Genome-Wide and Expression Pattern Analysis of the HIT4 Gene Family Uncovers the Involvement of GHHIT4_4 in Response to Verticillium Wilt in Gossypium hirsutum.

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.

Function analysis of GhWRKY53 regulating cotton resistance to verticillium wilt by JA and SA signaling pathways.

WRKY transcription factors (TFs) play an important role in regulating the mechanism of plant self-defense. However, the function of most WRKY TFs in upland cotton (Gossypium hirsutum) is still unknown. Hence, studying the molecular mechanism of WRKY TFs in the resistance of cotton to Verticillium dahliae is of great significance to enhancing cotton disease resistance and improving its fiber quality. In this study, Bioinformatics has been used to characterize the cotton WRKY53 gene family. we analyzed the GhWRKY53 expression patterns in different resistant upland cotton cultivars treated with salicylic acid (SA) and methyl jasmonate (MeJA). Additionally, GhWRKY53 was silenced using a virus-induced gene silencing (VIGS) to determine the contribution of GhWRKY53 to V. dahliae resistance in cotton. The result showed that GhWRKY53 mediated SA and MeJA signal transduction pathways. After VIGS of the GhWRKY53, the ability of cotton to resist V. dahliae decreased, indicating that the GhWRKY53 could be involved in the disease resistance mechanism of cotton. Studies on the levels of SA and jasmonic acid (JA) and their related pathway genes demonstrated that the silencing of GhWRKY53 inhibited the SA pathway and activated the JA pathway, thereby reducing the resistance of plants to V. dahliae. In conclusion, GhWRKY53 could change the tolerance of upland cotton to V. dahliae by regulating the expression of SA and JA pathway-related genes. However, the interaction mechanism between JA and SA signaling pathways in cotton in response to V. dahliae requires further study.

Influence of Large Steric Hinderance Substituent Position on Conformation and Charge Transfer Process for Non-Fused Ring Acceptors.

To obtain stable and planar molecular geometry in non-fused electron acceptors, A4T-25 and A4T-26 are designed and synthesized by introducing the bulk 2,4,6-triisopropylphenyl side groups onto different positions of the central two thiophene units. A4T-25 and A4T-26 both show a narrow-bandgap of 1.39 and 1.46 eV, with highest occupied molecular orbital/lowest unoccupied molecular orbital levels of -5.56/-3.81 and -5.65/-3.83 eV, respectively, and the electrostatic potential distributions imply that they have strong electron-accepting capability. The single crystal structure analysis and the transfer integral calculation demonstrate that the much more compact π-π stacking in A4T-26 can promote efficient charge transportation compared to that in A4T-25. Therefore, the electron mobility of A4T-26 is obviously higher and more balanced than that of A4T-25. When blending the two acceptors with the same polymer donor PBDB-TF, the photovoltaic cell based on PBDB-TF:A4T-25 has an inadequate power conversion efficiency (PCE) of 7.83%, while the PBDB-TF:A4T-26-based one yields an enhanced PCE of 12.1%. Overall, this study offers an insight into the relationship between the fine-tuning of the molecular structure of non-fused ring acceptors and the corresponding charge transfer process in organic solar cells.

Completely non-fused electron acceptor with 3D-interpenetrated crystalline structure enables efficient and stable organic solar cell.

Non-fullerene acceptors (NFAs) based on non-fused conjugated structures have more potential to realize low-cost organic photovoltaic (OPV) cells. However, their power conversion efficiencies (PCEs) are much lower than those of the fused-ring NFAs. Herein, a new bithiophene-based non-fused core (TT-Pi) featuring good planarity as well as large steric hindrance was designed, based on which a completely non-fused NFA, A4T-16, was developed. The single-crystal result of A4T-16 reveals that a three-dimensional interpenetrating network can be formed due to the compact π-π stacking between the adjacent end-capping groups. A high PCE of 15.2% is achieved based on PBDB-TF:A4T-16, which is the highest value for the cells based on the non-fused NFAs. Notably, the device retains ~84% of its initial PCE after 1300 h under the simulated AM 1.5 G illumination (100 mW cm-2). Overall, this work provides insight into molecule design of the non-fused NFAs from the aspect of molecular geometry control.