Antimicrobial and Antioxidant Metabolites From the Cultured Suspension Cells of Marchantia polymorpha L.

Abstract

Cell suspension culture is an attractive alternative source to wild plant for the production of novel biological metabolites. Hence, in this study, chemical investigation of cell suspension cultivated Marchantia polymorpha L. was performed, and led to the isolation of one new bis-bibenzyl (1), along with nine known analogues (2-10). Those chemical structures were elucidated based on the comprehensive analysis of NMR and MS data. The antioxidant and antibacterial effects of isolated components and crude extracts were evaluated, resulting in the identification of some antioxidant and antibacterial components. Meanwhile, Compare to that of wild grown M. polymorpha, the cell cultivated one was found to produce superior phenol yields, these constituents are of great importance for their antioxidant and antimicrobial activities. The studies conducted so far have established that the cell culture of M. polymorpha can be considered not only as a rich source of phenolic but as promising source of natural antioxidants and antibiotics as well, which is also expected to develop for better usage of this medicinal herb.

Keywords

cell suspension cultivation bis-bibenzyl analogs antioxidant activity antimicrobial activity

Introduction

The bryophytes are found all over the world, it was considered as the ancestor of land plants. There are around 23 000 species were reported and divided into 3 phyla including Bryophyta, Marchantiophyta, and Anthoceratophyta.¹ Among those bryophytes, lipophilic metabolites such as terpenoids, polyketides, and aromatic components are the major ones, which exhibited interesting biological activities.^2‐9

Marchantia polymorpha, known as a traditional Chinese herbal medicine used in the treatment of cuts, fractures, snake bites, burns, scalds, and open wounds. Phytochemical research declared that the major metabolites in M. polymorpha are polyphenols (bis-bibenzyls and flavonoids), long-chain polyunsaturated fatty acids (AA, EPA), and terpenoids.^10‐15 Among which especially the bis-bibenzyls exhibited significant antibacterial, antifungal, anti-inflammatory, and anti-oxidant properties^16‐18; AA and EPA are also commonly known for their beneficial roles in human health.¹⁵

Cell suspension culture is an attractive alternative source to wild plants for the production of secondary metabolites,^19‐23 it offers a clear production system to make continuous supply, even quality, and stable rates; meanwhile, it is possible to obtain novel components to that are not found in parent plants. In this study, chemical composition, total phenols and total flavonoids contents, and biological activity were investigated. The cell cultivated M. polymorpha was found to produce superior phenol yields and preferable biological activities, which can be explained by the better production of some effective constituents in artificial conditions, it was also supported by the HPLC fingerprint analyses. Furthermore, guided by antioxidant and antibacterial activity, phytochemical research of methanol-soluble fraction of cultivated M. polymorpha was investigated, a totally of 10 bis-bibenzyls derivatives were isolated, and structure elucidated (Figure 1). Herein, details of the cell cultivation, contents determination, chemical isolation, structural identification, and the results of the biological activities screening are described.

Figure 1.

Bis-bibenzyl derivatives (1-9) and bibenzyl (10) yield from cell suspension cultures of Marchantia polymorpha L.

Results and Discussion

Total Phenol and Flavonoid Contents Determination

Phenolics and flavonoids are important metabolites that exhibited antioxidant activity. Consequently, the total phenol contents (TPC) and total flavonoid contents (TFC) values were evaluated and presented. Clearly, a significant difference with respect to their TPC values has been found in methanol and petroleum extracts. As presented in Table 1, the higher TPC value was found in methanol extracts of cell cultivated M. polymorpha (ME, 271.23 ± 2.07 mg GAE/g extract), Followed by the methanol extracts of wild M. polymorpha (WM, 225.45 ± 1.48 mg GAE/g extract). On the other hand, the lowest TPC value was recorded for the petroleum extract cell cultivated M. polymorpha (CP, 11.15 ± 1.13 mg GAE/g extract).

Table 1.

The Content of Total Phenols and Flavonoids in Different Extracts.

Samples	TPC (mg GAE/g extract)	TFC (mg QE/g extract)
WP	18.33 ± 0.51	ND
WM	265.45 ± 1.48	0.31 ± 0.07
CP	11.15 ± 1.13	ND
CM	271.23 ± 2.07	ND

Note: Each value is the mean ± SD of 3 independent measurements and 95% confidence intervals; TPC, total phenols content; TFC, total flavonoids contents; GAE, gallic acid equivalents; QE, quercetin equivalents; WP, petroleum extract of wild M. polymorpha; WM, methanol extract of wild M. polymorpha; CP, petroleum extract of cultivated M. polymorpha; CM, methanol extract of cultivated M. polymorpha; ND, not detected.

Furthermore, the total flavonoid contents of crude extracts were also evaluated. However, none of the samples investigated exhibited a high TFC value, with the highest value found in methanol extracts of wild M. polymorpha was around 0.31 ± 0.07 mg QE/g extract. Moreover, no flavonoid metabolites were detected from the crude extract of cultivated M. polymorpha.

Structure Identification

Compound 1 was yielded as a white amorphous powder. Its molecular formula was determined as C₂₉H₂₆O₄ due to the (+)-HR-ESIMS data at m/z 439.1908 (calcd. for C₂₉H₂₇O₄, 439.1909, [M + H]⁺), indicative of 17° of unsaturation.

Four benzylic methylenes, 24 benzene ring carbon atoms, and 1 methyl group were determined based on the ¹H and ¹³C NMR spectrum, indicating that compound 1 was a bis-bibenzyl derivative. In the ¹H NMR spectrum, one AA'BB′ spin system at δ_H 6.75 (2H, d, J = 7.7 Hz, H-2/6), 6.93 (1H, d, J = 7.7 Hz, H-3), 6.83 (1H, d, J = 7.7 Hz, H-5) (ring A); one 12,3-trisubstituted benzene ring system at δ_H 6.98 (1H, brd, J = 7.9, 1.4 Hz, H-10), 7.18 (1H, t, J = 7.9 Hz, H-11), and 6.77 (1H, brd, J = 7.9 Hz, H-12) (ring B) were observed; as well as two 12,4-trisubstituted phenyl groups (AMX spin system) at δ_H 5.44 (1H, d, J = 1.6 Hz, H-3′), 6.66 (1H, dd, J = 8.0, 1.6 Hz, H-5′), and 6.81 (1H, d, J = 8.0 Hz, H-6′) for ring C, while 6.27 (1H, brs, H-10′), 6.29 (1H, d, J = 7.7 Hz, H-12′), 6.78 (1H, d, J = 7.7 Hz, H-13′) for ring D, respectively.

The linkage of rings A and B, rings C and D were determined by the HMBC experiment (Figure 2). Meanwhile, the biphenyl linkage between C-14 and C-14′ was also confirmed based on the HMBC correlations from H-13′ to C-14 (Figure 2). Thus, compound 1 was suggested to share the plgiochin skeleton. Comparison of the ¹H and ¹³C data with those reported plagiochin E led to the observation of an additional O-methyl at δ_H 3.75 (δ_C 55.3), which was deduced to be at C-1′ due to the HMBC connection from -OCH₃ to C-1′ (δ_C 152.8). Therefore, the chemical structure of 1 was determined as 7,10-Etheno-12,16-metheno-16H-dibenz[h,j]oxacyclooctadecin-1-methoxyl-13,20-diol-5,6,17,18-tetrahydro, and named plagiochin E 1′-O-methyl ether.

Figure 2.

Key HMBC and COSY connections of compound 1.

In addition to the aforementioned new compound, 9 known components including plagiochin G (2), marchantin A (3), marchantin B (4), marchantin F (5), marchantin D (6), marchantin K (7), marchantin E (8), perrottetin E (9), and lunularin (10) were isolated and identified. Their structures were finally elucidated by comparison of their NMR spectral data (presented in Supporting Information) with reported literature,^22‐27 as well as ESI-MS data.

Compound 1: White powder; (+)-ESIMS m/z439 [M + H]⁺; (+)-HR-ESIMS m/z 439.1921 (Calcd. for C₂₉H₂₇O₄, 439.1909). ¹H NMR (500 MHz, CD₃OD-d₄): δ_H 7.18 (1H, t, J = 7.8 Hz, H-11), 6.98 (1H, brd, J = 7.8 Hz, H-10), 6.93 (1H, d, J = 7.7 Hz, H-3), 6.83 (1H, d, J = 7.7 Hz, H-5), 6.81 (1H, d, J = 8.0 Hz, H-6′), 6.78 (1H, d, J = 7.7 Hz, H-13′), 6.77 (1H, brd, J = 7.8 Hz, H-12), 6.75 (1H, d, J = 7.7 Hz, H-2), 6.75 (1H, d, J = 7.7 Hz, H-6), 6.66 (1H, dd, J = 8.0, 1.6, H-5′), 6.29 (1H, d, J = 7.7 Hz, H-12′), 6.27 (1H, brs, H-10′), 5.44 (1H, d, J = 1.6 Hz, H-3′), 3.75 (3H, s, −OCH₃), 2.90 (1H, m, H-7a), 2.87 (2H, m, H-8), 2.74 (1H, m, H-7b), 2.65 (2H, m, H-7′), 2.65 (1H, m, H-8'a), 2.56 (1H, m, H-8'b); ¹³C NMR (125 MHz, CD₃OD-d₄): δ_C 155.4 (C-13), 154.5 (C-1), 155.0 (C-11′), 152.8 (C-1′), 148.8 (C-2′), 145.3 (C-9), 143.0 (C-9′), 141.6 (C-4), 134.2 (C-4′), 134.0 (C-13′), 130.5 (C-3), 130.1 (C-5), 129.5 (C-11), 126.6 (C-14), 123.1 (C-10), 123.1 (C-12′), 123.0 (C-2), 123.0 (C-6), 122.9 (C-5′), 121.7 (C-14′), 118.0 (C-3′), 117.8 (C-10′), 116.9 (C-6′), 114.0 (C-12), 55.3 (−OCH₃), 38.8 (C-8), 38.7 (C-8′), 37.9 (C-7′), 35.9 (C-7).

HPLC Fingerprint Analyses of Phenolics in Methanol Extracts between Wild and Cell Cultivated M. polymorpha

To determine the chemical propriety differences between wild and cell cultivated M. polymorpha, methanol extracted was prepared for HPLC analysis, using isolated compounds as reference standards. In HPLC analysis, compounds 1 to 10 were eluted as peaks with retention times at 39.38, 23.59, 31.44, 30.35, 24.72, 25.84, 28.91, 33.88, 36.73, and 41.03 min, respectively (Supplemental Figure S1). For compounds 1, 7, and 9, it was only detected in the cell suspension culture extracts. On the other hand, the content of compounds 3, 4, and 8 was much higher in cell suspension culture extracts, compared to that of natural growth one.

Antioxidant Activities

The DPPH and reducing power assays are easy and sensitive methods used in the evaluation of the antioxidative activity. Among which the DPPH assay has been commonly used as a tool for evaluating free radical scavenging activities of antioxidants, the characteristic absorption possessed and measured at 520 nm; while the reductant conversed the iron ion/ferricyanide complex to ferrous ion, and the remained iron ion can be monitored at 700 nm in reducing power assay.

Those results obtained from the DPPH assay are presented in Figure 3, the radical scavenging effects of isolated compounds on DPPH radicals were significant and concentration-dependent. Meanwhile, the reducing power results of tested samples are in agreement with the DPPH assay (Figure 4), all the tested samples exhibited moderate to significant activity. While methanol extracts were the most active ones, which showed comparable antioxidant effects to that of vitamin E in both assays.

Figure 3.

Radical scavenging activity (RSA) of tested samples (1-10: isolated components; CM: crude methanol extract of cell cultivated Marchantia polymorpha; PE: crude petroleum ether extract of cell cultivated M. polymorpha; WM: crude methanol extract of wild growth M. polymorpha; WP: crude petroleum ether extract of wild growth M. polymorpha), and standard antioxidants vitamin E. Values were expressed as mean ± SD (n = 3).

Figure 4.

Reducing power of tested samples (1-10, isolated components; CM, crude methanol extract of cell cultivated Marchantia polymorpha; PE, crude petroleum ether extract of cell cultivated M. polymorpha; WM, crude methanol extract of wild growth M. polymorpha; WP, crude petroleum ether extract of wild growth M. polymorpha), and standard antioxidants vitamin E. Values were expressed as mean ± SD (n = 3).

Antimicrobial Activities

To screen the antimicrobial activities of crude extract and isolated components, the in vitro antibacterial effect of crude extracts and pure components against human pathogenic bacteria is evaluated. The minimum inhibition concentration (MIC) values were presented in Tables 2 and 3. The methanol extracts exhibited interesting antibacterial activity against Staphylococcus aureus, Bacillus subtilis, and Listeria monocytogenes, as well as weak antibacterial activity against Escherichia coli. It declared that S. aureus and L. monocytogenes were the most sensitive bacterial strains to methanol extract. Alternatively, none of them exhibited an obviously antibacterial effect against Salmonella typhimurium.

Table 2.

Minimal Inhibitory Concentration (MIC, µg/mL) Values of Tested Samples Against Selected Microbial Strains.

Strains	Isolated components
Strains	1	2	3	4	5	6	7	8	9	10
S. aureus	512	–	128	128	128	128	128	128	128	–
B. subtilis	–	–	–	-	512	–	512	–	-	–
L. monocytogenes	512	–	–	512	–	–	–	–	512	–
S. typhimurium	–	–	–	–	–	–	–	–	–	–
E. coli	–	–	–	512	–	–	–	–	–	–
A. flavus	–	–	–	–	–	–	–	–	–	–
A. niger	–	–	–	–	–	–	–	–	–	–
C. albicans	128	256	–	256	512	512	128	–	–	–
T. mentagrophytes	–	–	–	512	512	–	–	–	256	–

Note: Bacterial strains: Staphylococcus aureus; Bacillus subtilis; Listeria monocytogenes; Salmonella typhimurium; and Escherichia coli, Fungal strains: Aspergillusflavus; Aspergillus niger; Candida albicans; Trichophyton mentagrophytes; PE, petroleum ether extract of cultivated M. polymorpha; ME, Methanol extract of cultivated M. polymorpha; “–”: No activity.

Table 3.

Minimal Inhibitory Concentration (MIC, µg/mL) Values for Crude Extract of Marchantia polymorpha on Selected Microbial Strains.

Bacterial strains	Nature		Cultivated
Bacterial strains	Petroleum	Methanol	Petroleum	Methanol
S. aureus	512	64	512	64
B. subtilis	–	512	–	256
L. monocytogenes	–	256	–	256
S. typhimurium	–	–	–	–
A. flavus	–	–	–	–
A. niger	–	–	–	–
C. albicans	–	256	–	64
T. mentagrophytes	–	256	–	128

Furthermore, the antifungal activity is also reported in Tables 2 and 3. Obviously, both extracts exhibited antifungal activity against selected fungi strains. The most effective antifungal activity was found for methanol extracts, which showed the best antifungal effect against Candida albicans and Trichophyton mentagrophytes, with MIC values at 64 and 128 µg/mL; respectively. Additionally, all the bis-bibenzyls (1-9) exhibited selected weak to moderate antifungal activity with MIC values from 64 to 512 µg/mL. From a structure–activity relationship analysis, it seems that the free hydroxyl moiety is necessary for the antifungal activity.

Materials and Methods

Plant Material

The fresh plant of M. polymorpha (100 g) was collected in the suburb of Chongqing province by one of the authors (T. Chen). Following a rinse with sterile distilled water, a small piece of M. polymorpha callus tissue was transferred into freshly prepared Murashige-Skoog's medium (MSK-2 medium),²⁸ which contained 2,4-dichlorophenoxyacetic acid (1 mg/L)²⁸ as phytohormone for cell growth, and then cultivated with continuous shaking (100 rev/min) in light at 25 °C (2000 Lux). Five days later, the cells were sub-cultured into a 1-L conical flask containing 500 mL of MSK-2 medium and then grown under the same condition for 4 weeks.

Extraction, Isolation, and Structure Elucidation

The cultured medium (15 000 mL) was filtered, and the cultivated plant cells were dried in vacuo and extracted by EtOH at room temperature for 3 days (1 L × 3 times), the crude extracts were further partitioned between petroleum ether and 90% methanol–water system (v/v) to give petroleum ether extract (CP, 4.3 g) and methanol extract (CM, 11.2 g), respectively. Meanwhile, about 80 g of fresh plants of M. polymorpha were extracted with petroleum at room temperature for 24 h (100 mL × 3) and filtered, the filtrate was further extracted with methanol under the same condition to give petroleum (WP, 0.4 g) and methanol extract (WM, 0.9 g).

Guided by biological activities, the methanol extract (11.2 g) was subjected to an MCI gel column and eluted with a gradient methanol–water system (from 50% to 100% methanol) to give 4 fractions. Fraction 2 was subjected to RP-HPLC and eluted with 42% MeCN (2.0 mL/min) to yield compounds 3 (914.3 mg), 5 (16.2 mg), and 6 (36.3 mg). Components 2 (31.7 mg), 4 (454.7 mg), and 7 (11.4 mg) were obtained from fraction 3 by eluting with 45% MeCN (2.0 mL/min). Compounds 1 (2.1 mg), 8 (74.5 mg), 9 (25.8 mg), and 10 (4.7 mg) were yielded from fraction 4 by eluting with 48% MeCN (2.0 mL/min). RP-HPLC experiment was carried out on a Shimadzu Liquid Chromatography LC-20AD using a Shimadzu UV/vis detector SPD-20A, with a YMC packed J'sphere ODS-H80 column (250 × 10 mm, 4 μm, 80 Å). The NMR data were recorded on Varian INOVA 500 spectrometers; meanwhile, MS and HR-ESIMS experiments were performed on the Finnigan LCQDECA instrument.

HPLC Fingerprint Analysis for Methanol Extracts of Wild and Cultivated M. polymorpha

Methanol extracts of the wild and cultivated M. polymorpha were dissolved in methanol to prepare sample solutions for HPLC analysis. Previously yield compounds in methanol were used as reference standards. The HPLC analysis was carried out on a YMC packed C18 column (5 µm, 4.6 mm × 250 mm, 12 nm), eluted with MeOH–H₂O system in linear-gradient (from 30% MeOH at the initial time to 90% MeOH at 90 min; flow rate, 0.6 mL/min) and detected at 254 nm.

Determination of Total Phenols and Total Flavonoids Contents

The total amount of phenols (TPC) in extracts was investigated as previously described.¹⁶ To 50 µL of prepared samples, 2.5 mL diluted Folin-Giocalteau's reagent, and 2 mL of sodium carbonate (7.5% v/v) were added and kept in a water bath (45 °C) for 15 min. The data was measured spectrophotometrically at 760 nm (Bio-Rad 680 microplate reader). The calibration curve was prepared by using a standard gallic acid solution from 20 to 1000 mg/L (r² = 0.998). The TPC values were presented as mg gallic acid equivalents per gram of extract (mg GAE/g extract).

The TFC values were determined as previously described.²⁹ In brief, 250 µL of extracts, 1.0 mL of distilled water, and 75 µL of 5% sodium nitrite were mixed and kept for 6 min, and then, 150 µL of aluminum trichloride (10% v/v) was added and further cultivated for another 5 min, and then, 0.5 mL of sodium hydroxide (1 M) was added. Finally, distilled water was added to make up to 2.5 mL and kept for another 15 min, the data was recorded immediately at 510 nm. The TFC was calculated using an equation obtained from the calibration curve of quercetin (r² = 0.999) and recorded as milligrams of quercetin equivalents per gram of extract (mg QE/g extract).

Antioxidant Activity Evaluation

DPPH assay: The DPPH (1,1-diphenyl-2-picrylhydrazyl) assay is commonly used to evaluate potential antioxidants to determine the antioxidant activity.³⁰ In brief, 0.1 mL of DPPH (150 µM) methanol solution was added to freshly prepared tested samples (0.1 mL), vigorously shaken and kept in darkness, 30 min later. The absorbance was recorded on a Bio-Rad 680 microplate reader at 520 nm. In this text, 0.1 mL of methanol was used as the blank control, while vitamin E was used as the positive standard. The RSA values were recorded:

RSA (%) = (1 – A_{S} / A_{B}) \times 100.

where A_S is the absorbance value of crude extract, A_B is the absorbance value of methanol.

Reducing power: The reducing power of each fraction was evaluated as previously described.³⁰ Briefly, tested samples (0.5 mL) of different concentrations were prepared and mixed with 0.5 mL of phosphate buffer (0.2 M, pH 6.6), 0.5 mL of potassium ferricyanide (1%), this mixture was initially cultivated at 50 °C for 20 min, and 0.5 mL of trichloroacetic acid (10%) were added. And then, the mixture was centrifugated at 3000 rev/min to remove the residue. Finally, 0.5 mL of the top layer, 0.5 mL distilled water, and 0.1 mL ferric chloride (1%) were mixed. The uniform mixture was kept at room temperature for 5 min and the data was determined at 700 nm (Shimadzu UV-2401 PC). In this study, antioxidant vitamin E was used as the positive control.

Antibiotic Susceptibility Tests

The antibacterial effect was simply evaluated by agar-well diffusion assay.³¹ Selected strains were cultivated in LB media overnight (37 °C) and diluted (1/100). Then, diluted bacterial culture (0.15 mL) was transferred to 100 mL autoclaved LB media (2.5 g powdered LB medium, and 1.5 g agar) to make plates (90 × 15 mm). Once the plates had been aseptically solidified, 10 μL of the test sample (10 mg/mL) were loaded, and further incubated at 37 °C for 18 h. Streptomycin (≥ 98%, 30 μg/disk) was used as a positive control. The inhibition zones surrounding the disks were recorded. Based on the aforementioned screening results, the MIC values were further evaluated.³² In brief, 20 μL selected bacterial cultures (105 CFU/mL) were added to a 96-well plate and mixed with 180 µL of prepared diluted samples. After 24 h incubation, the lowest concentration in which the bacterial strains do not visible growth was recorded and defined as the MIC values.

The antifungal activity was also investigated via a modified microdilution technique.³³ The fungal spores were harvested and diluted (1.0 × 10⁶ CFU/mL). The MIC values were determined by a serial dilution assay using 96-well microtiter plates. The crude extracts in broth medium and 1/10 diluted fungal inoculum were added to achieve different concentrations (0.1-10 mg/mL) and incubated at 28 °C. After 5 days, the lowest concentrations in which the fungal spores do not visible growth were defined as the MIC values.

Chemicals

All chemicals used in this study were purchased from Sigma.

Supplemental Material

sj-docx-1-npx-10.1177_1934578X221096172 - Supplemental material for Antimicrobial and Antioxidant Metabolites From the Cultured Suspension Cells of Marchantia polymorpha L.

Supplemental material, sj-docx-1-npx-10.1177_1934578X221096172 for Antimicrobial and Antioxidant Metabolites From the Cultured Suspension Cells of Marchantia polymorpha L. by Ya-yun Cai, Ting Chen and Jia-fu Cao in Natural Product Communications

Footnotes

Declaration of Conflicting Interests

The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Ethical Approval

Not applicable, because this article does not contain any studies with human or animal subjects.

Funding

The authors disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This work was supported by the Natural Science Foundation of Nantong City (grant no. JC 2020002).

Informed Consent

Not applicable, because this article does not contain any studies with human or animal subjects.

ORCID iD

Jia-fu Cao

Trial Registration

Not applicable, because this article does not contain any clinical trials.

Supporting Information

NMR and MS data were available as Supporting Information.

Supplemental Material

Supplemental material for this article is available online.

References

Wellman

Osterloff

Mohiuddin

. Fragments of the earliest land plants. Nature. 2003;425(6955):282‐285.

Asakawa

Ludwiczuk

. Chemical constituents of bryophytes: structures and biological activity. J Nat Prod. 2018;81(3):641‐660.

Asakawa

Nagashima

Ludwiczuk

. Distribution of bibenzyls, prenyl-bibenzyls, bis-bibenzyls, and terpenoids in the liverwort genus Radula. J Nat Prod. 2020;83(3):756‐769.

Asakawa

Ludwiczuk

Novakovic

Bukvicki

Anchang

. Bis-bibenzyls, bibenzyls, and terpenoids in 33 genera of the Marchantiophyta (Liverworts): structures, synthesis, and bioactivity. J Nat Prod. 2022;85(3):729‐762.

Yoshida

Toyota

Asakawa

. Isolation, structure elucidation, and chemical derivatization of a new cyclic bisbibenzyl dimer, Pusilatin E, from the liverwort Riccardia multifida subsp. decrescens. J Nat Prod. 1997;60(2):145‐147.

Quang

Hashhimoto

Tanaka

Takaoka

Asakawa

. Tyromycic acids B-E, new lanostane triterpenoids from the mushroom Tyromyces fissilis. J Nat Prod. 2004;67(2):148‐151.

Asakawa

Ludwiczuk

Nagashima

. Chemical constituents of bryophytes, chemical relationships between algae, bryophytes, and pteridophytes. In: Kinghorn

Falk

Kobayashi

, eds. Progress in the Chemistry of Organic Natural Products. Vol 95. Springer; 2013:1‐796.

Asakawa

. Chemical constituents of the Hepaticae. In: Herz

Grisebach

Kirby

, eds. Progress in the Chemistry of Organic Natural Products. Vol 42. Springer; 1982:1‐285.

Asakawa

. Chemical constituents of the Bryophytes. In: Herz

Kirby

Moore

Steglich

Tamm

, eds. Progress in the Chemistry of Organic Natural Products. Vol 65. Springer; 1995:1‐618.

10.

Fang

Guo

H-F

Lou

H-X

. Three new bibenzyl derivatives from the Chinese Liverwort Marchantia polymorpha L. Helv Chim Acta. 2007;90(4):748‐752.

11.

Gökbulut

Satilmiş

BatÇioğlu

ÇETİN

Şarea

. Antioxidant activity and luteolin content of Marchantia polymorpha L. Turk J Biol. 2012;36(4):381‐385.

12.

Jensen

JSRE

Omarsdottir

Thorsteinsdottir

Ogmundsdottir

Olafsdottir

. Synergistic cytotoxic effect of the microtubule inhibitor Marchantin A from Marchantia polymorpha and the Aurora kinase inhibitor MLN8237 on breast cancer cells in vitro. Planta Med. 2012;78(5):448‐454.

13.

Jensen

Omarsdottir

Bwalya

Nielsen

Tasdemir

Olafsdottir

. Marchantina A, a macrocyclic bisbibenzyl ether, isolated from the liverwort Marchantia polymorpha, inhibits protozoal growth in vitro. Phytomedicine. 2016;19(13):1191‐1195.

14.

Niu

J-B

Lou

H-X

. Antifungal bis[bibenzyls] from the Chinese liverwort Marchantia polymorpha L. Chem Biodivers. 2006;3(1):34‐40.

15.

Kajikawaa

Yamatoa

Sakaib

Fukuzawaa

Ohyamac

Kohchia

. Isolation and functional characterization of fatty acid D5-elongase gene from the liverwort Marchantia polymorpha L. FEBS Lett. 2006;580(1):149‐154.

16.

Mocan

Cri$an

Vlase

,Cri$an O, Vodnar DC, Ratia O, Gheldiu AM, Toiu A, Oprean R, Tilea I. Comparative studies on polyphenolic composition, antioxidant and antimicrobial activities of Schisandra chinensis leaves and fruits. Molecules. 2014;19(9):15162‐15179.

17.

J-B

Xie

C-F

Shi

Y-Q

Lou

H-X

. Water soluble constituents from the liverwort Marchantia polymorpha. Helv Chim Acta. 2007;90(11):2109‐2115.

18.

Saroya

. Herbalism, Phytochemistry and Ethnopharmacology. CRC Press Science Publishers Punjab, 2011; 286-293.

19.

Ravishankar

Bhyalakshmi

Ramachandra

. Production of food additives. In: Ramawat

Merillon

, eds. Biotechnology: Secondary metabolites. Oxford IBH; 1999:89‐110.

20.

Dornenburg

Knorr

. Cellular permeabilization of cultured plant tissues by high electric field pulses or ultra-high pressure for the recovery of secondary metabolites. Food Biotechnol. 1993;7(1):35‐48.

21.

Scragg

. Plant cell bioreactors. In: Stafford

Warren

, eds. Plant cell and tissue culture. Open Univ. Press; 1991:221‐239.

22.

Dornenburg

Knorr

. Production of the phenolic flavor compounds with cultured cells and tissues of Vanilla species. Food Biotechnol. 1996;10(1):75‐92.

23.

DiCosmo

Misawa

. Plant cell and tissue culture: alternatives for metabolite production. Biotechnol Adv. 1995;13(3):425‐435.

24.

Asakawa

Tori

Takikawa

Krishnamurty

Kantikar

. Cyclic bis (bibenzyls) and related compounds from the liverworts Marchantia polymorpha and Marchantia palmata. Phytochemistry. 1987;26(6):1811‐1816.

25.

Keseru

Nogradi

. Biosynthesis and molecular strain. A computational study on the conformation of cyclic bis(bibenzyl) constituents of liverwort species. Phytochemistry. 1992;31(5):1573‐1576.

26.

Tori

Masuya

Takikawa

Toyota

Asakawa

. The structure and NMR of macrocyclic bis(bibenzyls) isolated from liverworts. Tennen Yuki Kagobutsu Toronkai Koen Yoshishu. 1986;28:9‐16.

27.

Tori

Toyota

Harrison

. Total assignment of ¹H and ¹³C NMR spectra of Marchantins isolated from Liverworts and its application to structure determination of two new macrocyclic bisbibenzyls from Plagiochaama intermedium and Riccardia multifida. Tetrahedron Lett. 1985;26(39):4735‐4738.

28.

Yin

Wen

Lou

H-X

. Initiation of callus tissues and establishment of suspension culture of Marchantia polymorpha L. Cells. J. Shandong Univ. (Health Sciences). 2008;46(4):433‐437.

29.

Zhang

Yao

Duan

Wei

. Variation of essential oil content and antioxidant activity of Lonicera species in different sites of China. Ind Crop Prod. 2015;77(23):772‐779.

30.

Kumaran

Karunakaran

. Antioxidant and free radical scavenging activity of an aqueous extract of Coleus aromaticus. Food Chem. 2006;97(1):109‐114.

31.

Lehrer

Rosenman

Harwig

SSSL

Jackson

Eisenhauer

. Ultrasensitive assays for endogenous antimicrobial polypeptides. J Immunol Methods. 1991;137(2):167‐173.

32.

Ding

Liang

, et al. Activated production of silent metabolites from marine-derived fungus Penicillium citrinum. Fitoterapia. 2018;127:207‐211.

33.

Daouk

Dagher

Sattout

. Antifungal activity of the essential oil of Origanum syriacum L. J Food Protect. 1995;58(10):1147‐1149.

Supplementary Material

Please find the following supplemental material available below.

For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.

For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.

0.00 MB

0.45 MB