The soil in Yuncheng Salt Lake has serious salinization and the biogeographic environment affects the composition and distribution of special halophilic and salt-tolerant microbial communities in this area. Therefore, this study collected soils at distances of 15, 30, and 45 m from the Salt Lake and used non-saline soil (60 m) as a control to explore the microbial composition and salt tolerance mechanisms using metagenomics technology. The results showed that the dominant species and abundance of salt-tolerant microorganisms changed gradually with distance from Salt Lake. The salt-tolerant microorganisms can increase the expression of the Na⁺/H⁺ antiporter by upregulating the Na⁺/H⁺ antiporter subunit mnhA-G to respond to salt stress, simultaneously upregulating the genes in the betaine/proline transport system to promote the conversion of choline into betaine, while also upregulating the trehalose/maltose transport system encode genes to promote the synthesis of trehalose to resist a high salt environment.

Sea-island cotton (Gossypium barbadense) has drawn great attention in the textile industry for its comprehensive resistance and superior fiber properties. However, the mechanisms involved in fiber growth and development are unclear. As TCP transcription factors play important roles in plant growth and development, this study investigated the TCP family genes in G. barbadense (GbTCP). We identified 75 GbTCP genes, of which 68 had no introns. Phylogenetic analyses categorized the GbTCP transcription factors into 11 groups. Genomic analyses showed that 66 genes are located on 21 chromosomes. Phylogenetic analyses of G. arboreum, G. raimondii, G. hirsutum, G. barbadense, Theobroma cacao, Arabidopsis thaliana, Oryza sativa, Sorghum bicolor, and Zea mays, Picea abies, Sphagnum fallax and Physcomitrella patens, categorized 373 TCP genes into two classes (Classes I and II). By studying the structures of TCP genes in sea-island cotton, we identified genes from the same evolutionary branches that showed similar motif patterns. qRT-PCR results suggested that the GbTCPs had different expression patterns in fibers at various developmental stages of cotton, with several showing specific expression patterns during development. This report helps lay the foundation for future investigations of TCP functions and molecular mechanisms in sea-island cotton fiber development.

In plant cells, vacuolar H⁺-ATPases (V-ATPases) are responsible for deacidification of the cytosol and energisation of the secondary transport processes across the tonoplast. A number of V-ATPase subunit genes have been demonstrated to be involved in the regulation of the plant response to water deficit. However, there are no reports on the role of V-ATPase subunit A (VHA-A) in dehydration tolerance of cotton. In this study, cotton GhVHA-A gene was functionally characterized, especially with regard to its role in dehydration stress tolerance. Expression analysis showed that GhVHA-A was differentially expressed in various cotton organs and was induced by dehydration, low temperature, high salinity, and abscisic acid treatment in leaves. We also report that GhVHA-A improve dehydration tolerance in transgenic tobacco and cotton. Virus-induced gene silencing of GhVHA-A decreased the tolerance of cotton plantlets to dehydration stress. Silencing GhVHA-A decreased chlorophyll content and antioxidant enzyme activities and increased malondialdehyde (MDA) content in cotton under dehydration stress. However, transgenic tobacco expressing GhVHA-A exhibited enhanced dehydration resistance, resulting in reduced leaf water loss, higher average root length, and lower MDA levels under dehydration stress. Meanwhile, overexpression of GhVHA-A in tobacco conferred water deficit tolerance by enhancing osmotic adjustment (proline) and the activities of the antioxidant enzymes superoxide dismutase and peroxidase, thereby enhancing reactive oxygen species detoxification. These results suggest that GhVHA-A plays an important role in conferring resistance to dehydration stress. Our results have identified GhVHA-A as a candidate gene for improving dehydration tolerance in plants.

Cyclophilins (CYPs) are a member of the immunophilin superfamily (in addition to FKBPs and parvulins) and play a significant role in peptidyl-prolyl cis-trans isomerase (PPIase) activity. Previous studies have shown that CYPs have important functions in plants, but no genome-wide analysis of the cotton CYP gene family has been reported, and the specific biological function of this gene is still elusive. Based on the release of the cotton genome sequence, we identified 75, 78, 40 and 38 CYP gene sequences from G. barbadense, G. hirsutum, G. arboreum, and G. raimondii, respectively; 221 CYP genes were unequally located on chromosomes. Phylogenetic analysis showed that 231 CYP genes clustered into three major groups and eight subgroups. Collinearity analysis showed that segmental duplications played a significant role in the expansion of CYP members in cotton. There were light-responsiveness, abiotic-stress and hormone-response elements upstream of most of the CYPs. In addition, the motif composition analysis revealed that 49 cyclophilin proteins had extra domains, including TPR (tetratricopeptide repeat), coiled coil, U-box, RRM (RNA recognition motif), WD40 (RNA recognition motif) and zinc finger domains, along with the cyclophilin-like domain (CLD). The expression patterns based on qRT-PCR showed that six CYP expression levels showed greater differences between Xinhai21 (long fibres, G. barbadense) and Ashmon (short fibres, G. barbadense) at 10 and 20 days postanthesis (DPA). These results signified that CYP genes are involved in the elongation stage of cotton fibre development. This study provides a valuable resource for further investigations of CYP gene functions and molecular mechanisms in cotton.

Verticillium wilt, caused by the ascomycete fungus Verticillium dahliae (Vd), is a devastating disease of numerous plant species. However, the pathogenicity/virulence-related genes in this fungus, which may be potential targets for improving plant resistance, remain poorly elucidated. For the study of these genes in Vd, we used a well-established host-induced gene silencing (HIGS) approach and identified 16 candidate genes, including a putative adenylate kinase gene (VdAK). Transiently VdAK-silenced plants developed milder wilt symptoms than control plants did. VdAK-knockout mutants were more sensitive to abiotic stresses and had reduced germination and virulence on host plants. Transgenic Nicotiana benthamiana and Arabidopsis thaliana plants that overexpressed VdAK dsRNAs had improved Vd resistance than the wild-type. RT-qPCR results showed that VdAK was also crucial for energy metabolism. Importantly, in an analysis of total small RNAs from Vd strains isolated from the transgenic plants, a small interfering RNA (siRNA) targeting VdAK was identified in transgenic N. benthamiana. Our results demonstrate that HIGS is a promising strategy for efficiently screening pathogenicity/virulence-related genes of Vd and that VdAK is a potential target to control this fungus.

Worldwide, Verticillium wilt is among the major harmful diseases in cotton production, causing substantial reduction in yields. While this disease has been extensively researched at the molecular level of the pathogen, the molecular basis ofV. dahliaehost response association is yet to be thoroughly investigated. In this study, RNA-seq analysis was carried out onV. dahliaeinfected twoGossypium hirsutumL. cultivars, Xinluzao-36 (susceptible) and Zhongzhimian-2 (disease resistant) for 0 h, 24 h, 72 h and 120 h time intervals. Statistical analysis revealed thatV. dahliaeinfection elicited differentially expressed gene responses in the two cotton varieties, but more intensely in the susceptible cultivar than in the resistant cultivars. Data analysis revealed 4241 differentially expressed genes (DEGs) in the LT variety across the three treatment timepoints whereas 7657 in differentially expressed genes (DEGs) in the Vd592 variety across the three treatment timepoints. Six genes were randomly selected for qPCR validation of the RNA-Seq data. Numerous genes encompassed in disease resistance and defense mechanisms were identified. Further, RNA-Seq dataset was utilized in construction of the weighted gene co-expression network and 11 hub genes were identified, that encode for different proteins associated with lignin and immune response, Auxin response factor, cell wall and vascular development, microtubule, Ascorbate transporter, Serine/threonine kinase and Immunity and drought were identified. This significant research will aid in advancing crucial knowledge on virus-host interactions and identify key genes intricate inG. hirsutumL. resistance toV. dahliaeinfection.