Antioxidant compounds in Salvia officinalis L. shoot and hairy root cultures in the nutrient sprinkle bioreactor

The study focused on the production of compounds with antioxidant activity in hairy root and shoot cultures of Salvia officinalis grown in laboratory-scale sprinkle nutrient bioreactors. HPLC analysis showed that production of rosmarinic acid in transformed roots (34.65 ±1.07 mg l-1) was higher that in shoot culture (26.24 ±0.48 mg l-1). In the latter diterpenoids: carnosic acid (1.74 ±0.02 mg l-1) and carnosol (1.34 ±0.01 mg l-1) were also found. Biomass accumulation after a growth period in the bioreactor was also studied. An 18-fold increase in hairy root biomass was recorded after 40 days of culture. In sage shoot culture, biomass increased 43 times after 21 days of bioreactor run. The current operating conditions of the bioreactor were not suitable for the propagation of Salvia officinalis mainly due to the hyperhydricity problem of leaves and stems.

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Liquid shoot culture of Salvia officinalis L. for micropropagation and production of antioxidant compounds; effect of triacontanol

Acta Societatis Botanicorum Poloniae ◽

10.5586/asbp.2008.013 ◽

2011 ◽

Vol 77 (2) ◽

pp. 99-104 ◽

Cited By ~ 15

Author(s):

Izabela Grzegorczyk ◽

Halina Wysokińska

Keyword(s):

Liquid Medium ◽

Rosmarinic Acid ◽

Shoot Culture ◽

Shoot Proliferation ◽

Shoot Multiplication ◽

Herbal Product ◽

Dry Weight ◽

Salvia Officinalis ◽

Carnosic Acid ◽

Antioxidant Compounds

Liquid shoot culture of Salvia officinalis L. in MS medium containing IAA (0.1 mg l-1) and BAP (0.45 mg l-1) was developed and evaluated in relation to shoot multiplication and antioxidant compound (carnosic acid, carnosol and rosmarinic acid) accumulation. In the liquid medium, on average, 3 new shoots per explant (shoot tip) were obtained within 3 weeks. The shoots produced 8.2±0.02 mg of diterpenoids and 31.2±0.29 mg of rosmarinic acid per gram of dry weight. Shoot proliferation and diterpenoid content increased when triacontanol (5, 10 or 20 pg l-1) was added to the liquid medium. In optimum conditions (at 20 pg l-1 TRIA) almost 7 shoots were formed per explant after 3 weeks. An increase in diterpenoid production (expressed as the sum of carnosol and carnosic acid) ranged from 30% to 50% and dependended on triacontanol concentration tested. The level of diterpenoids in triacontanol-treated shoots was similar to the content of compounds in commercial herbal product (dried leaves of S. officinalis) (10-12 mg g-1 dry wt). Triacontanol did not increase rosmarinic acid production, but the content of the phenolic as compound in shoots grown in liquid culture (31 mg g-1 dry wt) was even 24 times higher compared to samples of dried leaves of S. officinalis plants. We also demonstrated that the highest amounts of CA, Car and RA were accumulated in young, top parts of sage shoots. This observation could be useful for improving the selection of material for the extraction of natural antioxidants from S. officinalis.

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The effect of triacontanol on shoot multiplication and production of antioxidant compounds in shoot cultures of Salvia officinalis L.

Acta Societatis Botanicorum Poloniae ◽

10.5586/asbp.2006.002 ◽

2011 ◽

Vol 75 (1) ◽

pp. 11-15 ◽

Cited By ~ 6

Author(s):

Izabela Grzegorczyk ◽

Ireneusz Bilichowski ◽

Elżbieta Mikiciuk-Olasik ◽

Halina Wysokińska

Keyword(s):

Rosmarinic Acid ◽

Hplc Analysis ◽

Basal Medium ◽

Shoot Proliferation ◽

Shoot Multiplication ◽

Carnosic Acid ◽

Antioxidant Compounds ◽

Shoot Cultures ◽

Methanolic Extracts ◽

Salvia Officinalis L

This report describes the effect of triacontanol on shoot multiplication and production of antioxidant compounds (carnosic acid, carnosol and rosmarinic acid) in S. officinalis cultures grown on MS basal medium (agar solidified medium supplemented with 0.1 mg l-1 IAA, 0.45 mg l-1 BAP). It was found that shoot proliferation significantly increased when triacontanol at concentrations of 5, 10 or 20 µg l-1 was added to the medium. HPLC analysis of acetone and methanolic extracts of sage shoots showed that the production of diterpenoids, carnosic acid/carnosol ratio, as well as, contents of rosmarinic acid were also affected by the treatment with triacontanol. The highest stimulation effect of triacontanol was observed on the production of carnosol, where the treatment with 20 µg l l-1 increased the content of this diterpenoid 4.5-fold compared to that in the control (sage shoots growing on MS basal medium, only).

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In vitro cultures of Salvia officinalis L. as a source of antioxidant compounds

Acta Societatis Botanicorum Poloniae ◽

10.5586/asbp.2005.003 ◽

2011 ◽

Vol 74 (1) ◽

pp. 17-21 ◽

Cited By ~ 20

Author(s):

Izabela Grzegorczyk ◽

Ireneusz Bilichowski ◽

Elżbieta Mikiciuk-Olasik ◽

Halina Wysokińska

Keyword(s):

Cell Suspension ◽

Rosmarinic Acid ◽

Dry Weight ◽

Suspension Cultures ◽

In Vitro Cultures ◽

Salvia Officinalis ◽

Carnosic Acid ◽

Shoot Cultures ◽

Undifferentiated Callus

The concentrations of carnosic acid, carnosol and rosmarinic acid in different materials from differentiated (multiple shoot cultures and regenerated plants) and undifferentiated (callus and cell suspension) in vitro cultures of Salvia officinalis were determined by HPLC. The results suggested that diterpenoid (carnosic acid and carnosol) production is closely related to shoot differentiation. The highest diterpenoid yield (11.4 mg g-1 for carnosic acid and 1.1 mg g-1 for carnosol) was achieved in shoots of 10-week-old micropropagated plants. The levels were comparable to those found in shoots of naturally growing plants. Undifferentiated callus and cell suspension cultures produced only very low amounts of carnosol (ca. 0.05 mg g-1 of dry weight). In contrast, content of rosmarinic acid in callus and suspension cultures as well as shoots growing in vitro and in vivo was similar and ranged between 11.2 and 18.6 mg g-1 of dry weight.

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Extraction of Carnosic Acid and Carnosol from Sage (Salvia officinalis L.) Leaves by Supercritical Fluid Extraction and Their Antioxidant and Antibacterial Activity

Plants ◽

10.3390/plants8010016 ◽

2019 ◽

Vol 8 (1) ◽

pp. 16 ◽

Cited By ~ 7

Author(s):

Valentina Pavić ◽

Martina Jakovljević ◽

Maja Molnar ◽

Stela Jokić

Keyword(s):

Supercritical Fluid ◽

Flow Rate ◽

Acid Content ◽

Antioxidant Activities ◽

Salvia Officinalis ◽

Carnosic Acid ◽

Antioxidant Compounds ◽

Bacterial Strains ◽

Salvia Officinalis L ◽

Co2 Flow

Sage (Salvia officinalis L.) is a good source of antioxidant compounds, carnosic acid and carnosol being the prominent ones. Both are soluble in CO2, and our goal was to investigate the application of supercritical CO2 extraction to obtain sage extracts rich in these compounds. The effect of pressure, temperature, and CO2 flow rate on the carnosic acid and carnosol yield was studied. These variables were optimized by response surface methodology (RSM). The pressure significantly affected carnosol extraction, while the extraction of carnosic acid was affected by the pressure, temperature, and CO2 flow rate. Carnosic acid content varied from 0.29–120.0 µg mg−1, and carnosol content from 0.46–65.5 µg mg−1. The optimal conditions according to RSM were a pressure of 29.5 MPa, a temperature of 49.1°C, and a CO2 flow rate of 3 kg h−1, and the sage extract yield was calculated to be 6.54%, carnosic acid content 105 µg mg−1, and carnosol content 56.3 µg mg−1. The antioxidant activities of the sage extracts were evaluated by the scavenging activities of 2,2-diphenyl-1-picrylhydrazyl (DPPH). Sage extract obtained at 30 MPa and 40°C with 2 kg h−1 CO2 flow rate with a carnosic acid content of 72 µg mg−1 and carnosol content of 55 µg mg−1 exhibited the highest antioxidant activity (80.0 ± 0.68%) amongst the investigated supercritical fluid extracts at 25 µg mL−1 concentration. The antimicrobial properties of extracts were tested on four bacterial strains: Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, and Staphylococcus aureus. The extract with a carnosic acid content of 116 µg mg−1 and a carnosol content of 60.6 µg mg−1 was found to be the most potent agent against B. subtilis.

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High Yield of Bioactive Abietane Diterpenes in Salvia sclarea Hairy Roots by Overexpressing Cyanobacterial DXS or DXR Genes

Planta Medica ◽

10.1055/a-0895-5878 ◽

2019 ◽

Vol 85 (11/12) ◽

pp. 973-980 ◽

Cited By ~ 2

Author(s):

Mariacarmela Vaccaro ◽

Vivian Ocampo Bernal ◽

Nicola Malafronte ◽

Nunziatina De Tommasi ◽

Antonietta Leone

Keyword(s):

Hairy Root ◽

Hairy Roots ◽

Biological Activities ◽

Ectopic Expression ◽

Fold Increase ◽

Carnosic Acid ◽

High Yield ◽

Salvia Sclarea ◽

Cyanobacterial Genes ◽

Phosphate Synthase

AbstractAbietane diterpenoids, containing a quinone moiety, are synthesized in the roots of several Salvia species. Promising cytotoxicity and antiproliferative activities have been reported for these compounds in various cell and animal models. We have recently shown that aethiopinone, an o-naphto-quinone diterpene, produced in the roots of different Salvia species, is selectively cytotoxic against the A375 melanoma cell line. To enhance the synthesis of this abietane diterpenoid, we have engineered the plastidial 2-C-methyl-D-erythritol 4-phosphate-derived isoprenoid pathway in Salvia sclarea hairy roots by ectopic expression and plastid targeting of cyanobacterial genes encoding the 1-deoxy-D-xylulose 5-phosphate synthase or 1-deoxy-D-xylulose-5-phosphate reductoisomerase gene, the first two enzymatic steps of the plastidial MEP pathway, from which plant diterpenes primarily derive. Plastid-targeted expression of 1-deoxy-D-xylulose 5-phosphate synthase and 1-deoxy-D-xylulose-5-phosphate reductoisomerase proteins significantly enhanced the yield of aethiopinone by a 3-fold and about 6-fold increase, respectively. The accumulation of other abietane-type diterpenes (ferruginol, salvipisone, and carnosic acid), with interesting antiproliferative activity, was also increased. Compared to our previous data obtained by overexpressing the plant orthologous 1-deoxy-D-xylulose 5-phosphate synthase and 1-deoxy-D-xylulose-5-phosphate reductoisomerase genes in S. sclarea hairy roots, the results presented here confirm that the bacterial 1-deoxy-D-xylulose-5-phosphate reductoisomerase enzyme plays a major role than the DXS enzyme in the biosynthetic pathway of this class of compounds and that its ectopic expression does not conflict with active hairy root growth, resulting in a balanced trade-off between the transgenic hairy root final biomass and the increased content of o-naphto-quinone diterpenes, with interesting biological activities.

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The circulating floating bed reactor: effect of particle size distribution of the carrier on ammonia conversion

Water Science & Technology ◽

10.2166/wst.2000.0471 ◽

2000 ◽

Vol 41 (4-5) ◽

pp. 393-400 ◽

Cited By ~ 5

Author(s):

J.M. Garrido-Fernandez ◽

R. Méndez ◽

J.M. Lema ◽

V. Lazarova

Keyword(s):

Particle Size ◽

Size Distribution ◽

G Protein ◽

Biomass Concentration ◽

Biomass Accumulation ◽

Ammonia Removal ◽

Operating Conditions ◽

Nitrification Rate ◽

Ammonia Conversion ◽

Biomass Activity

Three Circulating Floating Bed Reactors (CFBR) R1, R2 and R3 with 20% v/v of a plastic carrier with different size distribution were operated to study the effect of the particles size of the carrier on biomass accumulation and nitrification performance. Operating conditions were similar in the three systems: ammonia concentrations around 50 mg-N–NH4+/ L, ammonia loading rates up to 1.2 kg N–NH4+/m3·d and temperatures between 14 and 27°C. Accumulation of nitrite was observed until day 65th. This w as result both of the inhibition of nitrite oxidation by free ammonia until day 20th and the insignificant accumulation of a biomass with low nitrite oxidising capacity between days 20 and 65th. Ammonia conversion rate and removal efficiency were higher in the reactor with lower particle size, R3 (nitrification rate of 1.1 kg N–NH4+/m3·d and ammonia removal of 97% at 16°C), than in R2 or R1 (nitrification rate of 1.0 kg N–NH4+/m3·d and ammonia removal of 90% at 16°C). The better efficiency in R3 was obtained as a result of the higher specific surface of the biofilm developed. Biomass activity was similar in the three reactors (2.2 and 1.12 g N/g protein · d at 30 and 15°C, respectively). Both the biomass evolution with time and biomass retention in the systems was practically not influenced by the size of particle. Biomass concentration of 1.2 g protein/L was retained in the carrier and up to 20% of the newly produced biomass was retained in the CFBRs.

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Assessment of Acid Phosphatase Production by In vitro Cultures of Atropa acuminata

Plant Tissue Culture and Biotechnology ◽

10.3329/ptcb.v26i1.29763 ◽

2016 ◽

Vol 26 (1) ◽

pp. 15-23

Author(s):

Saima Khan ◽

Meenu Katoch ◽

Sharada Mallubhotla ◽

Suphla Gupta ◽

Manju Sambyal ◽

...

Keyword(s):

Acid Phosphatase ◽

Specific Activity ◽

Polyacrylamide Gel Electrophoresis ◽

Growth Period ◽

Callus Cultures ◽

Shoot Cultures ◽

Crude Enzyme Extract ◽

Atropa Acuminata

The potential of various culture lines of Atropa acuminata were investigated for resourcing acid phosphatase (ACP) (3.1.3.2). Crude enzyme extract comprised of a mixture of four isoforms, distinguishable by polyacrylamide gel electrophoresis (PAGE) with molecular weight ranging from 39 to 215 kDa. In vitro regenerated proliferative shoots, callus and roots showed higher specific activity (2.49, 3.41, 2.91 U/mg protein, respectively) as compared to in vivo grown plants (0.71 U/mg protein). ACP activity in root cultures increased progressively up to 4.6 U/mg during the entire growth period (2 ? 24 weeks), whereas in case of shoot cultures, the specific activity escalated to 2.49 U/mg at 8 weeks, which then declined subsequently (1.95 U/mg). Similarly, callus cultures initially showed a higher phosphohydrolytic activity (3.41 U/mg protein) until 8 weeks by which period, it decreased with the passage of growth period. The present studies reveal an alternate system for resourcing of ACP from Atropa acuminata.Plant Tissue Cult. & Biotech. 26(1): 15-23, 2016 (June)

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In vitro propagation of Rosa ‘Konstancin’ (R. rugosa × R. beggeriana), a plant with high nutritional and pro-health value

Folia Horticulturae ◽

10.2478/fhort-2018-0022 ◽

2018 ◽

Vol 30 (2) ◽

pp. 259-267 ◽

Cited By ~ 2

Author(s):

Agnieszka Wojtania ◽

Bożena Matysiak

Keyword(s):

In Vitro Propagation ◽

Photosynthetic Activity ◽

Shoot Culture ◽

Bud Break ◽

Multiplication Rate ◽

Shoot Cultures ◽

Old Field ◽

Health Value

Abstract The aim of the study was to develop an efficient micropropagation system for Rosa ‘Konstancin’, an interspecific hybrid between R. rugosa and R. beggeriana, whose fruits have high pro-health value. Shoot cultures were initiated from shoot buds collected in May and August from 15-year-old field-grown Rosa ‘Konstancin’ shrubs. The effect and interaction of different concentrations of phytohormones, sucrose and iron sources on in vitro initiation, multiplication and rooting of shoots were studied. The time of collecting explants from donor plants significantly affected the initiation of shoot culture of Rosa ‘Konstancin’. Considerably higher frequency of bud break (100%) was obtained in explants isolated in August as compared to those collected at the end of May (30%). All buds developed into single shoots after 2-4 weeks of growing on the basal Murashige and Skoog medium containing 2.2 µM BAP, 0.3 µM GA3 and 88 mM of sucrose. The highest multiplication rate (4.8 shoots/explant) in a 5-week period was obtained on MS medium containing 50% of nitrogen salts, 3.1 µM BAP, 0.9 µM GA3 and 58 mM sucrose. High rooting frequency (100%) and quality of rooted plantlets was obtained on a medium containing 0.5 µM IBA, 138 µM Fe-EDDHA and 88 mM sucrose. Fe-EDDHA had a beneficial effect on the growth and photosynthetic activity of Rosa ‘Konstancin’ plantlets, which were successfully acclimatized ex vitro, with a more than 90% survival rate.

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The Stimulatory Effect of Purine-Type Cytokinins on Proliferation and Polyphenolic Compound Accumulation in Shoot Culture of Salvia viridis

Biomolecules ◽

10.3390/biom10020178 ◽

2020 ◽

Vol 10 (2) ◽

pp. 178 ◽

Cited By ~ 2

Author(s):

Izabela Grzegorczyk-Karolak ◽

Katarzyna Hnatuszko-Konka ◽

Mariola Zarzycka ◽

Łukasz Kuźma

Keyword(s):

Therapeutic Potential ◽

Shoot Culture ◽

Total Content ◽

Biomass Accumulation ◽

Hormonal Control ◽

Polyphenolic Compound ◽

Physiological Processes ◽

Best Response ◽

Alternative Sources ◽

Metabolite Content

The present study demonstrates hormonal control of Salvia viridis growth and development using four different purine-type cytokinins at different concentrations. The addition of cytokinins significantly increased biomass of cultures, proliferation rate, and, interestingly, secondary metabolite production. The best response in terms of multiplication ratio was recorded on Murashige and Skoog medium supplemented with 0.5 mg/L BPA (N-benzylotetrahydropyranyl adenine), while the greatest biomass accumulation was achieved when supplemented with 1 mg/L m-T (meta-topoline). Quantitative UPLC-DAD analysis of the hydromethanolic extract from S. viridis culture revealed the presence of 12 polyphenols: seven phenolic acids and five phenylethanoids. The highest total content of polyphenolic compounds was found in shoots cultivated on medium with 2 mg/L BPA (18.66 mg/g DW): almost twice that of control shoots. The medium was also the most optimal for the biosynthesis of rosmarinic acid, the predominant phenolic acid. However, the greater phenylethanoid accumulation was stimulated by 1 mg/L m-T: the metabolite content was above three times higher than that found in shoots grown on the control medium (8.03 mg/g DW vs. 2.37 mg/g DW). Hence, it was demonstrated that phytohormones are capable of influencing not only vital physiological processes, but therapeutic potential of plants as well. Therefore, the cytokinin-based sage cultures may be also considered as the alternative sources of bioactive compounds.

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In vitro and whole-plant magnitude and cross-resistance characterization of two imidazolinone-resistant sugarbeet (Beta vulgaris) somatic cell selections

Weed Science ◽

10.1017/s0043174500090123 ◽

1998 ◽

Vol 46 (1) ◽

pp. 24-29 ◽

Cited By ~ 8

Author(s):

Terry R. Wright ◽

Donald Penner

Keyword(s):

Somatic Cell ◽

Shoot Culture ◽

Acetolactate Synthase ◽

Cross Resistance ◽

Shoot Cultures ◽

Als Inhibitors ◽

Whole Plants ◽

Inhibitor Resistance ◽

Als Inhibitor

Acetolactate synthase (ALS)-inhibiting herbicide carryover in soil can severely affect sugarbeets grown in the year(s) following application. Two newly developed imidazolinone-resistant (IMI-R) sugarbeet somatic cell selections (Sir-13 and 93R30B) were examined for magnitude of resistance and extent of cross-resistance to other classes of ALS inhibitors and compared to a previously developed sulfonylurea-resistant (SU-R) selection, Sur. In vitro shoot culture tests indicated Sir-13 resistance was specific to imidazolinone (IMI) herbicides at approximately a 100-fold resistance compared to the sensitive control sugarbeet. Sur was 10,000-fold resistant to the sulfonylurea (SU) herbicide, chlorsulfuron, and 40-fold resistant to the triazolopyrimidine sulfonanilide (TP) herbicide, flumetsulam, but not cross-resistant to the IMI herbicides. 93R30B was selected for IMI-R from a plant homozygous for the SU-R allele,Sur, and displayed similar in vitro SU-R and TP-R as Sur, but also displayed a very high resistance to various IMI herbicides (400- to 3,600-fold). Compared to the sensitive control, Sir-13 was 300- and > 250-fold more resistant to imazethapyr and imazamox residues in soil, respectively. Response by whole plants to postemergence herbicide applications was similar to that observed in shoot cultures. Sir-13 exhibited > 100-fold resistance to imazethapyr as well as imazamox, and 93R30B showed > 250-fold resistance to both herbicides. 93R30B showed great enough resistance to imazamox to merit consideration of imazamox for use as a herbicide in these sugarbeets. Sir-13 showed a two- to threefold higher level of resistance in the homozygous vs. heterozygous state, indicating that like most ALS-inhibitor resistance traits, it was semidominantly inherited.

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