Abstract
Salvia abrotanoides (Kar.) Sytsma is a medicinal plant that contains phenolic acids, especially rosmarinic acid (RA), with a wide variety of health benefits. In this research, the stimulatory effects of sodium nitroprusside (SNP), a donor of nitric oxide (NO), were examined on phenolic acids accumulation in the shoot cultures of the species after different times of exposure. The developed shoots on single-nodal explants in Murashige and Skoog solid medium supplemented with 0.5 mg L−1 kinetin and 0.05 mg L−1 indole-3-acetic acid were treated with SNP at concentrations of 25, 50 and 100 µM. The highest content of RA (10.45 ± 0.40 mg g−1 DW) was measured 144 h after elicitation of the shoots with 100 µM SNP. The maximum values of salvianolic acid A (Sal-A) (0.065 ± 0.00 mg g−1 DW) and salvianolic acid B (Sal-B) (0.42 ± 0.01 mg g−1 DW) were obtained in the shoots after 96 and 144 h exposure to 50 and 25 μM SNP, respectively. Also, elicitation with SNP at different concentrations significantly upregulated the crucial genes (PAL, TAT, RAS and CYP98A14) involved in phenolic acids biosynthesis in the shoots with distinct patterns, although no strong correlations were observed between transcription levels of the genes and phenolic acids accumulation. The findings of this study provide beneficial information about the impact of NO as an effective elicitor, which could be valuable for the in vitro improvement of phenolic acids production in S. abrotanoides.
Key Message
The results demonstrated that the elicitation with nitric oxide upregulated critical genes in the biosynthetic pathways and led to phenolic acids accumulation in the shoots of Salvia abrotanoides (Kar.) Sytsma.
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Abbreviations
- KIN:
-
Kinetin
- IAA:
-
Indole-3-acetic acid
- FSI:
-
Frequency of shoot initiation
- MSN:
-
Mean shoot number
- MSL:
-
Mean shoot length
- NO:
-
Nitric oxide
- SNP:
-
Sodium nitroprusside
- TPC:
-
Total phenolic content
- TFC:
-
Total flavonoid content
- RA:
-
Rosmarinic acid
- Sal-A:
-
Salvianolic acid A
- Sal-B:
-
Salvianolic acid B
- PAL:
-
Phenylalanine ammonia-lyase
- TAT:
-
Tyrosine aminotransferase
- RAS:
-
Rosmarinic acid synthase
- CYP98A14:
-
Cytochrome P450-dependent monooxygenase
References
Alizadeh Z, Farimani MM, D’Ambola M, Marzocco S, Rapa SF, Braca A, De Tommasi N (2019) New abietane-type diterpenes from Perovskia abrotanoides and their anti-inflammatory activity. Planta Med 85:355. https://doi.org/10.1055/s-0039-3400067
Ashraf SN, Zubair M, Rizwan K, Tareen RB, Rasool N, Zia-Ul-Haq M, Ercisli S (2014) Compositional studies and Biological activities of Perovskia abrotanoides Kar. oils. Biol Res 47:1–9. https://doi.org/10.1186/0717-6287-47-12
Bassolino L, Giacomelli E, Giovanelli S, Pistelli L, Cassetti A, Damonte G, Bisio A, Ruffoni B (2015) Tissue culture and aromatic profile in Salvia dolomitica Codd. Plant Cell Tissue Org (PCTOC) 121:83–95. https://doi.org/10.1007/s11240-014-0681-3
Beikmohammadi M (2012) The evaluation of medicinal properties of Perovskia abrotanoides Karel. Middle-East J Sci Res 11:189–193
Bellin D, Asai S, Delledonne M, Yoshioka H (2013) Nitric oxide as a mediator for defense responses. MPMI 26:271–277. https://doi.org/10.1094/MPMI-09-12-0214-CR
Chaturvedi H, Jain M, Kidwai N (2007) Cloning of medicinal plants through tissue culture: a review. IJEB 45:937–948
Di P, Zhang L, Chen J, Tan H, Xiao Y, Dong X, Zhou X, Chen W (2013) 13C tracer reveals phenolic acids biosynthesis in hairy root cultures of Salvia miltiorrhiza. ACS Chem Biol 8:1537–1548. https://doi.org/10.1021/cb3006962
Dong J, Wan G, Liang Z (2010) Accumulation of salicylic acid-induced phenolic compounds and raised activities of secondary metabolic and antioxidative enzymes in Salvia miltiorrhiza cell culture. J Biotechnol 148:99–104. https://doi.org/10.1016/j.jbiotec.2010.05.009
Dowom SA, Abrishamchi P, Radjabian T, Salami SA (2017) Enhanced phenolic acids production in regenerated shoot cultures of Salvia virgata Jacq. after elicitation with Ag+ ions, methyl jasmonate and yeast extract. Ind Crops Prod 103:81–88. https://doi.org/10.1016/j.indcrop.2017.03.043
Ebrahimi S, Zaker A, Abrishamchi P, Bahrami AR, Ganjeali A, Sodagar N (2017) Hairy root induction and secondary metabolite production in Perovskia abrotanoides Karel. J Plant Process Funct 6:17–26
Ejtahed RS, Radjabian T, Tafreshi SAH (2015) Expression analysis of phenylalanine ammonia lyase gene and rosmarinic acid production in Salvia officinalis and Salvia virgata shoots under salicylic acid elicitation. Appl Biochem Biotechnol 176:1846. https://doi.org/10.1007/s12010-015-1682-3
El-beltagi HS, Ahmed OK, Hegazy AE (2015) Molecular role of nitric oxide in secondary products production in Ginkgo biloba cell suspension culture. Not Bot Horti Agrobo 43:12–18. https://doi.org/10.15835/nbha4319660
Erişen S, Kurt-Gür G, Servi H (2020) In vitro propagation of Salvia sclarea L. by meta-Topolin, and assessment of genetic stability and secondary metabolite profiling of micropropagated plants. Ind Crops Prod 157:112892. https://doi.org/10.1016/j.indcrop.2020.112892
Farouk S, Al-Huqail AA (2020) Sodium nitroprusside application regulates antioxidant capacity, improves phytopharmaceutical production and essential oil yield of marjoram herb under drought. Ind Crops Prod 158:113034. https://doi.org/10.1016/j.indcrop.2020.113034
Fatemi F, Abdollahi MR, Mirzaie-asl A, Dastan D, Garagounis C, Papadopoulou K (2019) Identification and expression profiling of rosmarinic acid biosynthetic genes from Satureja khuzistanica under carbon nanotubes and methyl jasmonate elicitation. Plant Cell Tissue Org (PCTOC) 136:561–573. https://doi.org/10.1007/s11240-018-01537-8
Fu R, Shi M, Deng C, Zhang Y, Zhang X, Wang Y, Kai G (2020) Improved phenolic acid content and bioactivities of Salvia miltiorrhiza hairy roots by genetic manipulation of RAS and CYP98A14. Food Chem 331:127365. https://doi.org/10.1016/j.foodchem.2020.127365
Georgiev V, Pavlov A (2017) Salvia biotechnology. Springer, Switzerland
Ghaderi S, Ebrahimi SN, Ahadi H, Moghadam SE, Mirjalili MH (2019) In vitro propagation and phytochemical assessment of Perovskia abrotanoides Karel. (Lamiaceae): a medicinally important source of phenolic compounds. Biocatal Agric Biotechnol 19:101113. https://doi.org/10.1016/j.bcab.2019.101113
Ghanbar T, Hosseini B, Jabbarzadeh Z, Farokhzad A, Sharafi A (2016) High-frequency in vitro direct shoots regeneration from axillary nodal and shoot tip explants of clary sage (Salvia sclarea L.). Bulg J Agric Sci 22:73–78
Gorelick J, Bernstein N (2014) Elicitation: an underutilized tool in the development of medicinal plants as a source of therapeutic secondary metabolites. Adv Agron 124:201–230. https://doi.org/10.1016/B978-0-12-800138-7.00005-X
Grzegorczyk I, Wysokińska H (2009) The effect of methyl jasmonate on production of antioxidant compounds in shoot cultures of Salvia officinalis L. Herba Pol 55:238–243
Grzegorczyk I, Kuźma Ł, Lisiecki P, Kiss A (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
Guo H, Dang X, Dong J (2014) Hydrogen peroxide and nitric oxide are involved in salicylic acid-induced salvianolic acid B production in Salvia miltiorrhiza cell cultures. Molecules 19:5913–5924. https://doi.org/10.3390/molecules19055913
Habtemariam S (2018) Molecular pharmacology of rosmarinic and salvianolic acids: potential seeds for Alzheimer’s and vascular dementia drugs. Int J Mol Sci 19:458–482. https://doi.org/10.3390/ijms19020458
Hao G, Du X, Zhao F, Shi R, Wang J (2009) Role of nitric oxide in UV-B-induced activation of PAL and stimulation of flavonoid biosynthesis in Ginkgo biloba callus. Plant Cell Tissue Org (PCTOC) 97:175–185. https://doi.org/10.1007/s11240-009-9513-2
Hou X, Shao F, Ma Y, Lu SH (2013) The phenylalanine ammonia-lyase gene family in Salvia miltiorrhiza: genome-wide characterization, molecular cloning and expression analysis. Mol Biol Rep 40:4301–4310. https://doi.org/10.1007/s11033-013-2517-3
Isah T, Umar S, Mujib A, Sharma MP, Rajasekharan P, 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 Tiss Org (PCTOC) 132:239–265. https://doi.org/10.1007/s11240-017-1332-2
Jamzad Z (2012) Flora of Iran: Lamaiceae. Research Institute of Forests and Rangelands, Tehran
Jan T, Naqvi B, Hazrat A, Qadri R, Nisar M, Khan N, Nawaz MA, Pervez S (2020) Impact of hormones on the proliferation of shoots and initiation of roots in Salvia santolinifolia (Boiss), a high value medicinal herb. Pak J Sci Ind Res B: Biol Sci 63:30–36. https://doi.org/10.52763/PJSIR.BIOL.SCI.63.1.2020.30.36
Jawahar G, Punita D, Rajasheker G, Manoharachary C, Venkatachalam P, Kishor PK (2018) Feeding elicitors and precursors enhance colchicine accumulation in morphogenic cultures of Gloriosa superba L. Plant Cell Tissue Org (PCTOC) 135:235–245. https://doi.org/10.1007/s11240-018-1459-9
Jiao C, Yang R, Zhou Y, Gu Z (2016) Nitric oxide mediates isoflavone accumulation and the antioxidant system enhancement in soybean sprouts. Food Chem 204:373–380. https://doi.org/10.1016/j.foodchem.2016.02.147
Khezerluo M, Hosseini B, Amiri J (2018) Sodium nitroprusside stimulated production of tropane alkaloids and antioxidant enzymes activity in hairy root culture of Hyoscyamus reticulatus L. Acta Biol Hung 69:437–448. https://doi.org/10.1556/018.69.2018.4.6
Kong JQ (2015) Phenylalanine ammonia-lyase, a key component used for phenylpropanoids production by metabolic engineering. RSC Adv 5:62587–62603. https://doi.org/10.1039/C5RA08196C
Kumar N, Goel N (2019) Phenolic acids: Natural versatile molecules with promising therapeutic applications. Biotechnol Rep 24:1–10. https://doi.org/10.1016/j.btre.2019.e00370
Li G, Zhu S, Wu W, Zhang C, Peng Y, Wang Q, Shi J (2017a) Exogenous nitric oxide induces disease resistance against Monilinia fructicola through activating the phenylpropanoid pathway in peach fruit. J Sci Food Agric 97:3030–3038. https://doi.org/10.1002/jsfa.8146
Li X, Zhang L, Ahammed GJ, Li ZX, Wei JP, Shen C, Yan P, Zhang LP, Han WY (2017b) Nitric oxide mediates brassinosteroid-induced flavonoid biosynthesis in Camellia sinensis L. J Plant Physiol 214:145–151. https://doi.org/10.1016/j.jplph.2017.04.005
Li Q, Feng J, Chen L, Xu Z, Zhu Y, Wang Y, Xiao Y, Chen J, Zhou Y, Tan H (2019) Genome-wide identification and characterization of Salvia miltiorrhiza laccases reveal potential targets for salvianolic acid B biosynthesis. Front Plant Sci 10:1–15. https://doi.org/10.3389/fpls.2019.00435
Lubbe A, Verpoorte R (2011) Cultivation of medicinal and aromatic plants for specialty industrial materials. Ind Crops Prod 34:785–801. https://doi.org/10.1016/j.indcrop.2011.01.019
Mahendran G, Iqbal Z, Kumar D, Verma SK, Rout PK, ur Rahman L, (2021) Enhanced gymnemic acids production in cell suspension cultures of Gymnema sylvestre (Retz.) R. Br. ex Sm. through elicitation. Ind Crops Prod 162:113234. https://doi.org/10.1016/j.indcrop.2020.113234
Marchev A, Haas C, Schulz S, Georgiev V, Steingroewer J, Bley T, Pavlov A (2014) Sage in vitro cultures: a promising tool for the production of bioactive terpenes and phenolic substances. Biotechnol Lett 36:211–221. https://doi.org/10.1007/s10529-013-1350-z
Misic D, Grubisic D, Konjevic R (2006) Micropropagation of Salvia brachyodon through nodal explants. Biol Plant 50:473–476. https://doi.org/10.1007/s10535-006-0074-5
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
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
Nourozi E, Hosseini B, Maleki R, Mandoulakani BA (2019) Pharmaceutical important phenolic compounds overproduction and gene expression analysis in Dracocephalum kotschyi hairy roots elicited by SiO2 nanoparticles. Ind Crops Prod 133:435–446. https://doi.org/10.1016/j.indcrop.2019.03.053
Pesaraklu A, Radjabian T, Salami SA (2021) Methyl jasmonate and Ag+ as effective elicitors for enhancement of phenolic acids contents in Salvia officinalis and Salvia verticillata, as two traditional medicinal plants. S Afr J Bot 141:105–115. https://doi.org/10.1016/j.sajb.2021.04.032
Petersen M (2013) Rosmarinic acid: new aspects. Phytochem Rev 12:207–227. https://doi.org/10.1007/s11101-013-9282-8
Petrova M, Nikolova M, Dimitrova L, Zayova E (2015) Micropropagation and evaluation of flavonoid content and antioxidant activity of Salvia officinalis L. Genet Plant Physiol 5:48–60
Rahmani N, Radjabian T, Soltani BM (2020) Impacts of foliar exposure to multi-walled carbon nanotubes on physiological and molecular traits of Salvia verticillata L., as a medicinal plant. Plant Physiol Biochem 150:27–38. https://doi.org/10.1016/j.plaphy.2020.02.022
Ramirez-Estrada K, Vidal-Limon H, Hidalgo D, Moyano E, Golenioswki M, Cusidó RM, Palazon J (2016) Elicitation, an effective strategy for the biotechnological production of bioactive high-added value compounds in plant cell factories. Molecules 21:182. https://doi.org/10.3390/molecules21020182
Rani V, Raina S (2000) Genetic fidelity of organized meristem-derived micropropagated plants: a critical reappraisal. In Vitro Cell Dev Biol Plant 36:319–330. https://doi.org/10.1007/s11627-000-0059-6
Rezaei A (2018) Nitric oxide increased the rosmarinic acid and essential oil production in in vitro-cultured Melissa officinalis. J Med Plants 65:61–72
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
Shi M, Huang F, Deng Ch, Wang Y, Kia G (2018) Bioactivities, biosynthesis and biotechnological production of phenolic acids in Salvia miltiorrhiza. Crit Rev Food Sci Nutr 56:953–964. https://doi.org/10.1080/10408398.2018.1474170
Siddiqui MH, Al-Whaibi MH, Basalah MO (2011) Role of nitric oxide in tolerance of plants to abiotic stress. Protoplasma 248:447–455. https://doi.org/10.1007/s00709-010-0206-9
Singleton VL, Orthofer R, Lamuela-Raventós RM (1999) Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent. Meth Enzymol 299:152–178. https://doi.org/10.1016/S0076-6879(99)99017-1
Skała E, Wysokińska H (2004) In vitro regeneration of Salvia nemorosa L. from shoot tips and leaf explants. In Vitro Cell Dev Biol Plant 40:596–602. https://doi.org/10.1079/IVP2004580
Skała E, Kalemba D, Wajs A, Róźalski M, Krajewska U, Rózalska B, Wieckowska-Szakiel M, Wysokinska H (2007) In vitro propagation and chemical and biological studies of the essential oil of Salvia przewalskii Maxim. Zeitschrift Für Naturforschung C 62:839–848. https://doi.org/10.1515/znc-2007-11-1212
Song Z, Li X (2015) Expression profiles of rosmarinic acid biosynthesis genes in two Salvia miltiorrhiza lines with differing water-soluble phenolic contents. Ind Crops Prod 71:24–30. https://doi.org/10.1016/j.indcrop.2015.03.081
Song J, Wang Z (2011) RNAi-mediated suppression of the phenylalanine ammonia-lyase gene in Salvia miltiorrhiza causes abnormal phenotypes and a reduction in rosmarinic acid biosynthesis. J Plant Res 124:183–192. https://doi.org/10.1007/s10265-010-0350-5
Stafiniak M, Slusarczyk S, Pencakowski B, Matkowski A, Rahimmalek M, Bielecka M (2021) Seasonal variations of rosmarinic acid and its glucoside and expression of genes related to their biosynthesis in two medicinal and aromatic species of Salvia subg. Perovskia. Biology 10:458–476. https://doi.org/10.3390/biology10060458
Tossi V, Amenta M, Lamattina L, Cassia R (2011) Nitric oxide enhances plant ultraviolet-B protection up-regulating gene expression of the phenylpropanoid biosynthetic pathway. Plant Cell Environ 34:909–921. https://doi.org/10.1111/j.1365-3040.2011.02289.x
Trivellini A, Lucchesini M, Maggini R, Mosadegh H, Villamarin TSS, Vernieri P, Mensuali-Sodi A, Pardossi A (2016) Lamiaceae phenols as multifaceted compounds: bioactivity, industrial prospects and role of “positive-stress.” Ind Crops Prod 83:241–254. https://doi.org/10.1016/j.indcrop.2015.12.039
Verma N, Shukla S (2015) Impact of various factors responsible for fluctuation in plant secondary metabolites. JARMAP 2:105–113. https://doi.org/10.1016/j.jarmap.2015.09.002
Wang JW, Wu JY (2005) Nitric oxide is involved in methyl jasmonate-induced defense responses and secondary metabolism activities of Taxus cells. Plant Cell Physiol 46:923–930. https://doi.org/10.1093/pcp/pci098
Wang B, Sun W, Li Q, Li Y, Luo H, Song J, Sun Ch, Qian J, Zhu Y, Hayward A, Xu H, Chen Sh (2015) Genome-wide identification of phenolic acid biosynthetic genes in Salvia miltiorrhiza. Planta 241:711–725. https://doi.org/10.1007/s00425-014-2212-1
Wang J, Xu J, Gong X, Yang M, Zhang C, Li M (2019) Biosynthesis, chemistry, and pharmacology of polyphenols from Chinese Salvia species: a review. Molecules 24:155. https://doi.org/10.3390/molecules24010155
Wu CH, Tewari RK, Hahn EJ, Paek KY (2007) Nitric oxide elicitation induces the accumulation of secondary metabolites and antioxidant defense in adventitious roots of Echinacea purpurea. J Plant Biol 50:636–643. https://doi.org/10.1007/BF03030607
Wu SJ, Qi JL, Zhang WJ, Liu SH, Xiao FH, Zhang MS, Xu GH, Zhao WG, Shi MW, Pang YJ (2009) Nitric oxide regulates shikonin formation in suspension-cultured Onosma paniculatum cells. Plant Cell Physiol 50:118–128. https://doi.org/10.1093/pcp/pcn178
Wu Q, Su N, Zhang X, Liu Y, Cui J, Liang Y (2016) Hydrogen peroxide, nitric oxide and UV RESISTANCE LOCUS8 interact to mediate UV-B-induced anthocyanin biosynthesis in radish sprouts. Sci Rep 6:1–12. https://doi.org/10.1038/srep29164
Xu M, Dong J (2005) Nitric oxide stimulates indole alkaloid production in Catharanthus roseus cell suspension cultures through a protein kinase-dependent signal pathway. Enzyme Microb Technol 37:49–53. https://doi.org/10.1016/j.enzmictec.2005.01.036
Yildiz M (2012) The prerequisite of the success in plant tissue culture: high frequency shoot regeneration. In: Leva A, Rinaldi MR (eds) Recent advances in plant in vitro culture. Intech, Rijeka, pp 63–90
Zaker A, Sykora C, Gössnitzer F, Abrishamchi P, Asili J, Mousavi SH, Wawrosch C (2015) Effects of some elicitors on tanshinone production in adventitious root cultures of Perovskia abrotanoides Karel. Ind Crops Prod 67:97–102. https://doi.org/10.1016/j.indcrop.2015.01.015
Zhang B, Zheng LP, Wang JW (2012) Nitric oxide elicitation for secondary metabolite production in cultured plant cells. Appl Microbiol Biotechnol 93:455–466. https://doi.org/10.1007/s00253-011-3658-8
Zhang JJ, Li XQ, Sun JW, Jin SH (2014a) Nitric oxide functions as a signal in ultraviolet-B-induced baicalin accumulation in Scutellaria baicalensis suspension cultures. Int J Mol Sci 15:4733–4746. https://doi.org/10.3390/ijms15034733
Zhang S, Yan Y, Wang B, Liang Z, Liu Y, Liu F, Qi Z (2014b) Selective responses of enzymes in the two parallel pathways of rosmarinic acid biosynthetic pathway to elicitors in Salvia miltiorrhiza hairy root cultures. J Biosci Bioeng 117:645–651. https://doi.org/10.1016/j.jbiosc.2013.10.013
Zhishen J, Mengcheng T, Jianming W (1999) The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals. Food Chem 64:555–559. https://doi.org/10.1016/S0308-8146(98)00102-2
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This work was supported by a research grant funded by Shahed University of Tehran for the plant physiology PhD thesis.
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Contributor 1 (FR) and Contributor 2 (TR) conceived this research and designed experiments; Contributor 1 (FR) performed experiments and analyses; Contributor 1 and Contributor 2 wrote the paper and Contributor 3 (PA) participated in the revisions of it. All authors read and approved the final manuscript.
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Rostami, F., Radjabian, T. & Abrishamchi, P. Enhancement of phenolic acids accumulation in Salvia abrotanoides (Kar.) Sytsma shoot cultures under elicitation with nitric oxide. Plant Cell Tiss Organ Cult 149, 441–453 (2022). https://doi.org/10.1007/s11240-022-02252-1
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DOI: https://doi.org/10.1007/s11240-022-02252-1