Abstract
Salvia viridis, cultivated in Europe as an ornamental plant, has been used for centuries in folk medicine in its natural habitat in the areas of Iran and Turkey. The aim of the study was to obtain a S. viridis shoot culture that can provide high amounts of raw material rich in bioactive compounds at the lowest possible cost in a short time. It investigated the effect of different light spectra (including blue, red, mixed i.e. R:B 7:3 and white LEDs, and fluorescent lamps), and different liquid media systems (liquid agitated medium, liquid stationary medium without support, and with nanocellulose or polyurethane as support) on culture growth and polyphenol production. The phenolic compounds in 80% methanolic extract were quantified by ultra-high performance liquid chromatography (UPLC). It was found that among the LED treatments, the blue light had the most beneficial effect on shoot regeneration (proliferation factor of 4.56), shoot biomass (17-fold increase in fresh weight, 15-fold increase in dry weight), and polyphenol level (15.24 mg/g dry weight). Additionally, a liquid system with nanocellulose as a supporting material turned out to be beneficial for the cultivation of S. viridis. In these conditions, about 8.4 new buds/shoots were obtained from a single explant, fresh weight increased about 40-fold, and dry weight 30-fold. The liquid culture system with cellulose yielded 38.5 mg/g DW polyphenol content: twice as high as the shoots on the solid medium.
Key message
Liquid system with nanocellulose and blue LEDs result in efficient biomass and total polyphenol accumulation in Salvia viridis shoots.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
All data generated or analysed during this study are included in this article.
References
Bach A, Kapczyńska A, Dziurka K, Dziurka M (2018) The importance of applied light quality on the process of shoot organogenesis and production of phenolics and carbohydrates in Lachenalia sp. cultures in vitro. S Afr J Bot 114:14–19. https://doi.org/10.1016/j.sajb.2017.10.015
Batista DS, Felipe SHS, Silva TD, de Castro KM, Mamedes-Rodrigues TC, Miranda NA, Ríos-Ríos AM, Faria DV, Fortini EA, Chagas K, Torres-Silva G, Xavier A, Arencibia AD, Otoni WC (2018) Light quality in plant tissue culture: does it matter? In Vitro Cell Dev Biol Plant 54:195–215. https://doi.org/10.1007/s11627-018-9902-5
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248-254B. https://doi.org/10.1006/abio.1976.9999
Chen C (2016) Cost analysis of plant micropropagation of Phalaenopsis. Plant Cell Tissue Organ Cult 126:167–175. https://doi.org/10.1007/s11240-016-0987-4
Cheynier V, Comte G, Davies KM, Lattanzio V, Martens S (2013) Plant phenolics: Recent advances on their biosynthesis, genetics, and ecophysiology. Plant Physiol Biochem 72:1–20. https://doi.org/10.1016/j.plaphy.2013.05.009
Debergh PC (1983) Effects of agar brand and concentration on the tissue culture medium. Physiol Plant 59:270–276. https://doi.org/10.1111/j.1399-3054.1983.tb00770.x
Ferreira LT, de Araújo Silva MM, Ulisses C, Camara TR, Willadino L (2017) Using LED lighting in somatic embryogenesis and micropropagation of an elite sugarcane variety and its effect on redox metabolism during acclimatization. Plant Cell Tissue Organ Cult 128:211–221. https://doi.org/10.1007/s11240-016-1101-7
Ghaderi N, Hatami M, Aliakbar Mozafari A, Siosehmardeh A (2018) Change in antioxidant enzymes activity and some morpho-physiological characteristics of strawberry under long-term salt stress. Physiol Mol Biol Plants 24:833–843. https://doi.org/10.1007/s12298-018-0535-2
Ghorbani A (2005) Studies on pharmaceutical ethnobotany in the region of Turkmen Sahra, north of Iran: (Part 1): general results. J Ethnopharmacol 102:58–68. https://doi.org/10.1016/j.jep.2005.05.035
Grzegorczyk I, Wysokińska H (2008) Liquid shoot culture of Salvia officinalis L. for micropropagation and production of antioxidant compounds; effect of triacontanol. Acta Soc Bot Pol 77:99–104. https://doi.org/10.5586/asbp.2008.013
Grzegorczyk-Karolak I, Kiss AK (2018) Determination of the phenolic profile and antioxidant properties of Salvia viridis L. shoots: A comparison of aqueous and hydroethanolic extracts. Molecules 23:1468. https://doi.org/10.3390/molecules23061468
Grzegorczyk-Karolak I, Rytczak P, Bielecki S, Wysokińska H (2017) The influence of liquid systems for shoot multiplication, secondary metabolite production and plant regeneration of Scutellaria alpina. Plant Cell Tissue Organ Cult 128:479–486. https://doi.org/10.1007/s11240-016-1126-y
Grzegorczyk-Karolak I, Kuźma Ł, Lisiecki P, Kiss AK (2019) Accumulation of phenolic compounds in different in vitro cultures of Salvia viridis L. and their antioxidant and antimicrobial potential. Phytochem Lett 30:324–332. https://doi.org/10.1016/j.phytol.2019.02.016
Grzegorczyk-Karolak I, Hnatuszko-Konka K, Zarzycka M, Kuźma Ł (2020) The stimulatory effect of purine-type cytokinins on proliferation and polyphenolic compound accumulation in shoot culture of Salvia viridis. Biomolecules 10(2):178. https://doi.org/10.3390/biom10020178
Grzegorczyk-Karolak I, Grąbkowska R, Piątczak E (2021) Plant liquid cultures as a source of bioactive metabolites. In: Ekiert HM, Goyal S (eds) Ramwat KG. Plant Cell and Tissue Differentiation and Secondary Metabolites, Fundamentals and Applications. Springer, pp 743–771
Gupta SD, Sahoo TK (2015) Light emitting diode (LED) induced alteration of oxidative events during in vitro shoot organogenesis of Curculigo orchioides Gaertn. Acta Physiol Plant 37:233. https://doi.org/10.1007/s11738-015-1990-9
Hammock HA, Kopsell DA, Sams CE (2020) Supplementary blue and red LED narrowband wavelengths improve biomass yield and nutrient uptake in hydroponically grown basil. HortScience 55:1888–1897. https://doi.org/10.21273/HORTSCI15267-20
Hasnain A, Naqvi SAH, Ayesha SI, Khalid F, Ellahi M, Iqbal S, Hassan MZ, Abbas A, Adamski R, Markowska D, Baazeem A, Mustafa G, Moustafa M, Hasan ME, Abdelhamid M (2022) Plants in vitro propagation with its applications in food, pharmaceuticals and cosmetic industries; current scenario and future approaches. Front Plant Sci 13:1009395. https://doi.org/10.3389/fpls.2022.1009395
Ighodaro OM, Akinloye OA (2018) First line defence antioxidants-superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX): Their fundamental role in the entire antioxidant defence grid. Alexandria J Med 54:287–293. https://doi.org/10.1016/j.ajme.2017.09.001
Isah T, Umar S, Mujib A, Sharma MP, Rajasekharan PE, Zafar N, Frukh A (2018) Secondary metabolism of pharmaceuticals in the plant in vitro cultures: strategies, approaches, and limitations to achieving higher yield. Plant Cell Tissue Organ Cult 132:239–265. https://doi.org/10.1007/s11240-017-1332-2
Ivanova M, van Staden J (2011) Influence of gelling agent and cytokinins on the control of hyperhydricity in Aloe polyphylla. Plant Cell Tissue Organ Cult 104:13–21. https://doi.org/10.1007/s11240-010-9794-5
Jeong BR, Sivanesan I (2015) Direct adventitious shoot regeneration, in vitro flowering, fruiting, secondary metabolite content and antioxidant activity of Scrophularia takesimensis Nakai. Plant Cell Tissue Organ Cult 123:607–618. https://doi.org/10.1007/s11240-015-0864-6
Kevers C, Franck T, Strasser RJ, Dommes J, Gaspar T (2004) Hyperhydricity of micropropagated shoots: a typically stress-induced change of physiological state. Plant Cell Tissue Organ Cult 77:181–191. https://doi.org/10.1023/B:TICU.0000016825.18930.e4
Krzemińska M, Hnatuszko-Konka K, Weremczuk-Jeżyna I, Owczarek-Januszkiewicz A, Ejsmont W, Olszewska MA, Grzegorczyk-Karolak I (2023) Effect of light conditions on polyphenol production in transformed shoot culture of Salvia bulleyana Diels. Molecules 28:4603. https://doi.org/10.3390/molecules28124603
Lima CC, Cajueiro Gurgel ES, Borges EEL (2021) Antioxidant enzyme activity in germination of Dalbergia spruceana seeds under different temperatures. J Seed Sci 43:e202143006. https://doi.org/10.1590/2317-1545v43244385
Lobiuc V, Vasilache O, Pintilie T, Stolen M, Burduera M, Oroian MM, Zamirache MM (2017) Blue and red illumination improves growth and bioactive compounds contents in acyanic and cyanic Ocimmum basilicum L. microgreens. Molecules 22:2111. https://doi.org/10.3390/molecules22122111
Makowski W, Królicka A, Tokarz B, Szopa A, Ekiert H, Tokarz KM (2023) Temporary immersion bioreactors as a useful tool for obtaining high productivity of phenolic compounds with strong antioxidant properties from Pontechium maculatum. Plant Cell Tissue Organ Cult 153(3):525–537. https://doi.org/10.1007/s11240-023-02487-6
Manivannan A, Soundararajan P, Halimah N, Ko CH, Jeong BR (2015) Blue LED light enhances growth, phytochemical contents, and antioxidant enzyme activities of Rehmannia glutinosa cultured in vitro. Hortic Environ Biotechnol 56:105–113. https://doi.org/10.1007/s13580-015-0114-1
Mengxi L, Zhigang X, Yang Y, Yijie F (2011) Effects of different spectral lights on Oncidium PLBs induction, proliferation, and plant regeneration. Plant Cell Tissue Organ Cult 106:1–10. https://doi.org/10.1007/s11240-010-9887-1
Miler N, Kulus D, Woźny A, Rymarz D, Hajzer M, Wierzbowski K, Nelke R, Szeffs L (2019) Application of wide-spectrum light-emitting diodes in micropropagation of popular ornamental plant species: A study on plant quality and cost reduction. In Vitro Cell Dev Biol Plant 55:99–108. https://doi.org/10.1007/s11627-018-9939-5
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
Nhut DT, Huy NP, Tai NT, Nam NB, Luan VQ, Tung HVT, Vinh HT, BT, Luan TC, (2015) Light-emitting diodes and their potential in callus growth, plantlet development and saponin accumulation during somatic embryogenesis of Panax vietnamensis Ha et Grushv. Biotechnol Biotechnol Equip 29:299–308. https://doi.org/10.1080/13102818.2014.1000210
Ouzounis T, Razi Parjikolaei B, Fretté X, Rosenqvist E, Ottosen CO (2015) Predawn and high intensity application of supplemental blue light decreases the quantum yield of PSII and enhances the amount of phenolic acids, flavonoids, and pigments in Lactuca sativa. Front Plant Sci 6:19. https://doi.org/10.3389/fpls.2015.00019
Park WT, Yeo SK, Sathasivam R, Park JS, Kim JK, Park SU (2020) Influence of light-emitting diodes on phenylpropanoid biosynthetic gene expression and phenylpropanoid accumulation in Agastache rugosa. Appl Biol Chem 63:1–9. https://doi.org/10.1186/s13765-020-00510-4
Pati PK, Kaur J, Singh P (2011) A liquid culture system for shoot proliferation and analysis of pharmaceutically active constituents of Catharanthus roseus (L.) G. Don Plant Cell Tissue Organ Cult 105:299–307. https://doi.org/10.1007/s11240-010-9868-4
Pedroso RCN, Branquinho NAA, Hara AC, Costa AC, Silva FG, Pimenta LP, Silva MLA, Cunha WR, Paulleti PM, Januario AH (2017) Impact of light quality on flavonoid production and growth of Hyptis marrubioides seedlings cultivated in vitro. Rev Bras Farmacogn 27:466–470. https://doi.org/10.1016/j.bjp.2016.12.004
Piątczak E, Wielanek M, Wysokińska H (2005) Liquid culture system for shoot multiplication and secoiridoid production in micropropagated plants of Centaurium erythraea Rafn. Plant Sci 168:431–437. https://doi.org/10.1016/j.plantsci.2004.08.013
Prasad VSS, Gupta SD (2006) In vitro shoot regeneration of gladiolus in semi-solid agar versus liquid cultures with support systems. Plant Cell Tissue Organ Cult 87:263–271. https://doi.org/10.1007/s11240-006-9160-9
Rungsimakan S, Rowan MG (2014) Terpenoids, flavonoids and caffeic acid derivatives from Salvia viridis L. cvar. Blue Jeans Phytochemistry 108:177–188. https://doi.org/10.1016/j.phytochem.2014.08.029
Sales E, Nebauer SG, Arrilaga I, Segura J (2002) Plant hormones and Agrobacterium tumefaciens strain 82139 induce efficient plant regeneration in the cardenolide - producing plant Digitalis minor. J Plant Physiol 159:9–16. https://doi.org/10.1078/0176-1617-00534
Sandal I, Bhattacharya A, Ahuja PS (2001) An efficient liquid culture system for tea proliferation. Plant Cell Tissue Organ Cult 65:75–80. https://doi.org/10.1023/A:1010662306067
Savio LEB, Astarita LV, Santarém ER (2012) Secondary metabolism in micropropagated Hypericum perforatum L. grown in non-aerated liquid medium. Plant Cell Tissue Organ Cult 108:465–472. https://doi.org/10.1007/s11240-011-0058-9
Sharma A, Shahzad B, Rehman A, Bhardwaj R, Landi M, Zheng B (2019) Response of phenylpropanoid pathway and the role of polyphenols in plants under abiotic stress. Molecules 24:2452. https://doi.org/10.3390/molecules24132452
Shohael AM, Chakrabarty D, Yu KW, Hahn EJ, Paek KY (2005) Application of bioreactor system for large-scale production of Eleutherococcus sessiliflorus somatic embryos in an air-lift bioreactor and production of eleutherosides. J Biotechnol 120:228–236. https://doi.org/10.1016/j.jbiotec.2005.06.010
Sirivarasai J, Kaojarern S, Chanprasertyothin S, Panpunuan P, Petchpoung K, Tatsaneeyapant A, Yoovathaworn K, Sura T, Kaojarern S, Sritara P (2015) Environmental lead exposure, catalase gene, and markers of antioxidant and oxidative stress relation to hypertension: An analysis based on the EGAT study. Biomed Res Int 2015:856319. https://doi.org/10.1155/2015/856319
Szopa A, Kokotkiewicz A, Marzec-Wróblewska U, Bucinski A, Luczkiewicz M, Ekiert H (2016) Accumulation of dibenzocyclooctadiene lignans in agar cultures and in stationary and agitated liquid cultures of Schisandra chinensis (Turcz.) Baill. Appl Microbiol Biotechnol 100:3965–3977. https://doi.org/10.1007/s00253-015-7230-9
Thaniarasu R, Kumar TS, Rao MV (2015) In vitro propagation of Plectranthus bourneae gamble? An endemic red listed plant. Plant Tissue Cult Biotechnol 25:273–284
Tutin TG, Heywood VH, Burges NA, Moore DM, Valentine DH, Walters SM, Webb DA (eds) (1972) Flora europaea 3. diapensiacea to myoporaceae. Cambridge University Press, Cambridge, pp 192.
Ullah MA, Tungmunnithum D, Garros L, Hano C, Abbasi BH (2019) Monochromatic lights-induced trends in antioxidant and antidiabetic polyphenol accumulation in in vitro callus cultures of Lepidium sativum L. J Photochem Photobiol B Biol 196:111505. https://doi.org/10.1016/j.jphotobiol.2019.05.002
Vaidya BN, Asanakunov B, Shahin L, Jernigan HL, Joshee N, Dhekney SA (2019) Improving micropropagation of Mentha×piperita L. using a liquid culture system. In Vitro Cell Dev Biol Plant 55:71–80. https://doi.org/10.1007/s11627-018-09952-4
Weremczuk-Jeżyna I, Hnatuszko-Konka K, Lebelt L, Grzegorczyk-Karolak I (2021a) The protective function and modification of secondary metabolite accumulation in response to light stress in Dracocephalum forrestii shoots. Int J Mol Sci 22:7965. https://doi.org/10.3390/ijms22157965
Weremczuk-Jeżyna I, Kuźma Ł, Grzegorczyk-Karolak I (2021b) The effect of different light treatments on morphogenesis, phenolic compound accumulation and antioxidant potential of Dracocephalum forrestii transformed shoots cultured in vitro. J Photochem Photobiol B Biol 224:112329
Ye S, Shao Q, Xu M, Li S, Wu M, Tan X, Su L (2017) Effects of light quality on morphology, enzyme activities, and bioactive compound contents in Anoectochilus roxburghii. Front Plant Sci 8:857. https://doi.org/10.3389/fpls.2017.00857
Zhang S, Ma J, Zou H, Zhang L, Li S, Wang Y (2020) The combination of blue and red LED light improves growth and phenolic acid contents in Salvia miltiorrhiza Bunge. Ind Crop Prod 158:112959. https://doi.org/10.1016/j.indcrop.2020.112959
Funding
This work was supported by the Medical University of Lodz, grant No. 503/3–012-01/503–31-001.
Author information
Authors and Affiliations
Contributions
I.G.-K. designed the experiment. I.G.-K. obtained the biological material, performed experiments, collected, interpreted, and discussed the data. I.W.-J. performed the analysis of antioxidant enzyme activity. P.T. determined lighting conditions and contributed to data acquisition. I.G-K. performed the statistical analysis and prepared the graphical part of the manuscript. The first draft of the manuscript was written by I.G.-K. and all authors checked and corrected the manuscript. All authors read and approved the final manuscript.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Communicated by Sergio J. Ochatt.
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
Grzegorczyk-Karolak, I., Tabaka, P. & Weremczuk-Jeżyna, I. Enhancing polyphenol yield in Salvia viridis L. shoot culture through liquid medium optimization and light spectrum manipulation. Plant Cell Tiss Organ Cult 156, 88 (2024). https://doi.org/10.1007/s11240-024-02713-9
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s11240-024-02713-9