Disrupting starch biosynthesis in soybeans redirects seed carbon partitioning toward soluble sugars, offering a genetic strategy to breed sweeter crops.

Zika virus (ZIKV) can persist in mosquito populations through both horizontal and vertical transmission, and successful ovarian infection is a prerequisite for transgenerational spread. However, the mosquito host factors that regulate ovarian permissiveness and vertical transmission remain poorly characterized. Here, we investigated the role of cytochrome P450 4G15 (CYP4G15) in ZIKV infection and vertical transmission in Aedes albopictus and identified an associated downstream pathway involving SORD-like, a predicted sorbitol dehydrogenase-like gene. Expression profiling revealed that CYP4G15 exhibits distinct tissue-, developmental-, blood-feeding-, and infection-responsive patterns, with transient induction in ovaries during the early phase of ZIKV infection. RNA interference-mediated silencing of CYP4G15, followed by transcriptomic analysis and RT-qPCR validation, identified SORD-like (LOC109415972) as a candidate CYP4G15-associated gene. SORD-like was highly expressed in the fat body, strongly induced after blood feeding, and responsive to ZIKV challenge in ovarian tissues. Functional assays showed that knockdown of SORD-like significantly reduced early ovarian infection rate and viral RNA accumulation, while also decreasing conditional progeny egg-pool positivity rate and minimum egg infection rate, indicating reduced ZIKV RNA detection in progeny egg pools. SORD-like silencing did not alter CYP4G15 transcript levels, whereas CYP4G15 overexpression was associated with increased SORD-like transcript abundance and functionally compensated for the ovarian viral RNA phenotype caused by SORD-like knockdown. Together, these findings support a functional association between CYP4G15, SORD-like, and ovarian ZIKV infection, but do not establish direct transcriptional regulation or a strict linear epistatic relationship. Our findings reveal a CYP4G15–SORD-like-associated metabolic-regulatory pathway that links mosquito physiology to arbovirus persistence. Given that CYP4G-related pathways are often associated with environmental adaptation and insecticide resistance-associated physiology, this pathway may represent an important point of intersection between vector fitness and viral transmission. It may therefore serve as a candidate target for reducing egg-associated ZIKV RNA detection and ZIKV maintenance in Aedes albopictus populations.

The Asian tiger mosquito (Aedes albopictus) is a rapidly expanding invasive species and a global vector of arboviruses. Its transmission depends on the female's blood meal-driven reproductive cycle. Although mosquito PIWI-interacting RNAs (piRNAs) research has predominantly focused on antiviral immunity, their roles in reproduction remain poorly understood. Here we profiled sex-biased piRNA expression in Ae. albopictus and identified a female-enriched piRNA, AalpiRNA-18529. This piRNA accumulates in pre-vitellogenic ovaries and is rapidly downregulated after a blood meal. Gain-of-function experiments showed that AalpiRNA-18529 mimics impair follicle development, reduces fecundity and egg hatching, and disrupts nurse-cell apoptosis required for oocyte maturation. Mechanistically, AalpiRNA-18529 represses Gadd45a, a conserved stress-response regulator. Gadd45a knockdown phenocopies AalpiRNA-18529 gain-of-function, leading to reduced phosphorylation of c-Jun N-terminal kinase (JNK) and thus indicating defective activation of the Gadd45a-JNK apoptotic axis. Collectively, our findings reveal a piRNA-mediated pathway that controls ovarian development in Ae. albopictus during vitellogenesis and highlight the AalpiRNA-18529-Gadd45a-JNK cascade as a potential molecular target for genetic strategies to disrupt mosquito reproduction.

Flavonoids are central to abiotic stress responses, yet the specific signaling roles and evolutionary dynamics of flavonoid biosynthetic intermediates in crop drought adaptation remain elusive. Here, we demonstrate that dihydrokaempferol (DHK) and dihydroquercetin (DHQ), specific intermediate products of the soybean flavanone 3-hydroxylases GmF3H1/2, function as potent signaling molecules that mitigate drought stress. Exogenous DHK/DHQ promoted ABA-dependent stomatal closure and enhanced drought tolerance across diverse dicot species including soybean and tobacco, highlighting a broadly conserved stress-mitigating signaling mechanism. CRISPR/Cas9-generated gmf3hs double mutants exhibited severe drought hypersensitivity due to compromised redox homeostasis and defective stomatal regulation, which could be specifically functionally rescued by DHK/DHQ application. Furthermore, the loss of GmF3H triggered a distinct reproductive trade-off under stress, leading to increased pod initiation but severe filling defects. Multi-omics network analysis revealed extensive rewiring of broader stress-responsive pathways and identified upstream transcription factors, among which GmPHL11 directly binds to and activates the GmF3H1 promoter, and overexpression of GmPHL11 promoted DHK accumulation and enhanced drought stress tolerance in soybean hairy roots. Finally, population genomic analyses demonstrated that the GmF3H1H1 haplotype, which confers superior enzymatic activity and robust root growth under drought stress, might underwent positive selection during soybean domestication. Collectively, our findings redefine the role of GmF3H-derived specific intermediates as potent signaling molecules, providing comprehensive mechanistic and evolutionary insights into flavonoid-mediated drought resilience, developmental trade-offs, and molecular breeding in crops.

The hypocotyl length and elongation is an important characteristic that affect the soybean seedling emergence and photosynthesis. However, the basic genetic mechanism of this feature remains incompletely understood. In this study, the hypocotyl length of four-day germinated soybean seedlings was evaluated before and after 24 h cultivation to assess hypocotyl elongation (HE) in 330 soybean accessions. Five quantitative trait loci (QTLs) that significantly associated with HE trait were detected by genome-wide association study (GWAS) in two models, and they are located on chromosome (Chr.) 2, 3, 11, 15, and 17, respectively. A total of 84 gene models have been found in HE QTLs candidate regions, and with a large proportion enriched in the biological processes of photosynthesis and cell differentiation. A CCCH zinc finger protein gene of GmZFP1 (Glyma.15G262900) was identified as the candidate in the major locus qHE_8 through the analysis of linkage disequilibrium (LD) blocks, gene expression patterns, and natural variation. Three SNPs substantially associated with HE in the GmZFP1 area resulted in 12 haplotypes (Hap 1–12) and four haplotype groups (Hap Ⅰ-Ⅳ). Soybean accessions carrying superior Hap Ⅲ showed significantly higher HE than the soybean lines containing Hap I, and the Hap Ⅲ made up 13.4

Light serves as a primary zeitgeber for regulating circadian rhythms in mosquitoes. As important vector insects, the diurnal Aedes albopictus and the nocturnal Culex quinquefasciatus are not only nuisance biters, but also transmit viruses such as dengue, Zika, and West Nile. We conducted a comparative analysis of the oscillating molecular rhythms in these two mosquito species. Light-blocking paint application experiments reveal species-specific photic pathways. CRISPR/Cas9 knockouts of the Aalopsin1, Cxqopsin6, and Cry1 genes revealed that Ae. albopictus relies principally on the compound eye pathway, whereas Cx. quinquefasciatus depends more on the non-visual ocelli-like pathway for circadian rhythm regulation. In addition, different light conditions can affect the lifespan of adult mosquitoes. These findings reveal the light-regulated circadian rhythm in diurnal and nocturnal vector mosquitoes. Preferential differences in the compound eye versus ocelli-like light-sensing pathways underlie the distinct diurnal activity of Aedes albopictus and the nocturnal activity of Culex quinquefasciatus.

Soil salinity severely threatens soybean productivity worldwide. While transcriptional responses to salt stress are well-documented, the role of post-transcriptional regulation, particularly alternative splicing (AS), remains underexplored. This study combines physiological phenotyping, transcriptome-wide analysis, and molecular genetics to uncover the mechanisms behind the differences in salt tolerance between the salt-sensitive variety Huachun 6 (HC6) and the resistant variety Fiskeby III. Under salt stress, Fiskeby III exhibited superior survival rates and maintained ion homeostasis, as evidenced by a lower Na+/K+ ratio, compared with HC6. Transcriptomic and splicing analysis revealed extensive salt-induced alternative splicing reprogramming. Genes undergoing differential AS were enriched in pathways related to stress response, ion transport, and RNA splicing. Based on the overlap with both differentially expressed genes (DEG) and alternative splicing (DAS) genes under salt stress, a key splicing factor, GmSR34b, was identified as a central regulator of AS under salt stress. Under NaCl stress, the expression of GmSR34b in leaves peaked at 1 h and a salt stress-specific splicing variant was rapidly induced. A comparative analysis showed that the Fiskeby III cultivar prioritized maintenance of the full-length transcript during prolonged stress, whereas the HC6 cultivar accumulated higher levels of the splicing variant. This indicates differences in the regulation of alternative splicing between these two cultivars. Functional validation confirmed that overexpression of GmSR34b in soybean hairy roots inhibited salt tolerance. This study provides novel insights into the molecular mechanisms of salt tolerance in soybean, suggesting potential strategies for breeding resilient crops through the manipulation of splicing regulators.

The mirtron pathway represents a distinct category of noncanonical microRNA (miRNA) biogenesis mechanisms. Current studies suggest that the mirtron pathway may be widely prevalent across various taxa, including animals and plants, but investigation of this pathway has focused mainly on mammals, particularly humans, and the biological functions and emerging roles of several mirtrons in human diseases have been elucidated. In the context of insects, mirtrons have only been comprehensively characterized and preliminarily functionally analyzed in Drosophila . The Asian tiger mosquito, Aedes albopictus , is a highly invasive species and an important vector of arbovirus transmission to humans. Although canonical miRNA function has been studied in depth in mosquitoes, the role of mirtrons in this species remains to be revealed. In this study, we identified and validated 2 novel conventional mirtrons in Ae. albopictus that are precursors of miR‐11900 and miR‐11893. Mirtronic miRNA biogenesis depends on the splicing of introns and cleavage by Dicer but does not necessarily correlate with intron location in host genes. The molecular evolution of mirtrons was analyzed using methods based on host genes and their exon‒intron architecture; the results indicate that mirtronic miRNAs are relatively young and that they may have appeared in Culicinae after the Anophelinae and Culicinae diverged. According to small RNA sequencing (RNA‐seq) and RNA‐seq data on post‐mirtronic miRNA overexpression, mosquito mirtronic miRNAs are present in low abundance, and the absence of typical target genes in Ae. albopictus suggests they are not involved in post‐transcriptional gene regulation. Overall, our results indicate that the emergence of 2 mirtrons in Ae. albopictus is likely due to the formation of Dicer‐recognized secondary structures during the evolution of the intron sequence; these structures are similar to byproducts processed by Dicer, and their abundance is controlled by an alternative adventitious mirtron emergence‐dependent mechanism. Our study identifies for the 1st time mirtrons in insect species distinct from Drosophila melanogaster , provides new insights into mirtron evolution, and provides a reference for the functional analysis of mirtrons.

Pre-mRNA, produced by eukaryotic DNA transcription, undergoes splicing by the spliceosome, which removes introns and joins exons to form mRNA. The spliceosome is a large and highly dynamic molecular machine. Its core components include five small nuclear ribonucleoproteins (snRNPs) and the various spliceosome-related proteins. The conserved Smith (Sm) complex and the Sm-like proteins (LSm) serve as primary components of the snRNPs. Sm proteins are involved in processes such as pre-mRNA splicing and mRNA degradation, which can regulate gene expression, thereby influencing plant growth, development, and stress responses. While 25 Sm proteins have been identified in rice, their specific roles in regulating rice growth and development remain unclear. In this study, we employed the CRISPR/Cas9 system to edit 15 OsSm genes, and 13 mutants were obtained, with mutation rates ranging from 20.83 to 83.87%. In comparison to the wild type (WT), the mutants exhibited dwarfism, reduced tiller numbers, lower seed-setting rates or sterility, and increased susceptibility to diseases. One Sm mutant, ossmf-2, exhibited dwarfism, delayed flowering, and small grains. Through transcriptome analysis, three target genes, OsMRG702, OsRGG2, and OsLA1, were identified. Mutations of the OsSmF protein may lead to the abnormal splicing of these genes and finally lead to the inhibition of growth and development. Our study first edited the OsSm genes and generated a mutant library in rice. Most of the mutants exhibited abnormal growth and development, underscoring the essential roles of OsSm proteins in rice physiology. Furthermore, this work addresses a critical gap in the functional characterization of Sm proteins in rice. The resulting mutant collection offers valuable germplasm resources and lays a theoretical foundation for elucidating the molecular regulatory networks involving spliceosomal components and their target genes in the control of crop growth, development, and reproduction.

BACKGROUND:Mosquitoes with aggressive biting behavior are important disease vectors threatening public health. Armigeres subalbatus, as an emerging arbovirus and filarial disease vector, exhibits aggressive host-seeking behavior and unique breeding preference for contaminated water. However, the molecular mechanisms underlying these biological characteristics remain poorly understood. This study aimed to generate a high-quality genome assembly and characterize the genetic basis of vector competence and environmental adaptation in Ar. subalbatus. METHODS:We sequenced and assembled the Ar. subalbatus genome using Oxford Nanopore long-read sequencing, Illumina short-read sequencing, and Hi-C technology. Comparative genomic analysis was performed to identify gene families related to detoxification, diapause, innate immunity, and sex determination. Gene structure analysis focused on the male-determining factor and its evolutionary relationships with other mosquito vectors. RESULTS:The genome assembly consists of three chromosomes, with a total size of 1.33 Gbp and an N50 of 430.15 Mbp (GenBank assembly: GCA_024139115.2), displaying 99.4% Benchmarking Universal Single-Copy Orthologs (BUSCO) completeness. We identified the gene structure of the male-determining factor (AsuMf) and characterized its evolutionary relationship with other mosquito vectors. The analysis revealed expanded detoxification-related gene families including cytochrome P450s, which may facilitate adaptation to contaminated breeding sites. We characterized 566 putative diapause-related genes that could potentially contribute to geographical expansion, 334 innate immune genes, and 1673 endogenous viral elements, indicating complex virus-host interactions throughout evolution. CONCLUSIONS:Our study provides insights into the molecular basis of vector competence and adaptation in Ar. subalbatus. The expanded detoxification gene families may enable the species to survive in polluted environments, while the identified diapause-related genes could explain its geographical expansion capabilities. These findings establish a foundation for developing novel vector control strategies targeting this emerging disease vector.

BackgroundSoil salinization poses a global threat to agriculture, necessitating strategies to enhance plant salt stress tolerance. Understanding the metabolic and transcriptomic responses of tobacco plants to salt stress is crucial for developing such strategies.ResultsThis study identified 238 up-regulated and 122 down-regulated metabolites in tobacco plants under long-term salt stress. Initial stress stages activated galactose and sucrose metabolic pathways. Chlorophyll synthesis was impacted by decreased 5-aminolevulinic acid production, while proline accumulation helped mitigate cell damage. Metabolite-metabolite correlation analysis revealed significant correlations among metabolites, and enrichment analysis highlighted benzamides, amino acids, fatty acids, and monosaccharides. Transcriptome analysis identified 8,386 differentially expressed genes, with enriched pathways in hormone signaling, photosynthesis, and amino acid metabolism. Integrated analysis confirmed the involvement of sucrose pathway in the salt response, validated by qRT-PCR.ConclusionsThis study provides a comprehensive understanding of the regulatory networks in tobacco during salt stress. The findings lay the groundwork for future research on plant stress responses and the development of salt-tolerant tobacco cultivars.

The seed-coat color and seed size have an impact on both the evolutionary fitness and the grain yield of crops. Soybean is a major oil crop, and the seed-coat color and seed size exhibit natural diversity among the different soybean varieties. Here, we found an R2R3-MYB transcription factor of GmMYB62, which shows a significant increase in expression as the seed-coat color changes from yellow to black in different soybean varieties. The GmMYB62 was specifically highly expressed in reproductive organs, especially in floral organs in soybeans. The GmMYB62 encodes a nuclear protein that contains two MYB domains. In the phylogenetic analysis, the GmMYB62 was relatively conserved after the divergence of the monocots and dicots, and it also grouped with transcriptional repressors of MYBs in anthocyanin synthesis. The GmMYB62 was overexpressed in Arabidopsis and the seeds displayed a pale-brown coat in GmMYB62 overexpression lines, in contrast to the dark-brown seed coat observed in wild-type of Col-0. The anthocyanin content in the GmMYB62 overexpression lines was dramatically reduced when compared to Col-0. Additionally, the seeds in overexpression lines showed shorter lengths, larger widths, and lower thousand-seed weights than those in Col-0. Furthermore, the genes related to anthocyanin synthesis and seed size regulation were investigated, and expression of eight genes that involved in anthocyanin synthesis pathway, like chalcone synthase (CHS), chalcone isomerase (CHI), flavanone 3-hydroxylase (F3H), and anthocyanidin synthase (ANS) were severely inhibited in the GmMYB62 overexpression lines when compared to Col-0. In addition, the ARGOS-LIKE (ARL), B-Type Cyclin 1 (CYCB1), and enhancer of DA1-1 (EOD3), which govern cell expansion and proliferation, were highly expressed in GmMYB62 overexpression lines when compared to Col-0. Overall, this study sheds new light on the control of seed-coat color and seed size by GmMYB62 and provides potentially valuable targets for improving crop seed quality.

Although many long noncoding RNAs have been discovered in plants, little is known about their biological function and mode of action. Here we show that the drought-induced long intergenic noncoding RNA DANA1 interacts with the L1p/L10e family member protein DANA1 -INTERACTING PROTEIN 1 (DIP1) in the cell nucleus of Arabidopsis, and both DANA1 and DIP1 promote plant drought resistance. DANA1 and DIP1 increase histone deacetylase HDA9 binding to the CYP707A1 and CYP707A2 loci. DIP1 further interacts with PWWP3, a member of the PEAT complex that associates with HDA9 and has histone deacetylase activity. Mutation of DANA1 enhances CYP707A1 and CYP707A2 acetylation and expression resulting in impaired drought tolerance, in agreement with dip1 and pwwp3 mutant phenotypes. Our results demonstrate that DANA1 is a positive regulator of drought response and that DANA1 works jointly with the novel chromatin-related factor DIP1 on epigenetic reprogramming of the plant transcriptome during the response to drought.

A major quantitative trait locus (QTL) for the hundred-seed weight (HSW) was identified and confirmed in the two distinct soybean populations, and the target gene GmCYP82C4 underlying this locus was identified that significantly associated with soybean seed weight, and it was selected during the soybean domestication and improvement process. Soybean is a major oil crop for human beings and the seed weight is a crucial goal of soybean breeding. However, only a limited number of target genes underlying the quantitative trait loci (QTLs) controlling seed weight in soybean are known so far. In the present study, six loci associated with hundred-seed weight (HSW) were detected in the first population of 573 soybean breeding lines by genome-wide association study (GWAS), and 64 gene models were predicted in these candidate QTL regions. The QTL qHSW_1 exhibits continuous association signals on chromosome four and was also validated by region association study (RAS) in the second soybean population (409 accessions) with wild, landrace, and cultivar soybean accessions. There were seven genes in qHSW_1 candidate region by linkage disequilibrium (LD) block analysis, and only Glyma.04G035500 (GmCYP82C4) showed specifically higher expression in flowers, pods, and seeds, indicating its crucial role in the soybean seed development. Significant differences in HSW trait were detected when the association panels are genotyped by single-nucleotide polymorphisms (SNPs) in putative GmCYP82C4 promoter region. Eight haplotypes were generated by six SNPs in GmCYP82C4 in the second soybean population, and two superior haplotypes (Hap2 and Hap4) of GmCYP82C4 were detected with average HSW of 18.27 g and 18.38 g, respectively. The genetic diversity of GmCYP82C4 was analyzed in the second soybean population, and GmCYP82C4 was most likely selected during the soybean domestication and improvement process, leading to the highest proportion of Hap2 of GmCYP82C4 both in landrace and cultivar subpopulations. The QTLs and GmCYP82C4 identified in this study provide novel genetic resources for soybean seed weight trait, and the GmCYP82C4 could be used for soybean molecular breeding to develop desirable seed weight in the future.

AbstractAedes albopictusshows a rapid global expansion and dramatic vectorial capacity for various arboviruses. Mosquitoes display distinct sexual dimorphisms, only adult females consume blood meals to complete ovarian follicle development. Therefore, cyclic reproduction in female mosquitoes serves as a foundation for the transmission of numerous disease-causing pathogens. Aedes have an expansion of the piRNA biogenesis genes, indicated that piRNA may play multiple functional roles in mosquitoes. Although the antiviral function of piRNA pathway in mosquitoes has been extensively studied, the role of piRNAs in mosquito reproduction remain to be further understood. In the present study, we first profiled the characteristics of sex-biased piRNAs in adultAe.albopictus. Then, we identified a female biased piRNA (Aalpi18529) in adult females, that was highly expressed in ovaries at blood feeding-dependent termination, and depended on PIWI5 and ago3 mediated biogenesis. Aalpi18529 overexpression suppressed ovarian development, and reduced fertility and fecundity in adult females post-bloodmeal. Furthermore, we demonstrated that Aalpi18529 can effectively repress its direct target, growth arrest and DNA-damage-inducible protein 45a (GADD45A), and eventually regulates ovarian development via theGadd45a-mediated JNK-dependent nurse cell apoptosis pathway. Our study is the first to report an endogenous piRNA, which trigger silencing of an important protein-coding gene by posttranscriptional regulation in mosquitoes, expanding our current understanding of the important and multiple roles of piRNAs in biological processes inAe. albopictus.Author SummaryHere, we conducted high throughput piRNA sequencing and comprehensive analysis of piRNA sex-based expression profiles in adult females and males of Aedes albopictus. Based on several established universal tools for research, we demonstrate an ovary-enriched endogenous piRNA, Aalpi-18529, is involved in the regulation of the apoptosis of nurse cells during vitellogenesis via the GADD45A/phosphorylated JNK (pJNK) axis and ultimately affects ovarian development. In general, uncovering the biological functions of sex-biased piRNAs in Aedes albopictus will enhance the understanding of piRNA roles in mosquito Sexual dimorphism (SD) and will provide provide more information about the high reproductive capacity of Aedes albopictus, which is essential to find alternative control strategies.Classification:Research Reports

Soybean is one of the major oil and economic crops for human beings, and the adequate root length ensures proper growth, development, stress tolerance, and yield in soybean. However, only a limited number of quantitative trait loci (QTLs) and genes governing primary root length (PRL) are currently identified in soybean. In this study, extensive variation of the PRL was observed in 324 soybean accessions in the seedling stage, and the soybean lines belonging landrace or cultivar subpopulation showed higher PRL than that belonging wild subpopulation. Eight single nucleotide polymorphisms (SNPs) of three novel QTLs that significantly associated with PRL were detected by genome-wide association study (GWAS), and they are distributed on chromosome (Chr.) 2, 19, and 20, respectively. The candidate genes were searched in three QTLs candidate regions, and all 22 putative candidate genes detected are in QTL qPRL_1. Four tandem duplications of gibberellin 20 oxidase 1-like (GA20ox1) were detected in the linkage disequilibrium (LD) block in the qPRL_1 region, and only GmGA20ox1 (Glyma.02G136000) showed higher expression in root tissue. A notable disparity in the PRL trait was observed when the association panel was genotyped using the SNP of Chr02_14101363 (C/G), and only the landrace and cultivar subpopulation accessions possessed the CC allele. The variant Chr02_14101363 (C/G) was located within the coding sequence region of the GmGA20ox1 gene and resulted in a nonsynonymous mutation. Eleven haplotypes were generated by eight SNPs in GmGA20ox1 and its promoter region, and soybean accessions carrying the superior haplotype (Hap3) of GmGA20ox1 showed highest average PRL trait and higher expression of GmGA20ox1. The genetic diversity analysis of GmGA20ox1 indicated that it was most likely selected during soybean domestication and improvement process, leading to the highest proportion of Hap3 of GmGA20ox1 both in landrace and cultivar subpopulations. The qPRL_1 and GmGA20ox1 identified in this study provide valuable genetic resources for soybean root traits, and GmGA20ox1 could be applied for soybean molecular breeding to develop desirable root system architecture (RSA) in the future.

Background Aedes albopictus is a major arbovirus vector with small stagnant water containers being its oviposition sites. Mosquitoes search for these sites based on their olfactory cues (odor and moisture emanating from the water at the oviposition site), visual cues (size and color of the site), and gustatory cues (ion and nutrient concentration in that water). The gustatory mechanism through which mosquitoes search for oviposition sites remains unknown. Methods To investigate the role of taste receptors in Ae. albopictus oviposition site selection, we developed a laboratory model. This model assessed mosquito behavior in locating and detecting oviposition sites, using a location index to quantify site preference and detection time to measure response to water presence. We compared oviposition site-searching efficiency between mosquitoes with blocked and unblocked appendages, targeting the taste organs. Transcriptome sequencing was conducted to identify differentially expressed genes between water-exposed and unexposed mosquitoes. CRISPR/Cas9 technology was then employed to generate a mutant strain with a targeted gene knockout. Results There was no significant difference between the blocked and unblocked groups in the location index. In contrast, the detection time of the unblocked group differed significantly from all other groups, including those with blocked foreleg tarsus, midleg tarsus, hindleg tarsus, all tibia, and all tarsus. Transcriptome sequencing analyses of water-exposed and unexposed mosquitoes revealed that the taste-related gene gustatory receptor 11(gr11) was differentially expressed. This gene was knocked out with CRISPR/Cas9 technology to generate a pure mutant strain with 2- and 4-bp deletions, which exhibited a significantly longer detection time than the wild-type strain. Conclusions This study reveals the role of Ae. albopictus gr11 in water detection at oviposition sites, thereby providing a theoretical basis and scientific guidelines for managing the breeding sites of these mosquitoes.

The circadian system plays a pivotal role in facilitating the ability of crop plants to respond and adapt to fluctuations in their immediate environment effectively. Despite the increasing comprehension of PSEUDO-RESPONSE REGULATORs and their involvement in the regulation of diverse biological processes, including circadian rhythms, photoperiodic control of flowering, and responses to abiotic stress, the transcriptional networks associated with these factors in soybean (Glycine max (L.) Merr.) remain incompletely characterized. In this study, we provide empirical evidence highlighting the significance of GmPRR3b as a crucial mediator in regulating the circadian clock, drought stress response, and abscisic acid (ABA) signaling pathway in soybeans. A comprehensive analysis of DNA affinity purification sequencing and transcriptome data identified 795 putative target genes directly regulated by GmPRR3b. Among them, a total of 570 exhibited a significant correlation with the response to drought, and eight genes were involved in both the biosynthesis and signaling pathways of ABA. Notably, GmPRR3b played a pivotal role in the negative regulation of the drought response in soybeans by suppressing the expression of abscisic acid-responsive element-binding factor 3 (GmABF3). Additionally, the overexpression of GmABF3 exhibited an increased ability to tolerate drought conditions, and it also restored the hypersensitive phenotype of the GmPRR3b overexpressor. Consistently, studies on the manipulation of GmPRR3b gene expression and genome editing in plants revealed contrasting reactions to drought stress. The findings of our study collectively provide compelling evidence that emphasizes the significant contribution of the GmPRR3b-GmABF3 module in enhancing drought tolerance in soybean plants. Moreover, the transcriptional network of GmPRR3b provides valuable insights into the intricate interactions between this gene and the fundamental biological processes associated with plant adaptation to diverse environmental conditions.

The main vectors of Zika virus (ZIKV) and dengue virus (DENV) are Aedes aegypti and Ae. albopictus, with Ae. aegypti being more competent. However, the underlying mechanisms remain unclear. Here, we find Ae. albopictus shows comparable vector competence to ZIKV/DENV with Ae. aegypti by blood-feeding after antibiotic treatment or intrathoracic injection. This suggests that midgut microbiota can influence vector competence. Enterobacter hormaechei_B17 (Eh_B17) is isolated from field-collected Ae. albopictus and conferred resistance to ZIKV/DENV infection in Ae. aegypti after gut-transplantation. Sphingosine, a metabolite secreted by Eh_B17, effectively suppresses ZIKV infection in both Ae. aegypti and cell cultures by blocking viral entry during the fusion step, with an IC50 of approximately 10 mu M. A field survey reveals that Eh_B17 preferentially colonizes Ae. albopictus compared to Ae. aegypti. And field Ae. albopictus positive for Eh_B17 are more resistant to ZIKV infection. These findings underscore the potential of gut symbiotic bacteria, such as Eh_B17, to modulate the arbovirus vector competence of Aedes mosquitoes. As a natural antiviral agent, Eh_B17 holds promise as a potential candidate for blocking ZIKV/DENV transmission.

Identification and characterization of soybean germplasm and gene(s)/allele(s) for salt tolerance is an effective way to develop improved varieties for saline soils. Previous studies identified GmCHX1 (Glyma03g32900) as a major salt tolerance gene in soybean, and two main functional variations were found in the promoter region (148/150 bp insertion) and the third exon with a retrotransposon insertion (3.78 kb). In the current study, we identified four salt-tolerant soybean lines, including PI 483460B (Glycine soja), carrying the previously identified salt-sensitive variations at GmCHX1, suggesting new gene(s) or new functional allele(s) of GmCHX1 in these soybean lines. Subsequently, we conducted quantitative trait locus (QTL) mapping in a recombinant-inbred line population (Williams 82 (salt-sensitive) × PI 483460B) to identify the new salt tolerance loci/alleles. A new locus, qSalt_Gm18, was mapped on chromosome 18 associated with leaf scorch score. Another major QTL, qSalt_Gm03, was identified to be associated with chlorophyll content ratio and leaf scorch score in the same chromosomal region of GmCHX1 on chromosome 3. Novel variations in a STRE (stress response element) cis-element in the promoter region of GmCHX1 were found to regulate the salt-inducible expression of the gene in these four newly identified salt-tolerant lines including PI 483460B. This new allele of GmCHX1 with salt-inducible expression pattern provides an energy cost efficient (conditional gene expression) strategy to protect soybean yield in saline soils without yield penalty under non-stress conditions. Our results suggest that there might be no other major salt tolerance locus similar to GmCHX1 in soybean germplasm, and further improvement of salt tolerance in soybean may rely on gene-editing techniques instead of looking for natural variations.