title: Combinatorial genetic transformation of cereals and the creation of metabolic libraries for the carotenoid pathway.
abstract: Combinatorial nuclear transformation is used to generate populations of transgenic plants containing random selections from a collection of input transgenes. This is a useful approach because it provides the means to test different combinations of genes without the need for separate transformation experiments, allowing the comprehensive analysis of metabolic pathways and other genetic systems requiring the coordinated expression of multiple genes. The principle of combinatorial nuclear transformation is demonstrated in this chapter through protocols developed in our laboratory that allow combinations of genes encoding enzymes in the carotenoid biosynthesis pathway to be introduced into rice and a white-endosperm variety of corn. These allow the accumulation of carotenoids to be screened initially by the colour of the endosperm, which ranges from white through various shades of yellow and orange depending on the types and quantities of carotenoids present. The protocols cover the preparation of DNA-coated metal particles, the transformation of corn and rice plants by particle bombardment, the regeneration of transgenic plants, the extraction of carotenoids from plant tissues, and their analysis by high-performance liquid chromatography.
authors.affiliation: Department of Plant Production and Forestry Science, ETSEA, University of Lleida, Lleida, Spain.
authors.forename: Gemma
authors.initials: G
authors.lastname: Farre
authors.forename: Shaista
authors.initials: S
authors.lastname: Naqvi
authors.forename: Georgina
authors.initials: G
authors.lastname: Sanahuja
authors.forename: Chao
authors.initials: C
authors.lastname: Bai
authors.forename: Uxue
authors.initials: U
authors.lastname: Zorrilla-López
authors.forename: Sol M
authors.initials: SM
authors.lastname: Rivera
authors.forename: Ramon
authors.initials: R
authors.lastname: Canela
authors.forename: Gerhard
authors.initials: G
authors.lastname: Sandman
authors.forename: Richard M
authors.initials: RM
authors.lastname: Twyman
authors.forename: Teresa
authors.initials: T
authors.lastname: Capell
authors.forename: Changfu
authors.initials: C
authors.lastname: Zhu
authors.forename: Paul
authors.initials: P
authors.lastname: Christou
mesh_terms: D000123:Acetyltransferases / Q000235:genetics; D002338:Carotenoids / Q000378:metabolism*; D002523:Edible Grain / Q000235:genetics; D018506:Gene Expression Regulation, Plant; D015723:Gene Library; D018014:Gene Transfer Techniques; D005818:Genetic Engineering; D053209:Herbicide Resistance / Q000235:genetics; D006540:Herbicides / Q000494:pharmacology; D053858:Metabolic Networks and Pathways; D012275:Oryza / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D030821:Plants, Genetically Modified; D014170:Transformation, Genetic; D003313:Zea mays / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism
chemical_list: D006540:Herbicides; D002338:Carotenoids; D000123:Acetyltransferases; C056243:phosphinothricin N-acetyltransferase