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  • Agrobacterium rhizogenes  (2)
  • flow cytometry  (2)
  • 1
    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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  • 2
    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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  • 3
    Electronic Resource
    Electronic Resource
    Production of inter-specific hybrids between Limonium perezii (Stapf) Hubb. and Limonium sinuatum (L.) Mill (1998)
    Springer
    Euphytica 102 (1998), S. 109-115 
    Details
    ISSN: 1573-5060
    Keywords: chromosome number ; embryo rescue ; flow cytometry ; interspecific hybrid ; Limonium perezii ; Limonium sinuatum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Interspecific hybrids between Limonium perezii and L. sinuatum were obtained using L. perezii as the female parent. The embryos which formed were rescued by culturing 12- to 16-day-old embryos within their ovules. The ovules containing the embryos were cultured on modified B5 medium and the embryos were later excised and re-plated onto fresh medium. The embryos grew for varying lengths of time eventually reaching lengths of up to 10 mm, before ceasing growth. At this point they died. Transfer to modified MS medium containing 3 mg l-1 TDZ for 24 hours induced callus growth in the embryo derived tissues from which shoots developed. These shoots were proliferated on modified MS medium containing IBA, BAP and GA3. Shoots transferred to modified MS medium with added IBA for 6 days initiated roots which continued development after transfer of the explants to modified growth-regulator-free MS medium. After a further 28 days the plantlets were transferred to soil. The hybrid nature of one of the embryo-derived plants was determined by flow cytometry and by examination of morphological features. The mean DNA contents of 2C nuclei from L. perezii, the hybrid and L. sinuatum were 8.69 pg, 7.59 pg and 6.42 pg, respectively. Mitotic and meiotic chromosome counts from L. perezii and L. sinuatum and their hybrid revealed that they had chromosome numbers of 14, 16 and 15, respectively. Morphological analyses of leaves and flowers showed that the hybrid displayed a number of features intermediate between both parents.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1018384329409
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  • 4
    Electronic Resource
    Electronic Resource
    Interspecific hybridisation betweenLimonium perigrinum Bergius andLimonium purpuratum L. (1995)
    Springer
    Euphytica 83 (1995), S. 215-224 
    Details
    ISSN: 1573-5060
    Keywords: chromosome number ; embryo rescue ; flow cytometry ; interspecific hybridisation ; Limonium perigrinum ; Limonium purpuratum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Interspecific hybrids betweenLimonium perigrinum andL. purpuratum were obtained usingL. perigrinum as the female parent. No hybrids were produced by the reciprocal cross. Twelve- to 15-day-old embryos were rescued and cultured within their embryo sacs on modified B5 or KM medium. After two to three days the embryos were excised from their embryo sacs and re-plated on to fresh medium. When the embryo-derived plantlets had attained a length of 1 cm they were transferred to a modified MS medium containing BA and NAA for shoot proliferation. Plantlets were transferred to modified MS medium supplemented with IBA for 24 hours for root initiation then to a modified growth-regulator-free MS medium for root growth. After a further 28 days the plantlets were transferred to soil-less medium for acclimatisation. The hybrid characteristics of one of the 15 embryo-derived plants were determined by flow cytometry and by examination of morphological features. The mean DNA contents of 2C nuclei fromL. perigrinum, the hybrid andL. purpuratum were 13.98 pg, 16.81 pg and 19.37 pg, respectively. Mitotic and meiotic chromosome counts fromL. perigrinum andL. purpuratum showed that both parents and their hybrids had identical chromosome numbers (2n=24), and that the species were closely related. Morphological analyses of leaves and flowers showed that the hybrids displayed a number of features intermediate between both parents.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01678133
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