initiation of translation in leaf material occurred at both AUGs. Initiation of translation at the first AUG, however, was ten times more frequent. In contrast, initiation in roots from in vitro grown plants occurred exclusively at the second AUG.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00027163
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  • 4
    Electronic Resource
    Electronic Resource
    Transgenic rice cell lines and plants: expression of transferred chimeric genes (1991)
    Springer
    Plant molecular biology 16 (1991), S. 807-820 
    Details
    ISSN: 1573-5028
    Keywords: methylation ; Oryza sativa ; protoplasts ; transformation ; β-D-glucuronidase ; methotrexate ; hygromycin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cell suspension-derived rice (Oryza sativa L.) protoplasts were transformed by direct gene uptake. PEG-mediated transformation was more efficient than electroporation. Plasmid DNA containing a hygromycin phosphotransferase (HPT) gene (which confers hygromycin resistance) driven by the CaMV 35S promoter and a β-D-glucuronidase (GUS) gene under control of the 1′, 2′ double promoter of the mannopine synthase (mas) locus of Agrobacterium tumefaciens was introduced into rice protoplasts. Southern analysis of DNA from transformed cell lines showed that the HPT and GUS genes were present intact. Both genes were expressed in transgenic cell suspensions. GUS activity was detected by histochemical staining of the cells and by enzyme assays. During a 12-day culture period the proportion of stained cells rose to a maximum and then decreased again. Considerably higher numbers of blue-stained cells were obtained when the transgenic cell lines were grown in the presence of 5-azacytidine. Transcripts of the GUS gene could not be detected, in contrast with the HPT gene. Plantlets were regenerated from one transgenic cell line. GUS activity was found in both leaf and root tissues of these plants, particularly, but not exclusively, in vascular bundles. A mouse dihydrofolate reductase coding sequence (DHFR), conferring methotrexate resistance, fused to the CaMV 35S promotor and the wild-type nopaline synthase (NOS) gene of A. tumefaciens were also introduced into rice protoplasts. Stable integration of both genes was confirmed by Southern analysis. Expression of the DHFR gene was demonstrated by high levels of resistance to methotrexate of the transgenic cell suspensions and by the presence of DHFR transcripts. Expression of the NOS gene at enzyme or RNA level was not detected. Southern analysis suggests that this gene was probably either methylated or scrambled in these lines.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00015073
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  • 5
    Electronic Resource
    Electronic Resource
    Promoters of auxin-induced genes from tobacco can lead to auxin-inducible and root tip-specific expression (1991)
    Springer
    Plant molecular biology 16 (1991), S. 983-998 
    Details
    ISSN: 1573-5028
    Keywords: auxin-induced genes ; histochemical localization ; Nicotiana tabacum ; root tip-specific expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In previous studies we have identified several mRNAs which accumulate after addition of 2,4-dichlorophenoxyacetic-acid (2,4-D) to auxin-starved tobacco cells [45, 46]. The mRNAs corresponding to cDNA clone pCNT103 were found to accumulate transiently prior to the cell division response due to auxin treatment. In this study we determined the sequences of three 103-like cDNAs and two 103-like genes, GNT1 and GNT35. To further study the regulation of the expression of these genes their 5′ regions were translationally fused with the β-D-glucuronidase reporter gene (GUS). The GNT1 5′ region led to GUS expression only in the root tips of transgenic plants. By using transgenic hairy-root cultures and transformed cell suspension cultures it was shown that the 5′ regions of both GNT1 and GNT35 lead to 2,4-D-inducible expression of GUS activity. The homology of the 103-like genes with other auxin-regulated genes is evaluated.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00016071
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  • 6
    Electronic Resource
    Electronic Resource
    Efficient plant regeneration through somatic embryogenesis from callus induced on mature rice embryos (Oryza sativa L.) (1994)
    Springer
    Plant cell, tissue and organ culture 36 (1994), S. 259-264 
    Details
    ISSN: 1573-5044
    Keywords: monocotyledons ; Oryza sativa L. ; plant regeneration ; rice ; somatic embryogenesis ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract To obtain a reproducible efficient procedure for regeneration of rice plants through somatic embryogenesis from callus four published methods of callus induction and regeneration were compared. Callus was initiated from mature embryos of the Japonica cultivar Taipei 309 of rice (Oryza sativa L.). The number, mass and morphology of the callus formed on the scutellum were dependent on the medium used. A limited humidity and an optimal aeration of the culture vessels enhanced the frequency of embryogenesis and plant regeneration. A method described by Poonsapaya et al. (1989) was found to be the most efficient and was slightly modified. As a result 98% of the T309 embryos formed callus, of which 63% regenerated into plants. Each callus yielded an average of 6 plants. Plant morphology, fertility and seed set of the regenerants were found to be normal.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00037729
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  • 7
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    RNA processing in yeast mitochondria: characterization of mit− mutants disturbed in the synthesis of subunit I of cytochrome c oxidase (1984)
    Springer
    Details
    Publication Date: 1984-08-01
    Print ISSN: 0172-8083
    Electronic ISSN: 1432-0983
    Topics: Biology
    Published by Springer
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