生命科学最新文献

{"title":"Common gardens reveal genomic susceptibility and vulnerability to climate change in Eucalyptus","authors":"Xianliang Zhu,&nbsp;Hairun Zhang,&nbsp;Zhaohua Lu,&nbsp;Ming Kang,&nbsp;Baosheng Wang,&nbsp;David Bush,&nbsp;Changrong Li,&nbsp;Fagen Li","doi":"10.1111/tpj.70336","DOIUrl":"https://doi.org/10.1111/tpj.70336","url":null,"abstract":"<div>\u0000 \u0000 <p>Accelerated global climate change and increased species introduction across international scales have raised concerns about the potential for trees to experience maladaptation or lagging adaptation in response to these environmental shifts. However, our knowledge regarding the relationship between the genomic metrics used to predict maladaptation and actual fitness proxies in trees remains limited. Here, we present a population genomic analysis of 295 families from 28 provenances of <i>Eucalyptus pellita</i>, a widely cultivated fast-growing tree species, and conducted two common garden experiments. Genomic susceptibility encompassing individual heterozygosity (<i>H</i>), genomic inbreeding (<i>F</i>_ROH), and genomic load (inferred from deleterious mutations) exhibited distinct geographic patterns, shedding light on the origin and evolutionary history of <i>E. pellita</i>. The genetic basis of local adaptation was elucidated through genotype–environment associations and genome-wide association studies, including 198 loci associated with climate and 2388 loci regulating different traits. Furthermore, Australian provenances have higher genomic vulnerability under prospective climate alterations than Papua New Guinea and Indonesia provenances. By integrating phenotypic data across two common gardens, the relationship between leaf functional traits and predicted metrics of maladaptation was closer than growth attributes. Notably, pronounced natural selection signals linked to leaf morphogenesis have been identified by comparing two lineages spanning the oceans. This study underscores the immense potential of leveraging genomic susceptibility and genomic vulnerability to decipher the local (mal)adaptation of forest trees.</p>\u0000 </div>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"123 1","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-07-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144581960","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Correction to \"Verticillium dahliae Elicitor VdSP8 Enhances Disease Resistance Through Increasing Lignin Biosynthesis in Cotton\".","authors":"","doi":"10.1111/pce.70068","DOIUrl":"https://doi.org/10.1111/pce.70068","url":null,"abstract":"","PeriodicalId":222,"journal":{"name":"Plant, Cell & Environment","volume":" ","pages":""},"PeriodicalIF":6.0,"publicationDate":"2025-07-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144590116","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"OsPIP2-1 positively regulates rice seed germination by modulating OsAmy3C and α-amylase expression","authors":"Bingxian Chen,&nbsp;Yuanxuan Peng,&nbsp;Qi Zhang,&nbsp;Zhongjian Chen,&nbsp;Hongmei Li,&nbsp;Jun Liu","doi":"10.1111/tpj.70314","DOIUrl":"https://doi.org/10.1111/tpj.70314","url":null,"abstract":"<div>\u0000 \u0000 <p>As timely germination of seeds is crucial to crop yield and quality, the low germination percentage for direct seeding in rice and preharvest sprouting during the late stage of maturity have extremely adverse effects on rice production. Herein, we found that an aquaporin gene <i>OsPIP2-1</i> was significantly expressed during rice seed germination. The <i>OsPIP2-1</i> overexpression seeds showed a higher germination rate, while the knockout lines showed a delayed germination phenotype, indicating that this gene positively regulates seed germination. Furthermore, in seeds of <i>OsPIP2-1</i> overexpression and knockout lines, the activity of α-amylase and the expression of its encoding genes, <i>OsAmy1C</i> and <i>OsAmy1E</i>, were respectively enhanced and suppressed. When <i>OsAmy3C</i> was knocked out in the <i>OsPIP2-1</i> overexpression lines, seed germination was significantly inhibited. Also, the germination of seeds with <i>OsAmy3C</i> knocked out in the wild-type background was delayed, and the expression level of <i>OsPIP2-1</i> in these seeds remained almost unchanged during germination, suggesting that <i>OsPIP2-1</i> is upstream of <i>OsAmy1C</i>. In addition, the ABA-biosynthesis genes <i>OsNCED2/3</i> were reduced in <i>OsPIP2-1</i> overexpression lines but raised in knockouts, while the ABA-catabolism gene <i>OsABA8ox1-3</i> exhibited the opposite trend. Moreover, we have also applied nanosilver, a potential inhibitor of aquaporins, which effectively delays rice seed germination. At the molecular level, the expression of <i>OsPIP2-1</i> in transcriptional and translational levels was significantly decreased by AgNPs. Meanwhile, the expression of α-amylase and its gene expression were also inhibited by AgNPs. In conclusion, <i>OsPIP2-1</i> positively regulates rice seed germination by regulating <i>OsAmy3C</i> and α-amylase. It can enhance seed germination vigor in direct-seeded rice cultivation. Nanosilver delays germination by affecting <i>OsPIP2-1</i> and α-amylase, thus offering a potential strategy for preventing preharvest sprouting.</p>\u0000 </div>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"123 1","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144573659","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Unveiling the effect of curcumin on ion channels of SBMA motoneuron-derived cells and human IPSC-derived neurons: initial electrophysiological findings.","authors":"Vera Plakhova, Ingrid Battistella, Vladimir A Martínez-Rojas, Marta Marchioretto, Daniele Arosio, Linda Masello, Luciano Conti, Carlo Musio","doi":"10.1007/s00249-025-01780-w","DOIUrl":"https://doi.org/10.1007/s00249-025-01780-w","url":null,"abstract":"<p><p>Curcumin (CUR), a bioactive compound extracted from the turmeric (Curcuma longa), has gathered considerable attention in recent years due to its claimed health benefits, including anti-inflammatory, antioxidant, and neuroprotective properties. The dysregulation of ion channel activity and the altered neuronal excitability in neurons has been identified as a key factor in the pathophysiology of neurological disease and a putative pharmacological target for therapeutic options. Therefore, we investigated by whole-cell patch-clamp the CUR's impact on the ionic currents in motoneuron-derived (MN-1) cells modeling SBMA and in human neuro-progenitor-cell (hNPCs)-derived neurons. CUR decreased viability in non-pathological MN-1 cells but showed increased resistance in pathological MN-1 cells, while mature neurons derived from hiPSCs remained unaffected under the same conditions. Electrophysiological studies revealed that CUR inhibits outward and inward currents in both MN-1 cell types, with a more pronounced effect in pathological cells. In hNPC-derived neurons, CUR also inhibited both currents and induced a negative shift in the voltage dependence of activation, suggesting reduced excitability. Our results indicate that further investigations are needed to confirm the role of CUR in the context of neurotherapeutics based on ion channel-targeting pharmacology.</p>","PeriodicalId":548,"journal":{"name":"European Biophysics Journal","volume":" ","pages":""},"PeriodicalIF":2.2,"publicationDate":"2025-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144582775","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"ARF10, ARF16, and ARF17 are involved in cytokinesis during pollen development in Arabidopsis","authors":"Chi Zhang,&nbsp;Keng-Yu Pan,&nbsp;Ben-Shun Zhu,&nbsp;Wen Hu,&nbsp;Yu Jiang,&nbsp;Jie Zhou,&nbsp;Ke Ye,&nbsp;Jia-Jie Chen,&nbsp;Chang-Sheng Mao,&nbsp;Zhong-Nan Yang,&nbsp;Xiaozhen Yao","doi":"10.1111/tpj.70328","DOIUrl":"https://doi.org/10.1111/tpj.70328","url":null,"abstract":"<div>\u0000 \u0000 <p>Pollen formation involves several cell divisions: meiosis I/II and pollen mitosis I/II. Auxin response factors (ARFs) are essential transcription factors in the auxin signaling pathway. While ARFs have been implicated in pollen wall patterning and pollen maturation, their roles in cell division during pollen formation remain unclear. Here, we characterize the essential roles of ARF10, ARF16, and ARF17 in the cytokinesis processes of meiosis and PMI. In the <i>arf10arf16arf17</i>^<i>+/−</i> mutant, the cell plate is abnormally deposited during PMI cytokinesis, resulting in irregular cell identity and pollen degeneration. These phenotypes are consistent with ARF10 and ARF16 being expressed primarily in the vegetative cell of bicellular pollen. The phenotypes of the <i>arf10arf16arf17</i>^<i>+/−</i> mutant are similar to those of <i>glucan synthase-like 10</i> (<i>gsl10</i>), a loss-of-function mutant of a member of the callose synthesis family. <i>GSL10</i> is expressed in the vegetative cell of bicellular pollen, and its transcript is reduced in the <i>arf10arf16arf17</i>^<i>+/−</i> mutant. We confirmed that these three ARFs can directly bind to the <i>GSL10</i> promoter. Thus, we reveal an ARF10, ARF16, and ARF17-GSL10 pathway that is specifically activated in the vegetative cell for pollen development. Moreover, we discovered that in the <i>arf17</i> single mutant, the cell plate was abnormally formed after meiosis I, resulting in cytokinesis defects in the tetrads. Collectively, our results indicate that ARF17 alone is involved in meiotic cytokinesis and that ARF10, ARF16, and ARF17 are redundantly required for PMI cytokinesis. Our research thus provides insights into the key auxin signaling pathways involved in cell division during pollen development.</p>\u0000 </div>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"123 1","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144573660","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"PD-1 expressing islet-specific CD4⁺ T cells promote bystander tolerance and prevent autoimmunity.","authors":"Jeniffer D Loaiza Naranjo, Vivian Zhang, Rathna Ravichandran, Anne-Sophie Bergot, Ranjeny Thomas, Emma E Hamilton-Williams","doi":"10.1111/imcb.70044","DOIUrl":"https://doi.org/10.1111/imcb.70044","url":null,"abstract":"<p><p>Loss of T-cell tolerance to multiple islet antigens is a key feature of autoimmune type 1 diabetes. In this study, we investigated the requirement for programmed death 1 (PD-1) expression by CD4⁺ T cells in the maintenance of self-tolerance via bystander suppression of autoreactive CD8⁺ T cells using nonobese diabetic mice. We used CRISPR/Cas9 to selectively knockout PD-1 in islet antigen-specific BDC2.5 CD4⁺ T cells and observed the impact on bystander tolerance of 8.3 CD8⁺ T cells, specific for a different islet antigen. Loss of PD-1 promoted the proliferation, Th1-like effector-memory phenotype, islet infiltration and expression of cytotoxic markers by BDC2.5 cells. PD-1-deficient BDC2.5 cells were impaired in their regulation of 8.3 cells, which proliferated more, developed an effector-memory phenotype and increased expression of effector molecules. While antigen-presenting cell maturation and migration into the pancreatic lymph node were not impacted by loss of PD-1 expression from BDC2.5 cells, migration of BDC2.5 cells out of the lymph node was required for enhanced activation of the CD8⁺ T cells. Together, these events led to accelerated diabetes progression, suggesting that PD-1 expression by CD4⁺ T cells promotes a tolerogenic microenvironment and restraining autoreactive CD8⁺ T cells.</p>","PeriodicalId":179,"journal":{"name":"Immunology & Cell Biology","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2025-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144582738","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Interplay of brassinosteroids, strigolactones, and CLE44 in modulating Arabidopsis stem architecture in response to mechanical stress","authors":"B. Lorena Raminger,&nbsp;Aitor Muñoz-Gasca,&nbsp;Pilar Cubas,&nbsp;Raquel L. Chan,&nbsp;Julieta V. Cabello","doi":"10.1111/tpj.70329","DOIUrl":"https://doi.org/10.1111/tpj.70329","url":null,"abstract":"<div>\u0000 \u0000 <p>Vascular tissues are crucial to providing plants with physical support and transporting water, nutrients, and signaling metabolites. Mechanical stress produced by wind, insects, and other external factors affects plant growth and development. Mechanical load weight treatments, simulating these stressors, are known to induce specific changes in vascular tissues, leading to increased stem diameter and a higher number of vascular bundles (VBs). In this work, brassinosteroids (BRs) and strigolactones (SLs) are shown as essential for the anatomical changes provoked in the <i>Arabidopsis thaliana</i> stem architecture in response to weight-induced mechanical stress. Unlike wild-type plants, BR signaling mutants (<i>bes1</i> and <i>bzr1</i>) and plants treated with the BR synthesis inhibitor brassinazole failed to exhibit the characteristic increase in stem diameter and VB number after mechanical weight treatment. The SL synthesis gene <i>MAX4</i> and the SL-responsive gene <i>BRC1</i> play a crucial role in stem widening and increasing VB number. Supporting this, <i>max4</i> and <i>brc1</i> mutants neither showed increased stem diameter nor VB number in response to weight treatment. Moreover, <i>CLE44</i>, a downstream target of BRC1, also plays a necessary role, as <i>cle44</i> mutants failed to respond to the weight stimulus. Interestingly, <i>CLE44</i> expression is induced by the synthetic SL analogue GR24 but not by BRs. These findings underscore the convergent and essential roles of BRs and SLs in adapting stem architecture in response to mechanical stress.</p>\u0000 </div>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"123 1","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144573658","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Multi-omics analyses provide insights into the molecular basis for salt tolerance of Phyla nodiflora","authors":"Liyuan Wang,&nbsp;Nan Liu,&nbsp;Yuheng Zhou,&nbsp;Feng Zheng,&nbsp;Shuguang Jian,&nbsp;Xuncheng Liu","doi":"10.1111/tpj.70325","DOIUrl":"https://doi.org/10.1111/tpj.70325","url":null,"abstract":"<div>\u0000 \u0000 <p>The perennial herbaceous plant, <i>Phyla nodiflora</i> (Verbenaceae), which possesses natural resistance to multiple abiotic stresses, is widely used as a pioneer species in island ecological restoration. Due to the lack of information about its genome, the mechanism underlying its tolerance to environmental stresses, such as salinity, is almost entirely unknown. Here, we report on the high-quality genome of <i>P. nodiflora</i> that is 403.07 Mb in size, and which was assembled and anchored onto 18 pseudo-chromosomes. Genomic synteny revealed that <i>P. nodiflora</i> underwent two whole genome duplication events, which promoted the expansion of genes related to environmental adaptation and the biosynthesis of secondary metabolites. An integrated genomic and transcriptomic analysis suggested that salt stress tolerance in <i>P. nodiflora</i> is associated with the expansion and activated expression of genes related to abscisic acid (ABA) homeostasis and signaling. The expansion of ZEP family genes may contribute to the consistent increase in ABA levels under salt stress. Lysine acetylomic analysis revealed that exposure to salt led to widespread protein deacetylation, with these proteins primarily involved in signal transduction, carbohydrate transport and metabolism, and transcription regulation. Deacetylation of glutathione <i>S</i>-transferase increased enzymatic activities in response to salt-induced oxidative stress. Collectively, the genomic, transcriptomic, and lysine acetylomic analyses provide profound insight into the molecular basis of the adaptation of <i>P. nodiflora</i> to salt stress, and will be helpful to engineer salt-tolerant plants for ecological restoration.</p>\u0000 </div>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"123 1","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144573614","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Non-Invasive Phenotyping of Sugar Beet and Maize Roots Using Field-Scale Spectral Electrical Impedance Tomography.","authors":"Valentin Michels, Maximilian Weigand, Lena Lärm, Onno Muller, Andreas Kemna","doi":"10.1111/pce.70049","DOIUrl":"https://doi.org/10.1111/pce.70049","url":null,"abstract":"<p><p>Root systems are essential for plant water and nutrient uptake, but are difficult to characterize in-situ due to their inaccessibility. Spectral electrical impedance tomography (sEIT) is a non-invasive geoelectrical method that has shown potential to quantify root traits at the laboratory scale. However, field applications remain scarce due to technical limitations and challenges in separating soil and root polarization signatures. This study explores the use of sEIT for in-situ phenotyping of sugar beet and maize root systems. We conducted multi-frequency sEIT measurements at varying crop growth stages to derive the subsurface complex resistivity distribution. Spectral analysis revealed high-frequency polarization peaks for both species. Additionally, sugar beets exhibited an additional low-frequency peak late-season, which we attribute to the development of large storage parenchyma. For sugar beet, the high root-to-soil volume fraction allowed a direct correlation of electrical parameters to root biomass density. For maize, the superimposed soil polarization necessitated an alternative approach: We introduce an electrical root index ( <math> <semantics> <mrow><mrow><mi>ERI</mi></mrow> </mrow> <annotation>${ERI}$</annotation></semantics> </math> ) as a spectral dispersion measure indicative of root presence and show its correlation to root biomass density. Our findings demonstrate that sEIT is sensitive to macro- and microscopic root traits under field conditions, holding great potential for non-invasive phenotyping of plant roots.</p>","PeriodicalId":222,"journal":{"name":"Plant, Cell & Environment","volume":" ","pages":""},"PeriodicalIF":6.0,"publicationDate":"2025-07-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144574569","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Biregionally differentiated growth generates sharp apex and concave joints in leaves","authors":"Zining Wang,&nbsp;Yasuhiro Inoue,&nbsp;Atsushi Mochizuki,&nbsp;Hirokazu Tsukaya","doi":"10.1111/tpj.70310","DOIUrl":"https://doi.org/10.1111/tpj.70310","url":null,"abstract":"<p>The leaf apex, the distal end of the leaf blade, exhibits enormous variation in shapes across plant species. Among these diverse morphologies, the sharp apex, characterized by its pointed and elongated tip, is important for both species identification and environmental adaptation. Despite its taxonomic and ecological importance, the developmental mechanisms underlying the formation of a sharp apex remain unknown. The present study aims to investigate the curvature patterns and morphogenesis of the sharp apex to uncover these mechanisms using <i>Triadica sebifera</i> leaves. We revealed that the sharp apex marks the maximum positive curvature and is flanked by concave joints with negative curvatures, indicating anisotropic tissue growth and spatially regulated cellular behavior. To investigate the underlying cellular mechanism, we observed cell shapes and cell divisions across different developmental stages and regions. Unlike plant roots or stems, we did not observe highly elongated or aligned cell shapes at the mature stage. Also, unlike serration leaf margins, we did not observe increased cell proliferation near the sharply elongated apex. Instead, we identified a biregional differentiation in cell division angles, and our simulations confirmed that these division angles could generate the sharply elongated apex that might be influenced by anisotropic cell growth. Then, further generalizations were made from this case study of <i>T. sebifera</i>, revealing that spatiotemporal change in cell division angle is essential to make sharp-tipped leaf shape.</p>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"123 1","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-07-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/tpj.70310","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144563386","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}