Comparative Studies of Fraxinus Species from Korea Using Microscopic Characterization, Phytochemical Analysis, and Anti-Lipase Enzyme Activity
Abstract
:1. Introduction
2. Results
2.1. Anatomical Characteristics of the Leaf
2.2. Anatomical Characteristics of the Petiole
2.3. Anatomical Characteristics of the Midrib
2.4. Anatomical Characteristics of the Stem Bark
2.5. HPLC-DAD Profiles of Five Fraxinus Plants
2.6. Orthogonal Projections to Latent Structures-Discriminant Analysis (OPLS-DA) Multivariate Statistical Analysis
2.7. Anti-lipase Activity
3. Discussion
4. Materials and Methods
4.1. Chemicals and Reagents
4.2. Plant Materials
4.3. Anatomical Analysis
4.4. Sample Preparation
4.5. HPLC-DAD Analysis
4.6. Pancreatic Lipase Inhibition Assay
4.7. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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Parameters | FC | FM | FR | FS | FSV |
---|---|---|---|---|---|
Number of glandular trichome (in 500 × 500 μm2) | 1.7 ± 0.2c | 2.4 ± 0.4b | 2.5 ± 0.4b | 3.6 ± 0.9b | 7.0 ± 0.4a |
Diameter of glandular trichome (μm) | 41.1 ± 1.9a | 42.8 ± 3.7a | 36.3 ± 0.8b | 42.3 ± 2.8a | 32.3 ± 1.5c |
Number of epidermal cells (in 300 × 300 μm2) | 78.4 ± 11.3c | 148.4 ± 12.5a | 98.5 ± 14.2c | 92.3 ± 4.8c | 122.8 ± 8.2b |
Number of stomata (in 300 × 300 μm2) | 11.7 ± 0.4e | 33.9 ± 4.0a | 19.3 ± 1.8c | 14.7 ± 1.3d | 24.6 ± 3.0b |
Stomatal length (μm) | 37.5 ± 2.0a | 22.8 ± 0.6c | 33.4 ± 0.7b | 34.7 ± 2.3ab | 32.5 ± 1.6b |
Stomatal width (μm) | 26.0 ± 2.4a | 17.1 ± 0.5b | 23.9 ± 1.5a | 24.2 ± 2.5a | 25.0 ± 1.5a |
Stomatal index | 13.0 ± 1.1b | 18.7 ± 1.5a | 16.4 ± 1.8a | 13.7 ± 1.3b | 16.7 ± 0.9a |
Parameters | FC | FM | FR | FS | FSV |
---|---|---|---|---|---|
Size of epidermal cell of adaxial portion (length × width, μm) | 17.4 ± 1.3a × 17.8 ± 1a | 11.2 ± 1c × 11.8 ± 1b | 19.4 ± 2.3a × 15.6 ± 1.7ab | 13.8 ± 1.3b × 12.4 ± 2.2b | 12.9 ± 1b × 11.6 ± 0.9b |
Size of epidermal cell of abaxial portion (length × width, μm) | 13.3 ± 1.4b × 13.6 ± 1.6b | 10.2 ± 1.0c × 11.5 ± 1b | 21.1 ± 2.3a × 18.7 ± 3.5a | 17.5 ± 1.9a × 14.9 ± 1.0ab | 13.6 ± 0.9b × 12.4 ± 0.8b |
Ratio of cortex thickness to radius in abaxial portion | 0.23 ± 0.03b | 0.29 ± 0.01a | 0.30 ± 0.04a | 0.28 ± 0.02a | 0.27 ± 0.01a |
Diameter of parenchyma cell of cortex in adaxial portion (μm) | 35.3 ± 2.5a | 24.8 ± 1.9b | 24.9 ± 2.0b | 24.7 ± 1.5b | 20.0 ± 0.4c |
Diameter of parenchyma cell in abaxial portion (μm) | 51.6 ± 4.6a | 42.5 ± 1.8b | 40.4 ± 2.7b | 33.4 ± 2.0c | 27.7 ± 1.2d |
Number of sclerenchyma cell (in 300 × 300 μm2) | 40.9 ± 5.3d | 51.3 ± 3.2c | 70.5 ± 4.7b | 54.9 ± 3.0c | 80.3 ± 3.4a |
Diameter of parenchyma cell in pith (μm) | 81.3 ± 2.6a | 54.3 ± 3.0b | 54.5 ± 10.9b | 37.7 ± 2.7c | 30.9 ± 2.3d |
Parameters | FC | FM | FR | FS | FSV |
---|---|---|---|---|---|
Size of epidermal cell of adaxial portion (length × width, μm) | 21.8 ± 1.5a × 19.1 ± 1.6a | 17.0 ± 2.5b × 14.9 ± 0.3b | 21.8 ± 4.0ab × 16.6 ± 2.1ab | 15.0 ± 1.1b × 13.5 ± 1.6b | 13.1 ± 0.4c × 11.9 ± 0.9b |
Size of epidermal cell of abaxial portion (length × width, μm) | 13.6 ± 1.1ab × 14.3 ± 1.0b | 11.3 ± 1.8b × 13.6 ± 1.4b | 16.3 ± 2.0a × 18.8 ± 2.4a | 13.1 ± 1.0b × 15.1 ± 2.2ab | 12.4 ± 0.6b × 13.2 ± 0.5b |
Number of sclerenchyma cell (in 300 × 300 μm2) | 38.6 ± 3.0c | 52.7 ± 9.2b | 70.1 ± 6.5b | 75.5 ± 17.1b | 114.8 ± 18.7a |
Diameter of parenchyma cell in adaxial portion (μm) | 28.4 ± 1.9a | 23.8 ± 1.4ab | 29.9 ± 5.4a | 20.7 ± 2.3b | 18.1 ± 0.8b |
Diameter of parenchyma cell in abaxial portion (μm) | 53.4 ± 2.5a | 43.8 ± 1.2b | 45.2 ± 2.4b | 35.0 ± 2.7c | 29.2 ± 0.7d |
Diameter of parenchyma cell in pith (μm) | 75.7 ± 3.7a | 61.4 ± 4.4b | 49.2 ± 4.1c | 28.4 ± 4.0d | 22.2 ± 3.5d |
Size of palisade cell (length × width, μm) | 48.6 ± 4.6a × 17.4 ± 4.7a | 44.3 ± 4.5ab × 12.5 ± 0.4a | 44.4 ± 6.4ab × 14.2 ± 0.7a | 46.2 ± 3.0a × 14.4 ± 1.5a | 40.8 ± 1.8b × 12.5 ± 0.5a |
Palisade ratio | 1.7 ± 0.1b | 2.0 ± 0.2a | 1.9 ± 0.2ab | 2.1 ± 0.2a | 2.2 ± 0.2a |
Parameters | FC | FM | FR | FS | FSV |
---|---|---|---|---|---|
Size of cork cell (length × width, μm) | 21.2 ± 3.4ab × 53.6 ± 4.9a | 20.1 ± 4.8ab × 40.0 ± 3.6b | 13.6 ± 1.3b × 33.1 ± 2.3c | 16.2 ± 0.9b × 36.8 ± 1.8bc | 22.1 ± 1.1a × 36.2 ± 1.6bc |
Diameter of parenchyma cell (in cortex length × width, μm) | 28.1 ± 2.0a × 55.3 ± 2.2a | 24.8 ± 1.4ab × 44.5 ± 1.9b | 24.5 ± 2.7ab × 51.6 ± 1.5a | 23.0 ± 1.2b × 42.9 ± 2.5b | 22.8 ± 0.7b × 40.6 ± 1.7b |
Number of phloem fiber (in 500 × 500 μm2) | 83.7 ± 4.6c | 85.4 ± 3.2c | 123.0 ± 16.5ab | 138.4 ± 3.5a | 122.7 ± 4.2b |
Number of phloem ray (in 500 × 500 μm2) | 3.4 ± 0.3b | 4.5 ± 0.5ab | 3.3 ± 0.2b | 3.6 ± 0.5b | 4.8 ± 0.2a |
Size of phloem ray (length × width, μm) | 40.5 ± 0.3a × 21.1 ± 2.0a | 34.8 ± 0.8b × 15.0 ± 1.5b | 34.5 ± 4.3ab × 20.9 ± 1.2a | 32.1 ± 4.5b × 15.5 ± 2.1b | 28.3 ± 0.4b × 14.7 ± 0.4b |
Compounds | Season | FC | FM | FR | FS | FSV |
---|---|---|---|---|---|---|
Esculin (1) | Summer | ND* | 0.31 ± 0.04c | 32.5 ± 2.4b | 31.6 ± 2.5b | 38.7 ± 3.1a |
Autumn | ND | 0.24 ± 0.09c | 24.8 ± 0.8a | 28.8 ± 4.8a | 20.8 ± 2.5b | |
Tyrosol (2) | Summer | ND | 0.56 ± 0.11a | ND | ND | ND |
Autumn | ND | 0.48 ± 0.09a | ND | ND | ND | |
Isofraxidin-7-O-β-D-glucopyranoside (3) | Summer | ND | 2.39 ± 0.41a | 0.23 ± 0.04b | ND | ND |
Autumn | ND | 1.66 ± 0.14a | 0.22 ± 0.01b | ND | ND | |
Fraxin (4) | Summer | ND | 14.8 ± 2.2b | 10.8 ± 1.4c | 58.1 ± 3.9a | 61.7 ± 3.9a |
Autumn | ND | 8.32 ± 1.61c | ND | 37.0 ± 2.4a | 24.8 ± 3.0b | |
Mandshurin (5) | Summer | ND | 20.6 ± 0.7a | ND | ND | ND |
Autumn | ND | 10.2 ± 0.5a | ND | ND | ND | |
Fraxetin (6) | Summer | ND | 0.27 ± 0.03b | 0.08 ± 0.00c | 1.39 ± 0.12a | 1.53 ± 0.15a |
Autumn | ND | 0.26 ± 0.01a | ND | ND | ND | |
Calceolarioside A (7) | Summer | 10.9 ± 1.0b | 4.26 ± 1.50c | 2.23 ± 0.42d | 13.5 ± 1.2a | 10.7 ± 0.4b |
Autumn | 7.44 ± 1.23a | 1.96 ± 0.27c | ND | 7.69 ± 1.42a | 2.72 ± 0.53b | |
Calceolarioside B (8) | Summer | ND | 30.4 ± 2.9a | 10.3 ± 0.7b | 10.6 ± 1.0b | 9.11 ± 1.32b |
Autumn | ND | 9.94 ± 1.45a | 4.04 ± 0.27b | 8.90 ± 1.74a | 7.91 ± 1.13a | |
Pinoresinol-4′-O-β-D-glucopyranoside (9) | Summer | ND | 12.1 ± 0.4a | 0.62 ± 0.17d | 1.74 ± 0.06b | 0.82 ± 0.05c |
Autumn | ND | 9.74 ± 0.56a | 0.61 ± 0.06c | 0.77 ± 0.04b | 0.24 ± 0.00d | |
Fraxinol (10) | Summer | ND | 0.53 ± 0.05a | ND | ND | ND |
Autumn | ND | 0.47 ± 0.02a | ND | ND | ND | |
Oleuropein (11) | Summer | ND | 16.9 ± 0.4a | 10.2 ± 2.0b | 5.34 ± 0.99d | 6.77 ± 0.25c |
Autumn | ND | 6.98 ± 0.32a | 5.10 ± 0.97b | 2.97 ± 0.59c | 6.26 ± 0.95ab | |
Ligstroside (12) | Summer | 27.9 ± 1.8a | 15.5 ± 0.9b | 2.11 ± 0.41c | 2.74 ± 0.47c | 1.97 ± 0.22cd |
Autumn | 23.9 ± 0.8a | 14.7 ± 0.7b | ND | 2.51 ± 0.50c | 1.80 ± 0.31c | |
Total coumarins | Summer | ND | 38.9 ± 3.4c | 43.6 ± 3.8c | 91.1 ± 6.5b | 101.9 ± 7.2a |
Autumn | ND | 21.2 ± 2.3d | 25.5 ± 0.8c | 65.8 ± 7.2a | 45.5 ± 5.5b | |
Total phenylethanoids | Summer | 10.9 ± 1.0d | 35.3 ± 4.5a | 12.5 ± 1.1d | 24.1 ± 2.2b | 19.8 ± 1.7c |
Autumn | 7.44 ± 1.23c | 12.4 ± 1.7b | 4.04 ± 0.27d | 16.6 ± 3.2a | 10.6 ± 1.7bc | |
Total secoiridoids | Summer | 27.9 ± 1.8b | 32.4 ± 1.3a | 12.3 ± 2.4c | 8.08 ± 1.46d | 8.74 ± 0.47d |
Autumn | 23.9 ± 0.8a | 21.7 ± 1.0b | 5.08 ± 0.97d | 5.48 ± 1.09d | 8.06 ± 1.26c | |
Total | Summer | 38.8 ± 2.8c | 118.6 ± 9.6a | 69.1 ± 7.5b | 125.0 ± 10.2a | 131.3 ± 9.4a |
Autumn | 31.3 ± 2.0c | 65.0 ± 5.7b | 34.8 ± 2.1c | 88.6 ± 11.5a | 64.5 ± 8.4b |
Botanical Name | Collection Places (Latitude, Longitude) | Collection Year | Specimen No. | Abbreviations |
---|---|---|---|---|
Fraxinus chiisanensis Nakai | Sancheong (35°27′55.9″N, 127°56′06.7″E) (35°27′55.4″N, 127°56′07.9″E) Jiri Mt. (35°14′58.8″N, 127°42′31.2″E) (35°14′58.0″N, 127°42′29.8″E) Pocheon (37°45′07.1″N, 127°09′58.7″E) | 2016–2018 | PGSC-560-1~7 | FC |
F. mandshurica Rupr. | Sancheong (35°14′00.1″N, 127°47′53.9″E) Jinju (35°10′53.3″N, 128°05′42.8″E) (35°13′59.9″N, 127°47′54.1″E) Pocheon (37°44′57.2″N, 127°09′55.7″E) | 2016–2018 | PGSC-561-1~12 | FM |
F. rhynchophylla Hance | Jinju (35°10′53.1″N, 128°05′42.6″E) Jiri Mt. (35°15′02.1″N, 127°42′30.1″E) Youngwol (37°12′18.4″N, 128°26′05.3″E) Pocheon (37°45′10.2″N, 127°09′49.6″E) | 2016–2018 | PGSC-562-1~20 | FR |
F. sieboldiana Blume | Jiri Mt. (35°14′54.5″N, 127°42′18.3″E) Miryang (35°34′06.6″N, 128°59′34.4″E) (35°34′04.7″N, 128°59′34.9″E) Pocheon (37°45′11.9″N, 127°09′54.9″E) | 2016–2019 | PGSC-563-1~15 | FS |
F. sieboldiana var. angustata Blume | Jiri Mt. (35°14′53.7″N, 127°42′30.0″E) Miryang (35°34′05.2″N, 128°59′34.4″E) (35°34′05.7″N, 128°59′31.8″E) Sancheong (35°17′46.8″N, 127°58′29.6″E) Pocheon (37°45′11.4″N, 127°09′56.2″E) | 2016–2019 | PGSC-564-1~15 | FSV |
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Akter, K.-M.; Park, W.S.; Kim, H.-J.; Khalil, A.A.K.; Ahn, M.-J. Comparative Studies of Fraxinus Species from Korea Using Microscopic Characterization, Phytochemical Analysis, and Anti-Lipase Enzyme Activity. Plants 2020, 9, 534. https://doi.org/10.3390/plants9040534
Akter K-M, Park WS, Kim H-J, Khalil AAK, Ahn M-J. Comparative Studies of Fraxinus Species from Korea Using Microscopic Characterization, Phytochemical Analysis, and Anti-Lipase Enzyme Activity. Plants. 2020; 9(4):534. https://doi.org/10.3390/plants9040534
Chicago/Turabian StyleAkter, Kazi-Marjahan, Woo Sung Park, Hye-Jin Kim, Atif Ali Khan Khalil, and Mi-Jeong Ahn. 2020. "Comparative Studies of Fraxinus Species from Korea Using Microscopic Characterization, Phytochemical Analysis, and Anti-Lipase Enzyme Activity" Plants 9, no. 4: 534. https://doi.org/10.3390/plants9040534