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  • 1
    Electronic Resource
    Electronic Resource
    Regeneration of transgenic vegetable brassicas (Brassica oleracea and B. campestris) via Ri-mediated transformation (1997)
    Springer
    Plant cell reports 16 (1997), S. 587-593 
    Details
    ISSN: 1432-203X
    Keywords: Key wordsAgrobacterium rhizogenes ; Brassica oleracea ; Brassica campestris ; Transformation ; Vegetable brassicas
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A procedure for the production of fertile transgenic brassicas via Ri-mediated transformation is reported in this paper. Transgenic hairy root lines were selected for 12 vegetable brassica cultivars and lines representing six varieties: broccoli, Brussels sprouts, cabbage, cauliflower, rapid-cycling (all Brassica oleracea) and Chinese cabbage (B. campestris). Leaf explants or petioles of intact cotyledons were co-cultivated with Agrobacterium strain A4T harbouring various binary vectors. The T-DNA region of all binary vectors contained a neomycin phosphotransferase II gene for kanamycin resistance, in addition to other genes. Hairy root lines grew prolifically on hormone-free medium containing kanamycin. Transgenic shoots were regenerated from all cultivars either spontaneously or after transfer of hairy roots to a hormone-containing medium. Southern analysis confirmed that the plants were transgenic. Plants from all brassica types were successfully transferred to greenhouse conditions. Plants were fertile and segregation analysis confirmed transmission of traits to progeny.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s002990050284
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  • 2
    Electronic Resource
    Electronic Resource
    Regeneration of transgenic vegetable brassicas (Brassica oleracea andB. campestris) via Ri-mediated transformation (1997)
    Springer
    Plant cell reports 16 (1997), S. 587-593 
    Details
    ISSN: 1432-203X
    Keywords: Agrobacterium rhizogenes ; Brassica oleracea ; Brassica campestris ; Transformation ; Vegetable brassicas
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A procedure for the production of fertile transgenic brassicas via Ri-mediated transformation is reported in this paper. Transgenic hairy root lines were selected for 12 vegetable brassica cultivars and lines representing six varieties: broccoli, Brussels sprouts, cabbage, cauliflower, rapid-cycling (allBrassica oleracea) and Chinese cabbage (B. campestris). Leaf explants or petioles of intact cotyledons were co-cultivated withAgrobacterium strain A4T harbouring various binary vectors. The T-DNA region of all binary vectors contained a neomycin phosphotransferase II gene for kanamycin resistance, in addition to other genes. Hairy root lines grew prolifically on hormone-free medium containing kanamycin. Transgenic shoots were regenerated from all cultivars either spontaneously or after transfer of hairy roots to a hormone-containing medium. Southern analysis confirmed that the plants were transgenic. Plants from all brassica types were successfully transferred to greenhouse conditions. Plants were fertile and segregation analysis confirmed transmission of traits to progeny.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01275497
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    Paper (German National Licenses)
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  • 3
    Electronic Resource
    Electronic Resource
    Hairy roots of Brassica napus: II. Glutamine synthetase overexpression alters ammonia assimilation and the response to phosphinothricin (1994)
    Springer
    Plant cell reports 14 (1994), S. 41-46 
    Details
    ISSN: 1432-203X
    Keywords: Agrobacterium rhizogenes ; Brassica napus ; glutamine synthetase ; phosphinothricin ; rape ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Hairy roots of Brassica napus (rape cv. Giant) have been produced that contain the cytosolic glutamine synthetase (GS) gene from Glycine max (soybean). Leaf explants were cocultivated with Agrobacterium rhizogenes strain A4T harbouring the binary vector pLN16. This vector was constructed by inserting a soybean cytosolic GS cDNA into the multiple cloning site of pGA643, placing it under the control of the CaMV promoter. In addition, the T-DNA region of pLN16 contained a NPTII gene for selection of transformed cells. Transgenic hairy roots grew prolifically on hormone-free media containing a selective level of kanamycin. Southern and northern analyses confirmed the presence of soybean GS DNA and transcripts, respectively. These transformed hairy roots also have a greater abundance of the GS polypeptide, approximately 3–6 fold greater GS activity and lower levels of endogenous ammonia. Hairy roots provide a useful system for studying responses to phosphinothricin (PPT). Hairy roots grown in media containing PPT had lower GS activity, greater ammonia accumulation and slower growth than controls. The presence of the soybean GS gene in the hairy roots reduced these PPT-induced effects and resulted in higher GS activity, lower ammonia levels and faster growth than in PPT-treated controls. Greater tolerance of PPT was also seen in shoots regenerated from the hairy roots displaying elevated levels of GS activity.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00233296
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  • 4
    Electronic Resource
    Electronic Resource
    Atrazine-resistant cytoplasmic male-sterile-nigra broccoli obtained by protoplast fusion between cytoplasmic male-sterile Brassica oleracea and atrazine-resistant Brassica campestris (1991)
    Springer
    Theoretical and applied genetics 83 (1991), S. 201-208 
    Details
    ISSN: 1432-2242
    Keywords: Brassica ; Atrazine resistance ; Cytoplasmic male sterility ; Protoplast fusion ; CMS-nigra
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Protoplast fusion was used to combine the cytoplasmic traits of atrazine resistance and male sterility in Brassica oleracea var. italica (broccoli). Leaf protoplasts from broccoli with the petaloid B. nigra type of cytoplasmic male sterility were fused with hypocotyl protoplasts from an atrazine-resistant biotype of B. campestris var. oleifera cv Candle (oilseed rape). A total of 19 colonies regenerated shoots, all of which were broccolilike in phenotype, i.e., lacked trichomes. Four shoots, all from one colony, were atrazine resistant, surviving and growing in the presence of 25 μM atrazine. A leaf piece assay also confirmed that they were atrazine resistant. Molecular analysis showed that they contain chloroplasts from the atrazine-resistant B. campestris parent and mitochondria from the B. nigra parent. No recombination or rearrangement of the mitochondrial genomes in the fusion products was detected. These four plants and their progeny all showed the petaloid B. nigra type of male sterility.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00226252
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  • 5
    Electronic Resource
    Electronic Resource
    Rearrangement, amplification, and assortment of mitochondrial DNA molecules in cultured cells of Brassica campestris (1989)
    Springer
    Theoretical and applied genetics 77 (1989), S. 17-25 
    Details
    ISSN: 1432-2242
    Keywords: Brassica ; Callus culture ; Mitochondrial DNA ; Genomic variability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We compared Brassica campestris mitochondrial and chloroplast DNAs from whole plants and from a 2-year-old cell culture. No differences were observed in the chloroplast DNAs (cpDNAs), whereas the culture mitochondrial DNA (mtDNA) was extensively altered. Hybridization analysis revealed that the alterations are due entirely to rearrangement. At least two inversions and one large duplication are found in the culture mtDNA. The duplication element is shown to have the usual properties of a plant mtDNA high frequency “recombination repeat”. The culture mtDNA exists as a complex heterogeneous population of rearranged and unrearranged molecules. Some of the culture-associated rearranged molecules are present in low levels in native plant tissue and appear to have sorted out and amplified in the culture. Other mtDNA rearrangements may have occurred de novo. In addition to alterations of the main mitochondrial genome, an 11.3 kb linear mtDNA plasmid present in whole plants is absent from the culture. Contrary to findings in cultured cells of other plants, small circular mtDNA molecules were not detected in the B. campestris cell culture.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00292310
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  • 6
    Electronic Resource
    Electronic Resource
    Hairy roots of Brassica napus: I. Applied glutamine overcomes the effect of phosphinothricin treatment (1994)
    Springer
    Plant cell reports 14 (1994), S. 37-40 
    Details
    ISSN: 1432-203X
    Keywords: Ammonia ; Agrobacterium rhizogenes ; Brassica napus ; glutamine ; glutamine synthetase ; phosphinothricin ; rape
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Hairy roots of Brassica napus (rape cv. Giant) were produced by cocultivating leaf and cotyledon explants with Agrobacterium rhizogenes strain A4T. The hairy roots grew prolifically on solid and in liquid media. Incorporation of ammonium sulphate or phosphinothricin (PPT) into the media reduced growth. PPT treatment reduced glutamine synthetase (GS) activity and increased the ammonia content of the hairy roots. We have found that PPT treatment also induces a loss of glutamine from the roots and this may influence root growth. To test this we grew hairy roots in a liquid medium containing 10 mM glutamine. This glutamine treatment overcame the PPT induced suppression of growth but also significantly increased GS activity, reduced ammonia accumulation and increased the levels of glutamate and asparagine.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00233295
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  • 7
    Electronic Resource
    Electronic Resource
    Factors that influence Agrobacterium rhizogenes-mediated transformation of broccoli (Brassica oleracea L. var. italica) (2000)
    Springer
    Plant cell reports 19 (2000), S. 994-999 
    Details
    ISSN: 1432-203X
    Keywords: Key words Agrobacterium rhizogenes ; Brassica oleracea ; Transformation ; Arginine ; Acetosyringone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  An improved broccoli transformation system was developed by optimising several factors that affect the rate of effective Agrobacterium-mediated transformation. Leaf explants of cultivar Shogun were co-cultivated with Agrobacterium rhizogenes strain A4T harbouring the binary vector pART278. The T-DNA of this binary vector contains a neomycin phosphotransferase II (NOS-NPTII-NOS) gene for kanamycin resistance and a β-glucuronidase (35S-GUS-OCS) gene. Several media and factors were evaluated including combinations of arginine, mannopine, acetosyringone and the use of feeder cell layers. The new protocol includes the use of 200 μm acetosyringone in LB medium for bacterial growth, the use of a Brassica campestris feeder cell layer, 10 mm mannopine and 50 μm acetosyringone in the co-cultivation medium and 1 mm arginine in the selection medium. The use of this optimised protocol produced transformation rates of 33% in preliminary experiments transforming broccoli with the antisense 1-aminocyclopropane-1-carboxylic acid (ACC) oxidase gene from pTOM13.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s002990000221
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