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2 pages, 178 KiB

Open AccessEditorial

Plant Tissue Culture and Secondary Metabolites Production

by Kalina Danova and Laura Pistelli

Plants 2022, 11(23), 3312; https://doi.org/10.3390/plants11233312 - 30 Nov 2022

Cited by 8 | Viewed by 5622

Abstract

Plants have developed a complex biochemical system for interacting and coping with dynamic environmental challenges throughout their whole life [...] Full article

(This article belongs to the Special Issue Plant Tissue Culture and Secondary Metabolites Production)

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15 pages, 1619 KiB

Open AccessArticle

Halophyte Artemisia caerulescens L.: Metabolites from In Vitro Shoots and Wild Plants

by Ylenia Pieracci, Martina Vento, Luisa Pistelli, Tiziana Lombardi and Laura Pistelli

Plants 2022, 11(8), 1081; https://doi.org/10.3390/plants11081081 - 15 Apr 2022

Cited by 6 | Viewed by 2360

Abstract

Halophyte plants are potential resources to deal with the increasing soil salinity determined by climatic change. In this context, the present study aimed to investigate the germplasm conservation of Artemisia caerulescens collected in the San Rossore Estate (Pisa, Italy) through in vitro culture, [...] Read more.

Halophyte plants are potential resources to deal with the increasing soil salinity determined by climatic change. In this context, the present study aimed to investigate the germplasm conservation of Artemisia caerulescens collected in the San Rossore Estate (Pisa, Italy) through in vitro culture, biochemical properties, and the phytochemical composition of the volatile fraction of both in vitro shoots and different organs of wild plants (leaves, young and ripe inflorescences). The best medium tested for the shoot proliferation was MS, with the addition of 1 μM BA. Total polyphenol content and antioxidant activity were noticeable in both the inflorescences, while leaves and in vitro shoots showed lower amounts. Concerning the phytochemical investigation, the headspaces (HSs) and the essential oils (EOs) were characterized by oxygenated monoterpenes as the main chemical class of compounds in all samples, and with α- and β-thujone as the major constituents. However, the EOs were characterized by noticeable percentages of phenylpropanoids (23.6–28.8%), with brevifolin as the unique compound, which was not detected in the spontaneous volatile emissions of the same parts of the wild plant. Good amounts of EOs were obtained from different organs of the wild plant, comprising between 0.17% and 0.41% of the young and ripe inflorescences, respectively. Full article

(This article belongs to the Special Issue Plant Tissue Culture and Secondary Metabolites Production)

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17 pages, 4771 KiB

Open AccessArticle

Optimization of Biomass Accumulation and Production of Phenolic Compounds in Callus Cultures of Rhodiola rosea L. Using Design of Experiments

by Anna A. Erst, Anastasia A. Petruk, Andrey S. Erst, Denis A. Krivenko, Nadezhda V. Filinova, Svetlana Y. Maltseva, Maxim S. Kulikovskiy and Evgeny V. Banaev

Plants 2022, 11(1), 124; https://doi.org/10.3390/plants11010124 - 2 Jan 2022

Cited by 9 | Viewed by 3238

Abstract

Rhodiola rosea L. is a valuable medicinal plant with adaptogenic, neuroprotective, antitumor, cardioprotective, and antidepressant effects. In this study, design of experiments methodology was employed to analyze and optimize the interacting effects of mineral compounds (concentration of NO₃⁻ and the ratio [...] Read more.

Rhodiola rosea L. is a valuable medicinal plant with adaptogenic, neuroprotective, antitumor, cardioprotective, and antidepressant effects. In this study, design of experiments methodology was employed to analyze and optimize the interacting effects of mineral compounds (concentration of NO₃⁻ and the ratio of NH₄⁺ to K⁺) and two plant growth regulators [total 6-benzylaminopurine (BAP) and α-naphthylacetic acid (NAA) concentration and the ratio of BAP to NAA] on the growth and the production of total phenolic compounds (TPCs) in R. rosea calluses. The overall effect of the model was highly significant (p < 0.0001), indicating that NH₄⁺, K⁺, NO₃⁻, BAP, and NAA significantly affected growth. The best callus growth (703%) and the highest production of TPCs (75.17 mg/g) were achieved at an NH₄⁺/K⁺ ratio of 0.33 and BAP/NAA of 0.33, provided that the concentration of plant growth regulators was 30 μM and that of NO₃⁻ was ≤40 mM. According to high-performance liquid chromatography analyses of aerial parts (leaves and stems), in vitro seedlings and callus cultures of R. rosea contain no detectable rosarin, rosavin, rosin, and cinnamyl alcohol. This is the first report on the creation of an experiment for the significant improvement of biomass accumulation and TPC production in callus cultures of R. rosea. Full article

(This article belongs to the Special Issue Plant Tissue Culture and Secondary Metabolites Production)

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16 pages, 2583 KiB

Open AccessArticle

Genetic Stability, Phenolic, Flavonoid, Ferulic Acid Contents, and Antioxidant Activity of Micropropagated Lycium schweinfurthii Plants

by Diaa Mamdouh, Hany A. M. Mahgoub, Ahmed M. M. Gabr, Emad A. Ewais and Iryna Smetanska

Plants 2021, 10(10), 2089; https://doi.org/10.3390/plants10102089 - 1 Oct 2021

Cited by 15 | Viewed by 3448

Abstract

Lycium schweinfurthii is a Mediterranean wild shrub rich in plant secondary metabolites. In vitro propagation of this plant may support the production of valuable dietary supplements for humanity, introduction of it to the world market, and opportunities for further studies. The presented study [...] Read more.

Lycium schweinfurthii is a Mediterranean wild shrub rich in plant secondary metabolites. In vitro propagation of this plant may support the production of valuable dietary supplements for humanity, introduction of it to the world market, and opportunities for further studies. The presented study aimed to introduce an efficient and reproducible protocol for in vitro micropropagation of L. schweinfurthii and assess the genetic stability of micropropagated plants (MiPs) as well as to estimate phenolic, flavonoid, ferulic acid contents, and the antioxidant activity in leaves of micropropagated plants. Two DNA-based techniques, random amplified polymorphic DNA (RAPD) and inter-simple sequence repeats (ISSR), and one biochemical technique, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), were used to assess the genetic stability in MiPs. Spectrophotometric analysis was performed to estimate total phenolic and flavonoid contents and antioxidant activity of MiPs leaves, while ferulic acid content was estimated using high-performance thin-layer chromatography (HPTLC). Sufficient shoot proliferation was achieved at MS (Murashige and Skoog) medium supplemented with 0.4 mg L⁻¹ kinetin and rooted successfully on half-strength MS medium fortified with 0.4 mg L⁻¹ Indole-3-butyric acid (IBA). The Jaccard’s similarity coefficients detected in MiPs reached 52%, 55%, and 82% in the RAPD, ISSR, and SDS-PAGE analyses, respectively. In the dried leaves of MiPs, the phenolic, flavonoid, and ferulic acid contents of 11.53 mg gallic acid equivalent, 12.99 mg catechin equivalent, and 45.52 mg were estimated per gram, respectively. However, an IC₅₀ of 0.43, and 1.99 mg mL⁻¹ of MiP dried leaves’ methanolic extract was required to scavenge half of the DPPH, and ABTS free radicals, respectively. The study presented a successful protocol for in vitro propagation of a valued promising plant source of phenolic compounds. Full article

(This article belongs to the Special Issue Plant Tissue Culture and Secondary Metabolites Production)

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14 pages, 9475 KiB

Open AccessArticle

Repression of Carotenoid Accumulation by Nitrogen and NH₄⁺ Supply in Carrot Callus Cells In Vitro

by Tomasz Oleszkiewicz, Michał Kruczek and Rafal Baranski

Plants 2021, 10(9), 1813; https://doi.org/10.3390/plants10091813 - 31 Aug 2021

Cited by 9 | Viewed by 3336

Abstract

The effect of mineral nutrition on the accumulation of the main health beneficial compounds in carrots, the carotenoid pigments, remains ambiguous; here, a model-based approach was applied to reveal which compounds are responsible for the variation in carotenoid content in carrot cells in [...] Read more.

The effect of mineral nutrition on the accumulation of the main health beneficial compounds in carrots, the carotenoid pigments, remains ambiguous; here, a model-based approach was applied to reveal which compounds are responsible for the variation in carotenoid content in carrot cells in vitro. For this purpose, carotenoid-rich callus was cultured on either BI (modified Gamborg B5) or R (modified Murashige and Skoog MS) mineral media or on modified media obtained by exchanging compounds between BI and R. Callus growing on the BI medium had abundant carotene crystals in the cells and a dark orange color in contrast to pale orange callus with sparse crystals on the R medium. The carotenoid content, determined by HPLC and spectrophotometrically after two months of culture, was 5.3 higher on the BI medium. The replacement of media components revealed that only the N concentration and the NO₃:NH₄ ratio affected carotenoid accumulation. Either the increase of N amount above 27 mM or decrease of NO₃:NH₄ ratio below 12 resulted in the repression of carotenoid accumulation. An adverse effect of the increased NH₄⁺ level on callus growth was additionally found. Somatic embryos were formed regardless of the level of N supplied. Changes to other media components, i.e., macroelements other than N, microelements, vitamins, growth regulators, and sucrose had no effect on callus growth and carotenoid accumulation. The results obtained from this model system expand the range of factors, such as N availability, composition of N salts, and ratio of nitrate to ammonium N form, that may affect the regulation of carotenoid metabolism. Full article

(This article belongs to the Special Issue Plant Tissue Culture and Secondary Metabolites Production)

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14 pages, 2502 KiB

Open AccessArticle

In Vitro Propagation Method for Production of Phenolic-Rich Planting Material of Culinary Rhubarb ‘Malinowy’

by Agnieszka Wojtania and Monika Mieszczakowska-Frąc

Plants 2021, 10(9), 1768; https://doi.org/10.3390/plants10091768 - 25 Aug 2021

Cited by 9 | Viewed by 2559

Abstract

Culinary rhubarb is a popular vegetable crop, valued for its long, thickened stalks, very rich in different natural bioactive ingredients. Tissue cultures are a useful tool for vegetative propagation of virus-free rhubarb plants and rapid multiplication of valuable selected genotypes. The aim of [...] Read more.

Culinary rhubarb is a popular vegetable crop, valued for its long, thickened stalks, very rich in different natural bioactive ingredients. Tissue cultures are a useful tool for vegetative propagation of virus-free rhubarb plants and rapid multiplication of valuable selected genotypes. The aim of this study was to develop an effective method for in vitro propagation of selected genotypes of Polish rhubarb ‘Malinowy’ characterized by high yield and straight, thick and intensive red stalks. Identification and quantification of anthocyanins and soluble sugars by the HPLC method in shoot cultures and ex vitro established plantlets were also performed. Shoot cultures were established from axillary buds isolated from dormant, eight-year-old rhizomes. Effective shoot multiplication of rhubarb ‘Malinowy’ was obtained in the presence of 6.6 µM benzylaminopurine or 12.4 µM meta-topolin. Both cytokinins stimulated shoot formation in a manner that depended on sucrose concentration. Increasing the sucrose concentration from 59 to 175 mM decreased the production of shoots and outgrowth of leaves by 3-fold but enhanced shoot length, single shoot mass and callus formation at the base of shoots. This coincided with increased accumulation of soluble sugars (fructose, glucose) and anthocyanins-cyanidin-3-O-rutinoside (max. 208.2 mg·100 g⁻¹ DM) and cyanidin-3-O-glucoside (max. 47.7 mg·100 g⁻¹ DM). The highest rooting frequency (94.9%) and further successful ex vitro establishment (100%) were observed for shoots that were earlier rooted in vitro in the presence of 4.9 µM indole-3-butyric acid. Our results indicated that anthocyanin contents in leaf petioles were influenced by developmental stage. Under in vitro conditions, it is possible to elicit those pigments by sucrose at high concentration and meta-topolin. Full article

(This article belongs to the Special Issue Plant Tissue Culture and Secondary Metabolites Production)

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21 pages, 4661 KiB

Open AccessArticle

Gentianella lutescens subsp. carpatica J. Holub.: Shoot Propagation In Vitro and Effect of Sucrose and Elicitors on Xanthones Production

by Dijana Krstić-Milošević, Nevena Banjac, Teodora Janković, Dragan Vinterhalter and Branka Vinterhalter

Plants 2021, 10(8), 1651; https://doi.org/10.3390/plants10081651 - 11 Aug 2021

Cited by 5 | Viewed by 2208

Abstract

In vitro shoot culture of the endangered medicinal plant Gentianella lutescens was established from epicotyl explants cultured on MS basal medium with 0.2 mg L⁻¹ 6-benzylaminopurine (BA) and evaluated for xanthones content for the first time. Five shoot lines were obtained and [...] Read more.

In vitro shoot culture of the endangered medicinal plant Gentianella lutescens was established from epicotyl explants cultured on MS basal medium with 0.2 mg L⁻¹ 6-benzylaminopurine (BA) and evaluated for xanthones content for the first time. Five shoot lines were obtained and no significant variations in multiplication rate, shoot elongation, and xanthones profile were found among them. The highest rooting rate (33.3%) was achieved by shoots treated for 2 days with 5 mg L⁻¹ indole-3-butyric acid (IBA) followed by cultivation in liquid PGR-free ½ MS medium for 60 days. HPLC analysis revealed the lower content of xanthones—mangiferin, bellidifolin, demethylbellidifolin, demethylbellidifolin-8-O-glucoside and bellidifolin-8-O-glucoside—in in vitro cultured shoots compared to wild growing plants. The increasing concentration of sucrose, sorbitol and abiotic elicitors salicylic acid (SA), jasmonic acid (JA) and methyl jasmonate (MeJA) altered shoot growth and xanthone production. Sucrose and sorbitol applied at the highest concentration of 233.6 mM increased dry matter percentage, while SA at 100 μM promoted shoot growth 2-fold. The increased sucrose concentration enhanced accumulation of xanthones in shoot cultures 2–3-fold compared to the control shoots. Elicitors at 100–300 μM increased the accumulation of mangiferin, demethylbellidifolin-8-O-glucoside, and bellidifolin-8-O-glucoside almost equally, while MeJA at the highest concentration of 500 μM enhanced amount of aglycones demethylbellidifolin and bellidifolin 7-fold compared to the control. The obtained results facilitate conservation of G. lutescens and pave the way for further research on large-scale shoot propagation and production of pharmacologically active xanthones. Full article

(This article belongs to the Special Issue Plant Tissue Culture and Secondary Metabolites Production)

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14 pages, 2183 KiB

Open AccessArticle

Application of Plant Growth Regulators Modulates the Profile of Chlorogenic Acids in Cultured Bidens pilosa Cells

by Anza-Tshilidzi Ramabulana, Paul A. Steenkamp, Ntakadzeni E. Madala and Ian A. Dubery

Plants 2021, 10(3), 437; https://doi.org/10.3390/plants10030437 - 25 Feb 2021

Cited by 14 | Viewed by 4061

Abstract

Plant cell culture offers an alternative to whole plants for the production of biologically important specialised metabolites. In cultured plant cells, manipulation by auxin and cytokinin plant growth regulators (PGRs) may lead to in vitro organogenesis and metabolome changes. In this study, six [...] Read more.

Plant cell culture offers an alternative to whole plants for the production of biologically important specialised metabolites. In cultured plant cells, manipulation by auxin and cytokinin plant growth regulators (PGRs) may lead to in vitro organogenesis and metabolome changes. In this study, six different combination ratios of 2,4-dichlorophenoxyacetic acid (2,4-D) and benzylaminopurine (BAP) were investigated with the aim to induce indirect organogenesis from Bidens pilosa callus and to investigate the associated induced changes in the metabolomes of these calli. Phenotypic appearance of the calli and total phenolic contents of hydromethanolic extracts indicated underlying biochemical differences that were investigated using untargeted metabolomics, based on ultra-high-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UHPLC–qTOF–MS), combined with multivariate data analysis. The concentration and combination ratios of PGRs were shown to induce differential metabolic responses and, thus, distinct metabolomic profiles, dominated by chlorogenic acids consisting of caffeoyl- and feruloyl-derivatives of quinic acid. Although organogenesis was not achieved, the results demonstrate that exogenous application PGRs can be used to manipulate the metabolome of B. pilosa for in vitro production of specialised metabolites with purported pharmacological properties. Full article

(This article belongs to the Special Issue Plant Tissue Culture and Secondary Metabolites Production)

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