As a vital economic crop, grapes suffer substantial postharvest losses due to anthracnose infections. Traditional fungicides, however, pose escalating concerns regarding environmental contamination and the emergence of pathogen resistance, underscoring the urgency for eco-friendly alternatives. This study evaluates the efficacy of quercetin, a plant-derived flavonoid, as a sustainable alternative for enhancing postharvest anthracnose resistance in grape berries. Foliar application of quercetin during the ripening stage significantly reduced the diameter of anthracnose lesions following inoculation. Physiological assays further revealed that quercetin treatment upregulated activities of antioxidant and defense-related enzymes, concomitant with increased accumulation of phenolic compounds and flavonoids. Transcriptomic profiling identified 1733 differentially expressed genes (DEGs), which were predominantly enriched in pathways related to phenylpropanoid and flavonoid biosynthesis, hormone signaling, and glutathione metabolism. Metabolomic profiling revealed 84 differentially abundant metabolites (DAMs), with significant upregulation of flavonoids and phenolic acids. Integrated multi-omics analysis highlighted a dual regulatory role of quercetin: activating phenylpropanoid and flavonoid biosynthetic pathways to enhance accumulation of resistant metabolites, while improving glutathione-mediated detoxification and plant-pathogen interaction signaling cascades. These results elucidate the mechanism by which quercetin mitigates anthracnose via coordinated transcriptional reprogramming and metabolic remodeling, thereby presenting a sustainable, multi-target strategy for postharvest disease management in grapes, reducing reliance on synthetic fungicides while maintaining fruit quality.

Tomato gray mold, caused by Botrytis cinerea, results in significant postharvest losses, necessitating the development of environmentally friendly biocontrol strategies. This study isolated Bacillus amyloliquefaciens strain F028 from tomato rhizosphere and evaluated its biocontrol potential and underlying mechanisms against gray mold. Strain F028 exhibited strong in vitro antagonism against B. cinerea, causing plasma membrane damage. Whole-genome sequencing revealed multiple biosynthetic gene clusters for antimicrobial compounds. In vivo assays on tomato fruit demonstrated that F028 significantly reduced lesion development and pathogen colonization. Molecular analyses indicated that F028 downregulated key fungal virulence genes and activated host defense responses, including upregulation of salicylic acid and jasmonic acid-pathway genes and enhanced antioxidant enzyme activities. Furthermore, volatile organic compounds (VOCs) emitted by F028 strongly inhibited B. cinerea growth both in vitro and on fruit. GC-MS analysis identified 26 VOCs, with 2-undecanol and 2-decanol showing the highest antifungal activity. In conclusion, B. amyloliquefaciens F028 controls gray mold through a multi-layered strategy involving direct antifungal activity, suppression of pathogen virulence, induction of host immunity, and VOC-mediated inhibition, highlighting its promise as a sustainable biocontrol agent.

Wheat (Triticum aestivum L.) is the widest cultivated crop in the world. Abiotic stress, such as drought and high salinity, dramatically impacts the growth and development of wheat and leads to remarkable yield loss. Understanding the underlying mechanisms of abiotic stress tolerance is of great importance to develop high yield varieties with wide adaptability. Ubiquitination is a major type of post-translational modification in eukaryotes. The plant U-Box (PUB) protein is the smallest family in the E3 ligase superfamily, and involved in the responses to various environmental stimuli. Currently, TaPUB57 has been cloned from wheat. It was induced by multiple abiotic stresses and phytohormone. Its ectopic expression increased grain size and drought tolerance, but caused hypersensitive to salt stress in rice. TaPUB57 interacted with and ubiquitinated TaEXPB3. Constitutive expression of TaEXPB3 resulted in small grain size and remarkably enhanced salt tolerance. Moreover, TaPUB57/TaEXPB3 co-expressing rice plants exhibited phenotypes of salt sensitivity and larger grain size relative to TaEXPB3 transgenic lines. Therefore, it is speculated that TaPUB57 acts on grain size and the salt tolerance by ubiquitinating TaEXPB3.

Galactinol synthase (GolS), a crucial enzyme in the synthesis pathway of raffinose family oligosaccharides (RFOs), plays a vital role in plants against abiotic stress. In previous experiments, VvGolS3 cloned from the grape variety 'Zuoyouhong' exhibited cold-inducible expression. However, its specific function and regulatory mechanism remained largely unknown. Additionally, our research has shown that the transcription factor VvDREB2A was involved in grape response to cold stress, yet its molecular interaction with VvGolS3 required further investigation. In this study, it was observed that the expression level of VvGolS3 was significantly higher in cold-resistant grape varieties compared to sensitive ones. Overexpression of VvGolS3 in transgenic Arabidopsis resulted in enhanced cold tolerance, as evidenced by reduced levels of reactive oxygen species (ROS) and increased RFOs content under cold stress conditions. Transient overexpression of VvGolS3 in grapevine leaves also conferred enhanced cold tolerance. Furthermore, overexpressing-VvDREB2A grape calli showed increased cold tolerance and higher levels of RFOs. Similarly, transient overexpression of VvDREB2A in grapevine leaves enhanced cold tolerance. Through yeast one-hybrid (Y1H), electrophoretic mobility shift assay (EMSA), and dual-luciferase reporter system, it was confirmed that VvDREB2A directly binds to the VvGolS3 promoter and promotes its expression. Collectively, these findings indicate that VvDREB2A directly targetes and regulates the expression of VvGolS3, thereby promoting the accumulation of RFOs and enhancing antioxidant enzymes activity, and ultimately improving cold stress tolerance. This study provides the first mechanistic insight into VvGolS3-mediated cold resistance and identifies a novel genetic module (VvDREB2A-VvGolS3) for molecular breeding of cold-tolerant grape cultivars.

Basic leucine zipper (bZIP) transcription factors serve as crucial regulators in plants' response to abiotic stress; however, its function in grapevine heat tolerance is still largely unknown. Here, we undertook a comprehensive investigation of grape genome, leading to the identification of 65 VvbZIP genes, among which 16 VvbZIPs were significantly induced under heat stress. Overexpression of VvbZIP36 enhanced heat tolerance in grape calli, while virus-induced gene silencing (VIGS) of VvbZIP36 reflected thermal sensitivity. Additionally, we examined the metabolomic and transcriptomic profiles of grape seedlings, which showed that grapes exhibited increased accumulation of flavonoids, coinciding with the significantly induced expression of the VvFLS (Flavanol synthase) gene under heat stress. Overexpression of VvFLS also improved the heat tolerance in grape calli by scavenging reactive oxygen species (ROS). A yeast one-hybrid assay (Y1H) demonstrated that VvbZIP36 was capable of specifically activating the promoter of VvFLS, which was subsequently confirmed through a luciferase reporter assay. Furthermore, the overexpression of VvbZIP36 resulted in enhanced quercetin content, while the exogenous application of quercetin improved heat tolerance of grape. Collectively, our findings suggest that VvbZIP36 binds to the VvFLS promoter, thereby enhancing heat stress tolerance by increasing quercetin production and mitigating oxidative damage in grapes.

High temperature reduces anthocyanin accumulation in various horticultural plants. However, the molecular mechanisms underlying the high-temperature-induced reduction of anthocyanin in grape (Vitis vinifera) remain poorly understood. In this study, VvMYB44-1 was identified as a transcriptional repressor of anthocyanin biosynthesis in grape berries, and its gene expression was strongly induced by high-temperature treatment. Overexpression of VvMYB44-1 inhibited anthocyanin accumulation in both grape berries and tobacco (Nicotiana tabacum) by repressing the transcription of the anthocyanin biosynthesis genes dihydroflavonol-4-reductase (VvDFR) and UDP-glucose flavonoid-3-O-glucosyltransferase (VvUFGT). Furthermore, the interaction between VvMYB44-1 and VvWDR2 competitively inhibited the formation of the MYB-bHLH-WD40 (MBW) activation complex and weakened the transcriptional activity of the complex, thereby decreasing anthocyanin accumulation. Additionally, VvMYB44-1 facilitated cytokinin (CK) accumulation by upregulating the expression of the CK synthesis gene lonely guy 8 (VvLOG8) and inhibiting the CK degradation gene CK oxidase 4(VvCKX4), thus contributing to CK-mediated anthocyanin inhibition in grape berries. Moreover, the inhibitory effect of VvMYB44-1 on anthocyanin biosynthesis and its downstream target genes was weakened with the deletion of the ethylene-responsive element binding factor-associated amphiphilic repression (EAR) motif, indicating that the EAR motif is indispensable for the inhibitory effect of VvMYB44-1 on anthocyanin biosynthesis in grapes. These results provide insights into the regulatory network of VvMYB44-1 in high-temperature-mediated anthocyanin biosynthesis in grapes. Transcription factor VvMYB44-1 negatively regulates high-temperature-mediated anthocyanin biosynthesis in grapevine.

This study investigated the physicochemical characteristics and bacterial spoilage of refrigerated oysters from five Chinese provinces, alongside evaluating perilla leaf extract (PLE) as a natural preservative. A total of 62 strains of protease-producing bacteria, predominantly Pseudoalteromonas, Shewanella, and regionally Vibrio, were identified across samples, with serine proteases and metalloproteases as the dominant secreted enzymes, accelerating oyster spoilage. Metagenomic analysis revealed that PLE treatment reduced bacterial diversity, shifting dominant genera from spoilage-associated Pseudoalteromonas and Shewanella to Pseudomonas and Marinomonas. PLE effectively inhibited protein degradation, maintaining higher protein content compared to the control, while lower amino acid nitrogen levels on the seventh day of refrigeration. Furthermore, PLE suppressed the growth of 53 isolated protease-producing strains at 24 h and inhibited their extracellular protease activity, particularly metalloproteases and serine proteases. These findings demonstrate that protease-producing bacteria and their enzymes critically contribute to oyster spoilage, while PLE serves as a sustainable preservative by mitigating microbial deterioration and protease-driven quality loss. The results highlight PLE's dual antimicrobial and protease-inhibiting properties, offering an eco-friendly strategy to enhance seafood shelf life and meet consumer demand for natural preservation solutions.

Grapes are one of the important fruit crops widely cultivated in the world, with high nutritional and economic value. However, with the intensification of global warming, extreme low temperature has seriously affected the development of the grape industry. Quercetin is a highly antioxidant active substance that can enhance the tolerance of plants to external environmental stress, but its function and mechanism in response to low-temperature stress in grapes are still unclear. Here, we found that grapes accumulate more quercetin under low-temperature stress, and exogenous quercetin can significantly improve the cold resistance of grapes. The key quercetin synthesis gene VvFLS1 (flavanol synthase 1) is up-regulated after low-temperature treatment, and overexpression of VvFLS1 increases quercetin content and enhances the cold resistance of grape. Yeast one-hybrid and dual luciferase reporter systems demonstrate that VvbZIP22 (basic-leucine zipper 22) directly binds to the VvFLS1 promoter, and VvbZIP22 has cold-induced expression characteristics. Overexpression of VvbZIP22 significantly improves the cold resistance of grape. The above results indicate that quercetin plays an important role in the response of grapes to low-temperature stress. Under low temperature, VvbZIP22 can mediate quercetin synthesis through regulating VvFLS1, alleviate oxidative damage, and improve the cold resistance of grapes.

BACKGROUND:Bacillus inaquosorum strains is widely recognized for their plant-growth-promoting and biocontrol capabilities, yet their roles in protease production remain unclear. The present study aimed to comprehensively assess the protease-producing performance of B. inaquosorum strain E1-8, at the same time as exploring the novel application of agricultural Bacillus proteases in the preparation of protein hydrolysates for fresh-cut fruits preservation. RESULTS:First, genomic sequencing revealed the diversity of E1-8 proteases, indicating 15 putative extracellular proteases. Subsequently, the fermentation conditions for E1-8 protease production were optimized, with sweet potato powder and soybean meal identified as the most suitable carbon and nitrogen sources, respectively, resulting in a maximum protease activity of 321.48 U mL-1. Upon culturing the strain under these optimized conditions, only an S8 family serine protease and an M48 family metalloprotease were revealed by secretomic analysis and protease inhibitor assays. Additionally, the optimal protease conditions for generating protein hydrolysates from soy, pea, fish and porcine proteins were determined. The molecular weight of the hydrolysates primarily ranged from 2000 to 180 Da, with a total of 17 amino acids identified. The application of these hydrolysates demonstrated a 2,2-diphenyl-1-picrylhydrazyl (i.e. DPPH) scavenging activity ranging from 58.64% to 84.12%, significantly reducing of the melting peaks and the freezing points. Furthermore, the browning index of apple slices stored at 4 °C decreased by 14.81% to 22.15% on the second day, and similar effects were observed in fresh-cut banana stored at 4 °C for 7 days. CONCLUSION:The protein hydrolysates obtained exhibit remarkable antioxidant, antifreeze and anti-browning properties for fresh-cut fruits. © 2024 Society of Chemical Industry.

The booming mudflat aquaculture poses an accumulation of organic matter and a certain environmental threat. Protease-producing bacteria are key players in regulating the nitrogen content in ecosystems. However, knowledge of the diversity of protease-producing bacteria in coastal mudflats is limited. This study investigated the bacterial diversity in the coastal mudflat, especially protease-producing bacteria and their extracellular proteases, by using culture-independent methods and culture-dependent methods. The clam aquaculture area exhibited a higher concentration of carbon, nitrogen, and phosphorus when compared with the non-clam area, and a lower richness and diversity of bacterial community when compared with the clam naturally growing area. The major classes in the coastal mud samples were Bacteroidia, Gammaproteobacteria, and Alphaproteobacteria. The Bacillus-like bacterial community was the dominant cultivated protease-producing group, accounting for 52.94% in the non-clam area, 30.77% in the clam naturally growing area, and 50% in the clam aquaculture area, respectively. Additionally, serine protease and metalloprotease were the principal extracellular protease of the isolated coastal bacteria. These findings shed light on the understanding of the microbes involved in organic nitrogen degradation in coastal mudflats and lays a foundation for the development of novel protease-producing bacterial agents for coastal mudflat purification.

Low temperatures restrict the growth of the grapevine industry. The DREB transcription factors are involved in the abiotic stress response. Here, we isolated the VvDREB2A gene from Vitis vinifera cultivar ‘Zuoyouhong’ tissue culture seedlings. The full-length VvDREB2A cDNA was 1068 bp, encoding 355 amino acids, which contained an AP2 conserved domain belonging to the AP2 family. Using transient expression in leaves of tobacco, VvDREB2A was localized to the nucleus, and it potentiated transcriptional activity in yeasts. Expression analysis revealed that VvDREB2A was expressed in various grapevine tissues, with the highest expression in leaves. VvDREB2A was induced by cold and the stress-signaling molecules H2S, nitric oxide, and abscisic acid. Furthermore, VvDREB2A-overexpressing Arabidopsis was generated to analyze its function. Under cold stress, the Arabidopsis overexpressing lines exhibited better growth and higher survival rates than the wild type. The content of oxygen free radicals, hydrogen peroxide, and malondialdehyde decreased, and antioxidant enzyme activities were enhanced. The content of raffinose family oligosaccharides (RFO) also increased in the VvDREB2A-overexpressing lines. Moreover, the expression of cold stress-related genes (COR15A, COR27, COR6.6, and RD29A) was also enhanced. Taken together, as a transcription factor, VvDREB2A improves plants resistance to cold stress by scavenging reactive oxygen species, increasing the RFO amount, and inducing cold stress-related gene expression levels.

Soil salinization is one of the critical adverse environmental factors affecting agricultural crop growth and yield, and rational use of growth-promoting rhizotrophic bacteria is an effective way to improve and amend salinized soil. In the present study, two halotolerant bacterial strains C8 and B4 were isolated from saline soil in Dongying city, Shandong province. Through morphological observation, physiological and biochemical tests combined with 16S rDNA and gyrB gene sequence analysis, C8 and B4 were identified as Microbacterium oxydans and Stenotrophomonas maltophilia respectively. The results on the LB medium with NaCl showed that the C8 tolerated 6% NaCl, and had the abilities of dissolving potassium, organic and inorganic phosphorus as well as producing auxin. Strain B4 tolerated 8% NaCl, was capable of solubilizing organic phosphorus and producing auxin. Using tomato as material, the effects and mechanism of sole inoculation with C8 or B4 and simultaneous inoculation of C8 and B4 on the seed germination and plant growth were investigated under salt stress. The results demonstrated that sole C8 or B4 inoculation and the simultaneous inoculation of C8 and B4 significantly promoted tomato seed germination and seedling growth, increased catalase（CAT）and peroxidase（POD）activities, and up-regulated the expressions of catalase gene CAT1 and CAT2. C8and B4 application enhanced plant K + content, but reduced Na + content and Na + /K + , and up-regulated the expression of vacuolar membrane Na + /H + antiporter gene NHX1 and NHX3. Simultaneous application of C8 and B4 showed synergistic effect. The above results indicate that C8 and B4enhance antioxidant capacity, maintain ion homeostasis in plants by regulating the expression of antioxidant enzyme and Na + transporter genes,thus improve plant salt tolerance.

Soil salinization is a major factor seriously limiting crop growth and yield.Using soil beneficial microorganisms for saline field amendment is the reliable and sustainable approach.Two halotolerant bacteria strains,Microbacterium oxydans C8 and Stenotrophomonas maltophilia B4,were isolated from saline field in our previous study.Strain C8 has the ability to dissolve potassium,organic and inorganic phosphorus as well as produce auxin.Strain B4 is capable of solubilizing organic phosphorus and producing auxin.In this paper,the effect of mixed culture of C8 and B4 on their functions was studied.The results showed that the mixed culture of C8 and B4 demonstrated significantly stronger ability of dissolving potassium,phosphorus and produce auxin than a single strain.Orthogonal experiments and response surface test were used to optimize the fermentation medium and condition of C8 and B4 mixture,the results showed that the optimized fermentation medium was as follows:glucose 10 g/L,yeast extract 10 g/L,sodium chloride 4.5 g/L.The optimized fermentation conditions were:pH 7.4,temperature 28.8℃,rotating speed 129 r/min,inoculation amount 2%,the liquid content 20%and culture time 23 h.Using tobacco as material,the growth-promoting effects of compound bioinoculant of C8 and B4 were studied.The results showed that,the compound bioinoculant significantly prompted plant growth under salt stress.

The widespread and indiscriminate disposal of plastics inevitably leads to environmental pollution. The secondary pollution by current landfill and incineration methods causes serious damage to the atmosphere, land, and rivers.

Soil proteinase and proteinase-producing microbial community are closely associated with soil fertility and soil health. Sea rice has been planted in the coastal beach of Jiaozhou Bay, China, in an effort to transform saline-alkali soil into arable land. However, the knowledge regarding the bacterial degradation of organic nitrogen in sea rice soils is limited. This study aims to investigate the physicochemical characteristics and enzymatic activities of the sea rice soils, as well as the microbial communities by both the Illumina sequencing-based culture-independent technology and culture-dependent methods. Sea rice soils exhibited a lower salinity and higher organic matter content and proteinase activity, as well as an increase in both the richness and diversity of the proteinase-producing bacterial community, compared to the adjacent non-rice soils. The Proteobacteria phylum and the Gammaproteobacteria class were dominant in sea rice soils, showing higher abundance than in the reference soils. The Planococcus genus and Bacillus-like bacterial communities were abundant in the cultivable proteinase-producing bacteria isolated from sea rice soils. Furthermore, a significant proportion of the extracellular proteinase produced by the isolated soil bacteria consisted of serine proteinases and metalloproteinases. These findings provided new insights into the degradation of soil organic nitrogen in coastal agricultural regions.

With the intensification of global warming, extreme weather events have occurred more frequently, among which cold stress has become one of the major environmental factors that restrict global crop yield and production. Multiple long noncoding RNAs (lncRNAs) have been predicted or recognized in the plant response to cold stress, however, the molecular biological functions of most of these RNAs are still poorly understood. Here, we identified a novel lncRNA, COLD INDUCED lncRNA 1 (CIL1), as a positive regulator of the plant response to cold stress in Arabidopsis. CIL1 was significantly induced when the plant was exposed to cold stress. Moreover, knockdown mutants showed more sensitivity to cold stress than the wild type did, accompanied by an increased content of endogenous ROS (reactive oxygen species) and reduced osmoregulatory substances. Genome-wide transcriptome analysis indicated that 256 genes were downregulated and 34 genes were upregulated in cil1 mutants under cold stress, which were mainly involved in hormone signal transduction, ROS homeostasis and glucose metabolism. Our study implies that CIL1 has a positive effect on the plant response to cold stress by regulating the expression of multiple stress-related genes during the seedling stage.

传承发展提升农耕文明和推动现代农业绿色发展,是高等农业院校弘扬社会主义核心价值观,落实立德树人根本任务,培养创新型人才的重要任务之一.植物生理学的课程内涵与农耕文明和农业绿色发展紧密相关.为此,构建了"以助力农业绿色发展为引领,以传承发展提升农耕文明为依托,以弘扬科学家精神为载体,以培养绿色创新能力为目标"的课程思政育人体系.围绕课程知识点深度挖掘课程思政元素,结合现代信息技术,创建"教师发展平台、学生成长平台、信息拓展平台和实践创新平台"相结合的多元化教学模式,通过课程思政的教学设计和实施,为培养有"学农、爱农、兴农"情怀的创新型现代农业科技人才提供有力支撑.

为获得优质的解磷促生菌种资源,助力农业生产减肥增效,对山东省高密市方市乡烟草根际土壤解磷菌进行筛选,将筛选得到的高效解磷菌3P29进行分子生物学鉴定,并研究其解磷性能及促生能力.菌株3P29鉴定为皮特不动杆菌,其对卵磷脂的转化量为13.38 mg/L,磷酸钙的转化量为19.83 mg/L,具有高效分解有机磷及无机磷的功能.烟草盆栽试验表明,烟草主根变长且有更多侧根产生,从而增强根对营养元素的吸收,烟草叶片全氮磷钾含量在无磷营养液条件下提高了71％、49％和134％,可为解磷菌肥的研发与利用提供优良菌株资源.

Ubiquitination-mediated protein degradation in both the 26S proteasome and vacuole is an important process in abscisic acid (ABA) signaling. However, the role of deubiquitination in this process remains elusive. Here, we demonstrate that two deubiquitinating enzymes (DUBs), ubiquitin-specific protease 12 (UBP12) and UBP13, modulate ABA signaling and drought tolerance by deubiquitinating and stabilizing the endosomal sorting complex required for transport-I (ESCRT-I) component vacuolar protein sorting 23A (VPS23A) and thereby affect the stability of ABA receptors in Arabidopsis thaliana. Genetic analysis showed that VPS23A overexpression could rescue the ABA hypersensitive and drought tolerance phenotypes of ubp12-2w or ubp13-1. In addition to the direct regulation of VPS23A, we found that UBP12 and UBP13 also stabilized the E3 ligase XB3 ortholog 5 in A. thaliana (XBAT35.2) in response to ABA treatment. Hence, we demonstrated that UBP12 and UBP13 are previously unidentified rheostatic regulators of ABA signaling and revealed a mechanism by which deubiquitination precisely monitors the XBAT35/VPS23A ubiquitination module in the ABA response.

The soluble form of aluminum (Al) is a major constraint to crop production in acidic soils. The Al exclusion correlated with the Al-induced organic acid is considered as an important mechanism of Al resistance. The regulation of organic acid exudation in response to Al stress mediated by the root organic acid transporters has been extensively studied. However, how plants respond to Al stress through the regulation of organic acid homeostasis is not well understood. In this study, we identified the functionally unknown Transition zone1 (TZ1) as an Al-inducible gene in the root transition zone, the most sensitive region to Al stress, in Arabidopsis. tz1 mutants showed enhanced Al resistance and displayed greatly reduced root growth inhibition. Furthermore, TZ1 was found to interact with the aconitases (ACOs) which can catalyze the conversion from citrate, one of the most important organic acids, into isocitrate. Consistently, in tz1 mutants, the citric acid content was highly increased. Collectively, this study provides evidence to show that TZ1 negatively regulates root growth response to Al stress through interacting with ACOs and regulating citric acid homeostasis.