"Modern techniques in plant biotechnology" Contributions to the functional genomics of rice: 'the generation of transgenics for transposon mutagenesis in Oryza sativa ssp.japonica cv.Nipponbare' : a project-based approach

Eitas, Timothy K (A) LaMantia, Jonathan (A) (B) Martin, Jonathan (A) Longo, Chip (B) (C) Luo, Hong (B) Hu, Qian (B) Nelson-Vasichik, Kim (B) Kausch, Albert P. (B) (C)
Affiliations: (A): University of Rhode Island (B): HybriGene Inc. (C): life edu.org

Cereals include the most important food plants in the world: approximately half of the world’s population derives its caloric intake mainly from rice, and rice is also an outstanding model plant to study genomics of the cereal crops. The rice genome is estimated to contain 430 Mb and ~ 40,000 genes with a complete sequence scheduled for completion in about three to four years. If the ultimate rice genome database also relates sequence to mutant phenotypes, the biological function of as many of the genes as possible will have to be better understood. To contribute to this functional definition, a genetic, molecular and bioinformatics infrastructure will be necessary to eventually complete an extensive collection of rice lines for phenotypic functional analysis. Constructs have been developed and introduced into rice to disperse Ds transpositions throughout the rice genome where each event contains an independent, dispersed insertion of a genetically engineered Ds transposon at a defined chromosomal site. Transgenic rice is efficiently generated by Agrobacterium-mediated transformation and subsequent plant regeneration. The project-based objective of this study is to develop the necessary trangenics to establish a library of randomly dispersed Ds elements throughout the rice genome. We have used Agrobacterium based constructs to produce transgenics in the context of a two semester skills based course. All experiments have been carried out in Oryza sativa ssp. japonica cv. Nipponbare, the strain being sequenced by the IRGSP. Technology development objectives of this study include testing genetic selection strategies for the recovery of independent, dispersed transpositions of genetically engineered Ds elements with both enhancer and gene trap capabilities.