In vivo Metabolic Investigation of Oxygen, Light, and Temperature Effects on Dormancy Alleviation of Sesame (Sesamum indicum L.) Seeds

Year 2025, Volume: 31 Issue: 1, 196 - 206, 14.01.2025

Honghao Cai , Xiayi Ruan Yumin Wan Mengting Chen Xianqin Wu Yingqiang Cai

Abstract

As an edible seed, sesame seeds require careful storage to maintain their quality. Dormancy helps seeds extend their lifespan by slowing down metabolic processes, reducing energy consumption and natural aging. However, seeds may exit dormancy and begin germination during storage due to variations in temperature, light, and oxygen conditions. This transition is not easily visible, but nutritional components within the seeds can start to deplete. In this study, non-invasive magnetic resonance spectroscopy and imaging were used to monitor sesame seeds stored under different temperature, light, and oxygen conditions for over 120 hours. Results showed that seeds remained dormant at 15 °C under oxygen deprivation and in the absence of light. When exposed to continuous light at 15 °C, under anaerobic or aerobic conditions, changes in metabolic resonances were observed through spectroscopy, indicating moisture and fatty acid transfer between seed structures. Despite these changes, magnetic resonance imaging showed that the embryo did not develop. At 24 °C with continuous light and aerobic conditions, both spectroscopy and imaging analyses revealed significant metabolic changes, and all internal seed structures developed normally, with visible signs of germination. This study highlights that although sesame seeds are non-photoblastic, light can still trigger metabolic activity within the seeds, while suitable temperature is essential for complete seed development. These findings provide valuable insights into the dynamic molecular-level metabolic changes from dormancy to early seed germination using magnetic resonance technology and offer guidance for maintaining seed dormancy during storage.

Keywords

NMR, MRI, 1H NMR spectroscopy, Chemical Shift Selective Imaging, Germination, Storage

Ethical Statement

We, the authors of this manuscript, hereby declare that the research work presented in this paper was conducted ethically and responsibly. The following ethical principles have been adhered to in the preparation of this manuscript: Originality and Plagiarism: We affirm that the content of this manuscript is original and has not been plagiarized from other sources. All sources used in this research have been appropriately cited and referenced. Authorship and Contribution: All individuals who have made significant contributions to the research and preparation of this manuscript are listed as co-authors. Additionally, all authors have reviewed and approved the final version of the manuscript before submission. Data Integrity: We confirm that all data presented in this manuscript are accurate and have not been manipulated or fabricated. The raw data is available and can be provided upon request for verification purposes. Conflict of Interest: All potential conflicts of interest, financial or otherwise, that could be perceived as influencing the research outcomes have been disclosed. The authors declare no competing interests in relation to this work. Ethical Approval: If applicable, the research described in this manuscript has been approved by the relevant institutional review boards or ethics committees. All procedures performed in studies involving human participants or animals were in accordance with the ethical standards of the institution and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. Informed Consent: In studies involving human participants, informed consent was obtained from all individual participants included in the study. The privacy rights of human subjects have been observed, and personal data is kept confidential.

Supporting Institution

Jimei University

References

• Allen D K, Ohlrogge J B & Shachar‐Hill Y (2009). The role of light in soybean seed filling metabolism. The Plant Journal 58 (2): 220-234. https://doi.org/10.1111/j.1365313X.2008.03771.x.
• Amanah H Z, Joshi R, Masithoh R E, Choung M G, Kim K H, Kim G & Cho B K (2020). Nondestructive measurement of anthocyanin in intact soybean seed using Fourier Transform Near-Infrared (FT-NIR) and Fourier Transform Infrared (FT-IR) spectroscopy. Infrared Physics and Technology 111: 103477. https://doi.org/10.1016/j.infrared.2020.103477.
• Bharti S K & Roy R (2012). Quantitative 1H NMR spectroscopy. TrAC Trends in Analytical Chemistry 35: 5-26. https://doi.org/10.1016/j.trac.2012.02.007.
• Blystone S, Nuixe M, Traoré A S, Cochard H, Picon-Cochard C & Pagés G (2024). Towards portable MRI in the plant sciences. Plant Methods 20 (1): 31. https://doi.org/10.1186/s13007-024-01152-z.
• Boulc’h P N, Collewet G, Guillon B, Quellec S, Leport L & Musse M (2024). Quantitative MRI imaging of parenchyma and venation networks in Brassica napus leaves: effects of development and dehydration. Plant Methods 20(1): 69. https://doi.org/10.1186/s13007-024-01187-2.
• Cai H, Cheng R, Jin Y, Ding S & Chen Z (2016a). Evaluation of oolong teas using 1H and 13C solid‐state NMR, sensory analysis, and multivariate Statistics. Journal of the Chinese Chemical Society 63(9): 792-799. https://doi.org/10.1002/jccs.201600183.
• Cai H, Lin L, Ding S, Cui X & Chen Z (2016b). Fast quantification of fatty acid profile of intact fish by intermolecular double‐quantum coherence 1H‐NMR spectroscopy. European Journal of Lipid Science and Technology 118(6): 1150-1159. https://doi.org/10.1002/ejlt.201500309.
• Cai H, Chuang W, Cui X, Cheng R, Chiu K, Chen Z & Ding S (2018a). High resolution 31P NMR spectroscopy generates a quantitative evolution profile of phosphorous translocation in germinating sesame seed. Scientific Reports 8: 359. https://doi.org/10.1038/s41598-017-18722-y.
• Cai H, Jin Y & Cui X (2018b). Feasibility of ultrafast high‐resolution spectroscopy in the analysis of molecular‐mobility‐restricted samples in deuterium‐free environments. Journal of the Chinese Chemical Society 65(6): 674-680. https://doi.org/10.1002/jccs.201700430.
• Cao J, Li G, Qu D, Li X & Wang Y (2020). Into the seed: auxin controls seed development and grain yield. International Journal of Molecular Sciences 21 (5): 1662. https://doi.org/10.3390/ijms21051662.
• Caprara C, Mathias T K, Santos M, D’Oca M G, D’Oca C D R, Roselet F, Abreu P C & Ramos D F (2023). Application of 1H HR-MAS NMR-based metabolite fingerprinting of marine microalgae. Metabolites 13(2): 202. https://doi.org/10.3390/metabo13020202.
• Carosio M G, Bernardes D F, Carvalho A & Colnago L A (2018). Non-invasive measurements of oilseed temperature in soil and soil thermal diffusivity using time-domain NMR relaxometry. Applied Magnetic Resonance 49(10): 1119-1127. https://doi.org/10.1007/s00723-018-1028-8.
• Castroverde C D M & Dina D (2021). Temperature regulation of plant hormone signaling during stress and development. Journal of Experimental Botany 72(21): 7436-7458. https://doi.org/10.1093/jxb/erab257.
• Chahtane H, Kim W & Lopez-Molina L (2017). Primary seed dormancy: A temporally multilayered riddle waiting to be unlocked. Journal of Experimental Botany 68(4): 857-869. https://doi.org/10.1093/jxb/erw377.
• Chen D, Yuan H, Bao J, Zhao X, Fu X & Hu X (2023). Dry storage alters intraspecific variation in phenotypic traits at early life stages: evidence from a dominant alpine meadow species. Seed Science Research 33(2): 203-212. https://doi.org/10.1017/S0960258523000223.
• Cheng Y, Chen H, Zhao Y, Cheng X, Wang L & Guo X (2023). Effect of light quality on polyphenol biosynthesis in three varieties of mung bean sprouts with different color seed coats. Plant Cell Reports 42(2): 253-268. https://doi.org/10.1007/s00299-022-02954-y.
• Chiwocha S D, Abrams S R, Ambrose S J, Cutler A J, Loewen M, Ross A R & Kermode A R (2003). A method for profiling classes of plant hormones and their metabolites using liquid chromatography‐electrospray ionization tandem mass spectrometry: an analysis of hormone regulation of thermos dormancy of lettuce (Lactuca sativa L.) seeds. The Plant Journal 35(3): 405-417. https://doi.org/10.1046/j.1365-313X.2003.01800.x.
• Corbineau F (2022) Oxygen, a key signaling factor in the control of seed germination and dormancy. Seed Science Research 32(3): 126-136. https://doi.org/10.1017/S096025852200006X.
• d'Avignon D A & Ge X (2018). In vivo NMR investigations of glyphosate influences on plant metabolism. Journal of Magnetic Resonance 292: 59-72. https://doi.org/10.1016/j.jmr.2018.03.008.
• Dossa K, Diouf D, Wang L, Wei X, Zhang Y, Niang M, Fonceka D, Yu J, Mmadi M A & Yehouessi L W (2017). The emerging oilseed crop Sesamum indicum enters the “Omics” era. Frontiers in Plant Science 8: 1154. https://doi.org/10.3389/fpls.2017.01154.
• Egli D & Wardlaw I (1980). Temperature response of seed growth characteristics of soybeans. Agronomy Journal 72(3): 560-564. https://doi.org/10.2134/agronj1980.00021962007200030036x.
• Elboghdady A E A, Gomma A H, Hamed A M & Abdallatif A M (2023). Assessment of phenotypic diversity of some date palm male genotypes growing under Egyptian conditions. Revista Brasileira De Fruticultura 45: e-896. https://doi.org/10.1590/0100-29452023896.
• Fujii H, Sato N, Kimura Y, Mizutani M, Kusama M, Sumitomo N, Chiba E, Shigemoto Y, Takao M & Takayama Y (2022). MR imaging detection of CNS lesions in tuberous sclerosis complex: The usefulness of T1WI with chemical shift selective images. American Journal of Neuroradiology 43(8): 1202-1209. https://doi.org/10.3174/ajnr.A7573.
• Gao Z, Chen S, Huang J & Cai H (2024a). Real‐time quantitative detection of hydrocolloid adulteration in meat based on Swin Transformer and smartphone. Journal of Food Science 89(7): 4359-4371. https://doi.org/10.1111/1750-3841.17159.
• Gao Z, Huang J, Chen J, Shao T, Ni H & Cai H (2024b). Deep transfer learning-based computer vision for real-time harvest period classification and impurity detection of Porphyra haitnensis. Aquaculture International 32: 5171-5198. https://doi.org/10.1007/s10499-024-01422-6.
• Gay C, Corbineau F & Côme D (1991). Effects of temperature and oxygen on seed germination and seedling growth in sunflower (Helianthus annuus L.). Environmental and Experimental Botany 31(2): 193-200. 10.1016/0098-8472(91)90070-5.
• Gebeyehu B (2020). Review on: Effect of seed storage period and storage environment on seed quality. International Journal of Applied Agricultural Sciences 6(6): 185-190. https://doi.org/10.11648/J.IJAAS.20200606.14.
• Geneve R L (2003). Impact of temperature on seed dormancy. HortScience 38(3): 336-340. https://doi.org/10.21273/HORTSCI.38.3.336.
• Haase A, Frahm J, Hanicke W & Matthaei D (1985). 1H NMR chemical shift selective (CHESS) imaging. Physics in Medicine and Biology 30(4): 341. https://doi.org/10.1088/0031-9155/30/4/008.
• Jander G, Norris S R, Joshi V, Fraga M, Rugg A, Yu S, Li L & Last R L (2004). Application of a high‐throughput HPLC‐MS/MS assay to Arabidopsis mutant screening; evidence that threonine aldolase plays a role in seed nutritional quality. The Plant Journal 39(3): 465-475. https://doi.org/10.1111/j.1365-313X.2004.02140.x.
• Kauth P J & Biber P D (2015). Moisture content, temperature, and relative humidity influence seed storage and subsequent survival and germination of Vallisneria americana seeds. Aquatic Botany 120: 297-303. https://doi.org/10.1016/j.aquabot.2014.09.009.
• Kim C S (1983). Studies on the germination characteristics of sesame (Sesamum indicum L.). Korean Journal of Agricultural Science 10(1): 28-60
• Kołodziejczyk I, Bałabusta M, Szewczyk R & Posmyk M M (2015). The levels of melatonin and its metabolites in conditioned corn (Zea mays L.) and cucumber (Cucumis sativus L.) seeds during storage. Acta Physiologiae Plantarum 37: 1-11. https://doi.org/10.1007/s11738-015-1850-7.
• Liao W, Cai H & Ni H (2024). Sesame seed metabolism during germination under auxin: An in vivo NMR study. Journal of Plant Growth Regulation. https://doi.org/10.1007/s00344-024-11574-7
• Li G, Chen M, Chen J, Shang Y, Lian X, Wang P, Lei H & Ma Q (2020). Chemical composition analysis of pomegranate seeds based on ultra-high-performance liquid chromatography coupled with quadrupole-Orbitrap high-resolution mass spectrometry. Journal of Pharmaceutical and Biomedical Analysis 187: 113357. https://doi.org/10.1016/j.jpba.2020.113357.
• Li X, Simpson W, Song M, Bao G, Niu X, Zhang Z, Xu H, Liu X, Li Y & Li C (2020). Effects of seed moisture content and Epichloe endophyte on germination and physiology of Achnatherum inebrians. South African Journal of Botany 134: 407-414. https://doi.org/10.1016/j.sajb.2020.03.022.
• Ma T, Tsuchikawa S & Inagaki T (2020). Rapid and non-destructive seed viability prediction using near-infrared hyperspectral imaging coupled with a deep learning approach. Computers and Electronics in Agriculture 177: 105683. https://doi.org/10.1016/j.compag.2020.105683.
• Mota M F S, Waktola H D, Nolvachai Y & Marriott P J (2021). Gas chromatography‒mass spectrometry for characterisation, assessment of quality and authentication of seed and vegetable oils. TrAC Trends in Analytical Chemistry 138: 116238. https://doi.org/10.1016/j.trac.2021.116238.
• Munz E, Rolletschek H, Oeltze‐Jafra S, Fuchs J, Guendel A, Neuberger T, Ortleb S, Jakob P M & Borisjuk L (2017). A functional imaging study of germinating oilseed rape seed. New Phytologist 216: 1181-1190. https://doi.org/10.1111/nph.14736.
• Pasternak T, Pérez-Pérez J M, Ruperti B, Aleksandrova T & Palme K (2024). A new in vitro growth system for phenotypic characterization and seed propagation of arabidopsis thaliana. Journal of Plant Growth Regulation 43(2): 652-658. https://doi.org/10.1007/s00344-023-11093-x.
• Qiu J, Lin Y, Wu J, Xiao Y, Cai H & Ni H (2024). Rapid beef quality detection using spectra pre‐processing methods in electrical impedance spectroscopy and machine learning. International Journal of Food Science and Technology 59(3): 1624-1634. https://doi.org/10.1111/ijfs.16915.
• Sarkar B K, Yang W, Wu Z, Tang H & Ding S (2009). Variations of water uptake, lipid consumption, and dynamics during the germination of Sesamum indicum seed: A nuclear magnetic resonance spectroscopic investigation. Journal of Agricultural and Food Chemistry 57(18): 8213-8219. https://doi.org/10.1021/jf9019129.
• Simon E, Minchin A, McMenamin M M & Smith J (1976). The low temperature limit for seed germination. New Phytologist 77(2): 301-311. https://doi.org/10.1111/j.1469-8137.1976.tb01519.x.
• Song L, Wang Q, Wang C, Lin Y, Yu D, Xu Z, Huang Q & Wu Y (2015). Effect of γ-irradiation on rice seed vigor assessed by near-infrared spectroscopy. Journal of Stored Products Research 62: 46-51. https://doi.org/10.1016/j.jspr.2015.03.009.
• Tang W, Wu N, Xiao Q, Chen S, Gao P, He Y & Feng L (2023). Early detection of cotton verticillium wilt based on root magnetic resonance images. Frontiers in Plant Science 14: 1135718. https://doi.org/10.3389/fpls.2023.1135718.
• Taylor A, Lee P & Zhang M (1999). Volatile compounds as indicators of seed quality and their influence on seed aging. Seed Technology 21 (1): 57-65.
• Terskikh V, Müller K, Kermode A R & Leubner-Metzger G (2011). In vivo 1H-NMR microimaging during seed imbibition, germination, and early growth. Seed Dormancy: Methods and Protocols 773: 319-327. https://doi.org/10.1007/978-1-61779-231-1_18.
• Tigabu M & Odén P C (2004). Rapid and non-destructive analysis of vigour of Pinus patula seeds using single seed near infrared transmittance spectra and multivariate analysis. Seed Science and Technology 32(2): 593-606. https://doi.org/10.15258/sst.2004.32.2.28.
• Tuomainen T V, Toljamo A, Kokko H & Nissi M J (2024). Non-invasive assessment and visualization of Phytophthora cactorum infection in strawberry crowns using quantitative magnetic resonance imaging. Scientific Reports 14(1): 2129. https://doi.org/10.1038/s41598-024-52520-7.
• Wu H, Deng J, Yang C, Zhang J, Zhang Q, Wang X, Yang F, Yang W & Liu J (2017). Metabolite profiling of isoflavones and anthocyanins in black soybean [Glycine max (L.) Merr.] seeds by HPLC-MS and geographical differentiation analysis in Southwest China. Analytical Methods 9(5): 792-802. https://doi.org/10.1039/C6AY02970A.
• Xiao Y, Cai H & Ni H (2024). Identification of geographical origin and adulteration of Northeast China soybeans by mid-infrared spectroscopy and spectra augmentation. Journal of Consumer Protection and Food Safety 19(1): 99-111. https://doi.org/10.1007/s00003-023-01471-8.
• Yan A & Chen Z (2020). The control of seed dormancy and germination by temperature, light and nitrate. The Botanical Review 86: 39-75. https://doi.org/10.1007/s12229-020-09220-4.
• Xiao Y, Liao W, Cai H & Ni H (2024). Dynamic investigation of auxin on germination of sesamum indicum seed by 1H spectroscopy and chemical shift selection Imaging. Chiang Mai Journal of Science 51: e2024033. https://doi.org/10.12982/CMJS.2024.033.
• Yan S, Huang W, Gao J, Fu H & Liu J (2018). Comparative metabolomic analysis of seed metabolites associated with seed storability in rice (Oryza sativa L.) during natural aging. Plant Physiology and Biochemistry 127: 590-598. https://doi.org/10.1016/j.plaphy.2018.04.020.
• Yin L Y, Zhang R F, Xie Z M, Wang C Y & Li W (2013). The effect of temperature, substrate, light, oxygen availability and burial depth on Ottelia alismoides seed germination. Aquatic Botany 111: 50-53. https://doi.org/10.1016/j.aquabot.2013.09.001.
• Zhang M, Zhu J & Yan Q (2012). Review on influence mechanisms of light in seed germination. Chinese Journal of Plant Ecology 36(8): 899. https://doi.org/10.3724/SP.J.1258.2012.00899.
• Zhang T, Fan S, Xiang Y, Zhang S, Wang J & Sun Q (2020). Non-destructive analysis of germination percentage, germination energy and simple vigour index on wheat seeds during storage by Vis/NIR and SWIR hyperspectral imaging. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 239: 118488. https://doi.org/10.1016/j.saa.2020.118488.