Cell wall-associated receptor kinases (WAKs/WAKLs), are a specialized class of plant receptor kinases essential for signaling during stress conditions. However, there has been no report on the involvement of WAKs/WAKLs in salt tolerance in cotton. In this study, we report the functional characterization of GhWAKL26, whose expression is induced by salt, with its levels increasing over time and with higher salt concentrations. In addition, the fusion protein of GhWAKL26 and GFP was localized to the plasma membrane. In transgenic Arabidopsis, the dry weight, fresh weight, and root length were significantly higher than those of wild-type plants, indicating enhanced salt tolerance. While in GhWAKL26-silenced cotton seedlings, H2O2, O-2(-), and MDA content were increased, and chlorophyll content was reduced under salt stress, displaying compromised salt tolerance. RNA-seq analysis revealed that the silencing of GhWAKL26 resulted in the down-regulation of expression levels of certain ion transport-related genes under salt stress, concurrently leading to an increased Na+/K+ ratio in cotton seedlings. Overall, our findings indicate that GhWAKL26 enhanced plant resistance to salt stress in cotton by regulating the balance of Na+ and K+ ions.

We characterized the WAK gene family in Gossypium barbadense and revealed the potential function of GbWAK5 in regulating salt tolerance by modulating ion homeostasis. Soil salinization is one of the main factors restricting cotton production. Although the role of the wall-associated kinases (WAKs) in plants has been extensively studied, its response to salt stress in sea-island cotton (Gossypium barbadense L.) has not been reported. Here, we conducted a whole-genome analysis of the WAK gene family in G. barbadense, identifying a total of 70 GbWAK genes, which were classified into five clades. Segmental and tandem duplication events have contributed to the expansion of the GbWAK gene family. A large number of cis-acting elements were predicted in the GbWAK promoter region. Through RNA sequencing, 37 GbWAKs that potentially play a role in cotton's response to salt stress were screened out, among which 10 genes with sustained up-regulated expression were confirmed by quantitative real-time PCR (qRT-PCR). GbWAK5, a member of Clade II, was significantly up-regulated following NaCl treatment and exhibited a typical WAK structure. Subcellular localization indicated that GbWAK5 is localized on the plasma membrane. Virus-induced gene silencing (VIGS) experiments revealed that the knockdown of GbWAK5 resulted in more severe dehydration and wilting in plants compared to the control under NaCl treatment. RNA-seq analysis revealed that several ion transport-related genes were down-regulated in TRV:GbWAK5 plants under salt stress, while TRV:GbWAK5 plants accumulated more Na+ and exhibited a higher Na+/K+ ratio compared to TRV:00 plants. These results offer a comprehensive analysis of the G. barbadense WAK gene family for the first time, and conclude that GbWAK5 is a promising gene for improving cotton's resistance to salt stress.

Verticillium wilt (VW), Fusarium wilt (FW) and Root-knot nematode (RKN) are the main diseases affecting cotton production. However, many reported quantitative trait loci (QTLs) for cotton resistance have not been used for agricultural practices because of inconsistencies in the cotton genetic background. The integration of existing cotton genetic resources can facilitate the dis-covery of important genomic regions and candidate genes involved in disease resistance. Here, an improved and comprehensive meta-QTL analysis was conducted on 487 disease resistant QTLs from 31 studies in the last two decades. A consensus linkage map with genetic overall length of 3006.59 cM containing 8650 markers was constructed. A total of 28 Meta-QTLs (MQTLs) were discovered, among which nine MQTLs were identified as related to resistance to multiple diseases. Candidate genes were predicted based on public transcriptome data and enriched in pathways related to disease resistance. This study used a method based on the integration of Meta-QTL, known genes and transcriptomics to reveal major genomic regions and putative candidate genes for resistance to multiple diseases, providing a new basis for marker-assisted selection of high disease resistance in cotton breeding.

Key message The study of the origin, evolution, and diversification of the wall-associated kinase gene family in plants facilitates their functional investigations in the future. Abstract Wall-associated kinases (WAKs) make up one subfamily of receptor-like kinases (RLKs), and function directly in plant cell elongation and responses to biotic and abiotic stresses. The biological functions of WAKs have been extensively characterized in angiosperms; however, the origin and evolutionary history of the WAK family in green plants remain unclear. Here, we performed a comprehensive analysis of the WAK family to reveal its origin, evolution, and diversification in green plants. In total, 1061 WAK genes were identified in 37 species from unicellular algae to multicellular plants, and the results showed that WAK genes probably originated before bryophyte differentiation and were widely distributed in land plants, especially angiosperms. The phylogeny indicated that the land plant WAKs gave rise to five clades and underwent lineage-specific expansion after species differentiation. Cis -acting elements and expression patterns analyses of WAK genes in Arabidopsis and rice demonstrated the functional diversity of WAK genes in these two species. Many gene gains and losses have occurred in angiosperms, leading to an increase in the number of gene copies. The evolutionary trajectory of the WAK family during polyploidization was uncovered using Gossypium species. Our results provide insights into the evolution of WAK genes in green plants, facilitating their functional investigations in the future.

Histone demethylases containing JumonjiC (JmjC) domains regulate gene transcription and chromatin structure by changing the methylation status of lysine residues and play an important role in plant growth and development. In this study, a total of 332 JmjC family genes were identified from 21 different plant species. The evolutionary analysis results showed that the JmjC gene was detected in each species, that is, the gene has already appeared in algae. The phylogenetic analysis showed that the KDM3/JHDM2 subfamily genes may have appeared when plants transitioned from water to land, but were lost in lycophytes (Selaginella moellendorffii). During the evolutionary process, some subfamily genes may have been lost in individual species. According to the analysis of the conserved domains, all of the plant JmjC genes contained a typical JmjC domain, which was highly conserved during plant evolution. The analysis of cis-acting elements showed that the promoter region of the JmjC gene was rich in phytohormones and biotic and abiotic stress-related elements. The transcriptome data analysis and protein interaction analyses showed that JmjC genes play an important role in plant growth and development. The results clarified the evolutionary history of JmjC family genes in plants and lay the foundation for the analysis of the biological functions of JmjC family genes.

The wall-associated kinases (WAKs) and WAK-like kinases (WAKLs) form a group of receptor-like kinases (RLKs) with extracellular domains tightly linked to the cell wall. The WAKs/WAKLs have been known to be involved in plant growth, development, and stress responses. However, the functions of WAKs/WAKLs are less well known in cotton. In this study, 58, 66, and 99 WAK/WAKL genes were identified in Gossypium arboreum, G. raimondii, and G. hirsutum, respectively. Phylogenetic analysis showed they were classified into five groups, with two groups specific to cotton. Collinearity analysis revealed that segmental and tandem duplications resulted in expansion of the WAK/WAKL gene family in cotton. Moreover, the Ka/Ks ratios indicated this family was exposed to purifying selection pressure during evolution. The structures of the GhWAK/WAKL genes and encoded proteins suggested the functions of WAKs/WAKLs in cotton were conserved. Transient expression of four WAK/WAKL-GFP fusion constructs in Arabidopsis protoplasts indicated that they were localized on the plasma membrane. The cis-elements in the GhWAK/WAKL promoters were responsive to multiple phytohormones and abiotic stresses. Expression profiling showed that GhWAK/WAKL genes were induced by various abiotic stresses. This study provides insights into the evolution of WAK/WAKL genes and presents fundamental information for further analysis in cotton.