Next Article in Journal
Bioponics—An Organic Closed-Loop Soilless Cultivation System: Yields and Characteristics Compared to Hydroponics and Soil Cultivation
Next Article in Special Issue
Changes in Drip Irrigation Water Distribution Patterns Improve Fruit Quality and Economic Water Productivity in Early-Season Lemon Trees
Previous Article in Journal
Resistance to Anthracnose Rot Disease in Capsicum
Previous Article in Special Issue
Local Agro-Environmental Conditions Impact Fruit Quality, Sensory Properties and Consumer Acceptance of Long Shelf-Life Tomatoes
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Agronomy
Volume 13
Issue 6
10.3390/agronomy13061435
agronomy-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

Relationship between Phenotypes and Chemical Profiling of Boesenbergia rotunda Collected from Different Habitats of Northern Thailand

by
Sureerat Thomya
1,
Malaiporn Wongkaew
2,3,
Weenun Bundithya
4,
Chompunut Lumsangkul
5,6,
Pensak Jantrawut
7,8,9 and
Sarana Rose Sommano
2,8,9,*
1
Interdisciplinary Program in Biotechnology, Graduate School, Chiang Mai University, Chiang Mai 50200, Thailand
2
Plant Bioactive Compound Laboratory, Faculty of Agriculture, Chiang Mai University, Chiang Mai 50200, Thailand
3
Program in Food Production and Innovation, College of Integrated Science and Technology, Rajamangala University of Technology Lanna, Chiang Mai 50220, Thailand
4
Department of Plant and Soil Science, Faculty of Agriculture, Chiang Mai University, Chiang Mai 50200, Thailand
5
Department of Animal and Aquatic Sciences, Faculty of Agriculture, Chiang Mai University, Chiang Mai 50200, Thailand
6
Multidisciplinary Research Institute, Chiang Mai University, Chiang Mai 50200, Thailand
7
Department of Pharmaceutical Sciences, Faculty of Pharmacy, Chiang Mai University, Chiang Mai 50200, Thailand
8
Cluster of Valorization and Bio-Green Transformation for Translational Research Innovation of Raw Materials and Products, Chiang Mai University, Chiang Mai 50200, Thailand
9
Cluster of Agro Bio-Circular-Green Industry (Agro BCG), Chiang Mai University, Chiang Mai 50200, Thailand
*
Author to whom correspondence should be addressed.
Agronomy 2023, 13(6), 1435; https://doi.org/10.3390/agronomy13061435
Submission received: 14 April 2023 / Revised: 9 May 2023 / Accepted: 20 May 2023 / Published: 23 May 2023
(This article belongs to the Special Issue Effects of Agronomical Practices on Crop Quality and Sensory Profile)
Browse Figures
Versions Notes

Abstract

:
Fingerroot [Boesenbergia rotunda (Linn.) Mansf] is known to consist of compounds such as panduratin A and pinostrobin with inhibitory property against SARS-CoV-2 infection at both the pre-entry and post-infection phases. Consequently, demand for good phenotype selection has increased in the post-pandemic era. However, the availability of important active ingredients may be influenced by genetic variables and agronomic aspects. Thus, the purpose of this study was to investigate the relationship between characteristics and genotype, and to preliminarily compare the phytochemical profiles of the landraces collected in Northern Thailand. Five local landraces and two wild types were gathered for ex situ collection for morphological assessments, genetic evaluation, and bioactive ingredients (mainly antioxidative potentials and amounts of the active flavonoids). The morphological data were able to distinguish the plant samples to those of wild, cultivated, and adaptive types, which was confirmed by their distinctive genetic variations. However, there was no correlation between the physical attributes and the amount of their bioactive constituents. It was also observed that the adaptation of plants to environmental conditions had a pronounced impact on secondary metabolite biosynthesis, and that such adaptations were likely influenced by genetic differentiation. The findings from this study could potentially be used to improve the cultivation, selection, and breeding of this plant species for desired traits such as increased bioactive compound content, or for conservation and restoration efforts of the landraces and wild types.

1. Introduction

The genus Bosenbergia has a wide distribution across tropical regions and is known for its use in spices and herbal medicine in Southeast Asia, India, Sri Lanka, and Southern China [1,2,3]. There are over 80 recognized species within this genus, and Thailand is considered to be one of the richest habitats with over 20 species present [3,4]. Among those, fingerroot (Bosenbergia rotunda), or locally known as Krachai kaw in Thai, has recently attracted attention for its potential use as a preventative medicine [1,2,3]. Like its siblings in the Zingiberacea family, Krachai kaw is a small herbaceous plant with short, fleshy, or slender rhizomes in the shape of fingers growing from the mother rhizomes [2,4,5]. The rhizomes are the nutrient accumulation parts, and therefore are the rich source of active ingredients, comprising various aromatic compounds [2,6]. Propagation of B. rotunda can be done vegetatively; however, the hybridization could also happen naturally by cross-pollination or a breeding program, which provide variations in genetic diversity [7,8]. Although the species richness of B. rotunda in northern Thailand is diverse, there is limited documentation regarding the diverse genetic pools and their relation to the morphological characteristics. The phytoconstituents from the rhizomes of B. rotunda are generally classified into two major groups: flavanones (e.g., alpinetin, pinostrobin, and pinocembrin) and chalcones (e.g., boesenbergin, cardamonin, panduratin A, and 4-hydroxypanduratin A) [9]. They contribute vastly to many different pharmaceutical activities, including antibacterial, anti-allergic, anti-tumor, anti-mutation, anti-fungal, anti-anxiety, and anti-inflammatories [10,11,12,13,14]. More importantly, the recent report advised that panduratin A had a potential inhibition activity against SARS-CoV-2 (COVID-19) and prevented viral replication at both the pre-entry and post-infection phases, which is more potent than the hydroxychloroquine drug used for COVID-19 treatment [7]. Meanwhile, pinostrobin can inhibit intestinal smooth muscle contractions and pose antiviral, antioxidant, anti-leukaemic, anti-inflammatory, anti-aromatase, anti-microbial, anti-ulcer, and anti-tumor properties [15,16]. Nonetheless, the contents of these bioactive compounds in B. rotunda collected from various cultivation areas are variable, and the information on the phenotypic relationship with the chemical profiling is also lacking.
The demand for high-quality B. rotunda has been increasing in both the food and pharmaceutical industries, especially in the post-COVID era where there is a growing interest in natural remedies. This trend has led to a need for selecting good phenotypes for commercial cultivation. It is essential to evaluate the morphological traits of the plant to determine the level of bioactive compounds present in it, which could be useful for developing a plant breeding program. From this rationale, the aims of this study were to investigate the morphological differences among locally collected plant accessions and study their relationship with their phytochemical profiling. By identifying the traits that correlate with high bioactive compound levels, plant breeders can select good phenotypes with desirable characteristics and breed new cultivars with increased concentrations of such active compounds. This can ultimately lead to the production of commercial B. rotunda type with improved medicinal and nutritional properties, meeting the increasing demand for high-quality plants in the market.

2. Materials and Methods

2.1. Reagent and Chemicals

The genomic DNA Extraction Kit for plant material was from RBC Bioscience (New Taipei City, Taipei). Polymorphic DNA primer sequences were supplied by Integrated DNA Technologies (Coralville, IA, USA). Taq DNA polymerase 2x Master Mix RED and High Range DNA Ladder were purchased from Ampliqon PCR Enzymes & Reagents (Stenhuggervej, Odense M, Denmark). Novel Juice (DNA Staining Reagent) was supplied by Bio-Helix (New Taipei City, Taiwan). Standard pinostrobin and panduratin A were obtained from Chem Faces (Wuhan, Hubei). Gallic acid monohydrate, 2,2′-azino-bis (3-ethylbenzthiazoline-6-sulphonic acid) diammonium salt, (+)—Catechin hydrate (sum of enantiomers, HPLC), and 2,2-Diphenyl-1-picrylhydrazyl were from Sigma-Aldrich (St. Louis, MO, USA). Folin–Ciocalteu’s phenol reagent was supplied by Merckmillipore (Billerica, MA, USA). Sodium hydrogen carbonate, sodium nitrile, aluminium chloride (Hexahydrate), sodium hydroxide (Micropearls), Potassium dihydrogen orthophosphate anhydrous, potassium persulphate, acetonitrile and glacial acetic acid (HPLC grade solvents). and Formic acid and Methanol (LCMS grade solvents) were purchased from RCI Labscan Limited, Bangkok, Thailand.

2.2. Plant Materials

Seedling (rhizomes) of the B. rotunda were collected from their natural habitats and cultivation areas in Chiang Mai, Lamphun, and Lampang. They were five landraces and two wild types as described by the ending letters C and W in Table 1. At the Faculty of Agriculture, Chiang Mai University, Chiang Mai (CMU), Thailand, the rhizomes were cultivated in 8 × 13 inch pots with a substrate comprising of loam soil and animal manure in a 2:1 ratio. For each line, 20 seedling were cloned and were arranged randomly in four blocks and each block contained five replications. They were maintained in a greenhouse under 50% shading with daily irrigation and 200 g/plant of manual organic fertilizer (1:20 farmyard manure-to-humus ratio) application at 65 days after cultivation.

2.3. Phenotypic Characterization

The morphological data were recorded for the above- and underground parts of B. rotunda Plant growths were measured once a week after planting and continuing until 17 weeks (before the plant entered its dormancy period), by recording leaf width (cm), leaf length (cm), stem width (cm), and stem height (cm). The rhizomes were harvested at 18 weeks after cultivation when the aboveground organs turned yellow and collapsed, when the rhizomes have reached their maturity [17]. The rhizomes were carefully washed and weighed (g) before and after the fingers were separated from the mother rhizomes. For the rhizomes, width (cm) and length (cm) were also measured. Afterward, these underground parts were freeze-dried and thereafter ground to powder for further analysis of active ingredients.

2.4. Genetic Variation Analysis of B. rotunda

2.4.1. DNA Extraction

Total genomic DNA was extracted using the technique outlined in the manual for the genomic DNA extraction kit. The 50 mg of the freeze-dried powder samples were pulverized using liquid nitrogen in an Eppendorf tube, with step-wise additions of the buffers provided in the kit. The purified DNA was obtained by centrifuging at 20,627× g for 30 s. The quality and quantity of DNA were then analyzed using a Nano Drop Spectrophotometers, Thermo Scientific/NanoDrop 2000C (AG-BIO/PERDO-CHE, Bangkok, Thailand). The DNA samples were stored at −20 °C prior to random amplified polymorphic DNA (RAPD) analysis.

2.4.2. RAPD-PCR

RAPD-PCR was performed in accordance with the Ampliqon PCR enzyme & Re-agents manual’s instructions. For RAPD analysis of the genomic DNA, 10 primers were chosen [18,19] (Table S1) and PCR reactions were operated following the procedure described in Supplementary Table S2. All reactions were carried out with a K960 Thermal cycler (Heal Force) using the following program: 1 cycle, 95 °C, 3 min; 40 cycles, 95 °C, 30 s; 37 °C, 1 min; 72 °C, 30 s; 1 cycle, 72 °C, 5 min. Consequently, the sample was separated in a 1% agarose gel in 1× TBE buffer. The PCR products were stained with Novel juice DNA staining reagent and run at 135 V for 22 min. The gels were accordingly visualized using the LED transilluminator, TT-BLT-470; Hercuvan (Cambridge, CB24 6WZ, UK).

2.5. Phytochemical Analyses

2.5.1. Rhizome Extraction of B. rotunda

Three grams of B. rotunda powder was dissolved in 30 mL of 80% (v/v) ethanol and macerated for 24 h at room temperature before being separated with Whatman No. 1 filter paper as described by Sangta et al. [20]. The residue was extracted three times, and the filtrates were combined. The solution was evaporated to dryness in the rotary evaporator resulting in the crude ethanol extract.

2.5.2. Total Phenolic Content

The crude extract (30 µL) was combined with 60 µL of Folin–Ciocalteu reagent and neutralized with 210 µL of 6% w/v NaCO3 solution, which was added, mixed, and left in the dark for 90 min as described by Sangta et al. [20]. A UV-Vis spectrophotometer was used to measure the absorbance at a wavelength of 725 nm (SPECTROstar, BMG LABTECH, Offenburg, Germany). Gallic acid calibration curve was constructed at different concentrations of gallic acid (0–1000 mg/mL). The total phenolic content was evaluated in milligrams of gallic acid equivalents per dried sample.

2.5.3. Total Flavonoid Content

The total flavonoid content was determined using a modified Sunanta et al. [21] technique. The ethanol extract (25 µL) was reacted with 125 µL of distilled water and 7.5 µL of a 5% NaNO2 solution. Before adding 15 µL of 10% AlCl3·6H2O solution, the mixture was allowed to react at room temperature for 5 min. After 6 min of incubation time, 50 µL of 1 M NaOH solution and 27.5 µL of distilled water were added. The UV-Vis spectrophotometer was used to measure the absorbance at 510 nm (SPECTROstar, BMG LABTECH, Offenburg, Germany). The catechin calibration curve was developed at concentrations of 0 to 1000 mg/mL. The total flavonoid content was reported in milligrams of catechin equivalents per dried sample.

2.5.4. Antioxidant Potential

DPPH Radical Scavenging Activity

The DPPH free radical scavenging activity was operated in accordance with Sangta, Wongkaew, Tangpao, Withee, Haituk, Arjin, Sringarm, Hongsibsong, Sutan and Pusadee [20]. A 25 µL of the ethanol extracts was incubated with 250 µL of 0.2 mM DPPH (2,2-diphenyl-1-picrylhydrazyl) for 30 min in the dark at room temperature. The absorbance was measured at 517 nm using a UV-Vis spectrophotometer (SPECTROstar, BMG LABTECH, Offenburg, Germany). DPPH radical scavenging activity was calculated using the following equation;
DPPH scavenging activity (%) = [(A control − A sample)/A control] × 100
where A control is a DPPH absorbance and A sample is the sample absorbance.

ABTS•+ Radical Cation Activity

The ABTS [2,2-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid)] assay was determined using the method described by Sangta et al. [20]. The working solution was prepared by combining the two stock solutions, 7.0 mM ABTS solution and 2.45 mM K2S2O8 solution. The solutions were mixed in equal amounts and left to react in the dark at room temperature for 24 h. The ABTS solution was arranged by diluting 1.0 mL of ABTS with 60 mL of 80% ethanol to achieve an absorbance of 0.70 ± 0.02 units at 734 nm. A 10 µL of the ethanol fingerroot extracts and 200 µL of ABTS working solution were added in each well, shaken and incubated for 30 min at room temperature. The absorbance was accordingly measured at 734 nm. The ABTS scavenging activity was estimated using the following equation;
ABTS radical scavenging activity (%) = [(A control − A sample)]/(A control)] × 100
where A control is the absorbance of ABTS radical mixed with 80% methanol; A sample is the absorbance of ABTS radical reacted with sample extract/standard.

2.5.5. Analyses of Pinostrobin and Panduratin A by High-Performance Liquid Chromatography (HPLC)

The crude extract was dissolved in 95% methanol to a final concentration of 0.1 mg/mL and analyzed for the pinostrobin and panduratin A using the HPLC (Shimadzu, Kyoto, Japan) with an automatic injection (SIL-20ACHT), diode array detection (CTO-20AC), pump (LC-20AD), and automatic control. Ultra-Aqueous C18 (250 4.6 mm, 5 m) was utilized for reverse-phase column chromatography (RESTEK, Bellefonte, PA, USA). The mobile phase was prepared according to Ruttanapattanakul et al. [22]. For the detection, gradient elution was done using two solvents, A [1% (v/v) acetic acid in water] and B (100% acetonitrile). Sample (10 µL) was injected with an initial flow rate of 1.0 mL/min and detection at 285 nm. Gradient elution mode was used conditions was according Jirakiattikul et al. [23] at a volume ratio of 80% A in 5 min, then reduced to 65% A in 5 min, remained at 0% A for 4 min, and then raised to 40% A in 5 min before returning to 80% A in 6 min. The overall run time per a sample was 25 min. The pinostobin and panduratin A calibration curve was developed at concentrations of 50 to 1000 mg/mL. The pinostobin and panduratin A content was reported in milligrams equivalents per dried sample.

2.5.6. Non-Volatile Metabolite Profiling Using Liquid Chromatography-Mass Spectrometer (LC-MS)

The identification of the soluble metabolites in the B. rotunda samples was investigated according with Sunanta, Chung, Kunasakdakul, Ruksiriwanich, Jantrawut, Hongsibsong, Sommano and Nutrition [21] using an Agilent 6545 QTOF-LC/MS system consisting of an Agilent 1260 LC with a 6545 UHD accurate-mass QTOF mass spectrometer. The crude extract of 0.01 mg was redissolved with 1.0 mL of LCMS-grade methanol (v/v) and filtered through a 0.25 m nylon syringe membrane filter. Separation of the chemical compounds was performed using the Agilent Poroshell 120 EC-C18 column (2.1 mm × 100 mm, 2.7 µm) at 25 °C. The mobile phase consisted of 0.1% (v/v) formic acid dissolved in water (phase A) and 0.1% (v/v) formic acid dissolved in methanol (phase B). The gradient elution was run as follows: 5 percent phase B for 1 min, increasing to 95 percent for 10 min, and then maintained for 19 min. The total run time was 30 min. The applied flow rate was 0.4 mL/min and the injection volume was 10 µL. MS and MS/MS analyses were done using a 6545 Agilent Ultra-High-Definition Accurate-Mass QTOF-MS linked to the LC, outfitted with an Agilent Dual Jet Stream electrospray ionization (Dual AJS ESI) interface in positive ionization mode, and Mass Hunter software was utilized for data elaboration (Agilent Technologies, Santa Clara, CA, USA). The correct mass and molecular formula prediction were screened against the database for hypothetical compounds (Agilent METLIN PCDL, Santa Clara, CA, USA).

2.5.7. Volatile Metabolite Profiling Using Pyrolysis Gas Chromatography Mass Spectrometry (Py-GC-MS)

Py-GC is a method of chemical analysis in which organic material is heated and thermochemically decomposed without participation of oxygen to produce smaller molecules that are separated by gas chromatography and detected using mass spectrometry [24,25]. The identification of volatile organic compounds in the B. rotunda powder was by the adapted method from Fakhrudin et al. [26] using an Agilent 7890B GC instrument. The powder samples 0.1 mg were placed in a pyrofoil (255 °C pyrolysis temperatures). The determination of volatile organic compounds was done using the GC (JCI-77pyrolysis probe, Japan Analytical Industry Co., Ltd., Tokyo, Japan). The column for gas chromatography was Rxi®-1 ms (30 m, 0.25 mm ID, and 0.25 µm df) produced by Restek (Cat. Number #13323; Bellefonte, PA, USA) with the FID detector set to 330 °C. The column conditions are as follows: Column oven temperature: 80 °C; injector temperature: 260 °C; and carrier gas Helium, constant flow rate 2.0 mL/min. The column temperature program: Initial temperature of 80 °C for 1 min and then increased 20 °C/min until the final temperature of 320 °C (maintained for 3 min). GC/MS data were processed with the ChemStation software (Agilent Technologies, Santa Clara, CA, USA) and the NIST 05 mass spectra library (Agilent, Santa Clara, CA, USA)

2.6. Statistical Analysis

The analyses of physical and chemical properties were carried out at least in biological and technical triplicates. Data was analyzed using one-way analysis of variance and Duncan’s test. Differences in values were considered significantly different when the p-value was <0.05 at the significant level of 95%. All statistical analysis was performed using IBM SPSS program v. 20. Physiological and physicochemical data among the samples were analyzed using principal component analysis (PCA). The RAPD data were displayed on dendrogram using the NTSYSpc2.2 program (Applied Biostatistics Inc., New York, NY, USA). The expression-based heat maps of metabolite compounds obtained from LC-MS and GC-MS data were performed using Heatmapper.ca (Creative Commons Attribution-ShareAlike 2.0 Generic).

3. Results and Discussion

3.1. Morphology

For morphological characteristics of the aerial parts, leaf growth as determined by the expanding of width and length of the MMC sample was noticed after one week after cultivation, while such characteristics of the DM1C, DM2C, SM2C, and MLW samples occurred at 3 weeks after cultivation. The leaf structure of the SM1C and MPW innitiated at 5 weeks after cultivation. All samples exhibited constant leaf growth at 9 weeks after cultivation before entering the dormant phase after 17 weeks (Figure 1a,b) In terms of stem growth, the MMC sample had stem growth as detemonstrated by expanding of the width and height was detected one week after cultivation. The stem growth of DM1C, DM2C, SM1C, SM2C, and MLW samples occurred at 3 weeks after cultivation, while this growth in MPW occurred at 5 weeks. The height of MLW samples was subtle after 11 weeks after cultivation while all plants stopped to expand after 15 weeks (Figure 1c,d) and thereafter entering the dormant period at 17 weeks. The morphological data of the aboveground parts are shown in Table 1. Overall, it appeared that MMC, and SM2C had the highest survivor rate (100%). MMC illustrated the biggest structure, viz., 8.03 cm of leaf width, 25.43 cm of leaf length, and 51.75 cm of stem height. However, the stem width of all samples was not significantly different (~1.06 cm). The morphology of the B. rotunda rhizome is shown in Table 2. It was found that the rhizome part of the DM1C and SM2C had the highest weight (~14.79 g), where as MMC had the highest finger weight (47.07 g). Structurelly, the rhizome length and finger length of all samples were not significantly different (~15.5 cm of rhizome length and~ 6 cm of finger length).
The distribution of the morphological data of all samples was depicted using multivariate PCA. For physical characteristics of the aboveground organs (Figure 2a,b), the PCA space accounted for 87.06% in PC1 and 8.71% in PC2. Based on the observation, the data were divided into two main clusters with the first cluster containing the samples of the cultivated type (MMC, DM1C, and DM2C) illustrating the larger structures of the leaf and stem compared to other samples as colored in blue. The second cluster contained those belonging to the wild types with smaller forms as indicated in pink. It was interesting that some of the cultivated type fell within these color regions (viz., those from the MLW, SM1C, and SM2C). The habitat information advised that these cultivated samples were previously from the wild seeding that had begun to adapt to the environment. The relationship of the physical characteristics of the underground organs were demonstrated in Figure 2c,d. The PCA space distributes 38.39% in PC1 and 33.00% in PC2. Based on the observation, data were clustered into two groups, the first cluster contained samples of the cultivated type (MMC, DM1C and DM2C) with larger underground structures as compared to other samples. The second cluster contains samples of the wild type (MLW and MPW), while SM1C, SM2C are those of the adapting type. All in all, the results showed that the physical characteristics of the aboveground organs corresponded to the physical characteristics of the rhizome. This finding is in line with Casper and Jackson [27], who advised that, although it was commonly assumed, shoot and root competition influenced plant growth characteristics. In addition, plant habitat often exerts its greatest influence on the characteristic of the aerial parts by altering leaf/stem ratios along with other morphological modifications [28]. During environmental stress, the expression of a variety of genes is induced which greatly influences plant growth and productivity [29]. Li et al. [30] also added that the environmental variables acted in a combinatorial fashion, best-explaining outlier genetic variation, and indicating environmentally dependent local adaptation for both leading- and trailing-edge populations. In the Zingiberaceae family, it is also found that plants exhibits adaptive evolution under different environmental conditions according to their phylogenetic relations [31]. Soil moisture is one of the most important constraints in cultivation of plants within the ginger family [32,33]. The moisture affects the accumulation of dry materials and the transportation of nitrogen in plants [34]. Contradictory, Osakabe, Osakabe, Shinozaki, and Tran [34] also pointed out that water stress gave the adverse impacts on plant physiology, especially the photosynthetic capacity. Plants have evolved complex physiological and biochemical adaptations to adjust and adapt to a variety of environmental stresses, thereby inducing the biosynthesis of the secondary metabolites. Beside this, plant genetics also contribute to phenotypic variation [35].

3.2. Genetic Variation of B. rotunda

Usually, different habitats and environmental factors have always had an impact on the observable traits. In addition, it is possible to mistakenly identify two distinct sympatric species with the same personal characteristics as one species [36]. The Boesenbergia is one of the consistently difficult genera within this family taxonomically, due to the heterogeneity of the physical characteristic at both the intra- and interspecific levels. As a result, molecular markers are necessary to clearly identify the correct species [19,37]. The RAPD-PCR fingerprints were generated from DNA extract from the fingerroot tissue of B. rotunda samples with 10 oligonucleotide primers, as shown in Table 1. The results indicated that there were 74 fragments based on the RAPD (Figure 3a), which were used to create a dendrogram, as shown in Figure 3b. The dendrogram categorized the samples into three groups. DM1C was projected away from the others, while the first cluster consisted of samples of cultivated-type (MMC and DM2C) and wild-type MLW having more than 70% similarity. The second cluster contained samples of wild type (MPW, SM1C, and SM2C) which exhibited more than 68% similarity. From the results, we then are able to assume the genetic linkage between the samples collected in the wild and the cultivated types. Though, when considering their morphological characteristics, they were clearly distinctive, while the SM1C and SM2C were within the midrange. Ebrahimi et al. [38] explained that variables in a gene pool could be as a result of plant adaptation to their natural habitat. However, the genetic similarity of the cultivated B. rotunda (Krachai Ban) and the wild type (Krachai Pah) with the identical morphological characteristics of having a small rhizome, green color on both sides of lamina and midrib, had been found [39]. The genome assembly of B. rotunda (2n = 36) of the vegetatively propagated species illustrated an unusually high heterozygosity of 3.01%, advising that the cultivated species may have undergone whole-genome duplication events or hybrid origin [40].

3.3. Phytochemical Compositions of B. rotunda

The phytoconstituent analysis of B. rotunda rhizomes from different sources are shown in Table 3. The highest contents of pinostrobin and panduratin A were found in the SM2C (43.27 ± 5.15 and 7.49 ± 1.11 mg/g dried sample), followed by the cultivated types DM1C and MLW. The MMC, DM2C and MPW were those that illustrated the lower contents of these phytochemicals. The highest contents of total phenolic constituents were found in the DM2C (23.79 ± 1.15 mg/g dried sample), followed by MPW, MMC, MLW, and SM2C. Nonetheless, the highest contents of total flavonoid constituents were found in the MPW (95.12 ± 31.00 mg/g dried sample), followed by the cultivated types MMC, DM2C, SM1C, DM1C, and SM2C. The antioxidant potential as illustrated by DPPH was rather low in all samples (~25.00%), while high potency as determined by the ABTS (>92.00%) was shown. The relationships of these phytochemical compositions among the B. rotunda samples were evaluated on PCA panels. The PCA score plots depicted > 70% of the data within PC1 and PC2 (Figure 4). The biplot analysis showed a relationship between the contents of pinostrobin and panduratin A and the DM1C, SM2C, and MLW sources. MMC, DM2C, and MPW were projected closely to the antioxidant activities as well as the total phenolic and flavonoid levels. However, there was no correlation between the SM1C and the bioactive potency. The projection patterns from the PCA were divergent from the previous results of the morphological traits. We can then assume that the morphological characteristics may not be able to describe the amount of the bioactive ingredients contained in the rhizome.
B. rotunda is a potential source of bioactive compounds flavonoids including alpinetin, boesenbergin, cardamonin, geraniol, krachaizin, panduratin, pinostrobin, pinocembrin, rotundaflavone, and silybin that showed many interesting pharmacological activities, such as antifungal, antibacterial, antioxidant, etc. Interestingly [41,42], polyphenols including caffeic acid, coumaric acid, chlorogenic acid, hesperidin, kaempferol, naringen, and quercetin [8], flavones, and cyclohexenyl chalcone derivatives. Several of these secondary metabolites have shown promising antiviral and anticancer activities [43,44]. Cardamonin and pinostrobin chalcone were found to be the most potential natural compounds against breast cancer cell line MDA-MB-231 and colon cancer (HT-29, HL-60) cell line [45,46]. Pinostrobin (5-hydroxy-7-methoxy flavanone) is a naturally occurring dietary bioflavonoid [47,48,49]. Many pharmacological activities are attributed to pinostrobin, viz., anti-oxidative [50], anti-inflammatory activity [51], anti-cancer [52], anti-viral activity [53], anti-Alzheimer [54], gastroprotective activity, quinone reductase-inducer activity, antioxidant activity, anti-microbial activity, anti-nociceptive activity, and anti-fungal activity [55]. Panduratin A is a natural chalcone compound [56] that exhibits a potent inhibitory effect against SARS-CoV-2 infection at both the pre-entry and post-infection phases. Also, it suppresses viral infectivity in human airway epithelial cells [7] and inhibits the growth of MCF-7 human breast cancer and HT-29 human colon adenocarcinoma cells [57].

3.4. Metabolite Profiling of B. rotunda

The results from the LC-MS illustrated 50 different metabolites of the nonvolatile organic compound and are listed in Supplementary Table S3. The heat map categorized the samples into two clusters (Figure 5a). The first cluster consisted of samples of the cultivated-type (SM1C and MMC) and wild-type MLW. The second cluster contained samples of the cultivated-type (SM2C, DM1C, and DM2C) and wild-type MPW. The results of the volatile organic compound profiling with the GC-MS are illustrated in Supplementary Table S4. The heat map categorized the samples into three groups (Figure 5b). MLW was projected away from the others, while the first cluster consisted of samples of the cultivated-type (SM2C and MMC) and wild-type MPW. The second cluster contained samples of the cultivated type (SM1C, DM1C, and DM2C). From the results, we are able to extrapolate the genetic relationship between the samples collected in the wild and cultivated types. As said, environmental factors manipulate the different genetic expression the secondary metabolites biosynthesis pathways [58,59,60,61]. Plant secondary metabolites are not only a useful array of natural products but also an important part of plant defense systems against biotic and abiotic stresses [61]. Verma and Shukla [62] explained that the content of different secondary metabolites varied from generation to generation, and it did not remain stable because several biotic and abiotic factors play an important role in their synthesis and regulation.

4. Conclusions

This work evaluated the relationship between phenotypes and chemical profiling of B. rotunda collected from different habitats of Northern Thailand. Wild and domesticated, the findings of this study suggested that the morphological characteristics of the plants were not directly correlated with the levels of significant phytochemicals present, including pinostrobin and panduratin A, but rather were influenced by various agronomic factors. The random amplification of polymorphic DNA with ten base-pair primers indicates the genetic correlation between wild and cultivated types. The results of this study could assist plant breeders in selecting lines with favorable phenotypes, such as those with the largest biomass, under optimal agronomic conditions. The potential trait of fingerroot might thereafter be employed as a source of medicinal plants for commercial production of bioactive products for the prevention of SARS-CoV-2 infection. Further research is necessary to identify the most suitable conditions for the biosynthesis of the active compounds.

Supplementary Materials

The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/agronomy13061435/s1, Supplementary Table S1. Random amplified polymorphic DNA primer sequence of genetic variation experiment. Supplementary Table S2. Reaction compounds (reaction mix and template DNA). Supplementary Table S3. Metabolite compounds by using LC-MS of B. rotunda rhizome extracts. Supplementary Table S4. Metabolite compounds by using GC-MS of B. rotunda rhizome extracts.

Author Contributions

Conceptualization, S.R.S.; methodology, S.R.S., C.L. and M.W.; validation, S.R.S. and M.W.; formal analysis, S.T.; investigation, S.T. and S.R.S.; writing—original draft preparation, S.T. and M.W.; writing—review and editing, S.R.S., S.T. and M.W.; supervision, S.R.S., P.J., M.W. and W.B.; project administration, S.T.; funding acquisition, S.R.S. and M.W. All authors have read and agreed to the published version of the manuscript.

Funding

This research project was supported by the IRTC and was partially supported by Chiang Mai University.

Data Availability Statement

Not applicable.

Acknowledgments

We would like to especially thank Chiang Mai University, Thailand, for the support and infrastructure provided for the conduct of this research.

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. Tewtrakul, S.; Subhadhirasakul, S. Anti-allergic activity of some selected plants in the Zingiberaceae family. J. Ethnopharmacol. 2007, 109, 535–538. [Google Scholar] [CrossRef] [PubMed]
  2. Cheenpracha, S.; Karalai, C.; Ponglimanont, C.; Subhadhirasakul, S.; Tewtrakul, S. Anti-HIV-1 protease activity of compounds from Boesenbergia pandurata. Bioorg. Med. Chem. 2006, 14, 1710–1714. [Google Scholar] [CrossRef] [PubMed]
  3. Yuliana, N.D.; Budijanto, S.; Verpoorte, R.; Choi, Y.H.J. NMR metabolomics for identification of adenosine A1 receptor binding compounds from Boesenbergia rotunda rhizomes extract. J. Ethnopharmacol. 2013, 150, 95–99. [Google Scholar] [CrossRef]
  4. Chomchalow, N.; Bansiddhi, J.; MacBaine, C. Amazing Thai Medicinal Plants; Horticultural Research Institute Department of Agricultural, and Horticultural Science Society of Thailand: Bangkok, Thailand, 2003. [Google Scholar]
  5. Chuakul, W.; Boonpleng, A. Ethnomedical Uses of Thai Zingiberaceous Plant (1); Mahidol University: Bangkok, Thailand, 2003. [Google Scholar]
  6. Mood, J.; Tanaka, N.; Aung, M.; Murata, J. The genus Boesenbergia (Zingiberaceae) in Myanmar with two new records. Gard. Bull. Singap. 2016, 68, 299–318. [Google Scholar] [CrossRef]
  7. Kanjanasirirat, P.; Suksatu, A.; Manopwisedjaroen, S.; Munyoo, B.; Tuchinda, P.; Jearawuttanakul, K.; Seemakhan, S.; Charoensutthivarakul, S.; Wongtrakoongate, P.; Rangkasenee, N. High-content screening of Thai medicinal plants reveals Boesenbergia rotunda extract and its component Panduratin A as anti-SARS-CoV-2 agents. Sci. Rep. 2020, 10, 19963. [Google Scholar] [CrossRef]
  8. Jing, L.J.; Mohamed, M.; Rahmat, A.; Bakar, M.F.A. Phytochemicals, antioxidant properties and anticancer investigations of the different parts of several gingers species (Boesenbergia rotunda, Boesenbergia pulchella var attenuata and Boesenbergia armeniaca). J. Med. Plants 2010, 4, 27–32. [Google Scholar]
  9. Ng, T.L.M.; Karim, R.; Tan, Y.S.; Teh, H.F.; Danial, A.D.; Ho, L.S.; Khalid, N.; Appleton, D.R.; Harikrishna, J.A. Amino acid and secondary metabolite production in embryogenic and non-embryogenic callus of fingerroot ginger (Boesenbergia rotunda). PLoS ONE 2016, 11, e0156714. [Google Scholar] [CrossRef]
  10. Eng-Chong, T.; Yean-Kee, L.; Chin-Fei, C.; Choon-Han, H.; Sher-Ming, W.; Li-Ping, C.T.; Gen-Teck, F.; Khalid, N.; Abd Rahman, N.; Karsani, S.A.J.E.-B.C.; et al. Boesenbergia rotunda: From ethnomedicine to drug discovery. Evid.-Based Complement. Altern. Med. 2012, 2012, 473637. [Google Scholar] [CrossRef]
  11. Bhamarapravati, S.; Juthapruth, S.; Mahachai, W.; Mahady, G.J.S. Antibacterial activity of Boesenbergia rotunda (L.) Mansf. and Myristica fragrans Houtt. against Helicobacter pylori. Songklanakarin J. Sci. Technol. 2006, 28, 157–163. [Google Scholar]
  12. Hwang, J.-K.; Chung, J.-Y.; Baek, N.-I.; Park, J.-H. Isopanduratin A from Kaempferia pandurata as an active antibacterial agent against cariogenic Streptococcus mutans. Int. J. Antimicrob. Agents 2004, 23, 377–381. [Google Scholar] [CrossRef]
  13. Jaipetch, T.; Kanghae, S.; Pancharoen, O.; Patrick, V.; Reutrakul, V.; Tuntiwachwuttikul, P.; White, A. Constituents of Boesenbergia pandurata (syn. Kaempferia pandurata): Isolation, crystal structure and synthesis of (±)-Boesenbergin A. Aust. J. Chem. 1982, 35, 351–361. [Google Scholar] [CrossRef]
  14. Noor Diyana, M.M. Flavonoid Pathway Gene Discoveries in Boesenbergia Rotunda through RNA-SEQ Transcriptome Profiling of Cell Suspension Cultures in Response to Phenylalanine/Noor Diyana Md Mustafa. Doctoral Dissertation, University of Malaya, Kuala Lumpur, Malaysia, 2017. [Google Scholar]
  15. Kirana, C.; Record, I.R.; McIntosh, G.H.; Jones, G.P. Screening for antitumor activity of 11 species of Indonesian zingiberaceae using human MCF-7 and HT-29 cancer cells. Pharm. Biol. 2003, 41, 271–276. [Google Scholar] [CrossRef]
  16. Kiat, T.S.; Pippen, R.; Yusof, R.; Ibrahim, H.; Khalid, N.; Abd Rahman, N. Inhibitory activity of cyclohexenyl chalcone derivatives and flavonoids of fingerroot, Boesenbergia rotunda (L.), towards dengue-2 virus NS3 protease. Bioorga. Med. Chem. Lett. 2006, 16, 3337–3340. [Google Scholar] [CrossRef]
  17. Watanaponpan, S.; Sairuksa, D.; Singhalert, R. White Galingale Cultivation for Promoting Community Economy of Farmers in Tha Maka District, Kanchanaburi Province. Interdiscip. Acad. Res. J. 2022, 2, 499–512. [Google Scholar]
  18. Prathepha, P. Screening of random primer to evaluate DNA diversity in Thai Curcuma using random amplified polymorphic DNAs. Warasan Songkhlanakarin 2000, 22, 7–13. [Google Scholar]
  19. Vanijajiva, O.; Sirirugsa, P.; Suvachittanont, W. Confirmation of relationships among Boesenbergia (Zingiberaceae) and related genera by RAPD. Biochem. Syst. Ecol. 2005, 33, 159–170. [Google Scholar] [CrossRef]
  20. Sangta, J.; Wongkaew, M.; Tangpao, T.; Withee, P.; Haituk, S.; Arjin, C.; Sringarm, K.; Hongsibsong, S.; Sutan, K.; Pusadee, T.J.P. Recovery of polyphenolic fraction from arabica coffee pulp and its antifungal applications. Plants 2021, 10, 1422. [Google Scholar] [CrossRef]
  21. Sunanta, P.; Chung, H.H.; Kunasakdakul, K.; Ruksiriwanich, W.; Jantrawut, P.; Hongsibsong, S.; Sommano, S.R. Genomic relationship and physiochemical properties among raw materials used for Thai black garlic processing. Food Sci. Nutr. 2020, 8, 4534–4545. [Google Scholar] [CrossRef]
  22. Ruttanapattanakul, J.; Wikan, N.; Okonogi, S.; Takuathung, M.N.; Buacheen, P.; Pitchakarn, P.; Potikanond, S.; Nimlamool, W. Boesenbergia rotunda extract accelerates human keratinocyte proliferation through activating ERK1/2 and PI3K/Akt kinases. Biomed. Pharmacother. 2021, 133, 111002. [Google Scholar] [CrossRef]
  23. Jirakiattikul, Y.; Rithichai, P.; Prachai, R.; Itharat, A. Elicitation enhancement of bioactive compound accumulation and antioxidant activity in shoot cultures of Boesenbergia rotunda L. Agric. Nat. Resour. 2021, 55, 456–463. [Google Scholar]
  24. Picó, Y.; Barceló, D. Pyrolysis gas chromatography-mass spectrometry in environmental analysis: Focus on organic matter and microplastics. TrAC Trends Anal. Chem. 2020, 130, 115964. [Google Scholar] [CrossRef]
  25. Chiavari, G.; Galletti, G.C.; Lanterna, G.; Mazzeo, R. The potential of pyrolysis—Gas chromatography/mass spectrometry in the recognition of ancient painting media. J. Anal. Appl. Pyrolysis 1993, 24, 227–242. [Google Scholar] [CrossRef]
  26. Fakhrudin, N.; Wiyono, T.; Putra, A.R.; Nurrochmad, A.; Widyarini, S.J.T. The evaluation on anti-platelet and antithrombosis activities of Cinnamomum sintoc bark extract. Thai J. Pharm. Sci. 2020, 43, 219–226. [Google Scholar]
  27. Casper, B.B.; Jackson, R.B. Plant competition underground. Annu. Rev. Ecol. Syst. 1997, 28, 545–570. [Google Scholar] [CrossRef]
  28. Buxton, D.R.; Fales, S.L. Plant environment and quality. Forage Qual. Eval. Util. 1994, 155–199. [Google Scholar] [CrossRef]
  29. Yamaguchi-Shinozaki, K.; Shinozaki, K. Transcriptional regulatory networks in cellular responses and tolerance to dehydration and cold stresses. Annu. Rev. Plant Biol. 2006, 57, 781–803. [Google Scholar] [CrossRef]
  30. Li, Y.-S.; Liao, P.-C.; Chang, C.-T.; Hwang, S.-Y. Pattern of Adaptive Divergence in Zingiber kawagoii Hayata (Zingiberaceae) along a Narrow Latitudinal Range. Plants 2022, 11, 2490. [Google Scholar] [CrossRef]
  31. Gao, B.; Yuan, L.; Tang, T.; Hou, J.; Pan, K.; Wei, N. The complete chloroplast genome sequence of Alpinia oxyphylla Miq. and comparison analysis within the Zingiberaceae family. PLoS ONE 2019, 14, e0218817. [Google Scholar] [CrossRef]
  32. Bresson, J.; Vasseur, F.; Dauzat, M.; Koch, G.; Granier, C.; Vile, D. Quantifying spatial heterogeneity of chlorophyll fluorescence during plant growth and in response to water stress. Plant Methods 2015, 11, 23. [Google Scholar] [CrossRef]
  33. Li, H.; Huang, M.; Tan, D.; Liao, Q.; Zou, Y.; Jiang, Y. Effects of soil moisture content on the growth and physiological status of ginger (Zingiber officinale Roscoe). Acta Physiol. Plant. 2018, 40, 125. [Google Scholar] [CrossRef]
  34. Osakabe, Y.; Osakabe, K.; Shinozaki, K.; Tran, L.-S.P. Response of plants to water stress. Front. Plant Sci. 2014, 5, 86. [Google Scholar] [CrossRef]
  35. Hallauer, A.R. Evolution of plant breeding. Crop Breed. Appl. Biotechnol. 2011, 11, 197–206. [Google Scholar] [CrossRef]
  36. Techaprasan, J.; Ngamriabsakul, C.; Klinbunga, S.; Chusacultanachai, S.; Jenjittikul, T. Genetic variation and species identification of Thai Boesenbergia (Zingiberaceae) analyzed by chloroplast DNA polymorphism. BMB Rep. 2006, 39, 361–370. [Google Scholar] [CrossRef]
  37. Techaprasan, J.; Klinbunga, S.; Jenjittikul, T. Genetic relationships and species authentication of Boesenbergia (Zingiberaceae) in Thailand based on AFLP and SSCP analyses. Biochem. Syst. Ecol. 2008, 36, 408–416. [Google Scholar] [CrossRef]
  38. Ebrahimi, F.; Mohammadi-Nejad, G.; Baghizadeh, A.; Abdolinejad, M. Genetic diversity evaluation of rapeseed genotypes (Brassica napus L.) based on phenotypic traits and random amplified polymorphic DNA (RAPD) markers. Afr. J. Biotechnol. 2011, 10, 17391–17398. [Google Scholar] [CrossRef]
  39. Theanphong, O.; Mingvanish, W.; Jenjittikul, T.; Rungsihirunrat, K. Genetic variation of Boesenbergia rotunda (L.) Mansf. from Thailand based on essential oil compositions and internal transcribed spacer sequences. Songklanakarin J. Sci. Technol. 2021, 43, 1620–1627. [Google Scholar]
  40. Taheri, S.; Teo, C.H.; Heslop-Harrison, J.S.; Schwarzacher, T.; Tan, Y.S.; Wee, W.Y.; Khalid, N.; Biswas, M.K.; Mutha, N.V.R.; Mohd-Yusuf, Y.; et al. Genome Assembly and Analysis of the Flavonoid and Phenylpropanoid Biosynthetic Pathways in Fingerroot Ginger (Boesenbergia rotunda). Int. J. Mol. Sci. 2022, 23, 7269. [Google Scholar] [CrossRef]
  41. Gurung, A.B.; Ali, M.A.; Al-Hemaid, F.; El-Zaidy, M.; Lee, J. In silico analyses of major active constituents of fingerroot (Boesenbergia rotunda) unveils inhibitory activities against SARS-CoV-2 main protease enzyme. Saudi J. Biol. Sci. 2022, 29, 65–74. [Google Scholar] [CrossRef]
  42. Chahyadi, A.; Hartati, R.; Wirasutisna, K.R.J.P.C. Boesenbergia pandurata Roxb., an Indonesian medicinal plant: Phytochemistry, biological activity, plant biotechnology. Procedia Chem. 2014, 13, 13–37. [Google Scholar] [CrossRef]
  43. Saah, S.; Siriwan, D.; Trisonthi, P. Biological activities of Boesenbergia rotunda parts and extracting solvents in promoting osteogenic differentiation of pre-osteoblasts. Food Biosci. 2021, 41, 101011. [Google Scholar] [CrossRef]
  44. Wong, S.M.; Salim, N.; Harikrishna, J.A.; Khalid, N. Highly efficient plant regeneration via somatic embryogenesis from cell suspension cultures of Boesenbergia rotunda. Cell. Dev. Biol.-Plant 2013, 49, 665–673. [Google Scholar] [CrossRef]
  45. Mohammed, I.A.; Akhtar, M.N.; Biau, F.J.; Tor, Y.S.; Zareen, S.; Binti Shahabudin, S.; Binti Abd Hamid, H.; Ul Haq, Z.; Khalil, R.; Khalaf, R.M. Isolation of cardamonin and pinostrobin chalcone from the rhizomes of Boesenbergia rotunda (L.) Mansf. and their cytotoxic effects on H-29 and MDA-MB-231 cancer cell lines. Nat. Prod. J. 2019, 9, 341–348. [Google Scholar] [CrossRef]
  46. Yap, A.L.C. Phytochemical and Hplc Profiling of Extracts from Fingerroot (Boesenbergia rotunda) Rhizomes. Doctoral Dissertation, Universiti Putra Malaysia, Selangor, Malaysia, 2008. [Google Scholar]
  47. Jaudan, A.; Sharma, S.; Malek, S.N.A.; Dixit, A.J.P.O. Induction of apoptosis by pinostrobin in human cervical cancer cells: Possible mechanism of action. PLoS ONE 2018, 13, e0191523. [Google Scholar] [CrossRef] [PubMed]
  48. Gu, C.; Fu, L.; Yuan, X.; Liu, Z.J.M. Promoting effect of pinostrobin on the proliferation, differentiation, and mineralization of murine pre-osteoblastic MC3T3-E1 cells. Molecules 2017, 22, 1735. [Google Scholar] [CrossRef] [PubMed]
  49. Athapaththu, A.M.G.K.; Lee, K.T.; Kavinda, M.H.D.; Lee, S.; Kang, S.; Lee, M.-H.; Kang, C.-H.; Choi, Y.H.; Kim, G.-Y. Pinostrobin ameliorates lipopolysaccharide (LPS)-induced inflammation and endotoxemia by inhibiting LPS binding to the TLR4/MD2 complex. Biomed. Pharmacother. 2022, 156, 113874. [Google Scholar] [CrossRef]
  50. Christov, R.; Trusheva, B.; Popova, M.; Bankova, V.; Bertrand, M. Chemical composition of propolis from Canada, its antiradical activity and plant origin. Nat. Prod. Res. 2006, 20, 531–536. [Google Scholar] [CrossRef]
  51. Patel, N.K.; Bhutani, K.K.J.P. Pinostrobin and Cajanus lactone isolated from Cajanus cajan (L.) leaves inhibits TNF-α and IL-1β production: In vitro and in vivo experimentation. Phytomedicine 2014, 21, 946–953. [Google Scholar] [CrossRef]
  52. Le Bail, J.-C.; Aubourg, L.; Habrioux, G. Effects of pinostrobin on estrogen metabolism and estrogen receptor transactivation. Cancer Lett. 2000, 156, 37–44. [Google Scholar] [CrossRef]
  53. Wu, N.; Kong, Y.; Zu, Y.; Fu, Y.; Liu, Z.; Meng, R.; Liu, X.; Efferth, T.J.P. Activity investigation of pinostrobin towards herpes simplex virus-1 as determined by atomic force microscopy. Phytomedicine 2011, 18, 110–118. [Google Scholar] [CrossRef]
  54. Xian, Y.-F.; Ip, S.-P.; Lin, Z.-X.; Mao, Q.-Q.; Su, Z.-R.; Lai, X.-P. Protective effects of pinostrobin on β-amyloid-induced neurotoxicity in PC12 cells. Cell. Mol. Neurobiol. 2012, 32, 1223–1230. [Google Scholar] [CrossRef]
  55. Patel, N.K.; Jaiswal, G.; Bhutani, K.K. A review on biological sources, chemistry and pharmacological activities of pinostrobin. Nat. Prod. Res. 2016, 30, 2017–2027. [Google Scholar] [CrossRef]
  56. Rukayadi, Y.; Han, S.; Yong, D.; Hwang, J.-K. In vitro antibacterial activity of panduratin A against enterococci clinical isolates. Biol. Pharm. Bull. 2010, 33, 1489–1493. [Google Scholar] [CrossRef]
  57. Kirana, C.; Jones, G.P.; Record, I.R.; McIntosh, G.H.J. Anticancer properties of panduratin A isolated from Boesenbergia pandurata (Zingiberaceae). J. Nat. Med. 2007, 61, 131–137. [Google Scholar] [CrossRef]
  58. Ashraf, M.A.; Iqbal, M.; Rasheed, R.; Hussain, I.; Riaz, M.; Arif, M.S. Environmental stress and secondary metabolites in plants: An overview. Plant Metab. Regul. Environ. Stress 2018, 153–167. [Google Scholar]
  59. Borges, C.V.; Minatel, I.O.; Gomez-Gomez, H.A.; Lima, G.P.P. Medicinal plants: Influence of environmental factors on the content of secondary metabolites. In Medicinal Plants and Environmental Challenges; Springer: Berlin/Heidelberg, Germany, 2017; pp. 259–277. [Google Scholar]
  60. Dey, M.; Kalia, S.; Ghosh, S.; Guha-Mukherjee, S. Biochemical and molecular basis of differentiation in plant tissue culture. Curr. Sci. 1998, 74, 591–596. [Google Scholar]
  61. Yang, L.; Wen, K.-S.; Ruan, X.; Zhao, Y.-X.; Wei, F.; Wang, Q.J.M. Response of plant secondary metabolites to environmental factors. Molecules 2018, 23, 762. [Google Scholar] [CrossRef]
  62. Verma, N.; Shukla, S. Impact of various factors responsible for fluctuation in plant secondary metabolites. J. Appl. Res. Med. Aromat. Plants 2015, 2, 105–113. [Google Scholar] [CrossRef]
Agronomy 13 01435 g001 550
Figure 1. Morphological characteristics of B. rotunda after cultivated for 17 weeks. (a) leaf width (b) leaf length (c) stem height and (d) stem width. Data are means of the replicate samples. Bars represent standard deviation and different alphabet letters denote significant differences (95%) among the samples of each week (p > 0.05). MMC = Mae Taeng, Chiang Mai, cultivated type; MLW = Mae Tha, Lamphun, wild type; DM1C = Chiang Dao1, Chiang Mai, cultivated type; DM2C = Chiang Dao2, Chiang Mai, cultivated type; SM1C = San Pa Tong1, Chiang Mai, cultivated type; SM2C = San Pa Tong2, Chiang Mai, cultivated type; MPW = Mae Moh, Lampang, wild type. Data expressed as mean ± standard deviation.
Figure 1. Morphological characteristics of B. rotunda after cultivated for 17 weeks. (a) leaf width (b) leaf length (c) stem height and (d) stem width. Data are means of the replicate samples. Bars represent standard deviation and different alphabet letters denote significant differences (95%) among the samples of each week (p > 0.05). MMC = Mae Taeng, Chiang Mai, cultivated type; MLW = Mae Tha, Lamphun, wild type; DM1C = Chiang Dao1, Chiang Mai, cultivated type; DM2C = Chiang Dao2, Chiang Mai, cultivated type; SM1C = San Pa Tong1, Chiang Mai, cultivated type; SM2C = San Pa Tong2, Chiang Mai, cultivated type; MPW = Mae Moh, Lampang, wild type. Data expressed as mean ± standard deviation.
Agronomy 13 01435 g001
Agronomy 13 01435 g002 550
Figure 2. The principal component (PC) analysis of the B. rotunda sample based on their physical characteristics. (a) Variables of aboveground organ; (b) observations of aboveground organ; (c) variables of underground organ; and (d) observations of underground organ. MMC = Mae Taeng, Chiang Mai, cultivated type; MLW = Mae Tha, Lamphun, wild type; DM1C = Chiang Dao1, Chiang Mai, cultivated type; DM2C = Chiang Dao2, Chiang Mai, cultivated type; SM1C = San Pa Tong1, Chiang Mai, cultivated type; SM2C = San Pa Tong2, Chiang Mai, cultivated type; MPW = Mae Moh, Lampang, wild type.
Figure 2. The principal component (PC) analysis of the B. rotunda sample based on their physical characteristics. (a) Variables of aboveground organ; (b) observations of aboveground organ; (c) variables of underground organ; and (d) observations of underground organ. MMC = Mae Taeng, Chiang Mai, cultivated type; MLW = Mae Tha, Lamphun, wild type; DM1C = Chiang Dao1, Chiang Mai, cultivated type; DM2C = Chiang Dao2, Chiang Mai, cultivated type; SM1C = San Pa Tong1, Chiang Mai, cultivated type; SM2C = San Pa Tong2, Chiang Mai, cultivated type; MPW = Mae Moh, Lampang, wild type.
Agronomy 13 01435 g002
Agronomy 13 01435 g003 550
Figure 3. Genetic variation analysis of B. rotunda samples obtained from different habitats using the RAPD-PCR technique. (a) The random amplified polymorphic DNA (RAPD)-PCR fingerprints using 10 random primers: OPAM-01 = TCACGTACGG, OPAM-03 = CTTCCCTGTG, OPAM-12 = TCTCACCGTC, OPAM-18 = ACGGGACTCT, OPA-18 = AGGTGACCGT, OPB-01 = GTTTCGCTCC, OPB-14 = TCCGCTCTGG, OPC-05 = GATGACCGCC, OPD-03 = GTCGCCGTCA, and OPD-08 = GTGTGCCCCA; (b) The dendrogram generated from the similarity matrix of RAPD data. MMC = Mae Taeng, Chiang Mai, cultivated type; MLW = Mae Tha, Lamphun, wild type; DM1C = Chiang Dao1, Chiang Mai, cultivated type; DM2C = Chiang Dao2, Chiang Mai, cultivated type; SM1C = San Pa Tong1, Chiang Mai, cultivated type; SM2C = San Pa Tong2, Chiang Mai, cultivated type; MPW = Mae Moh, Lampang, wild type.
Figure 3. Genetic variation analysis of B. rotunda samples obtained from different habitats using the RAPD-PCR technique. (a) The random amplified polymorphic DNA (RAPD)-PCR fingerprints using 10 random primers: OPAM-01 = TCACGTACGG, OPAM-03 = CTTCCCTGTG, OPAM-12 = TCTCACCGTC, OPAM-18 = ACGGGACTCT, OPA-18 = AGGTGACCGT, OPB-01 = GTTTCGCTCC, OPB-14 = TCCGCTCTGG, OPC-05 = GATGACCGCC, OPD-03 = GTCGCCGTCA, and OPD-08 = GTGTGCCCCA; (b) The dendrogram generated from the similarity matrix of RAPD data. MMC = Mae Taeng, Chiang Mai, cultivated type; MLW = Mae Tha, Lamphun, wild type; DM1C = Chiang Dao1, Chiang Mai, cultivated type; DM2C = Chiang Dao2, Chiang Mai, cultivated type; SM1C = San Pa Tong1, Chiang Mai, cultivated type; SM2C = San Pa Tong2, Chiang Mai, cultivated type; MPW = Mae Moh, Lampang, wild type.
Agronomy 13 01435 g003
Agronomy 13 01435 g004 550
Figure 4. The principal component (PC) analysis of the B. rotunda sample based on their phytochemical compositions. MMC = Mae Taeng, Chiang Mai, cultivated type; MLW = Mae Tha, Lamphun, wild type; DM1C = Chiang Dao1, Chiang Mai, cultivated type; DM2C = Chiang Dao2, Chiang Mai, cultivated type; SM1C = San Pa Tong1, Chiang Mai, cultivated type; SM2C = San Pa Tong2, Chiang Mai, cultivated type; MPW = Mae Moh, Lampang, wild type.
Figure 4. The principal component (PC) analysis of the B. rotunda sample based on their phytochemical compositions. MMC = Mae Taeng, Chiang Mai, cultivated type; MLW = Mae Tha, Lamphun, wild type; DM1C = Chiang Dao1, Chiang Mai, cultivated type; DM2C = Chiang Dao2, Chiang Mai, cultivated type; SM1C = San Pa Tong1, Chiang Mai, cultivated type; SM2C = San Pa Tong2, Chiang Mai, cultivated type; MPW = Mae Moh, Lampang, wild type.
Agronomy 13 01435 g004
Agronomy 13 01435 g005 550
Figure 5. Heat map plots clusters of B. rotunda samples based on their phytochemical compositions using (a) LC-MS and (b) GC-MS techniques based on the content. MMC = Mae Taeng, Chiang Mai, cultivated type; MLW = Mae Tha, Lamphun, wild type; DM1C = Chiang Dao1, Chiang Mai, cultivated type; DM2C = Chiang Dao2, Chiang Mai, cultivated type; SM1C = San Pa Tong1, Chiang Mai, cultivated type; SM2C = San Pa Tong2, Chiang Mai, cultivated type; MPW = Mae Moh, Lampang, wild type.
Figure 5. Heat map plots clusters of B. rotunda samples based on their phytochemical compositions using (a) LC-MS and (b) GC-MS techniques based on the content. MMC = Mae Taeng, Chiang Mai, cultivated type; MLW = Mae Tha, Lamphun, wild type; DM1C = Chiang Dao1, Chiang Mai, cultivated type; DM2C = Chiang Dao2, Chiang Mai, cultivated type; SM1C = San Pa Tong1, Chiang Mai, cultivated type; SM2C = San Pa Tong2, Chiang Mai, cultivated type; MPW = Mae Moh, Lampang, wild type.
Agronomy 13 01435 g005
Table
Table 1. Physical characteristics of B. rotunda aboveground organs at 17 weeks.
Table 1. Physical characteristics of B. rotunda aboveground organs at 17 weeks.
SampleCultivated
Area/GPS
Coordinates		HabitatSurvivor
Rate (%)		LeafStemsAbove
Ground Part
Width (cm)Length (cm)Width (cm)Height (cm)
MMCMae Taeng, Chiang Mai/19.05992, 98.83022cultivation type1008.03 ± 1.68 a25.43 ± 5.00 a1.45 ± 0.40 a51.75 ± 10.17 aAgronomy 13 01435 i001
MLWMae Tha, Lamphun/18.63397, 99.04039wild type505.57 ± 1.63 b15.89 ± 5.25 bc1.08 ± 0.41 cd30.46 ± 10.45 cAgronomy 13 01435 i002
DM1CChiang Dao, Chiang Mai/19.30270, 98.96640cultivation type957.28 ± 1.95 a17.71 ± 4.81 b1.29 ± 0.32 ab33.97 ± 11.35 bcAgronomy 13 01435 i003
DM2CChiang Dao, Chiang Mai/19.51182, 99.09041cultivation type807.25 ± 1.65 a23.23 ± 4.57 b1.33 ± 0.32 ab44.43 ± 10.02 cAgronomy 13 01435 i004
SM1CSan Pa Tong1, Chiang Mai/18.62599, 98.88333cultivation type505.74 ± 1.82 b17.28 ± 5.53 b0.96 ± 0.27 bc33.78 ± 12.78 cAgronomy 13 01435 i005
SM2CSan Pa Tong2, Chiang Mai/18.62599, 98.88367cultivation type1007.82 ± 2.04 a19.17 ± 5.52 a1.44 ± 0.45 a36.59 ± 12.72 abAgronomy 13 01435 i006
MPWMae Moh, Lampang/18.42797, 99.82055wild type454.30 ± 1.02 b12.42 ± 3.01 c0.67 ± 0.28 d21.40 ± 3.60 dAgronomy 13 01435 i007
MMC = Mae Taeng, Chiang Mai, cultivated type; MLW = Mae Tha, Lamphun, wild type; DM1C = Chiang Dao1, Chiang Mai, cultivated type; DM2C = Chiang Dao2, Chiang Mai, cultivated type; SM1C = San Pa Tong1, Chiang Mai, cultivated type; SM2C = San Pa Tong2, Chiang Mai, cultivated type; MPW = Mae Moh, Lampang, wild type. Data expressed as mean ± standard deviation. a–d Values in the same column with different alphabets are statistically distinct (p > 0.05).
Table
Table 2. Physical properties of B. rotunda underground organs at 18 weeks.
Table 2. Physical properties of B. rotunda underground organs at 18 weeks.
SampleMother RhizomesFingersRhizome
Weight (g)Width (cm)Length (cm)Weight (g)Width (cm)Length (cm)
MMC8.24 ± 2.66 ab14.49 ± 0.56 a17.61 ± 2.70 ns47.07 ± 31.69 a12.69 ± 0.79 a5.86 ± 0.97 nsAgronomy 13 01435 i008
MLW4.36 ± 2.84 b12.53 ± 0.28 b13.90 ± 1.03 ns20.79 ± 1.75 ab11.44 ± 3.24 ab6.50 ± 1.59 nsAgronomy 13 01435 i009
DM1C14.16 ± 5.61 a14.07 ± 0.89 a17.87 ± 2.60 ns37.53 ± 25.23 ab13.47 ± 0.39 a6.49 ± 2.32 nsAgronomy 13 01435 i010
DM2C3.23 ± 1.38 b12.13 ± 0.62 bc16.98 ± 3.43 ns23.51 ± 2.45 ab12.89 ± 2.49 a6.92 ± 1.04 nsAgronomy 13 01435 i011
SM1C2.84 ± 2.25 b10.06 ± 0.62 d13.71 ± 2.94 ns19.30 ± 3.11 ab12.73 ± 0.89 a5.94 ± 1.08 nsAgronomy 13 01435 i012
SM2C15.42 ± 7.63 a13.26 ± 0.39 ab17.84 ± 1.33 ns34.83 ± 24.39 ab13.96 ± 0.60 a6.83 ± 0.68 nsAgronomy 13 01435 i013
MPW2.99 ± 1.05 b11.18 ± 1.14 cd16.97 ± 4.45 ns4.87 ± 1.87 b9.31 ± 2.06 b5.32 ± 5.10 nsAgronomy 13 01435 i014
MMC = Mae Taeng, Chiang Mai, cultivated type; MLW = Mae Tha, Lamphun, wild type; DM1C = Chiang Dao1, Chiang Mai, cultivated type; DM2C = Chiang Dao2, Chiang Mai, cultivated type; SM1C = San Pa Tong1, Chiang Mai, cultivated type; SM2C = San Pa Tong2, Chiang Mai, cultivated type; MPW = Mae Moh, Lampang, wild type. Data expressed as mean ± standard deviation. a–d Values in the same column with different alphabets are statistically distinct (p > 0.05). ns not significant.
Table
Table 3. Phytoconstituents of B. rotunda from different sources.
Table 3. Phytoconstituents of B. rotunda from different sources.
SamplePinostrobin
(mg/gdried sample)		Panduratin A
(mg/gdried sample)		Total
Phenolic
(mggallic/gdried sample)		Total
Flavonoids
(mgcatechin/gdried sample)		DPPH
(%)		ABTS
(%)
MMC27.09 ± 6.97 bc4.90 ± 1.16 bc20.80 ± 2.64 ab73.71 ± 17.04 b30.76 ± 5.88 a92.25 ± 0.52 b
MLW30.05 ± 7.55 bc5.83 ± 0.58 ab19.69 ± 0.34 bc50.00 ± 10.82 c24.99 ± 2.33 ab92.59 ± 1.27 ab
DM1C35.38 ± 10.43 ab6.06 ± 1.50 ab16.97 ± 2.12 c57.46 ± 13.36 bc24.57 ± 2.73 ab93.28 ± 0.43 ab
DM2C27.56 ± 8.30 bc5.04 ± 1.49 bc23.79 ± 1.15 a65.62 ± 2.75 bc27.71 ± 3.47 a93.82 ± 0.39 a
SM1C14.41 ± 2.43 c3.08 ± 0.81 c16.52 ± 1.99 c63.23 ± 3.78 bc14.07 ± 5.45 b93.08 ± 1.44 ab
SM2C43.27 ± 5.15 a7.49 ± 1.11 a19.22 ± 1.92 bc54.99 ± 7.45 bc26.44 ± 4.60 a92.69 ± 1.39 ab
MPW17.27 ± 6.24 c3.10 ± 0.69 c22.88 ± 2.07 ab95.12 ± 31.00 a23.65 ± 2.68 ab93.96 ± 0.53 ab
MMC = Mae Taeng, Chiang Mai; MLW = Mae Tha, Lamphun; DM1C = Chiang Dao1, Chiang Mai; DM2C = Chiang Dao2, Chiang Mai; SM1C = San Pa Tong1, Chiang Mai; SM2C = San Pa Tong2, Chiang Mai; MPW = Mae Moh, Lampang. Data are expressed as mean ± standard deviation. The a–c values in the same column with different alphabets are statistically distinct (p > 0.05).
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

Thomya, S.; Wongkaew, M.; Bundithya, W.; Lumsangkul, C.; Jantrawut, P.; Sommano, S.R. Relationship between Phenotypes and Chemical Profiling of Boesenbergia rotunda Collected from Different Habitats of Northern Thailand. Agronomy 2023, 13, 1435. https://doi.org/10.3390/agronomy13061435

AMA Style

Thomya S, Wongkaew M, Bundithya W, Lumsangkul C, Jantrawut P, Sommano SR. Relationship between Phenotypes and Chemical Profiling of Boesenbergia rotunda Collected from Different Habitats of Northern Thailand. Agronomy. 2023; 13(6):1435. https://doi.org/10.3390/agronomy13061435

Chicago/Turabian Style

Thomya, Sureerat, Malaiporn Wongkaew, Weenun Bundithya, Chompunut Lumsangkul, Pensak Jantrawut, and Sarana Rose Sommano. 2023. "Relationship between Phenotypes and Chemical Profiling of Boesenbergia rotunda Collected from Different Habitats of Northern Thailand" Agronomy 13, no. 6: 1435. https://doi.org/10.3390/agronomy13061435

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