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. 2024 Jun 3;29(11):2626.
doi: 10.3390/molecules29112626.

Improvement of Bioactive Polyphenol Accumulation in Callus of Salvia atropatana Bunge

Izabela Grzegorczyk-Karolak  1 , Wiktoria Ejsmont  1 , Anna Karolina Kiss  2 , Przemyslaw Tabaka  3 , Wiktoria Starbała  1 , Marta Krzemińska  1
Affiliations

Improvement of Bioactive Polyphenol Accumulation in Callus of Salvia atropatana Bunge

Izabela Grzegorczyk-Karolak et al. Molecules. .

Abstract

Callus cultures of the Iranian medicinal plant Salvia atropatana were initiated from three-week-old seedlings on Murashige and Skoog (MS) medium supplemented with α-naphthaleneacetic acid (NAA) and various cytokinins. Although all tested hormonal variants of the medium and explant enabled callus induction, the most promising growth was noted for N-(2-chloro-4-pyridyl)-N'-phenylurea (CPPU)-induced calli. Three lines obtained on this medium (cotyledon line-CL, hypocotyl line-HL, and root line-RL) were preselected for further studies. Phenolic compounds in the callus tissues were identified using UPLC-MS (ultra-performance liquid chromatography-mass spectrometry) and quantified with HPLC (high-performance liquid chromatography). All lines exhibited intensive growth and contained twelve phenolic acid derivatives, with rosmarinic acid predominating. The cotyledon-derived callus line displayed the highest growth index values and polyphenol content; this was exposed to different light-emitting diodes (LED) for improving biomass accumulation and secondary metabolite yield. Under LED treatments, all callus lines exhibited enhanced RA and total phenolic content compared to fluorescent light, with the highest levels observed for white (48.5-50.2 mg/g dry weight) and blue (51.4-53.9 mg/g dry weight) LEDs. The selected callus demonstrated strong antioxidant potential in vitro based on the 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH), and ferric reducing antioxidant power (FRAP) tests. Our findings confirm that the S. atropatana callus system is suitable for enhanced rosmarinic acid production; the selected optimized culture provide high-quality plant-derived products.

Keywords: LEDs; abiotic stress; callus culture; light spectrum; line selection; phenolic acids; rosmarinic acid.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
Callus tissue on MS medium supplemented with 0.2 mg/L NAA and 2 mg/L CPPU; the initial subculture (subculture 0): callus from cotyledon (A), hypocotyl (B), and root (C) fragments; subculture 20: cotyledon- (D), hypocotyl- (E), and root- (F) derived line. The duration of the subculture was four weeks. Bar = 1 cm.
Figure 2
Figure 2
Growth of callus S. atropatana: RL (root line), CL (cotyledon line), and HL (hypocotyl) expressed as growth indices (GIs) of FW (fresh weight) and DW (dry weight). The values represent the mean ± standard error of three independent experiments. Means marked with the same letter for the same parameter were not significantly different (p < 0.05).
Figure 3
Figure 3
UPLC chromatogram of the extract of S. atropatana callus. The peak numbers are indicated in Table 2.
Figure 4
Figure 4
Total phenolic content in RL (root line), CL (cotyledon line), and HL (hypocotyl) callus cultivated on MS medium with 0.2 mg/L NAA and 2 mg/L CPPU after four weeks (subculture 18–20). The values represent the mean ± standard error of three independent experiments. Means marked with the same letter for the same parameter were not significantly different (p < 0.05).
Figure 5
Figure 5
Callus tissue of S. atropatana cultivated on MS medium supplemented with 0.2 mg/L NAA and 2 mg/L CPPU under W (white) (A), R (red) (B), B (blue) (C), R/B (70% red and 30% blue) (D) LEDs, FL (fluorescent lamps) (E), and in D (dark) (F) (subculture 33). The duration of the subculture was four weeks. Bar = 1 cm.
Figure 6
Figure 6
Growth of S. atropatana CL (cotyledon line) callus under different light conditions expressed as growth indices (GIs) of FW (fresh weight) and DW (dry weight). The duration of the subculture was four weeks. The values represent the mean ± standard error of three independent experiments. Means marked with the same letter for the same parameter were not significantly different (p < 0.05).
Figure 7
Figure 7
Polyphenol content in CL (cotyledon line) callus of S. atropatana cultivated on MS medium supplemented with 0.2 mg/L NAA and 2 mg/L CPPU under W (white), R (red), B (blue), R/B (70% red and 30% blue) LEDs, FL (fluorescent lamps), and in D (dark) after four weeks. CA—caffeic acid; CAH I, CAH II, CAH III—caffeic acid hexoside I, II, III; RAH I, RAH II—rosmarinic acid hexoside I, II; MRA—methyl rosmarinate; PRO I, PRO II—prolithospermic acid I, II; SAF I and II—salvianolic acid F isomers I and II; RA—rosmarinic acid; TPC—total polyphenol content. The values represent the mean ± standard error of three independent experiments. Means marked with the same letter for the same parameter were not significantly different (p < 0.05).
Figure 8
Figure 8
Relative spectral characteristics of the light emitted by the tested LEDs and fluorescent lamps.
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References

    1. Ramírez-Estrada K., Vidal-Limón H., Hidalgo D., Moyano E., Goleniowski M., Cusidó R.M., Palazon J. Elicitation, an effective strategy for the biotechnological production of bioactive high-added value compounds in plant cell factories. Molecules. 2016;21:182. doi: 10.3390/molecules21020182. - DOI - PMC - PubMed
    1. Espinosa-Leal C.A., Puente-Garza C.A., García-Lara S. In vitro plant tissue culture: Means for production of biological active compounds. Planta. 2018;248:1–18. doi: 10.1007/s00425-018-2910-1. - DOI - PMC - PubMed
    1. Murthy H.N., Lee E.J., Paek K.Y. Production of secondary metabolites from cell and organ cultures: Strategies and approaches for biomass improvement and metabolite accumulation. Plant Cell Tissue Organ Cult. 2014;118:1–16. doi: 10.1007/s11240-014-0467-7. - DOI
    1. Khan T., Ullah M.A., Garros L., Hano C., Abbasi B.H. Synergistic effects of melatonin and distinct spectral lights for enhanced production of anti-cancerous compounds in callus cultures of Fagonia indica. J. Photochem. Photobiol. B Biol. 2019;190:163–171. doi: 10.1016/j.jphotobiol.2018.10.010. - DOI - PubMed
    1. Ullah M.A., Tungmunnithum D., Garros L., Hano C., Abbasi B.H. Monochromatic lights-induced trends in antioxidant and antidiabetic polyphenol accumulation in in vitro callus cultures of Lepidium sativum L. J. Photochem. Photobiol. B Biol. 2019;196:111505. doi: 10.1016/j.jphotobiol.2019.05.002. - DOI - PubMed

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Grants and funding

This work was supported by the Medical University of Lodz, grant No. 503/3-012-01/503-31-001.

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