An investigation of the dual mechanism of uORF-mediated regulation of GGP1 expression and the effects on drought stress in cotton.
5 affiliations
- 1School of Life Sciences, Zhengzhou University, Zhengzhou, Henan 450001, China.
- 2State Key Laboratory of Cotton Bio-Breeding and Integrated Utilization, Institute of Cotton Research of Chinese Academy of Agricultural Sciences, Anyang, Henan 455000, China.
- 3Xinjiang Key Laboratory of Cotton Genetic Improvement and Intelligent Production, Cotton Research Institute of Xinjiang Academy of Agricultural Sciences, Urumuqi, Xinjiang 830091, China.
- 4New Cultivar Innovation, New Zealand Institute for Bioeconomy Science Limited, Te Puke 3182, New Zealand.
- 5Xinjiang Key Laboratory of Crop Gene Editing and Germplasm Innovation, Institute of Western Agriculture, CAAS, Changji, Xinjiang 831100, China.
Abstract
Upstream open reading frames (uORFs) are intrinsic regulatory elements that fine-tune gene expression at the translational level without altering genomic sequences or protein structures. Here, we targeted the conserved uORF that resides in the 5'-untranslated region (5'-UTR) of a GDP-galactose phosphorylase gene (GhGGP1) in cotton (Gossypium hirsutum). Regulation of GGP1 is a key control point of plant L-ascorbic acid biosynthesis. Using a dual-luciferase reporter system, we demonstrate that this uORF represses translation of the downstream GGP1 coding sequence in a feedback manner in response to elevated ascorbate, confirming previous findings in other species. Furthermore, CRISPR/Cas9-mediated disruption of the uORFGhGGP1 generated homozygous uorfGhGGP1 mutants with unchanged transcript levels but markedly increased protein abundance and L-ascorbic acid accumulation. Under drought stress, these mutants exhibit reduced malondialdehyde levels, enhanced total antioxidant capacity, and improved drought tolerance. Mechanistically, we further show that under drought conditions, the transcription factor GhBBM directly binds to the uORF-containing cis-regulatory element, enhancing (GhGGP1) transcription and drought resistance. Together, our findings establish the uORFGhGGP1 as a dual-layer regulatory module that integrates transcriptional activation by GhBBM with translational control of the downstream protein encoding open reading frame, thereby fine-tuning drought stress responses in cotton.
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BibTeX
@article{liu2026investigation,
title = {{An investigation of the dual mechanism of uORF-mediated regulation of GGP1 expression and the effects on drought stress in cotton.}},
author = {Menghan Liu and Chao Li and Juyun Zheng and Yunpeng Cai and Peng Wang and Jiachen Yuan and Ye Wang and Mengyue Yan and Xueyuan Li and Sean Bulley and Xiaoyang Ge and Jingjing Zhan},
journal = {Plant physiology},
year = {2026},
month = oct,
doi = {10.1093/plphys/kiag693},
url = {https://europepmc.org/article/MED/42894401},
note = {CC BY},
}
RIS
TY - JOUR
TI - An investigation of the dual mechanism of uORF-mediated regulation of GGP1 expression and the effects on drought stress in cotton.
AU - Menghan Liu
AU - Chao Li
AU - Juyun Zheng
AU - Yunpeng Cai
AU - Peng Wang
AU - Jiachen Yuan
AU - Ye Wang
AU - Mengyue Yan
AU - Xueyuan Li
AU - Sean Bulley
AU - Xiaoyang Ge
AU - Jingjing Zhan
PY - 2026
DA - 2026/10/01
JO - Plant physiology
DO - 10.1093/plphys/kiag693
UR - https://europepmc.org/article/MED/42894401
AB - Upstream open reading frames (uORFs) are intrinsic regulatory elements that fine-tune gene expression at the translational level without altering genomic sequences or protein structures. Here, we targeted the conserved uORF that resides in the 5'-untranslated region (5'-UTR) of a GDP-galactose phosphorylase gene (GhGGP1) in cotton (Gossypium hirsutum). Regulation of GGP1 is a key control point of plant L-ascorbic acid biosynthesis. Using a dual-luciferase reporter system, we demonstrate that this uORF represses translation of the downstream GGP1 coding sequence in a feedback manner in response to elevated ascorbate, confirming previous findings in other species. Furthermore, CRISPR/Cas9-mediated disruption of the uORFGhGGP1 generated homozygous uorfGhGGP1 mutants with unchanged transcript levels but markedly increased protein abundance and L-ascorbic acid accumulation. Under drought stress, these mutants exhibit reduced malondialdehyde levels, enhanced total antioxidant capacity, and improved drought tolerance. Mechanistically, we further show that under drought conditions, the transcription factor GhBBM directly binds to the uORF-containing cis-regulatory element, enhancing (GhGGP1) transcription and drought resistance. Together, our findings establish the uORFGhGGP1 as a dual-layer regulatory module that integrates transcriptional activation by GhBBM with translational control of the downstream protein encoding open reading frame, thereby fine-tuning drought stress responses in cotton.
SN - 0032-0889
SN - 1532-2548
ER -