{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T09:10:07Z","timestamp":1789031407153,"version":"build-2803163510"},"reference-count":41,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T00:00:00Z","timestamp":1788998400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100013064","name":"Key Research and Development Program of Jiangxi Province","doi-asserted-by":"publisher","award":["20261BCF320042"],"award-info":[{"award-number":["20261BCF320042"]}],"id":[{"id":"10.13039\/501100013064","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Biological Breeding-Major Projects","award":["2023ZD04058"],"award-info":[{"award-number":["2023ZD04058"]}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32160389"],"award-info":[{"award-number":["32160389"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"award":["32160389"],"award-info":[{"award-number":["32160389"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Jiangxi Province Early Career Youth Science and Technology Talent Training Project","award":["20262BEJ730222"],"award-info":[{"award-number":["20262BEJ730222"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Plants"],"abstract":"<jats:p>Somatic embryogenesis (SE) is a core technology for the clonal propagation of forest tree species. However, the low frequency of normal embryos and high rate of abnormal embryos during SE maturation induction in slash pine (Pinus elliottii) severely constrain its commercial application. In this study, comparative transcriptome analysis between normal embryos (NE) and abnormal embryos (AE) revealed that glutathione metabolism and riboflavin metabolism pathways may be involved in the regulation of somatic embryo maturation. Based on these findings, different concentration gradients of reduced glutathione (GSH) and riboflavin were supplemented into the somatic embryo maturation medium to verify their effects. Single-factor experiments showed that the 0.5 mM GSH treatment yielded 158.33 normal embryos per gram of embryogenic callus with a normal embryo ratio of 42.20%, while the 7.5 \u00b5M riboflavin treatment yielded 100 normal embryos per gram with a normal embryo ratio of 18.27%. Further 2 \u00d7 2 orthogonal combination experiments demonstrated that the synergistic treatment of 0.5 mM GSH and 7.5 \u00b5M riboflavin achieved the optimal effect, producing 333.33 normal embryos per gram of embryogenic callus with a normal embryo ratio of 72.88%. Both maturation rate and induction efficiency were significantly superior to single treatments and other combinations. These results demonstrate that GSH and riboflavin can synergistically improve the normal embryo ratio and accelerate somatic embryo maturation in slash pine. The underlying mechanisms involving metabolic and hormonal pathways warrant further physiological and biochemical investigation. This study provides a technical reference for optimizing the somatic embryogenesis system of slash pine.<\/jats:p>","DOI":"10.3390\/plants15182768","type":"journal-article","created":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T08:07:41Z","timestamp":1789027661000},"page":"2768","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Synergistic Promotion of Somatic Embryogenesis in Pinus elliottii by Reduced Glutathione and Riboflavin"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-6422-6345","authenticated-orcid":false,"given":"Lin","family":"Xu","sequence":"first","affiliation":[{"name":"Jiangxi Academy of Forestry, Nanchang 330013, China"},{"name":"Jiangxi Provincial Key Laboratory of Improved Variety Breeding and Efficient Utilization of Native Tree Species, Nanchang 330013, China"},{"name":"Key Laboratory of National Forestry and Grassland Administration on Forest Cultivation and Management in East China, Nanchang 330013, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"visualization","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"}]},{"given":"Wei","family":"Ding","sequence":"additional","affiliation":[{"name":"Jiangxi Academy of Forestry, Nanchang 330013, China"},{"name":"Jiangxi Provincial Key Laboratory of Improved Variety Breeding and Efficient Utilization of Native Tree Species, Nanchang 330013, China"},{"name":"Key Laboratory of National Forestry and Grassland Administration on Forest Cultivation and Management in East China, Nanchang 330013, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"}]},{"given":"Xi","family":"Ouyang","sequence":"additional","affiliation":[{"name":"Jiangxi Academy of Forestry, Nanchang 330013, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"formal-analysis","vocabulary":"credit"}]},{"given":"Bin","family":"Gong","sequence":"additional","affiliation":[{"name":"Jiangxi Academy of Forestry, Nanchang 330013, China"},{"name":"Jiangxi Provincial Key Laboratory of Improved Variety Breeding and Efficient Utilization of Native Tree Species, Nanchang 330013, China"},{"name":"Key Laboratory of National Forestry and Grassland Administration on Forest Cultivation and Management in East China, Nanchang 330013, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"resources","vocabulary":"credit"}]},{"given":"Qian","family":"Liu","sequence":"additional","affiliation":[{"name":"Jiangxi Academy of Forestry, Nanchang 330013, China"},{"name":"Jiangxi Provincial Key Laboratory of Improved Variety Breeding and Efficient Utilization of Native Tree Species, Nanchang 330013, China"},{"name":"Key Laboratory of National Forestry and Grassland Administration on Forest Cultivation and Management in East China, Nanchang 330013, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"investigation","vocabulary":"credit"}]},{"given":"Zhaolei","family":"Deng","sequence":"additional","affiliation":[{"name":"Jiangxi Academy of Forestry, Nanchang 330013, China"},{"name":"Jiangxi Provincial Key Laboratory of Improved Variety Breeding and Efficient Utilization of Native Tree Species, Nanchang 330013, China"},{"name":"Key Laboratory of National Forestry and Grassland Administration on Forest Cultivation and Management in East China, Nanchang 330013, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"resources","vocabulary":"credit"}]},{"given":"Chunxia","family":"Yang","sequence":"additional","affiliation":[{"name":"Jiangxi Academy of Forestry, Nanchang 330013, China"},{"name":"Jiangxi Provincial Key Laboratory of Improved Variety Breeding and Efficient Utilization of Native Tree Species, Nanchang 330013, China"},{"name":"Key Laboratory of National Forestry and Grassland Administration on Forest Cultivation and Management in East China, Nanchang 330013, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"supervision","vocabulary":"credit"},{"role":"project-administration","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"100867","DOI":"10.1016\/j.tfp.2025.100867","article-title":"Forest tree breeding under the global environmental change: Challenges and opportunities","volume":"20","author":"Li","year":"2025","journal-title":"Trees For. People"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"44","DOI":"10.1007\/s11240-025-03172-6","article-title":"Mapping fifty-five years of somatic embryogenesis research: A bibliometric analysis of trends, global collaborations, and key advances across more than nine thousand articles","volume":"162","year":"2025","journal-title":"Plant Cell Tissue Organ Cult. (PCTOC)"},{"key":"ref_3","first-page":"e010","article-title":"Genetic transformation in conifers: Current status and future prospects","volume":"4","author":"Zhao","year":"2024","journal-title":"For. Res."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1007\/s00425-025-04849-0","article-title":"Application and development prospect of genomic selection breeding in coniferous trees","volume":"262","author":"Zhang","year":"2025","journal-title":"Planta"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"259","DOI":"10.1007\/s42977-022-00126-3","article-title":"Genetic and epigenetic modes of the regulation of somatic embryogenesis: A review","volume":"73","author":"Sivanesan","year":"2022","journal-title":"Biol. Futur."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Xu, L., Deng, Z., Hu, S., Liu, Q., Luan, Q., and Yang, C. (2025). Establishment of efficient somatic embryo maturation system of Pinus elliottii. Plants, 14.","DOI":"10.3390\/plants14131985"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Salaj, T., Klubicov\u00e1, K., Matusova, R., and Salaj, J. (2019). Somatic embryogenesis in selected conifer trees Pinus nigra Arn. and Abies Hybrids. Front. Plant Sci., 10.","DOI":"10.3389\/fpls.2019.00013"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1079\/IVP2001262","article-title":"Maturation of somatic embryos in conifers: Morphogenesis, physiology, biochemistry, and molecular biology","volume":"38","author":"Stasolla","year":"2002","journal-title":"Vitr. Cell. Dev. Biol.-Plant"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"874","DOI":"10.1093\/plcell\/koac337","article-title":"Endosperm cellularization failure induces a dehydration-stress response leading to embryo arrest","volume":"35","author":"Xu","year":"2023","journal-title":"Plant Cell"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"104436","DOI":"10.1016\/j.envexpbot.2021.104436","article-title":"ABA-induced stomatal movements in vascular plants during dehydration and rehydration","volume":"186","author":"Hasan","year":"2021","journal-title":"Environ. Exp. Bot."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"527","DOI":"10.1007\/s11240-022-02301-9","article-title":"PEG induces maturation of somatic embryos of Passiflora edulis Sims \u2018UENF Rio Dourado\u2019 by differential accumulation of proteins and modulation of endogenous contents of free polyamines","volume":"150","author":"Cruz","year":"2022","journal-title":"Plant Cell Tissue Organ Cult. (PCTOC)"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"184","DOI":"10.1007\/s44371-026-00642-4","article-title":"Antimicrobial and detoxification potentials of plant-based activated carbon: Current advances and applications","volume":"3","author":"Dalhatu","year":"2026","journal-title":"Discov. Chem."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1002\/reg2.91","article-title":"A transcriptional view on somatic embryogenesis","volume":"4","author":"Horstman","year":"2017","journal-title":"Regeneration"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.mam.2008.08.006","article-title":"Glutathione: Overview of its protective roles, measurement, and biosynthesis","volume":"30","author":"Forman","year":"2009","journal-title":"Mol. Asp. Med."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"4549","DOI":"10.1093\/jxb\/erae194","article-title":"Glutathione: A key modulator of plant defence and metabolism through multiple mechanisms","volume":"75","author":"Noctor","year":"2024","journal-title":"J. Exp. Bot."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"4078","DOI":"10.1038\/s41467-025-59255-7","article-title":"Structure and transport mechanism of human riboflavin transporters","volume":"16","author":"Wang","year":"2025","journal-title":"Nat. Commun."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"418","DOI":"10.1016\/j.freeradbiomed.2024.09.002","article-title":"Maternal riboflavin deficiency causes embryonic defects by activating ER stress-induced hepatocyte apoptosis pathway","volume":"224","author":"Zhang","year":"2024","journal-title":"Free Radic. Biol. Med."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Georgiou-Siafis, S.K., and Tsiftsoglou, A.S. (2023). The key role of GSH in keeping the redox balance in mammalian cells: Mechanisms and significance of GSH in detoxification via formation of Conjugates. Antioxidants, 12.","DOI":"10.3390\/antiox12111953"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Iyanagi, T. (2022). Roles of ferredoxin-NADP+ oxidoreductase and flavodoxin in NAD(P)H-dependent electron transfer systems. Antioxidants, 11.","DOI":"10.3390\/antiox11112143"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"6418","DOI":"10.1093\/jxb\/erab306","article-title":"ABA signalling promotes cell totipotency in the shoot apex of germinating embryos","volume":"72","author":"Chen","year":"2021","journal-title":"J. Exp. Bot."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2253","DOI":"10.1105\/tpc.109.067702","article-title":"Abscisic acid represses growth of the Arabidopsis embryonic axis after germination by enhancing auxin signaling","volume":"21","author":"Belin","year":"2009","journal-title":"Plant Cell"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"108969","DOI":"10.1016\/j.plaphy.2024.108969","article-title":"ABA exerts a promotive effect on the early process of somatic embryogenesis in Quercus aliena Bl","volume":"214","author":"Yang","year":"2024","journal-title":"Plant Physiol. Biochem."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Chen, Y., Fei, Q., Xia, X., Ke, X., Ye, J., and Zhu, L. (2023). Pinus massoniana somatic embryo maturation, mycorrhization of regenerated plantlets and its resistance to Bursaphelenchus xylophilus. Front. Plant Sci., 14.","DOI":"10.3389\/fpls.2023.1130471"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"356","DOI":"10.1111\/j.1469-8137.2007.02289.x","article-title":"Inhibited polar auxin transport results in aberrant embryo development in Norway spruce","volume":"177","author":"Larsson","year":"2008","journal-title":"New Phytol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"648","DOI":"10.1038\/s41580-025-00851-2","article-title":"Mechanisms of auxin action in plant growth and development","volume":"26","author":"Vanneste","year":"2025","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"154394","DOI":"10.1016\/j.jplph.2024.154394","article-title":"Alteration in the balance between ABA and GA signaling mediates genetic variation in induction and retention of dormancy during seed maturation in wheat","volume":"304","author":"Nguyen","year":"2025","journal-title":"J. Plant Physiol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1039","DOI":"10.1016\/j.cj.2020.11.006","article-title":"Brassinosteroid and gibberellin coordinate rice seed germination and embryo growth by regulating glutelin mobilization","volume":"9","author":"Xiong","year":"2021","journal-title":"Crop J."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1154","DOI":"10.1016\/j.freeradbiomed.2015.09.023","article-title":"Glutathione\u2014Linking cell proliferation to oxidative stress","volume":"89","author":"Kiddle","year":"2015","journal-title":"Free Radic. Biol. Med."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"123721","DOI":"10.1016\/j.indcrop.2026.123721","article-title":"Glutathione metabolism maintains redox homeostasis to enhance somatic embryogenesis in hybrid larch","volume":"249","author":"Zhu","year":"2026","journal-title":"Ind. Crops Prod."},{"key":"ref_30","first-page":"5","article-title":"Glutathione promotes somatic embryogenesis by regulating ThGPX4 in Taxodium hybrid \u2018zhongshanshan\u2019","volume":"43","author":"Chen","year":"2026","journal-title":"Tree Physiol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"2218","DOI":"10.1038\/s41598-023-28387-5","article-title":"Reactive oxygen metabolism in the proliferation of Korean pine embryogenic callus cells promoted by exogenous GSH","volume":"13","author":"Gao","year":"2023","journal-title":"Sci. Rep."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1466","DOI":"10.1007\/s00344-023-11198-3","article-title":"Effects of Exogenous 2,4-Dichlorophenoxyacetic Acid, Glutathione, and L-Buthionine Sulfoximine on Intracellular Redox State and Hormone Levels and Their Links with Proliferation in Picea pungens Somatic Embryogenesis","volume":"43","author":"Gao","year":"2024","journal-title":"J. Plant Growth Regul."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1799","DOI":"10.1093\/plphys\/kiad139","article-title":"Riboflavin mediates m6A modification targeted by miR408, promoting early somatic embryogenesis in longan","volume":"192","author":"Xu","year":"2023","journal-title":"Plant Physiol."},{"key":"ref_34","first-page":"638","article-title":"Progress in the Research on Riboflavin Biosynthesis and Function in Plants","volume":"58","author":"Hu","year":"2023","journal-title":"Chin. Bull. Bot."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"32","DOI":"10.1016\/j.gene.2019.04.041","article-title":"Thioredoxins and thioredoxin reductase in chloroplasts: A review","volume":"706","author":"Kang","year":"2019","journal-title":"Gene"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1569","DOI":"10.1111\/nph.16086","article-title":"Arabidopsis glutathione reductase 2 is indispensable in plastids, while mitochondrial glutathione is safeguarded by additional reduction and transport systems","volume":"224","author":"Marty","year":"2019","journal-title":"New Phytol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"3405","DOI":"10.1093\/jxb\/eraa107","article-title":"The pivotal function of dehydroascorbate reductase in glutathione homeostasis in plants","volume":"71","author":"Ding","year":"2020","journal-title":"J. Exp. Bot."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Cao, X., Yang, H., Shang, C., Ma, S., Liu, L., and Cheng, J. (2019). The roles of auxin biosynthesis YUCCA gene family in plants. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20246343"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"5173","DOI":"10.1093\/jxb\/erz268","article-title":"Maize Dek33 encodes a pyrimidine reductase in riboflavin biosynthesis that is essential for oil-body formation and ABA biosynthesis during seed development","volume":"70","author":"Dai","year":"2019","journal-title":"J. Exp. Bot."},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Ruan, J., Zhou, Y., Zhou, M., Yan, J., Khurshid, M., Weng, W., Cheng, J., and Zhang, K. (2019). Jasmonic acid signaling pathway in plants. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20102479"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"241","DOI":"10.1007\/s10265-003-0096-4","article-title":"Structure and function of cytokinin oxidase\/dehydrogenase genes of maize, rice, Arabidopsis and other species","volume":"116","author":"Werner","year":"2003","journal-title":"J. Plant Res."}],"container-title":["Plants"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2223-7747\/15\/18\/2768\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T08:42:09Z","timestamp":1789029729000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2223-7747\/15\/18\/2768"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,10]]},"references-count":41,"journal-issue":{"issue":"18","published-online":{"date-parts":[[2026,9]]}},"alternative-id":["plants15182768"],"URL":"https:\/\/doi.org\/10.3390\/plants15182768","relation":{},"ISSN":["2223-7747"],"issn-type":[{"value":"2223-7747","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,10]]}}}