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  • 1
    Electronic Resource
    Electronic Resource
    Electroporation: Mechanism and transient expression, stable transformation and biological effects in plant protoplasts (1991)
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 81 (1991), S. 0 
    Details
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The present state of knowledge about the mechanistic and theoretical aspects of electroporation is summarized. Parameters affecting the efficiency of transient expression and stable transformation of electroporated plant protoplasts are rewieved. Biological effects of electroporation on plant protoplasts are described.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1399-3054.1991.tb02139.x
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  • 2
    Electronic Resource
    Electronic Resource
    Sonication: A new method for gene transfer to plants (1992)
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 85 (1992), S. 0 
    Details
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Sonication is a novel method for gene transfer into plant protoplasts and intact plant cells. The mode of action of ultrasound and its chemical, biochemical and physiological effects are reviewed. The state of the art of acoustic transformation is presented and possible mechanisms are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1399-3054.1992.tb04727.x
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  • 3
    Electronic Resource
    Electronic Resource
    Direct gene transfer to plant protoplasts by mild sonication (1990)
    Springer
    Plant cell reports 9 (1990), S. 207-210 
    Details
    ISSN: 1432-203X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A novel procedure employing mild sonication for transformation of plant protoplasts is described. Transient expression of a chloramphenicol acetyltransferase (CAT) gene in protoplasts of sugar beet (Beta vulgaris L.) and tobacco (Nicotiana tabacum L.) was obtained by a brief exposure of the protoplasts to 20 kHz ultrasound in the presence of plasmid DNA. Maximum levels of CAT activity were achieved by sonication for 500–900 ms at 30–70 W electric power (0.65–1.6 W/cm2 acoustic power). This reduced the viability to 15–20 % and 60 % for sugar beet and tobacco protoplasts, respectively. Up to 12 % (sugar beet) and 81 % (tobacco) of maximum transient expression could be achieved with no significant loss of viability. Protoplasts surviving exposure to ultrasound were found to have a similar long-term viability and to regenerate to micro-calli as untreated protoplasts. Plasmid DNA concentrations of 80–110 μg/ml and sucrose concentrations of 21–28 % in the sonication medium were found to be optimal for transient expression.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00232181
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  • 4
    Electronic Resource
    Electronic Resource
    A proline-rich chitinase from Beta vulgaris (1995)
    Springer
    Plant molecular biology 27 (1995), S. 211-216 
    Details
    ISSN: 1573-5028
    Keywords: Beta vulgaris ; chitinase ; proline-rich
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A gene (Chl) encoding a novel type of chitinase was isolated from Beta vulgaris. The Ch1 protein consists of an N-terminal hydrophobic prepeptide of 25 amino acids followed by a hevein-like domain of 22 amino acid residues, an unusually long proline-rich domain of 131 amino acid residues with 90 prolines, and finally a catalytic domain of 261 amino acid residues. Proteins with similar proline-rich domains are present in some other plants. The Chl gene shows a transient expression in response to fungal infection.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00019193
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  • 5
    Electronic Resource
    Electronic Resource
    Direct gene transfer to plant protoplasts by electroporation by alternating, rectangular and exponentially decaying pulses (1990)
    Springer
    Plant cell reports 8 (1990), S. 701-705 
    Details
    ISSN: 1432-203X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The efficiency of electroporation of sugar beet (Beta vulgaris L.) and tobacco (Nicotiana tabacum L.) protoplasts by alternating, rectangular and exponentially decaying pulses was studied by assaying transient expression of an introduced gene for chloramphenicol acetyltransferase. A simple device for electroporation by alternating current was constructed. The mains (220 V) were used as power supply and the pulse duration was controlled by the blow-out of a small fuse. Electroporation of sugar beet protoplasts by alternating current and exponentially decaying pulses resulted in 3–4 fold higher transient expression compared to rectangular pulses. Transient expression in tobacco protoplasts electroporated by exponentially decaying pulses was 30 % and 85 % higher than when electroporated by rectangular and alternating current pulses, respectively.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00272098
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  • 6
    Electronic Resource
    Electronic Resource
    Transformation of the endospermous legume guar (Cyamopsis tetragonoloba L.) and analysis of transgene transmission (1999)
    Springer
    Molecular breeding 5 (1999), S. 521-529 
    Details
    ISSN: 1572-9788
    Keywords: Cyamopsis tetragonoloba ; β-lactamase inhibitor ; sulbactam ; transformation ; transgene stability ; transgenic guar
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A procedure for transformation of the large-seeded endospermous legume guar (Cyamopsis tetragonoloba L.) and a study on transmission of the transgenes to offspring generations are presented. Using Agrobacterium tumefaciens with a T-DNA construct harbouring a β-glucuronidase gene (uidA) and a neomycin phosphotransferase gene (nptII), maximum transformation frequencies of cotyledonary explants were obtained using 145 mg/l kanamycin sulfate as selective agent. Carbenicillin and cefotaxime, used for the elimination of Agrobacterium after co-culture, displayed considerable toxicity to guar tissues but replacing most of these β-lactams by the non-phytotoxic β-lactamase inhibitor sulbactam as well as addition of thidiazuron and silver thiosulfate increased transformation frequencies up to 10-fold in total. The presence of the transgenes in the primary transformants was demonstrated by genomic DNA analysis of GUS-positive shoots. Chimaeric plants (5–10%) were identified by GUS analysis at the flowering stage and were discarded. Analysis of the R1 offspring from 17 independent transformants showed that in 41% of those, the uidA gene(s) was expressed and stably inherited consistent with Mendelian genetics. This was also found for the R2 and R3 generations of single copy transformants. On the other hand, a large proportion (47%) of the primary transformants gave R1 offspring in which 100% of the plants were GUS-negative. Analysis of these plants by PCR revealed that, at least, most of the transgene sequences were absent, suggesting that they had not been transmitted from the parent transformants. This occurred at similar high frequencies (40–50%) irrespective of the estimated copy number of the transgenes. Thus, major parts of the transgenes, even when present in multiple copies, displayed aberrant transmission, at a high frequency, in the process of going from the primary transformants to the first offspring generation.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1009602722423
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  • 7
    Electronic Resource
    Electronic Resource
    Relationship between promoter strength and transformation frequencies using mannose selection for the production of transgenic sugar beet (2000)
    Springer
    Molecular breeding 6 (2000), S. 207-213 
    Details
    ISSN: 1572-9788
    Keywords: Beta vulgaris ; mannose selection ; phosphomannose isomerase ; promoter strength ; transformation frequency
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The relationship between the expression level of the selectable marker gene and transformation frequency was investigated in transgenic sugar beets with five different promoters, a modified cauliflower mosaic virus 35S RNA promoter (E35S), an enhanced nopaline synthase promoter (ENOS), a modified mannopine synthase promoter (SMAS), a heat shock protein promoter (HSP80) and a chlorophyll a/b-binding protein promoter (CAB3), to drive the expression of the selectable marker gene. The selection system employed was based on the Escherichia coli phosphomannose isomerase (PMI) gene as selectable marker gene and mannose as selective agent. The selected transgenic shoots were analysed for PMI activity and the average activity for each promoter was found to be 5.9 (HSP80), 31 (SMAS), 38 (E35S), 49 (ENOS) and 61 (CAB3) mU/mg. The weakest promoter, HSP80, resulted in the lowest transformation frequency (0.30%), suggesting that this promoter was too weak to confer sufficient resistance to mannose. On the other hand, the strongest promoters, ENOS and CAB3, only gave intermediate transformation frequencies, 0.44% and 0.47% respectively, while the somewhat weaker SMAS promoter produced the highest transformation frequency, 0.89%. Thus, these data suggest that the activity of the selectable PMI gene should be above a certain threshold level; however, above this level, no simple correlation between the PMI activities, calculated as averages, and transformation frequencies could be deduced. However, extended data analysis by dividing the transgenic shoots into 4 groups according to their PMI activities (low(〈10 mU/mg), medium (10–50 mU/mg), high (50–100 mU/mg) and very high (〉100 mU/mg) expressers) revealed a significant positive correlation between the relative number of shoots having medium levels of expression and transformation frequency. This indicated that promoters that predominanthly give rise to intermediate expression levels of the selectable PMI gene result in high transformation frequencies.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1009661801680
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  • 8
    Electronic Resource
    Electronic Resource
    Nucleotide sequence of cDNA encoding the coat protein of beet yellows virus (1991)
    Springer
    Virus genes 5 (1991), S. 267-271 
    Details
    ISSN: 1572-994X
    Keywords: beet yellows virus ; viral coat protein ; K. coli expression ; closterovirus ; nucleotide sequence ; non-5′ open reading frame
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract A cDNA clone of beet yellows viral RNA expressed the viral coat protein gene inE. coli. The sequence of the 2724 nucleotide insert revealed three open reading frames, the 3′ of which was shown to be the coat protein cistron. This cistron is expressed inE. coli, in spite of there being no obvious ribosome binding site upstream.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00568976
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  • 9
    Electronic Resource
    Electronic Resource
    Analysis of mannose selection used for transformation of sugar beet (1998)
    Springer
    Molecular breeding 4 (1998), S. 111-117 
    Details
    ISSN: 1572-9788
    Keywords: Beta vulgaris ; mannose selection ; phosphomannose isomerase ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Various factors affecting mannose selection for the production of transgenic plants were studied using Agrobacterium tumefaciens-mediated transformation of sugar beet (Beta vulgaris L.) cotyledonary explants. The selection system is based on the Escherichia coli phosphomannose isomerase (PMI) gene as selectable gene and mannose as selective agent. Transformation frequencies were about 10-fold higher than for kanamycin selection but were only obtained at low selection pressures (1.0–1.5 g/l mannose) where 20–30% of the explants produced shoots. The non-transgenic shoots were eliminated during the selection procedure by a stepwise increase in the mannose concentration up to 10 g/l. Analysis of the transformed shoots showed that the PMI activity varied from 2.4 mU/mg to 350 mU/mg but the expression level was independent of the selection pressure. Complete resistance to mannose of transformed shoots was observed already at low PMI activities (7.5 mU/mg). Genomic DNA blot analysis confirmed the presence of the PMI gene in all transformants analysed. The possible mode of action of mannose selection compared to other selection methods is discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1009633809610
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