Work

The pVS1-VIR2 helper plasmid reduces regeneration efficiency of GRF-GIF in hexaploid wheat to obtain tri-allelic TaDOG1L4 mutations with reduced seed dormancy

Year: 2026

Type: preprint

Abstract: Wheat plays important role as a staple crop and in the supply of brewing ingredients, but its yield has been increasingly threatened by the occurrence of pre-harvest sprouting. While resistance breeding relies on a thorough understanding of seed dormancy genetics and its interaction with the environment, obtaining such knowledge has been hampered by the difficulty of working with wheat directly. DELAY OF GERMINATION 1 was cloned two decades ago as the first seed dormancy specific gene in Arabidopsis. However, its role in regulating cereal grain dormancy remains elusive. This study investigates the role of TaDOG1L4 in wheat seed dormancy using mutants generated by CRISPR-Cas9. Efficient wheat transformation using immature embryos suggests the usefulness of the GRF4-GIF1 protein chimera in shoot regeneration, while the pVS1-VIR2 helper plasmid seems to interfere with the transformation when combined with the GRF4-GIF1 vector. Phenotyping of TaDOG1L4 tri-allelic transgenic plants revealed its positive role in seed dormancy, as mutant seeds matured under both cool and warm temperature conditions showed reduced dormancy. These results suggest functional conservation of DOG1 in regulating seed dormancy.

Source: Research Square

DOI: 10.21203/rs.3.rs-10627119/v1

Authors: Renqiang Li, Muhammad Usama Hameed, Vincent Morren, Eduardo Andre Zelada Lau, Koen Geuten

Institution: KU Leuven

Language: English

Topic: Wheat and Barley Genetics and Pathology

Subfield: Plant Science

Field: Agricultural and Biological Sciences

Domain: Life Sciences

SDG: Zero hunger

Open Access status: green

Funder: KU Leuven