Biology subjects

Walling, J. G.

Publications and source records attributed to Walling, J. G..

2 recordsLinked to original sources

bioRxiv2026

Transposons Triggered Dynamic Evolution of MKK3 Gene, a Key Regulator for Seed Dormancy in Barley

Barley (Hordeum vulgare L.) is an important crop in the world and its seed dormancy is primarily controlled by a Mitogen-Activated Protein Kinase Kinase 3 (MKK3) gene. Although kinase activity of MKK3 and its roles in barley post-domestication have been widely studied, the pre-domestication evolution of MKK3 and the spread of nondormant alleles among global barley varieties remain largely unexplored. In this study, we analyzed MKK3 sequences in barley and its wild progenitor (H. spontaneum) and identified two polymorphic miniature inverted-repeat transposable elements (MITEs). Comparative analyses indicated that the insertions/excision of the MITEs predated the current estimates of barley domestication. Examination of the barley pangenomes coupled with droplet digital (dd) PCR revealed extensive copy number variation of MKK3 and suggested that transposons likely drove tandem amplification of the MKK3 gene on chromosome 5H. Additionally, approximately 1-Kb MKK3 sequences were found on chromosomes 1H and 6H. Further analysis indicated that these short MKK3 sequences were captured by a CACTA transposon that also contained fragments from four other expressed genes. The acquisition of MKK3 was estimated to be between 1.9-2.5 million years ago. Together, these findings illuminate the dynamic pre-domestication evolution of the MKK3 gene and suggest three independent origins of highly nondormant barley worldwide including a unique lineage predominant in Ethiopian germplasm. This study reveals the pivotal roles of transposons in MKK3 evolution and provide helpful information for understanding the complex history of MKK3 gene in barley and also for improving preharvest sprouting (PSH) tolerant varieties under distinct natural conditions.

plant biology↗

bioRxiv2025

Development and quality assessment of low-cost benchtop malting protocol for laboratory-scale malt quality evaluation

High-quality malt is influenced by three primary factors: barley genotype, environmental conditions, and malting process. To effectively evaluate malting barley breeding material and assess how environmental changes influence malt quality, it is essential to have laboratory- scale malting methods that can produce malt approximating that produced by commercial malting operations. However, existing laboratory-scale malting procedures often demand large quantities of grain, rely on specialized equipment, and are costly. To overcome these challenges, we developed a small sample-scale benchtop malting method utilizing standard laboratory equipment and components available at hardware stores. We validated the method by conducting standard malt quality tests including diastatic power, -amylase activity, total malt protein, and wort composition (soluble protein, wort soluble/total malt protein, {beta}-glucan, free amino nitrogen, and malt extract). Our findings indicate that the benchtop malting method yields quality metrics comparable to those obtained from established small-scale and full-scale malting protocols. Furthermore, a key innovation of this system is the use of separate Erlenmeyer flasks for malting each sample. Unlike conventional shared malting systems, this design enables precise measurement and comparison of treatment effects across samples malted simultaneously. This reliable, low-cost, and efficient method provides a valuable tool for screening malt quality traits in breeding lines with limited sample sizes and for testing malting regimes aimed at improving malt quality and efficiency. Additionally, it offers an accessible solution for producing high-quality, research-scale malt in laboratories without dedicated quality assurance facilities.

biochemistry↗