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Production of Secondary Metabolites In Vitro

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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 March 2023) | Viewed by 16657

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Special Issue Editors

Dr. Katerina Grigoriadou

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Guest Editor

Laboratory of Protection and Evaluation of Native and Floricultural Species, Balkan Botanic Garden of Kroussia, Institute of Plant Breeding and Genetic Resources, Hellenic Agricultural Organization (HAO)-DEMETER, P.C. 570 01 Thermi, P.O. Box 60458 Thessaloniki, Greece
Interests: plant propagation; in vitro culture; micropropagation; production of secondary metabolites in vitro; medicinal plants; ex situ conservation; native flora
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Dr. Eirini Sarrou

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Co-Guest Editor

Institute of Plant Breeding and Genetic Resources, Hellenic Agricultural Organization (HAO)-DEMETER, P.C. 570 01 Thermi, P.O. Box 60458 Thessaloniki, Greece
Interests: medicinal and aromatic plant breeding; plants secondary metabolites; metabolomics; functional genomics

Dr. Eleni Maloupa

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Co-Guest Editor

Laboratory of Protection and Evaluation of Native and Floricultural Species, Balkan Botanic Garden of Kroussia, Institute of Plant Breeding and Genetic Resources, Hellenic Agricultural Organization (HAO)-DEMETER, P.C. 570 01 Thermi, P.O. Box 60458 Thessaloniki, Greece
Interests: plant propagation; plant sustainable exploitation; Greek aromatic-medicinal crops; Greek plants with ornamental value; Greek plants with nutritional value; ex-situ plant conservation; soilless culture

Special Issue Information

Dear Colleagues,

Secondary plant metabolites are not essential to plant growth, but are required for the plant to survive in its environment. They are produced in small amounts, usually have very complicated structures and often accumulate in specialized tissues. Secondary metabolites are used as especially chemicals such as drugs, flavours, fragrances, insecticides, and dyes and thus have a great economic value. Plants, such as medicinal/aromatic plants (MAPs) contain various secondary metabolites (e.g., alkaloids, flavonoids, glycosides, phenols, saponins), in their tissues, commercially valuable. An alternative to agricultural processes for producing secondary metabolites, is the in vitro culture, running in controlled environment, independently from climate and soil conditions. It is still in its infancy the knowledge of biosynthetic pathways of phytochemicals and desired phytochemicals in plants. This Special Issue of Plants will highlight the significance, function, complexity and potential applications of secondary metabolites that are produced in vitro and will suggest strategies to improve the in vitro production of secondary metabolites.

Dr. Katerina Grigoriadou
Dr. Eleni Maloupa
Dr. Eirini Sarrou

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.

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Dr. Katerina Grigoriadou
Dr. Eleni Maloupa
Dr. Eirini Sarrou
Guest Editors

Manuscript Submission Information

Keywords

medicinal/aromatic plants
micropropagation
in vitro plant material
developmental stages
secondary metabolites
bioactive compounds
essential oils
volatiles
isolation of metabolites, metabolomics
extraction of metabolites
culture medium
secretory activity
secretory rate
elicitors
enzyme activity
metabolic pathways
metabolic engineering
bioreactors

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Published Papers (5 papers)

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Research

16 pages, 3595 KiB

Open AccessEditor’s ChoiceArticle

Antitumor and Antioxidant Activities of In Vitro Cultivated and Wild-Growing Clinopodium vulgare L. Plants

by Maria Petrova, Lyudmila Dimitrova, Margarita Dimitrova, Petko Denev, Desislava Teneva, Ani Georgieva, Polina Petkova-Kirova, Maria Lazarova and Krasimira Tasheva

Plants 2023, 12(8), 1591; https://doi.org/10.3390/plants12081591 - 9 Apr 2023

Cited by 7 | Viewed by 3125

Abstract

Clinopodium vulgare L. is a valuable medicinal plant used for its anti-inflammatory, antibacterial and wound-healing properties. The present study describes an efficient protocol for the micropropagation of C. vulgare and compares, for the first time, the chemical content and composition and antitumor and antioxidant activities of extracts from in vitro cultivated and wild-growing plants. The best nutrient medium was found to be Murashige and Skoog (MS) supplemented with 1 mg/L BAP and 0.1 IBA mg/L, yielding on average 6.9 shoots per nodal segment. Flower aqueous extracts from in vitro plants had higher total polyphenol content (29,927.6 ± 592.1 mg/100 g vs. 27,292.8 ± 85.3 mg/100 g) and ORAC antioxidant activity (7281.3 ± 82.9 µmol TE/g vs. 7246.3 ± 62.4 µmol TE/g) compared to the flowers of wild plants. HPLC detected qualitative and quantitative differences in phenolic constituents between the in vitro cultivated and wild-growing plants’ extracts. Rosmarinic acid was the major phenolic constituent, being accumulated mainly in leaves, while neochlorogenic acid was a major compound in the flowers of cultivated plants. Catechin was found only in cultivated plants, but not in wild plants or cultivated plants’ stems. Aqueous extracts of both cultivated and wild plants showed significant in vitro antitumor activity against human HeLa (cervical adenocarcinoma), HT-29 (colorectal adenocarcinoma) and MCF-7 (breast cancer) cell lines. The best cytotoxic activity against most of the cancer cell lines, combined with the least detrimental effects on a non-tumor human keratinocyte cell line (HaCaT), was shown by the leaf (250 µg/mL) and flower (500 µg/mL) extracts of cultivated plants, making cultivated plants a valuable source of bioactive compounds and a suitable candidate for anticancer therapy. Full article

(This article belongs to the Special Issue Production of Secondary Metabolites In Vitro)

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17 pages, 1940 KiB

Open AccessArticle

Farm or Lab? A Comparative Study of Oregano’s Leaf and Callus Volatile Isolates Chemistry and Cytotoxicity

by Antonis Kakalis, Vasileios Tsekouras, Sofia Mavrikou, Georgia Moschopoulou, Spyridon Kintzios, Epameinondas Evergetis, Vasilios Iliopoulos, Sofia D. Koulocheri and Serkos A. Haroutounian

Plants 2023, 12(7), 1472; https://doi.org/10.3390/plants12071472 - 28 Mar 2023

Cited by 1 | Viewed by 3695

Abstract

Oregano (Origanum vulgare, Lamiaceae plant family) is a well-known aromatic herb with great commercial value, thoroughly utilized by food and pharmaceutical industries. The present work regards the comparative assessment of in vitro propagated and commercially available oregano tissue natural products. This study includes their secondary metabolites’ biosynthesis, antioxidant properties, and anticancer activities. The optimization of callus induction from derived oregano leaf explants and excessive oxidative browning was performed using various plant growth regulators, light conditions, and antioxidant compounds. The determination of oregano callus volatiles against the respective molecules in maternal herbal material was performed using gas chromatography–mass spectrometry (GC/MS) analysis. In total, the presence of twenty-seven phytochemicals was revealed in both leaf and callus extracts, from which thirteen molecules were biosynthesized in both tissues studied, seven compounds were present only in callus extracts, and seven metabolites only in leaf extracts. Carvacrol and sabinene hydrate were the prevailing volatiles in all tissues exploited, along with alkanes octacosane and triacontane and the trimethylsilyl (TMS) derivative of carvacrol that were detected in significant amounts only in callus extracts. The MTT assay was employed to assess the in vitro cytotoxic properties of oregano extracts against the epithelial human breast cancer MDA-MB-231 and the human neuroblastoma SK-N-SH cell lines. The extracts displayed concentration and time-dependent responses in cell proliferation rates. Full article

(This article belongs to the Special Issue Production of Secondary Metabolites In Vitro)

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16 pages, 1270 KiB

Open AccessArticle

Rosmarinic Acid Production from Origanum dictamnus L. Root Liquid Cultures In Vitro

by Virginia Sarropoulou, Charikleia Paloukopoulou, Anastasia Karioti, Eleni Maloupa and Katerina Grigoriadou

Plants 2023, 12(2), 299; https://doi.org/10.3390/plants12020299 - 8 Jan 2023

Cited by 3 | Viewed by 2067

Abstract

In the present work Origanum dictamnus L. was studied as a suitable in vitro adventitious root culture system for the production of important bioactive molecules, such as rosmarinic acid (RA). Callus culture was initiated from leaf, petiole and root explants on solid MS medium supplemented with either 5 μM NAA + 5 μM kinetin (ODK3) or 5 μM NAA + 0.5 μM kinetin (ODK4). New roots formed from leaf, petiole and root calluses were aseptically transferred into Erlenmeyer flasks containing 100 mL liquid medium and shaken at 120 rpm in the dark. The liquid medium used was the MS supplemented either with 35 μM IBA + 2.5 μM kinetin (ODY1) or 5 μM NAA + 0.5 μM kinetin (ODY2). Biomass production parameters, RA content (%) and yield index (YI) were recorded for each treatment explant type, medium composition and incubation period. Results showed, in every case, the production of RA in vitro. Between the two liquid media (ODY1, ODY2) and the different culture periods, the ODY1 medium and the longest 200-day-culture period were more effective for RA and biomass production, regardless of the initial explant type used. The combination of ODK4-ODY1 resulted in higher RA (5.1% and 4.7%), fresh biomass production (19.0 g and 11.6 g), mean YI (93.7 mg and 51.4 mg) and YI per explant (3.75 mg and 2.06 mg) for roots derived from leaf calluses and root calluses, respectively. However, the solid ODK3 (200 days)–liquid ODY1 (40 days) transition treatment was more beneficial for roots derived from petiole calluses leading to an 18.8-fold increase in fresh biomass growth rate. RA accumulation and YIs were also significantly influenced by explant type, with the highest value produced from root petiole calluses (6.6% RA dry weight, 115.3 mg mean YI and 4.61 mg YI per explant) after 240 days. Full article

(This article belongs to the Special Issue Production of Secondary Metabolites In Vitro)

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14 pages, 1298 KiB

Open AccessArticle

Heterologous Expression of Three Transcription Factors Differently Regulated Astragalosides Metabolic Biosynthesis in Astragalus membranaceus Hairy Roots

by Xiao Hua Li, Jae Kwang Kim and Sang Un Park

Plants 2022, 11(14), 1897; https://doi.org/10.3390/plants11141897 - 21 Jul 2022

Cited by 9 | Viewed by 2641

Abstract

Astragalus membranaceus has been used as a highly popular Chinese herbal medicine for centuries. Triterpenoids, namely astragalosides I, II, III, and IV, represent the main active compounds in this plant species. Transcription factors have a powerful effect on metabolite biosynthesis in plants. We investigated the effect of the Arabidopsis MYB12, production of anthocyanin pigment 1 (PAP1), and maize leaf color (LC) transcription factors in regulating the synthesis of astragaloside metabolites in A. membranaceus. Overexpression of these transcription factors in hairy roots differentially up-regulated these active compounds. Specifically, the overexpression of LC resulted in the accumulation of astragalosides I–IV. The content of astragalosides I and IV were, in particular, more highly accumulated. Overexpression of MYB12 increased the accumulation of astragaloside I in transgenic hairy roots, followed by astragaloside IV, and overexpression of PAP1 resulted in the increased synthesis of astragalosides I and IV. In addition, we found that overexpression of PAP1 together with LC increased astragaloside III levels. At the transcriptional level, several key genes of the mevalonate biosynthetic pathway, especially HMGR1, HMGR2, and HMGR3, were up-regulated differentially in response to these transcription factors, resulting in astragaloside synthesis in the hairy roots of A. membranaceus. Overall, our results indicated that heterologous expression of Arabidopsis MYB12, PAP1, and maize LC differentially affected triterpenoids biosynthesis, leading to the increased biosynthesis of active compounds in A. membranaceus. Full article

(This article belongs to the Special Issue Production of Secondary Metabolites In Vitro)

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11 pages, 4208 KiB

Open AccessArticle

Development of Hairy Root Cultures for Biomass and Triterpenoid Production in Centella asiatica

by Seungeun Baek, Jong-Eun Han, Thanh-Tam Ho and So-Young Park

Plants 2022, 11(2), 148; https://doi.org/10.3390/plants11020148 - 6 Jan 2022

Cited by 23 | Viewed by 4065

Abstract

Centella asiatica (Apiaceae) is a tropical/subtropical medicinal plant, which contains a variety of triterpenoids, including madecassoside, asiaticoside, madecassic acid, and asiatic acid. In this study, we tested the efficiency of hairy root (HR) induction in C. asiatica from leaf and petiole explants. Leaves and petioles collected from C. asiatica plants were suspended in agro-stock for 30 min and co-cultured with Agrobacterium rhizogenes for 3 days to induce HR formation. The transformation efficiency of leaf and petiole explants was approximately 27% and 12%, respectively. A total of 36 HR lines were identified by PCR-based amplification of rol genes, and eight of these lines were selected for further analysis. Among all eight HR lines, the petiole-derived lines HP4 and HP2 displayed the highest growth index (37.8) and the highest triterpenoids concentration (46.57 mg∙g⁻¹), respectively. Although triterpenoid concentration was >2-fold higher in leaves than in petioles of C. asiatica plants, the accumulation of triterpenoids in petiole-derived HR cultures was 1.4-fold higher than that in leaf-derived HR cultures. Additionally, in both leaf- and petiole-derived HR cultures, terpenoid production was higher in HRs than in adventitious roots. These results demonstrate that the triterpenoid content in the explant does not affect the triterpenoid content in the resultant HRs. The HR culture of C. asiatica could be scaled up to enable the mass production of triterpenoids in bioreactors for the pharmaceutical and cosmetic industries. Full article

(This article belongs to the Special Issue Production of Secondary Metabolites In Vitro)

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