Plant Cell Reports最新文献

{"title":"Type-A and -C response regulator genes positively impact rice plant height and panicle architecture.","authors":"Chenyu Rong, Renren Zhang, Jing Xie, Jieru Li, Tiantian Yan, Ziyu Liu, Yuexin Liu, Ruihan Xu, Xi'an Shi, Xuebin Zhao, Jiali Song, Yayi Meng, Zhongyuan Chang, Yanfeng Ding, Chengqiang Ding","doi":"10.1007/s00299-025-03531-9","DOIUrl":"https://doi.org/10.1007/s00299-025-03531-9","url":null,"abstract":"<p><strong>Key message: </strong>Thirteen type -A and two type -C RR genes exhibited distinct expression patterns and unique functions. Notably, RR2 and RR4 had the most significant positive effects impact on panicle development. Cytokinin signal transduction occurs through a \"two-component system\". Type-A and -C response regulators (RRs) are groups of proteins of similar structures constituting significant components of cytokinin signal transduction. In rice, 13 (Type-A) and 2 (Type-C) RRs have been identified to date; however, their functions remain partially known. In this study, we examined the expression patterns of Type-A and Type-C RRs in rice using RNA-Seq and confirmed their functions by constructing mutants of the 15 genes with CRISPR/Cas9. Almost all Type-A RRs played positive roles in the development of secondary branches and secondary spikelets, particularly RR2 and RR4. Notably, rr1 rr2 and rr8 rr12 rr13 higher-order mutants displayed small panicle sizes and decreased plant height. Additionally, both Type-C RRs played positive roles in regulating heading date. RNA-seq revealed several genes with significantly altered expression in the rr2 and rr4 mutants, with almost half of the differentially expressed genes (DEGs) overlapping between the two mutants. Many of the DEGs were associated with the cytokinin and abscisic acid pathways. Our findings provide new insights into the functions of Type-A and -C RRs in rice growth and may serve as a foundation for future studies focusing on cytokinin signaling.</p>","PeriodicalId":20204,"journal":{"name":"Plant Cell Reports","volume":"44 7","pages":"141"},"PeriodicalIF":5.3,"publicationDate":"2025-06-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144310402","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Deciphering ABA/PYL gene family in flax: evolutionary analysis, and abiotic stress response.","authors":"Shashank Kumar Yadav, Ankit Saroha, Devender Singh, Pragya Yadav, Dhammaprakash Pandhari Wankhede, Gyanendra Pratap Singh, Viswanathan Chinnusamy, Vikender Kaur","doi":"10.1007/s00299-025-03517-7","DOIUrl":"https://doi.org/10.1007/s00299-025-03517-7","url":null,"abstract":"<p><strong>Key message: </strong>The identified PYL/PYR/RCAR receptor gene family differentially modulates stress-responsive and developmental roles in linseed. This gene family is evolutionarily and functionally conserved in agronomically imperative oilseed crop species. Abscisic acid (ABA) functions as a key phytohormone, exerting significant regulation over diverse cellular activities, notably growth and responses to various environmental stressors. The protonated form of ABA (ABAH) is perceived by intracellular Pyrabactin resistance 1-like (PYR/PYL) proteins, constituting an integral component of the signaling cascade. The PYLs represent one of the largest phytohormone receptor families identified. The receptors have been extensively investigated in numerous plant species. However, the PYL family in oilseed crop species has not been systematically identified at the genome level. This study identified 15 members of the PYL gene family in the linseed, exhibiting a non-uniform distribution across the 15 chromosomes. Based on structural and functional similarities, the phylogenetic analysis divided linseed PYLs (LuPYLs) into three subfamilies. Gene and motif structure analysis of LuPYLs revealed that members of each subfamily share similar gene and motif structures. An evolutionary relationship with PYL members in other oilseed crop species has been established, wherein whole-genome duplication appears to be the primary driving force underlying the expansion of the PYLs. Collinearity segregated PYL genes from all selected species into five distinct clusters, each representing a conserved functional module. The regulation of PYL genes in linseed, including transcriptional analysis of CREs in promoter regions, identification of SNPs, and assessment of miRNA-mediated post-transcriptional regulation, was assessed. Tissue-specific qRT-PCR analysis revealed diverse expression patterns of LuPYL genes, highlighting their roles in plant development and abiotic stress responses. These findings provide valuable insights into how PYL genes contribute to abiotic stress tolerance in flax.</p>","PeriodicalId":20204,"journal":{"name":"Plant Cell Reports","volume":"44 7","pages":"140"},"PeriodicalIF":5.3,"publicationDate":"2025-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144234928","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Sucrose transport gene FaSWEET9a regulated by FaDOF2 transcription factor promotes sucrose accumulation in strawberry.","authors":"Yan Xu, Shuang Liu, Hongying Sun, Jian Zang, Chao Zhang, Wei Guo, Zhihong Zhang","doi":"10.1007/s00299-025-03528-4","DOIUrl":"10.1007/s00299-025-03528-4","url":null,"abstract":"<p><strong>Key message: </strong>FaSWEET9a, an important sucrose transport gene regulated by transcription factor FaDOF2, regulates the accumulation of sucrose in strawberry fruits and affects the growth of strawberry plants.. This study identified and characterized 25 members of the SWEET gene family in the genome of cultivated strawberry (Fragaria × ananassa cv. 'Yanli'), focusing on their potential roles in fruit development. Notably, FaSWEET9a, a specific member of the SWEET family, was found to be uniquely expressed in 'Yanli' fruit. Functional analysis via heterologous expression in Saccharomyces cerevisiae confirmed that FaSWEET9a acts as a sucrose transporter. To further investigate its role, we generated FaSWEET9a overexpression lines and demonstrated that FaSWEET9a not only enhances sucrose accumulation in strawberry fruits but also influences plant growth and development. We identified FaDOF2 that could bind to the promoter of FaSWEET9a and enhance its transcription by conducting yeast one-hybrid assays, electrophoretic mobility shift assays, β-glucuronidase assays, and luciferase reporter gene assays. Moreover, transient transformation experiments revealed that FaDOF2 could elevate sucrose content in strawberry fruits by regulating FaSWEET9a. This research brings new viewpoints on the molecular mechanisms that govern sucrose regulation in strawberry fruits, spotlighting the functional significance of the FaSWEET9a-FaDOF2 regulatory module in the aspects of fruit quality and development.</p>","PeriodicalId":20204,"journal":{"name":"Plant Cell Reports","volume":"44 6","pages":"138"},"PeriodicalIF":5.3,"publicationDate":"2025-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144199894","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Enhanced thermotolerance via overexpression of a stromal ascorbate peroxidase in Nannochloropsis oceanica.","authors":"Shan Wu, Yiting She, Xitao Chen, Xingwei Huang, Tao Xu, Congcong Miao, Qianyi Wu, Zheng Li, Chengcheng Li, Yuhui Cheng, Yi Xin","doi":"10.1007/s00299-025-03511-z","DOIUrl":"10.1007/s00299-025-03511-z","url":null,"abstract":"<p><strong>Key message: </strong>Overexpression of the heat-responsive ascorbate peroxidase gene NoAPX2430 improves thermotolerance and growth of Nannochloropsis oceanica, offering potential strategies for enhanced resilience in plant cells. Microalgae are promising for industrial lipid production but face challenges from high-temperature (HT) stress. This study focused on improving thermotolerance in Nannochloropsis oceanica by studying and modifying the heat-responsive ascorbate peroxidase gene, NoAPX2430. Functional and localization analyses confirmed NoAPX2430 as an active enzyme in the chloroplast stroma. Knockdown of NoAPX2430 resulted in reduced growth, photosynthetic efficiency, and reactive oxygen species (ROS) scavenging under HT (35 °C). In contrast, overexpression of NoAPX2430 significantly improved photosynthetic performance, lowered ROS levels, and boosted growth rates. Additionally, lipid and triacylglycerol contents increased significantly under combined nitrogen depletion and HT, with polyunsaturated fatty acids rising by up to 194.4%. These results highlight NoAPX2430's critical role in enhancing thermotolerance and lipid biosynthesis. This provides a foundation for genetic and environmental strategies to boost microalgal resilience and productivity for sustainable biofuel and nutraceutical development.</p>","PeriodicalId":20204,"journal":{"name":"Plant Cell Reports","volume":"44 6","pages":"139"},"PeriodicalIF":5.3,"publicationDate":"2025-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144199893","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Dual regulation of seed coat structure and flavonoids biosynthesis on seed dormancy formation of alfalfa.","authors":"Shushuan Wang, Yutong Zhang, Ruru Shi, Wenqiang Fan, Fengling Shi","doi":"10.1007/s00299-025-03526-6","DOIUrl":"10.1007/s00299-025-03526-6","url":null,"abstract":"<p><strong>Key message: </strong>The metabolic regulation of flavonoids and GAs reinforces the defensive function of the seed coat, thereby establishing and maintaining dormancy. Medicago falcata L. is an important genetic resource for stress resistance in legumes with high seed value. However, its hard seed coat causes low germination rates, uneven seedling emergence, and prolonged dormancy. Moreover, the contributions of seed coat structure and secondary metabolites, including flavonoids and phytohormones, to dormancy formation in Medicago falcata L. seeds remain unknown. This study investigated structural changes in the seed coat surface and internal layers, along with accumulation patterns of flavonoids and phytohormone biosynthesis/catabolism. Results showed that hard seeds had extremely low germination rates, rough seed coat surfaces, reduced hilum aperture size, and densely arranged internal palisade layers with maximum thickness, confirming the decisive role of seed coat structure (especially the palisade layer) in dormancy. Flavonoids, such as flavonols and isoflavonoids (liquiritigenin, myricetin, daidzein, 3-O-methylquercetin, and kaempferol-3-O-galactoside), were significantly upregulated in hard seeds. Notably, anthocyanin and flavonol biosynthesis share the common precursor aromadendrin, which was preferentially allocated toward flavonol production, leading to the downregulation of anthocyanin precursors (dihydroquercetin, afzelechin, epiafzelechin). Gibberellins (GA4, GA29, GA34) were significantly downregulated in hard seeds, while elevated ABA/GAs and ABA/JA ratios promoted dormancy formation. This study reveals changes in seed coat texture, internal cell layer arrangement, and palisade layer thickness during hard seed development, while highlighting the core regulatory roles of flavonoids and GAs in dormancy establishment.</p>","PeriodicalId":20204,"journal":{"name":"Plant Cell Reports","volume":"44 6","pages":"137"},"PeriodicalIF":5.3,"publicationDate":"2025-05-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144187804","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Insights into the roles of chromatin-remodeling complex HOS15-PWR-HDA9 in plant development and stress responses.","authors":"Akhtar Ali, Shah Zareen, Dae-Jin Yun","doi":"10.1007/s00299-025-03523-9","DOIUrl":"10.1007/s00299-025-03523-9","url":null,"abstract":"<p><strong>Key message: </strong>This review highlights the central role of the HOS15-PWR-HDA9 complex in integrating chromatin remodeling with plant development and stress adaptation, offering insights for improving crop resilience. Being sessile by nature, plants have signaling pathways that perceive and transmit environmental signals, inducing epigenetic modifications that are instrumental in plant development and stress responses. Acetylation and methylation are key features of chromatin remodeling and are crucial in transcriptional reprogramming for tissue-specific gene expression and silencing. Recently, different classes of chromatin remodelers have been studied in plants. Among them, the multiprotein complex HOS15-PWR-HDA9 has been well documented to regulate the acetylation status of histone 3 at the chromatin of target genes during plant development and in response to various stresses, thereby facilitating gene expression and/or repression. In this review, we discuss the critical role of the HOS15-PWR-HDA9 complex in chromatin remodeling during plant development and stress responses.</p>","PeriodicalId":20204,"journal":{"name":"Plant Cell Reports","volume":"44 6","pages":"134"},"PeriodicalIF":5.3,"publicationDate":"2025-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144174700","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"VvWUS increases carpel number by upregulating VvAG2 expression in grapevine.","authors":"Jiang Wu, Zhenhua Liu, Jianfang Hu, Pingyin Guan","doi":"10.1007/s00299-025-03522-w","DOIUrl":"10.1007/s00299-025-03522-w","url":null,"abstract":"<p><strong>Key message: </strong>The positive effect of VvWUS on carpel number variation in Vitis vinifera 'Xiangfei' is accomplished via forming the heterodimer with VvSTMa and VvSTMb proteins, which enhances the downstream VvAG2 gene expression during grapevine carpel formation. The WUSCHEL-CLAVATA3 signalling pathway plays an essential role in the shoot apical meristem (SAM) maintenance and floral organ development, influencing carpel number and fruit morphology in horticultural plants. This study investigates the role of VvWUS in regulating carpel number in the grapevine cultivar 'Xiangfei', which frequently produces flowers with multiple carpels. We found a positive correlation between VvWUS expression levels and carpel number variation among grapevine cultivars. Transgenic tomato plants overexpressing VvWUS exhibited an increased carpel number and a concomitant expression of SAM-related genes. Further experiments demonstrated that VvWUS interacts with VvSTMa and VvSTMb, forming heterodimers that are crucial for grapevine carpel development. VvWUS was also confirmed to activate VvAG2 expression. Our findings indicate that VvWUS positively regulates carpel number in grapevine by interacting with VvSTMa and VvSTMb and upregulating VvAG2 expression. These results provide a basis for molecular breeding aimed at increasing grape yield and fruit size.</p>","PeriodicalId":20204,"journal":{"name":"Plant Cell Reports","volume":"44 6","pages":"136"},"PeriodicalIF":5.3,"publicationDate":"2025-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144182432","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Exogenous auxin-induced rapid modulation of herbaceous peony stem straightness: insights into transient cell wall metabolism responses.","authors":"Yiran Huang, Yijia Jin, Mengting Zhu, Wanning Guo, Tianyu Zhao, Siying Chen, Yingling Wan, Yan Liu","doi":"10.1007/s00299-025-03525-7","DOIUrl":"10.1007/s00299-025-03525-7","url":null,"abstract":"<p><strong>Key message: </strong>Exogenous IAA applying within 32 days after herbaceous peony bud germination induces a 6-h transient stem change: first bending then upright. This process correlates with the degradation of cell wall substances like cellulose and pectin. Herbaceous peony (Paeonia lactiflora) holds a growing presence in the cut flower market. A vital criterion for selecting cut herbaceous peonies is the uprightness of their stems, which is largely determined by cell wall materials (CWMs). Our previous study indicated that applying indole-3-acetic acid (IAA) during the development process could enhance the stem straightness of herbaceous peonies at the flowering stage. Interestingly, in certain development phases, after IAA application, the stems were observed to first bend and then regain an upright position within hours (transient changes), yet the underlying mechanism remains unclear. This study aimed to identify the stages during which these transient changes occur and elucidate the role of CWMs in terms of their contents, enzyme activities, metabolites, and gene expression. The results showed that IAA-induced transient changes were most prevalent in the first three stages. The accumulated cellulose, pectin, and lignin in the IAA-treated group were consumed as the stem regained their upright position. Cinnamyl-alcohol dehydrogenase, β-glucosidase, and pectin methylesterase played key degradation roles. Combined metabolome and transcriptome analyses revealed that differential metabolites and differentially expressed genes were enriched in pathways such as glycolysis/gluconeogenesis, xylan biosynthesis process, secondary cell wall biogenesis, and lignin catabolism. The cellulose synthesis gene CESA2, decomposition gene CEL5, and pectin decomposition gene At1g48100 deserved further investigation. This study provides support for clarifying the mechanism by which IAA regulates stem uprightness of P. lactiflora and serves as a reference for selection and cultivation of herbaceous peony cut flowers.</p>","PeriodicalId":20204,"journal":{"name":"Plant Cell Reports","volume":"44 6","pages":"135"},"PeriodicalIF":5.3,"publicationDate":"2025-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144174696","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A novel-miR-164 from Atractylodes lancea targets AlNAC1 regulating leaf senescence.","authors":"Juan Deng, Xiao Huang, Meng Wang, Qian Li, Ling Gong, Bisheng Huang, Kun Yu","doi":"10.1007/s00299-025-03527-5","DOIUrl":"10.1007/s00299-025-03527-5","url":null,"abstract":"<p><strong>Key message: </strong>A novel miR164-AlNAC1 module adjusting the expression of genes related to chlorophyll synthesis and ROS synthesis, and regulating leaf senescence in Atractylodes lancea. Leaf senescence is a critical developmental process regulated by complex genetic networks. In this study, a novel miRNA belonging to the MIR164 family was identified as a key regulator of leaf senescence in A. lancea by targeting the transcription factor AlNAC1. Bioinformatics analysis revealed that target gene AlNAC1 was closely related to the senescence-associated gene ANAC092 in Arabidopsis. The qRT-PCR and tobacco transient expression demonstrated nov-miR164 negatively regulated AlNAC1. Subcellular localization and transcriptional activity assays revealed that AlNAC1 localized to the nucleus and functioned as a transcriptional activator. Furthermore, EMSA experiments showed that AlNAC1 bound to the G-box motifs on the promoters of AlNYC, AlPAO, and AlRbohC. With the aging of A. lancea leaves, the expression of nov-miR164 decreased, while AlNAC1 and senescence-associated genes (AlSAG21, AlNYC, AlPAO, and AlRbohC) were significantly upregulated. Agrobacterium-mediated transient expression experiment results demonstrated that nov-miR164 negatively regulated AlNAC1 to suppress chlorophyll degradation and ROS accumulation, thereby delaying leaf senescence. Our findings provide new insights into the molecular mechanisms of leaf senescence in A. lancea and highlight the potential of nov-miR164 as a target for increasing crop yield and improving crop longevity.</p>","PeriodicalId":20204,"journal":{"name":"Plant Cell Reports","volume":"44 6","pages":"133"},"PeriodicalIF":5.3,"publicationDate":"2025-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144174695","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Ovule development and pollen tube growth in Tsuga chinensis: insights into the evolution of siphonogamy.","authors":"Xin Shang, Ya-Xin Wang, Guo-Wei Jiang, Zhi-Han Chen, Xiao-Hang Bi, Jie Sun, Xin Zhang","doi":"10.1007/s00299-025-03519-5","DOIUrl":"10.1007/s00299-025-03519-5","url":null,"abstract":"<p><strong>Key message: </strong>Confirmation of the correlation between archegonium development and pollen tube guidance and identification of putative genes implicated in male‒female interactions in gymnosperms. Studying angiosperm-like siphonogamy in gymnosperms will increase our understanding of seed plant evolution. This study involved an exploration of pollination and ovule development in Tsuga chinensis, which is an interesting gymnosperm species endemic to East Asia that exhibits angiosperm-like siphonogamy. Using comprehensive morphological approaches, we determined that the mechanism underlying T. chinensis pollination involves the germination of pollen grains outside the ovule and the growth of pollen tubes into the micropyle. Furthermore, a correlation between the timing of archegonial appearance and pollen tube growth was confirmed. Additionally, transcriptome sequencing revealed 19,643 DEGs that are involved in ovule development. Through bioinformatics analysis, we identified several putative genes that are involved in male‒female interactions in gymnosperms, and further validation of the functions of these DEGs is worthwhile. These findings offer valuable insights into the progression of the complex evolution of siphonogamy across seed plants and lay a foundation for understanding the molecular mechanisms underlying siphonogamy in gymnosperms.</p>","PeriodicalId":20204,"journal":{"name":"Plant Cell Reports","volume":"44 6","pages":"132"},"PeriodicalIF":5.3,"publicationDate":"2025-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144143352","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}