Compound with synergistic effect on shenqinmycin, composition and application thereof
阅读说明：本技术 对申嗪霉素具有增效作用的化合物及其组合物与应用 (Compound with synergistic effect on shenqinmycin, composition and application thereof ) 是由 李冠华 周鑫钰 丁文兵 于 2020-12-30 设计创作，主要内容包括：本发明公开了对申嗪霉素具有增效作用的化合物及其组合物与应用,其中所述化合物选自原小檗碱型生物碱、其衍生物或其药用盐中的一种或多种。本发明的化合物可在低剂量的申嗪霉素用量下,对其对茶树真菌的抑制作用产生显著的增效效果。(The invention discloses a compound with synergistic effect on shenqinmycin, a composition and application thereof, wherein the compound is selected from one or more of protoberberine type alkaloid, derivatives thereof or medicinal salts thereof. The compound of the invention can generate obvious synergistic effect on the inhibition effect of the compound on tea tree fungi under the condition of low dosage of shenqinmycin.)
1. A compound with synergistic effect on shenqinmycin, which is selected from one or more of protoberberine type alkaloid, derivatives thereof or medicinal salts thereof.
2. The compound of claim 1, wherein: the protoberberine type alkaloid is selected from one or more of jateorhizine and palmatine.
3. A composition comprising one or more of a protoberberine type alkaloid, a derivative thereof, or a pharmaceutically acceptable salt thereof, and a shenqinmycin.
4. The composition of claim 3, wherein: the protoberberine type alkaloid is selected from one or more of jateorhizine and palmatine.
5. The composition of claim 3, wherein: in the composition, the mass ratio of the shenqinmycin to the protoberberine type alkaloid is 1: 1-1: 30.
6. The composition of claim 5, wherein: in the composition, the mass ratio of the shenqinmycin to the protoberberine type alkaloid is 1: 15-1: 25.
7. Use of a compound according to any one of claims 1 or 2 to enhance the inhibitory effect of shenqinmycin on tea fungi.
8. Use according to claim 7, characterized in that: the mass of the compound is 1-30 times of that of the shenqinmycin.
9. Use of a composition according to any one of claims 3 to 6 for the control of tea mycoses.
10. Use according to any one of claims 7 to 9, wherein: the tea fungus is selected from one or more of tea anthrax, fusarium and phoma.
Technical Field
The invention relates to the technical field of fungal disease control.
Background
Tea tree fungal diseases are diseases which can obviously harm the growth of tea trees or the quality of tea. Among them, the most common tea plant fungi include Colletotrichum sp, Fusarium sp, Phoma sp, etc., which are easily generated in a large area in a high mountain tea garden and damage tea tree buds, leaves, stems and roots, causing great economic loss.
At present, the control of tea fungus diseases is mainly chemical control, for example, a compound bactericide shenqinmycin which is preferably selected in the tea production at present. The shenqinmycin is a biological source pesticide with high efficiency, low toxicity and good environmental compatibility. However, excessive use of the drug tends to cause resistance and drug resistance of germs to a single agent, and thus falls into a vicious circle in which the amount of the bactericide is continuously increased due to resistance, drug resistance, and the like, thereby exacerbating the problem of environmental safety.
Disclosure of Invention
The invention aims to provide a compound capable of generating obvious synergistic effect on shenqinmycin, and the compound can generate obvious synergistic effect on shenqinmycin with low dosage.
The invention also aims to provide a composition which can generate obvious control effect on tea fungus diseases under the condition of containing low dose of shenqinmycin.
The invention also aims to propose a method for applying the compound or the composition.
The invention firstly provides the following technical scheme:
a compound with synergistic effect on shenqinmycin, which is selected from one or more of protoberberine type alkaloid, derivatives thereof or medicinal salts thereof.
According to some preferred embodiments of the invention: the protoberberine type alkaloid is selected from one or more of jateorhizine and palmatine.
The invention also provides the following technical scheme:
a composition comprising one or more of a protoberberine type alkaloid, a derivative thereof, or a pharmaceutically acceptable salt thereof, and a shenqinmycin.
According to some preferred embodiments of the invention: the protoberberine type alkaloid is selected from one or more of jateorhizine and palmatine.
According to some preferred embodiments of the invention: in the composition, the mass ratio of the shenqinmycin to the protoberberine type alkaloid is 1: 1-1: 30.
According to some preferred embodiments of the invention: in the composition, the mass ratio of the shenqinmycin to the protoberberine type alkaloid is 1: 15-1: 25.
The invention further provides application of the compound in enhancing inhibition of shenqinmycin on tea fungi.
According to some preferred embodiments of the invention: in the application, the mass of the compound is 1-30 times of that of the shenqinmycin.
The invention further provides application of the composition in preventing and treating tea mycoses.
According to some preferred embodiments of the invention: the tea fungus is selected from one or more of tea anthrax, fusarium and phoma.
The invention has the following beneficial effects:
in the compound provided by the invention, palmatine (palmatine) and jatrorrhizine (jatrorrhizine) belong to isoquinoline protoberberine alkaloids, and the compound has broad-spectrum antibacterial activity on plant and animal pathogens, can inhibit plant anthracnose, blight and pepper rust disease, but is unknown to have a remarkable synergistic effect on shenqinmycin.
The embodiment of the invention also shows that when the palmatine or jatrorrhizine is independently applied, the palmatine or jatrorrhizine has no obvious inhibiting effect on tea plant fungi, namely, the palmatine or jatrorrhizine has no obvious antibacterial activity on tea plant pathogenic fungi. But after the compound is combined with shenqinmycin, the compound can generate obvious antibacterial activity to tea tree fungi.
The composition disclosed by the invention is composed of low-dose shenqinmycin and a compound which does not have remarkable antibacterial activity on tea tree fungi, and can effectively reduce the risk of drug resistance of the tea tree fungi.
The composition of the invention has high efficiency, low toxicity and good environmental compatibility.
Detailed Description
The present invention is described in detail with reference to the following examples, but it should be understood that the examples are only for illustrative purposes and are not intended to limit the scope of the present invention. All reasonable variations and combinations that fall within the spirit of the invention are intended to be within the scope of the invention.
Palmatine (palmatine) and jatrorrhizine (jatrorrhizine) required in the following examples were extracted and separated from tinospora cochinchinensis by the following procedures:
extracting the dry powder of the stem of the Tinospora sinensis with ethanol and concentrating;
subjecting the paste crude extract obtained by extraction and concentration to liquid-liquid extraction with petroleum ether, ethyl acetate and n-butanol, and separating the n-butanol layer extract by normal phase silica gel column chromatography; the silica gel column is filled with chloroform, wherein the volume ratio of chloroform to methanol is 98: 2-0: 100; after gradient elution, 9 main fractions are obtained;
subjecting the 4 th main fraction to ODS C18Performing column chromatography separation, eluting with a mixed solution of methanol and water at a volume ratio of 30: 70-80: 20, and combining into 5 sub-components after TLC inspection;
recrystallizing the 2 nd subgroup to obtain palmatine (palmatine) with the following chemical formula:
subjecting the 6 th main fraction to ODS C18Performing column chromatography separation, eluting with a mixed solution of methanol and water at a volume ratio of 30: 70-80: 20, and combining into 5 sub-components after TLC inspection;
recrystallizing the 1 st subgroup to obtain jatrorrhizine (jatrorrhizine) with the following chemical formula:
the shenqinmycin used in the following examples had a purity of 99.9%.
The pathogenic bacteria include tea anthrax, fusarium and phoma, and are provided by tea research institute of agricultural academy of sciences, Hunan province.
The following examples test the bactericidal effect of a compound or composition by the following test procedure:
(1) dissolving the raw material components by using an organic solvent dimethyl sulfoxide (DMSO) to prepare a compound or composition solution of the invention;
(2) adding the compound or composition solution to PDA culture medium composed of 1000ml water, 200g potato, 20g glucose and 25g agar to prepare treatment culture medium, and setting DMSO control group;
(3) pouring the treatment culture medium into a 70mm flat plate to prepare a treatment culture medium flat plate; the pathogenic bacteria to be tested are activated at 28 ℃ in advance, a sterilization puncher with the diameter of 5mm is used for punching holes on the edges of activated colonies with uniform growth, and a fungus cake with the diameter of 5mm is inoculated on a treatment culture medium plate. DMSO medium plates were used as controls;
(4) repeating the inoculation for 3 times, and placing the culture medium plate in an incubator at 28 ℃ for dark culture;
(5) measuring the diameter of the cultured colony by a cross method, and calculating the hypha growth inhibition rate and the inhibition medium concentration EC50Value (mg/L).
EC50The determination of (1): the treating concentrations of shenqinmycin are 120, 60, 30, 15 and 7.5 mug/mL respectively, the treating concentrations of jatrorrhizine and palmatine are 2000, 1500, 1000, 500 and 250 mug/m respectively, the hypha growth inhibition rate is calculated, and the EC of each single-dose liquid medicine is calculated50The value is obtained.
Shenqinmycin and jatrorrhizine (palmatine) were mixed at a ratio of 1: 20. compounding at a ratio of 1:10, 1:5, and 1:2.5, performing gradient dilution based on shenqinmycin concentration to obtain medicinal liquid with carbendazim concentration of 120, 60, 30, 15, and 7.5 μ g/mL, calculating hypha growth inhibition rate, and calculating EC of each compounded medicinal liquid50The value is obtained.
Wherein the hypha growth inhibition rate is as follows:
the hypha growth inhibition rate ═ 100% of (control colony diameter-treated colony diameter)/(control colony diameter-cake diameter).
Example 1
Respectively preparing the solutions of the shenqinmycin, the jatrorrhizine and the palmatine through the test steps, and testing the EC of the solutions on different pathogenic bacteria50The results are as follows:
example 2
Respectively preparing shenqinmycin and jatrorrhizine compound solutions and composition solutions of shenqinmycin and jatrorrhizine according to mass ratios of 1:20, 1:10, 1:5 and 1:2.5 through the test steps, and testing the EC of the shenqinmycin and jatrorrhizine compound solutions on different pathogenic bacteria50Values and synergy, wherein the synergy is calculated as:
synergistic multiple ═ EC of each compounded liquid medicine50Value-shenqinmycin EC50value)/EC of each compounded liquid medicine50The value is obtained.
The test results were as follows:
effect on tea anthrax:
effect on fusarium theophyllum:
action on tea Phoma mould:
example 3
Respectively preparing a compound solution of the shenqinmycin and the palmatine and a composition solution of the shenqinmycin and the palmatine according to the mass ratio of 1:20, 1:10, 1:5 and 1:2.5 through the test steps, and testing the EC of the compound solution on different pathogenic bacteria50Value and synergistic multiple.
The test results were as follows:
effect on tea anthrax:
effect on fusarium theophyllum:
action on tea Phoma mould:
example 4
Respectively preparing a compound solution of the shenqinmycin and the palmatine and a composition solution of the shenqinmycin, the jatrorrhizine and the palmatine according to the mass ratio of 1:15:5, 1:10:10, 1:5:15 and 1:8:2 through the test steps, and testing the EC of the compound solution on different pathogenic bacteria50Value and synergistic multiple.
The test results were as follows:
effect on tea anthrax:
solution composition
Mass ratio of the components
EC50(mg/L)
Multiple of effect
Shenqinmycin
60.08
Shenqinmycin, jatrorrhizine and palmatine
1:15:05
7.41
7.11
Shenqinmycin, jatrorrhizine and palmatine
1:10:10
7.41
7.11
Shenqinmycin, jatrorrhizine and palmatine
1:05:15
7.42
7.10
Shenqinmycin, jatrorrhizine and palmatine
1:08:02
11.03
4.45
Shenqinmycin, jatrorrhizine and palmatine
1:05:05
11.56
4.20
Shenqinmycin, jatrorrhizine and palmatine
1:02:08
10.56
4.69
Effect on fusarium theophyllum:
action on tea Phoma mould:
solution composition
Mass ratio of the components
EC50(mg/L)
Multiple of effect
Shenqinmycin
40.61
Shenqinmycin, jatrorrhizine and palmatine
1:15:05
13.76
1.95
Shenqinmycin, jatrorrhizine and palmatine
1:10:10
9.13
3.45
Shenqinmycin, jatrorrhizine and palmatine
1:05:15
8.83
3.60
Shenqinmycin, jatrorrhizine and palmatine
1:08:02
16.43
1.47
Shenqinmycin, jatrorrhizine and palmatine
1:05:05
10.43
2.89
Shenqinmycin, jatrorrhizine and palmatine
1:02:08
8.54
3.76
From examples 1-3, it can be seen that the sole jatrorrhizine or palmatine has no obvious bacteriostatic action on tea fungi, and the sole shenqinmycin has a certain bacteriostatic action on tea fungi, but when either jatrorrhizine or palmatine is combined with shenqinmycin, the formed composition has a significant inhibitory action on tea fungi, and the bactericidal effect can be improved by more than 5 times under certain preferred implementation ratios. Meanwhile, as can be seen from the above examples, the synergistic effect of palmatine or jatrorrhizine on shenqinmycin can be generated under the condition of low content of shenqinmycin.
The above examples are merely preferred embodiments of the present invention, and the scope of the present invention is not limited to the above examples. All technical schemes belonging to the idea of the invention belong to the protection scope of the invention. It should be noted that modifications and embellishments within the scope of the invention may be made by those skilled in the art without departing from the principle of the invention, and such modifications and embellishments should also be considered as within the scope of the invention.
- 上一篇：一种医用注射器针头装配设备
- 下一篇：一种含氯溴异氰尿酸和三环唑的杀菌组合物