Abstract
Main conclusion
The use of silver nanoparticles as elicitors in cell cultures of Rauwolfia serpentina resulted in increased levels of ajmalicine, upregulated structural and regulatory genes, elevated MDA content, and reduced activity of antioxidant enzymes. These findings hold potential for developing a cost-effective method for commercial ajmalicine production.
Abstract
Plants possess an intrinsic ability to detect various stress signals, prompting the activation of defense mechanisms through the reprogramming of metabolites to counter adverse conditions. The current study aims to propose an optimized bioprocess for enhancing the content of ajmalicine in Rauwolfia serpentina callus through elicitation with phytosynthesized silver nanoparticles. Initially, callus lines exhibiting elevated ajmalicine content were established. Following this, a protocol for the phytosynthesis of silver nanoparticles using seed extract from Rauwolfia serpentina was successfully standardized. The physicochemical attributes of the silver nanoparticles were identified, including their spherical shape, size ranging from 6.7 to 28.8 nm in diameter, and the presence of reducing-capping groups such as amino, carbonyl, and amide. Further, the findings indicated that the presence of 2.5 mg L-1 phytosynthesized silver nanoparticles in the culture medium increased the ajmalicine content. Concurrently, structural genes (TDC, SLS, STR, SGD, G10H) and regulatory gene (ORCA3) associated with the ajmalicine biosynthetic pathway were observed to be upregulated. A notable increase in MDA content and a decrease in the activities of antioxidant enzymes were observed. A notable increase in MDA content and a decrease in the activities of antioxidant enzymes were also observed. Our results strongly recommend the augmentation of ajmalicine content in the callus culture of R. serpentina through supplementation with silver nanoparticles, a potential avenue for developing a cost-effective process for the commercial production of ajmalicine.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
The data sets generated and/or analysed during the current study are available from the corresponding author on reasonable request.
References
Adejuwon AA, Oluwatoyin SM, Sunday AO (2011) Anti-inflammatory and antioxidant activities of hunteria umbellata seed fractions. Pharmacologia 2:165–171. https://doi.org/10.5567/pharmacologia.2011.165.171
Adeyemi OS, Sulaiman FA (2012) Biochemical and morphological changes in Trypanosoma brucei brucei-infected rats treated with homidium chloride and diminazene aceturate. J Basic Clin Physiol Pharmacol 23:179–183. https://doi.org/10.1515/jbcpp-2012-0018
Ali A, Mohammad S, Khan MA et al (2019) Silver nanoparticles elicited in vitro callus cultures for accumulation of biomass and secondary metabolites in Caralluma tuberculata. Artif Cells Nanomed Biotechnol 47:715–724. https://doi.org/10.1080/21691401.2019.1577884
Almagro L, Gutierrez J, Pedreño MA, Sottomayor M (2014) Synergistic and additive influence of cyclodextrins and methyl jasmonate on the expression of the terpenoid indole alkaloid pathway genes and metabolites in Catharanthus roseus cell cultures. Plant Cell Tissue Organ Cult 119:543–551. https://doi.org/10.1007/s11240-014-0554-9
Ambrin G, Ali HM, Ahmad A (2020) Metabolic regulation analysis of ajmalicine biosynthesis pathway in Catharanthus roseus (L.) G Don Suspension Culture using Nanosensor. Processes 8:589. https://doi.org/10.3390/PR8050589
Anjum S, Anjum I, Hano C, Kousar S (2019) Advances in nanomaterials as novel elicitors of pharmacologically active plant specialized metabolites: Current status and future outlooks. RSC Adv 9:40404–40423. https://doi.org/10.1039/c9ra08457f
Arrigoni O, de Gara L, Tommasi F, Liso R (1992) Changes in the ascorbate system during seed development of Vicia faba L. Plant Physiol 99:235–238. https://doi.org/10.1104/pp.99.1.235
Ashour AA, Raafat D, El-Gowelli HM, El-Kamel AH (2015) Green synthesis of silver nanoparticles using cranberry powder aqueous extract: characterization and antimicrobial properties. Int J Nanomed 10:7207–7221. https://doi.org/10.2147/IJN.S87268
Baghizadeh A, Ranjbar S, Gupta VK et al (2015) Green synthesis of silver nanoparticles using seed extract of Calendula officinalis in liquid phase. J Mol Liq 207:159–163. https://doi.org/10.1016/j.molliq.2015.03.029
Barbasz A, Kreczmer B, Oćwieja M (2016) Effects of exposure of callus cells of two wheat varieties to silver nanoparticles and silver salt (AgNO3). Acta Physiol Plant 38:1–11. https://doi.org/10.1007/s11738-016-2092-z
Beyer WF, Fridovich I (1987) Assaying for superoxide dismutase activity: Some large consequences of minor changes in conditions. Anal Biochem 161:559–566. https://doi.org/10.1016/0003-2697(87)90489-1
Bhagat P, Verma SK, Singh AK, et al (2019) Evaluation of influence of different strains of Agrobacterium rhizogenes on efficiency of hairy root induction in Rauwolfia serpentina. Indian J Genet Plant Breed 79:760–764. https://doi.org/10.31742/IJGPB.79.4.16
Bhagat P, Verma SK, Yadav S, et al (2020) Optimization of nutritive factors in culture medium for the growth of hairy root and ajmalicine content in sarpgandha, Rauwolfia serpentina. J Environ Biol 41:1018–1025. https://doi.org/10.22438/JEB/41/5/MRN-1316
Bindhu MR, Umadevi M (2013) Synthesis of monodispersed silver nanoparticles using Hibiscus cannabinus leaf extract and its antimicrobial activity. Spectrochim Acta A Mol Biomol Spectrosc 101:184–190. https://doi.org/10.1016/j.saa.2012.09.031
Bradford M (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254. https://doi.org/10.1006/abio.1976.9999
Cadet J, Wagner JR (2014) Oxidatively generated base damage to cellular DNA by hydroxyl radical and one-electron oxidants: Similarities and differences. Arch Biochem Biophys 557:47–54. https://doi.org/10.1016/j.abb.2014.05.001
Chmielowiec-Korzeniowska A, Tymczyna L, Dobrowolska M et al (2015) Silver (Ag) in tissues and eggshells, biochemical parameters and oxidative stress in chickens. Open Chem 13:1269–1274. https://doi.org/10.1515/chem-2015-0140
Chung IM, Rekha K, Rajakumar G, Thiruvengadam M (2018) Elicitation of silver nanoparticles enhanced the secondary metabolites and pharmacological activities in cell suspension cultures of bitter gourd. 3 Biotech 8:412. https://doi.org/10.1007/s13205-018-1439-0
Chung IM, Rekha K, Venkidasamy B, Thiruvengadam M (2019) Effect of copper oxide nanoparticles on the physiology, bioactive molecules, and transcriptional changes in brassica rapa ssp. rapa seedlings. Water Air Soil Pollut 230:1–14. https://doi.org/10.1007/s11270-019-4084-2
Cunha ADC, Chierrito TPC, MacHado GMDC et al (2012) Anti-leishmanial activity of alkaloidal extracts obtained from different organs of Aspidosperma ramiflorum. Phytomedicine 19:413–417. https://doi.org/10.1016/j.phymed.2011.12.004
Dey A, Roy D, Mohture VM et al (2022) Biotechnological interventions and indole alkaloid production in Rauvolfia serpentina. Appl Microbiol Biotechnol 106:4867–4883. https://doi.org/10.1007/s00253-022-12040-8
Dhindsa RS, Plumb-dhindsa P, Thorpe TA (1981) Leaf senescence: correlated with increased levels of membrane permeability and lipid peroxidation, and decreased levels of superoxide dismutase and catalase. J Exp Bot 32:93–101. https://doi.org/10.1093/jxb/32.1.93
Efferth T (2019) Biotechnology applications of plant callus cultures. Engineering 5:50–59. https://doi.org/10.1016/j.eng.2018.11.006
El-Sayed M, Verpoorte R (2007) Catharanthus terpenoid indole alkaloids: biosynthesis and regulation. Phytochem Rev 6:277–305. https://doi.org/10.1007/s11101-006-9047-8
El-Seedi HR, El-Shabasy RM, Khalifa SAM et al (2019) Metal nanoparticles fabricated by green chemistry using natural extracts: biosynthesis, mechanisms, and applications. RSC Adv 9:24539–24559. https://doi.org/10.1039/c9ra02225b
Farmer EE, Mueller MJ (2013) ROS-mediated lipid peroxidation and RES-activated signaling. Annu Rev Plant Biol 64:429–450. https://doi.org/10.1146/annurev-arplant-050312-120132
Fazaludeen Koya S, Pilakkadavath Z, Chandran P, et al (2022) Hypertension control rate in India: systematic review and meta-analysis of population-level non-interventional studies, 2001–2022
Ghorbanpour M, Hadian J (2015) Multi-walled carbon nanotubes stimulate callus induction, secondary metabolites biosynthesis and antioxidant capacity in medicinal plant Satureja khuzestanica grown in vitro. Carbon N Y 94:749–759. https://doi.org/10.1016/j.carbon.2015.07.056
Giri CC, Zaheer M (2016) Chemical elicitors versus secondary metabolite production in vitro using plant cell, tissue and organ cultures: recent trends and a sky eye view appraisal. Plant Cell Tissue Organ Cult 126:1–18. https://doi.org/10.1007/s11240-016-0985-6
Giri AK, Jena B, Biswal B et al (2022) Green synthesis and characterization of silver nanoparticles using Eugenia roxburghii DC. extract and activity against biofilm-producing bacteria. Sci Rep. https://doi.org/10.1038/s41598-022-12484-y
Goel MK, Mehrotra S, Kukreja AK, et al (2009) In vitro propagation of Rauwolfia serpentina using liquid medium, assessment of genetic fidelity of micropropagated plants, and simultaneous quantitation of reserpine, ajmaline, and ajmalicine. In: Methods in Molecular Biology. Humana Press Inc., pp 17–33
Goklany S, Rizvi NF, Loring RH et al (2013) Jasmonate-dependent alkaloid biosynthesis in Catharanthus Roseus hairy root cultures is correlated with the relative expression of Orca and Zct transcription factors. Biotechnol Prog 29:1367–1376. https://doi.org/10.1002/btpr.1801
Golkar P, Moradi M, Garousi GA (2019) Elicitation of stevia glycosides using salicylic acid and silver nanoparticles under callus culture. Sugar Tech 21:569–577. https://doi.org/10.1007/s12355-018-0655-6
He Y, Wei F, Ma Z et al (2017) Green synthesis of silver nanoparticles using seed extract of: Alpinia katsumadai, and their antioxidant, cytotoxicity, and antibacterial activities. RSC Adv 7:39842–39851. https://doi.org/10.1039/c7ra05286c
Ingold KU, Pratt DA (2014) Advances in radical-trapping antioxidant chemistry in the 21st century: a kinetics and mechanisms perspective. Chem Rev 114:9022–9046. https://doi.org/10.1021/cr500226n
Jagtap UB, Bapat VA (2013) Green synthesis of silver nanoparticles using Artocarpus heterophyllus Lam. seed extract and its antibacterial activity. Ind Crops Prod 46:132–137. https://doi.org/10.1016/j.indcrop.2013.01.019
Jamwal K, Bhattacharya S, Puri S (2018) Plant growth regulator mediated consequences of secondary metabolites in medicinal plants. J Appl Res Med Aromat Plants 9:26–38. https://doi.org/10.1016/J.JARMAP.2017.12.003
Karakaş Ö (2020) Effect of silver nanoparticles on production of indole alkaloids in Isatis constricta. Iran J Sci Technol Trans A Sci 44:621–627. https://doi.org/10.1007/s40995-020-00878-4
Kashyap P, Kalaiselvan V, Kumar R, Kumar S (2020) Ajmalicine and reserpine: Indole alkaloids as multi-target directed ligands towards factors implicated in Alzheimer’s disease. Molecules. https://doi.org/10.3390/molecules25071609
Khare N, Goyary D, Singh NK et al (2010) Transgenic tomato cv. Pusa Uphar expressing a bacterial mannitol-1-phosphate dehydrogenase gene confers abiotic stress tolerance. Plant Cell Tissue Organ Cult 103:267–277. https://doi.org/10.1007/s11240-010-9776-7
Khodakovskaya MV, de Silva K, Biris AS et al (2012) Carbon nanotubes induce growth enhancement of tobacco cells. ACS Nano 6:2128–2135. https://doi.org/10.1021/nn204643g
Kim DH, Gopal J, Sivanesan I (2017) Nanomaterials in plant tissue culture: the disclosed and undisclosed. RSC Adv 7:36492–36505. https://doi.org/10.1039/c7ra07025j
Kruszka D, Sawikowska A, Kamalabai Selvakesavan R et al (2020) Silver nanoparticles affect phenolic and phytoalexin composition of Arabidopsis thaliana. Sci Total Environ 716:135361. https://doi.org/10.1016/J.SCITOTENV.2019.135361
Lawal B, Shittu O, Ossai P et al (2015) Evaluation of antioxidant activity of giant African snail (Achachatina maginata) haemolymph in CCl4- induced hepatotoxixity in albino rats. Br J Pharm Res 6:141–154. https://doi.org/10.9734/bjpr/2015/15887
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods 25:402–408. https://doi.org/10.1006/meth.2001.1262
Luczkiewicz M, Kokotkiewicz A, Glod D (2014) Plant growth regulators affect biosynthesis and accumulation profile of isoflavone phytoestrogens in high-productive in vitro cultures of Genista tinctoria. Plant Cell Tissue Organ Cult 118:419–429. https://doi.org/10.1007/s11240-014-0494-4
Mahajan S, Kadam J, Dhawal P, et al (2022) Application of silver nanoparticles in in-vitro plant growth and metabolite production: revisiting its scope and feasibility. Plant Cell Tissue Organ Culture (PCTOC) 2022 150:1 150:15–39. https://doi.org/10.1007/S11240-022-02249-W
Mahjouri S, Movafeghi A, Divband B, Kosari-Nasab M (2018) Toxicity impacts of chemically and biologically synthesized CuO nanoparticles on cell suspension cultures of Nicotiana tabacum. Plant Cell Tissue Organ Cult 135:223–234. https://doi.org/10.1007/s11240-018-1458-x
Marslin G, Sheeba CJ, Franklin G (2017) Nanoparticles alter secondary metabolism in plants via ROS burst. Front Plant Sci 8:832. https://doi.org/10.3389/fpls.2017.00832
Menke FLH, Champion A, Kijne JW, Memelink J (1999) A novel jasmonate- and elicitor-responsive element in the periwinkle secondary metabolite biosynthetic gene Str interacts with a jasmonate- and elicitor-inducible AP2-domain transcription factor, ORCA2. EMBO J 18:4455–4463. https://doi.org/10.1093/emboj/18.16.4455
Meraj TA, Fu J, Raza MA et al (2020) Transcriptional factors regulate plant stress responses through mediating secondary metabolism. Genes (basel) 11:346. https://doi.org/10.3390/genes11040346
Miyake C, Shinzaki Y, Nishioka M et al (2006) Photoinactivation of ascorbate peroxidase in isolated tobacco chloroplasts: Galdieria partita APX maintains the electron flux through the water-water cycle in transplastomic tobacco plants. Plant Cell Physiol 47:200–210. https://doi.org/10.1093/pcp/pci235
Nabikhan A, Kandasamy K, Raj A, Alikunhi NM (2010) Synthesis of antimicrobial silver nanoparticles by callus and leaf extracts from saltmarsh plant, Sesuvium portulacastrum L. Colloids Surf B Biointerfaces 79:488–493. https://doi.org/10.1016/j.colsurfb.2010.05.018
Narayani M, Srivastava S (2017) Elicitation: a stimulation of stress in in vitro plant cell/tissue cultures for enhancement of secondary metabolite production. Phytochem Rev 16:1227–1252. https://doi.org/10.1007/s11101-017-9534-0
Naveed M, Bukhari B, Aziz T, et al (2022) Green synthesis of silver nanoparticles using the plant extract of acer oblongifolium and study of its antibacterial and antiproliferative activity via mathematical approaches. molecules 27:4226 27:4226. https://doi.org/10.3390/MOLECULES27134226
Nayyar H, Gupta D (2006) Differential sensitivity of C3 and C4 plants to water deficit stress: association with oxidative stress and antioxidants. Environ Exp Bot 58:106–113. https://doi.org/10.1016/j.envexpbot.2005.06.021
Pan Q, Wang Q, Yuan F et al (2012) Overexpression of ORCA3 and G10H in Catharanthus roseus plants regulated alkaloid biosynthesis and metabolism revealed by NMR-metabolomics. PLoS ONE 7:e43038. https://doi.org/10.1371/journal.pone.0043038
Pan YJ, Liu J, Guo XR et al (2015) Gene transcript profiles of the TIA biosynthetic pathway in response to ethylene and copper reveal their interactive role in modulating TIA biosynthesis in Catharanthus roseus. Protoplasma 252:813–824. https://doi.org/10.1007/s00709-014-0718-9
Park JS, Seong ZK, Kim MS et al (2020) Production of flavonoids in callus cultures of Sophora flavescens aiton. Plants. https://doi.org/10.3390/plants9060688
Pimprikar PS, Joshi SS, Kumar AR et al (2009) Influence of biomass and gold salt concentration on nanoparticle synthesis by the tropical marine yeast Yarrowia lipolytica NCIM 3589. Colloids Surf B Biointerfaces 74:309–316. https://doi.org/10.1016/j.colsurfb.2009.07.040
Poborilova Z, Opatrilova R, Babula P (2013) Toxicity of aluminium oxide nanoparticles demonstrated using a BY-2 plant cell suspension culture model. Environ Exp Bot 91:1–11. https://doi.org/10.1016/J.ENVEXPBOT.2013.03.002
Qidwai A, Kumar R, Dikshit A (2018) Green synthesis of silver nanoparticles by seed of Phoenix sylvestris L. and their role in the management of cosmetics embarrassment. Green Chem Lett Rev 11:176–188. https://doi.org/10.1080/17518253.2018.1445301
Ramani S, Patil N, Nimbalkar S, Jayabaskaran C (2013) Alkaloids derived from tryptophan: Terpenoid indole alkaloids. In: Natural Products: Phytochemistry, Botany and Metabolism of Alkaloids, Phenolics and Terpenes. Springer Berlin Heidelberg, pp 575–604
Rani A, Guleria M, Sharma Y et al (2023) Insights into elicitor’s role in augmenting secondary metabolites production and climate resilience in genus Ocimum – A globally important medicinal and aromatic crop. Ind Crops Prod 202:117078. https://doi.org/10.1016/j.indcrop.2023.117078
Sangodele JO, Olaleye MT, Monsees TK, Akinmoladun AC (2017) The para isomer of dinitrobenzene disrupts redox homeostasis in liver and kidney of male wistar rats. Biochem Biophys Rep 10:297–302. https://doi.org/10.1016/j.bbrep.2017.04.017
Sathishkumar M, Sneha K, Won SW et al (2009) Cinnamon zeylanicum bark extract and powder mediated green synthesis of nano-crystalline silver particles and its bactericidal activity. Colloids Surf B Biointerfaces 73:332–338. https://doi.org/10.1016/j.colsurfb.2009.06.005
Schmittgen TD, Livak KJ (2008) Analyzing real-time PCR data by the comparative CT method. Nat Protoc 3:1101–1108. https://doi.org/10.1038/nprot.2008.73
Sharma A, Mathur AK, Ganpathy J et al (2019) Effect of abiotic elicitation and pathway precursors feeding over terpenoid indole alkaloids production in multiple shoot and callus cultures of Catharanthus roseus. Biologia (bratisl) 74:543–553. https://doi.org/10.2478/s11756-019-00202-5
Shasmita BS, Mishra P et al (2023) Recent advances in tissue culture and secondary metabolite production in Hypericum perforatum L. Plant Cell Tissue Organ Cult 154:13–28. https://doi.org/10.1007/S11240-023-02525-3/METRICS
Singh S, Pandey SS, Shanker K, Kalra A (2020) Endophytes enhance the production of root alkaloids ajmalicine and serpentine by modulating the terpenoid indole alkaloid pathway in Catharanthus roseus roots. J Appl Microbiol 128:1128–1142. https://doi.org/10.1111/jam.14546
Srivastava M, Singh S, Self WT (2012) Exposure to silver nanoparticles inhibits selenoprotein synthesis and the activity of thioredoxin reductase. Environ Health Perspect 120:56–61. https://doi.org/10.1289/ehp.1103928
Suriyakalaa U, Antony JJ, Suganya S et al (2013) Hepatocurative activity of biosynthesized silver nanoparticles fabricated using Andrographis paniculata. Colloids Surf B Biointerfaces 102:189–194. https://doi.org/10.1016/j.colsurfb.2012.06.039
Tiwari S, Verma SK, Bhagat P et al (2021) An overview of the phytosynthesis of various metal nanoparticles. 3 Biotech 11:1–10. https://doi.org/10.1007/S13205-021-03014-0
Vidyasagar N, Patel RR, Singh SK, Singh M (2023) Green synthesis of silver nanoparticles: methods, biological applications, delivery and toxicity. Mater Adv 4:1831–1849
Waseem M, Naveed M, Rehman S, ur, et al (2023) Molecular characterization of spa, hld, fmhA, and lukD genes and computational modeling the multidrug resistance of staphylococcus species through Callindra harrisii silver nanoparticles. ACS Omega 8:20920–20936. https://doi.org/10.1021/acsomega.3c01597
Yang Z, Xiao Y, Jiao T et al (2020) Effects of copper oxide nanoparticles on the growth of rice (Oryza sativa L.) seedlings and the relevant physiological responses. Int J Environ Res Public Health 17:1260. https://doi.org/10.3390/ijerph17041260
Yasur J, Rani PU (2013) Environmental effects of nanosilver: Impact on castor seed germination, seedling growth, and plant physiology. Environ Sci Pollut Res 20:8636–8648. https://doi.org/10.1007/s11356-013-1798-3
Zahir A, Nadeem M, Ahmad W et al (2019) Chemogenic silver nanoparticles enhance lignans and neolignans in cell suspension cultures of Linum usitatissimum L. Plant Cell Tissue Organ Cult 136:589–596. https://doi.org/10.1007/s11240-018-01539-6
Zhang B, Zheng LP, Yi Li W, Wen Wang J (2013) Stimulation of artemisinin production in Artemisia annua hairy roots by Ag-SiO2 core-shell nanoparticles. Curr Nanosci 9:363–370. https://doi.org/10.2174/1573413711309030012
Zhang H, Du W, Peralta-Videa JR et al (2018) Metabolomics reveals how cucumber (Cucumis sativus) reprograms metabolites to cope with silver ions and silver nanoparticle-induced oxidative stress. Environ Sci Technol 52:8016–8026. https://doi.org/10.1021/acs.est.8b02440
Zhou P, Yang J, Zhu J et al (2015) Effects of β-cyclodextrin and methyl jasmonate on the production of vindoline, catharanthine, and ajmalicine in Catharanthus roseus cambial meristematic cell cultures. Appl Microbiol Biotechnol 99:7035–7045. https://doi.org/10.1007/s00253-015-6651-9
Zia F, Ghafoor N, Iqbal M, Mehboob S (2016) Green synthesis and characterization of silver nanoparticles using Cydonia oblong seed extract. Appl Nanosci (Switzerland) 6:1023–1029. https://doi.org/10.1007/s13204-016-0517-z
Funding
We are grateful to Science and Engineering Research Board, Department of Science and Technology, India (Grant number SB/YS/LS-96/2014) for financial support to Neeraj Khare. We are also grateful to SERB-DST for financial support in the form of Fellowship of Sachin Kumar Verma.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Additional information
Communicated by De-Yu Xie.
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Verma, S.K., Goyary, D., Singh, A.K. et al. Modulation of terpenoid indole alkaloid pathway via elicitation with phytosynthesized silver nanoparticles for the enhancement of ajmalicine, a pharmaceutically important alkaloid. Planta 259, 30 (2024). https://doi.org/10.1007/s00425-023-04311-z
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s00425-023-04311-z