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bioRxiv · 10.1101/2025.10.10.681749

Mutations in HOMEOBOX DOMAIN-2 improve grain protein content in wheat without significantly affecting grain yield and senescence

Safdar, L. B.·Fisk, I. D.·Pasquariello, M.·Lale, A.·Searle, I. R.·Bhosale, R.·Foulkes, J.·Boden, S. A.
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Abstract

MicroRNA-resistant alleles of HOMEOBOX DOMAIN-2 (HB-2) were recently reported to improve grain protein content (GPC) in wheat by modifying vascular development in stems to increase amino acid distribution to grain. However, it is not known if these alleles increase GPC at the expense of other physiologically important traits, such as senescence and yield-component traits. Here, we evaluated the effects of mutations in HB-2 on grain yield and nitrogen (N)-use efficiency as well as senescence and plant architecture in multi-environment glasshouse and field experiments. In glasshouse experiments, the mutations in HB-2 moderately improved GPC irrespective of N supply and without accelerating senescence or decreasing grain number or weight compared to the wildtype spring wheat cv. Cadenza. The trait analysis indicated mutant plants were more tolerant of N limitation in terms of above-ground N reduction compared to the wild type. The expression of all HB-2 homeologues increased in spikes at the lemma primordium stage in response to N limitation, indicating the later GPC increase could be a result of enhanced N sink strength determined during early development, rather than processes caused by direct changes in HB-2 expression in the grain. In field experiments, the introgression of HB-2 alleles into Australian elite spring wheat cultivars significantly improved GPC without compromising grain yield. The outcomes of this study support the introduction of miRNA-resistant HB-2 alleles, with a preference to HB-A2, into commercial wheat for improving grain quality without a yield penalty.

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Safdar, L. B., Fisk, I. D., Pasquariello, M., Lale, A., Searle, I. R., Bhosale, R., Foulkes, J., Boden, S. A.. 2025-10-11. Mutations in HOMEOBOX DOMAIN-2 improve grain protein content in wheat without significantly affecting grain yield and senescence. https://doi.org/10.1101/2025.10.10.681749

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