{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,3]],"date-time":"2026-06-03T14:22:09Z","timestamp":1780496529582,"version":"3.54.1"},"reference-count":37,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2023,12,28]],"date-time":"2023-12-28T00:00:00Z","timestamp":1703721600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Shenzhen Science and Technology Program from The Science, Technology, and Innovation Commission of Shenzhen Municipality","award":["JCYJ20210324123005016"],"award-info":[{"award-number":["JCYJ20210324123005016"]}]},{"name":"Shenzhen Science and Technology Program from The Science, Technology, and Innovation Commission of Shenzhen Municipality","award":["JCYJ20230807151000001"],"award-info":[{"award-number":["JCYJ20230807151000001"]}]},{"name":"Shenzhen Science and Technology Program from The Science, Technology, and Innovation Commission of Shenzhen Municipality","award":["KCXFZ20211020164200001"],"award-info":[{"award-number":["KCXFZ20211020164200001"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The sesquiterpene alkaloid dendrobine, widely recognized as the main active compound and a quality control standard of medicinal orchids in the Chinese Pharmacopoeia, demonstrates diverse biological functions. In this study, we engineered Dendrobium catenatum as a chassis plant for the production of dendrobine through the screening and pyramiding of key biosynthesis genes. Initially, previously predicted upstream key genes in the methyl-D-erythritol 4-phosphate (MEP) pathway for dendrobine synthesis, including 4-(Cytidine 5\u2032-Diphospho)-2-C-Methyl-d-Erythritol Kinase (CMK), 1-Deoxy-d-Xylulose 5-Phosphate Reductoisomerase (DXR), 2-C-Methyl-d-Erythritol 4-Phosphate Cytidylyltransferase (MCT), and Strictosidine Synthase 1 (STR1), and a few downstream post-modification genes, including Cytochrome P450 94C1 (CYP94C1), Branched-Chain-Amino-Acid Aminotransferase 2 (BCAT2), and Methyltransferase-like Protein 23 (METTL23), were chosen due to their deduced roles in enhancing dendrobine production. The seven genes (SG) were then stacked and transiently expressed in the leaves of D. catenatum, resulting in a dendrobine yield that was two-fold higher compared to that of the empty vector control (EV). Further, RNA-seq analysis identified Copper Methylamine Oxidase (CMEAO) as a strong candidate with predicted functions in the post-modification processes of alkaloid biosynthesis. Overexpression of CMEAO increased dendrobine content by two-fold. Additionally, co-expression analysis of the differentially expressed genes (DEGs) by weighted gene co-expression network analysis (WGCNA) retrieved one regulatory transcription factor gene MYB61. Overexpression of MYB61 increased dendrobine levels by more than two-fold in D. catenatum. In short, this work provides an efficient strategy and prospective candidates for the genetic engineering of D. catenatum to produce dendrobine, thereby improving its medicinal value.<\/jats:p>","DOI":"10.3390\/ijms25010397","type":"journal-article","created":{"date-parts":[[2023,12,28]],"date-time":"2023-12-28T03:30:34Z","timestamp":1703734234000},"page":"397","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Metabolic Pathway Engineering Improves Dendrobine Production in Dendrobium catenatum"],"prefix":"10.3390","volume":"25","author":[{"given":"Meili","family":"Zhao","sequence":"first","affiliation":[{"name":"Shenzhen Key Laboratory for Orchid Conservation and Utilization, The National Orchid Conservation Center of China and the Orchid Conservation & Research Center of Shenzhen, Shenzhen 518114, China"},{"name":"Key Laboratory of National Forestry and Grassland Administration for Orchid Conservation and Utilization, The National Orchid Conservation Center of China and the Orchid Conservation & Research Center of Shenzhen, Shenzhen 518114, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanchang","family":"Zhao","sequence":"additional","affiliation":[{"name":"State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, South China Agricultural University, Guangzhou 510642, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhenyu","family":"Yang","sequence":"additional","affiliation":[{"name":"Shenzhen Key Laboratory for Orchid Conservation and Utilization, The National Orchid Conservation Center of China and the Orchid Conservation & Research Center of Shenzhen, Shenzhen 518114, China"},{"name":"Shanghai Key Laboratory of Plant Molecular Sciences, College of Life Sciences, Shanghai Normal University, Shanghai 200234, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1788-6218","authenticated-orcid":false,"given":"Feng","family":"Ming","sequence":"additional","affiliation":[{"name":"Shanghai Key Laboratory of Plant Molecular Sciences, College of Life Sciences, Shanghai Normal University, Shanghai 200234, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jian","family":"Li","sequence":"additional","affiliation":[{"name":"Shenzhen Key Laboratory for Orchid Conservation and Utilization, The National Orchid Conservation Center of China and the Orchid Conservation & Research Center of Shenzhen, Shenzhen 518114, China"},{"name":"Key Laboratory of National Forestry and Grassland Administration for Orchid Conservation and Utilization, The National Orchid Conservation Center of China and the Orchid Conservation & Research Center of Shenzhen, Shenzhen 518114, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Demin","family":"Kong","sequence":"additional","affiliation":[{"name":"Shenzhen Key Laboratory for Orchid Conservation and Utilization, The National Orchid Conservation Center of China and the Orchid Conservation & Research Center of Shenzhen, Shenzhen 518114, China"},{"name":"Key Laboratory of National Forestry and Grassland Administration for Orchid Conservation and Utilization, The National Orchid Conservation Center of China and the Orchid Conservation & Research Center of Shenzhen, Shenzhen 518114, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu","family":"Wang","sequence":"additional","affiliation":[{"name":"Shenzhen Key Laboratory for Orchid Conservation and Utilization, The National Orchid Conservation Center of China and the Orchid Conservation & Research Center of Shenzhen, Shenzhen 518114, China"},{"name":"Key Laboratory of National Forestry and Grassland Administration for Orchid Conservation and Utilization, The National Orchid Conservation Center of China and the Orchid Conservation & Research Center of Shenzhen, Shenzhen 518114, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peng","family":"Chen","sequence":"additional","affiliation":[{"name":"Shenzhen Key Laboratory for Orchid Conservation and Utilization, The National Orchid Conservation Center of China and the Orchid Conservation & Research Center of Shenzhen, Shenzhen 518114, China"},{"name":"Key Laboratory of National Forestry and Grassland Administration for Orchid Conservation and Utilization, The National Orchid Conservation Center of China and the Orchid Conservation & Research Center of Shenzhen, Shenzhen 518114, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Meina","family":"Wang","sequence":"additional","affiliation":[{"name":"Shenzhen Key Laboratory for Orchid Conservation and Utilization, The National Orchid Conservation Center of China and the Orchid Conservation & Research Center of Shenzhen, Shenzhen 518114, China"},{"name":"Key Laboratory of National Forestry and Grassland Administration for Orchid Conservation and Utilization, The National Orchid Conservation Center of China and the Orchid Conservation & Research Center of Shenzhen, Shenzhen 518114, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhicai","family":"Wang","sequence":"additional","affiliation":[{"name":"Shenzhen Key Laboratory for Orchid Conservation and Utilization, The National Orchid Conservation Center of China and the Orchid Conservation & Research Center of Shenzhen, Shenzhen 518114, China"},{"name":"Key Laboratory of National Forestry and Grassland Administration for Orchid Conservation and Utilization, The National Orchid Conservation Center of China and the Orchid Conservation & Research Center of Shenzhen, Shenzhen 518114, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,12,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"e9107","DOI":"10.7717\/peerj.9107","article-title":"Comparative metabolomic analyses of Dendrobium officinale Kimura et Migo responding to UV-B radiation reveal variations in the metabolisms associated with its bioactive ingredients","volume":"8","author":"Chen","year":"2020","journal-title":"PeerJ"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"2080","DOI":"10.1016\/j.apsb.2021.01.019","article-title":"The chromosome-level reference genome assembly for dendrobium officinale and its utility of functional genomics research and molecular breeding study","volume":"11","author":"Niu","year":"2021","journal-title":"Acta Pharm. Sin. B."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"808228","DOI":"10.3389\/fpls.2021.808228","article-title":"Research advances in multi-omics on the traditional Chinese herb Dendrobium officinale","volume":"12","author":"Wang","year":"2021","journal-title":"Front. Plant Sci."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1016\/j.plaphy.2022.08.026","article-title":"Transcriptomic and metabolomic analyses reveal the main metabolites in Dendrobium officinale leaves during the harvesting period","volume":"190","author":"Si","year":"2022","journal-title":"Plant Physiol. Biochem."},{"key":"ref_5","first-page":"874","article-title":"Protocorm-like bodies (PLBs) of Dendrobium Sabin Blue: A novel source for in vitro production of dendrobine and anthocyanin","volume":"57","author":"Chin","year":"2021","journal-title":"In Vitro Cell Dev. Biol."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Cao, H., Ji, Y.L., Li, S.H., Lu, L., Tian, M., Yang, W., and Li, H. (2019). Extensive metabolic profiles of leaves stems from the medicinal plant Dendrobium officinale Kimura et Migo. Metabolites, 9.","DOI":"10.3390\/metabo9100215"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"419","DOI":"10.1007\/s10265-019-01099-6","article-title":"Comparative transcriptomic analysis reveal the regulation mechanism underlying MeJA-induced accumulation of alkaloids in Dendrobium officinale","volume":"132","author":"Chen","year":"2019","journal-title":"J. Plant Res."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Liu, J.J., Liu, Z.P., Zhang, X.F., and Si, J.P. (2021). Effects of various processing methods on the metabolic profile and antioxidant activity of Dendrobium catenatum Lindley leaves. Metabolites, 11.","DOI":"10.3390\/metabo11060351"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"154132","DOI":"10.1016\/j.phymed.2022.154132","article-title":"Alkaloids from Dendrobium and their biosynthetic pathway, biological activity and total synthesis","volume":"102","author":"Duan","year":"2022","journal-title":"Phytomedicine"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"379","DOI":"10.1111\/bcpt.13524","article-title":"Dendrobine attenuates gestational diabetes mellitus in mice by inhibiting Th17 cells","volume":"128","author":"Feng","year":"2021","journal-title":"Basic. Clin. Pharmacol. Toxicol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"6597","DOI":"10.1007\/s00253-021-11534-1","article-title":"Recent advances and new insights in biosynthesis of dendrobine and sesquiterpenes","volume":"105","author":"Gong","year":"2021","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Jia, Q., Wang, L., Qian, X., Jin, H., Shu, F., Sarsaiya, S., Jin, L., and Chen, J. (2022). Transcriptome analysis of dendrobine biosynthesis in Trichoderma longibrachiatum MD33. Front. Microbiol., 13.","DOI":"10.3389\/fmicb.2022.890733"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"104208","DOI":"10.1016\/j.arabjc.2022.104208","article-title":"UPLC-Q\/TOF-MS coupled with multivariate analysis for comparative analysis of metabolomic in Dendrobium nobile from different growth altitudes","volume":"15","author":"Lu","year":"2022","journal-title":"Arab. J. Chem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"850949","DOI":"10.3389\/fpls.2022.850949","article-title":"Natural composition and biosynthetic pathways of alkaloids in medicinal Dendrobium species","volume":"13","author":"Song","year":"2022","journal-title":"Front. Plant Sci."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"129","DOI":"10.1186\/s13568-022-01470-2","article-title":"Functional characterization of a farnesyl diphosphate synthase from Dendrobium nobile Lindl","volume":"12","author":"Gong","year":"2022","journal-title":"AMB Express"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Jiao, C., Song, C., Zheng, S., Zhu, Y., Jin, Q., Cai, Y., and Lin, Y. (2018). Metabolic profiling of Dendrobium officinale in response to precursors and methyl jasmonate. Int. J. Mol. Sci., 19.","DOI":"10.3390\/ijms19030728"},{"key":"ref_17","first-page":"1347","article-title":"Effects of different drying methods on polysaccharides and dendrobine from Dendrobium candidum","volume":"44","author":"Xin","year":"2013","journal-title":"J. South. Agric."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"19029","DOI":"10.1038\/srep19029","article-title":"The Dendrobium catenatum Lindl. genome sequence provides insights into polysaccharide synthase, floral development and adaptive evolution","volume":"6","author":"Zhang","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"391","DOI":"10.3389\/fpls.2020.00391","article-title":"Transcriptomic landscape of medicinal Dendrobium reveals genes associated with the biosynthesis of bioactive components","volume":"11","author":"Wang","year":"2020","journal-title":"Front. Plant Sci."},{"key":"ref_20","first-page":"1751","article-title":"Phytohormone-triggered transcriptional changes revealed \u03b2-glucosidase as a key player for polysaccharide metabolism in Dendrobium officinale","volume":"49","author":"Wang","year":"2022","journal-title":"Prog. Biochem. Biophys."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Wang, Z., Zhao, M., Zhang, X., Deng, X., Li, J., and Wang, M. (2022). Genome-wide identification and characterization of active ingredients related \u03b2-glucosidases in Dendrobium catenatum. BMC Genom., 23.","DOI":"10.1186\/s12864-022-08840-x"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"769","DOI":"10.1007\/s00425-018-2960-4","article-title":"Genome-wide researches and applications on Dendrobium","volume":"248","author":"Zheng","year":"2018","journal-title":"Planta"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"316","DOI":"10.1038\/s41598-017-00445-9","article-title":"Transcriptome analysis of genes involved in dendrobine biosynthesis in Dendrobium nobile Lindl. infected with mycorrhizal fungus MF23 (Mycena sp.)","volume":"7","author":"Li","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"e13664","DOI":"10.7554\/eLife.13664","article-title":"A new synthetic biology approach allows transfer of an entire metabolic pathway from a medicinal plant to a biomass crop","volume":"5","author":"Fuentes","year":"2016","journal-title":"eLife"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"115375","DOI":"10.1016\/j.indcrop.2022.115375","article-title":"The Dendrobium catenatum DcCIPK24 increases drought and salt tolerance of transgenic Arabidopsis","volume":"187","author":"Zhang","year":"2022","journal-title":"Ind. Crops Prod."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Zhang, M., Yu, Z., Zeng, D., Si, C., Zhao, C., Wang, H., Li, C., He, C., and Duan, J. (2021). Transcriptome and metabolome reveal salt-stress responses of leaf tissues from Dendrobium officinale. Biomolecules, 11.","DOI":"10.3390\/biom11050736"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Yuan, Y., Yu, M., Jia, Z., Song Xe Liang, Y., and Zhang, J. (2018). Analysis of Dendrobium huoshanense transcriptome unveils putative genes associated with active ingredients synthesis. BMC Genom., 19.","DOI":"10.1186\/s12864-018-5305-6"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Wang, Z., Jiang, W., Liu, Y., Meng, X., Su, X., Cao, M., Wu, L., Yu, N., Xing, S., and Peng, D. (2021). Putative genes in alkaloid biosynthesis identified in Dendrobium officinale by correlating the contents of major bioactive metabolites with genes expression between protocorm-like bodies and leaves. BMC Genom., 22.","DOI":"10.1186\/s12864-021-07887-6"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"17700","DOI":"10.1038\/s41598-020-74789-0","article-title":"Metabolomic and transcriptomic analyses reveal the regulation of pigmentation in the purple variety of Dendrobium officinale","volume":"10","author":"Zhan","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"520","DOI":"10.1111\/tpj.13509","article-title":"AaMYB1 and its orthologue AtMYB61 affect terpene metabolism and trichome development in Artemisia annua and Arabidopsis thaliana","volume":"90","author":"Jiang","year":"2017","journal-title":"Plant J."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1194","DOI":"10.1016\/j.molp.2020.06.009","article-title":"TBtools: An integrative toolkit developed for interactive analyses of big biological data","volume":"13","author":"Chen","year":"2020","journal-title":"Mol. Plant"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Langfelder, P., and Horvath, S. (2008). WGCNA: An R package for weighted correlation network analysis. BMC Bioinform., 9.","DOI":"10.1186\/1471-2105-9-559"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"100471","DOI":"10.1016\/j.xplc.2022.100471","article-title":"A Nimble Cloning-compatible vector system for high-throughput gene functional analysis in plants","volume":"4","author":"Yan","year":"2023","journal-title":"Plant Commun."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1983","DOI":"10.1111\/pbi.13882","article-title":"Efficient assembly of long DNA fragments and multiple genes with improved nickase-based cloning and Cre\/loxP recombination","volume":"20","author":"Zhao","year":"2022","journal-title":"Plant Biotechnol. J."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"641","DOI":"10.1038\/nprot.2006.97","article-title":"Agrobacterium-mediated transformation of Arabidopsis thaliana using the floral dip method","volume":"1","author":"Zhang","year":"2006","journal-title":"Nat. Protoc."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"2071","DOI":"10.1111\/jipb.13534","article-title":"ERF5 enhances protocorm-like body regeneration via enhancement of STM expression in Dendrobium orchid","volume":"65","author":"Zeng","year":"2023","journal-title":"J. Integr. Plant Biol."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Lau, C., Siskos, A.P., Maitre, L., Robinson, O., Athersuch, T.J., Want, E.J., Urquiza, J., Casas, M., Vafeiadi, M., and Roumeliotaki, T. (2018). Determinants of the urinary and serum metabolome in children from six European populations. BMC Med., 16.","DOI":"10.1186\/s12916-018-1190-8"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/1\/397\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T21:43:06Z","timestamp":1760132586000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/1\/397"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,12,28]]},"references-count":37,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2024,1]]}},"alternative-id":["ijms25010397"],"URL":"https:\/\/doi.org\/10.3390\/ijms25010397","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,12,28]]}}}