Girgi, MO'Kennedy, Maretha MMorgenstern, AMayer, GLorz, HOldach, KH2007-02-062007-06-072007-02-062007-06-072002Girgi, M, et al. 2002. Transgenic and herbicide resistant pearl millet (Pennisetum glaucum L.) R.Br. via microprojectile bombardment of scutellar tissue. Molecular Breeding, vol 10(4), pp 243-2521380-3743http://hdl.handle.net/10204/1560http://hdl.handle.net/10204/1560Four different pearl millet breeding lines were transformed and led to the regeneration of fertile transgenic plants. Scutellar tissue was bombarded with two plasmids containing the bar selectable marker and the beta-glucuronidase reporter gene (gus or uidA) under control of the constitutive CaMV 35S promoter or the maize Ubiquitin1 promoter (the CaMV 35S is not a maize promoter). For the delivery of the DNA-coated microprojectiles, either the particle gun PDS 1000/He or the particle inflow gun was used. The calli and regenerants were selected for their resistance to the herbicide Basta (glufosinate ammonium) mediated by the bar gene. Putative transformants were screened for enzyme activity by painting selected leaves or spraying whole plants with an aqueous solution of the herbicide Basta and by the histochemical GUS assay using cut leaf segments. PCR and Southern blot analysis of genomic DNA indicated the presence of introduced foreign genes in the genomic DNA of the transformants. Five regenerated plants represent independent transformation events and have been grown to maturity and set seed. The integration of the bar selectable and the gus reporter gene was confirmed by genomic Southern blot analysis in all five plants. All five plants had multiple integrations of both marker genes. To date, the T1 progeny of three out of four lines generated by the PDS particle gun shows co-segregating marker genes, indicating an integration of the bar and the gus gene at the same locus in the genome.12023608 bytesapplication/pdfenCopyright: 2002 Kluwer Academic PublishersTransgenic pearl millet plantsBar selectable markersGenomic DNAGenomic deoxyribonucleic reporter genesBeta-glucuronidase reporter genesGus reporter genesMicroprojectile bombardmentScutellar tissuePlant sciencesGenetic engineeringP.americanum (L) LeekeArticleGirgi, M., O'Kennedy, M. M., Morgenstern, A., Mayer, G., Lorz, H., & Oldach, K. (2002). Transgenic and herbicide resistant pearl millet (Pennisetum glaucum L.) R.Br. via microprojectile bombardment of scutellar tissue. http://hdl.handle.net/10204/1560Girgi, M, Maretha M O'Kennedy, A Morgenstern, G Mayer, H Lorz, and KH Oldach "Transgenic and herbicide resistant pearl millet (Pennisetum glaucum L.) R.Br. via microprojectile bombardment of scutellar tissue." (2002) http://hdl.handle.net/10204/1560Girgi M, O'Kennedy MM, Morgenstern A, Mayer G, Lorz H, Oldach K. Transgenic and herbicide resistant pearl millet (Pennisetum glaucum L.) R.Br. via microprojectile bombardment of scutellar tissue. 2002; http://hdl.handle.net/10204/1560.TY - Article AU - Girgi, M AU - O'Kennedy, Maretha M AU - Morgenstern, A AU - Mayer, G AU - Lorz, H AU - Oldach, KH AB - Four different pearl millet breeding lines were transformed and led to the regeneration of fertile transgenic plants. Scutellar tissue was bombarded with two plasmids containing the bar selectable marker and the beta-glucuronidase reporter gene (gus or uidA) under control of the constitutive CaMV 35S promoter or the maize Ubiquitin1 promoter (the CaMV 35S is not a maize promoter). For the delivery of the DNA-coated microprojectiles, either the particle gun PDS 1000/He or the particle inflow gun was used. The calli and regenerants were selected for their resistance to the herbicide Basta (glufosinate ammonium) mediated by the bar gene. Putative transformants were screened for enzyme activity by painting selected leaves or spraying whole plants with an aqueous solution of the herbicide Basta and by the histochemical GUS assay using cut leaf segments. PCR and Southern blot analysis of genomic DNA indicated the presence of introduced foreign genes in the genomic DNA of the transformants. Five regenerated plants represent independent transformation events and have been grown to maturity and set seed. The integration of the bar selectable and the gus reporter gene was confirmed by genomic Southern blot analysis in all five plants. All five plants had multiple integrations of both marker genes. To date, the T1 progeny of three out of four lines generated by the PDS particle gun shows co-segregating marker genes, indicating an integration of the bar and the gus gene at the same locus in the genome. DA - 2002 DB - ResearchSpace DP - CSIR KW - Transgenic pearl millet plants KW - Bar selectable markers KW - Genomic DNA KW - Genomic deoxyribonucleic reporter genes KW - Beta-glucuronidase reporter genes KW - Gus reporter genes KW - Microprojectile bombardment KW - Scutellar tissue KW - Plant sciences KW - Genetic engineering KW - P.americanum (L) Leeke LK - https://researchspace.csir.co.za PY - 2002 SM - 1380-3743 T1 - Transgenic and herbicide resistant pearl millet (Pennisetum glaucum L.) R.Br. via microprojectile bombardment of scutellar tissue TI - Transgenic and herbicide resistant pearl millet (Pennisetum glaucum L.) R.Br. via microprojectile bombardment of scutellar tissue UR - http://hdl.handle.net/10204/1560 ER -