Abstract
ABA-responsive element-binding factors (ABFs), as a crucial subfamily within the basic leucine zipper (bZIP) transcription factor family, exert a significant regulatory function in the process of plant responses to stress. However, the genome-wide characteristics and low-temperature response mechanisms of ABFs in cotton remain poorly understood. Herein, through genome-wide analysis across ten Gossypium species, we systematically identified 138 ABF transcription factors characterized by conserved bZIP domains. Evolutionary analysis demonstrated that segmental duplication events, coupled with purifying selection, contributed to family expansion. Cis -element profiling of promoters suggested their involvement in hormonal signaling and abiotic stress responses. Expression analysis displayed that GhABF3 expression level was significantly up-regulated under cold stress. Genetic functional analysis confirmed that GhABF3 -overexpression in A. thaliana enhanced the cold tolerance and virus-induced gene silencing (VIGS)-mediated GhABF3- silencing in cotton compromised cold resistance. Comparative transcriptomic analysis of VIGS-GhABF3 cotton plants indicated that the pathways of hormone signaling, ascorbic acid metabolism, and flavonoid biosynthesis are the significantly enriched pathways by KEGG analysis. Integrated DAP-seq and RNA-seq analyses identified GhAO1 as a direct target of GhABF3 , which was further validated by Y1H and dual-LUC assays. The contents of AsA and flavonoid were significantly reduced in both VIGS-GhABF3 and VIGS-GhAO1 cotton plants. This research conducted a systematic analysis of the evolutionary traits of the cotton ABF family, clarified the function and molecular mechanism underlying GhABF3 -mediated enhancement of low-temperature adaptability, and thus laid a robust basis for further exploration of multi-level regulatory networks and the breeding of cold-tolerant cotton varieties. • 138 ABF members with conserved bZIP domains were found in 10 Gossypium species. • GhABF3 indicated significant cold-inducible expression. • GhABF3 positively regulated the cold tolerance in Arabidopsis and cotton. • GhABF3 directly targeted to GhAO1 to participate in cold response.
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Claims 7
Claim · model-written summary
Genome-wide analysis identified 138 ABF transcription factors across ten Gossypium species.
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“through genome-wide analysis across ten Gossypium species, we systematically identified 138 ABF transcription factors characterized by conserved bZIP domains”
Claim · model-written summary
GhABF3 expression was significantly up-regulated under cold stress.
- Taxon
- Cotton
Gossypium hirsutumt0 - Treatment
- cold stresse1
- Gene
- GhABF3e1
- Outcome
- GhABF3 expression levele1
- Direction
- up-regulatede1
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Verbatim
t0“Cotton GhABF3-GhAO1 module”
e1“Expression analysis displayed that GhABF3 expression level was significantly up-regulated under cold stress”
Claim · model-written summary
GhABF3 overexpression enhanced cold tolerance in Arabidopsis.
- Taxon
- A. thaliana
Arabidopsis thalianae2 - Treatment
- GhABF3 -overexpressione2
- Gene
- GhABF3e2
- Outcome
- cold tolerancee2
- Direction
- enhancede2
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Verbatim
t0“Cotton GhABF3-GhAO1 module”
e2“GhABF3 -overexpression in A. thaliana enhanced the cold tolerance and virus-induced gene silencing (VIGS)-mediated GhABF3- silencing in cotton compromised cold resistance”
Claim · model-written summary
VIGS silencing of GhABF3 in cotton compromised cold resistance.
- Taxon
- cotton
Gossypium hirsutume2 - Treatment
- (VIGS)-mediated GhABF3- silencinge2
- Gene
- GhABF3e2
- Outcome
- cold resistancee2
- Direction
- compromisede2
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Verbatim
t0“Cotton GhABF3-GhAO1 module”
e2“GhABF3 -overexpression in A. thaliana enhanced the cold tolerance and virus-induced gene silencing (VIGS)-mediated GhABF3- silencing in cotton compromised cold resistance”
Claim · model-written summary
GhAO1 is a direct target of GhABF3.
- Taxon
- Cotton
Gossypium hirsutumt0 - Gene
- GhAO1e3
- Outcome
- direct target of GhABF3e3
- Qualifier
- validated by Y1H and dual-LUC assayse3
Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Verbatim
t0“Cotton GhABF3-GhAO1 module”
e3“Integrated DAP-seq and RNA-seq analyses identified GhAO1 as a direct target of GhABF3 , which was further validated by Y1H and dual-LUC assays”
Claim · model-written summary
VIGS silencing of GhABF3 significantly reduced ascorbic acid and flavonoid contents in cotton.
- Taxon
- cotton plants
Gossypium hirsutume4 - Treatment
- VIGS-GhABF3e4
- Outcome
- contents of AsA and flavonoide4
- Direction
- reducede4
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Verbatim
t0“Cotton GhABF3-GhAO1 module”
e4“The contents of AsA and flavonoid were significantly reduced in both VIGS-GhABF3 and VIGS-GhAO1 cotton plants”
Claim · model-written summary
Hormone signalling, ascorbic acid metabolism and flavonoid biosynthesis were significantly enriched in VIGS-GhABF3 cotton.
- Taxon
- Cotton
Gossypium hirsutumt0 - Outcome
- pathways of hormone signaling, ascorbic acid metabolism, and flavonoid biosynthesise5
Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Verbatim
t0“Cotton GhABF3-GhAO1 module”
e5“the pathways of hormone signaling, ascorbic acid metabolism, and flavonoid biosynthesis are the significantly enriched pathways by KEGG analysis”
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