Next Article in Journal
Effect of Environmental and Anthropic Conditions on the Development of Solanum peruvianum: A Case of the Coastal Lomas, Lima-Peru
Previous Article in Journal
Stimulation of Arabidopsis thaliana Seed Germination at Suboptimal Temperatures through Biopriming with Biofilm-Forming PGPR Pseudomonas putida KT2440
Previous Article in Special Issue
Alkaloid Accumulation and Distribution within the Capsules of Two Opium Poppy (Papaver somniferum L.) Varieties
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Plants
Volume 13
Issue 19
10.3390/plants13192682
plants-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles thumb_up
...
Endorse textsms
...
Comment
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

Comparative Analysis of Volatile Compounds and Biochemical Activity of Curcuma xanthorrhiza Roxb. Essential Oil Extracted from Distinct Shaded Plants

by
Waras Nurcholis
1,2,*,
Rahmadansah Rahmadansah
2,
Puji Astuti
3,
Bambang Pontjo Priosoeryanto
4,
Rini Arianti
5,6,† and
Endre Kristóf
5,†
1
Tropical Biopharmaca Research Center, IPB University, Bogor 16151, Indonesia
2
Department of Biochemistry, Faculty of Mathematics and Natural Sciences, IPB University, Bogor 16680, Indonesia
3
Department of Biochemistry and Biomolecular Science, Faculty of Medicine, Universitas Tanjungpura, Pontianak 78124, Indonesia
4
Division of Veterinary Pathology, Faculty of Veterinary Medicine, IPB University, Bogor 16680, Indonesia
5
Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Debrecen, H-4032 Debrecen, Hungary
6
Universitas Muhammadiyah Bangka Belitung, Pangkalpinang 33684, Indonesia
*
Author to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Plants 2024, 13(19), 2682; https://doi.org/10.3390/plants13192682
Submission received: 3 September 2024 / Revised: 18 September 2024 / Accepted: 19 September 2024 / Published: 25 September 2024
(This article belongs to the Special Issue Secondary Metabolites in Plants)
Browse Figures
Review Reports Versions Notes

Abstract

:
The application of shade during plants’ growth significantly alters the biochemical compounds of the essential oil (EO). We aimed to analyze the effect of shade on the volatile compounds and biochemical activities of EO extracted from Curcuma xanthorrhiza Roxb. (C. xanthorrhiza) plants. Four shading conditions were applied: no shading (S0), 25% (S25), 50% (S50), and 75% shade (S75). The volatile compounds of EO extracted from each shaded plant were analyzed by gas chromatography–mass spectrometry. The antioxidant, antibacterial, and antiproliferative activities of EO were also investigated. We found that shade application significantly reduced the C. xanthorrhiza EO yield but increased its aroma and bioactive compound concentration. α-curcumene, xanthorrhizol, α-cedrene, epicurzerenone, and germacrone were found in EO extracted from all conditions. However, β-bisabolol, curzerene, curcuphenol, and γ-himachalene were only detected in the EO of S75 plants. The EO of the shaded plants also showed higher antioxidant activity as compared to unshaded ones. In addition, the EO extracted from S75 exerted higher antiproliferative activity on HeLa cells as compared to S0. The EO extracted from S0 and S25 showed higher antibacterial activity against Gram-positive bacteria than kanamycin. Our results suggest that shade applications alter the composition of the extractable volatile compounds in C. xanthorrhiza, which may result in beneficial changes in the biochemical activity of the EO.

1. Introduction

In their natural environment, plants typically grow in clusters, leading to inevitable mutual shading. These shading environments directly impact the light spectrum received by plants, typically characterized by the dominance of blue light (B), ranging between 400 and 500 nm and red (R) and far-red (FR) light between 600 and 700 nm and 700 and 750 nm, respectively. Additionally, there is a relatively low presence of ultraviolet-B light (UV-B) between 280 and 320 nm [1,2]. These light wavelengths are crucial for photosynthesis rates, thereby affecting plant growth and yield production [3,4,5,6]. The provision of shade, either natural or artificial, affects the amount and type of light received by plants through tailored shading, making this strategy valuable to mitigating global warming issues in crop production [7,8].
Several studies have demonstrated that shading significantly impacts both the production and composition of essential oil (EO) in various plants [9,10,11,12]. For instance, EO extracted from Ocimum basilicum L. cv. ‘Genovese’ under 50% blue net shading exhibited enhanced antimicrobial efficacy against Staphylococcus aureus (S. aureus), Escherichia coli (E. coli), and Proteus vulgaris (P. vulgaris) [12]. Furthermore, employing a 40% shade index for Anethum graveolens L. leads to increased EO yield and elevated activity against Klebsiella pneumoniae (K. pneumoniae) [13]. Understanding the influence of environmental factors on the production of secondary metabolites in medicinal plants is crucial due to their medical importance.
Curcuma xanthorrhiza Roxb. (C. xanthorrhiza), commonly known as Temulawak or Java turmeric, is a native herb from Indonesia with significant importance in traditional and herbal practices [14,15]. It plays a vital role in promoting the well-being of the Indonesian population through its various health benefits, including its antimicrobial and anticancer properties [16,17,18,19,20,21]. These therapeutic effects are attributed to the rich assortment of bioactive compounds found in its rhizomes, including terpenoids, phenols, flavonoids, saponins, alkaloids, and coumarins [14]. The rhizome, which serves as the source of active secondary metabolites such as curcuminoids and EOs, plays a crucial role in C. xanthorrhiza utilization and application for therapeutic purposes.
EOs comprise volatile and non-volatile compounds, which are secondary metabolites produced by various parts of plants [22]. Their production is influenced both by biotic and abiotic factors, including exposure to sunlight [23]. Modifying the light conditions through shading was shown to increase the percentage of EO production in thyme, marjoram, and oregano [24]. Moreover, shading may either enhance or diminish the production of specific secondary metabolites. For instance, the application of red-colored nets resulted in the highest percentage of patchoulol in Pogostemon cablin [11]. Similarly, employing 40% shading in Anethum graveolens L. enhanced the production of carvone but reduced the amount of limonene [13]. Nevertheless, the impact of shading on the EO composition of C. xanthorrhiza remains relatively unexplored to date.
Gas chromatography–mass spectrometry (GC-MS) has become a standard method in elucidating the components of EOs [25] and identifying metabolites in plant extracts [26,27,28]. In this study, we aimed to explore the chemical composition of C. xanthorrhiza EO in response to varying degrees of shading (0, 25, 50, and 75%) using GC-MS analysis. Chemometric analysis was employed to classify the shading levels of C. xanthorrhiza. Our investigation not only revealed the amount of the distinct volatile compounds but also evaluated the antioxidant, antibacterial, and antiproliferative properties of the EOs extracted from distinct shading conditions. This comprehensive examination aims to bridge gaps in our understanding of the EO composition of C. xanthorrhiza under different shading conditions, offering valuable insights into its potential as an antioxidant, antibacterial, and antiproliferative extract.

2. Materials and Methods

2.1. Chemicals

Ethanol (pro analysis), distilled water, HCl, BaCl2, H2SO4, ammonium acetate buffer, and trolox were purchased from Merck-Millipore (Darmstadt, Germany). 2,2-diphenyl-1-picrylhydrazyl (DPPH) powder was obtained from Himedia (Maharashtra, India). Ferric chloride (FeCl3) and 2,4,6-tripyridyl-s-triazine (TPTZ) were purchased from Sisco Research Laboratories Pvt. Ltd. (Maharashtra, India). S. aureus, E. coli, nutrient broth (NB), disc paper, kanamycin sulfate (KS), and muller hiton agar (MHA) were purchased from Sigma-Aldrich (St. Louis, MO, USA).

2.2. Sample Preparation and Shading Conditions

C. xanthorrhiza Roxb. varietas Cursina-III rhizomes with buds weighing 50–70 g were planted between January and September 2022 on open land within the Tropical Biopharmaca Cultivation Conservation, Unit Park of IPB University, located in Bogor, Indonesia, at an elevation of around 240 m above sea level, with geographic coordinates of 6°18′ 6°47′10 S and 106°23′45–107°13′30 W. This area has a wet tropical climate with rainfall with a range of temperature of 20–30 °C. All C. xanthorrhiza plants received the same conditions, including the type and amount of fertilizer, which contained urea at 250 kg/ha, SP-36, KCl each at 200 kg/ha, and 20 tons/ha of cow manure.
The plants were randomly grouped into 4 shade treatments: S0 (0%), S25 (25%), S50 (50%), and S75 (75%), with three replications for each group. The light grey-colored shades (AgroPro, PT. Gani Arta Dwitunggal, Padalarang, Indonesia) were composed of polyethylene-containing materials and configured as woven plastic nets. The density of the net was 66/127 holes/cm2. The percentage of shading represented the net density, which influenced the intensity of sunlight received by the plant. After 9 months, the rhizomes were harvested and immediately used for further analysis. Before processing, the plants were identified by an expert from the Tropical Biopharmaca Research Center, IPB University. Three kilograms of C. xanthorrhiza rhizomes from each shade condition was washed and chopped into 1–2 cm thickness for EO extraction, which was performed by using a simple steam distillation method. The samples were placed in distilled water for 4 h at 100–105 °C [28], and the yielded EO was used for further analysis.

2.3. Chemical Composition Analysis by Gas Chromatography–Mass Spectrometry (GC-MS)

The chemical composition of C. xanthorrhiza EO was analyzed by using Agilent Technologies 5977 GC-MS (Santa Clara, CA, USA). Three replicates of EO extraction were pooled and subjected to chemical compound analysis. The instrument was equipped with HP-5MS 5% PhenylMethylSilox (internal diameter 30 m × 250 µm with film thickness 0.25 µm) for the separation of the components. The GC oven temperature was initiated at 40 °C for 1 min, then gradually increased to 325 °C at a rate of 10 °C/min, and kept constant at 325 °C for 4 min. The relative amount of each identified volatile compound was quantified from the relative peak area of the chromatogram. Further confirmation of the chemical components was performed by comparing the retention time (RT) with the Willey 9 library database and PubChem data.

2.4. Antioxidant Activity Measurement

The following assays were carried out based on previously described methods [29] with modifications. Trolox was used as the standard, and the results were expressed as µmol of trolox equivalent (TE) per gram of dry weight (DW) or TE antioxidant capacity. DPPH and ferrous-reducing antioxidant activity (FRAP) assays were performed by using a modified nano-spectrophotometry method [30].

2.4.1. DPPH Assay

Briefly, 100 µL of 500 µg/mL EO solution from each shade condition of C. xanthorrhiza was added to 100 µL of 125 M DPPH solution (in pro-ethanol analysis) in a 96-well microplate (Biologix Europe GmbH, Hallbergmoos, Germany). Next, the samples were homogenized and incubated for 30 min in the dark at room temperature. The absorbance of the EO solution from each shade condition of C. xanthorrhiza was measured using a SPECTROstar Nano (BMG LABTECH, Ortenberg, Germany) at 515 nm.

2.4.2. FRAP Assay

Briefly, 10 µL of 1000 µg/mL C. xanthorrhiza EO from each shade condition and 300 µL of FRAP reagent (prepared by mixing acetate buffer at pH 3.6 with 10 M TPTZ solution (in 40 M HCl) and 20 µM FeCl3 (in distilled water) in a v/v/v ratio of 10:1:1) were placed in a 96-well microplate (Biologix Europe GmbH). The samples were then homogenized and incubated for 30 min in the dark at room temperature. The absorbance of the EO solution from each shade condition of C. xanthorrhiza was measured at a wavelength of 593 nm using SPECTROstar Nano (BMG LABTECH).

2.5. Antiproliferative Activity Analysis

All of the cell lines were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). Antiproliferative effects of the EO samples were measured on vero cells (CCL 81), a Michigan Cancer Foundation-7 (MCF-7) cell line for breast cancer (HTB 22), and a HeLa cell line for cervical cancer (CCL 2). Cells were seeded into 24-well plates at a density of 1 × 105 cells/well. Cells were grown in DMEM/F-12 medium supplemented with 10% FBS and C. xanthorrhiza EO from each shade condition at a final concentration of 4 µg/mL. Doxorubicin (Sigma-Aldrich) 0.2 µg/mL was applied as a positive control. The cell cultures were incubated at 37 °C and 5% CO2 for 3 days. Next, the cells were harvested for cell counting by using the trypan blue method. A total of 90 µL of the cell suspension and 10 µL trypan blue solution was added into a 96-well plate and homogenized. The cell suspension and trypan blue solution (Sigma-Aldrich) were then dropped onto the end of the hemocytometer, which was covered with a cover glass until all parts under the cover glass were filled with the solution. The cell counting was performed using a light microscope (Optika, Ponteranica, Italy) at 40× magnification. Round cells located in 25 boxes at the center of the hemocytometer were counted. The total number of cells comprised the living and dead cells. The living cells were colorless, while the dead cells were blue. The number of cells per ml, percent proliferation, and antiproliferation rate were calculated using the following formula:
% growth activity = ( Average number of cells by treatment Mean number of negative control cells ) × 100 %
% antiproliferative = 100% − (% growth activity)

2.6. Antibacterial Activity Analysis

Antibacterial activity analysis against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) was performed by using the disk diffusion method. A total of 10 µL of bacterial culture was briefly rejuvenated in 30 mL of NB medium supplemented with C. xanthorrhiza EO (4 µg/mL) and incubated for 24 h at 37 °C. The bacterial culture was streaked onto the media (MHA) and incubated for 24 h. The clear zone around the disc was measured using a caliper to assess the antibacterial activity.

2.7. Statistical Analysis and Figure Preparation

All measured values are expressed as mean ± standard deviations (SDs) for the number of independent repetitions indicated. Statistical analysis was carried out using one-way ANOVA followed by Tukey’s post hoc test. GraphPad Prism 9 (GraphPad Software, San Diego, CA, USA) was used for figure preparation and statistical analysis.

3. Results

3.1. Shade Results in Strong Aroma, Concentrated C. xantorrhiza EO, and Decreased Extract Yield

C. xanthorrhiza EO was produced from mature and thick rhizomes by using water-steam distillation (hydrodistillation) (Figure 1a). We found that shade enhanced the aroma and concentration of the yielded C. xanthorrhiza EO in an intensity-dependent manner (Figure 1a). No shade (S0) and 25% shade (S25) yielded a light yellow EO with less aroma. On the other hand, 50% (S50) and 75% shade (S75) yielded a more concentrated C. xanthorrhiza EO with a dark brown color and stronger aroma. Shade treatment decreased the yield (% v/w) of EO in an intensity-dependent manner (Figure 1b). These data suggested that shade treatment on plants resulted in a more concentrated but lower yield of C. xanthorrhiza EO.

3.2. Sesquiterpenes Are the Major Chemical Compounds in C. xanthorrhiza EO

Next, we analyzed the chemical composition of C. xanthorrhiza EO by GC-MS. Identification of the chemical compounds is essential to predict the biological activity or potential medical benefits that can be provided by the plant extract. We detected 63 chemical compounds that belong to various groups, such as monoterpenes, carboxylic ester, dialkyl phosphate, diterpenoid, fatty alcohol, organosiloxane, triterpenes, and sesquiterpenes (Figure 2a). Our results showed that sesquiterpenes comprise 75% of total chemical compounds, which were detected by GC-MS, in C. xanthorrhiza EO extracted from four different shade conditions (Figure 2a and Table 1). α-curcumene, xanthorrhizol, α-cedrene, epicurzerenone, and germacrone were the chemical compounds with the highest amount (4–9%) and found in EO extracted from all shades (Figure 2b and Table 1). Several sesquiterpenes compounds, such as β-bisabolol, curzerene, curcuphenol, and γ-himachalene, were only detected in C. xanthorrhiza EO extracted from the S75 group (Figure 2b and Table 1). Our results showed that modifying the environmental condition by installing shade altered the abundance of chemical compounds, especially sesquiterpenes, which may contribute to the beneficial biological activities of C. xanthorrhiza EO.

3.3. Shade Enhanced the Antioxidant Activity of C. xanthorrhiza EO

Secondary metabolites can be attributed to the antioxidative activity of EOs. We observed that C. xanthorrhiza EO contains various active compounds and that S75 increased the abundance of several sesquiterpenes. Next, we analyzed the antioxidant activity of C. xanthorrhiza EO from each shade condition by using two different radical scavenging methods, DPPH and FRAP. Our results showed that S25 and S75 increased the free radical scavenging activity of C. xanthorrhiza EO as compared to S0 measured by the DPPH assay (Figure 3, left panel). On the other hand, S75 showed increased antioxidant activity measured by the FRAP assay as compared to S0 (Figure 3, right panel). Our results suggested that the shade enhanced the antioxidant activity of C. xanthorrhiza EO, which may provide benefits as a pharmacological agent.

3.4. Condition of 75% Shade Elevates the Antiproliferative Activity of C. xanthorrhiza EO in HeLa Cells

Active compounds, such as sesquiterpenes, are potential pharmaceutical agents for the treatment of cardiovascular diseases and cancers. First, we checked the antiproliferative activity of the extracts in non-cancerous vero cells, which originated from kidney epithelial cells of an African green monkey. We found that the EO extracted from shaded plants showed a lower antiproliferative activity on vero cells as compared to doxorubicin (Figure 4, left panel). However, no difference was observed between each shade condition (Figure 4, left panel). Next, we investigated the antiproliferative activity of C. xanthorrhiza EO from each shade condition by calculating the proliferation rates of the cancer cell lines MCF-7 and HeLa (Figure 4, middle and right panels, respectively). Our results showed that EOs from all shades exerted a comparable antiproliferative activity in vitro in both MCF-7 and HeLa cells as compared to doxorubicin (Figure 4). C. xanthorrhiza EO from S75 showed higher antiproliferative activity as compared to S0 in HeLa cells (Figure 4, right panel). A higher abundance of sesquiterpenes in S75 may contribute to the more effective antiproliferative activity, especially in HeLa cells.

3.5. C. xanthorrhiza EO Exhibits Antibacterial Activity

Next, we also evaluated antibacterial activity of C. xanthorrhiza EO by using the disk diffusion method. S. aureus (Gram-positive) and E. coli (Gram-negative) were used for the antibacterial evaluation, in which KS was administered as the positive control. Our results showed that C. xanthorrhiza EO exerted antibacterial activity at a comparable level as KS (Figure 5). The antibacterial test against S. aureus showed that the EO from S0 and S25 possessed higher antibacterial activity as compared to KS (Figure 5, left panel), while the EO from S50 and S75 had lower antibacterial activity as compared to S0 against S. aureus (Figure 5, left panel). On the other hand, C. xanthorrhiza EO from S50 showed lower antibacterial activity against E. coli as compared to KS (Figure 5, right panel). These results suggest that C. xanthorrhiza EO from S0 and S25 exerted a stronger antibacterial effect on Gram-positive bacteria as compared to KS but not on Gram-negative bacteria.

4. Discussion

C. xanthorrhiza, or ’temulawak’, is an indigenous Indonesian plant characterized by a pseudostem and typically reaches a height of 1.29–2.50 m [14,31]. In Indonesia, C. xanthorrhiza is often cultivated together with other plants, such as Zea mays, Tectona grandis, Hevea brasiliensis, Azadirachta excelsa, and Albizia chinensis, creating a shaded environment conducive to its growth [32,33,34,35,36]. Our study demonstrated a negative correlation between shading density and the yield of C. xanthorrhiza EO. A previous study with Chinese wild ginger (Asarum heterotropoides Fr. Schmidt var. mandshuricum (Maxim) Kitag.) reported that unshaded plants, which received full sunlight exposure, had higher yielded extract, including EOs, as compared to unshaded ones [37].
C. xanthorrhiza is a phototrophic organism, relying on light-dependent photosynthesis for its metabolic process, including the production of primary and secondary metabolites. Photosynthetic efficiency is influenced by CO2 concentration, transpiration, photosynthetic rate, and CO2 intake through stomata (stomatal conductance) [38,39]. Our previous study reported that shading did not affect the quantity of C. xanthorrhiza plant tillers; however, 75% shading significantly decreased the rhizome biomass [40]. Shading has been found to downregulate the expression of NADPH.quinone oxireductase (NQO), a protein essential for cellular respiration and CO2 intake, thereby reducing the photosynthetic capacity and stomatal index [41,42]. The highest yield of EO production in our S0 treatment likely occurred due to full light exposure, which elevated the stomatal index and subsequently increased the rate of photosynthesis [43]. In line with our results, a reduction in light intensity (25% shading) has also been shown to decrease the EO production of Rosmarinus officinalis L. by up to 43% [44].
Our study indicated that shading altered the secondary metabolites of C. xanthorrhiza EO. Terpenoid sesquiterpenes dominated the volatile compound identification; however, several sesquiterpenes were found only in EO extracted from 75% shaded plants. Reduced photosynthesis due to shading condition did not inhibit the secondary metabolite production. A previous study reported that the production and accumulation of volatile compounds are affected by the developmental stage of the plants [45,46]. In our study, the rhizomes were harvested from matured plants after 9 months of planting; therefore, the bioactive compounds could be produced and accumulated in the rhizomes during the plant’s growth [47]. Different spectra and intensities of light synergistically influence the chemical composition of many plants [9,13,24,48,49,50]. The variation of secondary metabolite content in C. xanthorrhiza EO extracted from different shade conditions resulted in different antioxidant activities measured in this study. The 75% shading condition significantly elevated the antioxidant activity assessed by both DPPH and FRAP analyses. The condition with 25% shade showed elevated antioxidant activity only measured by the DPPH test, which is more suitable for hydrophobic samples, such as EOs, whereas FRAP is more optimal for hydrophilic samples [51]. Therefore, the DPPH test was more sensitive to the antioxidant activity of EOs. Although 75% shading produced the lowest EO yield, the variety of its secondary metabolite content was in a wider range, in addition to its stronger color and aroma.
In addition to light, other external factors such as temperature, CO2 concentration, and moisture can also affect the composition of both primary and secondary metabolites of plants [52]. The response of plants to climate change may alter the active compound composition that is attributed to the pharmaceutical properties of the plants. The plants from which EOs were extracted were planted at temperatures ranging from 20 to 30 °C, which support healthy growth without exposing the plants to cold or heat stress. Increased CO2 levels can also affect the plants’ growth and development and elevate biomass production. Plants growing at elevated CO2 levels typically exhibit reduced nutritional quality and nitrogen and protein content due to enhanced dilution of nutrients [52]. However, the response can be different depending on the duration of the exposure and other environmental factors. Another study conducted using C. xanthorrhiza (WP.5818) obtained from the market reported that the volatile compounds of the extracted EO were dominated by monoterpenes [53]. The difference in the environmental factors may cause this discrepancy with our study, which showed that sesquiterpenes built 75% of the volatile compounds.
Numerous studies have shown a positive correlation between antioxidant activity and antiproliferative effects against various cancers [54,55,56,57,58]. In association with the currently reported data, our previous study found that C. xanthorrhiza EO did not affect the proliferation of vero cells irrespective of the concentration, indicating the selectivity of the EO [59]. We also examined the effect of the EO samples on two cancer cell lines: MCF-7 for breast cancer and HeLa for cervical cancer. The 75% shading condition resulted in higher antiproliferative activity of the EO in HeLa cells compared to the unshaded condition. In our study, 75% shading stimulated the production of more chemical compounds, such as β-bisabolol, curzerene, curcuphenol, and γ-himachalene, which have been reported to possess anticancer properties in several studies [60,61,62,63,64]. Curzerene (C15H20O), a sesquiterpene commonly found in Curcuma rhizomes, was shown to suppress the proliferation of glioblastoma, melanoma, and human lung adenocarcinoma cells [62,63,65]. Curzerene was found to downregulate the expression of glutathione S-transferase (GST) A1 and A4 at the mRNA level, leading to apoptosis by preventing DNA damage during replication in the G0/G1 phase [62,63]. Additionally, curzerene inhibited cell proliferation by suppressing the activation of the mammalian target of the rapamycin (mTOR) pathway [62].
Bisabolol, which consists of α-bisabolol and β-bisabolol, is an active monocyclic sesquiterpene alcohol known for its anti-inflammatory and skin-soothing effects [66]. Research on β-bisabolol is less extensive than on its isomer, α-bisabolol. Several studies reported that β-bisabolol has similar anti-inflammatory properties, suggesting comparable pharmacological potential [67]. The α-bisabolol exhibits antiproliferative and cytotoxic effects, leading to apoptosis of various cancer cell lines [68,69,70,71]. These apoptotic effects are stimulated via the inhibition of the transducer and activator of transcription 3 (STAT-3), a transcription factor crucial in tumorigenesis and cancer progression [72]. Similarly, β-bisabolol from Semenovia suffruticosa has been demonstrated to exert cytotoxic properties against several types of cancer cells [73]. Curcuphenol and γ-himachalene, compounds found in C. xanthorrhiza grown under 75% shading, are also associated with apoptotic mechanisms in cancers. Curcuphenol induces apoptosis by enhancing the expression of caspase-3 and antigen processing and presentation machinery (APM), leading to a stronger immune response against metastatic tumor cells [64,74,75]. In addition, the anticancer properties of γ-himachalene may result from its cytotoxic effects [76,77,78].
Although sesquiterpenes, such as γ-himachalene and bisabolol, were identified in plants subjected to 75% shading, we did not find any noticeable change in the antibacterial properties of the corresponding EO [79,80,81,82]. Interestingly, our investigation found that the antibacterial efficacy was pronounced in C. xanthorrhiza plants cultivated under full sun exposure and with only 25% shading, particularly against S. aureus, a Gram-positive bacterium. Through comprehensive GC-MS analysis, we observed that these treatments exhibited less complex secondary metabolic profiles compared to denser shading conditions (50% and 75%). These results suggested that the observed antibacterial effect may be attributed to the presence of α-curcumene and α-cedrene, compounds found in higher concentrations in plants grown under full sun exposure or with 25% shading as compared to those subjected to 50% or 75% shading. Various studies have demonstrated a positive correlation between α-cedrene concentrations and the inhibition of S. aureus growth [83,84]. Additionally, α-curcumene has been shown to be effective against Gram-positive bacteria [85,86,87] by disrupting their cell membrane, leading to bacterial cell death [88].

5. Conclusions

The study highlights the significant impact of shading on the yield, composition, and bioactive properties of C. xanthorrhiza Roxb. EO. We found that increased shading density negatively correlated with EO yield, and full sunlight exposure enhanced its production due to increased photosynthetic efficiency. Shading also influenced secondary metabolite profiles. The 75% shading condition significantly boosted the antioxidant activity despite resulting in the lowest yield. This condition produced a broader variety of secondary metabolites, contributing to stronger color and aroma. Furthermore, EO from plants under 75% shading showed higher antiproliferative activity against HeLa cancer cells, attributed to compounds like β-bisabolol, curzerene, curcuphenol, and γ-himachalene. Interestingly, while sesquiterpenes, such as γ-himachalene and bisabolol, were prevalent under 75% shading, they lacked notable antibacterial properties, whereas plants under full sun and 25% shading exhibited strong antibacterial effects against S. aureus, linked to higher concentrations of α-curcumene and α-cedrene. Our findings highlight the importance of optimal shading conditions in maximizing the therapeutic potential of C. xanthorrhiza, paving the way for its broader applications in the pharmaceutical and healthcare sectors.

Author Contributions

Conceptualization, W.N.; methodology, R.R. and B.P.P.; software, P.A. and R.A.; validation, R.R. and W.N.; formal analysis, W.N.; investigation, W.N. and R.R.; resources, W.N. and B.P.P.; data curation, R.R. and R.A.; writing—original draft preparation, W.N., P.A., and R.A.; writing—review and editing, R.A. and E.K.; visualization, R.A.; supervision, W.N. and E.K.; project administration, W.N.; funding acquisition, W.N. and E.K. All authors have read and agreed to the published version of the manuscript.

Funding

This research was funded by the PDUPT IPB University research program (1925/IT3.L1/PN/2021) from the Ministry of Research, Technology and Higher Education of the Republic of Indonesia. R.A. and E.K. were supported by the National Research, Development and Innovation Office (NKFIH- PD146202 and FK145866) of Hungary and the University of Debrecen Program for Scientific Publication.

Data Availability Statement

The original contributions presented in the study are included in the article, and further inquiries can be directed to the corresponding author/s.

Conflicts of Interest

The authors declare no conflicts of interest.

References

  1. Fiorucci, A.S.; Fankhauser, C. Plant Strategies for Enhancing Access to Sunlight. Curr. Biol. 2017, 27, R931–R940. [Google Scholar] [CrossRef] [PubMed]
  2. Martinez-Garcia, J.F.; Rodriguez-Concepcion, M. Molecular mechanisms of shade tolerance in plants. New Phytol. 2023, 239, 1190–1202. [Google Scholar] [CrossRef] [PubMed]
  3. Tang, X.L.; Jiang, J.; Jin, H.P.; Zhou, C.; Liu, G.Z.; Yang, H. Effects of shading on chlorophyll content and photosynthetic characteristics in leaves of Phoebe bournei. Ying Yong Sheng Tai Xue Bao/J. Appl. Ecol. 2019, 30, 2941–2948. [Google Scholar] [CrossRef]
  4. De Souza, A.P.; Burgess, S.J.; Doran, L.; Hansen, J.; Manukyan, L.; Maryn, N.; Gotarkar, D.; Leonelli, L.; Niyogi, K.K.; Long, S.P. Soybean photosynthesis and crop yield are improved by accelerating recovery from photoprotection. Science 2022, 377, 851–854. [Google Scholar] [CrossRef] [PubMed]
  5. Shao, J.; Li, G.; Li, Y.; Zhou, X. Intraspecific responses of plant productivity and crop yield to experimental warming: A global synthesis. Sci. Total Environ. 2022, 840, 156685. [Google Scholar] [CrossRef]
  6. Li, T.; Liu, L.N.; Jiang, C.D.; Liu, Y.J.; Shi, L. Effects of mutual shading on the regulation of photosynthesis in field-grown sorghum. J. Photochem. Photobiol. B. 2014, 137, 31–38. [Google Scholar] [CrossRef]
  7. Lara-Estrada, L.; Rasche, L.; Schneider, U.A. Exploring the cooling effect of shading for climate change adaptation in coffee areas. Clim. Risk Manag. 2023, 42, 100562. [Google Scholar] [CrossRef]
  8. Tiffon-Terrade, B.; Simonneau, T.; Caffarra, A.; Boulord, R.; Pechier, P.; Saurin, N.; Romieu, C.; Fumey, D.; Christophe, A. Delayed grape ripening by intermittent shading to counter global warming depends on carry-over effects and water deficit conditions. OENO One 2023, 57, 71–90. [Google Scholar] [CrossRef]
  9. Viana, A.J.S.; Alves de Carvalho, A.; Alves de Assis, R.M.; Mendonça, S.C.; Rocha, J.P.M.; Pinto, J.E.B.P.; Bertolucci, S.K.V. Impact of Colored Shade Nets on Biomass Production, Essential Oil Composition and Orientin and Isoorientin Content in Lippia gracilis Schauer. Chem. Biodivers. 2023, 20, e202300809. [Google Scholar] [CrossRef]
  10. Lima, V.A.; Pacheco, F.V.; Avelar, R.P.; Alvarenga, I.C.A.; Pinto, J.E.B.P.; Alvarenga, A.A.D. Growth, photosynthetic pigments and production of essential oil of long-pepper under different light conditions. An. Acad. Bras. Cienc. 2017, 89, 1167–1174. [Google Scholar] [CrossRef]
  11. Ribeiro, A.S.; Ribeiro, M.S.; Bertolucci, S.K.; Bittencourt, W.J.; Carvalho, A.A.D.; Tostes, W.N.; Alves, E.; Pinto, J.E. Colored shade nets induced changes in growth, anatomy and essential oil of Pogostemon cablin. An. Acad. Bras. Cienc. 2018, 90, 1823–1835. [Google Scholar] [CrossRef] [PubMed]
  12. Ilić, Z.S.; Milenković, L.; Šunić, L.; Tmušić, N.; Mastilović, J.; Kevrešan, Ž.; Stanojević, L.; Danilović, B.; Stanojević, J. Efficiency of basil essential oil antimicrobial agents under different shading treatments and harvest times. Agronomy 2021, 11, 1574. [Google Scholar] [CrossRef]
  13. Milenković, L.; Ilić, Z.S.; Stanojević, L.; Danilović, B.; Šunić, L.; Kevrešan, Ž.; Stanojević, J.; Cvetković, D. Chemical Composition and Bioactivity of Dill Seed (Anethum graveolens L.) Essential Oil from Plants Grown under Shading. Plants 2024, 13, 886. [Google Scholar] [CrossRef]
  14. Rahmat, E.; Lee, J.; Kang, Y. Javanese Turmeric (Curcuma xanthorrhiza Roxb.): Ethnobotany, Phytochemistry, Biotechnology, and Pharmacological Activities. Evid.-Based Complement. Altern. Med. 2021, 2021, 9960813. [Google Scholar] [CrossRef]
  15. Syamsudin, R.A.M.R.; Perdana, F.; Mutiaz, F.S. Tanaman temulawak (Curcuma xanthorrhiza Roxb.) sebagai obat tradisional. J. Ilm. Farm Bahari. 2019, 10, 51–65. [Google Scholar] [CrossRef]
  16. Septama, A.W.; Tasfiyati, A.N.; Kristiana, R.; Jaisi, A. Chemical profiles of essential oil from Javanese turmeric (Curcuma xanthorrhiza Roxb.), evaluation of its antibacterial and antibiofilm activities against selected clinical isolates. S. Afr. J. Bot. 2022, 146, 728–734. [Google Scholar] [CrossRef]
  17. Suniarti, D.F.; Puspitawati, R.; Yanuar, R.; Herdiantoputri, R.R. Curcuma xanthorrhiza Roxb. an Indonesia native medicinal plant with potential antioral biofilm effect. In Focus on Bacterial Biofilms; IntechOpen: London, UK, 2022. [Google Scholar]
  18. Yasni, S.; Imaizumi, K.; Sugano, M. Effects of an Indonesian medicinal plant, Curcuma xanthorrhiza Roxb., on the levels of serum glucose and triglyceride, fatty acid desaturation, and bile acid excretion in streptozotocin-induced diabetic rats. Agric. Biol. Chem. 1991, 55, 3005–3010. [Google Scholar] [CrossRef]
  19. Sylvester, W.; Son, R.; Lew, K.; Rukayadi, Y. Antibacterial activity of Java turmeric (Curcuma xanthorrhiza Roxb.) extract against Klebsiella pneumoniae isolated from several vegetables. Int. Food Res. J. 2015, 22, 1770–1776. [Google Scholar]
  20. Simamora, A.; Timotius, K.H.; Yerer, M.B.; Setiawan, H.; Mun’im, A. Xanthorrhizol, a potential anticancer agent, from Curcuma xanthorrhiza Roxb. Phytomedicine. 2022, 105, 154359. [Google Scholar] [CrossRef]
  21. Musfiroh, I.; Geganaputra, A.; Diantini, A.; Susilawati, Y.; Muchtaridi, M. Antiproliferation Assay of Essential Oil of Curcuma Rhizoma (Curcuma xanthorrhiza Roxb.) Against P388 Leukemia Cell. Indones. J. Pharm. Sci. Technol. 2020, 7, 100–106. [Google Scholar] [CrossRef]
  22. Altemimi, A.; Lakhssassi, N.; Baharlouei, A.; Watson, D.G.; Lightfoot, D.A. Phytochemicals: Extraction, Isolation, and Identification of Bioactive Compounds from Plant Extracts. Plants 2017, 6, 42. [Google Scholar] [CrossRef] [PubMed]
  23. Suzuki, N.; Rivero, R.M.; Shulaev, V.; Blumwald, E.; Mittler, R. Abiotic and biotic stress combinations. New Phytol. 2014, 203, 32–43. [Google Scholar] [CrossRef] [PubMed]
  24. Milenković, L.; Ilić, Z.S.; Šunić, L.; Tmušić, N.; Stanojević, L.; Stanojević, J.; Cvetković, D. Modification of light intensity influence essential oils content, composition and antioxidant activity of thyme, marjoram and oregano. Saudi J. Biol. Sci. 2021, 28, 6532–6543. [Google Scholar] [CrossRef] [PubMed]
  25. Jabbar, M.; Ghorbaniparvar, H. Use of GC-MS combined with resolution methods to characterize and to compare the essential oil components of green and bleached cardamom. Int. J. Res. Chem. Environ. 2015, 5, 76–85. [Google Scholar]
  26. Ghimire, B.K.; Yoo, J.H.; Yu, C.Y.; Chung, I.M. GC–MS analysis of volatile compounds of Perilla frutescens Britton var. Japonica accessions: Morphological and seasonal variability. Asian Pac. J. Trop. Med. 2017, 10, 643–651. [Google Scholar] [CrossRef]
  27. Ukwubile, C.; Ahmed, A.; Katsayal, U.; Ya’u, J.; Mejida, S. GC-MS analysis of bioactive compounds from Melastomastrum capitatum (Vahl) Fern. leaf methanol extract: An anticancer plant. Sci. Afr. 2019, 3, e00059. [Google Scholar] [CrossRef]
  28. Nurcholis, W.; Khumaida, N.; Bintang, M.; Syukur, M. GC-MS analysis of rhizome ethanol extracts from Curcuma aeruginosa accessions and their efficiency activities as anticancer agent. Biodiversitas J. Biol Divers. 2021, 22, 1179–1186. [Google Scholar] [CrossRef]
  29. Nurcholis, W.; Alfadzrin, R.; Izzati, N.; Arianti, R.; Vinnai, B.Á.; Sabri, F.; Kristóf, E.; Artika, I.M. Effects of methods and durations of extraction on total flavonoid and phenolic contents and antioxidant activity of java cardamom (Amomum compactum Soland Ex Maton) fruit. Plants 2022, 11, 2221. [Google Scholar] [CrossRef]
  30. Arista, R.; Priosoeryanto, B.; Nurcholis, W. Total Phenolic, Flavonoids Contents, and Antioxidant Activities in The Stems and Rhizomes of Java Cardamom as Affected by Shading and N Fertilizer Dosages. Üzüncü Il Üniversitesi Tarım Bilim Derg. 2023, 33, 29–39. [Google Scholar] [CrossRef]
  31. Ma’tan, M.E.; Pinaria, A.G.; Kaligis, J.B.; Watung, J.F.; Paat, F.J.; Pioh, D.D. Plant Morphology and Analysis of Yellow Temulawak Curcumin (Curcuma xanthorrhiza Roxb.) in the Kinilow Village. J. Agroekoteknol. Terap. 2022, 3, 455–463. [Google Scholar] [CrossRef]
  32. Murdiono, W.E.; Nihayati, E.; Azizah, N. Peningkatan produksi temulawak (Curcuma xanthorrhiza) pada berbagai macam pola tanam dengan jagung (Zea mays). J. Hortik. Indones. 2016, 7, 129–137. [Google Scholar] [CrossRef]
  33. Nihayati, E.; Rizqullah, D.R.B.; Widaryanto, E. Strategy to improve temulawak (Curcuma xanthorrhiza) yields and quality under teak trees (Tectona grandis). J. Hortik. Indones. 2021, 12, 81–88. [Google Scholar] [CrossRef]
  34. Devy, L.; Tanjung, A.; Chotimah, S. Keragaman Genetik dan Korelasi Antar Karakter Agronomis Temulawak (Curcuma xanthorrhiza Roxb.) di Bawah Naungan Tegakan Karet. J. Agrotek. Indones. Indones. J. Agrotech. 2021, 6, 34–43. [Google Scholar]
  35. Arif, M.A.; Hartoyo, A.P.P.; Wijayanto, N.; Naufal, H. The potential of Curcuma longa, Curcuma xanthoriza and Centella asiatica in Silvofarmaka System Based on Melia azedarach and Azadirachta excelsa. In IOP Conference Series: Earth and Environmental Science; IOP Publishing: Bristol, England, UK, 2019; Volume 394. [Google Scholar] [CrossRef]
  36. Purnomo, D.; Budiastuti, M.S.; Sakya, A.T.; Cholid, M.I. The potential of turmeric (Curcuma xanthorrhiza) in agroforestry system based on silk tree (Albizia chinensis). In IOP Conference Series: Earth and Environmental Science; IOP Publishing: Bristol, England, UK, 2018; Volume 142. [Google Scholar] [CrossRef]
  37. Wang, Z.; Xiao, S.; Wang, Y.; Liu, J.; Ma, H.; Wang, Y.; Tian, Y.; Hou, W. Effects of light irradiation on essential oil biosynthesis in the medicinal plant Asarum heterotropoides Fr. Schmidt var. mandshuricum (Maxim) Kitag. PLoS ONE 2020, 15, e0237952. [Google Scholar] [CrossRef]
  38. Liang, Y.F.; Yi, J.N.; Wang, K.C.; Xue, Q.; Sui, L. Response of growth and photosynthetic characteristics of Polygonatum cyrtonema to shading conditions. Zhongguo Zhong Yao Za Zhi Zhongguo Zhongyao Zazhi China J. Chin. Mater. Med. 2019, 44, 59–67. [Google Scholar]
  39. Shan, Z.; Luo, X.; Wei, M.; Huang, T.; Khan, A.; Zhu, Y. Physiological and proteomic analysis on long-term drought resistance of cassava (Manihot esculenta Crantz). Sci Rep. 2018, 8, 17982. [Google Scholar] [CrossRef]
  40. Murwani, Z.A.; Artika, I.M.; Syaefudin, S.; Nurcholis, W. Evaluation of Growth, Chlorophyll Content, and Photosynthesis Rate of Curcuma xanthorrhiza with Different Shade Levels. Curr. Appl. Sci. Technology. 2024, 24, e0256871. [Google Scholar] [CrossRef]
  41. Fan, Y.; Chen, J.; Wang, Z.; Tan, T.; Li, S.; Li, J.; Wang, B.; Zhang, J.; Cheng, Y.; Wu, X.; et al. Soybean (Glycine max L. Merr.) seedlings response to shading: Leaf structure, photosynthesis and proteomic analysis. BMC Plant. Biol. 2019, 19, 464–471. [Google Scholar] [CrossRef]
  42. Jiang, C.D.; Wang, X.; Gao, H.Y.; Shi, L.; Chow, W.S. Systemic regulation of leaf anatomical structure, photosynthetic performance, and high-light tolerance in sorghum. Plant Physiol. 2011, 155, 1416–1424. [Google Scholar] [CrossRef]
  43. Driesen, E.; Van den Ende, W.; De Proft, M.; Saeys, W. Influence of environmental factors light, CO2, temperature, and relative humidity on stomatal opening and development: A review. Agronomy 2020, 10, 1975. [Google Scholar] [CrossRef]
  44. Raffo, A.; Mozzanini, E.; Ferrari Nicoli, S.; Lupotto, E.; Cervelli, C. Effect of light intensity and water availability on plant growth, essential oil production and composition in Rosmarinus officinalis L. Eur. Food Res. Technol. 2020, 246, 167–177. [Google Scholar] [CrossRef]
  45. Ghasemzadeh, A.; Nasiri, A.; Jaafar, H.Z.; Baghdadi, A.; Ahmad, I. Changes in phytochemical synthesis, chalcone synthase activity and pharmaceutical qualities of sabah snake grass (Clinacanthus nutans L.) in relation to plant age. Molecules 2014, 19, 17632–17648. [Google Scholar] [CrossRef] [PubMed]
  46. van der Sluis, A.A.; Dekker, M.; de Jager, A.; Jongen, W.M. Activity and concentration of polyphenolic antioxidants in apple: Effect of cultivar, harvest year, and storage conditions. J. Agric. Food Chem. 2001, 49, 3606–3613. [Google Scholar] [CrossRef] [PubMed]
  47. Awin, T.; Mediani, A.; Maulidiani Leong, S.W.; Faudzi, S.M.M.; Shaari, K.; Abas, F. Phytochemical and bioactivity alterations of Curcuma species harvested at different growth stages by NMR-based metabolomics. J. Food Compos. Anal. 2019, 77, 66–76. [Google Scholar] [CrossRef]
  48. Beatrice, P.; Saviano, G.; Reguzzoni, M.; Divino, F.; Fantasma, F.; Chiatante, D.; Montagnoli, A. Light spectra of biophilic LED-sourced system modify essential oils composition and plant morphology of Mentha piperita L. and Ocimum basilicum L. Front. Plant Sci. 2023, 14, 1093883. [Google Scholar] [CrossRef]
  49. Thakur, M.; Bhatt, V.; Kumar, R. Effect of shade level and mulch type on growth, yield and essential oil composition of damask rose (Rosa damascena Mill.) under mid hill conditions of Western Himalayas. PLoS ONE 2019, 14, e0214672. [Google Scholar] [CrossRef]
  50. Navehsi, F.M.; Abdossi, F.; Abbaszadeh, B.; Azimi, R.; Dianat, M. Effect of gamma rays on the essential oil and biochemical characteristics of the Satureja mutica Fisch & CA Mey. Sci. Rep. 2024, 14, 7581. [Google Scholar]
  51. Rumpf, J.; Burger, R.; Schulze, M. Statistical evaluation of DPPH, ABTS, FRAP, and Folin-Ciocalteu assays to assess the antioxidant capacity of lignins. Int. J. Biol. Macromol. 2023, 233, 123470. [Google Scholar] [CrossRef]
  52. Qaderi, M.M.; Martel, A.B.; Strugnell, C.A. Environmental Factors Regulate Plant Secondary Metabolites. Plants 2023, 12, 447. [Google Scholar] [CrossRef]
  53. Akarchariya, N.; Sirilun, S.; Julsrigival, J.; Chansakaowa, S. Chemical profiling and antimicrobial activity of essential oil from Curcuma aeruginosa Roxb., Curcuma glans K. Larsen & J. Mood and Curcuma cf. xanthorrhiza Roxb. collected in Thailand. Asian Pasific J. Trop. Med. 2017, 7, 881–885. [Google Scholar] [CrossRef]
  54. Stewart, P.; Boonsiri, P.; Puthong, S.; Rojpibulstit, P. Antioxidant activity and ultrastructural changes in gastric cancer cell lines induced by Northeastern Thai edible folk plant extracts. BMC Complement. Altern. Med. 2013, 13, 60. [Google Scholar] [CrossRef] [PubMed]
  55. Huang, Y.; Zhu, X.; Zhu, Y.; Wang, Z. Pinus koraiensis polyphenols: Structural identification, in vitro antioxidant activity, immune function and inhibition of cancer cell proliferation. Food Funct. 2021, 12, 4176–4198. [Google Scholar] [CrossRef] [PubMed]
  56. Iman, M.; Taheri, M.; Bahari, Z. The anti-cancer properties of neem (Azadirachta indica) through its antioxidant activity in the liver: Its pharmaceutics and toxic dosage forms. A literature review. J. Complement. Integr. Med. 2022, 19, 203–211. [Google Scholar] [CrossRef] [PubMed]
  57. Montenegro, J.; Dos Santos, L.S.; de Souza, R.G.G.; Lima, L.G.B.; Mattos, D.S.; Viana, B.P.P.B.; da Fonseca Bastos, A.C.S.; Muzzi, L.; Conte-Júnior, C.A.; Freitas-Silva, O.; et al. Bioactive compounds, antioxidant activity and antiproliferative effects in prostate cancer cells of green and roasted coffee extracts obtained by microwave-assisted extraction (MAE). Food Res. Int. Ott. Ont. 2021, 140, 110014. [Google Scholar] [CrossRef]
  58. Kuruburu, M.G.; Bovilla, V.R.; Leihang, Z.; Madhunapantula, S.V. Phytochemical-rich Fractions from Foxtail Millet (Setaria italica (L.) P. Beauv) Seeds Exhibited Antioxidant Activity and Reduced the Viability of Breast Cancer Cells In Vitro by Inducing DNA Fragmentation and Promoting Cell Cycle Arrest. Anticancer Agents Med. Chem. 2022, 22, 2477–2493. [Google Scholar] [CrossRef]
  59. Fitria, R.; Seno, D.S.H.; Priosoeryanto, B.P.; Hartanti; Nurcholis, W. Volatile compound profiles and cytotoxicity in essential oils from rhizome of Curcuma aeruginosa and Curcuma zanthorrhiza. Biodiversitas 2019, 20, 2943–2948. [Google Scholar] [CrossRef]
  60. Eddin, L.B.; Jha, N.K.; Goyal, S.N.; Agrawal, Y.O.; Subramanya, S.B.; Bastaki, S.M.A.; Ojha, S. Health benefits, pharmacological effects, molecular mechanisms, and therapeutic potential of α-bisabolol. Nutrients 2022, 14, 1370. [Google Scholar] [CrossRef]
  61. Ramazani, E.; Akaberi, M.; Emami, S.A.; Tayarani-Najaran, Z. Pharmacological and biological effects of alpha-bisabolol: An updated review of the molecular mechanisms. Life Sci. 2022, 304, 120728. [Google Scholar] [CrossRef]
  62. Cheng, B.; Hong, X.; Wang, L.; Cao, Y.; Qin, D.; Zhou, H.; Gao, D. Curzerene suppresses progression of human glioblastoma through inhibition of glutathione S-transferase A4. CNS Neurosci. Ther. 2022, 28, 690–702. [Google Scholar] [CrossRef]
  63. Wang, Y.; Li, J.; Guo, J.; Wang, Q.; Zhu, S.; Gao, S.; Yang, C.; Wei, M.; Pan, X.; Zhu, W.; et al. Cytotoxic and Antitumor Effects of Curzerene from Curcuma longa. Planta Med. 2017, 83, 23–29. [Google Scholar] [CrossRef]
  64. Rodrigo, G.; Almanza, G.R.; Cheng, Y.; Peng, J.; Hamann, M.; Duan, R.D.; Åkesson, B. Antiproliferative effects of curcuphenol, a sesquiterpene phenol. Fitoterapia 2010, 81, 762–766. [Google Scholar] [CrossRef] [PubMed]
  65. Figueiredo, P.L.B.; Pinto, L.C.; da Costa, J.S.; da Silva, A.R.C.; Mourão, R.H.V.; Montenegro, R.C.; da Silva, J.K.R.; Maia, J.G.S. Composition, antioxidant capacity and cytotoxic activity of Eugenia uniflora L. chemotype-oils from the Amazon. J. Ethnopharmacol. 2019, 232, 30–38. [Google Scholar] [CrossRef] [PubMed]
  66. Russell, K.; Jacob, S.E. Bisabolol. Dermat. Contact Atopic. Occup. Drug. 2010, 21, 57–58. [Google Scholar] [CrossRef]
  67. Egbuta, M.A.; McIntosh, S.; Waters, D.L.; Vancov, T.; Liu, L. In vitro anti-inflammatory activity of essential oil and β-bisabolol derived from cotton gin trash. Molecules 2022, 27, 526. [Google Scholar] [CrossRef] [PubMed]
  68. Abutaha, N.; Nasr, F.A.; Al-Zharani, M.; Alqahtani, A.S.; Noman, O.M.; Mubarak, M.; Abdelhabib, S.; Wadaan, M.A. Effects of Hexane Root Extract of Ferula hermonis Boiss. on Human Breast and Colon Cancer Cells: An In Vitro and In Vivo Study. BioMed. Res. Int. 2019, 2019, 3079895. [Google Scholar] [CrossRef]
  69. Biltekin, S.N.; Karadağ, A.E.; Demirci, F.; Demirci, B. In Vitro Anti-Inflammatory and Anticancer Evaluation of Mentha spicata L. and Matricaria chamomilla L. Essential Oils. ACS Omega 2023, 8, 17143–17150. [Google Scholar] [CrossRef]
  70. Belayachi, L.; Aceves-Luquero, C.; Merghoub, N.; de Mattos, S.F.; Amzazi, S.; Villalonga, P.; Bakri, Y. Induction of cell cycle arrest and apoptosis by ormenis eriolepis a morrocan endemic plant in various human cancer cell lines. Afr. J. Tradit. Complement. Altern. Med. 2017, 14, 356–373. [Google Scholar] [CrossRef]
  71. Scalvenzi, L.; Grandini, A.; Spagnoletti, A.; Tacchini, M.; Neill, D.; Ballesteros, J.L.; Sacchetti, G.; Guerrini, A. Myrcia splendens (Sw.) DC. (syn. M. fallax (Rich.) DC.) (Myrtaceae) essential oil from Amazonian Ecuador: A chemical characterization and bioactivity profile. Molecules 2017, 22, 1163. [Google Scholar]
  72. Taglialatela-Scafati, O.; Pollastro, F.; Cicione, L.; Chianese, G.; Bellido, M.L.; Munoz, E.; Özen, H.C.; Toker, Z.; Appendino, G. STAT-3 Inhibitory Bisabolanes from Carthamus glaucus. J. Nat. Prod. 2012, 75, 453–458. [Google Scholar] [CrossRef]
  73. Soltanian, S.; Mohamadi, N.; Rajaei, P.; Khodami, M.; Mohammadi, M. Phytochemical composition, and cytotoxic, antioxidant, and antibacterial activity of the essential oil and methanol extract of Semenovia suffruticosa. Avicenna J. Phytomed. 2019, 9, 143–152. [Google Scholar]
  74. Nohara, L.L.; Ellis, S.L.S.; Dreier, C.; Dada, S.; Saranchova, I.; Choi, K.B.; Munro, L.; Al Haddad, E.; Pfeifer, C.G.; Coyle, K.M.; et al. A novel cell-based screen identifies chemical entities that reverse the immune-escape phenotype of metastatic tumours. Front. Pharmacol. 2023, 14, 1119607. [Google Scholar] [CrossRef] [PubMed]
  75. Ellis, S.L.; Dada, S.; Nohara, L.L.; Saranchova, I.; Munro, L.; Pfeifer, C.G.; Eyford, B.A.; Morova, T.; Williams, D.E.; Cheng, P.; et al. Curcuphenol possesses an unusual histone deacetylase enhancing activity that counters immune escape in metastatic tumours. Front Pharmacol. 2023, 14, 1119620. [Google Scholar] [CrossRef] [PubMed]
  76. Abd-ElGawad, A.M.; El Gendy, A.E.N.G.; Assaeed, A.M.; Al-Rowaily, S.L.; Omer, E.A.; Dar, B.A.; Al-Taisan, W.A.; Elshamy, A.I. Essential Oil Enriched with Oxygenated Constituents from Invasive Plant Argemone ochroleuca Exhibited Potent Phytotoxic Effects. Plants 2020, 9, 998. [Google Scholar] [CrossRef] [PubMed]
  77. Grover, M.; Behl, T.; Virmani, T. Phytochemical Screening, Antioxidant Assay and Cytotoxic Profile for Different Extracts of Chrysopogon zizanioides Roots. Chem. Biodivers. 2021, 18, e2100012. [Google Scholar] [CrossRef]
  78. Ez-Zriouli, R.; ElYacoubi, H.; Imtara, H.; Mesfioui, A.; ElHessni, A.; Al Kamaly, O.; Zuhair Alshawwa, S.; Nasr, F.A.; Benziane Ouaritini, Z.; Rochdi, A. Chemical Composition, Antioxidant and Antibacterial Activities and Acute Toxicity of Cedrus atlantica, Chenopodium ambrosioides and Eucalyptus camaldulensis Essential Oils. Molecules 2023, 28, 2974. [Google Scholar] [CrossRef]
  79. Ogundajo, A.L.; Ewekeye, T.; Sharaibi, O.J.; Owolabi, M.S.; Dosoky, N.S.; Setzer, W.N. Antimicrobial Activities of Sesquiterpene-Rich Essential Oils of Two Medicinal Plants, Lannea egregia and Emilia sonchifolia, from Nigeria. Plants. 2021, 10, 488. [Google Scholar] [CrossRef]
  80. Sanad, H.; Belattmania, Z.; Nafis, A.; Hassouani, M.; Mazoir, N.; Reani, A.; Hassani, L.; Vasconcelos, V.; Sabour, B. Chemical Composition and In Vitro Antioxidant and Antimicrobial Activities of the Marine Cyanolichen Lichina pygmaea Volatile Compounds. Mar. Drugs 2022, 20, 169. [Google Scholar] [CrossRef]
  81. El Mihyaoui, A.; Esteves da Silva, J.C.G.; Charfi, S.; Candela Castillo, M.E.; Lamarti, A.; Arnao, M.B. Chamomile (Matricaria chamomilla L.): A Review of Ethnomedicinal Use, Phytochemistry and Pharmacological Uses. Life 2022, 12, 479. [Google Scholar] [CrossRef]
  82. Alahmady, N.F.; Alkhulaifi, F.M.; Momenah, M.A.; Alharbi, A.A.; Allohibi, A.; Alsubhi, N.H.; Alhazmi, W.A. Biochemical characterization of chamomile essential oil: Antioxidant, antibacterial, anticancer and neuroprotective activity and potential treatment for Alzheimer’s disease. Saudi J. Biol. Sci. 2024, 31, 103912. [Google Scholar] [CrossRef]
  83. Lin, J.; Dou, J.; Xu, J.; Aisa, H.A. Chemical composition, antimicrobial and antitumor activities of the essential oils and crude extracts of Euphorbia macrorrhiza. Molecules 2012, 17, 5030–5039. [Google Scholar] [CrossRef]
  84. Djihane, B.; Wafa, N.; Elkhamssa, S.; Maria, A.E.; Mihoub, Z.M. Chemical constituents of Helichrysum italicum (Roth) G. Don essential oil and their antimicrobial activity against Gram-positive and Gram-negative bacteria, filamentous fungi and Candida albicans. Saudi Pharm. J. 2017, 25, 780–787. [Google Scholar] [CrossRef] [PubMed]
  85. Zhang, C.; Xie, Y.; Qiu, W.; Mei, J.; Xie, J. Antibacterial and antibiofilm efficacy and mechanism of ginger (Zingiber officinale) essential oil against Shewanella putrefaciens. Plants 2023, 12, 1720. [Google Scholar] [CrossRef] [PubMed]
  86. Zhang, J.; Pan, Q.; Zhang, X.; Tana. Chemical Composition and Antioxidant and Antibacterial Potencies of the Artemisia ordosica Aerial Parts Essential Oil during the Vegetative Period. Molecules 2022, 27, 8898. [Google Scholar] [CrossRef] [PubMed]
  87. Pandini, J.A.; Pinto, F.G.S.; Scur, M.C.; Santana, C.B.; Costa, W.F.; Temponi, L.G. Chemical composition, antimicrobial and antioxidant potential of the essential oil of Guarea kunthiana A. Juss. Braz. J. Biol. 2017, 78, 53–60. [Google Scholar] [CrossRef]
  88. Wang, X.; Shen, Y.; Thakur, K.; Han, J.; Zhang, J.G.; Hu, F.; Wei, Z.J. Antibacterial activity and mechanism of ginger essential oil against Escherichia coli and Staphylococcus aureus. Molecules 2020, 25, 3955. [Google Scholar] [CrossRef]
Plants 13 02682 g001
Figure 1. (a) The workflow of essential oil extraction from C. xanthorrhiza Roxb. rhizomes. (b) Yield (% v/w) of C. xanthorrhiza essential oil. Extraction was performed in 3 independent plants, with statistical analysis by one-way ANOVA followed by Tukey’s post hoc test, *** p < 0.001. S0, no shade; S25, 25% shade; S50, 50% shade; S75, 75% shade.
Figure 1. (a) The workflow of essential oil extraction from C. xanthorrhiza Roxb. rhizomes. (b) Yield (% v/w) of C. xanthorrhiza essential oil. Extraction was performed in 3 independent plants, with statistical analysis by one-way ANOVA followed by Tukey’s post hoc test, *** p < 0.001. S0, no shade; S25, 25% shade; S50, 50% shade; S75, 75% shade.
Plants 13 02682 g001
Plants 13 02682 g002
Figure 2. Chemical compositions of C. xanthorrhiza Roxb. essential oil. (a) Pie chart displaying the groups of secondary metabolites in C. xanthorrhiza essential oil from 4 shading conditions detected by GC-MS. (b) Heatmap displaying the abundance of 64 compounds detected by GC-MS in C. xanthorrhiza essential oil from each shade condition. Three replicates of EO extraction were pooled and subjected to chemical compound analysis.
Figure 2. Chemical compositions of C. xanthorrhiza Roxb. essential oil. (a) Pie chart displaying the groups of secondary metabolites in C. xanthorrhiza essential oil from 4 shading conditions detected by GC-MS. (b) Heatmap displaying the abundance of 64 compounds detected by GC-MS in C. xanthorrhiza essential oil from each shade condition. Three replicates of EO extraction were pooled and subjected to chemical compound analysis.
Plants 13 02682 g002
Plants 13 02682 g003
Figure 3. Antioxidant activity of C. xanthorrhiza Roxb. essential oil measured by DPPH and FRAP assays. n = 3, statistical analysis by one-way ANOVA followed by Tukey’s post hoc test, * p < 0.05, ** p < 0.01, *** p < 0.001. S0, no shade; S25, 25% shade; S50, 50% shade; S75, 75% shade. DPPH, 2,2′-diphenyl-1-picrylhydrazyl; DW, dry weight; FRAP, ferric reducing antioxidant power; TE, Trolox equivalent; DW: dry weight.
Figure 3. Antioxidant activity of C. xanthorrhiza Roxb. essential oil measured by DPPH and FRAP assays. n = 3, statistical analysis by one-way ANOVA followed by Tukey’s post hoc test, * p < 0.05, ** p < 0.01, *** p < 0.001. S0, no shade; S25, 25% shade; S50, 50% shade; S75, 75% shade. DPPH, 2,2′-diphenyl-1-picrylhydrazyl; DW, dry weight; FRAP, ferric reducing antioxidant power; TE, Trolox equivalent; DW: dry weight.
Plants 13 02682 g003
Plants 13 02682 g004
Figure 4. Antiproliferative activity of C. xanthorrhiza Roxb. essential oil (4 µg/mL) measured in vero (left), MCF-7 (middle), or HeLa (right) cells. n = 3, statistical analysis by one-way ANOVA followed by Tukey’s post hoc test, ** p < 0.01, *** p < 0.001. S0, no shade; S25, 25% shade; S50, 50% shade; S75, 75% shade. Doxo: doxorubicin (0.2 µg/mL).
Figure 4. Antiproliferative activity of C. xanthorrhiza Roxb. essential oil (4 µg/mL) measured in vero (left), MCF-7 (middle), or HeLa (right) cells. n = 3, statistical analysis by one-way ANOVA followed by Tukey’s post hoc test, ** p < 0.01, *** p < 0.001. S0, no shade; S25, 25% shade; S50, 50% shade; S75, 75% shade. Doxo: doxorubicin (0.2 µg/mL).
Plants 13 02682 g004
Plants 13 02682 g005
Figure 5. Antibacterial activity of C. xanthorrhiza Roxb. essential oil (4 µg/mL) tested against S. aureus (left) or E. coli (right). n = 3, statistical analysis by one-way ANOVA followed by Tukey’s post hoc test, */# p < 0.05, ### p < 0.001. * Analysis was performed by using S0 as reference. # Analysis was performed by using KS as reference. S0, no shade; S25, 25% shade; S50, 50% shade; S75, 75% shade. KS: kanamycin sulfate (1 mg/mL).
Figure 5. Antibacterial activity of C. xanthorrhiza Roxb. essential oil (4 µg/mL) tested against S. aureus (left) or E. coli (right). n = 3, statistical analysis by one-way ANOVA followed by Tukey’s post hoc test, */# p < 0.05, ### p < 0.001. * Analysis was performed by using S0 as reference. # Analysis was performed by using KS as reference. S0, no shade; S25, 25% shade; S50, 50% shade; S75, 75% shade. KS: kanamycin sulfate (1 mg/mL).
Plants 13 02682 g005
Table 1. Volatile compounds identified from the essential oils of various shades of C. xanthorrhiza. Roxb. Three replicates of EO extraction were pooled and subjected to chemical compound analysis.
Table 1. Volatile compounds identified from the essential oils of various shades of C. xanthorrhiza. Roxb. Three replicates of EO extraction were pooled and subjected to chemical compound analysis.
No.CompoundsGroup CompoundsMFMW (g/mol)RT%Area
N0%N25%N50%N75%N0%N25%N50%N75%
1EucalyptolMonoterpenesC10H18O15412.56612.56212.56012.5510.690.881.220.60
2Camphene hydrateMonoterpenesC10H18O15416.71416.70616.70016.6890.991.181.190.79
3β-elemeneSesquiterpenesC15H2420423.48623.43323.44223.4330.490.470.430.36
4α-CurcumeneSesquiterpenesC15H2220225.91825.775-25.7677.407.29-5.81
5KhuseneSesquiterpenesC15H2420426.31726.30826.30826.3000.260.270.280.23
6α-CedreneSesquiterpenesC15H2420426.65626.62526.48326.6038.116.796.444.32
7EpicubebolSesquiterpenesC15H26O222-26.74526.66726.736-0.410.360.28
8Dys-oxylonenSesquiterpenesC15H24204-26.817-26.800-0.17-0.13
97-epi-cis-sesqui-sabinene hydrateSesquiterpenesC15H26O222-26.875-26.865-0.43-0.37
10Elemene isomerSesquiterpenesC15H24204-26.982-26.974-0.27-0.18
11α-calameneSesquiterpenesC15H22202-27.41727.392 -0.200.17
12γ-eudesmolSesquiterpenesC15H260222--29.50730.043--0.611.34
13λ-cadineneSesquiterpenesC15H2420426.833---0.16---
14β-sesqui-sabineneSesquiterpenesC15H2420426.890-26.87226.8720.42-0.550.55
15ElemolSesquiterpenesC15H26O22227.48827.47927.47627.4810.550.310.290.35
16LinaloolMonoterpenesC10H18015427.742-27.733-0.18-0.19-
17β-germacreneSesquiterpenesC15H2420427.82627.81327.81027.8101.681.651.391.28
18Epi-curzerenoneSesquiterpenesC15H18O223028.82128.81328.80028.8547.727.897.458.32
19α-terpineneDiterpenoidC20H3227228.971-28.95628.9780.74-0.590.94
20Artemisia ketoneMonoterpenesC10H16O15229.04229.04229.042-0.250.280.26-
21Germacrene D-4-olSesquiterpenesC15H26O22229.26729.25829.25829.2630.200.190.270.63
22Iso-spathulenolSesquiterpenesC15H24O22029.40829.41129.40229.4160.720.680.701.07
23α-acorenolSesquiterpenesC15H26022229.55729.553--0.580.73--
241-Heptatria-cotanolFatty alcoholsC37H76O53629.632---0.20---
25AtractylonSesquiterpenesC15H20O21630.15830.15830.150-0.280.300.25-
26GermacroneSesquiterpenesC15H22021831.00030.98830.97831.0156.546.246.136.62
27CembreneDiterpenoidC20H3227231.55331.53331.53226.4520.800.670.640.42
28XantorrhizolSesquiterpenesC15H22O21832.30632.28932.28832.2835.803.054.666.03
29Naphto [2,3-b] furan-4 (6H)-oneCarboxylic esterC15H180223032.41432.39832.39732.4350.600.580.410.67
30(Z)-betha-curcumen-12-olSesquiterpenesC15H24O22032.46729.96229.16329.1750.090.750.641.05
31α-CopaeneSesquiterpenesC15H26O2238--32.742---0.30-
32(E)-Iso-valencenalSesquiterpenesC15H22O21832.628---0.95---
33(4-Isopropyl-1-methyl-6,7-dimethylenebicyclo [3.2.1]oct-8-yl)methanolSesquiterpenesC15H24O220-32.614---0.81--
34Cedren-13-ol, 8-SesquiterpenesC15H24O22032.71832.704--0.320.30--
35(1R,7S,E)-7-Isopropyl-4,10-dimethylenecyclodec-5-enolSesquiterpenesC15H24O220--32.16032.644--0.911.21
36SqualeneTriterpenesC30H5041032.833-32.833-0.07-0.07-
37Caryo-phylleneSesquiterpenesC15H24204---24.299---0.31
38ElixeneSesquiterpenesC15H24204---24.525---0.62
39CurzereneSesquiterpenesC15H20O216---26.125---3.07
40Sesqui-cineoleSesquiterpenesC15H26O222---26.676---0.11
41trans-Sesqui-sabinene hydrateSesquiterpenesC15H26O222---27.351---0.63
42(Z)-betha-sesqui-sabinene-hydrateSesquiterpenesC15H26O222---27.631---1.54
43γ-himachaleneSesquiterpenesC15H24204---28.501---1.56
4410-epi-Y-EudesmolSesquiterpenesC15H26O222---29.483---0.41
45EpiglobulolSesquiterpenesC15H26O222---29.557---0.53
461,8-Nonadiene, 2,7-di-methyl-5-(1-methyl-ethenyl)-MonoterpenoidsC14H24192---28.632---0.42
471,10-epi-cubenolSesquiterpenesC15H26O222---29.750---0.44
48Guaiol acetateSesquiterpenesC17H28O2264---29.833---0.44
49Neo-intermedeolSesquiterpenesC15H26O222---30.117---0.60
50BulnesolSesquiterpenesC15H26O222---30.258---0.28
51β-BisabololSesquiterpenesC15H26O222---30.368---2.18
521,1,4,7-Tetramethyldecahydro-1H-cyclopropa[e]azulene-4,7-diolSesquiterpenesC15H26O2238---30.531---0.61
53α-bisabololSesquiterpenesC15H26O222---30.739---0.79
54Menthyl anthranilateMonoterpenoidsC17H25NO2275---30.833---0.76
55Curcu-phenolSesquiterpenesC15H22O218---31.272---1.83
56β-elemeneMonoterpenoidsC14H24192---32.864---0.37
5724-Norursa-3,12-dien-11-oneSesquiterpenesC29H44O408---32.087---1.86
58α-copaen-11-olSesquiterpenesC15H24O220---33.202---0.10
59Dihydro-piperlonguminineAmidesC16H21NO3275---33.983---0.07
60Dehydro-fukinoneSesquiterpenesC15H22O218---35.143---0.03
611,2-Hexadecane-diolFatty AlcoholsC16H34O2258---36.334---0.17
62Hexa-decamethyl-cyclo-octasiloxaneOrganosiloxaneC16H48O8Si8592---38.479---0.08
63Dihexadecyl phosphateDialkyl phosphateC32H67O4P546---47.614---0.09
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content.

Share and Cite

MDPI and ACS Style

Nurcholis, W.; Rahmadansah, R.; Astuti, P.; Priosoeryanto, B.P.; Arianti, R.; Kristóf, E. Comparative Analysis of Volatile Compounds and Biochemical Activity of Curcuma xanthorrhiza Roxb. Essential Oil Extracted from Distinct Shaded Plants. Plants 2024, 13, 2682. https://doi.org/10.3390/plants13192682

AMA Style

Nurcholis W, Rahmadansah R, Astuti P, Priosoeryanto BP, Arianti R, Kristóf E. Comparative Analysis of Volatile Compounds and Biochemical Activity of Curcuma xanthorrhiza Roxb. Essential Oil Extracted from Distinct Shaded Plants. Plants. 2024; 13(19):2682. https://doi.org/10.3390/plants13192682

Chicago/Turabian Style

Nurcholis, Waras, Rahmadansah Rahmadansah, Puji Astuti, Bambang Pontjo Priosoeryanto, Rini Arianti, and Endre Kristóf. 2024. "Comparative Analysis of Volatile Compounds and Biochemical Activity of Curcuma xanthorrhiza Roxb. Essential Oil Extracted from Distinct Shaded Plants" Plants 13, no. 19: 2682. https://doi.org/10.3390/plants13192682

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop