VIII International Symposium on Plant Bioregulation in Fruit Production, 1998
... 1983. A plant DNA minipreparation: Version II. Plant Mol. Biol. Rep., 4: 19-21. Duran-Vila, N... more ... 1983. A plant DNA minipreparation: Version II. Plant Mol. Biol. Rep., 4: 19-21. Duran-Vila, N., V. Ortega, & L. Navarro. 1989. Morphogenesis and tissue cultures of three citrus spesies. Plant Cell Tiss Org Cult., 16. 123-133. Jefferson, RA, TA Kavanagh, & MW Bevan 1987. ...
Current Plant Science and Biotechnology in Agriculture, 1999
Improvement of woody fruit species through genetic engineering will have limited applications unl... more Improvement of woody fruit species through genetic engineering will have limited applications unless tissue from mature plants of elite varieties can be readily transformed. Transformation and regeneration of these plants are usually restricted to juvenile tissues (Cervera et al. 1998a), which exhibit greater regenerative potential (Durzan 1990) and greater susceptibility to transformation than mature ones (Smith 1920). Regenerated plants from juvenile tissues will have juvenile characters and several years will be needed before horticultural and commercial traits of the transgenic plants could be evaluated. Development of transformation procedures that could bypass the juvenile stage would greatly reduce the time involved in improving woody fruit trees by genetic engineering.
Volatile organic compounds (VOCs) are secondary metabolites acting as a language for the communic... more Volatile organic compounds (VOCs) are secondary metabolites acting as a language for the communication of plants with the environment. In orange fruits, the monoterpene D-limonene accumulates at very high levels in oil glands from the peel. Drastic down-regulation of D-limonene synthase gene expression in the peel of transgenic oranges harboring a D-limonene synthase transgene in antisense (AS) configuration altered the monoterpene profile in oil glands, mainly resulting in reduced accumulation of D-limonene. This led to fruit resistance against Penicillium digitatum (Pd), Xanthomonas citri subsp. citri (Xcc) and other specialized pathogens. Here, we analyze resistance to pathogens in independent AS and empty vector (EV) lines, which have low, medium or high D-limonene concentrations and show that the level of resistance is inversely related to the accumulation of D-limonene in orange peels, thus explaining the need of high D-limonene accumulation in mature oranges in nature for the...
Methods in molecular biology (Clifton, N.J.), 2005
Transgenic plant production has been intimately connected to the beta-glucuronidase (uidA or GUS)... more Transgenic plant production has been intimately connected to the beta-glucuronidase (uidA or GUS) gene used as a reporter marker gene. The enzyme stability and the high sensitivity and amenability of the GUS assay to qualitative (histochemical assay) and to quantitative (fluorometric or spectrophotometric assay) detection are some of the reasons that explain the extensive use of uidA gene in plant genetic transformation. Methods for uidA gene detection have been thoroughly described in the literature. The aim of this chapter is to describe the basic protocols needed for GUS detection in a plant genetic transformation laboratory.
Genetic transformation of plants relies on two independent but concurrent processes: integration ... more Genetic transformation of plants relies on two independent but concurrent processes: integration of foreign DNA into plant cells and regeneration of whole plants from these transformed cells. Cell competence for regeneration and for transformation does not always fall into the same cell type/developmental stage, and this is one of the main causes of the so-called recalcitrance for transformation of certain plant species. In this study, a detailed examination of the first steps of morphogenesis from citrus explants after co-cultivation with Agrobacterium tumefaciens was performed, and an investigation into which cells and tissues are competent for regeneration and transformation was carried out. Moreover, the role of phytohormones in the co-cultivation medium as possible enhancers of gene transfer was also studied. A highly responsive citrus genotype and well-established culture conditions were used to perform a histological analysis of morphogenesis and cell competence for transform...
VIII International Symposium on Plant Bioregulation in Fruit Production, 1998
... 1983. A plant DNA minipreparation: Version II. Plant Mol. Biol. Rep., 4: 19-21. Duran-Vila, N... more ... 1983. A plant DNA minipreparation: Version II. Plant Mol. Biol. Rep., 4: 19-21. Duran-Vila, N., V. Ortega, & L. Navarro. 1989. Morphogenesis and tissue cultures of three citrus spesies. Plant Cell Tiss Org Cult., 16. 123-133. Jefferson, RA, TA Kavanagh, & MW Bevan 1987. ...
Current Plant Science and Biotechnology in Agriculture, 1999
Improvement of woody fruit species through genetic engineering will have limited applications unl... more Improvement of woody fruit species through genetic engineering will have limited applications unless tissue from mature plants of elite varieties can be readily transformed. Transformation and regeneration of these plants are usually restricted to juvenile tissues (Cervera et al. 1998a), which exhibit greater regenerative potential (Durzan 1990) and greater susceptibility to transformation than mature ones (Smith 1920). Regenerated plants from juvenile tissues will have juvenile characters and several years will be needed before horticultural and commercial traits of the transgenic plants could be evaluated. Development of transformation procedures that could bypass the juvenile stage would greatly reduce the time involved in improving woody fruit trees by genetic engineering.
Volatile organic compounds (VOCs) are secondary metabolites acting as a language for the communic... more Volatile organic compounds (VOCs) are secondary metabolites acting as a language for the communication of plants with the environment. In orange fruits, the monoterpene D-limonene accumulates at very high levels in oil glands from the peel. Drastic down-regulation of D-limonene synthase gene expression in the peel of transgenic oranges harboring a D-limonene synthase transgene in antisense (AS) configuration altered the monoterpene profile in oil glands, mainly resulting in reduced accumulation of D-limonene. This led to fruit resistance against Penicillium digitatum (Pd), Xanthomonas citri subsp. citri (Xcc) and other specialized pathogens. Here, we analyze resistance to pathogens in independent AS and empty vector (EV) lines, which have low, medium or high D-limonene concentrations and show that the level of resistance is inversely related to the accumulation of D-limonene in orange peels, thus explaining the need of high D-limonene accumulation in mature oranges in nature for the...
Methods in molecular biology (Clifton, N.J.), 2005
Transgenic plant production has been intimately connected to the beta-glucuronidase (uidA or GUS)... more Transgenic plant production has been intimately connected to the beta-glucuronidase (uidA or GUS) gene used as a reporter marker gene. The enzyme stability and the high sensitivity and amenability of the GUS assay to qualitative (histochemical assay) and to quantitative (fluorometric or spectrophotometric assay) detection are some of the reasons that explain the extensive use of uidA gene in plant genetic transformation. Methods for uidA gene detection have been thoroughly described in the literature. The aim of this chapter is to describe the basic protocols needed for GUS detection in a plant genetic transformation laboratory.
Genetic transformation of plants relies on two independent but concurrent processes: integration ... more Genetic transformation of plants relies on two independent but concurrent processes: integration of foreign DNA into plant cells and regeneration of whole plants from these transformed cells. Cell competence for regeneration and for transformation does not always fall into the same cell type/developmental stage, and this is one of the main causes of the so-called recalcitrance for transformation of certain plant species. In this study, a detailed examination of the first steps of morphogenesis from citrus explants after co-cultivation with Agrobacterium tumefaciens was performed, and an investigation into which cells and tissues are competent for regeneration and transformation was carried out. Moreover, the role of phytohormones in the co-cultivation medium as possible enhancers of gene transfer was also studied. A highly responsive citrus genotype and well-established culture conditions were used to perform a histological analysis of morphogenesis and cell competence for transform...
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