Project Number: 2050-21000-034-20-S
Project Type: Non-Assistance Cooperative Agreement
Start Date: Aug 1, 2018
End Date: Jul 31, 2021
Objectives: 1) Develop an efficient system for transformation of, and direct regeneration of plants from barley meristems; 2) to use the system for generating transformed barley plants; and 3) to enable subsequent use by the Wisconsin Crop Innovation Center (WCIC) and USDA Agricultural Research Service (ARS) for purposes within their institutional mandates.
Conduct research to develop and optimize a new barley meristem transformation system. Under the agreement, the WCIC will provide administrative, scientific, and technical staff to perform research and development necessary to create a technique for directly transforming and recovering fertile plants from barley meristems that will increase the efficiency and speed of the overall process compared to the existing immature embryo methodology. ARS will provide seeds of key varieties for which transformation systems are desired. It is anticipated that significant progress can be made over a two year period, as detailed below. A variety of tissue culture and transformation methodologies will be investigated to increase the likelihood of success in year 1. These methodologies will focus on determining the influence of tissue culture conditions, media formulations, agrobacterium strains, gene delivery methods, selectable markers, barley genotypes, and target tissue/cell substrates for DNA delivery and transformation. Experiments will generate data suitable for publication in peer-reviewed journals. To test the hypothesis that a direct-shooting meristem transformation method will reduce or eliminate somaclonal variation, seeds of null-segregants will be obtained and transferred to ARS for molecular and phenotypic tests to enable comparisons to embryogenic callus-derived null segregants (produced by ARS and/or WCIC). Initial experiments will deploy control vectors using visual markers and selection cassettes to rapidly optimize experimental direction and screen potential transgenic events. In year 2 experiments will be directed at optimizing transformation efficiency and using genes of interest (up to 10) to USDA-ARS researchers that will be incorporated into vectors by WCIC. Experiments will also be undertaken to determine the feasibility of gene editing. It is estimated that at the end of the first year of research there will be proof of concept that barley meristems can be transformed and during year 2, an efficient system will be developed for transformation and gene editing.