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Plant Biotechnology Applications in Secondary Metabolite Production
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Plant Biotechnology Applications in Secondary Metabolite Production

A special issue of Plants (ISSN 2223-7747). This special issue belongs to the section "Plant Physiology and Metabolism".

Deadline for manuscript submissions: closed (20 January 2024) | Viewed by 63265

Special Issue Editor

Dr. Mauro Commisso

E-Mail Website
Guest Editor
Department of Biotechnology, University of Verona, Strada Le Grazie 15, 37134 Verona, Italy
Interests: untargeted metabolomics; targeted metabolomics; general botany; plant cell biology; in vitro plant cell cultures; phytochemicals; fruit quality; plant metabolism and chemometrics

Special Issue Information

Dear Colleagues, 

Plants cells are incredible sources of small molecules, named secondary/specialized metabolites, usually classified according to their chemical structure in different groups, including phenolic compounds, organic acids, glucosinolates, cyanogenic glucosides, amines, non-protein amino acids, peptides, alkaloids, and terpenes. Many secondary metabolites are commercially interesting, and they are currently exploited in several fields, such as in the production of ordinary food, novel food, cosmetics, agrochemicals, fine chemicals, biopharmaceuticals, and nutraceuticals.

Plant biotechnology exploits genetic, chemical, and biological knowledges and approaches to adapting plants for specific human necessities. For example, the characterization of the secondary metabolome of new plant varieties might provide information for the production of novel food rich in antioxidants or with particular flavors. Again, in vitro plant tissues and in vitro plant cell cultures might be precious and “green” sources of valuable secondary metabolites, for example, for the production of biopharmaceuticals.

This Special Issue of Plants focuses on plant secondary metabolites and the use of plant biotechnology applications related to the production of molecules of interests.

In this sense, reviews and research papers or short communications on topics related to the investigations and characterizations of the secondary metabolome of whole plants (plant organs and tissues), new plant varieties, poorly investigated plant species, in vitro plant tissue cultures, and in vitro plant cell cultures are welcome, as well as strategies to improve the accumulation of metabolites of interests.

Dr. Mauro Commisso
Guest Editor

Manuscript Submission Information

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Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Plants is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2700 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • secondary metabolites
  • specialized metabolites
  • plant cell cultures
  • plant tissue cultures
  • natural products
  • phytochemicals

Published Papers (19 papers)

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Research

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15 pages, 4487 KiB  
Article
Metabolomics Reveals Rubiadin Accumulation and the Effects of Methyl Jasmonate Elicitation in Damnacanthus major Calli
by Hyejin Hyeon, Eun Bi Jang, Sung Chun Kim, Seon-A Yoon, Boram Go, Jong-Du Lee, Ho Bong Hyun and Young-Min Ham
Plants 2024, 13(2), 167; https://doi.org/10.3390/plants13020167 - 08 Jan 2024
Viewed by 761
Abstract
Callus suspension techniques have been considered attractive for improving bioactive metabolite productivity; methyl jasmonate (MeJA) is a widely used elicitor for stimulating synthetic pathways. In this study, a multivariate analysis-based metabolomics approach was employed to investigate the primary and specialized metabolites in the [...] Read more.
Callus suspension techniques have been considered attractive for improving bioactive metabolite productivity; methyl jasmonate (MeJA) is a widely used elicitor for stimulating synthetic pathways. In this study, a multivariate analysis-based metabolomics approach was employed to investigate the primary and specialized metabolites in the leaves, unelicited calli, and 100 or 200 μM MeJA elicited calli of Damnacanthus major. Rubiadin, a powerful anthraquinone with various therapeutic properties, was only identified in D. major calli, accumulating in a MeJA elicitation concentration-dependent manner. Callus cultures also contained high levels of amino acids, sugars, and phenolic compounds, indicating energy metabolism and metabolic adaptation responses for proliferation and stabilization. Regarding MeJA application, elicited calli contained higher amounts of quinic acid, kaempferol, and glucose with lower amounts of sucrose and raffinose than those in the unelicited control, which were closely related to protective mechanisms against MeJA. Moreover, excessive elicitation increased the asparagine, fructose, and raffinose levels and decreased the glucose and sucrose levels, which was ascribed to increased activation of the aminoacyl-tRNA biosynthesis pathway and wider utilization of glucose than of fructose after sucrose degradation. These results will be useful for optimizing plant cell culture techniques to achieve high production rates for valuable specialized metabolites. Full article
(This article belongs to the Special Issue Plant Biotechnology Applications in Secondary Metabolite Production)
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20 pages, 2480 KiB  
Article
Exploring the Phytochemical Composition and Biological Potential of Balkan Endemic Species Stachys scardica Griseb
by Desislava I. Mantovska, Miroslava K. Zhiponova, Detelina Petrova, Kalina Alipieva, Georgi Bonchev, Irina Boycheva, Yana Evstatieva, Dilyana Nikolova, Ivanka Tsacheva, Svetlana Simova and Zhenya P. Yordanova
Plants 2024, 13(1), 30; https://doi.org/10.3390/plants13010030 - 21 Dec 2023
Cited by 1 | Viewed by 931
Abstract
Stachys scardica Griseb. is a Balkan endemic species listed in The Red Data Book of Bulgaria with the conservation status “endangered”. Successful micropropagation was achieved on MS medium supplemented with 1.5 mg/L benzyladenine (BA), followed by a subsequent ex vitro adaptation in an [...] Read more.
Stachys scardica Griseb. is a Balkan endemic species listed in The Red Data Book of Bulgaria with the conservation status “endangered”. Successful micropropagation was achieved on MS medium supplemented with 1.5 mg/L benzyladenine (BA), followed by a subsequent ex vitro adaptation in an experimental field resulting in 92% regenerated plants. Using nuclear magnetic resonance (NMR), phenylethanoid glycosides (verbascoside, leucosceptoside A), phenolic acids (chlorogenic acid), iridoids (allobetonicoside and 8-OAc-harpagide), and alkaloids (trigonelline) were identified, characteristic of plants belonging to the genus Stachys. High antioxidant and radical scavenging activities were observed in both in situ and ex vitro acclimated S. scardica plants, correlating with the reported high concentrations of total phenols and flavonoids in these variants. Ex vitro adapted plants also exhibited a well-defined anti-inflammatory potential, demonstrating high inhibitory activity against the complement system. Employing a disk diffusion method, a 100% inhibition effect was achieved compared to positive antibiotic controls against Staphylococcus epidermidis and Propionibacterium acnes, with moderate activity against Bacillus cereus. The induced in vitro and ex vitro model systems can enable the conservation of S. scardica in nature and offer future opportunities for the targeted biosynthesis of valuable secondary metabolites, with potential applications in the pharmaceutical and cosmetic industries. Full article
(This article belongs to the Special Issue Plant Biotechnology Applications in Secondary Metabolite Production)
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17 pages, 3372 KiB  
Article
Untargeted Metabolomics Analysis of the Orchid Species Oncidium sotoanum Reveals the Presence of Rare Bioactive C-Diglycosylated Chrysin Derivatives
by Gianluca Zorzi, Sofia Gambini, Stefano Negri, Flavia Guzzo and Mauro Commisso
Plants 2023, 12(3), 655; https://doi.org/10.3390/plants12030655 - 02 Feb 2023
Cited by 1 | Viewed by 1613
Abstract
Plants are valuable sources of secondary metabolites with pharmaceutical properties, but only a small proportion of plant life has been actively exploited for medicinal purposes to date. Underexplored plant species are therefore likely to contain novel bioactive compounds. In this study, we investigated [...] Read more.
Plants are valuable sources of secondary metabolites with pharmaceutical properties, but only a small proportion of plant life has been actively exploited for medicinal purposes to date. Underexplored plant species are therefore likely to contain novel bioactive compounds. In this study, we investigated the content of secondary metabolites in the flowers, leaves and pseudobulbs of the orchid Oncidium sotoanum using an untargeted metabolomics approach. We observed the strong accumulation of C-diglycosylated chrysin derivatives, which are rarely found in nature. Further characterization revealed evidence of antioxidant activity (FRAP and DPPH assays) and potential activity against neurodegenerative disorders (MAO-B inhibition assay) depending on the specific molecular structure of the metabolites. Natural product bioprospecting in underexplored plant species based on untargeted metabolomics can therefore help to identify novel chemical structures with diverse pharmaceutical properties. Full article
(This article belongs to the Special Issue Plant Biotechnology Applications in Secondary Metabolite Production)
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17 pages, 4299 KiB  
Article
Elicitation of Stilbenes and Benzofuran Derivatives in Hairy Root Cultures of White Mulberry (Morus alba)
by Lingling Fang, Amit Raj Sharma, Chineche Aniemena, Krystian Roedel, Florence Henry, Philippe Moussou, Anita Samuga and Fabricio Medina-Bolivar
Plants 2023, 12(1), 175; https://doi.org/10.3390/plants12010175 - 31 Dec 2022
Cited by 3 | Viewed by 1811
Abstract
Stilbene and benzofuran derivatives isolated from the root of white mulberry (Morus alba) have shown various biological activities, including anti-inflammatory, antioxidant, and antimicrobial properties. The objectives of this study were to develop hairy root cultures and assess the effect of multiple [...] Read more.
Stilbene and benzofuran derivatives isolated from the root of white mulberry (Morus alba) have shown various biological activities, including anti-inflammatory, antioxidant, and antimicrobial properties. The objectives of this study were to develop hairy root cultures and assess the effect of multiple elicitors combinations including (I) methyl-β-cyclodextrin (CD), MgCl2, methyl jasmonate (MeJA), and H2O2, (II) CD, MgCl2, and MeJA and (III) CD, MgCl2, and H2O2, on the production of these bioactive compounds. The highest yields of stilbenes and benzofurans were obtained upon co-treatment with 18 g/L CD, 3 mM H2O2 and 1 mM MgCl2. The stilbenes oxyresveratrol, resveratrol, and 3′-prenylresveratrol accumulated up to 6.27, 0.61, and 5.00 mg/g DW root, respectively. Meanwhile, the aryl benzofurans moracin M and moracin C accumulated up to 7.82 and 1.82 mg/g DW root, respectively. These stilbenes and benzofurans accumulated in the culture medium of the elicited hairy root cultures. They were not detected in the root tissue. However, the oxyresveratrol diglucoside mulberroside A was only detected in the root tissue with yields up to 10.01 mg/g DW. The results demonstrated that co-treatment of white mulberry hairy root cultures with multiple elicitors can significantly enhance production and secretion of stilbenes and benzofurans in this controlled and sustainable axenic culture system. Full article
(This article belongs to the Special Issue Plant Biotechnology Applications in Secondary Metabolite Production)
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14 pages, 1221 KiB  
Article
In Vitro Sprouted Plantlets of Citrullus colocynthis (L.) Schrad Shown to Possess Interesting Levels of Cucurbitacins and Other Bioactives against Pathogenic Fungi
by Belsem Marzouk, Meher Refifà, Serena Montalbano, Annamaria Buschini, Stefano Negri, Mauro Commisso and Francesca Degola
Plants 2022, 11(20), 2711; https://doi.org/10.3390/plants11202711 - 14 Oct 2022
Cited by 2 | Viewed by 1384
Abstract
Cucurbitacins, structurally different triterpenes mainly found in the members of Cucurbitaceae, possess a vast pharmacological potential. Genus Cucurbita, Cucumis, and Citrullus are affluent in these bioactive compounds, and, amongst them, Citrullus colocynthis (L.) Schrad. is widely exploited in folk medicine, since a [...] Read more.
Cucurbitacins, structurally different triterpenes mainly found in the members of Cucurbitaceae, possess a vast pharmacological potential. Genus Cucurbita, Cucumis, and Citrullus are affluent in these bioactive compounds, and, amongst them, Citrullus colocynthis (L.) Schrad. is widely exploited in folk medicine, since a huge number of diseases are successfully treated with organic and aqueous extracts obtained from different organs and tissues of the plant. The well-known pharmacological activities of such species have been attributed to its peculiar composition, which includes cucurbitacins and other bioactive molecules; thus, owing to its high importance as a valuable natural resource for pharmaceuticals and nutraceuticals, C. colocynthis propagation and multiplication protocols are considered significant, but the exploitation of its phytochemical potential is limited by the restricted cultivation conditions and the low rate of seed germination in the natural environment; in fact, the assessment of accumulation rate of specific phytochemicals under controlled conditions is still missing. Axenically sprouted plantlets obtained without the use of culture media or the addition of hormones have been evaluated here for the production of bioactive compounds and relevant bioactive features. Our results proved that derived organic extracts contain cucurbitacins and other bioactives, show antioxidant potential, and exert activity against some pathogenic fungi (Candida krusei, C. albicans, C. parapsilosis, C. glabrata, and Aspergillus flavus), supporting the feasibility of a methodology intended to scale-up cultivation of this species as a source of pharmaceutically interesting compounds, achievable from plantlets cultivated under laboratory conditions. Full article
(This article belongs to the Special Issue Plant Biotechnology Applications in Secondary Metabolite Production)
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17 pages, 796 KiB  
Article
Biotechnological Potential of Different Organs of Mistletoe (Viscum album L.) Collected from Various Host Tree Species in an Urban Area
by Liubov Skrypnik, Pavel Feduraev, Anton Golovin, Pavel Maslennikov, Nikolay Belov, Matvei Matveev and Artem Pungin
Plants 2022, 11(20), 2686; https://doi.org/10.3390/plants11202686 - 12 Oct 2022
Cited by 9 | Viewed by 1898
Abstract
From an economic and ecological standpoint, it is crucial to investigate the biologically active compounds of mistletoe plants, which are currently discarded by pruning urban mistletoe-infested trees. In the present study, the content of phenolic compounds, triterpenic and organic acids, as well as [...] Read more.
From an economic and ecological standpoint, it is crucial to investigate the biologically active compounds of mistletoe plants, which are currently discarded by pruning urban mistletoe-infested trees. In the present study, the content of phenolic compounds, triterpenic and organic acids, as well as the antioxidant activity of the extracts of various mistletoe organs (leaves, stems, and fruits) collected from the most infested tree species were investigated. The mistletoe samples collected from Betula pendula, Acer platanoides, Crataegus monogyna, and Sorbus aucuparia showed the highest content of phenolic acids and flavonoids as well as antioxidant activity, as measured by 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS), and ferric reducing/antioxidant power (FRAP) assays. The leaves and stems of mistletoe from Tilia cordata were characterized by a high content of triterpenic acids (oleanolic, ursolic, and betulinic). The leaves and fruits of mistletoe plants from Populus nigra and Salix alba contained a high concentration of organic acids, particularly succinic and citric acids. Compared to stem and leaf extracts, the antioxidant activity of the mistletoe fruit extracts was 1.5–3 times higher. The obtained results indicate that mistletoe is a valuable raw material and can be used as a source of phenolic compounds and triterpenic and organic acids, as well as for producing extracts with antioxidant properties. Full article
(This article belongs to the Special Issue Plant Biotechnology Applications in Secondary Metabolite Production)
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10 pages, 2175 KiB  
Article
A Phytocomplex Obtained from Salvia officinalis by Cell Culture Technology Effectively Controls the Grapevine Downy Mildew Pathogen Plasmopara viticola
by Isabella Busato, Oriana Bertaiola, Silvio Tundo, Chiara Guarnerio, Marco Lucchetta, Luca Sella, Giovanna Pressi and Francesco Favaron
Plants 2022, 11(20), 2675; https://doi.org/10.3390/plants11202675 - 11 Oct 2022
Cited by 1 | Viewed by 1333
Abstract
The negative impact of using conventional fungicides in plant disease protection has increased the interest in safer alternatives such as plant secondary metabolites, generally having a better toxicological profile. However, cultivation conditions and plant material strongly affect the quality and quantity of secondary [...] Read more.
The negative impact of using conventional fungicides in plant disease protection has increased the interest in safer alternatives such as plant secondary metabolites, generally having a better toxicological profile. However, cultivation conditions and plant material strongly affect the quality and quantity of secondary metabolites obtained from field grown plants, limiting the standardization needed for industrial production. Plant cell culture technology can provide highly homogeneous biomasses with specific chemical characteristics. A phytocomplex with high rosmarinic acid content (10.12% w/w) was obtained from a selected cell line of Salvia officinalis and was tested against the grapevine downy mildew pathogen, Plasmopara viticola. Grapevine leaf discs were sprayed with the phytocomplex at 5 g/L and then inoculated with P. viticola sporangia. Sporulation level on each disc was assessed after 7 days with an image processing software. The phytocomplex reduced by 95% the sporulation level compared to the control and was also more effective than rosmarinic acid alone, used at the same concentration found in the phytocomplex. Persistence of the phytocomplex was also assessed: when applied 5 days before inoculation, it reduced by 90% the sporulation level compared to the control. These results highlight the possibility to take advantage of cell culture techniques to produce safer pesticides with high quality standards. Full article
(This article belongs to the Special Issue Plant Biotechnology Applications in Secondary Metabolite Production)
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12 pages, 2898 KiB  
Article
Silencing of Dihydroflavonol 4-reductase in Chrysanthemum Ray Florets Enhances Flavonoid Biosynthesis and Antioxidant Capacity
by Sun-Hyung Lim, Da-Hye Kim, Jae-A Jung, Nam-In Hyung, YeoJin Youn and Jong-Yeol Lee
Plants 2022, 11(13), 1681; https://doi.org/10.3390/plants11131681 - 24 Jun 2022
Cited by 6 | Viewed by 1512
Abstract
Flavonoid biosynthesis requires the activities of several enzymes, which form weakly-bound, ordered protein complexes termed metabolons. To decipher flux regulation in the flavonoid biosynthetic pathway of chrysanthemum (Chrysanthemum morifolium Ramat), we suppressed the gene-encoding dihydroflavonol 4-reductase (DFR) through RNA interference (RNAi)-mediated post-transcriptional [...] Read more.
Flavonoid biosynthesis requires the activities of several enzymes, which form weakly-bound, ordered protein complexes termed metabolons. To decipher flux regulation in the flavonoid biosynthetic pathway of chrysanthemum (Chrysanthemum morifolium Ramat), we suppressed the gene-encoding dihydroflavonol 4-reductase (DFR) through RNA interference (RNAi)-mediated post-transcriptional gene silencing under a floral-specific promoter. Transgenic CmDFR-RNAi chrysanthemum plants were obtained by Agrobacterium-mediated transformation. Genomic PCR analysis of CmDFR-RNAi chrysanthemums propagated by several rounds of stem cuttings verified stable transgene integration into the genome. CmDFR mRNA levels were reduced by 60–80% in CmDFR-RNAi lines compared to those in wild-type (WT) plants in ray florets, but not leaves. Additionally, transcript levels of flavonoid biosynthetic genes were highly upregulated in ray florets of CmDFR-RNAi chrysanthemum relative to those in WT plants, while transcript levels in leaves were similar to WT. Total flavonoid contents were high in ray florets of CmDFR-RNAi chrysanthemums, but flavonoid contents of leaves were similar to WT, consistent with transcript levels of flavonoid biosynthetic genes. Ray florets of CmDFR-RNAi chrysanthemums exhibited stronger antioxidant capacity than those of WT plants. We propose that post-transcriptional silencing of CmDFR in ray florets modifies metabolic flux, resulting in enhanced flavonoid content and antioxidant activity. Full article
(This article belongs to the Special Issue Plant Biotechnology Applications in Secondary Metabolite Production)
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17 pages, 1895 KiB  
Article
Inhibition of Human Monoamine Oxidases A and B by Specialized Metabolites Present in Fresh Common Fruits and Vegetables
by Claudio Marcello Marzo, Sofia Gambini, Stefania Poletti, Francesca Munari, Michael Assfalg and Flavia Guzzo
Plants 2022, 11(3), 346; https://doi.org/10.3390/plants11030346 - 27 Jan 2022
Cited by 5 | Viewed by 2993
Abstract
Diets rich in fruits and vegetables are associated with better psychological wellbeing and cognitive functions, although it is unclear which molecules and mechanisms are involved. One potential explanation is the inhibition of monoamine oxidases (MAOs), which have been linked to several neurological disorders. [...] Read more.
Diets rich in fruits and vegetables are associated with better psychological wellbeing and cognitive functions, although it is unclear which molecules and mechanisms are involved. One potential explanation is the inhibition of monoamine oxidases (MAOs), which have been linked to several neurological disorders. The present study investigated the ability of kiwifruit to inhibit MAO-A and MAO-B, refining an in vitro assay to avoid confounding effects. Ultra-performance liquid chromatography/mass spectrometry (UPLC-QTOF) and nuclear magnetic resonance spectroscopy (NMR) were used to select individual kiwifruit metabolites for further analysis. Moreover, extracts of other common fruits and vegetables were screened to identify promising candidate inhibitors. Multiple extracts and compounds inhibited both enzymes, and the selective inhibition of MAO-B by the major kiwifruit specialized metabolite D-(−)-quinic acid was observed. These results suggest that fruits and vegetables contain metabolites that inhibit the activity of MAO-A and -B, offering a potential natural option for the treatment of neurological disorders, in which MAOs are involved. Full article
(This article belongs to the Special Issue Plant Biotechnology Applications in Secondary Metabolite Production)
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14 pages, 3056 KiB  
Article
Undifferentiated In Vitro Cultured Actinidia deliciosa as Cell Factory for the Production of Quercetin Glycosides
by Stefano Negri, Sofia Gambini, Stefania Ceoldo, Linda Avesani, Mauro Commisso and Flavia Guzzo
Plants 2021, 10(11), 2499; https://doi.org/10.3390/plants10112499 - 18 Nov 2021
Cited by 4 | Viewed by 2055
Abstract
Land plants produce a vast arsenal of specialized metabolites and many of them display interesting bioactivities in humans. Recently, flavonol quercetin gained great attention in the light of the COVID-19 pandemic because, in addition to the anti-inflammatory, antiviral and anti-cancer activity already described, [...] Read more.
Land plants produce a vast arsenal of specialized metabolites and many of them display interesting bioactivities in humans. Recently, flavonol quercetin gained great attention in the light of the COVID-19 pandemic because, in addition to the anti-inflammatory, antiviral and anti-cancer activity already described, it emerged as possible inhibitor of 3CLpro, the major protease of SARS-CoV-2 virus. Plant cell and tissue culture (PCTC) is an attractive platform for the biotechnological production of plant metabolites. This technology allows a large amount of water and agricultural land to be saved and, being free of contaminants in the process, it is suitable for scaling up the production in bioreactors. In a project aimed to generate and screen in vitro plant cells for the production of valuable specialized metabolites for commercial production, we generated various cell lines from Actinidia deliciosa (kiwi fruit tree) and Actinidia chinensis (gold kiwi fruit tree), that were able to produce relevant amounts of quercetin derivatives, mainly quercetin glycosides. Three cell lines from A. deliciosa were characterized by targeted and untargeted metabolomics. In standard growing conditions, they produce and accumulate up to 13.26 mg/100 g fresh weight (419.76 mg/100 g dry weight) of quercetin derivatives. To address future industrial applications, these cell lines should be entered into an acceleration program to further increase the amount of these metabolites by optimizing the culture conditions and elicitation. Full article
(This article belongs to the Special Issue Plant Biotechnology Applications in Secondary Metabolite Production)
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23 pages, 1410 KiB  
Article
The Combination of Untargeted Metabolomics and Machine Learning Predicts the Biosynthesis of Phenolic Compounds in Bryophyllum Medicinal Plants (Genus Kalanchoe)
by Pascual García-Pérez, Leilei Zhang, Begoña Miras-Moreno, Eva Lozano-Milo, Mariana Landin, Luigi Lucini and Pedro P. Gallego
Plants 2021, 10(11), 2430; https://doi.org/10.3390/plants10112430 - 10 Nov 2021
Cited by 10 | Viewed by 3008
Abstract
Phenolic compounds constitute an important family of natural bioactive compounds responsible for the medicinal properties attributed to Bryophyllum plants (genus Kalanchoe, Crassulaceae), but their production by these medicinal plants has not been characterized to date. In this work, a combinatorial approach including [...] Read more.
Phenolic compounds constitute an important family of natural bioactive compounds responsible for the medicinal properties attributed to Bryophyllum plants (genus Kalanchoe, Crassulaceae), but their production by these medicinal plants has not been characterized to date. In this work, a combinatorial approach including plant tissue culture, untargeted metabolomics, and machine learning is proposed to unravel the critical factors behind the biosynthesis of phenolic compounds in these species. The untargeted metabolomics revealed 485 annotated compounds that were produced by three Bryophyllum species cultured in vitro in a genotype and organ-dependent manner. Neurofuzzy logic (NFL) predictive models assessed the significant influence of genotypes and organs and identified the key nutrients from culture media formulations involved in phenolic compound biosynthesis. Sulfate played a critical role in tyrosol and lignan biosynthesis, copper in phenolic acid biosynthesis, calcium in stilbene biosynthesis, and magnesium in flavanol biosynthesis. Flavonol and anthocyanin biosynthesis was not significantly affected by mineral components. As a result, a predictive biosynthetic model for all the Bryophyllum genotypes was proposed. The combination of untargeted metabolomics with machine learning provided a robust approach to achieve the phytochemical characterization of the previously unexplored species belonging to the Bryophyllum subgenus, facilitating their biotechnological exploitation as a promising source of bioactive compounds. Full article
(This article belongs to the Special Issue Plant Biotechnology Applications in Secondary Metabolite Production)
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18 pages, 3185 KiB  
Article
Shape Matters: Plant Architecture Affects Chemical Uniformity in Large-Size Medical Cannabis Plants
by Nadav Danziger and Nirit Bernstein
Plants 2021, 10(9), 1834; https://doi.org/10.3390/plants10091834 - 03 Sep 2021
Cited by 21 | Viewed by 16133
Abstract
Since plant organs sense their environment locally, gradients of micro-climates in the plant shoot may induce spatial variability in the physiological state of the plant tissue and hence secondary metabolism. Therefore, plant architecture, which affects micro-climate in the shoot, may considerably affect the [...] Read more.
Since plant organs sense their environment locally, gradients of micro-climates in the plant shoot may induce spatial variability in the physiological state of the plant tissue and hence secondary metabolism. Therefore, plant architecture, which affects micro-climate in the shoot, may considerably affect the uniformity of cannabinoids in the Cannabis sativa plant, which has significant pharmaceutical and economic importance. Variability of micro-climates in plant shoots intensifies with the increase in plant size, largely due to an increase in inter-shoot shading. In this study, we therefore focused on the interplay between shoot architecture and the cannabinoid profile in large cannabis plants, ~2.5 m in height, with the goal to harness architecture modulation for the standardization of cannabinoid concentrations in large plants. We hypothesized that (i) a gradient of light intensity along the plants is accompanied by changes to the cannabinoid profile, and (ii) manipulations of plant architecture that increase light penetration to the plant increase cannabinoid uniformity and yield biomass. To test these hypotheses, we investigated effects of eight plant architecture manipulation treatments involving branch removals, defoliation, and pruning on plant morpho-physiology, inflorescence yield, cannabinoid profile, and uniformity. The results revealed that low cannabinoid concentrations in inflorescences at the bottom of the plants correlate with low light penetration, and that increasing light penetration by defoliation or removal of bottom branches and leaves increases cannabinoid concentrations locally and thereby through spatial uniformity, thus supporting the hypotheses. Taken together, the results reveal that shoot architectural modulation can be utilized to increase cannabinoid standardization in large cannabis plants, and that the cannabinoid profile in an inflorescence is an outcome of exogenous and endogenous factors. Full article
(This article belongs to the Special Issue Plant Biotechnology Applications in Secondary Metabolite Production)
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10 pages, 737 KiB  
Communication
PEG 6000-Stimulated Drought Stress Improves the Attributes of In Vitro Growth, Steviol Glycosides Production, and Antioxidant Activities in Stevia rebaudiana Bertoni
by Muhammad Arslan Ahmad, Rabia Javed, Muhammad Adeel, Muhammad Rizwan and Yuesuo Yang
Plants 2020, 9(11), 1552; https://doi.org/10.3390/plants9111552 - 12 Nov 2020
Cited by 59 | Viewed by 5763
Abstract
This study addresses the favourable effects of drought stress imposed by polyethylene glycol (PEG) 6000 on the micropropagated shoots of Stevia rebaudiana. Various concentrations, i.e., 0, 0.5, 1, 2, and 4% of PEG 6000 were applied to the nodal shoot explants for [...] Read more.
This study addresses the favourable effects of drought stress imposed by polyethylene glycol (PEG) 6000 on the micropropagated shoots of Stevia rebaudiana. Various concentrations, i.e., 0, 0.5, 1, 2, and 4% of PEG 6000 were applied to the nodal shoot explants for four weeks, and the influence produced on shoots growth parameters, bioactive steviol glycosides (rebaudioside A and stevioside), and nonenzymatic antioxidant activities (total phenolic content (TPC), total flavonoid content (TFC), total antioxidant capacity (TAC), total reducing power (TRP) and 1,1-diphenyl-2-picrylhydrazyl(DPPH)-free radical scavenging activity (FRSA)) was elucidated. The significantly highest yield (92.4% direct shoot organogenesis) and secondary metabolites (2.94% Reb A, 2.52% ST, 95.3% DPPH-FRSA, 15.0% TPC, 13.0 µg/mg TFC, 22.3 µg/mg TAC, and 19.8 µg/mg TRP) production in response to abiotic stress elicitors was obtained in Murashige and Skoog (MS) medium treatment provided with 4% of PEG 6000. The overall trend was significant enhancement of growth dynamics and pharmaceutical compounds from control to 4% of PEG 6000 concentration as a defensive response against reactive oxygen species (ROS) produced in excess by water deficit. This is a very promising study to be employed in bioreactors to get markedly enhanced content of compounds of medicinal importance in the pharmaceutical market. Full article
(This article belongs to the Special Issue Plant Biotechnology Applications in Secondary Metabolite Production)
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10 pages, 3142 KiB  
Article
Analysis of the Antimicrobial, Cytotoxic, and Antioxidant Activities of Cnidium officinale Extracts
by Myung-Jin Lee and Min-Kyung Kang
Plants 2020, 9(8), 988; https://doi.org/10.3390/plants9080988 - 04 Aug 2020
Cited by 8 | Viewed by 2806
Abstract
This study analyzed the antimicrobial, cytotoxic, and antioxidant properties of Cnidiumofficinale (CO) extracts to confirm their antimicrobial activity toward oral microorganisms. The control group contained 0 μg/mL of CO, and the experimental groups contained 50, 100, 150, and 200 μg/mL of CO. [...] Read more.
This study analyzed the antimicrobial, cytotoxic, and antioxidant properties of Cnidiumofficinale (CO) extracts to confirm their antimicrobial activity toward oral microorganisms. The control group contained 0 μg/mL of CO, and the experimental groups contained 50, 100, 150, and 200 μg/mL of CO. To confirm the antibacterial activity of CO extracts against microorganisms in the oral cavity, an inhibition zone test, a colony-forming unit (CFU) analysis, an optical density (OD) evaluation, and a SEM (scanning electron microscopy) analysis were performed. A cytotoxicity test was also conducted to determine cell viability, and the contents of flavonoids and polyphenols were measured to analyze the extract components. In the control group, the growth inhibition zone increased, while the CFU and OD values decreased (p < 0.05). The SEM analysis confirmed that the number of microorganisms for both the microbes decreased. The cell viability was more than 80% in both the control and experimental groups, excluding the 200 μg/mL sample. The flavonoid and polyphenol contents in the experimental groups showed higher values than those of the control group. Therefore, the CO extract showed considerable antimicrobial activity toward both Streptococcus mutans and Candida albicans, suggesting that it may be used as a natural antimicrobial agent for dental applications. Full article
(This article belongs to the Special Issue Plant Biotechnology Applications in Secondary Metabolite Production)
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11 pages, 1871 KiB  
Article
Ecdysteroid Content and Therapeutic Activity in Elicited Spinach Accessions
by Jonathan Gorelick, Rona Hacohen Iraqi and Nirit Bernstein
Plants 2020, 9(6), 727; https://doi.org/10.3390/plants9060727 - 09 Jun 2020
Cited by 10 | Viewed by 3089
Abstract
While spinach is an established nutritionally important crop, its medicinal value is not as well known. Spinach is rich in ecdysteroids, insect hormone analogs with a number of medicinal properties including anti-oxidative, anti-inflammatory and even anabolic activity. However, the potential of spinach as [...] Read more.
While spinach is an established nutritionally important crop, its medicinal value is not as well known. Spinach is rich in ecdysteroids, insect hormone analogs with a number of medicinal properties including anti-oxidative, anti-inflammatory and even anabolic activity. However, the potential of spinach as a medicinal plant has not yet been developed. In this study, the ecdysteroid content of spinach was optimized to increase its therapeutic value. Spinach seeds from various sources were grown under controlled hydroponic conditions and analyzed for ecdysteroid content and related anabolic activity. Variations in ecdysteroid content and the related anabolic activity were observed among spinach accessions. A selected variety, Spinacia oleracea cv. Turkey, was exposed to various physical and chemical elicitors to increase and stabilize ecdysteroid content. A number of elicitors, including methyl salicylate and mechanical damage, significantly increased ecdysteroid content and anabolic activity 24 h after exposure. The effect was transient and disappeared 48 h thereafter. Further work is needed to identify the most suitable germplasm and elicitation conditions for optimal ecdysteroid content. Full article
(This article belongs to the Special Issue Plant Biotechnology Applications in Secondary Metabolite Production)
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12 pages, 1933 KiB  
Article
Enhanced Production of Bryonolic Acid in Trichosanthes cucumerina L. (Thai Cultivar) Cell Cultures by Elicitors and Their Biological Activities
by Pornpatsorn Lertphadungkit, Jiraphong Suksiriworapong, Veena Satitpatipan, Supaart Sirikantaramas, Amaraporn Wongrakpanich and Somnuk Bunsupa
Plants 2020, 9(6), 709; https://doi.org/10.3390/plants9060709 - 02 Jun 2020
Cited by 11 | Viewed by 3672
Abstract
Bryonolic acid is a triterpenoid compound found in cucurbitaceous roots. Due to its biological activities, this compound gets more attention to improve production. Herein, we carried out efficient ways with high bryonolic acid productions from Trichosanthes cucumerina L., a Thai medicinal plant utilizing [...] Read more.
Bryonolic acid is a triterpenoid compound found in cucurbitaceous roots. Due to its biological activities, this compound gets more attention to improve production. Herein, we carried out efficient ways with high bryonolic acid productions from Trichosanthes cucumerina L., a Thai medicinal plant utilizing plant cell cultures. The results showed that calli (24.65 ± 1.97 mg/g dry weight) and cell suspensions (15.69 ± 0.78 mg/g dry weight) exhibited the highest bryonolic acid productions compared with natural roots (approximately 2 mg/g dry weight). In the presence of three elicitors (methyl jasmonate, yeast extract, and chitosan), cell suspensions treated with 1 mg/mL of chitosan for eight days led to higher bryonolic acid contents (23.56 ± 1.68 mg/g dry weight). Interestingly, cell culture and root extracts with high bryonolic acid contents resulted in significantly higher percent cell viabilities than those observed under control (1% v/v DMSO) treatment in Saos-2 and MCF-7 cells. The present study indicated that T. cucumerina L. cell cultures are alternative and efficient to produce the biologically important secondary metabolite. Full article
(This article belongs to the Special Issue Plant Biotechnology Applications in Secondary Metabolite Production)
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15 pages, 442 KiB  
Article
Phytochemistry, Toxicology and Therapeutic Value of Petasites hybridus Subsp. Ochroleucus (Common Butterbur) from the Balkans
by Tatjana Mihajilov-Krstev, Boris Jovanović, Bojan Zlatković, Jelena Matejić, Jelena Vitorović, Vladimir Cvetković, Budimir Ilić, Ljubiša Đorđević, Nataša Joković, Dragoljub Miladinović, Tatjana Jakšić, Nemanja Stanković, Vesna Stankov Jovanović and Nirit Bernstein
Plants 2020, 9(6), 700; https://doi.org/10.3390/plants9060700 - 31 May 2020
Cited by 12 | Viewed by 3433
Abstract
Petasites hybridus (Common butterbur) is extensively used in traditional medicine, and is currently gaining interest and popularity as a food supplement and for its medicinal properties. It contains a large number of active compounds of potential therapeutic activity, but also toxic pyrrolizidine alkaloids. [...] Read more.
Petasites hybridus (Common butterbur) is extensively used in traditional medicine, and is currently gaining interest and popularity as a food supplement and for its medicinal properties. It contains a large number of active compounds of potential therapeutic activity, but also toxic pyrrolizidine alkaloids. Science-based information is needed to support the developing modern use of this plant, and to direct continued safe practice in traditional medicine. The present study focused on the essential oils from leaves and rhizomes of the understudied P. hybridus ssp. ochroleucus from the Balkans, and evaluated its phytochemistry and potential therapeutic activities (antimicrobial, antioxidant, anti-cholinesterase and anti-inflammatory), as well its toxicology potential (acute toxicity in insects and mice). We studied the essential oils, which are not commonly used in traditional practices, but have a potential for safe use since the toxic pyrrolizidine alkaloids, which are non-volatiles, are usually not present in the distilled essential oils. Pyrrolizidine alkaloids were indeed not detected in the essential oils; ingestion of the essential oils did not induce toxicity signs in mice, and topical application did not elicit skin irritation in humans. The essential oils had no antimicrobial properties against 20 pathogenic bacterial strains, but demonstrated good local anti-inflammatory activity in a Carrageenan-induced paw edema test. An insect toxicity test demonstrated that the leaf essential oil is an efficient insect repellent, and the demonstrated anti-cholinesterase activity suggests a potential for the treatment of neurological conditions. Isopetasin, a sesquiterpene found in plants of the genus Petasites, known to have anti-inflammatory effects, was present only in the rhizomes essential oil (3.9%), and sesquiterpene lactones concentrations were high, likely contributing to the antioxidant activity. Full article
(This article belongs to the Special Issue Plant Biotechnology Applications in Secondary Metabolite Production)
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Review

Jump to: Research

25 pages, 1304 KiB  
Review
The Integral Boosting Effect of Selenium on the Secondary Metabolism of Higher Plants
by Liubov Skrypnik, Pavel Feduraev, Anton Golovin, Pavel Maslennikov, Tatiana Styran, Maria Antipina, Anastasiia Riabova and Dmitriy Katserov
Plants 2022, 11(24), 3432; https://doi.org/10.3390/plants11243432 - 08 Dec 2022
Cited by 2 | Viewed by 1636
Abstract
Selenium is a micronutrient with a wide range of functions in animals, including humans, and in microorganisms such as microalgae. However, its role in plant metabolism remains ambiguous. Recent studies of Se supplementation showed that not only does it increase the content of [...] Read more.
Selenium is a micronutrient with a wide range of functions in animals, including humans, and in microorganisms such as microalgae. However, its role in plant metabolism remains ambiguous. Recent studies of Se supplementation showed that not only does it increase the content of the element itself, but also affects the accumulation of secondary metabolites in plants. The purpose of this review is to analyze and summarize the available data on the place of selenium in the secondary metabolism of plants and its effect on the accumulation of some plant metabolites (S- and N-containing secondary metabolites, terpenes, and phenolic compounds). In addition, possible molecular mechanisms and metabolic pathways underlying these effects are discussed. It should be noted that available data on the effect of Se on the accumulation of secondary metabolites are inconsistent and contradictory. According to some studies, selenium has a positive effect on the accumulation of certain metabolites, while other similar studies show a negative effect or no effect at all. The following aspects were identified as possible ways of regulating plant secondary metabolism by Se-supplementation: changes occurring in primary S/N metabolism, hormonal regulation, redox metabolism, as well as at the transcriptomic level of secondary metabolite biosynthesis. In all likelihood, the confusion in the results can be explained by other, more complex regulatory mechanisms in which selenium is involved and which affect the production of metabolites. Further study on the involvement of various forms of selenium in metabolic and signaling pathways is crucial for a deeper understanding of its role in growth, development, and health of plants, as well as the regulatory mechanisms behind them. Full article
(This article belongs to the Special Issue Plant Biotechnology Applications in Secondary Metabolite Production)
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38 pages, 2270 KiB  
Review
Root Cultures, a Boon for the Production of Valuable Compounds: A Comparative Review
by Masooma Jawad Hussain, Yawar Abbas, Naushaba Nazli, Sara Fatima, Samantha Drouet, Christophe Hano and Bilal Haider Abbasi
Plants 2022, 11(3), 439; https://doi.org/10.3390/plants11030439 - 05 Feb 2022
Cited by 27 | Viewed by 4289
Abstract
Medicinal plants are an inevitable source of pharmaceutical drugs and most of the world population depends on these plants for health benefits. The increasing global demand for bioactive compounds from medicinal plants has posed a great threat to their existence due to overexploitation. [...] Read more.
Medicinal plants are an inevitable source of pharmaceutical drugs and most of the world population depends on these plants for health benefits. The increasing global demand for bioactive compounds from medicinal plants has posed a great threat to their existence due to overexploitation. Adventitious root and hairy root culture systems are an alternative approach to the conventional method for mass production of valuable compounds from medicinal plants owing to their rapid growth, biosynthetic and genetic stability. The main purpose of this review is to investigate the recent scientific research published worldwide on the application of adventitious and hairy root cultures to produce valuable compounds from medicinal plants. Furthermore, a comparison of adventitious root vs. hairy root cultures to produce valuable compounds has also been discussed. Various aspects such as medium composition, carbon source, pH, amount of macronutrients, optimization strategy, scale-up cultures, and use of biotic abiotic and nano-elicitors at various concentrations are the topic of discussion in this review. Several studies on adventitious and hairy root cultures of Polygonum multiflorum¸ Withania somnifera¸ Echinacea purpurea and Ajuga bracteosa have been discussed in detail which highlights the importance of elicitation strategies and bioreactor system, presenting commercial applications. Full article
(This article belongs to the Special Issue Plant Biotechnology Applications in Secondary Metabolite Production)
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