Flowering, the predominant reproductive strategy in the plant life cycle, is an intricate process regulated by both genetic and environmental factors. In Gossypium hirsutum, flowering time significantly influences adaptation and yield. Here, we identify the transcription factor GhCIL1 as a pivotal regulator promoting flowering in this crucial crop. Overexpression of GhCIL1 presented significantly shorter bolting and flowering time in Arabidopsis thaliana, while the silencing of GhCIL1 in cotton has been shown to delay bud appearance and flowering. Through the joint analysis of transcriptome based on the OE-GhCIL1 and CR-GhCIL1, it was found that GhCIL1 may regulate GhSWEET15-D04, GhSWEET15-A04 and GhPRR5. Electrophoretic mobility shift assays (EMSAs) and Dual-luciferase assays demonstrated that GhCIL1 could directly bind to and promote the expression of GhSWEET15-D04, GhSWEET15-A04, GhPRR5, and GhFT. The GhCIL1 possesses the capacity to form homodimers independently or heterodimers in conjunction with GhPRE6. silencing of GhPRE6 in cotton has been shown to promote bud appearance and flowering. Dual-luciferase assays and EMSAs demonstrated that the cotransformation of GhCIL1 and GhPRE6 inhibited the promotion and binding effect of GhCIL1 on GhSWEET15-D04, GhSWEET15-A04, GhPRR5, and GhFT. In conclusion, our work unveils a novel the flowering regulation pathway in cotton.

Aquaporins (AQPs) facilitate transmembrane water transport and play a crucial role in plant adaptation to diverse abiotic stress conditions. Plasma membrane Intrinsic Proteins (PIPs) are a crucial subgroup of aquaporins that facilitate water transport across plant cell membrane. However, the role of PIPs under salt stress responses in cotton remains partially unexplored. In this study, we found that the expression level of the aquaporin gene GhPIP2;1 was significantly upregulated under salt stress at three-leaf stage in upland cotton. The overexpression of GhPIP2;1 in Arabidopsis thaliana and virus-induced gene silencing (VIGS) in upland cotton evidenced that GhPIP2;1 positively regulate salt tolerance function. Biochemical analysis including superoxide dismutase (SOD), peroxidase (POD) activities and malondialdehyde (MDA) content measurements indicated that GhPIP2;1 could enhance stress tolerance by regulating the reactive oxygen species (ROS). The Electrophoretic mobility shift assay (EMSA) and dual luciferase reporter assay confirmed that GhNAC072 specifically binds to the promoter region of GhPIP2;1 and enhance the expression of GhPIP2;1. Phenotypic observations and physiological measurements of GhNAC072 overexpressed Arabidopsis and silenced cotton plants demonstrated that GhNAC072 positively responds to salt stress. The expression level of GhPIP2;1 was significantly decreased in GhNAC072-silenced plants, indicating that GhNAC072 enhances salt tolerance by regulating GhPIP2;1 expression. In summary, this study systematically confirmed that GhPIP2;1 may involve in salt tolerance mechanism in cotton and its expression is regulated by GhNAC072. These results will provide gene resources for salt tolerance and enrich the knowledge on the mechanism of salt tolerance in cotton.

Upland cotton (Gossypium hirsutum) is a principal economic crop and a fundamental raw material for the textile industry. The quality of cotton fibres is significantly influenced by the synthesis of cell wall polysaccharides. This study focuses on GhIRX10, a beta-1,4-xylosyltransferase crucial for xylan backbone synthesis. Overexpression of GhIRX10 enhances xylan synthesis, which impacts fibre elongation and secondary cell wall thickening. GhMYB102, identified as a direct regulator of GhIRX10 expression, was confirmed through comprehensive validation. Overexpression of GhMYB102 resulted in a similar phenotype as OE-GhIRX10: increased cell wall thickness and reduced fibre length. Overexpression of GhMYB102 upregulated the expression of key cell wall synthesis-related genes, including GhCESA4/7/8, GhIRXs, GhCESAs, GhGUXs, GhTBLs, GhXTHs, and GhXXTs. Consequently, the cellulose and hemicellulose contents in OE-GhMYB102 lines were significantly increased. GhMYB102 was also validated as a target gene regulated by GhFSN1 and GhMYB7, with the ability to reciprocally regulate GhFSN1 expression. In summary, we propose a regulatory model where GhMYB102 promotes the expression of GhIRX10 and other cell wall-related genes, thereby affecting fibre quality. This study elucidates the regulatory network of secondary cell wall synthesis in cotton and provides potential targets for improving fibre quality through molecular breeding.

Flowering time, a core determinant of early-maturing in cotton, is regulated by an intricate network involving photoperiod, autonomous, vernalization, gibberellin, and temperature signaling pathways, with FLOWERING LOCUS T (FT) serving as a central floral integrator. GhFT, a homolog gene of FT, could accelerate flowering time in cotton, while the regulatory mechanisms of GhFT remain incompletely characterized. This study identified GhAUX22D as a novel GhFT-interacting protein through yeast two-hybrid (Y2H) screening, with interactions validated by LCA (Luciferase Complementation Assay) and BiFC (Bimolecular Fluorescence Complementation) assays. Spatiotemporal expression profiling and in situ hybridization revealed GhAUX22D's involvement in cotton maturation processes. Functional validation using VIGS-suppressed cotton and transgenic Arabidopsis demonstrated GhAUX22D's positive role in flowering time. Crucially, auxin signaling was found to enhance GhFT-GhAUX22D interaction, suggesting a phytohormonal regulatory layer in flowering control. This study advances our understanding of flowering time mechanisms in cotton and contributes to the cultivation and breeding of early-maturing cotton cultivars.

Cotton is regarded as one of the significant economic crops in China, and its earliness is defined as one of the crucial traits influencing fiber quality and yield. To study the physiological and biochemical mechanisms related to early-maturing traits of cotton, cotton shoot apexes at the one-leaf, three-leaf, and five-leaf stages of the early-maturing cotton CCRI50 and late-maturing cotton Guoxinmian11 were collected for transcriptome sequencing and metabolomics, respectively. A total of 616, 782, and 842 differentially expressed genes (DEGs) at the one-leaf stage, three-leaf stage, and five-leaf stage were obtained through transcriptome sequencing, respectively. The metabolic detection results showed that 68, 56, and 62 differential metabolites (DMs) were obtained in the three periods, respectively. A total of 10 DMs were detected simultaneously from the one-leaf to five-leaf stage, 4 of which were phenolic acids and down-regulated in the early maturing variety CCRI50. A combined transcriptomic and metabolomic analysis revealed that phenylpropanoid biosynthesis, tyrosine metabolism, and phenylalanine metabolism might be important metabolic pathways in cotton bud differentiation. GhTYDC-A01 was identified in both the tyrosine metabolism and phenylalanine metabolism pathways, and it was highly expressed in pistils. To investigate the function of this gene in flowering, we overexpressed it in Arabidopsis thaliana. Compared to the wild type, the flowering time of the overexpression of GhTYDC-A01 in Arabidopsis was delayed. This study provides valuable resources and new insights into the relationship between metabolites and early-maturing cotton.

Flowering represents a pivotal developmental transition stage in the life cycle of a plant, and the occurrence of flowering at the optimal time is critical for reproductive success. WRKY transcription factors play a vital role in a signaling network that governs a multitude of plant processes. Here, a gene, GhWRKY46, that was differentially expressed in early and late maturing materials was identified via association analysis, and it was specifically expressed in flower buds. Under natural light and temperature conditions, compared to Jin668, the flowering time of OE-GhWRKY46 plants was advanced by approximately 6 days, while the flowering time of CR-GhWRKY46 plants was delayed by approximately 8 days. Transcriptomic data indicated that overexpression or knockout of GhWRKY46 resulted in the activation or repression, respectively, of the photoperiod gene CO-Like and genes related to bud differentiation. Combined RNA-seq and DAP-seq analysis revealed that three genes, namely, GhCOL4, GhCOL2 and GhFPF1-like, may be expressed downstream of GhWRKY46. Dual-luciferase assays and electrophoretic mobility shift assays (EMSAs) demonstrated that GhWRKY46 could directly bind to the W-box and promote the expression of these genes. Similarly, GhFT was also found to be activated by GhWRKY46. Both in vivo and in vitro biochemical analyses demonstrated that GhWRKY46 interacted with GhGAI, and GhGAI could interfere with the transcriptional activation of GhWRKY46, which in turn inhibited the expression of GhCOL4, GhCOL2, GhFPF1-like, and GhFT. In conclusion, this study accurately predicted the GhWRKY46 binding motif, which is important for the construction of regulatory networks of the WRKY family in other crops and introduces a novel regulatory module for the flowering regulation pathway in cotton.

Xyloglucan endotransglucosylase/hydrolases (XTHs) are cell wall modifying enzymes that function in response to multiple abiotic stresses by promoting cell wall reinforcement in plants. However, it is elusive whether GhXTH22 plays a role in cold stress tolerance in upland cotton (Gossypium hirsutum L.). In this study, we found that Arabidopsis (Arabidopsis thaliana (L.) Heynh) transgenic lines overexpressing GhXTH22 exhibit clearly increased cold tolerance compared with wild type (WT). Moreover, cold stress results in markable enhancements in the activities of catalase (CAT) and peroxidase (POD), cell wall rigidity, and the expression of cold responsive genes, as well as a notable reduction in malondialdehyde (MDA) levels in the overexpression (OE) lines compared with those in WT plants. Consistently, silencing GhXTH22 by virus-induced gene silencing (VIGS) method causes markedly decreases in cold stress tolerance in upland cotton. Additionally, by electrophoretic mobility shift assay (EMSA) and dual-luciferase assay, it was confirmed that GhMYB44 activate the expression of GhXTH22 by binding to AACNG motif on the promoter, and silencing the expression of GhMYB44 can decrease the cold resistance of cotton. In summary, this study systematically confirmed the cold resistance function of GhXTH22 and discovered the upstream regulating gene of GhXTH22, providing gene resources for cold resistance and enriching the knowledge on the mechanism of cold resistance.

BACKGROUND:Cotton fiber length and strength are both key traits of fiber quality, and fiber strength (FS) is tightly correlated with secondary cell wall (SCW) biosynthesis. The three-amino-acid-loop-extension (TALE) superclass homeoproteins are involved in regulating diverse biological processes in plants, and some TALE members has been identified to play a key role in regulating SCW formation. However, little is known about the functions of TALE members in cotton (Gossypium spp.). RESULTS:In the present study, based on gene homology, 46, 47, 88 and 94 TALE superfamily genes were identified in G. arboreum, G. raimondii, G. barbadense and G. hirsutum, respectively. Phylogenetic and evolutionary analysis showed the evolutionary conservation of two cotton TALE families (including BEL1-like and KNOX families). Gene structure analysis also indicated the conservation of GhTALE members under selection. The analysis of promoter cis-elements and expression patterns suggested potential transcriptional regulation functions in fiber SCW biosynthesis and responses to some phytohormones for GhTALE proteins. Genome-wide analysis of colocalization of TALE transcription factors with SCW-related QTLs revealed that some BEL1-like genes and KNAT7 homologs may participate in the regulation of cotton fiber strength formation. Overexpression of GhKNAT7-A03 and GhBLH6-A13 significantly inhibited the synthesis of lignocellulose in interfascicular fibers of Arabidopsis. Yeast two-hybrid (Y2H) experiments showed extensive heteromeric interactions between GhKNAT7 homologs and some GhBEL1-like proteins. Yeast one-hybrid (Y1H) experiments identified the upstream GhMYB46 binding sites in the promoter region of GhTALE members and defined the downstream genes that can be directly bound and regulated by GhTALE heterodimers. CONCLUSION:We comprehensively identified TALE superfamily genes in cotton. Some GhTALE members are predominantly expressed during the cotton fiber SCW thicking stage, and may genetically correlated with the formation of FS. Class II KNOX member GhKNAT7 can interact with some GhBEL1-like members to form the heterodimers to regulate the downstream targets, and this regulatory relationship is partially conserved with Arabidopsis. In summary, this study provides important clues for further elucidating the functions of TALE genes in regulating cotton growth and development, especially in the fiber SCW biosynthesis network, and it also contributes genetic resources to the improvement of cotton fiber quality.

Climate change severely affects crop production. Cotton is one of the primary fiber crops in the world and its production is susceptible to various environmental stresses, especially drought and salinity. Development of stress tolerant genotypes is the only way to escape from these environmental constraints. We identified sixteen homologs of the Arabidopsis JUB1 gene in cotton. Expression of GhJUB1_3-At was significantly induced in the temporal expression analysis of GhJUB1 genes in the roots of drought tolerant (H177) and susceptible (S9612) cotton genotypes under drought. The silencing of the GhJUB1_3-At gene alone and together with its paralogue GhJUB1_3-Dt reduced the drought tolerance in cotton plants. The transgenic lines exhibited tolerance to the drought and salt stress as compared to the wildtype (WT). The chlorophyll and relative water contents of wildtype decreased under drought as compared to the transgenic lines. The transgenic lines showed decreased H2O2 and increased proline levels under drought and salt stress, as compared to the WT, indicating that the transgenic lines have drought and salt stress tolerance. The expression analysis of the transgenic lines and WT revealed that GAI was upregulated in the transgenic lines in normal conditions as compared to the WT. Under drought and salt treatment, RAB18 and RD29A were strongly upregulated in the transgenic lines as compared to the WT. Conclusively, GhJUB1_3-At is not an auto activator and it is regulated by the crosstalk of GhHB7, GhRAP2-3 and GhRAV1. GhRAV1, a negative regulator of abiotic stress tolerance and positive regulator of leaf senescence, suppresses the expression of GhJUB1_3-At under severe circumstances leading to plant death.

Cotton architecture is determined by the differentiation fate transition of axillary meristem (AM), and influences cotton yield and the efficiency of mechanized harvesting. We observed that the initiation of flowering primordium was earlier in early-maturing than that in late-maturing cultivars during the differentiation and development of AM. The RNA-Seq and expression level analyses showed that genes FLAVIN BINDING, KELCH REPEAT, F-BOX1 (GhFKF1), and GIGANTEA (GhGI) were in response to circadian rhythms, and involved in the regulation of cotton flowering. The gene structure, predicted protein structure, and motif content analyses showed that in Arabidopsis, cotton, rapseed, and soybean, proteins GhFKF1 and GhGI were functionally conserved and share evolutionary origins. Compared to the wild type, in GhFKF1 mutants that were created by the CRISPR/Cas9 system, the initiation of branch primordium was inhibited. Conversely, the knocking out of GhGI increased the number of AM differentiating into flower primordium, and there were much more lateral branch differentiation and development. Besides, we investigated that proteins GhFKF1 and GhGI can interact with each other. These results suggest that GhFKF1 and GhGI are key regulators of cotton architecture development, and may collaborate to regulate the differentiation fate transition of AM, ultimately influencing plant architecture. We describe a strategy for using the CRISPR/Cas9 system to increase cotton adaptation and productivity by optimizing plant architecture.

Multiple abiotic stresses, such as drought and high salinity, seriously limit cotton production. However, the mitogen-activated protein kinases (MAPKs) could play a critical role in responding to multiple environmental abiotic stresses. In this study, one of the MAPKs in cotton, GhMAPK3 (GH_D03G1517), was cloned and its role in drought and salt stress tolerance was investigated. The qRT-PCR assays demonstrated that GhMAPK3 predominantly expressed in leaf and root parts and showed higher expression under salt and drought stress. Subcellular localization assays revealed that GhMAPK3 was distributed in the nucleus and cytoplasm. The silenced of GhMAPK3 could decrease the tolerance of cotton plants under salt and drought stress via virus-induced gene silencing (VIGS) experiments. In GhMAPK3-slienced plants, the expression of stress-responsive genes has been revealed to be down-regulated levels, while the oxidants and antioxidant enzymes showed relatively higher levels and relatively lower levels, respectively. We further acquired three GhMAPK3 overexpression Arabidopsis lines, and they showed higher drought and salt tolerance than the wild-type (WT) plants, the chlorophyll content and relative leaf water content were found to be relatively stable through GhMAPK3 overexpressed plants under the abiotic stress. Further investigation revealed that GhMAPK3 showed a higher germination rate and longer root length under salt and drought stress. Overall, our findings revealed that the GhMAPK3 gene plays a prominent role in drought and salt tolerance and offers knowledge on crop genetic improvements.

Vascular Plant One‑zinc Finger (VOZ) transcription factor can respond to a variety of abiotic stresses, however its function in cotton and the molecular mechanisms of response to salt tolerance remained unclear. In this study, we found that GhVOZ1 is highly expressed in stamen and stem of cotton under normal conditions. The expression of GhVOZ1 increased significantly after 3 h of salt treatment in three-leaf staged upland cotton. Overexpressed transgenic lines of GhVOZ1 in Arabidopsis and upland cotton were treated with salt stress and we found that GhVOZ1 could respond positively to salt stress. GhVOZ1 can regulate Arabidopsis Vacuolar Proton Pump Pyrophosphatase (H+-PPase) gene (AVP1) expression through specific binding to GCGTCTAAAGTACGC site on GhAVP1 promoter, which was examined through Dual-luciferase assay and Electrophoretic mobility shift assay (EMSA). AVP1 expression was significantly increased in Arabidopsis with GhVOZ1 overexpression, while GhAVP1 expression was decreased in virus induced gene silenced (VIGS) cotton plants of GhVOZ1. Knockdown of GhAVP1 expression in cotton plants by VIGS showed decreased superoxide dismutase (SOD) and peroxidase (POD) activities, whereas an increased malondialdehyde (MDA) content and ultimately decreased salt tolerance. The GhVOZ1-AVP1 module could maintain sodium ion homeostasis through cell ion transport and positively regulate the salt tolerance in cotton, providing new ideas and insights for the study of salt tolerance.

Xyloglucan, an important hemicellulose, plays a crucial role in maintaining cell wall structure and cell elongation. However, the effects of xyloglucan on cotton fiber development are not well understood. GhMUR3 encodes a xyloglucan galactosyltransferase that is essential for xyloglucan synthesis and is highly expressed during fiber elongation. In this study, we report that GhMUR3 participates in cotton fiber development under the regulation of GhMYB30. Overexpression GhMUR3 affects the fiber elongation and cell wall thickening. Transcriptome showed that the expression of genes involved in secondary cell wall synthesis was prematurely activated in OE-MUR3 lines. In addition, GhMYB30 was identified as a key regulator of GhMUR3 by Y1H, Dual-Luc, and electrophoretic mobility shift assay (EMSA) assays. GhMYB30 directly bound the GhMUR3 promoter and activated GhMUR3 expression. Furthermore, DAP-seq of GhMYB30 was performed to identify its target genes in the whole genome. The results showed that many target genes were associated with fiber development, including cell wall synthesis-related genes, BR-related genes, reactive oxygen species pathway genes, and VLCFA synthesis genes. It was demonstrated that GhMYB30 may regulate fiber development through multiple pathways. Additionally, GhMYB46 was confirmed to be a target gene of GhMYB30 by EMSA, and GhMYB46 was significantly increased in GhMYB30-silenced lines, indicating that GhMYB30 inhibited GhMYB46 expression. Overall, these results revealed that GhMUR3 under the regulation of GhMYB30 and plays an essential role in cotton fiber elongation and secondary wall thickening. Additionally, GhMYB30 plays an important role in the regulation of fiber development and regulates fiber secondary wall synthesis by inhibiting the expression of GhMYB46.

AP1 is a MADS-Box gene that plays an important role in the flowering pathway of plants. Although GhAP1 has been widely studied, little is known about the proteins that interact with it in cotton. In this study, GhAP1 was used as a bait protein to screen a cDNA library for interacting proteins related to flowering. The results of yeast two-hybrid, bimolecular fluorescence complementation (BiFC) and coimmunoprecipitation (Co-IP) assays showed that GhGAI interacted with GhAP1. The qRTPCR results showed that GhGAI was highly expressed in the flower organs and leaves of upland cotton, and the expression of GhGAI between the early-maturing cultivar and the late-maturing cultivar at the first, second, and fifth leaf stages was significantly different in the shoot apexes. Subcellular localization showed that GhGAI was located in the nucleus. Overexpression of GhGAI in Arabidopsis thaliana could lead to early flowering, increase the contents of GA3 and GA4 hormones, and inhibit the expression of the DELLA proteins AtRGL1 and AtRGL2, while promoting the expression of the flowering core factors AtSOC1, AtLYF, AtFT, and AtAP1. Virus-induced gene silencing (VIGS) of GhGAI in cotton delayed flowering. In silenced plants, the expression of GhRGL1 and GhRGL2 was increased, and the expression of GhSOC1, GhLYF, GhFT, and GhAP1 was decreased. Our results provide important genetic resources for the molecular breeding of early-maturing varieties and lay a foundation for exploring the flowering regulatory network in cotton.

The CCCH zinc-finger protein contains a typical C3H-type motif widely existing in plants, and it plays an important role in plant growth, development, and stress responses. In this study, a CCCH zinc-finger gene, GhC3H20, was isolated and thoroughly characterized to regulate salt stress in cotton and Arabidopsis. The expression of GhC3H20 was up-regulated under salt, drought, and ABA treatments. GUS activity was detected in the root, stem, leaves, and flowers of ProGhC3H20::GUS transgenic Arabidopsis. Compared with the control, the GUS activity of ProGhC3H20::GUS transgenic Arabidopsis seedlings under NaCl treatment was stronger. Through the genetic transformation of Arabidopsis, three transgenic lines of 35S-GhC3H20 were obtained. Under NaCl and mannitol treatments, the roots of the transgenic lines were significantly longer than those of the wild-type (WT) Arabidopsis. The leaves of the WT turned yellow and wilted under high-concentration salt treatment at the seedling stage, while the leaves of the transgenic Arabidopsis lines did not. Further investigation showed that compared with the WT, the content of catalase (CAT) in the leaves of the transgenic lines was significantly higher. Therefore, compared with the WT, overexpression of GhC3H20 enhanced the salt stress tolerance of transgenic Arabidopsis. A virus-induced gene silencing (VIGS) experiment showed that compared with the control, the leaves of pYL156-GhC3H20 plants were wilted and dehydrated. The content of chlorophyll in pYL156-GhC3H20 leaves was significantly lower than those of the control. Therefore, silencing of GhC3H20 reduced salt stress tolerance in cotton. Two interacting proteins (GhPP2CA and GhHAB1) of GhC3H20 have been identified through a yeast two-hybrid assay. The expression levels of PP2CA and HAB1 in transgenic Arabidopsis were higher than those in the WT, and pYL156-GhC3H20 had expression levels lower than those in the control. GhPP2CA and GhHAB1 are the key genes involved in the ABA signaling pathway. Taken together, our findings demonstrate that GhC3H20 may interact with GhPP2CA and GhHAB1 to participate in the ABA signaling pathway to enhance salt stress tolerance in cotton.

Additional file 1: Table S1. Basic information and biophysical properties of the predicted DUF4228 proteins in cotton.

BACKGROUND:Phloem protein 2 (PP2) proteins play a vital role in the Phloem-based defense (PBD) and participate in many abiotic and biotic stress. However, research on PP2 proteins in cotton is still lacking.RESULTS:A total of 25, 23, 43, and 47 PP2 genes were comprehensively identified and characterized in G.arboretum, G.raimondii, G.barbadense, and G.hirsutum. The whole genome duplication (WGD) and allopolyploidization events play essential roles in the expansion of PP2 genes. The promoter regions of GhPP2 genes contain many cis-acting elements related to abiotic stress and the weighted gene co-expression network analysis (WGCNA) analysis displayed that GhPP2s could be related to salt stress. The qRT-PCR assays further confirmed that GhPP2-33 could be dramatically upregulated during the salt treatment. And the virus-induced gene silencing (VIGS) experiment proved that the silencing of GhPP2-33 could decrease salt tolerance.CONCLUSIONS:The results in this study not only offer new perspectives for understanding the evolution of PP2 genes in cotton but also further explore their function under salt stress.

Background Cotton, being extensively cultivated, holds immense economic significance as one of the most prominent crops globally. The SET (Su(var), E, and Trithorax) domain-containing protein is of significant importance in plant development, growth, and response to abiotic stress by modifying the lysine methylation status of histone. However, the comprehensive identification of SET domain genes (SDG) have not been conducted in upland cotton ( Gossypium hirsutum L.). Results A total of 229 SDGs were identified in four Gossypium species, including G. arboretum , G. raimondii , G. hirsutum , and G. barbadense . These genes could distinctly be divided into eight groups. The analysis of gene structure and protein motif revealed a high degree of conservation among the SDGs within the same group. Collinearity analysis suggested that the SDGs of Gossypium species and most of the other selected plants were mainly expanded by dispersed duplication events and whole genome duplication (WGD) events. The allopolyploidization event also has a significant impact on the expansion of SDGs in tetraploid Gossypium species. Furthermore, the characteristics of these genes have been relatively conserved during the evolution. Cis -element analysis revealed that GhSDGs play a role in resistance to abiotic stresses and growth development. Furthermore, the qRT-PCR results have indicated the ability of GhSDGs to respond to salt stress. Co-expression analysis revealed that GhSDG51 might co-express with genes associated with salt stress. In addition, the silencing of GhSDG51 in cotton by the virus-induced gene silencing (VIGS) method suggested a potential positive regulatory role of GhSDG51 in salt stress. Conclusions The results of this study comprehensively analyze the SDGs in cotton and provide a basis for understanding the biological role of SDGs in the stress resistance in upland cotton.

(1) Background: Mapping QTLs for early-maturing traits is necessary for the development of early-maturing variety breeding. (2) Methods: In this research, a high-density genetic map (HDGM) was constructed using an F2 population with 100 individuals and single nucleotide polymorphism markers (SNPs) developed using the genotyping-by-sequencing (GBS) method. (3) Results: The HDGM, which covered a total distance of 3167.14 cM, harbored 5454 SNPs with an average marker interval of 0.58 cM. In total, 18 QTLs for four early-maturing characters were detected and explained 11.6–46.4% of phenotypic variation (PV). Two QTLs of the whole growing period (WGP) and height of the node of the first fruiting branch (HNFFB) were identified as stable QTLs. In total, 125 candidate genes were identified in the confidence intervals of these stable QTLs. Presumably, Gh_D03G0857 may play an important role in regulating earliness. (4) Conclusions: This research will provide new information about fine mapping of QTLs for earliness traits, molecular marker assisted selection (MAS) of earliness traits, and pyramiding breeding as well.

Salt stress is a major abiotic stressor that can severely limit plant growth, distribution, and crop yield. DNA-binding with one finger (Dof) is a plant-specific transcription factor that plays a crucial role in plant growth, development, and stress response. In this study, the function of a Dof transcription factor, GhDof1.7, was investigated in upland cotton. The GhDof1.7 gene has a coding sequence length of 759 base pairs, encoding 252 amino acids, and is mainly expressed in roots, stems, leaves, and inflorescences. Salt and abscisic acid (ABA) treatments significantly induced the expression of GhDof1.7. The presence of GhDof1.7 in Arabidopsis may have resulted in potential improvements in salt tolerance, as suggested by a decrease in H2O2 content and an increase in catalase (CAT) and superoxide dismutase (SOD) activities. The GhDof1.7 protein was found to interact with GhCAR4 (C2-domain ABA-related 4), and the silencing of either GhDof1.7 or GhCAR4 resulted in reduced salt tolerance in cotton plants. These findings demonstrate that GhDof1.7 plays a crucial role in improving the salt tolerance of upland cotton and provide insight into the regulation of abiotic stress response by Dof transcription factors.