This paper presents an optimization of conditions for microshoot cultures of Nasturtium officinale R. Br. (watercress). Variants of the Murashige and Skoog (MS) medium containing different plant growth regulators (PGRs): cytokinins—BA (6-benzyladenine), 2iP (6-γ,γ-dimethylallylaminopurine), KIN (kinetin), Zea (zeatin), and auxins—IAA (3-indoleacetic acid), IBA (indole-3-butyric acid), 2,4-d (2,4-dichlorophenoxyacetic acid), IPA (indole-3-pyruvic acid), NAA (naphthalene-1-acetic acid), total 27 MS variants, were tested in agar and agitated cultures. Growth cycles were tested for 10, 20, or 30 days in the agar cultures, and 10 or 20 days in the agitated cultures. Glucosinolate and phenolic acid production, total phenolic content and antioxidant potential were evaluated. The total amounts of glucosinolates ranged from 100.23 to 194.77 mg/100 g dry weight of biomass (DW) in agar cultures, and from 78.09 to 182.80 mg/100 g DW in agitated cultures. The total phenolic acid content varied from 15.89 to 237.52 mg/100 g DW for the agar cultures, and from 70.80 to 236.74 mg/100 g DW for the agitated cultures. Extracts of the cultured biomass contained higher total amounts of phenolic acids, lower total amounts of glucosinolates, a higher total phenolic content and similar antioxidant potentials compared to plant material. The analyses performed confirmed for the first time the explicit influence on secondary metabolite production and on the antioxidant potential. The significance was statistically estimated in a complex manner.Nasturtium officinale R. Br. (Robert Brown, described earlier by Linnaeus as Sisymbrium nasturtium-aquaticum), called watercress, belongs to the family Brassicaceae. It is a rare perennial, aquatic or semi-aquatic herb with characteristic creeping or floating stems. In its natural habitat it colonizes gently flowing and shallow streams. N. officinale is native to Europe, North Africa and Asia [1]. In Europe, this species has been classified by the International Union for Conservation of Nature (IUCN) Red List of Threatened Species, as a plant of least concern. However, the classification of N. officinale is different in each European country; for example, it is considered an extinct or probably extinct plant in Estonia, endangered in Austria and Sweden, and partly endangered in Poland [2]. The monograph of the N. officinale herb is present in German Commission E Monographs (Phyto-Therapy) [3], and it also has an important position in German Commission D Monographs [4] for homeopathic medicines. Additionally, the European Food Safety Authority (EFSA) has classified this species as a safe vegetable in the group of “leaf vegetables, herbs and edible flowers” [5]. Its medicinal effects were used in the traditional medicine of Iran, Azerbaijan, Morocco and Mauritius to treat hyperglycemia, hypertension, asthma and cough [6,7,8]. Current studies have confirmed a wide spectrum of biological activity determined by a rich chemical composition, such as anticancer, antioxidant, antimicrobial, anti-inflammatory, antipsoriatic and cardioprotective [9,10,11,12]. The main groups of compounds found in the herb are: glucosinolates, isothiocyanates, polyphenols (flavonoids, phenolic acids, proanthocyanidins), terpenoids (including carotenoids), vitamins (B1, B2, B3, B6, E, C) and bioelements [13,14,15,16,17]. N. officinale is also used in the cosmetic industry as an antioxidant, anti-ageing, skin-lightening and anti-acne agent [18]. N. officinale is also increasingly popular in modern cuisine. Currently, it is added to European dishes such as salads and soups because of its rich composition and numerous scientifically proven properties [1]. Moreover, it has aproven valuable phytoremediation capacity. Current studies have confirmed its ability to purify water and remove copper (Cu), nickel (Ni), zinc (Zn) [19], cadmium (Cd), cobalt (Co), chromium (Cr) [20,21], arsenic (As) [22] and uranium (U) [23] from soil.The first experiments involved basal studies of MS media with the addition of one of the following cytokinins: BA, 2iP, KIN, Zea, at a concentration of 1 mg/L each. The control medium, without any addition of PGRs, was used for comparison. In the experiment shown, small increases of biomass and the number of microshoots in the in vitro cultures growing in the medium with the addition of 1 mg/L of one cytokinin only were observed. Variants containing 1 mg/L Zea and 1 mg/L BA and the control medium were characterized by a higher number of microshoots and the dark green color of the shoots and leaves. The Gi values obtained ranged from 0.05 to 3.26. The maximum values were for microshoots growing for 20 days on variant 0 (control) of the medium. A high Gi (2.55) was also obtained for in vitro cultures growing on the MS medium variant containing 1 mg/L BA, after 30 days (Figure S1, Table S2). Good results were obtained with the use of BA as a cytokinin and NAA as an auxin in different concentrations: 2 and 1, 1 and 2, 1 and 1 (mg/L), respectively. Microshoots growing on all these variants were characterized by a large number of microshoots and the dark green color of the leaves. The highest Gi (5.05) was observed after 30 days on the MS medium variant containing 1 mg/L BA and 1 mg/L NAA (Figure S1, Table 1 and Table S2)

Question

This paper presents an optimization of conditions for microshoot cultures of Nasturtium officinale R. Br. (watercress). Variants of the Murashige and Skoog (MS) medium containing different plant growth regulators (PGRs): cytokinins—BA (6-benzyladenine), 2iP (6-γ,γ-dimethylallylaminopurine), KIN (kinetin), Zea (zeatin), and auxins—IAA (3-indoleacetic acid), IBA (indole-3-butyric acid), 2,4-d (2,4-dichlorophenoxyacetic acid), IPA (indole-3-pyruvic acid), NAA (naphthalene-1-acetic acid), total 27 MS variants, were tested in agar and agitated cultures. Growth cycles were tested for 10, 20, or 30 days in the agar cultures, and 10 or 20 days in the agitated cultures. Glucosinolate and phenolic acid production, total phenolic content and antioxidant potential were evaluated. The total amounts of glucosinolates ranged from 100.23 to 194.77 mg/100 g dry weight of biomass (DW) in agar cultures, and from 78.09 to 182.80 mg/100 g DW in agitated cultures. The total phenolic acid content varied from 15.89 to 237.52 mg/100 g DW for the agar cultures, and from 70.80 to 236.74 mg/100 g DW for the agitated cultures. Extracts of the cultured biomass contained higher total amounts of phenolic acids, lower total amounts of glucosinolates, a higher total phenolic content and similar antioxidant potentials compared to plant material. The analyses performed confirmed for the first time the explicit influence on secondary metabolite production and on the antioxidant potential. The significance was statistically estimated in a complex manner.Nasturtium officinale R. Br. (Robert Brown, described earlier by Linnaeus as Sisymbrium nasturtium-aquaticum), called watercress, belongs to the family Brassicaceae. It is a rare perennial, aquatic or semi-aquatic herb with characteristic creeping or floating stems. In its natural habitat it colonizes gently flowing and shallow streams. N. officinale is native to Europe, North Africa and Asia [1]. In Europe, this species has been classified by the International Union for Conservation of Nature (IUCN) Red List of Threatened Species, as a plant of least concern. However, the classification of N. officinale is different in each European country; for example, it is considered an extinct or probably extinct plant in Estonia, endangered in Austria and Sweden, and partly endangered in Poland [2]. The monograph of the N. officinale herb is present in German Commission E Monographs (Phyto-Therapy) [3], and it also has an important position in German Commission D Monographs [4] for homeopathic medicines. Additionally, the European Food Safety Authority (EFSA) has classified this species as a safe vegetable in the group of “leaf vegetables, herbs and edible flowers” [5]. Its medicinal effects were used in the traditional medicine of Iran, Azerbaijan, Morocco and Mauritius to treat hyperglycemia, hypertension, asthma and cough [6,7,8]. Current studies have confirmed a wide spectrum of biological activity determined by a rich chemical composition, such as anticancer, antioxidant, antimicrobial, anti-inflammatory, antipsoriatic and cardioprotective [9,10,11,12]. The main groups of compounds found in the herb are: glucosinolates, isothiocyanates, polyphenols (flavonoids, phenolic acids, proanthocyanidins), terpenoids (including carotenoids), vitamins (B1, B2, B3, B6, E, C) and bioelements [13,14,15,16,17]. N. officinale is also used in the cosmetic industry as an antioxidant, anti-ageing, skin-lightening and anti-acne agent [18]. N. officinale is also increasingly popular in modern cuisine. Currently, it is added to European dishes such as salads and soups because of its rich composition and numerous scientifically proven properties [1]. Moreover, it has aproven valuable phytoremediation capacity. Current studies have confirmed its ability to purify water and remove copper (Cu), nickel (Ni), zinc (Zn) [19], cadmium (Cd), cobalt (Co), chromium (Cr) [20,21], arsenic (As) [22] and uranium (U) [23] from soil.The first experiments involved basal studies of MS media with the addition of one of the following cytokinins: BA, 2iP, KIN, Zea, at a concentration of 1 mg/L each. The control medium, without any addition of PGRs, was used for comparison. In the experiment shown, small increases of biomass and the number of microshoots in the in vitro cultures growing in the medium with the addition of 1 mg/L of one cytokinin only were observed. Variants containing 1 mg/L Zea and 1 mg/L BA and the control medium were characterized by a higher number of microshoots and the dark green color of the shoots and leaves. The Gi values obtained ranged from 0.05 to 3.26. The maximum values were for microshoots growing for 20 days on variant 0 (control) of the medium. A high Gi (2.55) was also obtained for in vitro cultures growing on the MS medium variant containing 1 mg/L BA, after 30 days (Figure S1, Table S2). Good results were obtained with the use of BA as a cytokinin and NAA as an auxin in different concentrations: 2 and 1, 1 and 2, 1 and 1 (mg/L), respectively. Microshoots growing on all these variants were characterized by a large number of microshoots and the dark green color of the leaves. The highest Gi (5.05) was observed after 30 days on the MS medium variant containing 1 mg/L BA and 1 mg/L NAA (Figure S1, Table 1 and Table S2)

write only the main points

Answer 1

1. Optimal conditions for microshoot cultures of Nasturtium officinale R. Br. were determined through testing different variants of the MS medium containing various plant growth regulators in both agar and agitated cultures.

2. Glucosinolate and phenolic acid production, total phenolic content, and antioxidant potential were evaluated in the cultured biomass.

3. The total amounts of glucosinolates ranged from 100.23 to 194.77 mg/100 g dry weight in agar cultures and from 78.09 to 182.80 mg/100 g dry weight in agitated cultures.

4. The total phenolic acid content varied from 15.89 to 237.52 mg/100 g dry weight in agar cultures and from 70.80 to 236.74 mg/100 g dry weight in agitated cultures.

5. Extracts of the cultured biomass showed higher total amounts of phenolic acids, lower total amounts of glucosinolates, higher total phenolic content, and similar antioxidant potentials compared to plant material.

6. The study confirmed the influence of different plant growth regulators on secondary metabolite production and antioxidant potential in N. officinale microshoot cultures.

7. N. officinale is a valuable herb with a rich chemical composition and numerous scientifically proven properties, including anticancer, antioxidant, antimicrobial, anti-inflammatory, and cardioprotective effects.

8. The herb is also used in traditional medicine, cosmetics, phytoremediation, and modern cuisine, making it a versatile and beneficial plant species.