首页 > 最新文献

Plant signaling & behavior最新文献

英文 中文
Quasi-targeted metabolomics revealed isoliquiritigenin and lauric acid associated with resistance to tobacco black shank. 准靶向代谢组学揭示了与烟草黑柄病抗性相关的isisiquiritigenin和月桂酸。
Pub Date : 2024-12-31 Epub Date: 2024-03-25 DOI: 10.1080/15592324.2024.2332019
Shiwen Peng, Fangling Shu, Yanhui Lu, Dongsheng Fan, Dehong Zheng, Gaoqing Yuan

Tobacco black shank (TBS), caused by Phytophthora nicotianae, is a severe disease. Plant root exudates play a crucial role in mediating plant-pathogen interactions in the rhizosphere. However, the specific interaction between key secondary metabolites present in root exudates and the mechanisms of disease resistance remains poorly understood. This study conducted a comprehensive comparison via quasi-targeted metabolomic analysis on the root exudate metabolites from the tobacco cultivar Yunyan87 and K326, both before and after inoculation with P. nicotianae. The results showed that the root exudate metabolites changed after P. nicotianae inoculation, and the root exudate metabolites of different tobacco cultivar was significantly different. Furthermore, homovanillic acid, lauric acid, and isoliquiritigenin were identified as potential key compounds for TBS resistance based on their impact on the mycelium growth of the pathogens. The pot experiment showed that isoliquiritigenin reduced the incidence by 55.2%, while lauric acid reduced it by 45.8%. This suggests that isoliquiritigenin and lauric acid have potential applications in the management of TBS. In summary, this study revealed the possible resistance mechanisms of differential metabolites in resistance of commercial tobacco cultivar, and for the first time discovered the inhibitory effects of isoliquiritigenin and homovanillic acid on P. nictianae, and attempt to use plants secondary metabolites of for plant protection.

烟草黑柄病(TBS)是由烟草疫霉菌(Phytophthora nicotianae)引起的一种严重病害。植物根部渗出物在根圈中介导植物与病原体的相互作用方面发挥着至关重要的作用。然而,人们对根部渗出物中存在的关键次生代谢物与抗病机制之间的具体相互作用仍然知之甚少。本研究通过准靶向代谢组学分析,对烟草栽培品种云烟87和K326在接种烟粉虱前后的根部渗出物代谢物进行了全面比较。结果表明,烟粉虱接种后根部渗出物代谢物发生了变化,不同烟草品种的根部渗出物代谢物差异显著。此外,根据高香草酸、月桂酸和isoliquiritigenin对病原菌菌丝生长的影响,确定了它们是抗TBS的潜在关键化合物。盆栽实验表明,isoliquiritigenin 可使发病率降低 55.2%,而月桂酸可使发病率降低 45.8%。这表明,Iisiquiritigenin 和月桂酸有可能应用于 TBS 的管理。综上所述,本研究揭示了不同代谢物对商品烟草品种抗性的可能抗性机理,首次发现了isisiquiritigenin和homovanillic acid对烟粉虱的抑制作用,并尝试将植物次生代谢物用于植物保护。
{"title":"Quasi-targeted metabolomics revealed isoliquiritigenin and lauric acid associated with resistance to tobacco black shank.","authors":"Shiwen Peng, Fangling Shu, Yanhui Lu, Dongsheng Fan, Dehong Zheng, Gaoqing Yuan","doi":"10.1080/15592324.2024.2332019","DOIUrl":"10.1080/15592324.2024.2332019","url":null,"abstract":"<p><p>Tobacco black shank (TBS), caused by <i>Phytophthora nicotianae</i>, is a severe disease. Plant root exudates play a crucial role in mediating plant-pathogen interactions in the rhizosphere. However, the specific interaction between key secondary metabolites present in root exudates and the mechanisms of disease resistance remains poorly understood. This study conducted a comprehensive comparison via quasi-targeted metabolomic analysis on the root exudate metabolites from the tobacco cultivar Yunyan87 and K326, both before and after inoculation with <i>P. nicotianae</i>. The results showed that the root exudate metabolites changed after <i>P. nicotianae</i> inoculation, and the root exudate metabolites of different tobacco cultivar was significantly different. Furthermore, homovanillic acid, lauric acid, and isoliquiritigenin were identified as potential key compounds for TBS resistance based on their impact on the mycelium growth of the pathogens. The pot experiment showed that isoliquiritigenin reduced the incidence by 55.2%, while lauric acid reduced it by 45.8%. This suggests that isoliquiritigenin and lauric acid have potential applications in the management of TBS. In summary, this study revealed the possible resistance mechanisms of differential metabolites in resistance of commercial tobacco cultivar, and for the first time discovered the inhibitory effects of isoliquiritigenin and homovanillic acid on <i>P. nictianae</i>, and attempt to use plants secondary metabolites of for plant protection.</p>","PeriodicalId":94172,"journal":{"name":"Plant signaling & behavior","volume":"19 1","pages":"2332019"},"PeriodicalIF":0.0,"publicationDate":"2024-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10965109/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140290091","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
The 14-3-3 protein nt GF14e interacts with CIPK2 and increases low potassium stress in tobacco. 14-3-3 蛋白 nt GF14e 与 CIPK2 相互作用,增加烟草的低钾胁迫。
Pub Date : 2024-12-31 Epub Date: 2024-06-02 DOI: 10.1080/15592324.2024.2359257
Li Xu, Yifei Lu, Jiarui Jiang, Qian Chen, Yong Xu, Qili Mi, Haiying Xiang, Liming Lu, Xuemei Li, Qian Gao, Liqin Li

Potassium (K+) plays a role in enzyme activation, membrane transport, and osmotic regulation processes. An increase in potassium content can significantly improve the elasticity and combustibility of tobacco and reduce the content of harmful substances. Here, we report that the expression analysis of Nt GF14e, a 14-3-3 gene, increased markedly after low-potassium treatment (LK). Then, chlorophyll content, POD activity and potassium content, were significantly increased in overexpression of Nt GF14e transgenic tobacco lines compared with those in the wild type plants. The net K+ efflux rates were severely lower in the transgenic plants than in the wild type under LK stress. Furthermore, transcriptome analysis identified 5708 upregulated genes and 2787 downregulated genes between Nt GF14e overexpressing transgenic tobacco plants. The expression levels of some potassium-related genes were increased, such as CBL-interacting protein kinase 2 (CIPK2), Nt CIPK23, Nt CIPK25, H+-ATPase isoform 2 a (AHA2a), Nt AHA4a, Stelar K+ outward rectifier 1(SKOR1), and high affinity K+ transporter 5 (HAK5). The result of yeast two-hybrid and luciferase complementation imaging experiments suggested Nt GF14e could interact with CIPK2. Overall, these findings indicate that NtGF14e plays a vital roles in improving tobacco LK tolerance and enhancing potassium nutrition signaling pathways in tobacco plants.

钾(K+)在酶活化、膜运输和渗透调节过程中发挥作用。增加钾的含量可以明显改善烟草的弹性和可燃性,减少有害物质的含量。在此,我们报告了 14-3-3 基因 Nt GF14e 在低钾处理(LK)后的表达分析。与野生型植株相比,过表达 Nt GF14e 转基因烟草株系的叶绿素含量、POD 活性和钾含量均显著增加。在 LK 胁迫下,转基因植株的 K+ 净外流率明显低于野生型植株。此外,转录组分析还发现了 Nt GF14e 过表达转基因烟草植株的 5708 个上调基因和 2787 个下调基因。一些与钾有关的基因表达水平升高,如 CBL-互作蛋白激酶 2(CIPK2)、Nt CIPK23、Nt CIPK25、H+-ATPase isoform 2 a(AHA2a)、Nt AHA4a、Stelar K+ outward rectifier 1(SKOR1)和高亲和性 K+ 转运体 5(HAK5)。酵母双杂交和荧光素酶互补成像实验的结果表明,Nt GF14e能与CIPK2相互作用。总之,这些研究结果表明,NtGF14e在提高烟草耐钾性和增强烟草植物钾营养信号通路中发挥着重要作用。
{"title":"The 14-3-3 protein nt GF14e interacts with CIPK2 and increases low potassium stress in tobacco.","authors":"Li Xu, Yifei Lu, Jiarui Jiang, Qian Chen, Yong Xu, Qili Mi, Haiying Xiang, Liming Lu, Xuemei Li, Qian Gao, Liqin Li","doi":"10.1080/15592324.2024.2359257","DOIUrl":"10.1080/15592324.2024.2359257","url":null,"abstract":"<p><p>Potassium (K<sup>+</sup>) plays a role in enzyme activation, membrane transport, and osmotic regulation processes. An increase in potassium content can significantly improve the elasticity and combustibility of tobacco and reduce the content of harmful substances. Here, we report that the expression analysis of <i>Nt GF14e</i>, a <i>14-3-3</i> gene, increased markedly after low-potassium treatment (LK). Then, chlorophyll content, POD activity and potassium content, were significantly increased in overexpression of <i>Nt GF14e</i> transgenic tobacco lines compared with those in the wild type plants. The net K<sup>+</sup> efflux rates were severely lower in the transgenic plants than in the wild type under LK stress. Furthermore, transcriptome analysis identified 5708 upregulated genes and 2787 downregulated genes between <i>Nt GF14e</i> overexpressing transgenic tobacco plants. The expression levels of some potassium-related genes were increased, such as CBL-interacting protein kinase 2 (<i>CIPK2</i>), <i>Nt CIPK23</i>, <i>Nt CIPK25</i>, H<sup>+</sup>-ATPase isoform 2 <i>a</i> (<i>AHA2a</i>), <i>Nt AHA4a</i>, Stelar K<sup>+</sup> outward rectifier 1(<i>SKOR1</i>), and high affinity K<sup>+</sup> transporter 5 (<i>HAK5</i>). The result of yeast two-hybrid and luciferase complementation imaging experiments suggested Nt GF14e could interact with CIPK2. Overall, these findings indicate that NtGF14e plays a vital roles in improving tobacco LK tolerance and enhancing potassium nutrition signaling pathways in tobacco plants.</p>","PeriodicalId":94172,"journal":{"name":"Plant signaling & behavior","volume":"19 1","pages":"2359257"},"PeriodicalIF":0.0,"publicationDate":"2024-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11152103/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141201339","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Identification of mitogen-activated protein kinases substrates in Arabidopsis using kinase client assay. 利用激酶客户测定法鉴定拟南芥中的丝裂原活化蛋白激酶底物。
Pub Date : 2024-12-31 Epub Date: 2024-03-17 DOI: 10.1080/15592324.2024.2326238
Sunghwa Bahk, Nagib Ahsan, Jonguk An, Sun Ho Kim, Zakiyah Ramadany, Jong Chan Hong, Jay J Thelen, Woo Sik Chung

Mitogen-activated protein kinase (MPK) cascades are essential signal transduction components that control a variety of cellular responses in all eukaryotes. MPKs convert extracellular stimuli into cellular responses by the phosphorylation of downstream substrates. Although MPK cascades are predicted to be very complex, only limited numbers of MPK substrates have been identified in plants. Here, we used the kinase client (KiC) assay to identify novel substrates of MPK3 and MPK6. Recombinant MPK3 or MPK6 were tested against a large synthetic peptide library representing in vivo phosphorylation sites, and phosphorylated peptides were identified by high-resolution tandem mass spectrometry. From this screen, we identified 23 and 21 putative client peptides of MPK3 and MPK6, respectively. To verify the phosphorylation of putative client peptides, we performed in vitro kinase assay with recombinant fusion proteins of isolated client peptides. We found that 13 and 9 recombinant proteins were phosphorylated by MPK3 and MPK6. Among them, 11 proteins were proven to be the novel substrates of two MPKs. This study suggests that the KiC assay is a useful method to identify new substrates of MPKs.

丝裂原活化蛋白激酶(MPK)级联是控制所有真核生物各种细胞反应的重要信号转导元件。MPK 通过下游底物的磷酸化将细胞外刺激转化为细胞反应。虽然 MPK 级联被认为非常复杂,但在植物中只发现了有限数量的 MPK 底物。在这里,我们使用激酶客户(KiC)测定法来鉴定 MPK3 和 MPK6 的新型底物。用代表体内磷酸化位点的大型合成肽库对重组 MPK3 或 MPK6 进行测试,并通过高分辨率串联质谱鉴定磷酸化肽。通过这一筛选,我们分别鉴定出了 23 个和 21 个 MPK3 和 MPK6 的假定客户肽。为了验证推定客户肽的磷酸化情况,我们用分离出的客户肽的重组融合蛋白进行了体外激酶试验。我们发现,分别有 13 个和 9 个重组蛋白被 MPK3 和 MPK6 磷酸化。其中,11 个蛋白被证明是两种 MPK 的新型底物。这项研究表明,KiC测定是一种鉴别MPK新底物的有效方法。
{"title":"Identification of mitogen-activated protein kinases substrates in <i>Arabidopsis</i> using kinase client assay.","authors":"Sunghwa Bahk, Nagib Ahsan, Jonguk An, Sun Ho Kim, Zakiyah Ramadany, Jong Chan Hong, Jay J Thelen, Woo Sik Chung","doi":"10.1080/15592324.2024.2326238","DOIUrl":"10.1080/15592324.2024.2326238","url":null,"abstract":"<p><p>Mitogen-activated protein kinase (MPK) cascades are essential signal transduction components that control a variety of cellular responses in all eukaryotes. MPKs convert extracellular stimuli into cellular responses by the phosphorylation of downstream substrates. Although MPK cascades are predicted to be very complex, only limited numbers of MPK substrates have been identified in plants. Here, we used the kinase client (KiC) assay to identify novel substrates of MPK3 and MPK6. Recombinant MPK3 or MPK6 were tested against a large synthetic peptide library representing <i>in vivo</i> phosphorylation sites, and phosphorylated peptides were identified by high-resolution tandem mass spectrometry. From this screen, we identified 23 and 21 putative client peptides of MPK3 and MPK6, respectively. To verify the phosphorylation of putative client peptides, we performed <i>in vitro</i> kinase assay with recombinant fusion proteins of isolated client peptides. We found that 13 and 9 recombinant proteins were phosphorylated by MPK3 and MPK6. Among them, 11 proteins were proven to be the novel substrates of two MPKs. This study suggests that the KiC assay is a useful method to identify new substrates of MPKs.</p>","PeriodicalId":94172,"journal":{"name":"Plant signaling & behavior","volume":"19 1","pages":"2326238"},"PeriodicalIF":0.0,"publicationDate":"2024-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10950278/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140144907","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Novel perspectives on plant behavior. 植物行为的新视角
Pub Date : 2024-12-31 Epub Date: 2024-11-04 DOI: 10.1080/15592324.2024.2419673
E Van Volkenburgh, E D Brenner
{"title":"Novel perspectives on plant behavior.","authors":"E Van Volkenburgh, E D Brenner","doi":"10.1080/15592324.2024.2419673","DOIUrl":"10.1080/15592324.2024.2419673","url":null,"abstract":"","PeriodicalId":94172,"journal":{"name":"Plant signaling & behavior","volume":"19 1","pages":"2419673"},"PeriodicalIF":0.0,"publicationDate":"2024-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11540076/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142577339","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Exploring cotton SFR2's conundrum in response to cold stress. 探索棉花 SFR2 在应对寒冷胁迫时的难题。
Pub Date : 2024-12-31 Epub Date: 2024-06-05 DOI: 10.1080/15592324.2024.2362518
Samantha M Surber, Ngoc Pham Thien Thao, Cailin N Smith, Zachery D Shomo, Allison C Barnes, Rebecca L Roston

Cotton is an important agricultural crop to many regions across the globe but is sensitive to low-temperature exposure. The activity of the enzyme SENSITIVE TO FREEZING 2 (SFR2) improves cold tolerance of plants and produces trigalactosylsyldiacylglycerol (TGDG), but its role in cold sensitive plants, such as cotton remains unknown. Recently, it was reported that cotton SFR2 produced very little TGDG under normal and cold conditions. Here, we investigate cotton SFR2 activation and TGDG production. Using multiple approaches in the native system and transformation into Arabidopsis thaliana, as well as heterologous yeast expression, we provide evidence that cotton SFR2 activates differently than previously found among other plant species. We conclude with the hypothesis that SFR2 in cotton is not activated in a similar manner regarding acidification or freezing like Arabidopsis and that other regions of SFR2 protein are critical for activation of the enzyme than previously reported.

棉花是全球许多地区的重要农作物,但对低温很敏感。对冷冻敏感的酶 2(SFR2)的活性提高了植物的耐寒性,并产生了三聚半乳糖基酰二乙酰甘油(TGDG),但它在对冷敏感的植物(如棉花)中的作用仍然未知。最近有报道称,棉花 SFR2 在正常和寒冷条件下产生的 TGDG 都很少。在此,我们研究了棉花 SFR2 的活化和 TGDG 的产生。通过在原生系统中使用多种方法、转化到拟南芥中以及异源酵母表达,我们提供了棉花 SFR2 的活化方式与之前在其他植物物种中发现的不同的证据。我们的结论是,棉花中的 SFR2 并不像拟南芥那样在酸化或冷冻时以类似的方式激活,而且 SFR2 蛋白的其他区域对于酶的激活比以前报道的更为关键。
{"title":"Exploring cotton SFR2's conundrum in response to cold stress.","authors":"Samantha M Surber, Ngoc Pham Thien Thao, Cailin N Smith, Zachery D Shomo, Allison C Barnes, Rebecca L Roston","doi":"10.1080/15592324.2024.2362518","DOIUrl":"10.1080/15592324.2024.2362518","url":null,"abstract":"<p><p>Cotton is an important agricultural crop to many regions across the globe but is sensitive to low-temperature exposure. The activity of the enzyme SENSITIVE TO FREEZING 2 (SFR2) improves cold tolerance of plants and produces trigalactosylsyldiacylglycerol (TGDG), but its role in cold sensitive plants, such as cotton remains unknown. Recently, it was reported that cotton SFR2 produced very little TGDG under normal and cold conditions. Here, we investigate cotton SFR2 activation and TGDG production. Using multiple approaches in the native system and transformation into <i>Arabidopsis thaliana</i>, as well as heterologous yeast expression, we provide evidence that cotton SFR2 activates differently than previously found among other plant species. We conclude with the hypothesis that SFR2 in cotton is not activated in a similar manner regarding acidification or freezing like Arabidopsis and that other regions of SFR2 protein are critical for activation of the enzyme than previously reported.</p>","PeriodicalId":94172,"journal":{"name":"Plant signaling & behavior","volume":"19 1","pages":"2362518"},"PeriodicalIF":0.0,"publicationDate":"2024-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11155703/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141249272","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Physiological and transcriptomic analysis of tomato in response to sub-optimal temperature stress. 番茄对次优温度胁迫的生理和转录组分析。
Pub Date : 2024-12-31 Epub Date: 2024-03-21 DOI: 10.1080/15592324.2024.2332018
Huan Gao, Fengzhi Wu

Tomato (Solanum lycopersicum L.) is one of the most important economic crops in China. However, its quality and yield are susceptible to the adverse effects of low temperatures. In our study, two tomato cultivars, showing different tolerance to low temperatures, namely the cold-sensitive tomato cultivar (S708) and cold-tolerant tomato cultivar (T722), were grown at optimal (25/18°C) and sub-optimal (15/10°C) temperature conditions for 5 days. Our study aimed to explore the effect of sub-optimal temperature on fresh weight, chlorophyll content and chlorophyll fluorescence, soluble sugars and proline content of two tomato cultivars. Moreover, we employed RNA-Seq to analyze the transcriptomic response of tomato roots to sub-optimal temperature. The results revealed that S708 showed a more significant reduction in fresh weight, chlorophyll content, photochemical efficiency of PSII (YII), maximum quantum yield of PSII (Fv/Fm), photochemical quenching (qP) and electron transport rate (ETR) compared to T722 under the sub-optimal temperature condition. Notably, T722 maintained higher level of soluble sugars and proline in comparison to S708 uner sub-optimal temperature. RNA-seq data showed that up-regulated DEGs in both tomato cultivars were involved in "plant-pathogen interaction", "MAPK signaling pathway", "plant hormone signal transduction", and "phosphatidylinositol signaling system". Furthermore, "Amino sugar and nucleotide sugar metabolism" pathway was enriched only in T722. Moreover, under sub-optimal temperature, transcription factor genes and osmoregulation genes showed varying degrees of response in both tomato cultivars. Conclusion: In summary, our results offer detailed insights into the response characteristics of tomato to sub-optimal temperature, providing valuable references for the practical management of tomato crops under sub-optimal temperature condition.

番茄(Solanum lycopersicum L.)是中国最重要的经济作物之一。然而,其品质和产量易受低温的不利影响。在我们的研究中,两个对低温有不同耐受性的番茄栽培品种,即冷敏感番茄栽培品种(S708)和冷耐受番茄栽培品种(T722),分别在最适温度(25/18°C)和次优温度(15/10°C)条件下生长了 5 天。我们的研究旨在探讨次优温度对两个番茄品种的鲜重、叶绿素含量和叶绿素荧光、可溶性糖和脯氨酸含量的影响。此外,我们还利用 RNA-Seq 分析了番茄根系对次优温度的转录组响应。结果表明,在次优温度条件下,与 T722 相比,S708 的鲜重、叶绿素含量、PSII 的光化学效率(YII)、PSII 的最大量子产率(Fv/Fm)、光化学淬灭(qP)和电子传输速率(ETR)都有更显著的下降。值得注意的是,与 S708 相比,T722 在次优温度条件下保持了更高的可溶性糖和脯氨酸水平。RNA-seq 数据显示,两个番茄品种中上调的 DEGs 涉及 "植物与病原体相互作用"、"MAPK 信号通路"、"植物激素信号转导 "和 "磷脂酰肌醇信号系统"。此外,"氨基酸糖和核苷酸糖代谢 "途径仅在 T722 中富集。此外,在次优温度条件下,两个番茄品种的转录因子基因和渗透调节基因均表现出不同程度的反应。结论总之,我们的研究结果详细揭示了番茄对次优温度的响应特征,为番茄作物在次优温度条件下的实际管理提供了有价值的参考。
{"title":"Physiological and transcriptomic analysis of tomato in response to sub-optimal temperature stress.","authors":"Huan Gao, Fengzhi Wu","doi":"10.1080/15592324.2024.2332018","DOIUrl":"10.1080/15592324.2024.2332018","url":null,"abstract":"<p><p>Tomato (<i>Solanum lycopersicum</i> L.) is one of the most important economic crops in China. However, its quality and yield are susceptible to the adverse effects of low temperatures. In our study, two tomato cultivars, showing different tolerance to low temperatures, namely the cold-sensitive tomato cultivar (S708) and cold-tolerant tomato cultivar (T722), were grown at optimal (25/18°C) and sub-optimal (15/10°C) temperature conditions for 5 days. Our study aimed to explore the effect of sub-optimal temperature on fresh weight, chlorophyll content and chlorophyll fluorescence, soluble sugars and proline content of two tomato cultivars. Moreover, we employed RNA-Seq to analyze the transcriptomic response of tomato roots to sub-optimal temperature. The results revealed that S708 showed a more significant reduction in fresh weight, chlorophyll content, photochemical efficiency of PSII (YII), maximum quantum yield of PSII (Fv/Fm), photochemical quenching (qP) and electron transport rate (ETR) compared to T722 under the sub-optimal temperature condition. Notably, T722 maintained higher level of soluble sugars and proline in comparison to S708 uner sub-optimal temperature. RNA-seq data showed that up-regulated DEGs in both tomato cultivars were involved in \"plant-pathogen interaction\", \"MAPK signaling pathway\", \"plant hormone signal transduction\", and \"phosphatidylinositol signaling system\". Furthermore, \"Amino sugar and nucleotide sugar metabolism\" pathway was enriched only in T722. Moreover, under sub-optimal temperature, transcription factor genes and osmoregulation genes showed varying degrees of response in both tomato cultivars. Conclusion: In summary, our results offer detailed insights into the response characteristics of tomato to sub-optimal temperature, providing valuable references for the practical management of tomato crops under sub-optimal temperature condition.</p>","PeriodicalId":94172,"journal":{"name":"Plant signaling & behavior","volume":"19 1","pages":"2332018"},"PeriodicalIF":0.0,"publicationDate":"2024-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10962623/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140178419","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Luciferase-mediated assay to detect the PAMP-triggered gene expression in transgenic Nicotiana benthamiana. 以荧光素酶为介导的检测方法,用于检测转基因烟草中 PAMP 触发的基因表达。
Pub Date : 2024-12-31 Epub Date: 2024-11-25 DOI: 10.1080/15592324.2024.2411918
Ola Barakat, Fushuang Zhang, Mengzhu Zeng, Yuanchao Wang

Luciferase is one of the bioluminescence-producing agents, which was widely used as a reporter enzyme for constructing bioassay systems to study gene expression with high accuracy and within a broad dynamic spectrum. Perception of pathogen associated molecular patterns (PAMPs) in plants often lead to significant transcriptional changes. The transcriptional changes of defense-related genes are often used as a marker to assay PAMP-triggered plant immune response. In this study, we showed that the marker gene CYP71D20 was rapidly activated in Nicotiana benthamiana upon treatment with the bacterial PAMP flg22 and the Phytophthora elicitin INF1. In addition, we generated transgenic N. benthamiana using the luciferase as a reporter gene to analyze CYP71D20 gene expression upon PAMP treatment. The transgenic line carrying the luciferase gene driven by CYP71D20 promoter was treated with the bacterial PAMP flg22 or Phytophthora elicitin INF1. Transcriptional activation of CYP71D20 was measured by monitoring the luciferase activity. The results showed that the LUC activity was increased after treatment with different PAMPs, indicating that the CYP71D20-promotor luciferase assay can be used to study the PAMP-triggered gene expression in N. benthamiana.

荧光素酶是生物发光剂之一,被广泛用作构建生物测定系统的报告酶,以高精度和宽动态光谱研究基因表达。植物对病原体相关分子模式(PAMPs)的感知往往会导致显著的转录变化。防御相关基因的转录变化通常被用作检测 PAMP 触发的植物免疫反应的标记。在本研究中,我们发现在用细菌 PAMP flg22 和 Phytophthora elicitin INF1 处理烟草后,标记基因 CYP71D20 被迅速激活。此外,我们还利用荧光素酶作为报告基因生成了转基因烟草,以分析 PAMP 处理时 CYP71D20 基因的表达情况。将携带由 CYP71D20 启动子驱动的荧光素酶基因的转基因品系用细菌 PAMP flg22 或 Phytophthora elicitin INF1 处理。通过监测荧光素酶活性来测量 CYP71D20 的转录激活情况。结果表明,经不同的 PAMP 处理后,荧光素酶活性均有所提高,这表明 CYP71D20 启动子荧光素酶检测法可用于研究 PAMP 触发的 N. benthamiana 基因表达。
{"title":"Luciferase-mediated assay to detect the PAMP-triggered gene expression in transgenic <i>Nicotiana benthamiana</i>.","authors":"Ola Barakat, Fushuang Zhang, Mengzhu Zeng, Yuanchao Wang","doi":"10.1080/15592324.2024.2411918","DOIUrl":"10.1080/15592324.2024.2411918","url":null,"abstract":"<p><p>Luciferase is one of the bioluminescence-producing agents, which was widely used as a reporter enzyme for constructing bioassay systems to study gene expression with high accuracy and within a broad dynamic spectrum. Perception of pathogen associated molecular patterns (PAMPs) in plants often lead to significant transcriptional changes. The transcriptional changes of defense-related genes are often used as a marker to assay PAMP-triggered plant immune response. In this study, we showed that the marker gene <i>CYP71D20</i> was rapidly activated in <i>Nicotiana benthamiana</i> upon treatment with the bacterial PAMP flg22 and the Phytophthora elicitin INF1. In addition, we generated transgenic <i>N. benthamiana</i> using the luciferase as a reporter gene to analyze <i>CYP71D20</i> gene expression upon PAMP treatment. The transgenic line carrying the luciferase gene driven by <i>CYP71D20</i> promoter was treated with the bacterial PAMP flg22 or Phytophthora elicitin INF1. Transcriptional activation of <i>CYP71D20</i> was measured by monitoring the luciferase activity. The results showed that the LUC activity was increased after treatment with different PAMPs, indicating that the <i>CYP71D20</i>-promotor luciferase assay can be used to study the PAMP-triggered gene expression in <i>N. benthamiana</i>.</p>","PeriodicalId":94172,"journal":{"name":"Plant signaling & behavior","volume":"19 1","pages":"2411918"},"PeriodicalIF":0.0,"publicationDate":"2024-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11591477/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142711747","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Plant ultrasound detection: a cost-effective method for identifying plant ultrasonic emissions. 植物超声波探测:一种识别植物超声波发射的经济有效的方法。
Pub Date : 2024-12-31 Epub Date: 2024-02-12 DOI: 10.1080/15592324.2024.2310974
Luca Bonisoli, Ivano Pelicella, Laura Arru

Plants have been observed to produce short ultrasonic emissions (UEs), and current research is focusing on developing noninvasive techniques for recording and analyzing these emissions. A standardized methodology has not been established yet; in this paper we suggest a cost-effective procedure for recording, extracting, and identifying plant UEs using only a single ultrasound microphone, a laptop computer, and open-source software.

据观察,植物会产生短促的超声波发射(UEs),目前的研究重点是开发记录和分析这些发射的非侵入式技术。本文提出了一种经济有效的方法,只需使用一个超声波麦克风、一台笔记本电脑和开源软件,就能记录、提取和识别植物的超声波发射。
{"title":"Plant ultrasound detection: a cost-effective method for identifying plant ultrasonic emissions.","authors":"Luca Bonisoli, Ivano Pelicella, Laura Arru","doi":"10.1080/15592324.2024.2310974","DOIUrl":"10.1080/15592324.2024.2310974","url":null,"abstract":"<p><p>Plants have been observed to produce short ultrasonic emissions (UEs), and current research is focusing on developing noninvasive techniques for recording and analyzing these emissions. A standardized methodology has not been established yet; in this paper we suggest a cost-effective procedure for recording, extracting, and identifying plant UEs using only a single ultrasound microphone, a laptop computer, and open-source software.</p>","PeriodicalId":94172,"journal":{"name":"Plant signaling & behavior","volume":"19 1","pages":"2310974"},"PeriodicalIF":0.0,"publicationDate":"2024-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10863351/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139725523","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Proteome-wide analysis reveals G protein-coupled receptor-like proteins in rice (Oryza sativa). 全蛋白质组分析揭示了水稻(Oryza sativa)中的 G 蛋白偶联受体样蛋白。
Pub Date : 2024-12-31 Epub Date: 2024-06-21 DOI: 10.1080/15592324.2024.2365572
Dinesh K Yadav, Gyan Prakash Srivastava, Ananya Singh, Madhavi Singh, Neelam Yadav, Narendra Tuteja

G protein-coupled receptors (GPCRs) constitute the largest family of transmembrane proteins in metazoans that mediate the regulation of various physiological responses to discrete ligands through heterotrimeric G protein subunits. The existence of GPCRs in plant is contentious, but their comparable crucial role in various signaling pathways necessitates the identification of novel remote GPCR-like proteins that essentially interact with the plant G protein α subunit and facilitate the transduction of various stimuli. In this study, we identified three putative GPCR-like proteins (OsGPCRLPs) (LOC_Os06g09930.1, LOC_Os04g36630.1, and LOC_Os01g54784.1) in the rice proteome using a stringent bioinformatics workflow. The identified OsGPCRLPs exhibited a canonical GPCR 'type I' 7TM topology, patterns, and biologically significant sites for membrane anchorage and desensitization. Cluster-based interactome mapping revealed that the identified proteins interact with the G protein α subunit which is a characteristic feature of GPCRs. Computational results showing the interaction of identified GPCR-like proteins with G protein α subunit and its further validation by the membrane yeast-two-hybrid assay strongly suggest the presence of GPCR-like 7TM proteins in the rice proteome. The absence of a regulator of G protein signaling (RGS) box in the C- terminal domain, and the presence of signature motifs of canonical GPCR in the identified OsGPCRLPs strongly suggest that the rice proteome contains GPCR-like proteins that might be involved in signal transduction.

G 蛋白偶联受体(GPCR)是元类动物中最大的跨膜蛋白家族,通过异三聚体 G 蛋白亚基介导对离散配体的各种生理反应的调节。植物中是否存在 GPCR 尚存争议,但由于它们在各种信号通路中发挥着相当关键的作用,因此有必要鉴定新型的远端 GPCR 类蛋白,这些蛋白本质上与植物 G 蛋白 α 亚基相互作用,促进各种刺激的转导。在本研究中,我们采用严格的生物信息学工作流程,在水稻蛋白质组中鉴定出了三个假定的类 GPCR 蛋白(OsGPCRLPs)(LOC_Os06g09930.1、LOC_Os04g36630.1 和 LOC_Os01g54784.1)。鉴定出的 OsGPCRLPs 具有典型的 GPCR "I 型 "7TM 拓扑结构、模式以及具有生物学意义的膜锚定和脱敏位点。基于聚类的相互作用组图谱显示,鉴定出的蛋白质与 G 蛋白 α 亚基相互作用,这是 GPCR 的一个特征。计算结果显示了已鉴定的类 GPCR 蛋白与 G 蛋白 α 亚基的相互作用,并通过膜酵母-双杂交试验进一步验证了这一点,这有力地表明了水稻蛋白质组中存在类 GPCR 7TM 蛋白。在已鉴定的 OsGPCRLPs 的 C 端结构域中没有 G 蛋白信号转导调节器(RGS)框,而且存在典型 GPCR 的标志性基序,这些都有力地表明水稻蛋白质组中含有可能参与信号转导的 GPCR 样蛋白。
{"title":"Proteome-wide analysis reveals G protein-coupled receptor-like proteins in rice (<i>Oryza sativa</i>).","authors":"Dinesh K Yadav, Gyan Prakash Srivastava, Ananya Singh, Madhavi Singh, Neelam Yadav, Narendra Tuteja","doi":"10.1080/15592324.2024.2365572","DOIUrl":"10.1080/15592324.2024.2365572","url":null,"abstract":"<p><p>G protein-coupled receptors (GPCRs) constitute the largest family of transmembrane proteins in metazoans that mediate the regulation of various physiological responses to discrete ligands through heterotrimeric G protein subunits. The existence of GPCRs in plant is contentious, but their comparable crucial role in various signaling pathways necessitates the identification of novel remote GPCR-like proteins that essentially interact with the plant G protein α subunit and facilitate the transduction of various stimuli. In this study, we identified three putative GPCR-like proteins (OsGPCRLPs) (LOC_Os06g09930.1, LOC_Os04g36630.1, and LOC_Os01g54784.1) in the rice proteome using a stringent bioinformatics workflow. The identified OsGPCRLPs exhibited a canonical GPCR 'type I' 7TM topology, patterns, and biologically significant sites for membrane anchorage and desensitization. Cluster-based interactome mapping revealed that the identified proteins interact with the G protein α subunit which is a characteristic feature of GPCRs. Computational results showing the interaction of identified GPCR-like proteins with G protein α subunit and its further validation by the membrane yeast-two-hybrid assay strongly suggest the presence of GPCR-like 7TM proteins in the rice proteome. The absence of a regulator of G protein signaling (RGS) box in the C- terminal domain, and the presence of signature motifs of canonical GPCR in the identified OsGPCRLPs strongly suggest that the rice proteome contains GPCR-like proteins that might be involved in signal transduction.</p>","PeriodicalId":94172,"journal":{"name":"Plant signaling & behavior","volume":"19 1","pages":"2365572"},"PeriodicalIF":0.0,"publicationDate":"2024-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11195488/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141433737","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Nitric oxide in plants: an insight on redox activity and responses toward abiotic stress signaling. 植物中的一氧化氮:对氧化还原活动和非生物胁迫信号反应的认识。
Pub Date : 2024-12-31 Epub Date: 2024-01-08 DOI: 10.1080/15592324.2023.2298053
Khushboo Khator, Suman Parihar, Jan Jasik, Gyan Singh Shekhawat

Plants, as sessile organisms, are subjected to diverse abiotic stresses, including salinity, desiccation, metal toxicity, thermal fluctuations, and hypoxia at different phases of plant growth. Plants can activate messenger molecules to initiate a signaling cascade of response toward environmental stresses that results in either cell death or plant acclimation. Nitric oxide (NO) is a small gaseous redox-active molecule that exhibits a plethora of physiological functions in growth, development, flowering, senescence, stomata closure and responses to environmental stresses. It can also facilitate alteration in protein function and reprogram the gene profiling by direct or indirect interaction with different target molecules. The bioactivity of NO can be manifested through different redox-based protein modifications including S-nitrosylation, protein nitration, and metal nitrosylation in plants. Although there has been considerable progress in the role of NO in regulating stress signaling, still the physiological mechanisms regarding the abiotic stress tolerance in plants remain unclear. This review summarizes recent advances in understanding the emerging knowledge regarding NO function in plant tolerance against abiotic stresses. The manuscript also highlighted the importance of NO as an abiotic stress modulator and developed a rational design for crop cultivation under a stress environment.

植物作为无梗生物,在其生长的不同阶段会受到各种非生物胁迫,包括盐度、干燥、金属毒性、热波动和缺氧。植物可以激活信使分子,启动信号级联反应,以应对环境胁迫,从而导致细胞死亡或植物适应环境。一氧化氮(NO)是一种具有氧化还原作用的气态小分子,在生长、发育、开花、衰老、气孔关闭和对环境胁迫的反应中具有多种生理功能。它还能通过与不同靶分子的直接或间接相互作用,促进蛋白质功能的改变和基因谱的重编程。氮氧化物的生物活性可通过不同的氧化还原型蛋白质修饰来体现,包括植物体内的 S-亚硝基化、蛋白质硝化和金属亚硝基化。尽管在氮氧化物调节胁迫信号转导的作用方面取得了很大进展,但植物耐受非生物胁迫的生理机制仍不清楚。这篇综述总结了在了解有关 NO 在植物耐受非生物胁迫中的功能的新知识方面的最新进展。稿件还强调了 NO 作为一种非生物胁迫调节剂的重要性,并对胁迫环境下的作物栽培进行了合理设计。
{"title":"Nitric oxide in plants: an insight on redox activity and responses toward abiotic stress signaling.","authors":"Khushboo Khator, Suman Parihar, Jan Jasik, Gyan Singh Shekhawat","doi":"10.1080/15592324.2023.2298053","DOIUrl":"10.1080/15592324.2023.2298053","url":null,"abstract":"<p><p>Plants, as sessile organisms, are subjected to diverse abiotic stresses, including salinity, desiccation, metal toxicity, thermal fluctuations, and hypoxia at different phases of plant growth. Plants can activate messenger molecules to initiate a signaling cascade of response toward environmental stresses that results in either cell death or plant acclimation. Nitric oxide (NO) is a small gaseous redox-active molecule that exhibits a plethora of physiological functions in growth, development, flowering, senescence, stomata closure and responses to environmental stresses. It can also facilitate alteration in protein function and reprogram the gene profiling by direct or indirect interaction with different target molecules. The bioactivity of NO can be manifested through different redox-based protein modifications including <i>S</i>-nitrosylation, protein nitration, and metal nitrosylation in plants. Although there has been considerable progress in the role of NO in regulating stress signaling, still the physiological mechanisms regarding the abiotic stress tolerance in plants remain unclear. This review summarizes recent advances in understanding the emerging knowledge regarding NO function in plant tolerance against abiotic stresses. The manuscript also highlighted the importance of NO as an abiotic stress modulator and developed a rational design for crop cultivation under a stress environment.</p>","PeriodicalId":94172,"journal":{"name":"Plant signaling & behavior","volume":"19 1","pages":"2298053"},"PeriodicalIF":0.0,"publicationDate":"2024-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10793691/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139405809","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
期刊
Plant signaling & behavior
全部 Acc. Chem. Res. ACS Applied Bio Materials ACS Appl. Electron. Mater. ACS Appl. Energy Mater. ACS Appl. Mater. Interfaces ACS Appl. Nano Mater. ACS Appl. Polym. Mater. ACS BIOMATER-SCI ENG ACS Catal. ACS Cent. Sci. ACS Chem. Biol. ACS Chemical Health & Safety ACS Chem. Neurosci. ACS Comb. Sci. ACS Earth Space Chem. ACS Energy Lett. ACS Infect. Dis. ACS Macro Lett. ACS Mater. Lett. ACS Med. Chem. Lett. ACS Nano ACS Omega ACS Photonics ACS Sens. ACS Sustainable Chem. Eng. ACS Synth. Biol. Anal. Chem. BIOCHEMISTRY-US Bioconjugate Chem. BIOMACROMOLECULES Chem. Res. Toxicol. Chem. Rev. Chem. Mater. CRYST GROWTH DES ENERG FUEL Environ. Sci. Technol. Environ. Sci. Technol. Lett. Eur. J. Inorg. Chem. IND ENG CHEM RES Inorg. Chem. J. Agric. Food. Chem. J. Chem. Eng. Data J. Chem. Educ. J. Chem. Inf. Model. J. Chem. Theory Comput. J. Med. Chem. J. Nat. Prod. J PROTEOME RES J. Am. Chem. Soc. LANGMUIR MACROMOLECULES Mol. Pharmaceutics Nano Lett. Org. Lett. ORG PROCESS RES DEV ORGANOMETALLICS J. Org. Chem. J. Phys. Chem. J. Phys. Chem. A J. Phys. Chem. B J. Phys. Chem. C J. Phys. Chem. Lett. Analyst Anal. Methods Biomater. Sci. Catal. Sci. Technol. Chem. Commun. Chem. Soc. Rev. CHEM EDUC RES PRACT CRYSTENGCOMM Dalton Trans. Energy Environ. Sci. ENVIRON SCI-NANO ENVIRON SCI-PROC IMP ENVIRON SCI-WAT RES Faraday Discuss. Food Funct. Green Chem. Inorg. Chem. Front. Integr. Biol. J. Anal. At. Spectrom. J. Mater. Chem. A J. Mater. Chem. B J. Mater. Chem. C Lab Chip Mater. Chem. Front. Mater. Horiz. MEDCHEMCOMM Metallomics Mol. Biosyst. Mol. Syst. Des. Eng. Nanoscale Nanoscale Horiz. Nat. Prod. Rep. New J. Chem. Org. Biomol. Chem. Org. Chem. Front. PHOTOCH PHOTOBIO SCI PCCP Polym. Chem.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1