Genetic engineering technology has emerged as a powerful tool to elucidate mechanisms of plant disease resistance and develop disease resistant plants. This technology has potential in the battle against Fusarium graminearum (Fg) in barley, especially as no resistant barley cultivars are currently available. Nevertheless, the potential of this technology is constrained by the ability to transform and regenerate gene-edited plants in vitro, with barley proving to be particularly challenging. The established protocols for producing genetically engineered barley are genotype dependent, exhibit low efficiency, and are labor-intensive. To address this issue, we established the Barley Genetic Engineering Facility for FHB Research Community since 2022. This facility, supported by the USWBSI, provides services to develop tissue culture protocols for customer-based barley cultivars and for optimizing transformation efficiency to produce transgenic barley plants with customer-provided constructs. Plant tissue culture protocols using immature scutellum explant have been developed for several barley cultivars, including the two-rowed spring barley 'Genesis' and 'Lightning,' the winter cultivar 'Thunder,' and the six-rowed barley 'Morex’. We are currently developing barley tissue culture protocols using seedling-derived leaf tissues as explants. To optimize barley transformation efficiency, we have utilized the pUBI:RUBY vector to assist screening for transformation events. We have also implemented CNQX pretreatment to improve callus induction and plant regeneration from scutellum tissues. Additionally, we have developed constructs that incorporate morphogenic genes, such as Babyboom and Plethora, which may enhance barley transformation and regeneration efficiencies. Our optimized, genotype-independent barley tissue culture and transformation protocols will aid in the production of transgenic and gene-edited barley plants for the FHB Research Community.

ACKNOWLEDGEMENT AND DISCLAIMER

This material is based upon work partially supported by the U.S. Department of Agriculture, under Project ID #FY22-GD-003. This is a cooperative project with the U.S. Wheat & Barley Scab Initiative. Any opinions, findings, conclusions, or recommendations expressed in this publication are those of the authors and do not necessarily reflect the view of the U.S. Department of Agriculture.