Cadmium (Cd) uptake by rice plants and its subsequent movement through food chains pose a notable risk to the health of both plants and humans.  Therefore, understanding the fundamental mechanisms underlying the uptake and movement process is essential.  Through transcriptome analysis, we found that numerous abscisic acid (ABA)-related genes responded to Cd stress.  Exogenous application of ABA significantly reduced Cd accumulation in the shoots and roots of rice plants.  The increased ascorbate peroxidase (APX) enzyme activity, decreased H₂O₂ content, and elevated Cd tolerance index collectively suggest that ABA may mitigate the toxicity of Cd in rice plants.  Further study revealed that exogenous ABA reduced Cd accumulation by regulating Cd transport and cell wall sequestration.  Consistently, mutation of the ABA signaling factor OsABI5 resulted in a significant increase in Cd accumulation in shoots.  Moreover, foliar spraying of ABA during the grain-filling stage significantly reduced Cd accumulation in rice grains, which was attributed mainly to decreased Cd uptake and the inhibition of Cd transportation from roots to shoots and from leaves to grains.  These findings elucidate the underlying mechanisms of the ABA-mediated response to Cd stress in rice and provide a practical reference for coping with Cd pollution in farmlands

Salt stress is a global constraint on agricultural production.  Therefore, the development of salt tolerant plants has become a current research hotspot.  While salt tolerance has evolved more frequently in C₄ grass lineages, few studies have explored the molecular bases underlying salt stress tolerance in the C₄ crop foxtail millet.  In this study, we used a multi-pronged approach spanning the omics analyses of transcriptomes and physiological analysis of the C₃ crop rice and the C₄ model crop foxtail millet to investigate their responses to salt stress.  The results revealed that compared to C₃ rice, C₄ foxtail millet has upregulated abscisic acid (ABA) and notably reduced CK biosynthesis and signaling transduction under salt stress.  Salt stress in C₃ rice plants triggered rapid downregulation of photosynthesis related genes, which was coupled with severely reduced net photosynthetic rates.  In the salt-treated C₃ rice and C₄ foxtail millet, some stress responsive transcription factors (TFs), such as AP2/ERF, WRKY and MYB, underwent strong and distinct transcriptional changes.  Based on a weighted gene co-expression network analysis (WGCNA), the AP2/ERF transcription factor Rice Starch Regulator1 SiRSR1 (Seita.3G044600) was identified as a key regulator of the salt stress response.  To confirm its function, we generated OsRSR1-knockout lines using CRISPR/Cas9 genome editing in rice and its upstream repressor SimiR172a-overexpressing (172a-OE) transgenic plants in foxtail millet, which both showed increased salt tolerance.  Overall, this study not only provides new insights into the convergent regulation of the salt stress responses of foxtail millet and rice, but it also sheds light on the divergent signaling networks between them in response to salt stress

 Drought is one of the major environmental constraints that significantly affects seedling emergence, yield, and quality of Tartary buckwheat, thereby hindering the development of its industry.  However, the molecular mechanisms underlying drought tolerance genes in Tartary buckwheat remain largely unexplored.  Alcohol dehydrogenase (ADH), one of the essential plant proteins, plays a crucial role in growth, development, and stress responses, but its specific role in drought resistance is still unclear.  In this study, we identified an ADH gene FtADH1, using a membership function value of drought tolerance (MFVD) combined with a genome-wide association study (GWAS) and transcriptomic profiles that confers drought tolerance in Tartary buckwheat. Our findings demonstrated that the overexpression of FtADH1 in Arabidopsis and Tartary buckwheat hairy roots enhances drought tolerance by promoting root elongation and mitigating elevated levels of reactive oxygen species (ROS).  Our findings demonstrated that FtADH1 can enhanced tolerance to drought stresses in both Tartary buckwheat and Arabidopsis.  This study identifies the FtADH1 as a new player in affecting ROS level and the stress response of Tartary buckwheat by regulating protective enzyme activities at a high level to scavenge ROS and modulating root growth under drought stress.  Further, we identified proteins interacting with FtADH1 through a prokaryotic expression pull-down assay combined with mass spectrometry, revealing that FtADH1 specifically interacts with the S-adenosyl-L-methionine (SAM) synthetase protein, FtSAMS1.  Overexpression of FtSAMS1 was found to enhance ADH enzymatic activity, leading to increased SAM content in overexpressing Tartary buckwheat hairy roots under water-deficit conditions.  Additionally, FtSAMS1 overexpression induced a drought-resistant phenotype in Arabidopsis and Tartary buckwheat hairy roots under drought stress, revealing the biological function of FtADH1. Evolutionary analysis indicates that ADH1 in Fagopyrum species has undergone significant evolutionary events, including duplication and purifying selection, which may contribute to functional diversification and adaptive advantages such as drought resistance in cultivated buckwheat.  In summary, this study proposes that FtADH1 is a key contributor to drought tolerance, and its interaction with FtSAMS1 holds potential for the development of drought-resistant varieties in Tartary buckwheat and its relative species.

Peanut (Arachis hypogaea L.) bacterial wilt (BW) is a devastating soil-borne disease caused by Ralstonia solanacearum (RS) that poses a significant threat to peanut yield and quality.  Nucleotide-binding leucine-rich repeat (NBS-LRR) proteins are a class of plant-specific immune receptors that recognize pathogen-secreted effector molecules and activate immune responses to resist pathogen infections.  However, the precise functions of AhCN genes (where CN is a class of nucleotide-binding site, leucine-rich repeat receptor (NLR) genes that lack LRR structural domains) in peanut plants are not fully understood.  In this study, a total of 150 AhCN genes were identified and classified into nine subfamilies based on a systematic phylogenetic analysis.  The AhCN genes showed highly conserved structural features, and the promoter cis-elements indicated involvement in plant hormone signaling and defense responses.  After inoculation with RS, the highly resistant peanut variety ‘H108’ significantly outperformed the susceptible variety ‘H107’ based on physiological indicators such as plant height, main stem diameter, and fresh weight, likely due to the inhibition of bacterial proliferation and diffusion in the stem vascular bundle.  AhCN34 was found to be significantly upregulated in ‘H108’ compared to ‘H107’ during plant infection and in response to treatments with each of three plant hormones.  Importantly, AhCN34 overexpression in peanut leaves enhanced their resistance to BW.  These findings demonstrate the great potential of AhCN34 for applications in peanut resistance breeding.  Our identification and characterization of the AhCN genes provide insights into the mechanisms underlying BW resistance in peanut and can inform future research into genetic methods of improving BW resistance in peanut.

Late leaf spot disease (LLS) is one of the most important diseases that cause severe yield losses in peanut.  Peanut has various sources of resistance to LLS, so the identification of resistant quantitative trait loci (QTLs) and the development of related molecular markers are of great importance for the breeding of LLS-resistant peanut.  In this study, 173 individual lines of a recombinant inbred line (RIL) population and the 48K SNP array for genotyping were used to construct a high-density genetic map with 1,475 bin markers and 20 linkage groups.  A total of 11 QTLs were obtained through QTL analysis using the constructed genetic map.  Among them, the stable major QTL qLLS.LG02 was identified on linkage group 2 in all six environments, with the phenotypic variation explained (PVE) ranging from 15.57 to 31.09%.  QTL-seq technology was also employed for a QTL analysis of LLS resistance.  As a result, 14 QTL loci related to LLS resistance were identified using the G prime algorithm.  Notably, the physical positions of qLLS02 and qLLS03 coincided with those of qLLS.LG02 and qLLS.LG03, respectively.  Gene annotation analysis within the 14 QTL intervals from QTL-seq revealed a total of 163 nucleotide-binding site–leucine-rich repeat (NBS-LRR) disease resistance genes, accounting for 22.86% of all resistance (R) genes in the peanut genome and showing a 4.26-fold enrichment with a P-value of 5.19e–57.  Within the QTL region qLLS02 of the resistant parent Mi-2, there was a 5 Mb structural variation (SV) interval containing 81 NBS-LRR genes.  A PCR diagnostic marker was developed, and validation data suggested that this SV might lead to gene deletion or replacement with other genes.  This SV has the potential to enhance peanut resistance to LLS.  The results of this study have significant implications for improving peanut breeding for LLS resistance through the development of associated molecular markers.

Efficient nitrogen management is crucial for developing sustainable strategies aimed at enhancing yield while mitigating negative environmental impacts.  However, research focusing on this aspect in the production of fresh maize is limited.  Therefore, this study analyzed the effects of nitrogen application rates on the yields of 40 sweet and 44 waxy maize varieties at five sites in Zhejiang Province, China, from 2015 to 2019.  The nitrogen application rates were categorized as either relatively high (RHN, >300 kg ha^–1 for sweet maize and >320 kg ha^–1 for waxy maize) or relatively low (RLN).  An increase in nitrogen application rates significantly reduced nitrogen fertilizer partial productivity in both sweet and waxy maize (R²=0.616, P<0.01; R²=0.643, P<0.01), indicating that the optimum nitrogen application rates in this study might be the lowest values (160 kg ha^–1 for sweet maize and 180 kg ha^–1 for waxy maize).  The kernel number per ear of sweet maize had a potentially more significant impact on fresh grain yield than the 1,000-fresh kernel weight under both RLN and RHN.  In waxy maize, 1,000-kernel weight contributed more to fresh grain yield under RLN, while kernel number per ear and 1,000-kernel weight cooperatively affected the yield under RHN.  This study found that sweet maize required taller plant and ear heights, along with an optimal ear–plant height ratio, to enhance dry matter accumulation and increase source size, particularly under RLN, and to ultimately achieve a higher fresh grain yield.  In contrast, a lower ear height and ear–plant height ratio in waxy maize probably contributed more to the greater kernel number and weight under RLN, likely due to a lower ear height which can reduce the distance between sink and source, enabling more efficient photoassimilate allocation to the ear

Dehydrin (DHN) enhances plant resistance to environmental stress by regulating the synthesis of osmotic adjustment substances and scavenging reactive oxygen species.  However, the role of PbDHN3 under salt stress remains unclear.  In this study, salt stress induced high expression of PbDHN3, and the overexpression of PbDHN3 (OE-PbDHN3) enhanced plant growth under salt stress compared to wild-type (WT) plants.  OE-PbDHN3 plants exhibited higher chlorophyll content and root growth capacity than WT plants under salt stress.  Transcriptome analysis revealed that PbDHN3 expression is associated with ethylene signaling pathways.  OE-PbDHN3 transgenic plants substantially influenced ethylene content and the expression of related genes.  Following treatment with exogenous ethephon, the transgenic lines notably inhibited the processes of ethylene synthesis and signaling transduction.  OE-PbDHN3 transgenic lines treated with exogenous ethylene and the ethylene inhibitor 1-MCP demonstrated significant inhibition of ethylene synthesis and signaling transduction, while promoting root development and chlorophyll content.  Under salt stress, OE-PbDHN3 downregulated the expression of ethylene biosynthesis genes PbACO1-like and PbACO2, and signal transduction genes PbEIN3-like during the initial stress phase.  This early regulation mitigated the adverse effects of salt stress on the plants.  These findings demonstrate that PbDHN3 ameliorates the ethylene-mediated plant growth phenotype under salt stress through regulation of ethylene synthesis and signal transduction.

Pear anthracnose, caused by Colletotrichum fructicola, is a devastating disease that seriously affects most pear varieties, compromising their yield and quality.  However, effective control of this pathogen is lacking.  Moreover, the critical resistance responses to C. fructicola in pear are unknown.  To investigate these resistance mechanisms of pear against C. fructicola, transcriptomic and metabolomic analyses were performed on the anthracnose-resistant variety ‘Seli’ and susceptible variety ‘Cuiguan’ after C. fructicola infection.  Differentially expressed genes (DEGs) and differentially accumulated metabolites (DAMs) were mainly involved in metabolism and secondary metabolite synthetic pathways, including α-linoleic acid metabolism, phenylalanine biosynthesis metabolism, unsaturated fatty acids biosynthesis, and biosynthesis of amino acids and their derivatives.  In particular, the accumulation of unsaturated fatty acids (UFAs), amino acids, and their derivatives, such as linoleic acid and its derivatives, lauric acid, N-acetyl-L-glutamic acid, and L-proline, was significantly increased in ‘Seli’ after infection, while the amino acids of oxiglutatione and N-acetyl-L-glutamic acid, as well as the proanthocyanidins, were significantly decreased in ‘Cuiguan’.  These findings suggest that these metabolites may contribute to the differential anthracnose resistance between ‘Seli’ and ‘Cuiguan’.  Overall, our results provid new insights into the regulation of pear anthracnose resistance, which may assist in developing new control strategies and breeding anthracnose-resistant varieties.

While bagged flat peaches gain increasing consumer preference, the bagging process inhibits anthocyanin production and fruit coloration in most peach cultivars.  Certain peach cultivars, including the rapidly expanding ‘Zhongyoupan 9’, demonstrate a light-independent anthocyanin biosynthesis pattern and accumulate anthocyanins under artificial darkness.  The underlying molecular mechanisms, however, remain unelucidated.  This study examined two flat peach varieties: ‘Zhongpan 17’, exhibiting light-dependent anthocyanin biosynthesis, and ‘Zhongyoupan 9’, displaying light-independent anthocyanin biosynthesis.  The pericarp of ‘Zhongyoupan 9’ contained substantially higher anthocyanin levels compared to ‘Zhongpan 17’.  Metabolomic analysis identified the marked increase in anthocyanins, particularly cyanidin-3-O-glucoside, as the primary factor in ‘Zhongyoupan 9’ coloration under artificial darkness.  Transcriptomic analyses revealed that PpHY5 (long hypocotyl 5) showed upregulation in ‘Zhongyoupan 9’ but downregulation in ‘Zhongpan 17’, with its expression pattern correlating positively with color changes in both varieties.  The role of PpHY5 in positively regulating anthocyanin biosynthesis was confirmed through reduced anthocyanin levels in peach fruits treated with a PpHY5 virus-induced gene-silencing construct.  These findings suggest that the PpHY5 regulates red color development in ‘Zhongyoupan 9’ under artificial darkness.

The FW2.2-like (FWL) gene family has been extensively studied across multiple species, demonstrating conserved functions among specific members in organ development, particularly in fruit size regulation.  For species with limited research foundations, such as Chinese jujube, analyzing this gene family is an effective approach to identify candidate genes for fruit size.  This study identified twenty ZjFWL genes and comprehensively analyzed their chromosomal distribution, phylogenetic relationships, gene structure, evolutionary dynamics, expression patterns, and cis-acting elements in their promoters.  Natural variation analysis of the ZjFWL10 sequence identified a significant correlation between a seven-base pair deletion in the conserved domain and jujube fruit size.  To validate the functional implications of the seven-base pair deletion genotype, heterologous overexpression experiments were conducted in tomatoes, generating three overexpression lines.  Comparative analysis with the wild-type demonstrated a significant reduction in fruit size and a notable increase in plant height in the overexpressed lines.  This gene may play a critical role in regulating nutrient partitioning in jujube, ultimately affecting fruit size.  These findings advance our understanding of fruit size regulation mechanisms and offer valuable insights for genetic improvement strategies targeting jujube fruit size.

Black spot, a fungal disease caused by Alternaria brassicae infection, inflicts severe damage on Chinese cabbage.  Through comparative transcriptomic analysis, this study investigated the molecular mechanisms underlying Chinese cabbage’s defense responses to A. brassicae infection.  Notably, we found that the expression of BrERF109 was induced by A. brassicae infection.  Silencing of BrERF109 by an optimized virus-induced gene silencing (VIGS) assay in Chinese cabbage diminished disease resistance, while BrERF109-overexpression in Arabidopsis enhanced it.  Additionally, BrERF109 silencing in Chinese cabbage suppressed indolic glucosinolates gene expression, substantially reducing indolic glucosinolates levels, whereas BrERF109-overexpression in Arabidopsis promoted their accumulation.  BrERF109 directly interacts with the BrIGMT4 promoter, thereby facilitating indolic glucosinolates accumulation and enhancing defense against A. brassicae.  This study elucidates the BrERF109-BrIGMT4 regulatory module in Chinese cabbage’s defense against A. brassicae infection, while providing valuable data for further investigation of plant–A. brassicae interactions.

Mitochondria play a crucial role in plant growth, fertility, and adaptation.  Sugarcane (Saccharum hybrids) represents the world’s primary sugar and energy crop, while S. spontaneum and S. arundinaceum serve as valuable parental germplasm.  Despite their importance, limited research exists regarding the mitochondrial genomes of sugarcane and related species.  This study presents the assembly of mitogenomes from one S. arundinaceum, one S. spontaneum, and five sugarcane cultivars.  Analysis revealed that these mitogenomes, encoding 33 protein-coding genes (PCGs), ranged from 445,578 to 533,662 bp, with GC content between 43.43–43.82%.  The primary structures of S. arundinaceum consisted of three small rings, while S. spontaneum exhibited one ring and one linear structure, and sugarcane displayed two rings; multiple potential conformations emerged due to repeat-mediated recombination.  Additionally, this research developed an intron marker SAnad4i3 capable of species differentiation.  The analysis identified between 540 and 581 C to U RNA editing sites in the PCGs, with six RNA editing sites linked to start or stop codon creation in S. arundinaceum, and five sites each in S. spontaneum and sugarcane hybrids.  Significantly, 30–37 fragments homologous to chloroplast DNA were identified, with S. spontaneum containing the highest number.  These mitogenomes appear to have undergone substantial genomic reorganization and gene transfer events throughout evolution, including the loss of eight PCGs.  This comprehensive study illuminates the genetic diversity and complexity of the Saccharum complex, establishing a foundation for future germplasm identification and evolutionary research.

Several Trichoderma species serve as biocontrol agents in agriculture through their phytopathogen growth inhibition capabilities.  However, the antagonistic mechanism of certain strains primarily operates through direct action.  This study aims to explore an effective strain with comprehensive capabilities and elucidate its practical viability and action mechanism.  Trichoderma gamsii strain TC959, exhibiting robust antagonistic and plant growth-promoting properties, was identified.  The strain directly inhibits plant pathogen through the production of secondary metabolites, siderophores, and chitinase/xylanase, while promotes plant growth via indole-3-acetic acid/gibberellin release.  Additionally, the strain activates induced systemic resistance by enhancing the chlorophyll a/b ratio and jasmonic acid content in pepper seedlings through root colonization, leading to elevated defense-related gene expression, antioxidant enzyme activity, and indole-3-acetic acid/gibberellin production.  These mechanisms collectively enhance disease resistance and promote plant growth.  Moreover, TC959 demonstrates superior resistance to oxidation and chemical fungicides, facilitating strain viability maintenance and ensuring healthy pepper seedling development.  The study concludes that strain TC959 exhibits significant biocontrol potential and comprehensive functions against pepper damping-off disease, warranting further practical applications

The velvet protein family serves as a crucial factor in coordinating development and secondary metabolism in numerous pathogenic fungi.  However, no previous research has examined the function of the velvet protein family in Fusarium oxysporum f. sp. niveum (FON), a pathogen causing a highly destructive disease in watermelon.  In this study, ∆fovel1 and ∆folae1 deletion mutants and ∆fovel1-C and ∆folae1-C corresponding complementation mutants of FON were validated.  Additionally, the phenotypic, biochemical, and virulence effects of the deletion mutants were investigated.  Compared to the wild-type strains, the ∆fovel1 and ∆folae1 mutants exhibited altered mycelial phenotype, reduced conidiation, and decreased production of bikaverin and fusaric acid.  Furthermore, their virulence on watermelon plant roots significantly decreased.  All these alterations in mutants were restored in corresponding complementation strains.  Notably, yeast two-hybrid results demonstrated an interaction between FoVel1 and FoLae1.  This study reveals that FoVEL1 and FoLAE1 play essential roles in secondary metabolism, conidiation, and virulence in FON.  These findings enhance our understanding of the genetic and functional roles of VEL1 and LAE1 in pathogenic fungi.

CRISPR/Cas9-based gene editing research has advanced greatly and shows broad potential for practical application in life sciences, but the Cas9 system is often constrained by the requirement of a protospacer adjacent motif (PAM) at the target site.  While xCas9, a variant derived from Streptococcus pyogenes Cas9 (SpCas9), can recognize a broader range of PAMs, its application in non-model insects is lacking.  In this study, we explored xCas9 activity in gene editing by selecting corazonin (Crz) and the target sites with various PAMs in Locusta migratoria, a destructive insect pest worldwide.  We found that xCas9 could cleave the target site with AG PAM while SpCas9 could not, although xCas9 appeared to have lower activity than SpCas9 at the canonical NGG PAMs.  The heritable homozygous Crz^–/– locust strain was generated by the application of xCas9.  The Crz^–/– strain showed an albino body color, with significantly downregulated expression of several body color-related genes including Pale, Vermilion, Cinnabar, White and β-carotene-binding protein.  In addition, Crz^–/– mutants exhibited significantly reduced expression of Chitin synthase 1, along with a markedly lower chitin content as well as compact and rigid cuticles.  Furthermore, Crz^–/– mutants displayed impaired performance under low-temperature stress, including prolonged lifespan, reduced body weight and smaller body size.  Our results suggest that xCas9 is effective for insect genome editing, and Crz plays essential roles in insect body color, cuticle development and adaptation to low-temperature stress.  The findings of this study extend the application of xCas9 in non-model insects and provide new insights into our understanding of the regulation of insect cuticle development and environmental adaptation.

Maize is a cornerstone of global food security, but it faces increasing challenges from corn aphids, particularly with the widespread adoption of genetically modified Bt maize.  This trend suggests a growing need for sustainable pest control strategies. Methyl salicylate has been proposed as a volatile compound with the potential for managing aphids.  In this study, Y-tube olfactometer and Petri dish dispersal assays showed that methyl salicylate can repel wingless and winged aphids at 0.1 to 1,000 ng μL^–1.  Moreover, at concentrations of 100 and 1,000 ng μL^–1, it was found to attract beneficial insects such as adults and larvae of Harmonia axyridis.  Exposing maize plants to methyl salicylate resulted in a prominent reduction in the number of aphids compared to the control.  In addition, clip cage experiment assays showed that the nymphal development duration was increased, while the adult duration and generation time were reduced, and the reproductive duration and total number of aphid offspring in plants treated with methyl salicylate were dramatically lower than in the control.  Over two years of field trials, methyl salicylate-impregnated alginate beads provided significant reductions in the populations of key aphid species, including Rhopalosiphum padi, Rhopalosiphum maidis, and Aphis gossypii.  Concurrently, there were marked increases in the presence of natural predators such as H. axyridis, Propylaea japonica, Syrphus corollae, and Chrysoperla sinica.  These compelling results underscore the potential of methyl salicylate as a key component in integrated pest management strategies for maize, offering a green alternative to traditional chemical control.

Pluripotent stem cells (PSCs) are useful for developmental and translational research because they have the potential to differentiate into all cell types of an adult individual.  Pigs are one of the most important domestic ungulates, commonly used for food and as bioreactors.  Generating stable pluripotent porcine PSC lines remains challenging.  So far, the pluripotency gene network of porcine PSCs is poorly understood.  Here we found that TBX3-derived induced pluripotent stem cells (iPSCs) closely resemble porcine 4-cell embryos with the capacity of totipotent-like stem cells (TLSCs).  Interestingly, our data suggest that TBX3 facilitates the activation of H3K4me3 methyltransferase, specifically MLL1.  Subsequent investigations revealed that the porcine 4-cell specific gene, MCL1, is a key downstream effector of the TBX3-MLL1 axis.  Together, our study of the TBX3 regulatory network is helpful in the understanding of the totipotency characteristics of pigs.

Skin and hair pigmentation in animals involve intricate regulatory processes.  Circular RNA-microRNA (circRNA-miRNA) networks play vital roles in various biological processes, although their involvement in pigmentation has been underexplored.  This study focused on circKIF27 expression, which differs significantly in melanocytes isolated from white and brown Boer coat-colored skin, yet its function remains unclear.  Here, we investigated the roles of circKIF27 in melanocytes.  In situ hybridization assays demonstrated that circKIF27 is expressed in the cytoplasm of melanocytes.  qRT-PCR results revealed differential expression levels of circKIF27 in various tissues of male and female goats.  Functional analysis showed that circKIF27 overexpression in melanocytes significantly reduces melanin production (P<0.01) and inhibits cell proliferation (P<0.0001).  Bioinformatics analysis identified a putative miR-129-5p binding site on circKIF27, and luciferase reporter assays confirmed their interaction.  Overexpression of miR-129-5p in melanocytes enhances melanin production (P<0.01) and promotes cell proliferation (P<0.05).  Further analysis revealed that TGIF2 possesses two potential miR-129-5p binding sites, and miR-129-5p overexpression in melanocytes significantly inhibits TGIF2 expression (P<0.0001), suggesting a targeted regulatory relationship between these two molecules.  Silencing TGIF2 expression via siRNA-TGIF2 transfection leads to increased melanocyte proliferation (P<0.0001) and increased melanin production (P<0.01).  These findings highlight the involvement of the circRNA-miRNA network in pigmentation, offering new insights into the molecular mechanisms underlying pigmentation and guiding animal hair color breeding strategies.

Influenza A viruses (IAVs) possess variable pathogenic potency causing great economic losses in the poultry industry worldwide and threatening public health.  The control of IAV epidemics desperately necessitates an efficient platform for screening antiviral compounds and evaluating vaccine efficacy.  In this study, we utilized the H9N2 subtype IAV as the working model.  An 11-amino-acid HiBiT tag, derived from NanoLuc luciferase, was incorporated into the flexible linker region of the NS1 protein.  Subsequently, the recombinant HiBiT-tagged virus was rescued.  The recombinant virus exhibited high genetic stability and similar virological characteristics to the parental virus, both in vitro and in vivo.  Particularly importantly, the replication profile of the HiBiT-tagged virus can be easily measured using the Nano-Glo assay system, achieving an efficient screening platform.  Based on this platform, we have developed assays with both convenience and efficiency for screening antiviral reagents, evaluating immunization efficacy, and measuring neutralizing antibodies.

The eutrophication of rivers and lakes is becoming increasingly common, primarily because of pollution from agricultural non-point sources.  We investigated the effects of optimized water and fertilizer treatments on agricultural non-point source pollution in the Nansi Lake basin.  The water heat carbon nitrogen simulator model (WHCNS model) was used to analyze water and nitrogen transport in wheat fields in Nansi Lake basin.  Four water and fertilizer treatments were set up: conventional fertilization and irrigation (CK), reduced controlled-release fertilizer and conventional irrigation (F2W1), an equal amount of controlled-release fertilizer and reduced irrigation (F1W2), and reduced controlled-release fertilizer and reduced irrigation (F2W2).  The results indicated that the replacement of conventional fertilizers with controlled-release fertilizers, combined with reduced irrigation, led to reduced nitrogen loss.  Compared with those of the CK, the cumulative nitrogen leaching and ammonia volatilization of F2W1 were reduced by 8.90 and 41.67%, respectively; under F1W2, the same parameters were reduced by 12.50 and 15.99%, respectively.  Compared with the other treatments, F2W2 significantly reduced nitrogen loss while producing a stable yield.  Compared with those of the CK, ammonia volatilization and nitrogen loss due to leaching were reduced by 29.17 and 27.13%, respectively, water and nitrogen use efficiencies increased by 11.38 and 17.80%, respectively.  F2W2 showed the best performance among the treatments, considering water and fertilizer management.  Our findings highlight the effectiveness of optimizing water and fertilizer application in improving the water and nitrogen use efficiency of wheat, which is of great significance for mitigating nitrogen loss from farmland in the Nansi Lake basin.

Maize/soybean intercropping systems are commonly used in developing countries, but few studies have been performed to elucidate the differences in nutrient efficiency and rhizosphere microbiome, especially when maize is intercropped with different soybean varieties.  In this study, field experiments were conducted to compare the growth and yield of two soybean (Glycine max) varieties, BD2 and YC03-3, and one maize (Zea mays) variety, Huazhen, in mono-cropped and intercropped cultures.  The plant biomass and N content of both crops in BD2/maize intercropping were significantly improved compared to their monoculture, but no such effects were observed in the plants of YC03-3/maize intercropping.  The yield of BD2 intercropped with maize exhibited a 37.5% increment above that of BD2 in monoculture.  Moreover, 19.2–29.1% longer root length of maize and 19.0–39.4% larger root volume of BD2 were observed in BD2/maize intercropping than in monoculture, but no growth advantage was observed in YC03-3/maize intercropping.  Maize showed root avoidance when intercropped with BD2, but space competition when intercropped with YC03-3.  16S rRNA amplicon sequencing showed that compared with the monoculture system, rhizobacteria community composition in BD2/maize intercropping changed more significantly than that of the YC03-3/maize intercropping system.  In BD2/maize intercropping, most of the rhizobacteria community biomarker bacteria of BD2 were positively correlated with plant biomass, as well as plant P and N content.  Maize tended to recruit Rhizobiales and Proteobacteria, which showed positive correlation with plant biomass and N content, respectively, as well as soil available N.  In conclusion, soybean varieties determined the advantages of maize/soybean intercropping through root–root interactions and modification of rhizobacteria communities.  Our insight emphasizes a linkage between root traits and the rhizobacteria community, which shows the importance of optimizing intercropping systems by selection of appropriate crop varieties.

Straw return has demonstrated significant potential for enhancing carbon (C) sequestration and nitrogen (N) uptake while concurrently promoting plant productivity.  However, the specific transport and distribution of C produced by photosynthesis and exogenous N within the rice plant–soil system under straw return remains unclear.  A long-term straw return pot trial experiment was conducted in a double cropping rice system, incorporating treatments of inorganic fertilizer application with straw removal (F), straw burning and ash return with reducing inorganic fertilizers (SBR), and straw return with reducing inorganic fertilizers (SR) to investigate C sequestration and exogenous N uptake using ¹³C pulse and ¹⁵N isotope tracer techniques.  The SR treatment had significantly higher soil ¹³C abundance, by 24.4 and 25.4%, respectively, ¹³C concentrations in aboveground plant parts, by 18.4 and 35.8% respectively, and ¹⁵N concentrations in rice panicles, by 12.8 and 34.3% than the SBR and F treatments.  This enhancement contributed to a higher total organic C concentration and increased rice grain yield in the SR treatment.  Furthermore, the SR treatment had significantly higher photosynthetic C, by 9.8%, which was directly transferred to soil C.  The SR treatment had a higher distribution of photosynthetic C in the leaves and stems, but a lower distribution in the panicle compared to the SBR treatment.  This finding is advantageous for sequestering photosynthetic C into the soil through straw return; conversely, opposite trends were observed in ¹⁵N distribution.  In addition, rice plants in the SR treatment had increased N uptake from urea and soil N sources, enhancing N recovery by 9.2 and 12.5%, respectively, and reducing soil N residues.  Correlation analysis showed that the SR treatment increased the concentrations of ¹³C in leaves and roots while decreasing the ¹⁵N abundance in all rice organs, thereby contributing to an increase in rice yield.  The partial least square path model suggested that the increase in rice yield under the SR treatment was primarily linked to ¹³C accumulation within the rice plant–soil system.  The results suggest that straw return increases the sequestration of photosynthetic C and exogenous N in the rice plant–soil system and increases N utilization efficiency, which subsequently improves both rice and soil productivity.

The increasing frequency of compound extreme events under ongoing climate change threatens global food security.  Compared to individual extreme events, the simultaneous occurrence of multiple extreme events can exacerbate crop yield reductions, yet comprehensive assessments of these compound effects remain limited.  To bridge this gap, we applied a linear mixed-effects model to quantify the impacts of individual extreme events (cold days (CD) and killing degree days (KDD)) and triple compound extreme events (heatwave and low precipitation (HWLP) and hot-dry-windy (HDW)) on the global yields of winter wheat, soybeans, and maize from 1982 to 2016.  Our analysis indicated that regions severely impacted by extreme events (exceeding the 95% threshold) experienced total crop yield losses of more than 9.16, 24.89, 26.69, and 7.12% due to CD, KDD, HWLP, and HDW, respectively.  The adverse effects of compound events were particularly pronounced during critical growth stages.  HWLP results in yield losses of 9.4% for winter wheat and 6.8% for maize per 10 hours of exposure during the heading to harvesting stages, while soybean yields declined by 8.8% per 10 hours during the planting to three-true-leaf stage.  Similarly, KDD caused a 7.4% yield reduction in winter wheat per 10°C day during the heading to harvesting stages, a 9.5% reduction in maize per 10°C day during the planting to jointing stages, and a 3.8% reduction in soybean per 10°C day during the planting to three-true-leaf stages.  These findings underscore the substantial contribution of compound extreme events, which are often overlooked in existing risk assessments, in determining the global yields of major staple crops.