Abstract
Chromatin remodelers are essential for regulating plant growth, development, and responses to environmental stresses. HIT4 (HEAT-INTOLERANT 4) is a novel stress-induced chromatin remodeling factor that has been less studied in abiotic stress and stress resistance, particularly in cotton. In this study, we conducted a comprehensive analysis of the members of the HIT4 gene family in Gossypium hirsutum using bioinformatics methods, including phylogenetic relationships, gene organization, transcription profiles, phylogenetic connections, selection pressure, and stress response. A total of 18 HIT4 genes were identified in four cotton species, with six HIT4 gene members in upland cotton. Based on the evolutionary relationships shown in the phylogenetic tree, the 18 HIT4 protein sequences were classified into four distinct subgroups. Furthermore, we conducted chromosome mapping to determine the genomic locations of these genes and visually represented the structural characteristics of HIT4 in G. hirsutum. In addition, we predicted the regulatory elements in HIT4 in G. hirsutum and conducted an analysis of repetitive sequences and gene collinearity among HIT4 in four cotton species. Moreover, we calculated the Ka/Ks ratio for homologous genes to assess the selection pressure acting on HIT4. Using RNA-seq, we explored the expression patterns of HIT4 genes in G. hirsutum and Gossypium barbadense. Through weighted gene co-expression network analysis (WGCNA), we found that GHHIT4_4 belonged to the MEblue module, which was mainly enriched in pathways such as DNA replication, phagosome, pentose and glucuronate interconversions, steroid biosynthesis, and starch and sucrose metabolism. This module may regulate the mechanism of upland cotton resistance to Verticillium wilt through DNA replication, phagosome, and various metabolic pathways. In addition, we performed heterologous overexpression of GH_D11G0591 (GHHIT4_4) in tobacco, and the results showed a significant reduction in disease index compared to the wild type, with higher expression levels of disease resistance genes in the transgenic tobacco. After conducting a VIGS (virus-induced gene silencing) experiment in cotton, the results indicated that silencing GHHIT4_4 had a significant impact, the resistance to Verticillium wilt weakened, and the internode length of the plants significantly decreased by 30.7% while the number of true leaves increased by 41.5%. qRT-PCR analysis indicated that GHHIT4_4 mainly enhanced cotton resistance to Verticillium wilt by indirectly regulating the PAL, 4CL, and CHI genes. The subcellular localization results revealed that GHHIT4_4 was predominantly distributed in the mitochondria and nucleus. This study offers preliminary evidence for the involvement of the GHHIT4_4 in cotton resistance to Verticillium wilt and lays the foundation for further research on the disease resistance mechanism of this gene in cotton.
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Claims 14
Claim · model-written summary
Eighteen HIT4 genes were identified across four cotton species.
- Taxon
- cotton species
Gossypium hirsutume0 - Gene
- HIT4e0
- Outcome
- HIT4 genese0
- Value
- 18e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“A total of 18 HIT4 genes were identified in four cotton species, with six HIT4 gene members in upland cotton.”
Claim · model-written summary
Six HIT4 members were identified in upland cotton.
Verbatim
e0“A total of 18 HIT4 genes were identified in four cotton species, with six HIT4 gene members in upland cotton.”
Claim · model-written summary
HIT4 protein sequences were classified into four subgroups.
- Taxon
- Gossypium hirsutumGossypium hirsutumt0
- Gene
- HIT4e0
- Outcome
- protein sequences were classified into four distinct subgroupse0
- Value
- foure0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“the 18 HIT4 protein sequences were classified into four distinct subgroups.”
t0“Gossypium hirsutum”
Claim · model-written summary
GHHIT4_4 belonged to the MEblue module.
- Taxon
- Gossypium hirsutumGossypium hirsutumt0
- Gene
- GHHIT4_4e0
- Outcome
- belonged to the MEblue modulee0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“we found that GHHIT4_4 belonged to the MEblue module, which was mainly enriched in pathways such as DNA replication, phagosome, pentose and glucuronate interconversions, steroid biosynthesis, and starch and sucrose metabolism.”
t0“Gossypium hirsutum”
Claim · model-written summary
The MEblue module was enriched in DNA replication, phagosome, and pentose and glucuronate pathways.
- Taxon
- Gossypium hirsutumGossypium hirsutumt0
- Outcome
- enriched in pathways such as DNA replication, phagosome, pentose and glucuronate interconversionse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“which was mainly enriched in pathways such as DNA replication, phagosome, pentose and glucuronate interconversions, steroid biosynthesis, and starch and sucrose metabolism.”
t0“Gossypium hirsutum”
Claim · model-written summary
The MEblue module was enriched in steroid biosynthesis and starch and sucrose metabolism.
- Taxon
- Gossypium hirsutumGossypium hirsutumt0
- Outcome
- steroid biosynthesis, and starch and sucrose metabolisme0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“which was mainly enriched in pathways such as DNA replication, phagosome, pentose and glucuronate interconversions, steroid biosynthesis, and starch and sucrose metabolism.”
t0“Gossypium hirsutum”
Claim · model-written summary
The authors propose that the module may regulate upland cotton wilt resistance through replication and metabolic pathways.
- Taxon
- upland cottone0
- Interactor
- VerticilliumVerticilliume0pathogen
- Outcome
- resistance to Verticillium wilte0
- Qualifier
- through DNA replication, phagosome, and various metabolic pathwayse0
Verbatim
e0“This module may regulate the mechanism of upland cotton resistance to Verticillium wilt through DNA replication, phagosome, and various metabolic pathways.”
Claim · model-written summary
GHHIT4_4 overexpression in tobacco significantly reduced disease index versus wild type.
- Taxon
- tobacco
Nicotiana tabacume0 - Treatment
- heterologous overexpression of GH_D11G0591 (GHHIT4_4)e0
- Gene
- GHHIT4_4e0
- Outcome
- disease indexe0
- Direction
- reductione0
- Comparator
- wild typee0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“we performed heterologous overexpression of GH_D11G0591 (GHHIT4_4) in tobacco, and the results showed a significant reduction in disease index compared to the wild type, with higher expression levels of disease resistance genes in the transgenic tobacco.”
Claim · model-written summary
GHHIT4_4 overexpression in tobacco raised expression of resistance genes versus wild type.
- Taxon
- tobaccoNicotiana tabacume0
- Treatment
- heterologous overexpression of GH_D11G0591 (GHHIT4_4)e0
- Gene
- GHHIT4_4e0
- Outcome
- expression levels of disease resistance genese0
- Direction
- highere0
- Comparator
- wild typee0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“we performed heterologous overexpression of GH_D11G0591 (GHHIT4_4) in tobacco, and the results showed a significant reduction in disease index compared to the wild type, with higher expression levels of disease resistance genes in the transgenic tobacco.”
Claim · model-written summary
Silencing GHHIT4_4 weakened cotton resistance to Verticillium wilt.
- Taxon
- Gossypium hirsutumGossypium hirsutumt0
- Treatment
- silencing GHHIT4_4e0
- Interactor
- VerticilliumVerticilliume0pathogen
- Gene
- GHHIT4_4e0
- Outcome
- resistance to Verticillium wilte0
- Direction
- weakenede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“silencing GHHIT4_4 had a significant impact, the resistance to Verticillium wilt weakened, and the internode length of the plants significantly decreased by 30.7% while the number of true leaves increased by 41.5%.”
t0“Gossypium hirsutum”
Claim · model-written summary
Silencing GHHIT4_4 decreased cotton internode length by 30.7%.
- Taxon
- Gossypium hirsutumGossypium hirsutumt0
- Treatment
- silencing GHHIT4_4e0
- Gene
- GHHIT4_4e0
- Outcome
- internode lengthe0
- Direction
- decreasede0
- Value
- 30.7%e0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“silencing GHHIT4_4 had a significant impact, the resistance to Verticillium wilt weakened, and the internode length of the plants significantly decreased by 30.7% while the number of true leaves increased by 41.5%.”
t0“Gossypium hirsutum”
Claim · model-written summary
Silencing GHHIT4_4 increased true leaf number by 41.5%.
- Taxon
- Gossypium hirsutumGossypium hirsutumt0
- Treatment
- silencing GHHIT4_4e0
- Gene
- GHHIT4_4e0
- Outcome
- number of true leavese0
- Direction
- increasede0
- Value
- 41.5%e0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“silencing GHHIT4_4 had a significant impact, the resistance to Verticillium wilt weakened, and the internode length of the plants significantly decreased by 30.7% while the number of true leaves increased by 41.5%.”
t0“Gossypium hirsutum”
Claim · model-written summary
GHHIT4_4 indirectly regulated PAL, 4CL, and CHI to enhance cotton resistance.
- Taxon
- cottonGossypium hirsutume0
- Interactor
- VerticilliumVerticilliume0pathogen
- Gene
- GHHIT4_4e0
- Outcome
- resistance to Verticillium wilte0
- Qualifier
- by indirectly regulating the PAL, 4CL, and CHI genese0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“GHHIT4_4 mainly enhanced cotton resistance to Verticillium wilt by indirectly regulating the PAL, 4CL, and CHI genes.”
Claim · model-written summary
GHHIT4_4 localized predominantly in mitochondria and nucleus.
- Taxon
- Gossypium hirsutumGossypium hirsutumt0
- Gene
- GHHIT4_4e0
- Outcome
- distributed in the mitochondria and nucleuse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“GHHIT4_4 was predominantly distributed in the mitochondria and nucleus.”
t0“Gossypium hirsutum”
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