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ARS Home » Plains Area » Fargo, North Dakota » Edward T. Schafer Agricultural Research Center » Cereal Crops Improvement Research » Research » Publications at this Location » Publication #418758
Research Project: Improvement of Disease and Pest Resistance in Barley, Durum, Oat, and Wheat Using Genetics and Genomics

Location: Cereal Crops Improvement Research

Title: Genomic dissection of barley regeneration ability 1 (Bra1) and its integration with RoboSeed for genome editing with mature embryos

Author
item AWWAD, JOSEPH - Volcani Center (ARO)
item BERNSTIEN, RON - Volcani Center (ARO)
item PRUSTY, MANAS - Volcani Center (ARO)
item LIDOR, GUY - Volcani Center (ARO)
item SHARON, AMIR - Tel Aviv University
item Yang, Shengming
item FRIMAN, EYAL - Volcani Center (ARO)

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 8/5/2024
Publication Date: 10/20/2024
Citation: Awwad, J., Bernstien, R., Prusty, M., Lidor, G., Sharon, A., Yang, S., Friman, E. 2024. Genomic dissection of barley regeneration ability 1 (Bra1) and its integration with RoboSeed for genome editing with mature embryos. Meeting Abstract. Poster No. PO44.

Interpretive Summary:

Technical Abstract: CRISPR-based breeding requires cost-effective delivery systems with high penetration in the next generations. Current methods to deliver CRISPR reagents rely on a continuous supply of immature embryos as the source material for tissue culture, which are both labor and energy-consuming, mainly during the off-season. Here, we combine genetics and engineering to identify causal variation to assist the regeneration ability of mature embryos (ME) extracted from dry seeds and show the development of RoboSeed for automated extraction of ME. Genetic analysis of F2::3 ME from a cross between cv Noga and Golden Promise identified three major QTL for regeneration ability, including the barley regeneration ability 1 (Bra1) with a recessive mode of inheritance and effects of 134% between parental alleles. Furthermore, recombinant screening in the Bra1 locus in an additional F2 population identifies 63 recombinants within the QTL interval encompassing 8.9 Mbp and 390 annotated genes. Comparative transcriptome analysis of developing calli cells at four stages of tissue culture is compared to the QTL analysis to identify candidate causal genes for functional analysis. The RoboSeed ME extraction device uses machine vision algorithms for grain detection and embryo extraction. Initial semi-automated experiments show an extraction success rate of ~85%. Our combination of genetic and mechanical engineering will accelerate genome editing using a more accessible and labor-friendly material for tissue culture.