Functional components of cymbopogon distans volatile oil and application thereof
A technology of rue vanilla and volatile oil, applied in applications, chemicals for biological control, animal repellents, etc., can solve problems such as pollution and affecting the quality of edible fungi
Active Publication Date: 2021-02-09
LANZHOU UNIVERSITY
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AI-Extracted Technical Summary
Problems solved by technology
Most of these miscellaneous bacteria grow rapidly, and if the prevention is not timely, the quality of edible fungi will be seriously affected
Although chemically synthesized drugs to kill plant pathogenic fungi play an irreplaceable role in the prevention and control of plant diseases,...
Abstract
The invention provides application of cymbopogon distans volatile oil in inhibition of phytopathogens or preparation of a botanical bactericide, wherein the phytopathogens are aspergillus flavus, rhizopus oryzae, mucor racemosus, trichoderma viride, lentil acanthomonas, fusarium oxysporum and/or potato late blight bacteria. Activity inhibition experiments prove that the cymbopogon distans volatileoil has high inhibition rate on rhizopus oryzae, mucor racemosus, trichoderma viride and lentil acanthomonas. In addition, the applicant also respectively tests the inhibition activity of main chemical components in the cymbopogon distans plant on the seven fungi, the result shows that the same compound shows different degrees of inhibition effects on different pathogenic bacteria and different inhibition effects on different compounds of the same pathogenic bacteria, and part of the compounds show an obvious inhibition effect, so that the cymbopogon distans volatile oil and the main chemicalcomponents can be used as botanical bactericides with great prospects.
Application Domain
BiocideFungicides +2
Technology Topic
Mucor racemosusBlight +13
Image
Examples
- Experimental program(2)
Example Embodiment
[0041]Example 1
[0042]Test pathogenic fungi: Aspergillus flavus, Rhizopusstolonifer, Mucor racemosus, Trichoderma viride, Colletotrichum lentis, Fusarium oxysporum Fusarium oxysporum and Phytophythora infestans.
[0043]Plant tested: Cymbopogon distans, which is a plant that is sandwiched in a book to make it scent.
[0044]The preparation method of the volatile oil of Rutabaga is: put 250 grams of Rutabaga plant into a round bottom flask, add 350g of distilled water, use steam distillation to collect the volatile oil, the collected oil-water mixture is extracted with n-hexane, and then with anhydrous sulfuric acid Sodium is dried, and finally freeze-dried with a freeze dryer to obtain volatile oil, which is stored in a refrigerator at 4°C.
[0045]experimental method:
[0046]The growth rate method of toxic medium was used to determine the inhibitory effect of the volatile oil of Rutabaga on the growth of plant pathogenic fungi.
[0047]Weigh 8 mg of the volatile oil of Rutabaga, and dissolve it with 100 microliters of DMSO; add it to 20 ml of molten PDA medium to prepare a medicated PDA plate containing a certain concentration of the volatile oil for Rutabaga. Cultured and activated the hyphae blocks of various pathogenic fungi (Aspergillus flavus, Rhizopus staphylococcus, Mucor racemosa, Trichoderma viride, Colletotrichum lunata, Fusarium oxysporum and Phytophthora infestans) with a diameter of 4mm Move to the center of the medicine-containing plate, and set three replicates for each concentration. Place at 18°C (for Phytophthora infestans in potato) and 25°C (except Phytophthora infestans in potato) constant temperature culture. The diameter of the colony was measured by the cross method and corrected to calculate the percentage of inhibition.
[0048]Experimental results:
[0049]Through the activity inhibition experiment, under the volatile oil concentration of 400ppm, the volatile oil of Rutabaga has no effect on Rhizopus stolonifer, Mucor racemosus, Trichoderma viride and Colletotrichum lentis ) These four pathogenic bacteria have excellent inhibitory activity, and the inhibitory rate is greater than 70%. In particular, it shows very significant inhibitory activity against Rhizopus stolonifer, Mucorracemosus and Trichoderma viride, and the inhibitory rates are 96.9%, 95.1%, and 85.0%, respectively. For Aspergillus flavus (Aspergillusflavus), Fusariumoxysporum (Fusariumoxysporum) and Potato infestans (Phytophythora infestans), the inhibition rate is also above 43.1%.
[0050]Table 1 Inhibition rate of volatile oil of Rutabaga volatile oil on 7 kinds of plant pathogens (%, 400ppm)
[0051]
[0052]In order to further confirm whether rutabaga volatile oil has inhibitory effects on Rhizopusstolonifer, Mucor racemosus, Trichoderma viride and Colletotrichum lentis at low concentrations, we The antibacterial activity was further tested at 200ppm, 100ppm and 50ppm.
[0053]The results showed that the inhibitory effect of the volatile oil of the volatile oil on Rhizopusstolonifer was 65.0% at a concentration of 200ppm; the volatile oil of the volatile oil inhibited Rhizopus stolonifer at a concentration of 100ppm. The effect is 51.0%; at a concentration of 50ppm of volatile oil, the volatile oil of Rutabaga has an anti-inhibition effect of Rhizopus stolonifer (-10.0%). The specific results are as followsfigure 1 withfigure 2 Shown.
[0054]figure 1 It is the inhibitory activity of Rhizopus stolonifer volatile oil against Rhizopus stolonifer under different concentration gradients.
[0055]figure 2 It is the inhibition rate of Rhizopus stolonifer in different concentrations of volatile oil of Rhizopus stolonifer.
[0056]At a concentration of 200ppm of Rutabaga volatile oil, Rutabaga volatile oil has the most significant inhibitory effect on Mucorracemosus (84.6%); at a concentration of 100ppm, Rutabaga volatile oil inhibits Mucorracemosus (Mucorracemosus) The effect is 35.2%; at a concentration of 50ppm of volatile oil, the inhibitory effect of Rutabaga volatile oil on Mucor racemosus (Mucor racemosus) is 12.5%; the bacteriostatic effect is positively correlated with the concentration, that is, it gradually decreases as the concentration gradient decreases. Specific results such asimage 3 withFigure 4 Shown.
[0057]image 3 It is the inhibitory activity of Rutabaga volatile oil against Mucor racemosus under different concentration gradients.
[0058]Figure 4 It is the inhibitory rate of different concentrations of Rutabaga volatile oil on Mucor racemosus.
[0059]At a concentration of 200ppm of rutabaga, the inhibitory activity of rutabaga volatile oil on Trichoderma viride is 83.1%; while at lower concentrations (100ppm and 50ppm), rutabaga volatile oil inhibits Trichoderma viride (Trichoderma viride) The activity is very low (6.8% and 4.2% respectively); the specific results are as followsFigure 5 withImage 6 Shown.
[0060]Figure 5 It is the inhibitory activity of the volatile oil of Rutabaga against Trichoderma viride under different concentration gradients.
[0061]Image 6 It is the inhibition rate of different concentrations of Rutabaga volatile oil on Trichoderma viride.
[0062]For Colletotrichum lentis, the inhibitory effect of Colletotrichum lentis on Colletotrichum lentis is 64.0% at a concentration of 200ppm of the essential oil of Brassica oleracea; at a concentration of 100ppm, the volatile oil of Colletotrichum lentis has an inhibitory effect on Colletotrichum lentis. The inhibitory effect of spores is 57.8%; at a volatile oil concentration of 50ppm, the inhibitory effect of Rutabaga volatile oil on Colletotrichum lentilus is 24.0%; the bacteriostatic effect is positively correlated with the concentration, that is, it gradually decreases as the concentration gradient decreases. Specific results such asFigure 7 withFigure 8 Shown.
[0063]Figure 7 It is the inhibitory activity of the volatile oil of rutabaga against Colletotrichumlentis under different concentration gradients.
[0064]Figure 8 It is the inhibition rate of different concentrations of Rutabaga volatile oil on Colletotrichum lentilus.
Example Embodiment
[0065]Example 2
[0066]Test fungi: Aspergillusflavus, Rhizopus stolonife, Mucor racemosus, Trichoderma viride, Colletotrichum lentis, Fusarium oxysporum (Fusarium oxysporum) and Potato infestans (Phytophythora infestans).
[0067]The methods to detect the antibacterial activity of plant volatile oil and main compounds are as follows:
[0068]The colony diameter of the hyphae of the seven plant pathogenic bacteria under each treatment was tested in a cross mode. The smaller the diameter, the stronger the inhibitory activity.
[0069]The preparation process of the tested compound 1-4 is as follows. The obtained volatile oil of Russula officinalis was subjected to silica gel column chromatography according to n-hexane and ethyl acetate (volume ratios were 100:0, 50:1, 20:1, 10:1, 8:1, 4:1, 2:1, 1:1, 0:1) gradient elution was performed, and 9 parts of Fr.1-Fr. were obtained respectively.
[0070]The FR.2 part was eluted by silica gel column chromatography (petroleum ether and dichloromethane were eluted at a volume ratio of 100:1) to obtain pure compound 3 (Δ-Jininene, 1-isopropyl-4,7-dimethyl-1) ,2,3,5,6,8a-hexahydronaphthalene).
[0071]The FR.3 part was prepared by thin layer chromatography (the volume ratio of petroleum ether: ethyl acetate was 15:1) to obtain pure compounds 1 (methyleugenol, methyleugenol) and 4 (elemenin, 1, 2, 3-trimethoxy-5-(2-propenyl)-benzene).
[0072]The FR.4 part was eluted by silica gel column chromatography (the volume ratio of petroleum ether: ethyl acetate was 20:1) to obtain pure compound 2 (butylated hydroxytoluene).
[0073]The above test compounds 1-4 are the main compounds in the volatile oil of Rutabaga. The chemical structural formulas are:
[0074]
[0075]experimental method:
[0076]The growth rate method of toxic medium was used to determine the inhibitory effect of the main chemical components in the volatile oil of Rutabaga on the growth of plant pathogenic fungi.
[0077]Weigh 8 mg of compound 1-4 and dissolve it with 100 microliters of DMSO; add it to 20 ml of molten PDA medium to prepare drug-containing PDA plates with a certain compound concentration. Cultured and activated the hyphae blocks of various pathogenic fungi (Aspergillus flavus, Rhizopus staphylococcus, Mucor racemosa, Trichoderma viride, Colletotrichum lunata, Fusarium oxysporum and Phytophthora infestans) with a diameter of 4mm Move to the center of the medicine-containing plate, and set three replicates for each concentration. Place at 18°C (for Phytophthora infestans in potato) and 25°C (except Phytophthora infestans in potato) constant temperature culture. The diameter of the colony was measured by the cross method and corrected to calculate the percentage of inhibition.
[0078]Experimental results:
[0079]The main compounds 1-4 in the volatile oil of Rutabaga have a strong inhibitory effect. Compound 3 has no obvious inhibitory effect on Aspergillus flavus, Fusarium oxysporum and Phytophthora infestans, but also has significant effects on the remaining four pathogens. The inhibitory effect of compound 2 on Rhizopus glabra and Mucor racemosus were very significant, and the inhibition rates were 94.9% and 90.5%; for Trichoderma viride and Colletotrichum lentilus, the compound 1 and 4 showed obvious inhibitory effects, and their inhibitory rates were between 83.8%-90.3%. The two compounds in the volatile oil of Rutabaga have inhibitory effects on these two pathogens.
[0080]Table 2 Inhibition rate of the main chemical components of volatile oil of Rutabaga on 7 kinds of plant pathogens
[0081]
[0082]In order to further confirm whether compounds 1, 2, 4 and the antibacterial drug carbendazim have inhibitory effects on the above-mentioned strong inhibitory pathogens at low concentrations, we conducted further antibacterial activity tests.
[0083]For Rhizopus stolonifer (Picture 9 withPicture 10 ), the inhibitory activity of compound 2 is significantly better than the positive control carbendazim, especially at a concentration of 200ppm, the inhibition rate of compound 2 on the bacteria is as high as 90%, but the inhibition rate of carbendazim is only above 20%; When the concentration is 100ppm, the inhibition rate of compound 2 is close to 60%; even if the concentration reaches 25ppm, the inhibition rate of compound 2 is more than 30%, so for Rhizopusstolonifer, compound 2 can be used as the first-come antibacterial activity Molecules may be further developed into pesticides.
[0084]Picture 9 Inhibitory activity of compound 2 and carbendazim against Rhizopus stolonifer under different concentration gradients
[0085]Picture 10 It is the inhibitory rate of compound 2 and carbendazim at different concentrations against Rhizopus stolonifer.
[0086]For Mucor racemosus (Picture 11 withPicture 12 ), the inhibitory activity of compound 2 is significantly better than the positive control carbendazim, especially at a concentration of 200ppm, the inhibition rate of compound 2 on the bacteria is as high as 80%, but the inhibition rate of carbendazim is close to 20%; When the concentration is 100ppm, the inhibition rate of compound 2 is also close to 40%; when the concentration reaches 25ppm, the inhibition rate of compound 2 and carbendazim is similar, less than 10%. Therefore, for Mucorracemosus, compound 2 can be used as the first-arrival antibacterial active molecule, and may be further developed into a pesticide.
[0087]Picture 11 It is the inhibitory activity of compound 2 and carbendazim against Mucor racemosus under different concentration gradients.
[0088]Picture 12 It is the inhibition rate of compound 2 and carbendazim to Mucor racemosus at different concentrations.
[0089]For Trichoderma viride (Figure 13 withFigure 14 ), when the concentration is 200 ppm, the inhibitory activity of compounds 1 and 4 is not much different, and its value is close to 80%, but the inhibitory performance of the positive drug carbendazim is slightly stronger. Between 25-100ppm, the strong inhibitory effect of carbendazim did not diminish, and its inhibition rate was greater than 90%, while the inhibitory effects of compounds 1 and 4 were worse than that of carbendazim, so for Trichoderma viride ( For Trichoderma viride), compounds 1 and 4 have the potential to act as first-coming antibacterial active molecules at high concentrations.
[0090]Figure 13 It is the inhibitory activity of compounds 1 and 4 and carbendazim against Trichodermaviride under different concentration gradients.
[0091]Figure 14 It is the inhibition rate of different concentrations of compound 1, 4 and carbendazim on Trichoderma viride.
[0092]For Colletotrichum lentis (Figure 15 withFigure 16), when the concentration is 200ppm, the inhibitory activity of compound 1 is close to that of carbendazim and is greater than 80%, while the inhibitory rate of compound 4 is about 70%. Between 25-100 ppm, the strong inhibitory effect of carbendazim did not diminish, and its inhibitory rate was greater than 90%, and as the concentration of Compounds 1 and 4 decreased, the corresponding inhibitory effect also decreased. Therefore, for Colletotrichum lentis, compounds 1 and 4 have the potential as first-coming antibacterial active molecules at high concentrations.
[0093]Figure 15 It is the inhibitory activity of compound 1 and 4 under different concentration gradients of Colletotrichum lentis.
[0094]Figure 16It is the inhibitory rate of different concentrations of compound 1, 4 and carbendazim on Colletotrichum lentis.
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