title: Genetic Transformation of Hordeum vulgare ssp. spontaneum for the Development of a Transposon-Based Insertional Mutagenesis System.
abstract: Domestication and intensive selective breeding of plants has triggered erosion of genetic diversity of important stress-related alleles. Researchers highlight the potential of using wild accessions as a gene source for improvement of cereals such as barley, which has major economic and social importance worldwide. Previously, we have successfully introduced the maize Ac/Ds transposon system for gene identification in cultivated barley. The objective of current research was to investigate the response of Hordeum vulgare ssp. spontaneum wild barley accessions in tissue culture to standardize parameters for introduction of Ac/Ds transposons through genetic transformation. We investigated the response of ten wild barley genotypes for callus induction, regenerative green callus induction and regeneration of fertile plants. The activity of exogenous Ac/Ds elements was observed through a transient assay on immature wild barley embryos/callus whereby transformed embryos/calli were identified by the expression of GUS. Transient Ds expression bombardment experiments were performed on 352 pieces of callus (3-5 mm each) or immature embryos in 4 genotypes of wild barley. The transformation frequency of putative transgenic callus lines based on transient GUS expression ranged between 72 and100 % in wild barley genotypes. This is the first report of a transformation system in H. vulgare ssp. spontaneum.
authors.affiliation: Plant Science Department, McGill University, Ste-Anne-de-Bellevue, Quebec, H9X 3V9, Canada.
authors.forename: Marie-Josée
authors.initials: MJ
authors.lastname: Cardinal
authors.affiliation: Department of Bioresource Engineering, McGill University, Ste-Anne-de-Bellevue, Quebec, H9X 3V9, Canada.
authors.forename: Rajvinder
authors.initials: R
authors.lastname: Kaur
authors.affiliation: Plant Science Department, McGill University, Ste-Anne-de-Bellevue, Quebec, H9X 3V9, Canada. jaswinder.singh@mcgill.ca.
authors.forename: Jaswinder
authors.initials: J
authors.lastname: Singh
mesh_terms: D004251:DNA Transposable Elements; D018744:DNA, Plant / Q000032:analysis; D001467:Hordeum / Q000145:classification / Q000235:genetics* / Q000254:growth & development*; D016254:Mutagenesis, Insertional / Q000379:methods*; D000069600:Plant Breeding; D046509:Tissue Culture Techniques; D014170:Transformation, Genetic
chemical_list: D004251:DNA Transposable Elements; D018744:DNA, Plant
keywords: Ac/Ds; Cereals; Green callus; Hordeum vulgare ssp. spontaneum; Tissue culture; Transformation