Next Article in Journal
Hydrogen Nanobubble Water Delays Petal Senescence and Prolongs the Vase Life of Cut Carnation (Dianthus caryophyllus L.) Flowers
Next Article in Special Issue
Plants Metabolome Study: Emerging Tools and Techniques
Previous Article in Journal
Photorhabdus spp.: An Overview of the Beneficial Aspects of Mutualistic Bacteria of Insecticidal Nematodes
Previous Article in Special Issue
A Novel Bioanalytical Method for Determination of Inotodiol Isolated from Inonotus Obliquus and Its Application to Pharmacokinetic Study
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Plants
Volume 10
Issue 8
10.3390/plants10081661
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

Metabolite Profiling and Dipeptidyl Peptidase IV Inhibitory Activity of Coreopsis Cultivars in Different Mutations

by
Bo-Ram Kim
1,2,†,
Sunil Babu Paudel
3,†,
Ah-Reum Han
1,†,
Jisu Park
1,
Yun-Seo Kil
3,
Hyukjae Choi
3,4,
Yeo Gyeong Jeon
5,
Kong Young Park
5,
Si-Yong Kang
6,
Chang Hyun Jin
1,
Jin-Baek Kim
1 and
Joo-Won Nam
3,*
1
Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute, Jeongeup-si 56212, Jeollabuk-do, Korea
2
Natural Product Research Division, Honam National Institute of Biological Resources, Mokpo-si 58762, Jeollanam-do, Korea
3
College of Pharmacy, Yeungnam University, Gyeongsan-si 38541, Gyeongsangbuk-do, Korea
4
Research Institute of Cell Culture, Yeungnam University, Gyeongsan 38541, Gyeongbuk, Korea
5
Uriseed Group, Icheon-si 17408, Gyeonggi-do, Korea
6
Department of Horticulture, College of Industrial Sciences, Kongju National University, Yesan-gun 32439, Chungcheongnam-do, Korea
*
Author to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Plants 2021, 10(8), 1661; https://0-doi-org.brum.beds.ac.uk/10.3390/plants10081661
Submission received: 20 July 2021 / Revised: 5 August 2021 / Accepted: 9 August 2021 / Published: 12 August 2021
(This article belongs to the Special Issue Isolation and Analysis of Characteristic Compounds from Herbal and Plant Extracts)
Browse Figures
Versions Notes

Abstract

:
Coreopsis species have been developed to produce cultivars of various floral colors and sizes and are also used in traditional medicine. To identify and evaluate mutant cultivars of C. rosea and C. verticillata, their phytochemical profiles were systematically characterized using ultra-performance liquid chromatography time-of-flight mass spectrometry, and their anti-diabetic effects were evaluated using the dipeptidyl peptidase (DPP)-IV inhibitor screening assay. Forty compounds were tentatively identified. This study is the first to provide comprehensive chemical information on the anti-diabetic effect of C. rosea and C. verticillata. All 32 methanol extracts of Coreopsis cultivars inhibited DPP-IV activity in a concentration-dependent manner (IC50 values: 34.01–158.83 μg/mL). Thirteen compounds presented as potential markers for distinction among the 32 Coreopsis cultivars via principal component analysis and orthogonal partial least squares discriminant analysis. Therefore, these bio-chemometric models can be useful in distinguishing cultivars as potential dietary supplements for functional plants.

1. Introduction

Coreopsis species are annual or perennial plants belonging to the Asteraceae (Compositae) family [1]. Approximately 80 species of Coreopsis are native to North America and are currently widespread in America, Asia, and Oceania regions [2,3,4]. They are usually cultivated for ornamental purposes in gardens or on roadsides. The plants are in the range of 46–120 cm in height and the petals of the flowers are primarily yellow in color and are serrated [5,6]. The color and size of Coreopsis flowers have commercially important value and are the reason for Coreopsis breeding. In addition, the Coreopsis flower has been ethnopharmacologically used for the treatment of diarrhea, vomiting, and hemorrhage in North America, where the Coreopsis species originates [7,8]. It has also been used as a drink to control diabetes in China and Portugal, and as an herbal tea to eliminate toxins and fever from the body in China [8, 9,10]. Nowadays, owing to scientific proof of its traditional use, several studies have been conducted on the phytochemical and biological activities of C. lanceolata and C. tinctoria, in particular [2,3,4,5,6,7,8,9,10,11,12,13,14,15,16,17,18,19]. Diverse types of flavonoids, such as aurone, chalcone, flavanone, and flavanol have been identified from C. lanceolata and C. tinctoria. In addition, unique polyacetylene compounds have also been found in these plants [10,12,13]. Various pharmacological activities such as anticancer [2,5], antioxidant [6,14,15,16,17,18], anti-inflammatory [6,10,19], and anti-diabetic effects [8,12] have been reported of compounds isolated from C. lanceolata and C. tinctoria. Limited reports on the chemical composition and biological activities of Coreopsis species exist.
In this study, several new cultivars of C. rosea and C. verticillata throughout γ-irradiated mutation or herbicide-induced artificial mutation were developed and registered in the Korea Seed and Variety Service (Table 1 and Figure 1) [20]. For horticultural purposes and the improvement of quality and functionality, numerous cultivars have been developed by hybridization, and mutations were induced by chemical mutagens and ionizing radiation in plant breeding programs [21]. Previous studies on C. rosea and C. verticillata have primarily focused on plant growth impact assessment, new variety development, and horticulture [22,23,24,25]. However, there has been no report on the phytochemical and biological activity of C. rosea and C. verticillata, except for our previous study on the volatiles’ composition and antioxidant activity of C. rosea cultivars [18].
As part of our investigation of the effects of mutation on metabolic changes between the original mutant cultivars and their biological functions, we analyzed metabolite profiling of the five original cultivars and each mutant cultivar. Given that Coreopsis species have been known to be effective for diabetes in folk medicine, 70% methanol extracts of 32 Coreopsis samples were evaluated for their inhibitory effect against dipeptidyl peptidase (DPP)-IV, a target of incretin-based therapies for the treatment of type 2 diabetes mellitus.

2. Results

2.1. Subsection Identification of Metabolites in Coreopsis Cultivars Using UPLC-QTof-MS

Metabolites in Coreopsis cultivars were tentatively identified using UPLC-QTof-MS. The Metabolites were separated with high resolution within 10 min in the base peak ion (BPI) chromatogram. BPI chromatograms of the original Coreopsis cultivars are shown in Figure S1. The mass spectrum of each peak was carefully interpreted by analyzing its experimental and theoretical high resolution MS (the deprotonated molecular ion, [M – H]−), error ppm, molecular formula, and MS/MS fragmentation. Additionally, these were compared with data from the literature of plants belonging to the same genus, such as C. tinctoria (known as snow chrysanthemum) and C. lanceolata [3,5,6,12,26,27,28,29]. Moreover, its mass spectrum was compared to that in Waters Traditional Medicine Library that is built in UNIFI software (Waters, Milford, MA, USA) and MassBank available online (a public database for sharing mass spectral data) [30,31]. Forty compounds, including phenolic acids, flavonoids, and a polyacetylene were identified in methanol extracts of original and mutant cultivars of C. rosea and C. verticillata (Table 2). However, a peak observed in total ion chromatograms of all, or some Coreopsis cultivars could not be identified in this study.

2.1.1. Phenolic Acids

Chlorogenic acid (peak 2, tR 4.50 min) produced a major molecular ion at m/z 353.0864 [M – H]− (calculated for C16H17O9−, 353.0878). At high energy scan, a fragment ion for a quinic acid was observed at m/z 191.0556 [3,26,27]. Peak 4 (tR 5.21 min) produced a molecular ion at m/z 329.0865 [M – H]− corresponding to C14H17O9− and produced two fragment ions at m/z 167.0338 and 151.0026, assigned to the loss of glucose and [M – H – glucose – O]−, respectively. This peak was tentatively identified as vanillic acid-4-glucoside, which was confirmed by the UNIFI local library and first detected in Coreopsis species. Peak 27 (tR 7.68 min) produced a major ion at m/z 515.1183 [M – H]− (calculated for C25H23O12−, 515.1195) with two stable fragment ions at m/z 353.0869 [M – H – C9H6O3]− and 191.0557 [M – H – 2C9H6O3]− assigned to dicaffeoylquinic acid. Peak 31 (tR 8.04 min) had the same precursor and fragment ions with peak 27. Peaks 27 and 31 were tentatively identified as 3,5-dicaffeoylquinic acid and 4,5-dicaffeoylquinic acid, respectively, by comparing with values in the literature of clearly identified constituents in C. tinctoria [26].

2.1.2. Flavanones and Flavanonols

It has been reported that a retro Diels–Alder reaction, as well as the loss of H2O, sugar (usually glucose), and carbonyl groups were observed in the ion fragmentation pathways of flavonoids [3]. Flavanones, chalcones, and their glycosides have been known as the major types of flavonoids found in Coreopsis species [28]. These compounds usually showed the loss of H2O caused by the disposition of hydroxyls at C-3′ and C-4′ in the flavanone structure or at C-3 and C-4 in the chalcone structure, and the loss of a glucose at C-7 in the flavone structure or at C-4′ in the chalcone structure [3]. These phenomena were observed in mass spectra of flavanones and chalcones identified in this study. Abundant fragment ions, [M – H – glucose]− and [M – H – glucose – H2O]− for their glycosides and [M – H – H2O]− for aglycones were produced by the loss of glucose and H2O, respectively. The loss of C8H6O (118 Da) or C8H6O2 (134 Da) were also characteristic fragment ions for flavanones and chalcones [3].
Peaks 1 (tR 4.48 min) and 3 (tR 4.96 min) produced a major molecular ion at m/z 465.1030 [M – H]− (calculated for C21H21O12−, 465.1038) and yielded fragment ions at m/z 303.0503 [M – H – glucose]− and 285.0397 [M – H – glucose – H2O]− by the loss of glucose and H2O, indicating the presence of a 3′,4′-dihydroxyphenyl group. The fragment ion at m/z 151.0034 [M – H – glucose – H2O – C8H6O2]− showed the presence of a 3-hydroxy group. Peak 3 had the fragment ion at 287.0550 [M – H – glucose – O]−, assuming that the sugar was attached at C-3 in the C ring. Therefore, peaks 1 and 3 were tentatively identified as taxifolin-7-O-glucoside [3] and taxifolin-3-O-glucoside [27], respectively. The aglycone of these compounds, taxifolin (peak 17, tR 6.87 min), showed the same fragment ions with those of its glycosides (peaks 1 and 3) [3,26]. Peak 5 (tR 5.74 min) produced a molecular ion at m/z 449.1085 [M – H]−, corresponding to the molecular formula C21H21O11−. At high energy scan, fragment ions at m/z 287.0554 [M – H – glucose]− and 269.0446 [M – H – glucose – H2O]− were detected by the loss of glucose and H2O and indicated the presence of a 3′,4′-dihydroxyphenyl group for the B ring. Fragment ions at m/z 151.0034 [M – H – glucose – H2O – C8H6O]− suggested no hydroxyl group at C-3. Thus, it was tentatively identified as flavanomarein [26]. The fragmentation pattern of peak 6 (tR 5.85 min) was identical to that of peak 5, except for the major molecular ion at m/z 595.1649 [M – H]− (calculated for C27H31O15−, 595.1668) and the fragment ion at m/z 449.1069 [M – H – rhamnose]. Therefore, peak 6 was predicted as isookanin-7-O-rutinoside, which has been described previously [27]. Given that peak 16 (tR 6.64 min) also exhibited the same fragment ions with those of peaks 5 and 6, it was tentatively identified as their aglycone, isookanin [3,26]. Peak 8 (tR 5.98 min) produced a molecular ion at m/z 433.1135 [M – H]− (calculated for C21H21O10−, 433.1140), which is 16 Da less than that of peak 5, and the fragment ion at m/z 135.0449 [M – H – glucose – H2O – C8H6O]− indicated the presence of a 3′,4′-dihydroxyphenyl group and no hydroxyl group at C-3 in the C ring, and consequently confirmed the presence of a hydroxyl group in the A ring. Thus, peak 8 was tentatively identified as butin-7-O-glucoside [3]. Peak 9 (tR 6.17 min) produced a molecular ion at m/z 479.0825 [M – H]−, corresponding to the molecular formula C21H19O13−. The fragment ions at m/z 317.0291 [M – H – glucose]− and 166.9963 [M – H – glucose – CH3 – H2O – C8H6O]− represented the presence of two hydroxy groups at C-3′ and C-4′ in the B ring and the absence of a hydroxy group at C-3 in the C ring, and consequently predicted the presence of 5,7-dihydroxy-8-methoxyphenyl group for the A ring. Therefore, peak 9 was tentatively identified as 8-methoxyeriodictyol-7-O-glucoside. Its aglycone, 8-methoxyeriodictyol, has been isolated from several plants [32,33,34]; however, its glycoside form has not been described previously. Peaks 10 (tR 6.23 min) and 11 (tR 6.34 min) produced the same molecular ion at m/z 463.1239 [M – H]− (calculated for C22H23O11−, 463.1246) and the same fragment ion at m/z 165.0188 [M – H – glucose – H2O – C8H6O]−, indicating the presence of a 3′,4′-dihydroxyphenyl group for the B ring and no hydroxyl group at C-3 in the C ring, and consequently predicted the presence of the 7-hydroxy-8-methoxyphenyl group for the A ring. Therefore, peaks 10 and 11 were tentatively identified as coreolanceoline B [12] and lanceolin [6], respectively. Peak 12 (tR 6.48 min) produced a molecular ion at m/z 433.1134 [M – H]− (calculated for C21H21O10−, 433.1140), exhibiting the same molecular ions as peak 8; however, an [M – H – glucose – C8H8O]− ion, instead of a [M – H – glucose – H2O]− ion of peak 8, was observed in the fragmentation pattern of peak 12, indicating the presence of one hydroxy group in the B ring and two hydroxy groups in the A ring. Accordingly, peak 12 was tentatively identified as naringenin-7-O-glucoside [26]. Peak 14 (tR 6.52 min) produced a molecular ion at m/z 595.1664 [M – H]− (calculated for C27H31O15−, 595.1668) and the fragment ions at m/z 433.1121 [M – H – glucose]− and 271.0604 [M – H – 2glucose]−, indicating the presence of two glucose groups. The fragment ion at m/z 135.0447 [M – H – 2glucose – C8H8O2]− without the loss of H2O resulted in the presence of two hydroxy groups at C-3′ and C-5′ in the B ring. Thus, peak 14 was tentatively identified as 7,3′,5′-trihydroxyflavanone-O-diglucoside. Given that 7,3′,5′-trihydroxyflavanone-7-O-glucoside and its aglycone have been found in Coreopsis species [3,28], three types of this compound, 7,3′,5′-trihydroxyflavanone-7-O-(glucosyl glucoside), 7,3′,5′-trihydroxyflavanone-7,3′-O-di glucoside, and 7,3′,5′-trihydroxyflavanone-7,5′-O-diglucoside were predicted as possible structures; however, the three compounds have not been described previously. Peak 18 (tR 6.91 min) produced a molecular ion at m/z 581.1501 [M – H]− (calculated for C26H29O15−, 581.1512) and the fragment ion at m/z 287.0552 [M – H – arabinose – glucose] by a loss of the arabinosyl-glucose. The fragment ion at m/z 167.0342 [M – H – arabinosyl-glucose – C8H6O] without the loss of H2O indicated the presence of a hydroxy group in the B ring and three hydroxyl groups in the A ring. Thus, peak 18 was tentatively identified as 4′,5,7,8-tetrahydroxyflavanone-7-O-(6-O-arabinosyl-glucoside), which has not been described previously. Its aglycone, isocarthamidin (4′,5,7,8-tetrahydroxyflavanone), has not been reported in Coreopsis species; however, it has been isolated from the Asteraceae plant [35]. Peak 24 (tR 7.26 min) produced a molecular ion at m/z 493.0984 [M – H]− (calculated for C22H21O13−, 493.0988) and fragment ions at 331.0447 [M – H – glucose]− and 316.0200 [M – H – glucose – CH3]− by loss of a glucose and a methyl of the methoxy group, respectively. The fragment ion at m/z 164.9830 [M – H – glucose – CH3 – H2O – C8H6O2]− produced a 3′-methoxy-4′-hydroxyphenyl group (or 3′-hydroxy-4′-methoxy phenyl group) for the B ring and a 3-hydroxy group in the C ring. Accordingly, peak 24 was tentatively identified as taxifolin 3′,7-dimethyl ether 3-O-glucoside [36]. Another candidate, taxifolin 4′,7-dimethyl ether 3-O-glucoside, has not been described previously; however, aglycones, taxifolin 4′,7-dimethyl ether, and taxifolin 3′,7-dimethyl ether have been reported in Asteraceae plants [37,38,39].

2.1.3. Chalcones

Peak 23 has the same fragment rules as flavanomarein; however, it has been known that flavanones have shorter retention times than chalcones in chromatographic elution [29]. Therefore, peak 23 (tR 7.15 min) was identified as marein [3,26,27]. The aglycone of this compound, okanin (peak 32, tR 8.26 min), produced identical fragment ions with peak 23 [3,26,27]. Similarly, peak 30 (tR 8.02 min) showed the same molecular ion and fragment ions as peak 8, thus identified as coreopsin [3,26]. Butein (peak 38, tR 9.20 min), the aglycone of peak 30, showed a molecular ion at m/z 271.0605 [M – H]− (calculated for C15H11O5−, 271.0612) and identical fragment ions with peak 30 [3,27]. Peak 13 (tR 6.51 min) produced a molecular ion at m/z 611.1612 [M – H]−, corresponding to the molecular formula C27H31O16−. At high energy scan, the fragment ions at m/z 449.1080, 287.0551, and 269.0393 were formed by the loss of one glucose, two glucoses, and H2O, respectively, indicating the presence of two glucose groups and a 3,4-dihydroxyphenyl group for the B ring. Hence, peak 13 was tentatively identified as okanin-3′,4′-O-diglucoside, which has been isolated from Bidens pilosa [40]. Peak 33 (tR 8.46 min) produced a molecular ion at m/z 611.1398 [M – H]−, corresponding to the molecular formula C30H27O14−. At high energy scan, fragment ions at m/z 449.1109 [M – H – glucose]− and 287.0559 [M – H – 2glucose]− were produced by the loss of two glucoses and 269.0441 [M – H – 2glucose – H2O]− by the loss of H2O, and the fragment ion at m/z 151.0024 [M – H – 2glucose – H2O – C8H6O]− indicated the presence of a 3′,4′-dihydroxyphenyl group for the B ring and the absence of a 3-hydroxy group in the C ring. Therefore, peak 33 was tentatively identified as eriodictyol chalcone-7-O-(glucosyl glucoside) or eriodictyol chalcone-O-diglucoside, which have not been described previously. Eriodictyol chalcone-7-O glucoside, which has one glucose, has been found in Antirrhinum majus [41]. Peak 34 (tR 8.73 min) produced a molecular ion at m/z 287.0553 [M – H]− and exhibited the same fragment pathway with that of peak 33, suggesting that it was an aglycone of peak 33, eriodictyol chalcone, which has been identified in Coreopsis species [42]. Peak 37 (tR 8.84 min) produced a molecular ion at m/z 477.1396 [M – H]−, corresponding to the molecular formula C23H25O11−. At high energy scan, fragment ions were produced at m/z 315.0864 [M – H –glucose]−, 300.0624 [M – H –glucose – CH3]−, 297.0754 [M – H –glucose – H2O]−, 282.0527 [M – H – glucose – H2O – CH3]−, 163.0747 [M – H – glucose – H2O – C8H6O]−, and 148.00524 [M – H – glucose – H2O – CH3 – C8H6O]−. Therefore, peak 37 was tentatively identified as 4-methoxylanceoletin-4′-O-glucoside, which has been isolated from C. lanceolata [12], or lanceolein 2′-methyl ether, which has not been described previously.

2.1.4. Flavones and Flavanols

Flavone having a double bond between C-2 and C-3 exhibits a molecular ion that is 2 Da less than that of flavanone or chalcone and characteristic fragment ions by the loss of C8H4O (116 Da) or C8H4O2 (132 Da) [3]. Peak 7 (tR 5.93 min) produced a molecular ion at m/z 609.1454 [M – H]− (calculated for C27H29O16−, 609.1461) and fragment ions at m/z 447.0932 [M – H – glucose]− and 285.0392 [M – H – glucose – glucose]− by the loss of two glucoses. Another fragment ion at m/z 151.0033 [M – H – 2glucose – H2O – C8H4O]− indicated the presence of a 3′,4′-dihydroxyphenyl group for the B ring without a 3-hydroxy group in the C ring. Thus, peak 7 was tentatively identified as luteolin-7-O-sophoroside [3]. Peaks 22 (tR 7.03 min) and 36 (tR 8.80 min) produced molecular ions at m/z 447.0929 [M – H]− (calculated for C21H19O11−, 447.0927) and m/z 285.0398 [M – H]− (calculated for C15H9O6−, 285.0405) that were 162 Da and 324 Da less than that of peak 8, respectively, indicating that the sugar moiety was removed from C-2′′ and C-7 in luteolin-7-O-sophoroside (peak 7). Accordingly, peaks 22 and 36 were tentatively identified as luteolin-7-O-glucoside and luteolin, respectively [3,26,27]. In addition, a molecular ion at m/z 431.0978 [M – H]− (calculated for C21H19O10−, 431.0984) for peak 29 (tR 7.91 min) was 146 Da more than that of peak 36, indicating the addition of a rhamnose. Moreover, its fragment ions were similar to those of peaks 22 and 36. Thus, it was identified as luteolin-7-O-rhamnoside, which was first detected in Coreopsis species; however, it has been found in other plants, such as Glechoma grandis Kuprianova var. longituba, Rumex algeriensis, and Cornulaca monacantha [43,44,45]. Peak 15 (tR 6.58 min) produced a molecular ion at m/z 609.1454 [M – H]− (calculated for C27H29O16−, 609.1461) and fragment ions at m/z 447.0932 [M – H – glucose]− and 285.0394 [M – H – 2glucose]− by the loss of two glucoses. The fragment ion at m/z 151.0033 [M – H – 2glucose – H2O – C8H4O2]− suggested the presence of a 3′,4′-dihydroxyphenyl group for the B ring and a 3-hydroxy group in the C ring. As a result, peak 15 was tentatively identified as fisetin-3,7-O-diglucoside, which was first detected in Coreopsis species; however, it has been found in other Sophora species [46]. Other glycosides of fisetin have not been previously described. Peak 20 (tR 6.97 min) produced a molecular ion at 463.0885 [M – H]−, corresponding to the molecular formula C21H19O12−. At a high energy scan, the fragment ion at m/z 301.0346 [M – H – glucose]− was observed by the loss of a glucose, and fragment ions at m/z 151.0034 [M – H – glucose – H2O – C8H4O2]− indicated the presence of a 3′,4′-dihydroxyphenyl group by the loss of H2O and the presence of a 3-hydroxy group. Therefore, peak 20 was tentatively identified as quercetin-7-O-glucoside [3,26]. Peak 25 (tR 7.33 min) showed a molecular ion at m/z 461.1085 [M – H]− (calculated for C22H21O11−, 461.1089). At high energy scan, fragment ions at m/z 299.0547 [M – H – glucose]−, 283.0242 [M – H – glucose – O]−, 165.0188 [M – H – glucose – O – H2O – C8H4O]−, and 133.0291 [M – H – glucose – O – C8H6O3]− were observed, indicating the loss of a glucoside at a 3-hydroxy group in the C ring and the presence of 3′,4′-dihydroxyphenyl group for the B ring. Therefore, peak 25 was tentatively identified as 3,3′,4′-trihydroxy-7-methoxyflavone 3-O-glucoside, which has been reported in Aptenia cordifolia [47]. Peak 26 (tR 7.58 min) produced a molecular ion at m/z 641.1141 [M – H]−, corresponding to the molecular formula C30H25O16−. At high energy scan, fragment ions were produced at m/z 317.0294 [M – H – caffeoylglucose]−, 301.0342 [M – H – caffeoylglucose – O]−, 285.0381 [M – H – caffeoylglucose – O – O]−, 179.0343 [M – H – C15H9O8 – C6H10O4]−, 161.0224 [M – H – C15H9O8 – C6H10O4 – O]−, 135.0447 [M – H – C15H9O8 – C6H10O4 – CO2]−, and 133.0289 M – H – caffeoylglucose – O – O – C7H3O4]−. This peak was tentatively identified as qurcetagetin-7-O-(6′′-caffeoylglucoside), which was confirmed using the UNIFI local library and was first detected in Coreopsis species. However, this compound has been found in Asteraceae plants, such as Gnaphalium uliginosum and Tagetes maxima [48,49]. Peak 35 (tR 8.74 min) produced a molecular ion at m/z 299.0555 [M – H]− (calculated for C16H11O6−, 299.0561). The fragment ions produced at m/z 284.0319 [M – H – CH3]− by the loss of a methyl of a methoxy group were observed. Moreover, 151.0032 [M – H – CH3 – C8H5O2]− indicated the presence of a hydroxyl group at the B ring and a 3-hydroxy group in the C-ring. Other fragment ions at m/z 151.0032 [M – H – CH3 – C8H6O2]− and 133.0447 [M – H – CH3 – C7H3O4]− indicated the presence of a 4′-hydroxyphenyl group for the B ring with a 3-hydroxy group and two hydroxyl groups in the A ring, respectively. Thus, peak 35 was tentatively identified as kaempferide, by comparison of its mass spectrum with that in the MassBank database [31]. This compound was first detected in Coreopsis species; however, it has been found in Asteraceae plants, such as Chrysanthemum morifolium, C. coronarium, Artemisia annua, Chromolaena odorata, and Filago germanica [50,51,52]. Peak 39 (tR 9.28 min) produced a molecular ion at m/z 269.0447 [M – H]− (calculated for C15H9O5−, 269.0450) and fragment ions at m/z 227.0351 [M – H– C2H2O]− and 117.0341 [M – H– C7H4O4]−. Thus, it was tentatively identified as apigenin [26].

2.1.5. Aurones

Aurones are also one of the characteristic flavonoids found in Coreopsis species [3,12,29]. Peak 19 (tR 6.94 min) produced a molecular ion at m/z 431.0977 [M – H]− (calculated for C21H19O10−, 431.0984) and a fragment ion at m/z 269.0447 [M – H – glucose]− by the loss of a glucose. In addition, fragment ions at m/z 135.0447 [M – H – glucose – H2O – C8H4O]− and 133.0447 [M – H – glucose – C7H4O3]− were also observed, indicating the presence of a 3′,4′-dihydroxyphenyl group for the B ring. Thus, peak 19 was tentatively identified as sulfurein (sulfuretin-6-O-glucoside) [35,53]. Sulfuretin (peak 28, tR 7.77 min), the aglycone of peak 19, showed a molecular ion at m/z 269.0449 [M – H]− (calculated for C15H9O5−, 269.0450) and identical fragment ions with peak 19 [27]. Peak 21 (tR 7.01 min) showed a molecular ion at m/z 447.0929 [M – H]− (calculated for C21H19O11−, 447.0927), which is 16 Da more than that of peak 20, and fragment ions at m/z 285.0397 [M – H – glucose]−, 135.0447 [M – H – glucose – H2O – C8H4O – O]−, and 133.0447 [M – H – glucose – C7H4O3]−, thus, identified as maritimein [3]. This identification was also confirmed by comparison with ESI-QTof-MS (negative ion mode) of maritimein in the MassBank database [31].

2.1.6. Polyacetylene

Polyacetylenes of various structures have been isolated from the genus Coreopsis [12,28]. In this study, peak 41 (tR 9.45 min) produced a molecular ion at m/z 557.2219 [M – H]−, a molecular formula of C26H37O13−. At high energy scan, fragment ions were produced at m/z 233.0650 [M – H – 2glucose]−, 191.0554 [M – H – 2glucose – C3H6]−, and 149.0441 [M – H – caffeoylglucose – C5H8O]−. This peak was tentatively identified as lobetyolinin, which was confirmed by the UNIFI local library and first detected in Coreopsis species; however, it has primarily been found in Lobelia species [54,55].

2.2. DPP-IV Inhibitory Effects of the 70% Ethanol Extract Obtained from Coreopsis cultivars

Type 2 diabetes mellitus is determined by several factors, including pancreas β-cell dysfunction, insulin resistance, increased hepatic and intestinal glucose production, or deficient insulin secretion [56]. Recently, the incretin effect has been observed to be reduced in patients with type 2 diabetes mellitus, which is a symptom of increased insulin secretion induced by oral administration, such as eating a meal, compared to intravenous administration of glucose [56]. This effect is mediated by incretin hormones, glucagon-like peptide-1 (GLP-1), and glucose-dependent insulinotropic polypeptide (GIP), which stimulate insulin secretion from pancreatic β-cells and consequently increase the blood glucose level [57,58]. In the incretin system, an increase of the elimination of GLP-1 and GIP occurs primarily through enzymatic degradation of DPP-IV [59]. Thus, DPP-IV inhibition enhances the function of insulinotropic hormones. It improves glucose tolerance in patients with type 2 diabetes mellitus [58]. Hence, DPP-IV inhibitors have emerged as a new class of oral anti-diabetic agents, and synthetic compounds have mainly been used in current treatments with these inhibitors [59]. However, there have also been studies that show that DPP-inhibitors are derived from natural sources as promising candidates of functional foods or pharmaceuticals [12,60,61,62,63].
In this study, the 70% ethanol extract of original and mutant cultivars of C. rosea and C. verticillata confirmed their anti-diabetic effect using an in vitro DPP-IV inhibitor screening assay. All extracts inhibited DPP-IV activity in a concentration-dependent manner with IC50 values from 34.01 to 134.28 μg/mL (Table 3). The positive control, sitagliptin, exhibited an IC50 of 0.095 μM. In two different species, the cultivars of C. rosea (Groups I and III) showed less inhibition of DPP-IV than the cultivars of C. verticillata (Groups II, IV, and V). Of the 32 samples, ‘Orange sunlight (No. 30)’, which belongs to Group IV (C. verticillata ), showed the greatest DPP-IV inhibitory effects. Thereafter, the most active cultivars with IC50 values less than 65 μg/mL were in the order of ‘Golden sunlight (No. 26)’, ‘Golden ball No.48 (No. 18)’, ‘Golden ball No.42 (No. 17)’, ‘Red sunlight (No. 27)’, ‘Bright sunlight (No. 28)’, and ‘Golden ball No.21 (No. 15)’, all of which belonged to C. verticillata. The DPP-IV inhibitory effects of six mutant cultivars, ‘Lemon candy (No. 4)’, ‘Shiny pink (No. 5)’, ‘Uri-dream 01 (No. 6)’, ‘Luckyten5 (No. 7)’, ‘Luckyten9 (No. 8)’, and ‘Uri-dream red (No. 9)’ were greater by 24–47 % than that of the original cultivar, ‘Heaven’s gate (No. 1)’ in Group I, while other mutant cultivars, ‘Luckyten 6 (No. 2)’, ‘Redfin (No. 3)’, ‘Uri-dream 07 (No. 10)’, ‘Uri-dream 06 (No. 11)’, and ‘Pink sherbet (No. 12)’ had similar or lower efficacy. In Group II, except for ‘Golden ball No.26 (N. 16)’, four mutant cultivars, ‘Golden ball No.18 (No. 14)’, ‘Golden ball No.21 (No. 15)’, ‘Golden ball No.42 (No. 17)’, and ‘Golden ball No.48 (No. 18)’ showed a 5–26 % increase in the inhibitory effect of DPP-IV compared to the original cultivar, ‘Citrine (No. 9)’. In Groups III and V, mutant cultivars exhibited similar or lower DPP-IV inhibitory effects than original cultivars. In Group IV, compared to the original cultivar, ‘Route 66 (No. 25)’, all mutant cultivars presented 7–37 % higher inhibitory effects of DPP-IV.
Among all Coreopsis cultivars samples, ‘Orange sunlight (No. 30)’ showed the best efficacy with an IC50 value of 34.01 μg/mL; however, ‘Uri-dream red (No. 9)’ (IC50, 66.46 μg/mL) had the highest increase with 47% DPP-IV inhibitory activity compared to the original cultivar (IC50, 125.29 μg/mL). Therefore, ‘Orange sunlight (No. 30)’ had the potential to develop as a functional food, such as a tea ingredient or a food additive for the prevention or treatment of type 2 diabetes. The mutant cultivars with a greater increase in activity compared to the original cultivar, such as ‘Uri-dream red (No. 9)’ may be used for studies to identify metabolites changed by mutation using multivariate analysis, and for further research on genomic mutation mechanism.

2.3. Multivariate Analysis

Metabolite differences among original and mutant cultivars, C. rosea and C. verticillate, were examined based on the metabolite profiles analyzed by UPLC-QTof-MS. However, it was difficult to find differences among the samples in chromatograms. Therefore, PCA and OPLS-DA were used to provide an effective visualization for the classification and differentiation of a metabolome system.
To compare metabolites from different cultivars of C. rosea and C. verticillata, we performed PCA analysis on negative ion mode data obtained from UPLC-QTof-MS analysis. PCA analysis was performed with three principal components (PC1–PC3) describing variation explained, 0.66 of R2X and predictive capability, 0.366 of Q2. Eigenvalues for PC1 and PC2 were found to be 9.94 and 8.05, respectively, indicating these first two principal components explain a large amount of the variance in the data. PC3 showed a comparatively smaller eigenvalue of 3.13, which led us to choose only PC1 and PC2 for further analysis. As shown in Figure 2A, the first two principal components described 56.2% of the total variation (31.1% and 25.1% by PC1 and PC2, respectively), and 32 Coreopsis samples were clearly clustered into four groups. Group I and Group III were clustered together, indicating similar chemical profiles among samples, and these two groups were of the same species, C. rosea. This cluster also suggested that there is no distinct difference between original cultivars and other mutation cultivars induced from each original one. However, an exception was found in ‘Luckyten 6 (No. 2)’, which is one of the mutant cultivars artificially induced using herbicide from the original cultivar of Group I, ‘Heaven’s gate (No. 1)’. Alternatively, a distinct separation was observed from cultivars in Group II, Group IV, and Group V, although they were all included in C. verticallata. Group II demonstrated different chemical profiles compared with Group IV and Group V. However, Group II showed one clustering with no substantial deviations between the γ-irradiated mutant cultivars (No.14–No. 18) and the original cultivar ‘Citrine (No. 13)’. In Group IV, the γ-irradiated mutant cultivar, ‘Orange sunlight (No. 30)’ was shown as the outlier, indicating that it had a different chemical profile than samples within the same group. Figure 2B, shows the derivation of markers primarily distributed among the four groups. However, this resulted in whole variability directions, with no distinction of variabilities among groups. Accordingly, we performed OPLS-DA analysis on the metabolite profiles between C. rosea cultivars (Group Cr) and C. verticallata cultivars (Group Cv) to find the differentiation and significant variances in these two species. Two clusters were clearly differentiated from each other according to species in the OPLS-DA model, with a cumulative R2Y value of 1.00 and a cumulative Q2 value of 0.94 (Figure 2C). However, ‘Luckyten 6 (No. 2)’ and ‘Orange sunlight (No. 30)’ were marginally out of each grouped sample area. The internal validation of OPLS-DA model was performed by a permutation test (n = 200). In permutation test, the intercept values of R2 and Q2 were 0.425 and −1.09 respectively. All permutations of the R2 and Q2 values to the left were lower than the original points to the right and the intersection of regression lines of the R2 and Q2 points on vertical axis was below 0.4 and −1.1, respectively (Figure S43). These values indicated OPLS-DA model of this analysis was strongly validated without overfitting of the original model. As shown in Figure 2D, the corresponding OPLS-DA S-plot enabled the derivation of 13 potential marker compounds responsible for separating two groups by being far from the center. Eight marker metabolites which were shifted in the same direction as Group Cv from the OPLS-DA score plot were peaks 2, 8, 19, 21, 27, 30, 36, and 38, indicating the most abundant markers in Group Cv. Five marker metabolites, peaks 1, 5, 20, 23, and 34, were at the highest level in Group Cr. The variable importance plot (VIP) (Figure 2E) confirms these 13 selected marker compounds are primarily responsible for the discrimination between Group Cr and Group Cv with high VIP values (VIP ≥ 1). Moreover, the variable average by group clearly shows differences of selected marker compounds (Figure 2F) in these groups.
The similarities in chemical composition and relative quantitative differences among different cultivars of C. rosea and C. verticallata were clearly visualized on a heatmap with a dendrogram, while a hierarchical cluster analysis exhibited the same pattern of clustering as observed in PCA analysis (Figure 3). Heatmap is considered as one of the best tools for converting qualitative data into quantitative. Group I (No. 1–No. 12) and Group III (No. 19–No. 24) were clustered as one big cluster with similar distribution of areas of peaks 1, 3, 4, 5, 9, 11, 12, 13, 17, 20, 21, 22, 23, 24, 26, 27, 34, 40, and 41. ‘Luckyten 6’ (No. 2) was observed to have comparatively higher area values for peaks 1, 3, 20, and 34 than other cultivars in Group I, indicating the relatively high contents of these four peaks when compared to other samples in Group I. These four peaks could be responsible for making ‘Luckyten 6’ (No. 2) an outlier. Peaks 6, 25, 31, 35, and 37 appear with intense color in heatmap representing high quantity in comparison with other samples, which was responsible for the clustering of group II (No. 13–No. 18). Group IV has peaks 2, 8, 10, 14, 19, 28, 30, and 36 in abundance, while ‘Orange sunlight’ (No. 30) is rich in peaks 8, 10, 14, 19, 28, and 30 among groups. These six peaks’ composition and relatively higher content could turn ‘Orange sunlight’ (No. 30) into an outlier in this statistical study. The contents of eight peaks 7, 14, 15, 29, 32, 33, 36, and 39 determine the clustering of group V (No. 31–No. 32), adjacent to group IV, sharing some similarities between them.
The results of multivariate analyses to verify the correlation between metabolites and DPP-IV activities of the 32 Coreopsis samples were similar to the chemometric patterns between the two species. Given that DPP-IV inhibitory activities of C. verticallata cultivars appeared greater than that of C. rosea cultivars, distinguished metabolites between the active and inactive groups were almost identical to metabolites that showed differences between the two Coreopsis species presented in Table 3 (C. verticallata: IC50 < 65 µg/mL and C. rosea: IC50 > 65 µg/mL). Notably, ‘Orange sunlight (No. 30)’ and ‘Luckyten 6 (No. 2)’, which are outliers of Group Cv and Group Cr, respectively, were found to have the greatest and lowest DPP-IV inhibitory activity, respectively. These results suggested that the composition and relative content of distinguishable markers between C. rosea and C. verticallata cultivars were evaluated as key markers for the classification of species and contribution to the correlation of active and inactive cultivars.

3. Materials and Methods

3.1. Plant Material

Coreopsis cultivars were grown and collected from a wild cultivation field at Uriseed Group, Icheon-si, Gyeonggi-do, Republic of Korea and authenticated by Yeo Gyeong Jeon and Kong Young Park. These cultivars were selected according to their diverse phenotypic variants and exhibited a stable inheritance of these phenotypes for 4 years. Among them, five γ-irradiated mutants (Redfin, Lemone candy, Shiny pink, Uri-dream red, pink sherbet) of the original cultivar (Heaven’s gate) and the series of γ-irradiated mutants of original cultivars (Citirne, Pumpkin pie, Route 66) were generated using γ (60Co) irradiation (150 TBq capacity; AECL, Ottawa, ON, Canada). Six other mutant cultivars of ‘Heaven’s gate’ (Luckyten 6, Uri-dream 01, Luckyten5, Luckyten9, Uri-dream 07, Uri-dream 06) were artificially mutated using an herbicide. ‘Moonlight sonata’ was selected as the phenotypic variation of the original cultivar ‘Moonbeam’. Flowers used in this study were handpicked at the flowering stage in August 2018. These flowers were freeze-dried and stored at −20 °C for further analysis. Voucher specimens were deposited at the Uriseed Group Corporation.

3.2. Sample Preparation

Freeze-dried flowers of Coreopsis cultivars were ground into powder using a mixer. Extractions were performed with 200 mg of this powder in 20 mL of 70% methanol using an ultrasonic bath for 60 min, and subsequently evaporated to achieve a dry product. Thereafter, these dried extracts (1 mg each) were dissolved in 1 mL of 70% methanol and filtered through a 0.20 μm polyvinylidene fluoride filter. Samples (1000 ppm) were diluted with 70% methanol to a concentration of 200 ppm for further liquid chromatography–mass spectrometry (LC-MS) analysis. For the evaluation of bioactivity, methanol extracts were initially dissolved in dimethyl sulfoxide (DMSO) at a concentration of 10 mg/mL stock solution. All extraction and chromatographic solvents used in this study were of analytical grade (J. T. Baker, Phillipsburg, NJ, USA).

3.3. Ultra-Performance Liquid Chromatography Time-of-Flight Mass Spectrometry (UPLC-QTof-MS) Analysis

A Waters ACQUITY UPLC I-Class system equipped with a binary solvent pump and an autosampler combined with a Xevo G2-XS QTof-MS (Waters, Milford, MA, USA) was used. Each sample (1 μL) was injected into a ACQUITY UPLC BEH C18 column (2.1 mm i.d. × 100 mm, 1.7 μm) at a flow rate of 0.5 mL/min. The temperature of the column oven was maintained at 15 °C. The mobile phase was composed of 0.1% formic acid in water (v/v; solvent A) and 0.1% formic acid in acetonitrile (v/v; solvent B). Gradient elution was carried out as follows: 0–1.0 min, 1% B; 1.0–7.0 min, 1–20% B; 7.0–11.0 min, 20–40% B; 11.0–14.0 min, 60–100% B; 14.0–14.5 min, 100% B; 14.5–15.0 min, 100–1% B and 15.0–17.0 min, 1% B. The mass spectrometer was operated in negative ion mode with the following parameters: source temperature, 120 °C; desolvation temperature, 400 °C; capillary voltage, 3.0 kV; cone voltage, 40 V; cone gas flow: 50 L/h; flow rate of desolvation gas (N2), 1000 L/h; mass scan range, 550–1500 Da; scan time, 0.1 s. Leucine-enkephalin was used for the lock mass ([M ‒ H]− m/z 554.2615). Full scan data, MS/MS spectra, accurate mass, and elemental composition were calculated using UNIFI software (Waters, Milford, MA, USA).

3.4. DPP-IV Inhibitor Screening Assay

DPP-IV activity of Coreopsis cultivars was analyzed using a DPP-IV inhibitor screening assay kit (Cayman Chemical, Ann Arbor, MI, USA) which provides a fluorescence-based method for screening DPP-IV inhibitors. The assay uses the fluorogenic substrate, Gly-Pro-Aminomethylcoumarine (AMC), to measure DPP-IV activity. Cleavage of the peptide bond by DPP releases the free AMC group, resulting in fluorescence that can be analyzed using an excitation wavelength of 350–360 nm and an emission wavelength of 450–465 nm. The tested samples dissolved in DMSO at a concentration of 10 mg/mL were subsequently diluted to a final concentration of 20 to 200 μg/mL using DMSO and were added to a 96-well plate to a final volume of 10 μL and a final concentration of 50 μM. The assay procedure is described in our previous studies [12,62,63]. Briefly, diluted assay buffer (30 μL) and diluted enzyme solution (10 μL) were added to the 96-well plate containing 10 μL of solvent (blank) or solvent-dissolved test samples. The reaction was initiated by adding 50 μL of a diluted substrate solution, and the plate was incubated for 30 min at 37 °C. Following incubation, fluorescence with an excitation wavelength of 350 nm and an emission wavelength of 450 nm was monitored using a plate reader (TECAN, Männedorf, Switzerland). The percent inhibition was expressed as ([DPP-IV level of vehicle-treated control − DPP-IV level of test samples]/DPP-IV level of vehicle-treated control) × 100. Subsequently, the 50% inhibitory concentration (IC50) was determined using GraphPad Prism software (GaraphPad Software, La Jolla, CA, USA) via dose–response analysis.

3.5. Chemometric Data Analysis

Data management for the UPLC-QTof-MS analysis was performed using UNIFI software (Waters, Milford, MA, USA). MS data were processed using UNIFI to obtain a data matrix containing retention times, accurate masses, and normalized peak intensities. Parameters included retention time (tR, range of 0.0–15.0 min), mass-to-charge ratio (m/z, range of 100–1500 Da), and a mass tolerance of 0.04 Da. The resulting data were evaluated using SIMCA 15.0.2 (Umetrics, Umeå, Sweden) for multivariate statistical analysis. Unsupervised principal component analysis (PCA) was performed using UV (univariate)-scaled and supervised orthogonal partial least-squares discriminant analysis (OPLS-DA) was used to identify and compare different metabolite sizes of the 32 samples. The quality of the OPLS-DA model was evaluated with R2Y value and cumulative Q2 value. The model was further validated with a permutation test (n = 200). Markers for the difference between groups were identified by analyzing the S-plot with pareto scaling, which were generated with covariance (p) and correlation (pcorr) data. These data sets were normalized by dividing with mean value to get a value between 0 and 10 and a heatmap with dendrograms was generated using OriginPro 2021 (OriginLab Corporation, Northampton, MA, USA) selecting ward for cluster method. Marker compounds were tentatively identified by comparison to published MS data in literature and databases such as Waters Local Library in UNIFI and Massbank [3,5,6,26,27,28,29,30,31].

4. Conclusions

To the best of our knowledge, a comparative metabolomics approach to identify metabolite composition and DPP-IV inhibitory activities in various cultivars of C. rosea and C. verticillata, were demonstrated for the first time in this study. UPLC-QTof-MS techniques were used to identify several phenolic acids, flavonoids and a polyacetylene in mutant cultivars compared to original cultivars. PCA and OPLS-DA results showed that metabolites discriminate between the mutant and original cultivars and between the two species. In addition, significant changes in metabolite content were observed under different DPP-IV inhibitory activities of cultivars, and chlorogenic acid, butin-7-O-glucoside, sulfuretin-6-O-glucoside, maritimein, 3,5-dicaffeoylquinic acid, coreopsin, luteolin, and butein were abundant in the active extracts. Therefore, the DPP-IV inhibitory cultivars and the metabolites influencing their activities would be favorable for the development of functional foods and the information of the metabolites accumulated differently for each mutant cultivar would be useful as a scientific reference for further studies on plant mutation mechanisms.

Supplementary Materials

The following are available online at https://0-www-mdpi-com.brum.beds.ac.uk/article/10.3390/plants10081661/s1, Figure S1: title, Table S1: title, Video S1: title. Figure S1: Representative UPLC-QTof-MS chromatograms of the 70% methanol extracts of the original cultivars at low CE scan (6 eV) for precursor (up) and high CE scan (20-50 eV) for fragment ions (down). (a) ‘Heaven’s gate’ (No. 1), (b) ‘Citrine’ (No. 13), (c) ‘Pumpkin Pie’ (No. 19), (d) ‘Route 66’ (No. 25), and (e) ‘Moonbeam’ (No. 31), Figure S2: ESI-QTof-MS spectrum of taxifolin-7-O-glucoside (peak 1), Figure S3: ESI-QTof-MS spectrum of chlorogenic acid (peak 2), Figure S4: ESI-QTof-MS spectrum of taxifolin-3-O-glucoside (peak 3), Figure S5: ESI-QTof-MS spectrum of vanillic acid-4-glucoside (peak 4), Figure S6: ESI-QTof-MS spectrum of flavanomarein (peak 5), Figure S7: ESI-QTof-MS spectrum of isookanin-7-O-rutinoside (peak 6), Figure S8: ESI-QTof-MS spectrum of luteolin-7-O-sophoroside (peak 7), Figure S9: ESI-QTof-MS spectrum of butin-7-O-glucoside (peak 8), Figure S10: ESI-QTof-MS spectrum of 8-methoxyeriodictyol-7-O-glucoside (peak 9), Figure S11: ESI-QTof-MS spectrum of coreolanceoline B (peak 10), Figure S12: ESI-QTof-MS spectrum of lanceolin (peak 11), Figure S13: ESI-QTof-MS spectrum of naringenin-7-O-glucoside (peak 12), Figure S14: ESI-QTof-MS spectrum of okanin-3′,4′-O-diglucoside (peak 13), Figure S15: ESI-QTof-MS spectrum of 4′,7,8-trihydroxyflavone-O-diglucoside (peak 14), Figure S16: ESI-QTof-MS spectrum of fisetin-3,7-O-diglucoside (peak 15), Figure S17: ESI-QTof-MS spectrum of isookanin (peak 16), Figure S18: ESI-QTof-MS spectrum of taxifolin (peak 17), Figure S19: ESI-QTof-MS spectrum of 4′,5,7,8-tetrahydroxyflavanone-7-O-(6-O-arabinosyl-glucoside) (peak 18), Figure S20: ESI-QTof-MS spectrum of sulfuretin-6-O-glucoside (peak 19), Figure S21: ESI-QTof-MS spectrum of quercetin-7-O-glucoside (peak 20), Figure S22: ESI-QTof-MS spectrum of maritimein (peak 21), Figure S23: ESI-QTof-MS spectrum of luteolin-7-O-glucoside (peak 22), Figure S24: ESI-QTof-MS spectrum of marein (peak 23), Figure S25: ESI-QTof-MS spectrum of taxifolin 3′,7-dimethyl ether 3-O-glucoside (peak 24), Figure S26: ESI-QTof-MS spectrum of 3,3′,4′-trihydroxy-7-methoxyflavone 3-O-glucoside (peak 25), Figure S27: ESI-QTof-MS spectrum of qurcetagetin-7-O-(6′′-caffeoylglucoside) (peak 26), Figure S28: ESI-QTof-MS spectrum of 3,5-dicaffeoylquinic acid (peak 27), Figure S29: ESI-QTof-MS spectrum of sulfuretin (peak 28), Figure S30: ESI-QTof-MS spectrum of luteolin-6-O-rhamnoside (peak 29), Figure S31: ESI-QTof-MS spectrum of coreopsin (peak 30), Figure S32: ESI-QTof-MS spectrum of 4,5-dicaffeoylquinic acid (peak 31), Figure S33: ESI-QTof-MS spectrum of okanin (peak 32), Figure S34: ESI-QTof-MS spectrum of eriodictyol chalcone-O-diglucoside (peak 33), Figure S35: ESI-QTof-MS spectrum of eriodictyol chalcone (peak 34), Figure S36: ESI-QTof-MS spectrum of kaempferide (peak 35), Figure S37: ESI-QTof-MS spectrum of luteolin (peak 36), Figure S38: ESI-QTof-MS spectrum of 4-methoxylanceoletin-4′-O-glucoside (peak 37), Figure S39: ESI-QTof-MS spectrum of butein (peak 38), Figure S40: ESI-QTof-MS spectrum of apigenin (peak 39), Figure S41: ESI-QTof-MS spectrum of unknown (peak 40), Figure S42: ESI-QTof-MS spectrum of lobetyolinin (peak 41), Figure S43: Validation plot of the OPLS-DA obtained from 200 permutation test.

Author Contributions

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

Funding

This research was supported by Radiation Technology R&D program (No. 2017M2A2A6A05018541) through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT & Future Planning.

Acknowledgments

The authors thank Waters Korea (Seoul, Republic of Korea) for technical support for UPLC-QTof-MS experiments.

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. Kim, S.-C.; Crawford, D.J.; Tadesse, M.; Berbee, M.; Ganders, F.R.; Pirseyedi, M.; Esselman, E.J. ITS sequences and phylogenetic relationships in Bidens and Coreopsis (Asteraceae). Syst. Bot. 1999, 24, 480–493. [Google Scholar] [CrossRef]
  2. Pardede, A.; Mashita, K.; Ninomiya, M.; Tanaka, K.; Koketsu, M. Flavonoid profile and antileukemic activity of Coreopsis lanceolata flowers. Bioorg. Med. Chem. Lett. 2016, 26, 2784–2787. [Google Scholar] [CrossRef] [PubMed]
  3. Yang, Y.; Sun, X.; Liu, J.; Kang, L.; Chen, S.; Ma, B.; Guo, B. Quantitative and qualitative analysis of flavonoids and phenolic acids in snow chrysanthemum (Coreopsis tinctoria Nutt.) by HPLC-DAD and UPLC-ESI-QTOF-MS. Molecules 2016, 21, 1307. [Google Scholar] [CrossRef] [Green Version]
  4. Nakabo, D.; Okano, Y.; Kandori, N.; Satahira, T.; Kataoka, N.; Akamatsu, J.; Okada, Y. Convenient synthesis and physiological activities of flavonoids in Coreopsis lanceolata L. Petals and their related compounds. Molecules 2018, 23, 1671. [Google Scholar] [CrossRef] [Green Version]
  5. Kim, H.-G.; Oh, H.-J.; Ko, J.-H.; Song, H.S.; Lee, Y.-G.; Kang, S.C.; Lee, D.Y.; Baek, N.-I. Lanceolein A–G, hydroxychalcones, from the flowers of Coreopsis lanceolate and their chemopreventive effects against human colon cancer cells. Bioor. Chem. 2019, 85, 274–281. [Google Scholar] [CrossRef]
  6. Kim, H.-G.; Jung, Y.S.; Oh, S.M.; Oh, H.-J.; Ko, J.-H.; Kim, D.-O.; Kang, S.C.; Lee, Y.-G.; Baek, N.-I. Coreolanceolins A–E, new flavanones from the flowers of Coreopsis lanceolata, and their antioxidant and anti-inflammatory effects. Antioxidants 2020, 9, 539. [Google Scholar] [CrossRef] [PubMed]
  7. Gaspar, L.; Oliveira, A.P.; Silva, L.R.; Andrade, P.B.; Pinho, P.G.D.; Botelho, J.; Valentão, P. Metabolic and biological prospecting of Coreopsis tinctoria. Rev. Bras. Farmacogn. 2012, 22, 350–358. [Google Scholar] [CrossRef] [Green Version]
  8. Dias, T.; Bronze, M.R.; Houghton, P.J.; Mota-Filipe, H.; Paulo, A. The flavonoid-rich fraction of Coreopsis tinctoria promotes glucose tolerance regain through pancreatic function recovery in streptozotocin-induced glucose-intolerant rats. J. Ethnopharmacol. 2010, 132, 483–490. [Google Scholar] [CrossRef]
  9. Wang, T.; Xi, M.; Guo, Q.; Shen, Z. Chemical components and antioxidant activity of volatile oil of a Compositae tea (Coreopsis tinctoria Nutt.) from Mt. Kunlun. Ind. Crop. Prod. 2015, 67, 318–323. [Google Scholar] [CrossRef]
  10. Zhang, Y.; Shi, S.; Zhao, M.; Chai, X.; Tu, P. Coreosides A–D, C14-polyacetylene glycosides from the capitula of Coreopsis tinctoria and its anti-inflammatory activity against COX-2. Fitoterapia 2013, 87, 93–97. [Google Scholar] [CrossRef] [PubMed]
  11. Shang, Y.F.; Oidovsambuu, S.; Jeon, J.-S.; Nho, C.W.; Um, B.-H. Chalcones from flowers of Coreopsis lanceolata and thier in vitro antioxidative activity. Planta Med. 2013, 79, 295–300. [Google Scholar]
  12. Kim, B.-R.; Paudel, S.B.; Nam, J.-W.; Jin, C.H.; Lee, I.-S.; Han, A.-R. Constituents of Coreopsis lanceolate flower and their dipeptidyl peptidase IV inhibitory effects. Molecules 2020, 25, 4370. [Google Scholar] [CrossRef]
  13. Kimura, Y.; Hiraoka, K.; Kawano, T.; Fujioka, S.; Shimada, A. Nematicidal activities of acetylene compounds from Coreopsis lanceolata L. J. Biosci. 2008, 63, 843–847. [Google Scholar] [CrossRef]
  14. Okada, Y.; Okita, M.; Murai, Y.; Okano, Y.; Nomura, M. Isolation and identification of flavonoids from Coreopsis lanceolata L. pentals. Nat. Prod. Res. 2014, 28, 201–204. [Google Scholar] [CrossRef]
  15. Tanimoto, S.; Miyazawa, M.; Inoue, T.; Okada, Y.; Nomura, M. Chemical constituents of Coreopsis lanceolata L. and their physiological activities. J. Oleo Sci. 2009, 58, 141–146. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  16. Ma, Z.; Zheng, S.; Han, H.; Meng, J.; Yang, X.; Zeng, S.; Zhou, H.; Jiang, H. The bioactive components of Coreopsis tinctoria (Asteraceae) capitula: Antioxidant activity in vitro and profile in rat plasma. J. Funct. Foods 2015, 20, 575–586. [Google Scholar] [CrossRef]
  17. Kim, H.-G.; Oh, H.-J.; Ko, J.-H.; Jung, Y.S.; Oh, S.-M.; Lee, Y.-G.; Kim, D.-O.; Baek, N.-I. Phenolic compounds from the flowers of Coreopsis lanceolata. J. Appl. Biol. Chem. 2019, 62, 323–326. [Google Scholar] [CrossRef]
  18. Kim, B.-R.; Kim, H.M.; Jin, C.H.; Kang, S.-Y.; Kim, J.-B.; Jeon, Y.G.; Park, K.Y.; Lee, I.-S.; Han, A.-R. Composition and antioxidant activities of volatile organic compounds in radiation-bred Coreopsis cultivars. Plants 2020, 9, 717. [Google Scholar] [CrossRef] [PubMed]
  19. Hou, Y.; Li, G.; Wang, J.; Pan, Y.; Jiao, K.; Du, J.; Chen, R.; Wang, B.; Li, N. Okanin, effective constituent of the flower tea Coreopsis tinctoria, attenuates LPS-induced microglial activation through inhibition of the TLR4/NF-κB signaling pathways. Sci. Rep. 2017, 7, 45705. [Google Scholar] [CrossRef] [Green Version]
  20. Korea Seed & Variety Service. Available online: http://www.seed.go.kr/sites/seed_eng/index..do (accessed on 19 July 2021).
  21. Ali, H.; Ghori, Z.; Sheikh, S.; Gul, A.E. Effects of gamma radiation on crop production. In Crop Production and Global Environmental Issues; Hakeem, K., Ed.; Springer: Cham, Switzerland, 2016; pp. 27–78. [Google Scholar]
  22. Burnett, S.E.; Keever, G.J.; Kessler, J.R.; Cilliam, C.H. Foliar application of plant growth retardants to Coreopsis rosea ‘American dream’. J. Environ. Hort. 2000, 18, 39–62. [Google Scholar] [CrossRef]
  23. Park, K.-Y.; Hwang, H.-J.; Chae, W.-B.; Choi, G.-W. Development of a new Coreopsis variety ‘Uridream Pink’ by gamma-ray irradiation. Kor. J. Hort. Sci. Technol. 2014, 32, 906–911. [Google Scholar]
  24. Sorrie, B.A.; LeBlond, R.J.; Weakley, A.S. Identification, distribution, and habitat of Coreopsis section Eublepharis (Asteraceae) and description of a new species. J. Bot. Res. Inst. Tex. 2013, 7, 299–310. [Google Scholar]
  25. Kessler, J.R., Jr.; Keever, G.J. Plant growth retardants affect growth and flowering of Coreopsis verticillata ‘Moonbeam’. J. Environ. Hortic. 2007, 25, 229–233. [Google Scholar] [CrossRef]
  26. Peng, A.; Lin, L.; Zhao, M.; Sun, B. Classification of edible chrysanthemums based on phenolic profiles and mechanisms underlying the protective effects of characteristic phenolics on oxidatively damaged erythrocyte. Food Res. Int. 2019, 123, 64–74. [Google Scholar] [CrossRef] [PubMed]
  27. Li, Y.; Yang, P.; Luo, Y.; Gao, B.; Sun, J.; Lu, W.; Liu, J.; Chen, P.; Zhang, Y.; Yu, L. Chemical compositions of chrysanthemum teas and their anti-inflammatory and antioxidant properties. Food Chem. 2019, 286, 8–16. [Google Scholar] [CrossRef]
  28. Shen, J.; Hu, M.; Tan, W.; Ding, J.; Jiang, B.; Xu, L.; Hamulati, H.; He, C.; Sun, Y.; Xiao, P. Traditional uses, phytochemistry, pharmacology, and toxicology of Coreopsis tinctoria Nutt.: A review. J. Ethnopharmacol. 2021, 269, 113690. [Google Scholar] [CrossRef]
  29. Zălaru, C.; Crişan, C.; Călinescu, I.; Moldovan, Z.; Ţârcomnicu, I.; Litescu, S.; Tatia, R.; Moldovan, L.; Boda, D.; Iovu, M. Polyphenols in Coreopsis tinctoria Nutt. fruits and the plant extracts antioxidant capacity evaluation. Cent. Eur. J. Chem. 2014, 12, 858–867. [Google Scholar] [CrossRef]
  30. UNIFI Scientific Information System, Driver Pack 2020 Release 1; Waters: Milford, MA, USA, 2020.
  31. MassBank. Available online: www.massbank.jp/Search (accessed on 19 July 2021).
  32. Saleem, M.; Hareem, S.; Khan, A.; Naheed, S.; Raza, M.; Hussain, R.; Imran, M.; Choudhary, M.I. Dual inhibitors of urease and carbonic anhydrase-II from Iris species. Pure Appl. Chem. 2019, 91, 1695–1707. [Google Scholar] [CrossRef]
  33. Wollenweber, E.; Valant-Vetschera, K.M.; Fernandes, G.W. Chemodiversity of exudate flavonoids in Baccharis concinna and three further south-american Baccharis species. Nat. Prod. Commun. 2006, 1, 627–632. [Google Scholar] [CrossRef] [Green Version]
  34. Wollenweber, E.; Mann, K.; Doerr, M.; Fritz, H.; Roitman, J.N.; Yatskievych, G. Exudate flavonoids in three Ambrosia species. Nat. Prod. Lett. 1995, 7, 109–116. [Google Scholar] [CrossRef]
  35. Abraham, J.; Thomas, T.D. Isolation, characterization and evaluation of antibacterial activity of a flavanone derivative 8-hydroxyl naringenin from Elephantopus scaber Linn. World J. Pharm. Res. 2015, 4, 2232–2240. [Google Scholar]
  36. Rani, G.; Yadav, L.; Kalidhar, S.B. Phytochemical investigation of Citrus sinensis flavedo variety Blood Red. J. Indian Chem. Soc. 2011, 88, 887–888. [Google Scholar]
  37. Wang, Y.-M.; Zhao, J.-Q.; Yang, J.-L.; Tao, Y.-D.; Mei, L.-J.; Shi, Y.-P. Separation of antioxidant and α-glucosidase inhibitory flavonoids from the aerial parts of Asterothamnus centrali-asiaticus. Nat. Prod. Res. 2017, 31, 1365–1369. [Google Scholar] [CrossRef]
  38. Elshamy, A.I.; Mohamed, T.A.; Marzouk, M.M.; Hussien, T.A.; Umeyama, A.; Hegazy, M.E.F.; Efferth, T. Phytochemical constituents and chemosystematic significance of Pulicaria jaubertii E.Gamal-Eldin (Asteraceae). Phytochem. Lett. 2018, 24, 105–109. [Google Scholar] [CrossRef]
  39. Shimokoriyama, M.; Hattori, S. Anthoclor pigments of Cosmos sulphureus, Coreopsis lanceolata, and C saxicola. J. Am. Chem. Soc. 1953, 75, 1900–1904. [Google Scholar] [CrossRef]
  40. Hoffmann, B.; Hoelzl, J. Chalcone glucosides from Bidens pilosa. Phytochemistry 1988, 28, 247–249. [Google Scholar] [CrossRef]
  41. Sato, T.; Nakayama, T.; Kikuchi, S.; Fukui, Y.; Yonekura-Sakakibara, K.; Ueda, T.; Nishino, T.; Tanaka, Y.; Kusumi, T. Enzymatic formation of aurones in the extracts of yellow snapdragon flowers. Plant Sci. 2001, 160, 229–236. [Google Scholar] [CrossRef]
  42. Kaintz, C.; Molitor, C.; Thill, J.; Kampatsikas, I.; Michael, C.; Halbwirth, H.; Rompel, A. Cloning and functional expression in E. coli of a polyphenol oxidase transcript from Coreopsis grandiflora involved in aurone formation. FEBS Lett. 2014, 588, 3417–3426. [Google Scholar]
  43. Li, J.; Wen, Q.; Feng, Y.; Zhang, J.; Luo, Y.; Tan, T. Characterization of the multiple chemical components of Glechomae Herba using ultra high performance liquid chromatography coupled to quadrupole-time-of-flight tandem mass spectrometry with diagnostic ion filtering strategy. J. Sep. Sci. 2019, 42, 1312–1322. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  44. Ammar, S.; Abidi, J.; Vlad Luca, S.; Boumendjel, M.; Skalicka-Wozniak, K.; Bouaziz, M. Untargeted metabolite profiling and phytochemical analysis based on RP-HPLC-DAD-QTOF-MS and MS/MS for discovering new bioactive compounds in Rumex algeriensis flowers and stems. Phytochem. Anal. 2020, 31, 616–635. [Google Scholar] [CrossRef]
  45. Kandil, F.E.; Grace, M.H. Polyphenols from Cornulaca monacantha. Phytochemistry 2001, 58, 611–613. [Google Scholar] [CrossRef]
  46. Ruiz, E.; Donoso, C.; Gonzalez, F.; Becerra, J.; Marticorena, C.; Silva, M. Phenetic relationships between Juan Fernandez and continental Chilean species of Sophora (Fabaceae) based on flavonoid patterns. Bol. Soc. Chil. Quím. 1999, 44, 351–356. [Google Scholar] [CrossRef]
  47. Elgindi, M.R.; Elgindi, O.D.; Mabry, T.J. Flavonoids of Aptenia cordifolia. Asian J. Chem. 1999, 11, 1525–1527. [Google Scholar]
  48. Olennikov, D.N.; Chirikova, N.K.; Kashchenko, N.I. Spinacetin, a New Caffeoylglycoside, and Other Phenolic Compounds from Gnaphalium uliginosum. Chem. Nat. Compd. 2015, 51, 1085–1090. [Google Scholar] [CrossRef]
  49. Parejo, I.; Bastida, J.; Viladomat, F.; Codina, C. Acylated quercetagetin glycosides with antioxidant activity from Tagetes maxima. Phytochemistry 2005, 66, 2356–2362. [Google Scholar] [CrossRef]
  50. Lai, J.-P.; Lim, Y.H.; Su, J.; Shen, H.-M.; Ong, C.N. Identification and characterization of major flavonoids and caffeoylquinic acids in three Compositae plants by LC/DAD-APCI/MS. J. Chromatogr. B 2007, 848, 215–225. [Google Scholar] [CrossRef]
  51. Hung, T.M.; Cuong, T.D.; Nguyen, H.D.; Zhu, S.; Long, P.Q.; Komatsu, K.; Min, B.S. Flavonoid glycosides from Chromolaena odorata leaves and their in vitro cytotoxic activity. Chem. Pharm. Bull. 2011, 59, 129–131. [Google Scholar] [CrossRef] [Green Version]
  52. Saleem, H.; Zengin, G.; Locatelli, M.; Tartaglia, A.; Ferrone, V.; Htar, T.T.; Naidu, R.; Mahomoodally, M.F.; Ahemad, N. Filago germanica (L.) Huds. bioactive constituents: Secondary metabolites fingerprinting and in vitro biological assays. Ind. Crop. Prod. 2020, 152, 112505. [Google Scholar] [CrossRef]
  53. Nicholls, K.W.; Bohm, B.A. Flavonoids and affinities of Coreopsis bigelovii. Phytochemistry 1979, 186, 1076. [Google Scholar] [CrossRef]
  54. Ishimaru, K.; Sadoshima, S.; Neera, S.; Koyama, K.; Takahashi, K.; Shimomura, K. A polyacetylene gentiobioside from hairy roots of Lobelia inflate. Phytochemistry 1992, 31, 1577–1579. [Google Scholar] [CrossRef]
  55. Zhou, Y.; Wang, Y.; Wang, R.; Guo, F.; Yan, C. Two-dimensional liquid chromatography coupled with mass spectrometry for the analysis of Lobelia chinensis Lour. using an ESI/APCI multimode ion source. J. Sep. Sci. 2008, 31, 2388–2394. [Google Scholar] [CrossRef] [PubMed]
  56. Nauck, M. Incretin therapies: Highlighting common features and differences in the modes of action of glucagon-like peptide-1 receptor agonists and dipeptidyl peptidase-4 inhibitors. Diabetes Obes. Metab. 2016, 18, 203–216. [Google Scholar] [CrossRef] [Green Version]
  57. Mentlein, R. Dipeptidyl-peptidase IV (CD26)-role in the inactivation of regulatory peptides. Regul. Pept. 1999, 85, 9–24. [Google Scholar] [CrossRef]
  58. Langley, A.K.; Suffoletta, T.J.; Jennings, H.R. Dipeptidyl peptidase IV inhibitors and the incretin system in type 2 Diabets Mellitus. Pharmacotherapy 2007, 27, 1163–1180. [Google Scholar] [CrossRef]
  59. Gao, Y.; Zhang, Y.; Zhu, J.; Li, B.; Li, Z.; Zhu, W.; Shi, J.; Jia, Q.; Li, Y. Recent progress in natural products as DPP-4 inhibitors. Future Med. Chem. 2015, 7, 1079–1089. [Google Scholar] [CrossRef]
  60. Kalhotra, P.; Chittepu, V.; Osorio-Revilla, G.; Gallardo-Velázquez, T. Structure-activity relationship and molecular docking of natural product library reveal chrysin as a novel dipeptidyl peptidase-4 (DPP-4) inhibitors: An integrated in silico and In Vitro study. Molecules 2018, 23, 1368. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  61. Parmar, H.S.; Jain, P.; Chauhan, D.S.; Bhinchar, M.K.; Munjal, V.; Yusuf, M.; Choube, K.; Tawani, A.; Tiwari, V.; Manivannan, E.; et al. DPP-IV inhibitory potential of naringin: An in silico, in vitro and in vivo study. Diabetes Res. Clin. Pract. 2012, 97, 105–111. [Google Scholar] [CrossRef] [PubMed]
  62. Kim, B.-R.; Kim, H.Y.; Choi, I.H.; Kim, J.-B.; Jin, C.H.; Han, A.-R. DPP-IV Inhibitory Potentials of Flavonol Glycosides Isolated from the Seeds of Lens culinaris: In Vitro and Molecular Docking Analyses. Molecules 2018, 23, 1998. [Google Scholar] [CrossRef] [Green Version]
  63. Kim, B.-R.; Thapa, P.; Kim, H.M.; Jin, C.H.; Kim, S.H.; Kim, J.-B.; Choi, H.J.; Han, A.-R.; Nam, J.-W. Purification of phenylpropanoids from the scaly bulbs of Lilium longiflorum by CPC and determination of their DPP-IV inhibitory potentials. ACS Omega 2020, 5, 4050–4057. [Google Scholar] [CrossRef] [Green Version]
Plants 10 01661 g001a 550Plants 10 01661 g001b 550
Figure 1. Photos of original and mutant cultivars of Coreopsis rosea and Coreopsis verticillata used in this study.
Figure 1. Photos of original and mutant cultivars of Coreopsis rosea and Coreopsis verticillata used in this study.
Plants 10 01661 g001aPlants 10 01661 g001b
Plants 10 01661 g002a 550Plants 10 01661 g002b 550
Figure 2. Principal component analysis (PCA) (a) score plot and (b) loading plot of metabolome analysis of the 32 Coreopsis cultivars; orthogonal partial least-squares discriminant analysis (OPLS-DA) (c) score plot and (d) S-plot show selected markers for differentiating Coreopsis rosea and Coreopsis verticillata; (e) Variable importance plot (VIP) scores of selected markers; (f) variables averages by group of selected potential marker compounds.
Figure 2. Principal component analysis (PCA) (a) score plot and (b) loading plot of metabolome analysis of the 32 Coreopsis cultivars; orthogonal partial least-squares discriminant analysis (OPLS-DA) (c) score plot and (d) S-plot show selected markers for differentiating Coreopsis rosea and Coreopsis verticillata; (e) Variable importance plot (VIP) scores of selected markers; (f) variables averages by group of selected potential marker compounds.
Plants 10 01661 g002aPlants 10 01661 g002b
Plants 10 01661 g003 550
Figure 3. Hierarchical clustering analysis (HCA) with a heatmap from original and mutant cultivars of Coreopsis species.
Figure 3. Hierarchical clustering analysis (HCA) with a heatmap from original and mutant cultivars of Coreopsis species.
Plants 10 01661 g003
Table
Table 1. The list of the original and mutant cultivars of Coreopsis rosea and Coreopsis verticillata used in this study.
Table 1. The list of the original and mutant cultivars of Coreopsis rosea and Coreopsis verticillata used in this study.
Group
(Plant Name)		No.Cultivar NamesRegistration No.Application No.Breeding Process
I
(C. rosea)		1Heaven’s gate--Original cultivar
2Luckyten 63869-Herbicide-induced artificial mutation
3Redfin4408-γ-Irradiated mutation
4Lemon candy4418-γ-Irradiated mutation
5Shiny pink4420-γ-Irradiated mutation
6Uri-dream 013993-Herbicide-induced artificial mutation
7Luckyten54411-Herbicide-induced artificial mutation
8Luckyten94413-Herbicide-induced artificial mutation
9Uri-dream red6001-γ-Irradiated mutation
10Uri-dream 073998-Herbicide-induced artificial mutation
11Uri-dream 063997-Herbicide-induced artificial mutation
12Pink sherbet4415-γ-Irradiated mutation
II
(C. verticillata)		13Citrine--Original cultivar
14Golden ball No.186421-γ-Irradiated mutation
15Golden ball No.216422-γ-Irradiated mutation
16Golden ball No.265995-γ-Irradiated mutation
17Golden ball No.425997-γ-Irradiated mutation
18Golden ball No.485999-γ-Irradiated mutation
III
(C. rosea)		19Pumpkin Pie--Original cultivar
20Gold ring7523-γ-Irradiated mutation
21Golden ring5994-γ-Irradiated mutation
22Mini ball yellow6453-γ-Irradiated mutation
23Box tree6462-γ-Irradiated mutation
24Orange ball6005-γ-Irradiated mutation
IV
(C. verticillata)		25Route 66--Original cultivar
26Golden sunlight-2018-406γ-Irradiated mutation
27Red sunlight-2018-410γ-Irradiated mutation
28Bright sunlight-2018-408γ-Irradiated mutation
29Yellow sunlight-2018-411γ-Irradiated mutation
30Orange sunlight-2018-399γ-Irradiated mutation
V
(C. verticillata)		31Moonbeam--Original cultivar
32Moonlight sonata-2018-401Selection of phenotypic variant
Table
Table 2. Characterization and tentative identification of metabolites found in original and mutant cultivars of Coreopsis rosea and Coreopsis verticillata using ultra-performance liquid chromatography time-of-flight mass spectrometry (UPLC-QTof MS).
Table 2. Characterization and tentative identification of metabolites found in original and mutant cultivars of Coreopsis rosea and Coreopsis verticillata using ultra-performance liquid chromatography time-of-flight mass spectrometry (UPLC-QTof MS).
Peak No.ESI-MS tR (min)Observed Mass (m/z)Caculated Mass (m/z)Error (ppm)Molecular FormulaKey MSE Fragment Ions (m/z)Identification
14.48465.1030465.1039−0.8C21H22O12303.0503, 285.0397, 151.0034, 125.0239Taxifolin-7-O-glucoside
24.50353.0864353.0878−1.4C16H18O9191.0556, 133.0290Chlorogenic acid
34.96465.1030465.1039−0.8C21H22O12303. 0503, 287.0550, 285.0397, 151.0034, 125.0234Taxifolin-3-O-glucoside
45.21329.0865329.0878−1.3C14H18O9167.0338, 151.0026Vanillic acid-4-glucoside
55.74449.1085449.1089−0.4C21H22O11287.0554, 269.0446, 151.0034, 135.0449Flavanomarein
65.85595.1649595.1668−1.9C27H32O15449.1069, 287.0548, 269.0428, 151.0028, 135.0447Isookanin-7-O-rutinoside
75.93609.1454609.1461−0.7C27H30O16447.0932, 285.0392, 151.0033Luteolin-7-O-sophoroside
85.98433.1135433.1140−0.5C21H22O10271.0605, 253.0499, 135.0449Butin-7-O-glucoside
96.17479.0825479.0831−0.6C21H20O13317.0291, 166.99638-Methoxyeriodictyol-7-O-glucoside
106.23463.1239463.1246−0.7C22H24O13301.0708, 165.0188, 135.0449Coreolanceoline B
116.34463.1251463.1246−0.5C22H24O11301.0708, 165.0188, 135.0449Lanceolin
126.48433.1134433.1140−0.6C21H22O10271.0602, 151.0029, 119.0488Naringenin-7-O-glucoside
136.51611.1612611.1618−0.6C27H32O16449.1080, 287.0551, 269.0393, 135.0447Okanin-3′,4′-O-diglucoside
146.52595.1664595.1668−0.4C27H32O15433.1121, 271.0604, 135.04474′,7,8-Trihydroxyflavone-O-diglucoside
156.58609.1454609.1461−0.7C27H30O16447.0932, 285.0394, 135.0082Fisetin-3,7-O-diglucoside
166.64287.0555287.0561−0.6C15H12O6151.0031, 135.0449Isookanin
176.87303.0502303.0510−0.8C15H12O7285.0399, 151.0084, 135.0447, 125.0240Taxifolin
186.91581.1501581.1512−1.1C26H30O15287.0552, 167.0342, 151.00294′,5,7,8-Tetrahydroxyflavanone-7-O-(6-O-arabinosyl-glucoside)
196.94431.0977431.0984−0.7C21H20O10269.0447, 135.0447, 133.0290Sulfuretin-6-O-glucoside
206.97463.0885463.08820.3C21H20O12301.0346, 151.0031Quercetin-7-O-glucoside
217.01447.0929447.09270.2C21H20O11285.0397, 135.0447, 133.0291Maritimein
227.03447.0929447.09270.2C21H20O11285.0397, 151.0033Luteolin-7-O-glucoside
237.15449.1081449.1089−0.8C21H22O11287.0551, 269.0445, 151.0033, 135.0448Marein
247.26493.0984493.0988−0.4C22H22O13331.0447, 316.0200, 164.9830Taxifolin 3′,7-dimethyl ether 3-O-glucoside
257.33461.1085461.1089−0.4C22H22O11299.0547, 283.0242, 165.0188, 133.02913,3′,4′-Trihydroxy-7-methoxyflavone 3-O-glucoside
267.58641.1141641.1148−0.7C30H26O16317.0294, 301.0342, 285.0381, 179.0343, 161.0224, 135.0447, 133.0289Qurcetagetin-7-O-(6′′-caffeoylglucoside)
277.68515.1183515.1195−1.2C25H24O12353.0869, 191.0557, 179.0346, 135.04473,5-Dicaffeoylquinic acid
287.77269.0449269.0450−0.1C15H10O5135.0447, 133.0287Sulfuretin
297.91431.0978431.0984−0.6C21H20O10285.0398, 151.0031, 133.0289Luteolin-6-O-rhamnoside
308.02433.1135433.1140−0.5C21H22O10271.0606, 135.0448Coreopsin
318.04515.1187515.1195−0.8C25H24O12353.0862, 191.0556, 179.03404,5-Dicaffeoylquinic acid
328.26287.0556287.0561−0.5C15H12O6151.0032, 134.0368, 123.0083Okanin
338.46611.1398611.1406−0.8C30H28O14449.1109, 287.0559, 269.0441, 151.0024Eriodictyol chalcone-O-diglucoside
348.73287.0553287.0561−0.8C15H12O6151.0032, 135.0047Eriodictyol chalcone
358.74299.0555299.0561−0.6C16H12O6284.0319, 151.0032, 133.0291Kaempferide
368.80285.0398285.0405−0.7C15H10O6151.0032, 133.0291Luteolin
378.84477.1396477.1402−0.6C23H26O11315.0864, 300.0624, 282.0527, 148.0524, 135.04354-Methoxylanceoletin-4′-O-glucoside
389.20271.0605271.0612−0.7C15H12O5253.0496, 135.0448,Butein
399.28269.0447269.0450−0.3C15H10O5227.0351, 117.0341Apigenin
409.39831.3595831.3597−0.2C46H56O14785.3536, 666.2998, 545.2401, 145.0291Unknown
419.45557.2244557.2240−2.1C26H38O13233.0650, 191.0554, 149.0441Lobetyolinin
Table
Table 3. Effects of the 70% ethanol extract of Coreopsis cultivars on dipeptidyl peptidase (DPP)-IV activity.
Table 3. Effects of the 70% ethanol extract of Coreopsis cultivars on dipeptidyl peptidase (DPP)-IV activity.
Group
(Plant Name)		No.Cultivar NamesDPP-IV Inhibitory Effects
(IC50, μg/mL) 1
I
(C. rosea)		1Heaven’s gate125.29
2Luckyten 6158.83
3Redfin117.55
4Lemon candy95.39
5Shiny pink76.92
6Uri-dream 0195.53
7Luckyten578.06
8Luckyten978.60
9Uri-dream red66.46
10Uri-dream 07118.13
11Uri-dream 06134.28
12Pink sherbet117.70
II
(C. verticillata)		13Citrine56.86
14Golden ball No.1853.55
15Golden ball No.2149.64
16Golden ball No.2663.84
17Golden ball No.4245.01
18Golden ball No.4841.44
III
(C. rosea)		19Pumpkin Pie76.40
20Gold ring87.62
21Golden ring89.22
22Mini ball yellow74.57
23Box tree76.83
24Orange ball124.88
IV
(C. verticillata)		25Route 6654.87
26Golden sunlight40.37
27Red sunlight45.42
28Bright sunlight47.58
29Yellow sunlight50.45
30Orange sunlight34.01
V
(C. verticillata)		31Moonbeam60.61
32Moonlight sonata61.15
Sitagliptin20.095 (μM)
1 Values are presented as the mean ± SD of three independent experiments. 2 Sitagliptin was used as the positive control.
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Share and Cite

MDPI and ACS Style

Kim, B.-R.; Paudel, S.B.; Han, A.-R.; Park, J.; Kil, Y.-S.; Choi, H.; Jeon, Y.G.; Park, K.Y.; Kang, S.-Y.; Jin, C.H.; et al. Metabolite Profiling and Dipeptidyl Peptidase IV Inhibitory Activity of Coreopsis Cultivars in Different Mutations. Plants 2021, 10, 1661. https://0-doi-org.brum.beds.ac.uk/10.3390/plants10081661

AMA Style

Kim B-R, Paudel SB, Han A-R, Park J, Kil Y-S, Choi H, Jeon YG, Park KY, Kang S-Y, Jin CH, et al. Metabolite Profiling and Dipeptidyl Peptidase IV Inhibitory Activity of Coreopsis Cultivars in Different Mutations. Plants. 2021; 10(8):1661. https://0-doi-org.brum.beds.ac.uk/10.3390/plants10081661

Chicago/Turabian Style

Kim, Bo-Ram, Sunil Babu Paudel, Ah-Reum Han, Jisu Park, Yun-Seo Kil, Hyukjae Choi, Yeo Gyeong Jeon, Kong Young Park, Si-Yong Kang, Chang Hyun Jin, and et al. 2021. "Metabolite Profiling and Dipeptidyl Peptidase IV Inhibitory Activity of Coreopsis Cultivars in Different Mutations" Plants 10, no. 8: 1661. https://0-doi-org.brum.beds.ac.uk/10.3390/plants10081661

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