BACKGROUND: Cotton is an important economic crop and a host of Liriomyza sativae. Pectin methylesterase (PME)-mediated pectin metabolism plays an indispensable role in multiple biological processes in planta. However, the pleiotropic functions of PME often lead to unpredictable effects on crop resistance to pests. Additionally, whether and how PME affects susceptibility to Liriomyza sativae remain unclear.
RESULTS: Here, we isolated GhPME36, which is located in the cell wall, from upland cotton (Gossypium hirsutum L.). Interestingly, the overexpression of GhPME36 in cotton caused severe susceptibility to Liriomyza sativae but increased leaf biomass in Arabidopsis. Cytological observations revealed that the cell wall was thinner with more demethylesterified pectins in GhPME36-OE cotton leaves than in WT leaves, whereas the soluble sugar content of GhPME36-OE cotton leaf cell walls was accordingly higher; both factors attracted Liriomyza sativae to feed on GhPME36-OE cotton leaves. Metabolomic analysis demonstrated that glucose was significantly differentially accumulated. Transcriptomic analysis further revealed DEGs enriched in glucose metabolic pathways when GhPME36 was overexpressed, suggesting that GhPME36 aggravates susceptibility to Liriomyza sativae by affecting both the structure and components of cell wall biosynthesis. Moreover, GhPME36 interacts with another pectin-modifying enzyme, GhC/VIF1, to maintain the dynamic stability of pectin methyl esterification.
CONCLUSIONS: Taken together, our results reveal the cytological and molecular mechanisms by which GhPME36 aggravates susceptibility to Liriomyza sativae. This study broadens the knowledge of PME function and provides new insights into plant resistance to pests and the safety of genetically modified plants.

Cotton (Gossypium hirsutum L.) is of great economic importance as a cultivated crop in many parts of the world. In addition to being a pillar of the textile industry, cotton and its byproducts are used for livestock feed, seed oil, and other products. Bacillus thuringiensis crystal toxin (Bt) expression in cotton provides effective protection against chewing insects but does not defend plants from piercing/sucking insect pests. With the aim to create transgenic plants with resistance against piercing/sucking pests, we used Agrobacterium-mediated genetic transformation of cotton cultivar Coker 312 to express the Allium sativum leaf agglutinin (ASLA) gene from the phloem-specific rolC promoter. The ASLA transgene was stably inherited and showed Mendelian segregation in the T1 generation. Transgenic lines, expressing the ASLA gene, showed explicit resistance against major sap-sucking pests. Green peach aphid (Myzus persicae Sulzer) choice assays showed that 75% of aphids preferred untransformed cotton plants relative to those expressing the ASLA gene. In detached leaf bioassays, plants expressing ASLA caused 82% aphid mortality and 44-53% reduction in fecundity. Clip cage bioassays with whiteflies (Bemisia tabaci Gennadius) showed 74-82% mortality and 44-60% decrease in fecundity due to ASLA gene expression. In whole plant bioassays, whiteflies showed 77% mortality and a 54% decrease in fecundity on ASLA transgenics. Importantly, we did not observe a negative effect of the ASLA gene on ladybugs (Coccinella septempunctata) that consumed these whiteflies. Together, our findings demonstrate the potential of ASLA-transgenic cotton for providing protection against two devastating insect pests, whiteflies and aphids. The ASLA-transgenic cotton appears promising for direct commercial cultivation besides serving as a potential genetic resource in recombination breeding.

BACKGROUND: Calcium-dependent protein kinase (CDPK) plays a key role in cotton tolerance to abiotic stress. However, its role in cotton heat stress tolerance is not well understood. Here, we characterize the GhCDPK gene family and their expression profiles with the aim of identifying CDPK genes associated with heat stress tolerance.
RESULTS: This study revealed 48 GhCDPK members in the cotton genome, distributed on 18 chromosomes. Tree phylogenetic analysis showed three main clustering groups of the GhCDPKs. Cis-elements revealed many abiotic stress and phytohormone pathways conserved promoter regions. Similarly, analysis of the transcription factor binding sites (TFBDS) in the GhCDPK genes showed many stress and hormone related sites. The expression analysis based on qRT-PCR showed that GhCDPK16 was highly responsive to high-temperature stress. Subsequent protein-protein interactions of GhCDPK16 revealed predictable interaction with ROS generating, calcium binding, and ABA signaling proteins. Overexpression of GhCDPK16 in cotton and Arabidopsis improved thermotolerance by lowering ROS compound buildup. Under heat stress, GhCDPK16 transgenic lines upregulated heat-inducible genes GhHSP70, GHSP17.3, and GhGR1, as demonstrated by qRT-PCR analysis. Contrarily, GhCDPK16 knockout lines in cotton exhibited an increase in ROS accumulation. Furthermore, antioxidant enzyme activity was dramatically boosted in the GhCDPK16-ox transgenic lines.
CONCLUSIONS: The collective findings demonstrated that GhCDPK16 could be a viable gene to enhance thermotolerance in cotton and, therefore, a potential candidate gene for improving heat tolerance in cotton.

Verticillium wilt (VW), caused by the soil-borne plant pathogenic fungus Verticillium dahliae, is a major disease impacting olive crops globally. In view of the lack of effective post-infection treatments, exclusion and avoidance strategies are essential in disease management. Assessing the risks posed by this pathogen is essential to prevent the spread and to ensure selection of suitable sites for new plantations. This study aimed to elucidate the environmental factors driving V. dahliae establishment in the Andalusia region, in southern Spain, an emblematic Mediterranean landscape for olive cultivation. To this end, we explored ecological niche signals for this fungal pathogen by analyzing 62 environmental variables across 1.6 million hectares dedicated to olive and cotton cultivation, using a 15-yr survey data on VW incidence on presence-absence from both olive and cotton fields. To ensure robust identification of ecological niche signals, we employed randomization-based, non-parametric univariate tests to compare presence records with the broader sampling universe (including absence records). Our findings identified key environmental variables that are associated significantly with V. dahliae presence, including temperature range seasonality (including mean diurnal and annual ranges), summer temperature (maximum of the warmest month, mean of the warmest quarter), and moisture and water availability (near-surface humidity, potential evapotranspiration, vapor pressure) as core niche variables for V. dahliae. Our results replicated the pathogen\'s known distribution, identifying the Guadalquivir Valley as a particularly high-risk area in view of its mild winters and distinct rainy seasons, providing new insights into the specific environmental conditions that facilitate the pathogen\'s survival and spread. Furthermore, this study introduces a novel approach to niche modeling that prioritizes variables with consistent effects and significant impact on the presence and distribution of V. dahliae and identifies potential data artifacts. This approach enhances our understanding of ecological requirements in V. dahliae and informs targeted management strategies.

Off-target pesticide drift from cropland is a major source of pesticide exposure to pollinating insects inhabiting crop and wildlands in the lower Mississippi Delta (LMD) in the USA. This study is aimed to develop a drift-reducing pesticide adjuvant that is less/nontoxic to honeybees. Ongoing toxicology experiments with two widely-used insecticides and sodium alginate (SA) pointed out reductions in honeybee mortality compared to an industry standard reference polyacrylamide (PAM). When used as an adjuvant to spray the same insecticides described above, SA did not interfere in killing the target pests. Therefore, SA has been tested as a drift-reducing pesticide adjuvant to protect honeybees. Spray experiments in the lab were carried out in four sets: (i) water only, (ii) water and adjuvant, (iii) water and pesticide, and (iv) water, pesticide and adjuvant. Each set contained 18 treatment combinations to cover the ranges in spray pressure (three), adjuvant dose (three), and spray nozzles (two). The droplet spectrum was analyzed using a P15 image analyzer. Diameters of 10 %, 50 % and 90 % volumes (DV10, DV50, and DV90), droplet velocity, standard deviation and relative span were measured. The drift reduction potential (DRP) of SA was analyzed by (i) dose, (ii) spray pressure, and (iii) nozzle type. The DRP of SA is compared to that of PAM. Additionally, three field experiments were carried out to analyze the efficiency of SA in reducing pesticide drift. The results from our experiments collectively indicate that SA has significant potential in mitigating drift as well as minimizing pesticide toxicity to honeybees.

The principal component of cotton fibers is the cellulose biological macromolecule. However, its highly flammable nature has significantly constrained its utilization in fields where flame retardancy is essential. Herein, in this work, a highly effective binary composite flame retardant coating (APP/MEL-SWCNHs) with ammonium polyphosphate and modified single-walled carbon nanohorns (MEL-SWCNHs) was chemically attached to cotton fabric. With the add-on of 11.3 %, the treated cotton fabric (APP/MEL-SWCNHs)4 exhibited remarkable flame-retardant and self-extinguishing properties. Its LOI value increased to 23.7 ± 0.1 %, and the damage length was significantly reduced from 30.0 ± 0.1 % cm to 7.9 ± 0.1 % cm compared to the pristine cotton fabric. Despite partial carbonization, (APP/MEL-SWCNHs)4 preserved its original structure. Importantly, in the cone calorimeter test, both the pHRR and THR of (APP/MEL-SWCNHs)4 were drastically decreased by 71.8 % and 35.8 %, respectively. The APP/MEL-SWCNHs coating functioned as a flame retardant by inhibiting the emission of flammable volatiles, releasing non-flammable gases, and encouraging the formation of char layer during combustion. Significantly, thermal degradation kinetic analysis revealed that the third-order kinetic equation (O3) was found to have the strongest correlation with (APP/MEL-SWCNHs)4 in both air and N2 atmospheres. The higher activation energy (E) for (APP/MEL-SWCNHs)4 confirmed that incorporating MEL-SWCNHs improved the thermal stability of the char layer.

Flowering in plants is pivotal for initiating and advancing reproductive processes, impacting regional adaptation and crop yield. Despite numerous cloned and identified flowering time genes, research in cotton remains sparse. This study identified GhSWEET42 as a key determinant of the flowering time in cotton, demonstrating that its heterologous expression in Arabidopsis accelerated flowering under LD conditions compared to WT. Transgenic plants exhibited upregulated expression of the flowering inducers AtFT, AtSOC1, AtGI, and AtFKF1, alongside downregulated expression of the repressors AtTSF, AtFLC, and AtRGL2, correlating with the earlier flowering phenotype. GhSWEET42 showed a constitutive expression pattern, with elevated levels in the leaves, petals, and flower buds, and was notably higher in early-maturing cotton varieties. Subcellular localization assays confirmed GhSWEET42\'s presence on the cell membrane. Transcriptome analysis between WT and GhSWEET42-overexpressing Arabidopsis plants revealed 2393 differentially expressed genes (DEGs), spanning 221 biological processes, 93 molecular functions, and 37 cellular components according to Gene Ontology (GO) enrichment analysis. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis categorized the DEGs into metabolism and environmental information processing. These findings enhance the understanding of GhSWEET42\'s function and provide a foundation for elucidating the molecular mechanisms governing flowering time regulation in cotton.

Somatic embryogenesis (SE) is a biotechnological tool used to generate new individuals and is the preferred method for rapid plant regeneration. However, the molecular basis underlying somatic cell regeneration through SE is not yet fully understood, particularly regarding interactions between the proteome and post-translational modifications. Here, we performed association analysis of high-throughput proteomics and phosphoproteomics in three representative samples (non-embryogenic calli, NEC; primary embryogenic calli, PEC; globular embryos, GE) during the initiation of plant regeneration in cotton, a pioneer crop for genetic biotechnology applications. Our results showed that protein accumulation is positively regulated by phosphorylation during SE, as revealed by correlation analyses. Of the 1418 proteins that were differentially accumulated in the proteome and the 1106 phosphoproteins that were differentially regulated in the phosphoproteome, 115 proteins with 229 phosphorylation sites overlapped (co-differential). Furthermore, seven dynamic trajectory patterns of differentially accumulated proteins (DAPs) and the correlated differentially regulated phosphoproteins (DRPPs) pairs with enrichment features were observed. During the initiation of plant regeneration, functional enrichment analysis revealed that the overlapping proteins (DAPs-DRPPs) were considerably enriched in cellular nitrogen metabolism, spliceosome formation, and reproductive structure development. Moreover, 198 DRPPs (387 phosphorylation sites) were specifically regulated at the phosphorylation level and showed four patterns of stage-enriched phosphorylation susceptibility. Furthermore, enrichment annotation analysis revealed that these phosphoproteins were significantly enriched in endosomal transport and nucleus organization processes. During embryogenic differentiation, we identified five DAPs-DRPPs with significantly enriched characteristic patterns. These proteins may play essential roles in transcriptional regulation and signaling events that initiate plant regeneration through protein accumulation and/or phosphorylation modification. This study enriched the understanding of key proteins and their correlated phosphorylation patterns during plant regeneration, and also provided a reference for improving plant regeneration efficiency.

This study reports the earliest directly dated occurrence of archaeological wheat and cotton in the humid forests of West Africa. These are the first archaeobotanical results from the medieval urban center of Ile-Ife, southwestern Nigeria, best known for its famous artworks. Both wheat and cotton likely spread through trans-Saharan trade networks that laid the foundation for later European trade systems. Forty-eight (48) grains of free-threshing wheat (Triticum aestivum/durum) represent the largest assemblage of wheat recovered in sub-Saharan West Africa, which is surprising given that wheat cannot be cultivated locally. Larger quantities of cotton (Gossypium sp.) recovered from late 12th- to early 13th-century CE contexts suggest earlier and more widespread use than wheat. Cotton may have been cultivated and manufactured into cloth locally. The quick adoption of these exotic crops illustrates the active negotiation of prestige through culinary and adornment practices, as well as a high degree of agricultural experimentation.

Cotton fiber (Gossypium hirsutum) serves as an ideal model for investigating the molecular mechanisms of plant cell elongation at the single-cell level. Brassinosteroids (BRs) play a crucial role in regulating plant growth and development. However, the mechanism by which BR influences cotton fiber elongation remains incompletely understood. In this study, we identified EXORDIUM-like (GhEXL3) through transcriptome analysis of fibers from BR-deficient cotton mutant pagoda 1 (pag1) and BRI1-EMS-SUPPRESSOR 1 (GhBES1.4, encoding a central transcription factor of BR signaling) overexpression cotton lines. Knockout of GhEXL3 using CRISPR/Cas9 was found to impede cotton fiber elongation, while its overexpression promoted fiber elongation, suggesting a positive regulatory function for GhEXL3 in fiber elongation. Furthermore, in vitro ovule culture experiments revealed that the overexpression of GhEXL3 partially counteracted the inhibitory effects of brassinazole (BRZ) on cotton fiber elongation, providing additional evidence of GhEXL3 involvement in BR signaling pathways. Moreover, our findings demonstrate that GhBES1.4 directly binds to the E-box (CACGTG) motif in the GhEXL3 promoter region and enhances its transcription. RNA-seq analysis revealed that overexpression of GhEXL3 upregulated the expression of EXPs, XTHs, and other genes associated with fiber cell elongation. Overall, our study contributes to understanding the mechanism by which BR regulates the elongation of cotton fibers through the direct modulation of GhEXL3 expression by GhBES1.4.