Accumulation of valuable secondary metabolites: phenolic acids and flavonoids in different in vitro systems of shoot cultures of the endangered plant species—Eryngium alpinum L.

Abstract

In vitro cultures give the opportunity to perform the phytochemical studies on the protected species without harvesting the plant material from the natural environment. Shoots of Eryngium alpinum L. were multiplied on Murashige and Skoog (MS) medium in various systems, namely on the solid media and in two liquid cultures-stationary and agitated, as well as via regeneration from callus. The biomass increments were closely correlated with the number of shoots arising from one explant, which was connected with the supplementation of the culture media with the studied plant growth regulators. The methanolic extracts from shoots grown in the tested systems were subjected to phenolic acids and flavonoids qualitative and quantitative analysis. Biomass from in vitro shoot cultures accumulated from 19.59 to 32.95 times more phenolic acids [the total content ranged from 272.52 to 458.38 mg/100 g dry weight (DW)] and from 3.02 to 4.43 times more flavonoids (the total content ranged from 100.03 to 146.98 mg/100 g DW), depending on the culture system, than the extracts from basal leaves from the intact plant (13.91 and 33.16 mg/100 g DW, respectively). The phenolics present in shoot cultures include seven phenolic acids-3,4-dihydroxyphenylacetic, caftaric, caffeic, neochlorogenic, chlorogenic, isochlorogenic, and rosmarinic acids, and three flavonoids-isoquercetin, quercitrin and robinin. The best system for shoot proliferation resulting in the highest biomass growth and phenolic acids and flavonoids accumulation was solid culture on MS medium with BAP, IAA, and GA 3 (each 1.0 mg/l). The aim of this work was to check the effect of various culture systems (stationary and agitated, on solidified and in liquid media) on the production of phenolic compounds in E. alpinum shoots cultured in vitro. Key messageThis is the first report on phenolic acids and flavonoids estimation in Eryngium alpinum in vitro biomass from different culture systems.Keywords Alpine eryngo · Shoot multiplication · Phenolic acids · Flavonoids · HPLC-DAD analysis 1995). Some of the individuals are threatened by humansthe plant is collected for ornamental and commercial use and

Phytochemical Screening and Acanthamoebic Activity of Shoots from in Vitro Cultures and in Vivo Plants of Eryngium alpinum L.—The Endangered and Protected Species

“…This study indicated the alternative method for effective and rapid shoot multiplication of E. alpinum. However, the increase in the concentration of BAP did not provide the highest biotechnological parameters compared to our previous studies [6,7]. In the case of E. planum, the highest mean number of shoots developed via axillary buds was 15.58 ± 0.54-17.10 ± 0.60 shoots per explant, depending on the culture media: MS + BAP 1.0 mg/L + IAA 1.0 mg/L or MS + BAP 1.0 mg/L + IAA 0.1 mg/L [14].…”

“…One of the major groups of compounds were hydroxycinnamic acid derivates, which include conjugates of coumaric, caffeic and ferulic acid with the hexose (neutral losses −162.0834, C 6 H 10 O 5 ) and the quinic acid (characteristic fragment m/z 191.0195, C 7 H 11 O 6 − ). Three conjugates of caffeic acid and quinic acid were annotated in the sample namely neochlorogenic acid (5-caffeoylquinic acid, RT = 5.57 min), chlorogenic acid (3-caffeoylquinic acid, RT = 6.06), and isochlorogenic acid (5Z-caffeoylquinic acid, RT = 7.02 min); they were previously described by Kikowska et al [6,7]. The pseudo-molecular ion m/z 353.08743 corresponded with the formula C 16 Coumarins such as umbeliferone, scopoletin, 7-methoxycoumarin, and dihydroxycoumarin were recognized in our sample basing on the exact mass and the fragmentation pattern and were previously described for the different Eryngium species and the Apiaceae family [18], however, for the first time they were recognized in E. alpinum.…”

“…Due to the unavailability of the plant material, little research on this taxon was carried out. Only a few papers indicated the presence of phenolic acids, flavonoids and the essential oil in the organs of in vivo plants [3][4][5][6][7]. The identification of flavonoids, namely quercetin and kaempferol, in leaves of alpine eryngo was described by Crowden et al [3].…”

Chemical Composition of Essential Oils from Rare and Endangered Species—Eryngium maritimum L. and E. alpinum L.

“…The explants of both the species were placed in a Erlenmeyer flask with 50 mL of the solidified MS medium (Murashige and Skoog [39]) with plant growth regulators-benzylaminopurine (BAP; Sigma-Aldrich, Saint Louis, MO, USA), indole-3-acetic acid (IAA; Sigma-Aldrich, Saint Louis, MO, USA), and gibberellic acid (GA 3 ; Sigma-Aldrich, Saint Louis, MO, USA), each at the concentration of 1.0 mg/L [23,40]. The culture vessels were placed in a growth chamber (21 ± 2 • C; with a 16 h light/8 h dark photoperiod; 55 µmol/m 2 •s light) and subcultured every 5 weeks.…”

Powerful Plant Antioxidants: A New Biosustainable Approach to the Production of Rosmarinic Acid

“…These in vitro techniques have been widely used for the production of several SMs, notably taxol [ 31 , 32 , 33 ], podophyllotoxin [ 34 , 35 , 36 ], withanolides [ 37 ], centellosides [ 38 , 39 ], and rosmarinic acid (RA) [ 40 , 41 , 42 , 43 ], as well as for mass propagation [ 44 ], in vitro cloning [ 45 , 46 , 47 ], and polyploidization [ 48 , 49 ]. Many RA-producing biotechnological platforms have been established, based on shoots [ 50 , 51 , 52 ], callus [ 53 ], cells [ 41 , 42 , 54 ], and hairy root cultures [ 55 , 56 , 57 , 58 ] of numerous species of the Boraginaceae, Anthocerotaceae, and Lamiaceae.…”