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  • Brazil nut protein, grain legumes, leguminB4 promoter  (1)
  • Grain legumes  (1)
  • 1
    ISSN: 1572-9788
    Keywords: Brazil nut protein, grain legumes, leguminB4 promoter ; seed storage protein ; sulphur amino acid deficiency ; transgenic plants ; Vicia narbonensis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract In order to evaluate the possibility of enhancing the sulphur amino acids in grain legumes, we have transferred the gene for the methionine-rich 2S albumin from Brazil nut (Bertholletia excelsa) controlled by a seed-specific promoter intoVicia narbonensis. The coding region of the 2S albumin gene that has been completely synthesized was fused to the seed-specific leguminB4 promoter fromVicia faba. Transgenic lines ofV. narbonensis were generated viaAgrobacterium-mediated gene analysis of leaves, stems, roots and seeds of four R0 plants revealed that the expression of the foreign 2S albumin gene occurred in a seed-specific manner and that the foreign protein is present at levels ranging from 1% to 4.8% of total SDS-soluble seed protein. Since the current protocol for transformation ofV. narbonensis has not yet been published, a detailed description of the method is given.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1617-4623
    Keywords: Gene transfer ; Gene expression ; 2S Brazil nut albumin ; Grain legumes ; Vicia narbonensis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The coding region of the 2S albumin gene of Brazil nut (Bertholletia excelsa H.B.K.) was completely synthesized, placed under control of the cauliflower mosaic virus (CaMV) 35S promoter and inserted into the binary vector plasmid pGSGLUC1, thus giving rise to pGSGLUC1-2S. This was used for transformation of tobacco (Nicotiana tabacum L. cv. Petit Havanna) and of the grain legume Vicia narbonensis L., mediated by the supervirulent Agrobacterium tumefaciens strain EHA 101. Putative transformants were selected by screening for neomycin phosphotransferase (NPT II) and β-glucuronidase (GUS) activities. Transgenic plants were grown until flowering and fruiting occurred. The presence of the foreign gene was confirmed by Southern analysis. GUS activity was found in all organs of the regenerated transgenic tobacco and legume plants, including the seeds. In the legume, the highest expression levels of the CaMV 35S promoter-controlled 2S albumin gene were observed in leaves and roots. 2S albumin was localized in the vacuoles of leaf mesophyll cells of transgenic tobacco. The Brazil nut protein was present in the 2S fraction after gel filtration chromatography of the legume seed proteins and could be clearly identified by immunoblotting. Analysis of seeds from the R2 progenies of the legume and of transgenic tobacco plants revealed Mendelian inheritance of the foreign gene. Agrobacterium rhizogenes strain RifR 15834 harbouring the binary vector pGSGLUCl2S was also used to transform Pisum sativum L. and Vicia faba L. Hairy roots expressed the 2S albumin-specific gene. Several shoots were raised but they never completely rooted and no fertile plants were obtained from these transformants.
    Type of Medium: Electronic Resource
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