{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T12:19:57Z","timestamp":1787660397411,"version":"build-2736575974"},"reference-count":50,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2026,8,12]],"date-time":"2026-08-12T00:00:00Z","timestamp":1786492800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32360255"],"award-info":[{"award-number":["32360255"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32471746"],"award-info":[{"award-number":["32471746"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"award":["32360255"],"award-info":[{"award-number":["32360255"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]},{"award":["32471746"],"award-info":[{"award-number":["32471746"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Plants and their endophytic microbiota form a functional holobiont, wherein host-dependent endophytes (HDEs) are proposed to serve as persistent microbial components associated with host continuity. Using long-term in vitro grapevine plantlets, we investigated HDE redistribution and reassembly across successive agar-based subcultures and during the transition to soil-based cultivation. Results showed that the initial tissue niche (shoot-tip versus root-base) imprinted a persistent compositional signature on HDE communities, with distinct cultivar-dependent effects. Numerous microbial amplicon sequence variants (ASVs) remained shared across agar-based and soil-based habitats, demonstrating high persistence of HDE-associated lineages. Bacterial communities exhibited marked plasticity, with plant-associated bacterial lineages contributing substantially to soil community assembly. Notably, 204 root-enriched bacterial ASVs, comprising 10.8\u201314.3% of the soil community, were recovered, revealing a robust \u201cRoot-Specific Legacy\u201d. Bacterial HDEs showed higher redistribution potential (43.3%), whereas fungal transfer was significantly lower (10.4%). Functional profiling suggested habitat-associated functional patterns, with the anaerobic phenotype enriched in soil and predicted enhancements in membrane transport, nucleotide metabolism, glycan biosynthesis, and biosynthesis of secondary metabolites in roots. Cladosporium and Fusarium remained dominant in the fungal core taxa, and habitat transition altered their predicted trophic profiles. Our findings establish HDEs as a resilient biological reservoir capable of niche-specific expansion, contributing to the ecological reassembly of the plant holobiont.<\/jats:p>","DOI":"10.3390\/microorganisms14081780","type":"journal-article","created":{"date-parts":[[2026,8,12]],"date-time":"2026-08-12T12:53:48Z","timestamp":1786539228000},"page":"1780","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Host-Dependent Endophytes as a Resilient Biological Reservoir: Niche-Specific Expansion and Holobiont Reassembly in Grapevine"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-9779-0065","authenticated-orcid":false,"given":"Jing-Xiu","family":"Tang","sequence":"first","affiliation":[{"name":"Yunnan Key Laboratory for Plateau Mountain Ecology and Restoration of Degraded Environments & School of Ecology and Environmental Sciences, Yunnan University, Kunming 650504, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu-Tao","family":"Wang","sequence":"additional","affiliation":[{"name":"Yunnan Key Laboratory for Plateau Mountain Ecology and Restoration of Degraded Environments & School of Ecology and Environmental Sciences, Yunnan University, Kunming 650504, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hong-Yan","family":"Hu","sequence":"additional","affiliation":[{"name":"Yunnan Key Laboratory for Plateau Mountain Ecology and Restoration of Degraded Environments & School of Ecology and Environmental Sciences, Yunnan University, Kunming 650504, China"},{"name":"Institute of International Rivers and Ecological Security, Yunnan University, Kunming 650504, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jia-Xin","family":"Zhou","sequence":"additional","affiliation":[{"name":"Yunnan Key Laboratory for Plateau Mountain Ecology and Restoration of Degraded Environments & School of Ecology and Environmental Sciences, Yunnan University, Kunming 650504, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rui-Yu","family":"Yang","sequence":"additional","affiliation":[{"name":"Yunnan Key Laboratory for Plateau Mountain Ecology and Restoration of Degraded Environments & School of Ecology and Environmental Sciences, Yunnan University, Kunming 650504, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-1866-7931","authenticated-orcid":false,"given":"Qiu-Yue","family":"Zhang","sequence":"additional","affiliation":[{"name":"Yunnan Key Laboratory for Plateau Mountain Ecology and Restoration of Degraded Environments & School of Ecology and Environmental Sciences, Yunnan University, Kunming 650504, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-8022-3379","authenticated-orcid":false,"given":"Hao","family":"Sun","sequence":"additional","affiliation":[{"name":"Yunnan Key Laboratory for Plateau Mountain Ecology and Restoration of Degraded Environments & School of Ecology and Environmental Sciences, Yunnan University, Kunming 650504, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2904-2063","authenticated-orcid":false,"given":"Xiao-Xia","family":"Pan","sequence":"additional","affiliation":[{"name":"School of Chemistry and Environment, Yunnan Minzu University, Kunming 650504, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3134-7643","authenticated-orcid":false,"given":"Ming-Zhi","family":"Yang","sequence":"additional","affiliation":[{"name":"Yunnan Key Laboratory for Plateau Mountain Ecology and Restoration of Degraded Environments & School of Ecology and Environmental Sciences, Yunnan University, Kunming 650504, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,8,12]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"58","DOI":"10.1186\/s40168-018-0445-0","article-title":"Microbial interactions within the plant holobiont","volume":"6","author":"Hassani","year":"2018","journal-title":"Microbiome"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.jare.2021.12.008","article-title":"The microbiota of the grapevine holobiont: A key component of plant health","volume":"40","author":"Bettenfeld","year":"2022","journal-title":"J. Adv. Res."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Boorboori, M.R., and Zhang, H.-Y. (2022). The role of Serendipita indica (Piriformospora indica) in improving plant resistance to drought and salinity stresses. Biology, 11.","DOI":"10.3390\/biology11070952"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Riseh, R.S., Fathi, F., Vazvani, M.G., and Tarkka, M.T. (2025). Plant colonization by biocontrol bacteria and improved plant health: A review. Front. Biosci.-Landmark, 30.","DOI":"10.31083\/FBL23223"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Raja, B.-L., Ait-El-Mokhtar, M., Abdelilah, M., and Marouane, B. (2026). Nodule\u2013microbiome dynamics: Deciphering the complexities of nodule symbiosis and the root microbiome. Int. J. Mol. Sci., 27.","DOI":"10.3390\/ijms27031487"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"fuaf063","DOI":"10.1093\/femsre\/fuaf063","article-title":"Cross-talk within plant niches: Endophytic and arbuscular mycorrhizal fungi for sustainable crop production","volume":"50","author":"Glick","year":"2026","journal-title":"FEMS Microbiol. Rev."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Chiquito-Contreras, C.J., Meza-Menchaca, T., Guzm\u00e1n-L\u00f3pez, O., V\u00e1squez, E.C., and Rica\u00f1o-Rodr\u00edguez, J. (2024). Molecular insights into plant\u2013microbe interactions: A comprehensive review of key mechanisms. Front. Biosci.-Elite, 16.","DOI":"10.31083\/j.fbe1601009"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"3567","DOI":"10.1038\/s41467-022-30971-8","article-title":"Host control and the evolution of cooperation in host microbiomes","volume":"13","author":"Sharp","year":"2022","journal-title":"Nat. Commun."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Chen, C.-X., Guo, L.-R., Wang, Y.-T., Wen, Y., Li, Y., Lu, C.-X., Zhou, P., Huang, S.-Y., Li, Y.-Q., and Pan, X.-X. (2024). Targeted Manipulation of Vertically Transmitted Endophytes to Confer Beneficial Traits in Grapevines. Horticulturae, 10.","DOI":"10.3390\/horticulturae10060607"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Tiwari, P., and Bae, H. (2020). Horizontal gene transfer and endophytes: An implication for the acquisition of novel traits. Plants, 9.","DOI":"10.3390\/plants9030305"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"513","DOI":"10.1016\/j.chom.2023.03.017","article-title":"Horizontal gene transfer among host-associated microbes","volume":"31","author":"Lourenco","year":"2023","journal-title":"Cell Host Microbe"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1111\/1758-2229.12181","article-title":"Bacterial seed endophytes: Genera, vertical transmission and interaction with plants","volume":"7","author":"Truyens","year":"2015","journal-title":"Environ. Microbiol. Rep."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"19255","DOI":"10.1021\/acs.jafc.3c07092","article-title":"Seed-to-seed: Plant core vertically transmitted microbiota","volume":"71","author":"Zeng","year":"2023","journal-title":"J. Agric. Food Chem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1038\/s41559-021-01593-y","article-title":"Natural selection for imprecise vertical transmission in host\u2013microbiota systems","volume":"6","author":"Bruijning","year":"2022","journal-title":"Nat. Ecol. Evol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"597","DOI":"10.1093\/icb\/icad031","article-title":"The microbiome as a maternal effect: A systematic review on vertical transmission of microbiota","volume":"63","author":"Murphy","year":"2023","journal-title":"Integr. Comp. Biol."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Sanz-Puente, I., Redondo-Salvo, S., Torres-Cort\u00e9s, G., de Toro, M., Fernandes, S., B\u00f6rner, A., Lorenzo, \u00d3., de la Cruz, F., and Robledo, M. (2025). Vertical transmission of core endophytes through the seeds. bioRxiv.","DOI":"10.1101\/2025.01.06.628327"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"wraf192","DOI":"10.1093\/ismejo\/wraf192","article-title":"Seed-mediated vertical transmission of Pantoea core endophytes","volume":"19","author":"Fernandes","year":"2025","journal-title":"ISME J."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Zhang, Y.-N., Hu, H.-Y., Li, Y., Geng, S.-C., Tang, J.-X., Pan, X.-X., and Yang, M.-Z. (2025). Identifying the Physiological Traits of Host-Dependent Endophytes in Grapevines, Using Callus as the Host Material. Microorganisms, 13.","DOI":"10.3390\/microorganisms13122791"},{"key":"ref_19","unstructured":"Lopez Fernandez, J. (2017). Molecular Ecological Interactions of Bacterial Endophytes with Their Host Vitis vinifera (L). [Ph.D. Thesis, Technische Universit\u00e4t Braunschweig]."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1705","DOI":"10.1039\/C7CS00664K","article-title":"Engineering chemical interactions in microbial communities","volume":"47","author":"Kenny","year":"2018","journal-title":"Chem. Soc. Rev."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"487","DOI":"10.1007\/s11240-016-1144-9","article-title":"Bacteria in the plant tissue culture environment","volume":"128","author":"Orlikowska","year":"2017","journal-title":"Plant Cell Tissue Organ Cult. (PCTOC)"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Cassells, A.C. (2012). Pathogen and biological contamination management in plant tissue culture: Phytopathogens, vitro pathogens, and vitro pests. Plant Cell Culture Protocols, Springer.","DOI":"10.1007\/978-1-61779-818-4_6"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"254","DOI":"10.37855\/jah.2025.v27i02.47","article-title":"Enhancing grapevine (Vitis vinifera L.) mass propagation: Single-node cuttings vs. traditional approaches","volume":"27","author":"Arab","year":"2025","journal-title":"J. Appl. Hortic."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Ozyigit, I.I., Dogan, I., Hocaoglu-Ozyigit, A., Yalcin, B., Erdogan, A., Yalcin, I.E., Cabi, E., and Kaya, Y. (2023). Production of secondary metabolites using tissue culture-based biotechnological applications. Front. Plant Sci., 14.","DOI":"10.3389\/fpls.2023.1132555"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2107","DOI":"10.1111\/1462-2920.14968","article-title":"The diversity and distribution of endophytes across biomes, plant phylogeny and host tissues: How far have we come and where do we go from here?","volume":"22","author":"Harrison","year":"2020","journal-title":"Environ. Microbiol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"983","DOI":"10.1071\/PP01101","article-title":"Niches for bacterial endophytes in crop plants: A plant biologist\u2019s view","volume":"28","author":"McCully","year":"2001","journal-title":"Aust. J. Plant Physiol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"381","DOI":"10.1007\/s11104-015-2495-4","article-title":"Endophytic bacterial community composition in wheat (Triticum aestivum) is determined by plant tissue type, developmental stage and soil nutrient availability","volume":"405","author":"Robinson","year":"2016","journal-title":"Plant Soil"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1186\/s13020-018-0198-5","article-title":"Diversity and composition of bacterial endophytes among plant parts of Panax notoginseng","volume":"13","author":"Dong","year":"2018","journal-title":"Chin. Med."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Babu, G.A., Mosa Christas, K., Kowsalya, E., Ramesh, M., Sohn, S.-I., and Pandian, S. (2022). Improved sterilization techniques for successful in vitro micropropagation. Commercial Scale Tissue Culture for Horticulture and Plantation Crops, Springer.","DOI":"10.1007\/978-981-19-0055-6_1"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"533","DOI":"10.1038\/ismej.2015.131","article-title":"Experimental insights into the importance of aquatic bacterial community composition to the degradation of dissolved organic matter","volume":"10","author":"Logue","year":"2016","journal-title":"ISME J."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Karlsson, I., Friberg, H., Steinberg, C., and Persson, P. (2014). Fungicide effects on fungal community composition in the wheat phyllosphere. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0111786"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"581","DOI":"10.1038\/nmeth.3869","article-title":"DADA2: High-resolution sample inference from Illumina amplicon data","volume":"13","author":"Callahan","year":"2016","journal-title":"Nat. Methods"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"852","DOI":"10.1038\/s41587-019-0209-9","article-title":"QIIME 2: Reproducible, interactive, scalable, and extensible microbiome data science","volume":"37","author":"Bolyen","year":"2019","journal-title":"Nat. Biotechnol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"685","DOI":"10.1038\/s41587-020-0548-6","article-title":"PICRUSt2 for prediction of metagenome functions","volume":"38","author":"Douglas","year":"2020","journal-title":"Nat. Biotechnol."},{"key":"ref_35","first-page":"133462","article-title":"BugBase predicts organism-level microbiome phenotypes","volume":"bioRxiv","author":"Ward","year":"2017","journal-title":"bioRxiv"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"241","DOI":"10.1016\/j.funeco.2015.06.006","article-title":"FUNGuild: An open annotation tool for parsing fungal community datasets by ecological guild","volume":"20","author":"Nguyen","year":"2016","journal-title":"Fungal Ecol."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Xiang, S.-Y., Wang, Y.-T., Chen, C.-X., Liao, C.-M., Li, T., Pan, X.-X., Zhu, S.-S., and Yang, M.-Z. (2023). Dominated \u201cinheritance\u201d of endophytes in grapevines from stock plants via in vitro-cultured plantlets: The dawn of plant endophytic modifications. Horticulturae, 9.","DOI":"10.3390\/horticulturae9020180"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1128\/MMBR.00050-14","article-title":"The hidden world within plants: Ecological and evolutionary considerations for defining functioning of microbial endophytes","volume":"79","author":"Hardoim","year":"2015","journal-title":"Microbiol. Mol. Biol. Rev."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"wrae073","DOI":"10.1093\/ismejo\/wrae073","article-title":"Mechanisms and implications of bacterial\u2013fungal competition for soil resources","volume":"18","author":"Wang","year":"2024","journal-title":"ISME J."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"5998","DOI":"10.1111\/1462-2920.16268","article-title":"Environmental micro-niche filtering shapes bacterial pioneer communities during primary colonization of a Himalayas\u2019 glacier forefield","volume":"24","author":"Rolli","year":"2022","journal-title":"Environ. Microbiol."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Martins, S., Brito, C., Baltazar, M., Dinis, L.-T., and Pereira, S. (2026). Exploring the Role of Root Exudates in Shaping Plant\u2013Soil\u2013Microbe Interactions to Support Agroecosystem Resilience. Horticulturae, 12.","DOI":"10.3390\/horticulturae12010090"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1111\/nph.70371","article-title":"Microbial drivers of root plasticity","volume":"248","author":"Wells","year":"2025","journal-title":"New Phytol."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Jones, P., Garcia, B.J., Furches, A., Tuskan, G.A., and Jacobson, D. (2019). Plant host-associated mechanisms for microbial selection. Front. Plant Sci., 10.","DOI":"10.3389\/fpls.2019.00862"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"416","DOI":"10.1007\/s00248-021-01688-w","article-title":"Plant compartments and developmental stages modulate the balance between niche-based and neutral processes in soybean microbiome","volume":"82","author":"Moroenyane","year":"2021","journal-title":"Microb. Ecol."},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Ait Si Mhand, K., Mouhib, S., Radouane, N., khatour, I., Aliyat, F.-Z., and Hijri, M. (2026). Compartment-Specific Assembly and Functional Potential of the Bacteriome of Citrullus colocynthis in a Semi-Arid Ecosystem. bioRxiv.","DOI":"10.64898\/2026.06.05.730417"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1016\/S1002-0160(19)60826-X","article-title":"A new perspective on functional redundancy and phylogenetic niche conservatism in soil microbial communities","volume":"30","author":"Yu","year":"2020","journal-title":"Pedosphere"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"936","DOI":"10.1038\/s41559-018-0519-1","article-title":"Function and functional redundancy in microbial systems","volume":"2","author":"Louca","year":"2018","journal-title":"Nat. Ecol. Evol."},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Zhang, C., Li, H., Li, M., Su, S., Xiao, H., Zhang, X., and Zhang, Q. (2025). Contrasting Assembly and Network Roles of Abundant and Rare Bacteria in Reservoir and Soil Habitats. Biology, 14.","DOI":"10.20944\/preprints202508.0509.v1"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"887","DOI":"10.1038\/s41477-022-01201-2","article-title":"Root-secreted bitter triterpene modulates the rhizosphere microbiota to improve plant fitness","volume":"8","author":"Zhong","year":"2022","journal-title":"Nat. Plants"},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Yuan, L., Jiang, H., Jiang, X., Li, T., Lu, P., Yin, X., and Wei, Y. (2022). Comparative genomic and functional analyses of Paenibacillus peoriae ZBSF16 with biocontrol potential against grapevine diseases, provide insights into its genes related to plant growth-promoting and biocontrol mechanisms. Front. Microbiol., 13.","DOI":"10.3389\/fmicb.2022.975344"}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/14\/8\/1780\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T11:33:33Z","timestamp":1787657613000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/14\/8\/1780"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,12]]},"references-count":50,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2026,8]]}},"alternative-id":["microorganisms14081780"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms14081780","relation":{},"ISSN":["2076-2607"],"issn-type":[{"value":"2076-2607","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,12]]}}}