Valentina Vallejos, Francisca Fuentes, Domingo Sancho-Knapik, Jorge Gago, Constanza F Ramírez, Betsy K Rivera, Lohengrin A Cavieres, Jeroni Galmés, José Javier Peguero-Pina, Eustaquio Gil-Pelegrín, Patricia L Sáez
{"title":"Hydraulic and Photosynthetic Performance of Antarctic Plants Under Successive Freeze-Thaw Cycles.","authors":"Valentina Vallejos, Francisca Fuentes, Domingo Sancho-Knapik, Jorge Gago, Constanza F Ramírez, Betsy K Rivera, Lohengrin A Cavieres, Jeroni Galmés, José Javier Peguero-Pina, Eustaquio Gil-Pelegrín, Patricia L Sáez","doi":"10.1111/pce.15528","DOIUrl":"https://doi.org/10.1111/pce.15528","url":null,"abstract":"<p><p>Climate change projections predict warming and increased weather variability, mainly in polar regions, altering freeze-thaw patterns. However, the effects of rising temperatures and more frequent freeze-thaw events on the water and CO<sub>2</sub> management of Antarctic plants remain unclear. To address this, we conducted a laboratory experiment to investigate how growth temperature (5°C and 15°C) and successive freeze-thaw cycles influence the hydraulic and photosynthetic performance of Deschampsia antarctica (D. antarctica) and Colobanthus quitensis (C. quitensis). Our results showed that warmer conditions improved hydraulic and photosynthetic performance in both species, driven by anatomical adjustments in leaf xylem vessels. Additionally, plants exposed to successive freeze-thaw cycles exhibited a coordinated decline in whole-plant hydraulic conductivity and leaf gas exchange, regardless of growth temperature. The magnitude of changes (%) in photosynthetic traits after freeze-thaw cycles varied between species, with D. antarctica showing similar responses at both growth temperatures, while C. quitensis experienced more pronounced changes at the lower temperature. Overall, these findings suggest that while Antarctic plants benefit from warmer temperatures, repeated freeze-thaw events could disrupt their hydraulic balance and limit photosynthesis, particularly under natural environmental conditions.</p>","PeriodicalId":222,"journal":{"name":"Plant, Cell & Environment","volume":" ","pages":""},"PeriodicalIF":6.0,"publicationDate":"2025-04-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143802068","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}
Fengwei Gu, Huabin Xie, Qiwei Huang, Wenjie Zhou, Xiaodi Zou, Zhikai Han, Tao Guo, Hui Wang, Jiafeng Wang
{"title":"Co-Expression Pattern Analysis of Head-to-Head NLR Gene Pair Pik-H4.","authors":"Fengwei Gu, Huabin Xie, Qiwei Huang, Wenjie Zhou, Xiaodi Zou, Zhikai Han, Tao Guo, Hui Wang, Jiafeng Wang","doi":"10.1111/pce.15509","DOIUrl":"https://doi.org/10.1111/pce.15509","url":null,"abstract":"<p><p>Nucleotide-binding leucine-rich repeat (NLR) genes play a critical role in plant effector-triggered immunity (ETI) against pathogen invasion. However, the regulatory mechanisms governing NLR expression and functional dynamics, particularly in head-to-head NLR gene pairs, remain poorly understood. In this study, we investigated the regulatory mechanisms, subcellular localization and functional pathways associated with Pik-H4 gene pair. Bidirectional Pik-H4 promoter (P<sub>Pik-H4</sub>) strengths were found across the whole plants and exhibited co-expressed patterns in tissues and cells, and the P<sub>Pik-H4</sub> activity was upregulated in vascular bundles during blast fungus invasion. Additionally, altering the co-expression of Pik<sub>1</sub>-H4 and Pik<sub>2</sub>-H4 via overexpression in rice or Nicotiana benthamiana did not compromise the immune response. Promoter analysis identified two minimal promoter regions that are essential for bidirectional transcription, and mutagenesis of the bidirectional TATA box confirmed its role in gene regulation. This dual-function promoter coordinates Pik-H4 expression in both directions, a regulatory innovation previously unreported in NLR-mediated immunity. In planta subcellular localization revealed Pik<sub>1</sub>-H4 relocates to vesicles, indicating its role in effector recognition, while Pik<sub>2</sub>-H4 predominantly accumulated in the nucleus. These new discoveries of Pik protein extended the putative immune function of NLR pairs. Transcriptome analysis demonstrated that Pik-H4-mediated resistance induces significant transcriptome reprogramming between 12- and 24-h postinoculation. In summary, these findings provide novel insights into the regulatory complexity and functional divergence within NLR bidirectional gene pairs in response to pathogen invasion.</p>","PeriodicalId":222,"journal":{"name":"Plant, Cell & Environment","volume":" ","pages":""},"PeriodicalIF":6.0,"publicationDate":"2025-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143770845","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}
Margaux Didion-Gency, Alice Gauthey, Kate M Johnson, Philipp Schuler, Charlotte Grossiord
{"title":"Leaf Excision and Exposure Duration Alter the Estimates of the Irreversible Photosynthetic Thermal Thresholds.","authors":"Margaux Didion-Gency, Alice Gauthey, Kate M Johnson, Philipp Schuler, Charlotte Grossiord","doi":"10.1111/pce.15521","DOIUrl":"https://doi.org/10.1111/pce.15521","url":null,"abstract":"<p><p>Understanding plant heat tolerance requires assessing their thermal thresholds, but commonly used methods have rarely been compared. Moreover, whether the photosynthetic machinery is irreversibly damaged past these thresholds remains unclear. We determined the critical temperature (T<sub>crit</sub>), the temperature causing a 50% reduction (T<sub>50</sub>), and the maximum tolerable temperature (T<sub>max</sub>) of photosystem II in Mediterranean cypress, Aleppo pine, and Scots pine saplings using 15- or 30-min heat exposure curves performed on living plants (in-vivo), excised needles (ex-vivo), and excised needles continuously exposed to each rising temperature (ex-vivo continuous). Dark-adapted fluorescence (F<sub>v</sub>/F<sub>m</sub>) and gas exchange were recorded for 4 days postheat stress to track recovery. Longer heat exposure (30 vs. 15 min) consistently led to lower F<sub>v</sub>/F<sub>m</sub>, T<sub>50</sub>, and T<sub>max</sub>. T<sub>50</sub> and T<sub>max</sub> were reduced in both ex-vivo conditions compared to in-vivo ones. Conversely, T<sub>crit</sub> remained consistent between species, exposure durations, and methods. Gas exchange and F<sub>v</sub>/F<sub>m</sub> recovery mainly occurred before reaching T<sub>50</sub> values (about 45°C). Our work highlights the importance of exposure duration and method selection when measuring and comparing thermal thresholds. Moreover, while T<sub>crit</sub> appears to be a reversible threshold, the photosynthetic machinery of studied species appears irreparably damaged past their T50.</p>","PeriodicalId":222,"journal":{"name":"Plant, Cell & Environment","volume":" ","pages":""},"PeriodicalIF":6.0,"publicationDate":"2025-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143770851","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":"Sri Lankan cassava mosaic virus Silencing Suppressor AC4 Mediates Autophagic Degradation of SGS3/RDR6 Bodies in Plants.","authors":"Xueting Liu, Hua Kong, Linyu Liu, Qiuxian Xie, Yan Fu, Xiaoling Yu, Wenbin Li, Yanli Ren, Mengbin Ruan, Xiuchun Zhang","doi":"10.1111/pce.15511","DOIUrl":"https://doi.org/10.1111/pce.15511","url":null,"abstract":"<p><p>RNA silencing and autophagy play crucial roles in maintaining cellular homeostasis and defending against viral infections in diverse eukaryotic hosts. In response to RNA silencing defenses, the majority of plant viruses have evolved viral suppressors of RNA silencing (VSRs). Cassava mosaic geminiviruses (CMGs) are a group of bipartite begomoviruses that cause significant losses to the staple food crop cassava (Manihot esculenta Crantz). The AC4 protein, encoded by CMGs, is a well-characterized VSR; however, the precise mechanism underlying its suppression of RNA silencing remains unclear. This study demonstrates that AC4 expression impedes tasiRNA biogenesis. Moreover, the AC4 protein has been observed to interact with both SGS3 and its partner RDR6, which are essential for the synthesis of trans-acting small interfering RNAs and the amplification of RNA silencing. Notably, these interactions do not disrupt the association between AtSGS3 and AtRDR6 but instead induce their degradation. Furthermore, the AC4-mediated degradation of AtSGS3 is suppressed by an autophagy inhibitor, and AC4 enhances autophagy activity. The results indicate that the autophagy pathway is involved in AC4-mediated degradation of SGS3. These findings reveal a previously unidentified mechanism by which AC4 exploits autophagy to attenuate host RNA silencing, thereby impacting plant development and fulfilling its VSR function. This study offers new insights into the intricate relationship between RNA silencing and autophagy.</p>","PeriodicalId":222,"journal":{"name":"Plant, Cell & Environment","volume":" ","pages":""},"PeriodicalIF":6.0,"publicationDate":"2025-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143778705","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":"DNA Methylation in CYP82E4 Regulates Nicotine Conversion of Nicotiana tabacum.","authors":"Yaqi Wang, Xingzi Zhang, Fang Zhang, Lirui Cheng, Caihong Jiang, Aiguo Yang, Fengxia Li","doi":"10.1111/pce.15520","DOIUrl":"https://doi.org/10.1111/pce.15520","url":null,"abstract":"<p><p>Nornicotine content is very low in tobacco, accounting for less than 5% of total alkaloids. Nicotine conversion refers to the process by which nornicotine is synthesised spontaneously and in large quantities from nicotine. CYP82E4 is the only key enzyme gene involved in nicotine conversion, but it is unclear by what mechanism plants regulate the expression of this gene and thus change the phenotype of nicotine conversion. By comparing single-base resolution DNA methylomes of senescent leaves from NC-L and its high converter variant NC-H, we found two differentially methylated regions (DMRs) in CYP82E4 of NC-H. The bisulfite sequencing PCR (BSP) assay demonstrated that the DNA methylation levels in two specific segments of CYP82E4 were 39%-52% lower for NC-H than for NC-L. Furthermore, treatment with the DNA methylase inhibitor 5-azacitidine resulted in a decrease in DNA methylation levels of CYP82E4 and the change of nicotine conversion phenotype from norconverter tobacco to high converter tobacco. Similarly, the MET1 mutation significantly reduced the DNA methylation level of CYP82E4 and transformed the nicotine conversion phenotype. These findings suggest that DNA methylation plays a crucial regulatory role in nicotine conversion, with decreased methylation levels in CYP82E4 being significant factors in nicotine conversion.</p>","PeriodicalId":222,"journal":{"name":"Plant, Cell & Environment","volume":" ","pages":""},"PeriodicalIF":6.0,"publicationDate":"2025-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143778704","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":"Maternal Temperature Imposes a Longer-Term Effect on Seedling Emergence Than Does Genetic Variation in Seed Dormancy.","authors":"Toshiyuki Imaizumi, Kentaro Ohigashi, Akira Koarai","doi":"10.1111/pce.15525","DOIUrl":"https://doi.org/10.1111/pce.15525","url":null,"abstract":"<p><p>Germination represents the first major transition in plants, and seed dormancy influences germination timing. However, the mechanism by which variations in seed dormancy due to genetic variation or the maternal environment influence germination timing has not been studied in depth. In this study, the effects of temperature during seed maturation (maternal temperature) and genetic variation on weedy rice seedling emergence in a field environment were evaluated. The experiments were repeated for 4 years using seeds collected from weedy rice groups, which represented different degrees of seed dormancy. The maternal temperature was evaluated via the yearly variation in the field temperature. Genetic variation had a greater effect on seedling emergence during unfavourable seasons than during favourable seasons. A higher maternal temperature delayed seedling emergence during favourable seasons. The notable impact of global warming on seedling emergence has been confirmed over the past 15 years, and this impact will continue even under the sustainable CO<sub>2</sub> emission scenario. Maternal effects have long-term effects on seedling emergence at relatively high maternal temperatures, and these effects may increase under global warming.</p>","PeriodicalId":222,"journal":{"name":"Plant, Cell & Environment","volume":" ","pages":""},"PeriodicalIF":6.0,"publicationDate":"2025-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143762565","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":"Outside Front Cover Image","authors":"Sanyan Lai, Wenzhuo Wang, Tianlin Shen, Xiu Li, Dexu Kong, Xiaohan Hou, Gao Chen, Liping Gao, Tao Xia, Xiaolan Jiang","doi":"10.1111/pce.15541","DOIUrl":"https://doi.org/10.1111/pce.15541","url":null,"abstract":"<p>Outside Front Cover: The cover image is based on the article <i>Crucial Role of Aluminium-Regulated Flavonol Glycosides (F2-Type) Biosynthesis in Lateral Root Formation of Camellia sinensis</i> by Sanyan Lai et al., https://doi.org/10.1111/pce.15372.\u0000\u0000 <figure>\u0000 <div><picture>\u0000 <source></source></picture><p></p>\u0000 </div>\u0000 </figure></p>","PeriodicalId":222,"journal":{"name":"Plant, Cell & Environment","volume":"48 5","pages":"i"},"PeriodicalIF":6.0,"publicationDate":"2025-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/pce.15541","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143749722","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}
{"title":"Intercropping Reduces Root Pest Damage via Repellent Volatile Compounds: Insights From Behavioural Assays and Transcriptomic Analysis.","authors":"Xianqin Wei, Chaoying Chen, Xiaorui He, Yuchen Li, Penghua Bai, Ting Liu, Weibin Ruan, Sergio Rasmann","doi":"10.1111/pce.15510","DOIUrl":"https://doi.org/10.1111/pce.15510","url":null,"abstract":"<p><p>Terrestrial plants naturally produce chemical signals to attract beneficial insects or repel harmful pests. These inherent plant attributes offer promising opportunities for eco-friendly pest control in agriculture, particularly through the push-pull intercropping technique. However, our understanding of potential repellent plants and their effective chemical signals remains limited. In this study, we evaluated multiple plant species for their repellent properties, identified effective volatile organic compounds, and investigated the mechanisms for controlling the fungus gnat Bradysia odoriphaga in Chinese chives. Among the 12 species tested, Mentha haplocalyx, Ocimum basilicum and Pelargonium graveolens demonstrated strong repellent effects, making them promising candidates as 'push' plants. Eight major volatile compounds were identified as effective repellents, with 1,8-cineole being the most efficient. 1,8-cineole consistently exhibited repellent effects against the fungus gnats across various concentrations and exposure durations. Transcriptomic analysis revealed that exposure to 1,8-cineole upregulated genes is associated with energy production processes, suggesting that the fungus gnats can detect and actively avoid this compound. Field experiments further confirmed the effectiveness of this strategy, as intercropping chives with M. haplocalyx significantly reduced fungus gnat infestations. This study presents a novel intercropping approach for managing fungus gnats and offers valuable insights into sustainable eco-friendly pest management practices in agriculture.</p>","PeriodicalId":222,"journal":{"name":"Plant, Cell & Environment","volume":" ","pages":""},"PeriodicalIF":6.0,"publicationDate":"2025-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143762563","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}
Lei Wang, Yi-Ping Shi, Yan-Ni Tang, Bao-Shan Xian, Xiao-Tong Ren, Meng-Yuan Ren, Juan He, Yong-Chang Liu, Quan-Le Xu, Peng Chen, Kai Shu
{"title":"The Tartary Buckwheat FtMYB46-FtNRAMP3 Module Enhances Plant Lead and Cadmium Tolerance.","authors":"Lei Wang, Yi-Ping Shi, Yan-Ni Tang, Bao-Shan Xian, Xiao-Tong Ren, Meng-Yuan Ren, Juan He, Yong-Chang Liu, Quan-Le Xu, Peng Chen, Kai Shu","doi":"10.1111/pce.15518","DOIUrl":"https://doi.org/10.1111/pce.15518","url":null,"abstract":"<p><p>The presence of toxic heavy metals lead (Pb) and cadmium (Cd) in polluted soil damage crop production and consequently harms human and livestock health. Tartary buckwheat (Fagopyrum tataricum) is a potential model plant for heavy metal phytoremediation because of its valuable characteristics of high heavy metal tolerance and abundant biomass production. Here, we report that the Tartary buckwheat FtMYB46-FtNRAMP3 module enhances plant Pb and Cd tolerance. RNA sequencing analysis showed that Pb treatment specifically induced expression of FtNRAMP3, a member of the NRAMP (Natural Resistance-Associated Macrophage Protein) transporter gene family. Further cytological and biochemical analysis revealed that FtNRAMP3 was localised to the plasma membrane and significantly contributed to increased tolerance to Pb and Cd in yeast cells. Consistently, transgenic overexpression of FtNRAMP3 in Arabidopsis significantly increased plant tolerance to Pb and Cd applications, reducing Pb concentration but increasing Cd concentration in the overexpression transgenic plants. Subsequent yeast one-hybrid and electrophoretic mobility shift assays showed that the transcription factor FtMYB46 directly binds to the FtNRAMP3 promoter. Further, FtMYB46 promoted FtNRAMP3 expression and increased plant Pb and Cd tolerance. Overall, this study demonstrates the important role of the FtMYB46-FtNRAMP3 module and its potential value in the phytoremediation of Pb and Cd stress.</p>","PeriodicalId":222,"journal":{"name":"Plant, Cell & Environment","volume":" ","pages":""},"PeriodicalIF":6.0,"publicationDate":"2025-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143762572","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}
Shoudong Zhang, Zhixia Xiao, Ailin Liu, Dongpeng Ren, Shengjie Chen, Hanxue Zhang, Li Zhang, Zhili Wang, Jun Yang, Hon-Ming Lam
{"title":"Salt Stress Adaptations in Soybean Involve Alterations in Pre-mRNA Processing.","authors":"Shoudong Zhang, Zhixia Xiao, Ailin Liu, Dongpeng Ren, Shengjie Chen, Hanxue Zhang, Li Zhang, Zhili Wang, Jun Yang, Hon-Ming Lam","doi":"10.1111/pce.15515","DOIUrl":"https://doi.org/10.1111/pce.15515","url":null,"abstract":"<p><p>Salt stress can seriously affect plant survival. To adapt to salt stress, plants can alter gene expressions and/or pre-mRNA processing patterns, or both. Previous studies could not comprehensively profile stress-responsive pre-mRNA processing patterns due to limitations in traditional sequencing technologies. Now Oxford Nanopore Technologies Direct RNA Sequencing (ONT DRS) can directly sequence full-length native RNAs without requiring reverse transcription or amplification. Thus, it provides accurate profiles of pre-mRNA processing patterns at the single-molecule level. With this technology, we found more than 89 586 novel transcript isoforms in addition to the 44 877 annotated ones in soybean leaves and roots subjected to short-term salt stress. Specifically, we identified 102 191 alternative mRNA processing events and 1216 fusion transcripts corresponding to 549 genomic regions. Interestingly, genes upregulated in roots due to salt stress had longer poly(A) tail lengths and lower m6A modification ratios than controls, and downregulated genes in roots had shorter poly(A) tails. Also, the m6A modification levels changed with prolonged salt stress. Furthermore, the alteration patterns of m6A modifications under salt stress were correlated with the expressions of two m6A erasers. Our results indicated that the reshaped mRNA traits caused by salt stress could play a role in soybean adaptations.</p>","PeriodicalId":222,"journal":{"name":"Plant, Cell & Environment","volume":" ","pages":""},"PeriodicalIF":6.0,"publicationDate":"2025-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143770832","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}