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. 2019 Dec 24;25(1):65.
doi: 10.3390/molecules25010065.

Inhibitory Effect of Osthole from Cnidium monnieri on Tobacco Mosaic Virus (TMV) Infection in Nicotiana glutinosa

Ya-Han Chen  1   2 , Dong-Sheng Guo  2 , Mei-Huan Lu  2   3 , Jian-Ying Yue  1 , Yan Liu  4 , Chun-Ming Shang  4 , De-Rong An  2 , Ming-Min Zhao  1
Affiliations

Inhibitory Effect of Osthole from Cnidium monnieri on Tobacco Mosaic Virus (TMV) Infection in Nicotiana glutinosa

Ya-Han Chen et al. Molecules. .

Abstract

The coumarin compound of osthole was extracted from Cnidium monnieri and identified by LC-MS and 1H- and 13C-NMR. Osthole was tested for anti-virus activity against tobacco mosaic virus (TMV) using the half-leaf method. The results showed that stronger antiviral activity on TMV infection appeared in Nicotiana glutinosa than that of eugenol and ningnanmycin, with inhibitory, protective, and curative effects of 72.57%, 70.26%, and 61.97%, respectively. Through observation of the TMV particles, we found that osthole could directly affect the viral particles. Correspondingly, the level of coat protein detected by Western blot was significantly reduced when the concentrations of osthole increased in tested plants compared to that of the control. These results suggest that osthole has anti-TMV activity and may be used as a biological reagent to control the plant virus in the half-leaf method.

Keywords: Cnidium monnieri; half-leaf method; inhibitory; osthole; tobacco mosaic virus.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
The HPLC chromatogram of osthole.
Figure 2
Figure 2
Samples and structure of the compound identified from osthole.
Figure 3
Figure 3
The high-performance liquid chromatography/mass spectrometry (HPLC/MS) chromatogram of osthole. (A) The HPLC/MS of chromatogram of osthole. (B) The MS of chromatogram of osthole.
Figure 4
Figure 4
Anti-tobacco mosaic virus (TMV) activities of osthole in N. glutinosa. The half-leaf was smeared with osthole extract mixed with TMV at the same volume, and the right half-leaf was smeared with 40 µL of TMV. (A) Osthole extract (5 mg/mL) and water. (B) Osthole extract (5 mg/mL) and TMV. (C) Ningnanmycin SL (8%; 1000-X dilution) and TMV.
Figure 5
Figure 5
Electron microscopic observation of TMV particles after treatment with osthole for 45 min. The concentration of the purified TMV was 0.60 mg/mL. The sample was observed under 49,000× magnifications using a Hitachi H-600 Electron Microscope. (A) Normal TMV particles. (B) TMV treated with osthole at 3 mg/mL for 45 min. (C) TMV treated with osthole at 5 mg/mL for 45 min.
Figure 6
Figure 6
TMV coat protein (CP) accumulation detected by Western blot analysis.
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