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Nematotoxic coumarins from Angelica pubescens Maxim. f. biserrata Shan et Yuan roots and their physiological effects on Bursaphelenchus xylophilus

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Figure 1

Chemical structures of nematicidal compounds 1-4 isolated from Roots of Angelica pubescens Maxim. f. biserrata Shan et Yuan (Duhuo).
Chemical structures of nematicidal compounds 1-4 isolated from Roots of Angelica pubescens Maxim. f. biserrata Shan et Yuan (Duhuo).

Figure 2

Micrographs of pine wood nematodes treated with compounds 1-4 under SEM. (A) Nematodes in control; (B-E) Nematodes treated with osthole 1, columbianadin 2, bergapten 3, xanthotoxin 4, respectively. Minimum scales in A-E are 5, 100, 200 μm, successively.
Micrographs of pine wood nematodes treated with compounds 1-4 under SEM. (A) Nematodes in control; (B-E) Nematodes treated with osthole 1, columbianadin 2, bergapten 3, xanthotoxin 4, respectively. Minimum scales in A-E are 5, 100, 200 μm, successively.

Figure 3

Observation by TEM on pine wood nematodes treated with compounds 1-4. (A) Nematodes in control; (B-E) Nematodes treated with osthole 1, columbianadin 2, bergapten 3, xanthotoxin 4, respectively. Bars represent 2 μm.
Observation by TEM on pine wood nematodes treated with compounds 1-4. (A) Nematodes in control; (B-E) Nematodes treated with osthole 1, columbianadin 2, bergapten 3, xanthotoxin 4, respectively. Bars represent 2 μm.

Figure 4

The effects of nematicidal compounds on PWN enzymes in vivo. Effects on AchE (A) amylase (B) and cellulase (C) of PWN, respectively.
The effects of nematicidal compounds on PWN enzymes in vivo. Effects on AchE (A) amylase (B) and cellulase (C) of PWN, respectively.

Nematotoxic activities against B. xylophilus of compounds 1-4 isolated from the root extracts of Angelica pubescens Maxim. f. biserrata Shan et Yuan.

Compound (200-1,000 μm) LC 50 (72 hr, μM) 95% CLa (μM) χ 2
Osthole 1 489.17 459.09-527.01 1.25
Columbianadin 2 406.74 387.61-422.06 2.14
Bergapten 3 430.08 401.51-458.02 1.13
Xanthotoxin 4 435.66 410.47-459.20 2.02
Aloperineb 369.76 350.07-388.93 2.85

IC50 values of compounds 1-4 on in vitro enzyme activities of B. xylophilus.

Enzyme Compound IC 50 (μM) 95% CLa (μM) χ 2
Osthole 1 3.24 3.01–3.49 2.35
Columbianadin 2 1.62 1.42–1.85 1.01
AchE Bergapten 3 2.28 2.08–2.55 2.93
Xanthotoxin 4 2.40 2.16–2.71 1.86
Galanthamine Hydrobromideb 0.079 0.071–0.086 3.10
Osthole 1 4.89 4.15–5.51 2.12
Columbianadin 2 1.35 1.12–1.59 1.82
Amylase Bergapten 3 1.72 1.64–1.83 0.40
Xanthotoxin 4 3.25 2.79–3.65 3.51
Acarboseb 1.74 1.63–1.88 1.17
Osthole 1 6.13 5.85–6.42 3.01
Columbianadin 2 4.25 3.94–4.49 1.68
Cellulase Bergapten 3 2.09 1.97–2.23 2.21
Xanthotoxin 4 2.35 2.01–2.65 4.01
Copper Sulfate-Ammonia Complexionb 20.10 19.75–20.49 1.82

Nematotoxic activities of the root extracts of Angelica pubescens Maxim. f. biserrata Shan et Yuan against the Pine Wood nematode B. xylophilus.

Corrected mortalitya (%, Mean ± SD)
Sample (1.0 mg/mL) 24 hr 48 hr 72 hr
Ethanol extract 32.50 ± 1.95 c 57.47 ± 1.56 c 85.83 ± 1.60 b
Ethyl acetate fraction 46.93 ± 2.10 c 80.06 ± 1.01 b 95.25 ± 1.16 ab
Aqueous fraction 16.59 ± 0.45 f 27.26 ± 0.99 g 36.55 ± 1.33 e
Fr.1 3.28 ± 0.71 i 10.74 ± 1.11 i 15.48 ± 1.25 h
Fr.2 4.73 ± 1.13 hi 12.78 ± 0.71 i 17.48 ± 1.50 h
Fr.3 10.74 ± 1.27 g 15.54 ± 1.45 h 23.19 ± 1.05 f
Fr.4 20.53 ± 0.66 e 35.68 ± 1.46 e 40.89 ± 0.40 d
Fr.5 56.97 ± 0.94 a 83.78 ± 1.48 a 96.37 ± 1.24 a
Fr.6 49.89 ± 1.19b 76.41 ± 1.22 c 93.58 ± 1.35 b
Fr.7 21.27 ± 1.35 e 33.14 ± 0.39 f 40.76 ± 0.66 d
Fr.8 10.69 ± 1.41 g 17.36 ± 1.60 h 20.27 ± 1.20 g
Fr.9 6.01 ± 80.76 h 11.52 ± 1.49 i 16.74 ± 1.53 h
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