Wounded plant cells can form callus to seal the wound site. Alternatively, wounding can cause adventitious organogenesis or somatic embryogenesis. These distinct developmental pathways require specific cell fate decisions. Here, we identify GhTCE1, a basic helix–loop–helix family transcription factor, and its interacting partners as a central regulatory module of early cell fate transition during in vitro dedifferentiation of cotton (Gossypium hirsutum). RNAi- or CRISPR/Cas9-mediated loss of GhTCE1 function resulted in excessive accumulation of reactive oxygen species (ROS), arrested callus cell elongation, and increased adventitious organogenesis. In contrast, GhTCE1-overexpressing tissues underwent callus cell growth, but organogenesis was repressed. Transcriptome analysis revealed that several pathways depend on proper regulation of GhTCE1 expression, including lipid transfer pathway components, ROS homeostasis, and cell expansion. GhTCE1 bound to the promoters of the target genes GhLTP2 and GhLTP3, activating their expression synergistically, and the heterodimer TCE1-TCEE1 enhances this activity. GhLTP2- and GhLTP3-deficient tissues accumulated ROS and had arrested callus cell elongation, which was restored by ROS scavengers. These results reveal a unique regulatory network involving ROS and lipid transfer proteins, which act as potential ROS scavengers. This network acts as a switch between unorganized callus growth and organized development during in vitro dedifferentiation of cotton cells.

The potassium transporter family HAK/KUP/KT is a large group of proteins that are important in plant potassium transport and play a crucial role in plant growth and development. The members of the family play an important role in the response of plants to abiotic stress by maintaining osmotic balance. However, the function of the family in cotton is unclear. In this study, whole genome identification and characterization of the HAK/KUP/KT family from upland cotton (Gossypium hirsutum) were carried out. Bioinformatics methods were used to identify HAK/KUP/KT family members from the G. hirsutum genome and to analyse the physical and chemical properties, basic characteristics, phylogeny, chromosome location and expression of HAK/KUP/KT family members. A total of 41 HAK/KUP/KT family members were identified in the G. hirsutum genome. Phylogenetic analysis grouped these genes into four clusters (I, II, III, IV), containing 6, 10, 3 and 22 genes, respectively. Chromosomal distribution, gene structure and conserved motif analyses of the 41 GhHAK genes were subsequently performed. The RNA-seq data and qRT-PCR results showed that the family had a wide range of tissue expression patterns, and they responded to certain drought stresses. Through expression analysis, seven HAK/KUP/KT genes involved in drought stress were screened, and four genes with obvious phenotypes under drought stress were obtained by VIGS verification, which laid a theoretical foundation for the function of the cotton HAK/KUP/KT family.

F-box/LR (FBXL), Leucine-rich repeats in F-box proteins, belongs to the Skp1-Cullin1-F-box protein (SCF) E3 ligase family. FBXL genes play important roles in plant growth, such as plant hormones, responses to environmental stress, and floral organ development. Here, a total of 518 FBXL genes were identified and analyzed in six plant species. Phylogenetic analysis showed that AtFBXLs, VvFBXLs, and GrFBXLs were clustered into three subfamilies (Ⅰ-Ⅲ). Based on the composition of the F-box domain and carboxyl-terminal amino acid sequence, FBXL proteins were classified into three types (Type-A/-B/-C). Whole-genome duplication (WGD) along with tandem duplications and segmental contributed to the expansion of this gene family. The result indicates that four cotton species are also divided into three subfamilies. FBXLs in cotton were classified into three clades by phylogenetic and structural analyses. Furthermore, expression analyses indicated that the expression patterns of GhFBXLs in different cotton tissues were different. The highly expressed of GH_A07G2363 in 5–8 mm anthers, indicates that this gene might play a role in the reproductive process, providing candidate genes for future studies on cotton fertility materials. This study provides an original functional opinion and a useful interpretation of the FBXL protein family in cotton.

Objective: Yield traits are crucial for cotton breeding. Analyzing the yield traits of terrestrial cotton and exploring their genetic mechanisms through a primary gene + multigene hybrid genetic model provide a theoretical basis for selecting high-quality cotton varieties and identifying associated molecular markers. Methods: Completing the construction of the six populations (P1, P2, F1, F2, B1, B2) using Xinluzhong 37 as the female parent and Xinluzhong 51 as the male parent. Six yield traits were assessed: single boll weight, boll number per plant, lint yield per plant, seed cotton per plant, lint percentage, and seed index. Data were tested for normal distribution, and the inheritance patterns of yield traits were analyzed through combined primary gene + polygenic analysis. Results: The coefficients of variation for the six yield traits ranged from 37.368% to 53.905%, 33.335% to 58.524%, 34.132% to 57.686%, 8.721% to 12.808%, 1.842% to 6.283%, and 8.783% to 12.580%, respectively. These traits displayed either normal or skewed normal distributions. The optimal genetic model for single boll weight and seed index was PG-ADI, while MX2-ADI-AD best fit the traits of boll number per plant and lint percentage. For lint yield per plant and seed cotton per plant, the 2MG-ADI model was optimal. The polygenic heritability for single boll weight was 29.58%; for boll number per plant, main gene heritability was 25.19%, with 0% heritability for polygenes; for lint yield per plant, the heritability of the main gene was 23.47%. For seed cotton per plant, the heritability of main genes was 15.38%, with lint percentage showing 63.25% heritability for main genes and 0.08% for polygenes, and seed index with 45.93% heritability due to polygenes. Overall, single boll weight and seed index were predominantly polygenic, while boll number per plant and lint percentage were largely controlled by main gene inheritance. The inheritance of lint yield per plant and seed cotton per plant was also primarily governed by main genes.