阅读说明：本技术 荜拔酰胺在防治植物细菌性病害中的应用 (Application of piperlongumine in preventing and treating plant bacterial diseases ) 是由 冯岗 张静 朱发娣 叶火春 闫超 于 2021-10-25 设计创作，主要内容包括：本发明公开了荜拔酰胺在防治植物细菌性病害中的应用,所述所述荜拔酰胺的浓度为50～500mg/L,植物细菌性病害的病原菌为水稻白叶枯病菌、柑橘溃疡病菌、甜瓜细菌性果斑病菌、番茄青枯病菌、草莓细菌性角斑病菌、白菜软腐病菌、马铃薯软腐病菌、芒果细菌性角斑病菌的一种或几种。本发明中荜拔酰胺对植物病原细菌具有显著的抑制作用,为农业病害防治中提供了一种天然源杀菌活性物质；荜拔酰胺可用于防治水稻白叶枯病、芒果细菌性黑斑病和甜瓜细菌性果斑病等多种重要的农业病害；将其作为杀菌剂或抑菌剂,其高效、低毒、安全等特点符合当前新农药创制的要求。(The invention discloses an application of piperlongumine in preventing and treating plant bacterial diseases, wherein the concentration of the piperlongumine is 50-500 mg/L, and pathogenic bacteria of the plant bacterial diseases are one or more of rice bacterial blight, citrus canker pathogenic bacteria, melon bacterial fruit blotch pathogenic bacteria, tomato bacterial wilt pathogenic bacteria, strawberry bacterial angular leaf spot pathogenic bacteria, cabbage soft rot pathogenic bacteria, potato soft rot pathogenic bacteria and mango bacterial angular leaf spot pathogenic bacteria. The piperlongumine has a remarkable inhibiting effect on plant pathogenic bacteria, and provides a natural source bactericidal active substance for preventing and treating agricultural diseases; the piperlongumine can be used for preventing and treating various important agricultural diseases such as rice bacterial blight, mango bacterial black spot, melon bacterial fruit blotch and the like; the compound is used as a bactericide or a bacteriostatic agent, and has the characteristics of high efficiency, low toxicity, safety and the like, which meet the requirements of creating new pesticides at present.)

1. The application of piperlongumine in bactericide or bacteriostatic agent for preventing and treating plant bacterial diseases.

2. The application of piperlongumine in a bactericide or bacteriostat for controlling plant bacterial diseases according to claim 1, wherein the bactericide or bacteriostat is used for controlling plant bacterial diseases, and the active ingredient of the bactericide or bacteriostat is piperlongumine with the concentration of 50-500 mg/L.

3. The application of piperlongumine in a bactericide or bacteriostatic agent for controlling plant bacterial diseases according to claim 1, wherein the bactericide or bacteriostatic agent is in the form of a suspending agent, an aqueous emulsion, a microemulsion, an emulsifiable concentrate, a wettable powder or a water dispersible granule.

4. The application of the piperlongumine in a bactericide or bacteriostat for controlling plant bacterial diseases according to claim 1, wherein pathogenic bacteria of the plant bacterial diseases are one or more of rice bacterial blight, rice bacterial streak germs, citrus canker germs, melon bacterial fruit blotch germs, tomato bacterial wilt germs, strawberry bacterial angular leaf spot germs, cabbage black rot germs, Chinese cabbage soft rot germs, potato soft rot germs, and mango bacterial angular leaf spot germs.

5. A bactericide or bacteriostatic agent for preventing and treating plant bacterial diseases is characterized in that the active component contains piperlongumine.

6. The bactericide or bacteriostatic agent for controlling plant bacterial diseases according to claim 5, wherein the concentration of piperlongumine is 50-500 mg/L.

7. The bactericide or bacteriostatic agent for controlling plant bacterial diseases according to claim 5, wherein the bactericide or bacteriostatic agent is in the form of suspending agent, aqueous emulsion, microemulsion, emulsifiable concentrate, wettable powder or water dispersible granule.

8. The bactericide or bacteriostatic agent for controlling plant bacterial diseases according to claim 5, wherein the pathogenic bacteria of plant bacterial diseases are one or more of rice bacterial blight, citrus canker, melon bacterial fruit blotch, tomato bacterial wilt, strawberry bacterial angular leaf spot, cabbage soft rot, potato soft rot, and mango bacterial angular leaf spot.

Technical Field

The invention belongs to the technical field of herbicides, and relates to application of piperlongumine as a bactericide or a bacteriostatic agent in preventing and treating plant bacterial diseases.

Background

Piperlongumine (Piperlongumine), also called piperlongumine, belongs to amide alkaloids, and has the following chemical structure:

the molecular formula of the piperlongumine is C₁₄H₁₉NO₅The relative molecular mass is 317.3410, and is mainly present in the root of Piper longum of Piperaceae. In recent years, with the continuous and intensive research, piperlongumine is found to have important medicinal values, including the treatment of tumors, spleen diseases, chronic malaria, viral hepatitis, cough, stomachache, asthma, bronchitis, diarrhea, respiratory tract infection, glosspalsy, gonorrhea and the like. Further research also finds that the piperlongumine has the effects of resisting anxiety, vascular hyperplasia, platelet aggregation, fungi, inflammation, tumors and the like, wherein the research on the antitumor activity of the piperlongumine is most extensive.

At present, no report on the agricultural activity of the piperlongumine exists at home and abroad, and the inhibitory effect of the piperlongumine on plant pathogenic bacteria is not reported.

Disclosure of Invention

In view of the defects of the prior art, the invention provides the application of piperlongumine as a bactericide or bacteriostatic agent in preventing and treating plant bacterial diseases.

The purpose of the invention is realized by the following technical scheme:

the application of piperlongumine in a bactericide or a bacteriostatic agent for preventing and treating plant bacterial diseases, wherein: the bactericide or the bacteriostatic agent is used for preventing and treating plant bacterial diseases, the active ingredient of the bactericide or the bacteriostatic agent is piperlongumine, and optionally auxiliary agents are added, wherein: the piperlongumine can be obtained by chemical synthesis or separation from piper longum roots, and the concentration of the piperlongumine is 50-500 mg/L; according to actual needs, the suspending agent, the aqueous emulsion, the microemulsion, the missible oil, the wettable powder or the water dispersible granule can be prepared by adopting the conventional method in the field.

The application of piperlongumine as a bactericide or a bacteriostatic agent in preventing and treating plant bacterial diseases, wherein: the active ingredient of the bactericide or the bacteriostatic agent is piperlongumine, and optionally auxiliary agents are added, wherein: the piperlongumine can be obtained by chemical synthesis or separation from piper longum roots, and the concentration of the piperlongumine is 50-500 mg/mL; according to actual needs, the suspending agent, the aqueous emulsion, the microemulsion, the missible oil, the wettable powder or the water dispersible granule can be prepared by adopting the conventional method in the field; the pathogenic bacteria of the plant bacterial diseases are one or more of Xanthomonas oryzae pv. oryzae, Xanthomonas oryzae pv. oryzae, Xanthomonas anoppodis pv. citri, melon bacterial fruit spot (Acidovorax citrulli), Ralstonia solani, strawberry bacterial angular leaf spot (Xanthomonas fragrans), cabbage black rot (Xanthomonas campestris pv. camptosicum), cabbage soft rot (Erwinia carotovora), potato soft rot (Pectobacter carotovorum subsp. brasiliensis), and mango bacterial angular leaf spot (Xanthomonas campestris pv. senticosus).

The invention also aims to provide a bactericide or bacteriostatic agent for preventing and treating plant bacterial diseases, and the active ingredient of the bactericide or bacteriostatic agent contains piperlongumine. Optionally, the concentration of the piperlongumine is 50-500 mg/L.

Compared with the prior art, the invention has the following advantages:

the piperlongumine has a remarkable inhibiting effect on plant pathogenic bacteria, and provides a natural source bactericidal active substance for preventing and treating agricultural diseases; the piperlongumine can be used for preventing and treating various important agricultural diseases such as rice bacterial blight, mango bacterial black spot, melon bacterial fruit blotch and the like; the compound is used as a bactericide or a bacteriostatic agent, and has the characteristics of high efficiency, low toxicity, safety and the like, which meet the requirements of creating new pesticides at present.

Detailed Description

The technical solutions of the present invention are further described below, but not limited thereto, and modifications or equivalent substitutions may be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.

In the research on agricultural bactericidal active substances of piper longum, an activity tracking method is adopted to find that the piperlongum amide has the control effect on plant pathogenic bacteria. The following experiments are taken as examples to verify the effect of piperlongumine in controlling plant diseases.

Sources of piperlongumine

Piperlongumine, CAS number 20069-09-4, purity greater than 98%, purchased from Gooduet bioscience, Inc.

Secondly, determining the inhibitory activity of piperlongumine on plant pathogenic bacteria

1. Test strains

Rice bacterial blight (Xanthomonas oryzae pv. oryzae), citrus canker (Xanthomonas axonopsis pv. citri), melon bacterial fruit blotch (Acidovorax citrulli), tomato bacterial leaf spot (Ralstonia solanacearum), strawberry bacterial angular leaf spot (Xanthomonas fragariae), cabbage soft rot (Erwinia carotovora), potato soft rot (Pectobacterium carotovorum subsp. brasiliensis) and mango bacterial angular leaf spot (Xanthomonas oryzae comprises pv. mangiensis).

2. Test method

2.1 in vitro Activity assay

In order to measure the agricultural bactericidal activity of the piperlongumine, a 96-well plate method (namely in vitro activity detection) is adopted to evaluate the bacteriostatic activity of the piperlongumine on plant pathogenic bacteria. The 96-well plate method is adopted to determine the inhibitory activity of the piperlongumine to the plant pathogenic bacteria, and the method refers to the stone surpassing method (stone surpassing et al, the inhibitory action and mechanism of protocatechuic aldehyde to sakazakii Cronobacter sakazakii [ J ] modern food technology, 2017,33(07):105-111, 62.). The method comprises the following specific steps:

step S1: firstly, adding the prepared bacterial suspension into a 96-well enzyme label plate, wherein each well is 10 mu L;

step S2: adding 10 μ L of bacterial suspension and 200 μ L of liquid culture medium into the sample control group, adding 200 μ L of liquid culture medium into the blank control group, and setting up a control of piperlongumine compound solution at 200 μ L per well;

step S3: blowing and uniformly mixing the medicament solution (the concentration of the piperlongumine is 500mg/L) and the bacterial suspension, putting samples (rice bacterial blight, citrus canker, tomato bacterial wilt, strawberry bacterial angular leaf spot, melon bacterial fruit spot, cabbage soft rot, potato soft rot and mango bacterial angular leaf spot) into an incubator, performing vibration (500r/min) culture, setting the culture temperature to be 28-30 ℃, the culture time to be 18-24 h, measuring the absorbance of bacterial liquid by using a microplate reader, wherein the calculation formula of the inhibition effect of the compound on the bacteria is as follows:

bacteriostatic rate (%) { (control OD)₆₀₀Blank control OD₆₀₀) - (test group OD 600-Compound solution control OD600) }/(control group OD 600-blank control group OD 600). times.100%.

2.2 data analysis

The results of the bacteriostatic activity of the piperlongumine against 8 plant pathogenic bacteria are shown in table 1, and the results show that the tested piperlongumine with the concentration of 500mg/L has good bacteriostatic action on rice bacterial blight, melon bacterial fruit blotch, citrus canker, tomato bacterial wilt and mango bacterial black spot, the inhibition rate is over 90%, and the piperlongumine has a certain bacteriostatic action on cabbage soft rot bacteria, and the inhibition rate is 77.16%.

TABLE 1 bacteriostatic Activity of Piperlongumine against 8 plant pathogenic bacteria

Test strains	Inhibition ratio (%)	Test strains	Inhibition ratio (%)
				Bacterial blight of rice	90.92	Bacterial angular leaf spot of mango	93.09
Bacterial fruit blotch of melon	93.31	Citrus canker fungus	92.73
				Bacterial wilt of tomato	91.03	Strawberry bacterial angular leaf spot	85.78
Soft rot of potato	84.36	Soft rot of Chinese cabbage	77.16

2.3 potting test method

The in-vivo pot culture test of the piperlongumine on the bacterial fruit blotch of the melon comprises the following specific steps:

step A: uniformly spraying 200mg/L of piperlongumine medicament, 100mg/L of piperlongumine medicament, 50mg/L of piperlongumine medicament, 100mg/L of 4% thiediazole copper and 5 pieces of clean water on the melon (variety: red-son cattle);

and B: spraying and inoculating the suspension of the fruit blotch fungus with the concentration of 1 multiplied by 10 in each pot⁹CFU/mL, each treatment is repeated for 4 times, 5 pots of seedlings are repeated each time, 1 plant is cultivated in each pot, the humidity is kept, the illumination culture is carried out, then the morbidity is observed for 7d and 14d of cultivation, and the disease index and the prevention and treatment effect are calculated [ reference: identification of bacillus BJ _6 and prevention and control of melon bacterial fruit blotch _]The concrete formula is as follows:

disease index ═ Σ (number of diseased leaves at each stage × relative stage value)/(total number of investigated leaves × highest disease value) × 100;

the preventing and treating effect (%) is (contrast disease index-treatment disease index)/contrast disease index x 100.

The leaf diseases are graded as follows [ refer to pesticide effect II, bactericide for preventing and treating cucumber bacterial angular leaf spot bacteria ]:

level 0: no disease spots;

level 1: the lesion area accounts for less than 5% of the whole leaf area;

and 3, level: the lesion area accounts for 6-10% of the whole leaf area;

and 5, stage: the lesion area accounts for 11-20% of the whole leaf area;

and 7, stage: the lesion area accounts for 21-50% of the whole leaf area;

and 9, stage: the lesion area accounts for more than 51% of the whole leaf area.

TABLE 2 protective and therapeutic action of Piper longumine on melon bacterial fruit blotch

The protective and therapeutic effects of piperlongumine on bacterial fruit blotch of melon are shown in table 2. The pot experiment result shows that the piperlongumine has stronger protection effect on the muskmelon bacterial fruit blotch, and the protection effect is 85.86% and 80.29% respectively when the tested concentration is 200mg/L and 100mg/L, which are obviously superior to that of a control medicament, namely thiabendazole copper; the medicine also has stronger treatment effect on the muskmelon bacterial fruit blotch, but the intensity of the medicine is lower than the protection effect, and the treatment effect is 82.90 percent when the test concentration is 200mg/L, which is obviously better than the control medicine of thiediazole copper (73.20 percent). Therefore, the invention provides the application of the piperlongumine in a bactericide or a bacteriostatic agent for preventing and treating plant bacterial diseases, and provides the bactericide or the bacteriostatic agent for preventing and treating plant bacterial diseases, wherein the active ingredient of the bactericide or the bacteriostatic agent contains the piperlongumine. The bactericide or the bacteriostatic agent can be optionally supplemented with an auxiliary agent, piperlongumine can be obtained by chemical synthesis or separation from piper longum roots, and the concentration of the piperlongum is 50-500 mg/L; according to actual needs, the suspending agent, the aqueous emulsion, the microemulsion, the missible oil, the wettable powder or the water dispersible granule can be prepared by adopting the conventional method in the field.

It will be understood by those skilled in the art that the foregoing is only a preferred embodiment of the present invention, and is not intended to limit the invention, and that any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the scope of the present invention.