CN111248212A

CN111248212A - Application and method of oleanane pentacyclic triterpene compound in preventing and treating crop bacterial diseases

Info

Publication number: CN111248212A
Application number: CN202010142032.0A
Authority: CN
Inventors: 周学永; 付荣霞; 董建萍
Original assignee: Tianjin Agricultural University
Current assignee: Shaanxi Normal University
Priority date: 2020-03-04
Filing date: 2020-03-04
Publication date: 2020-06-09
Anticipated expiration: 2040-03-04
Also published as: CN111248212B

Abstract

The invention relates to application of an oleanane pentacyclic triterpene compound in preventing and treating crop bacterial diseases and a method for preventing and treating the crop bacterial diseases by using the oleanane pentacyclic triterpene compound. The invention improves the resistance of crops to bacterial diseases by soaking seeds before sowing or soaking roots before transplanting or spraying leaves after transplanting instead of utilizing the direct bacteriostatic action of the oleanane pentacyclic triterpenoid, and finally achieves the aim of resisting diseases. More importantly, the prevention and treatment method provided by the invention can achieve the purpose of reducing bacterial diseases only by applying the pesticide once before planting, thereby greatly reducing the prevention and treatment cost of the conventional pesticide.

Description

Application and method of oleanane pentacyclic triterpene compound in preventing and treating crop bacterial diseases

Technical Field

The invention belongs to the technical field of plant protection, and particularly relates to an application and a method of an oleanane pentacyclic triterpene compound in the aspect of preventing and treating crop bacterial diseases.

Background

Oleanane type also called β -amyrin type, belongs to pentacyclic triterpenoid, which is widely distributed in plant and exists in free or saponin combined state₃β -OH, wherein the A/B, B/C, C/D rings in the 5 six-membered rings are all trans, and the D/E rings are mostly in cis arrangement, the saponin compound containing the pentacyclic triterpene parent nucleus has wide pharmacological action and important biological activity, has the functions of anti-inflammation, liver protection, antivirus, anti-tumor, anti-hypertension, anti-atherosclerosis, anti-arrhythmia, lipid regulation, blood sugar reduction and the like, and is concerned by people.

However, the oleanane pentacyclic triterpenoid is mainly applied to the field of medicines and is less applied to agriculture.

Through searching, no patent publication related to the present patent application has been found.

Disclosure of Invention

The invention aims to overcome the defects in the prior art, and provides the application of the oleanane pentacyclic triterpene compound in the aspect of preventing and treating crop bacterial diseases and a method for preventing and treating crop bacterial diseases by using the same.

The technical scheme adopted by the invention for solving the technical problems is as follows:

application of oleanane pentacyclic triterpene compounds in preventing and treating crop bacterial diseases.

The oleanane pentacyclic triterpene compound comprises three structural types of a compound I, a compound II and a compound III, and the effective component is a mixture of one or more than two structural types, and the basic structural types are as follows:

wherein, R in the structure of the compound III₁、R₂、R₃Represents different radicals, wherein R₁Is OH, R₂Is CH₃Or CH₂OH，R₃Is CH₃Or COOCH₃。

Moreover, the bacterial disease is potato bacterial wilt, tomato bacterial wilt and/or canker or celery soft rot.

Moreover, the applications are:

seed soaking: soaking crop seeds in an oleanane pentacyclic triterpene compound solution for 1-10 minutes; fishing out the soaked seeds, and naturally airing for 1-10 hours to sow;

and/or, transplanting and root soaking: cleaning the roots of the seedlings to be transplanted, soaking the roots of the seedlings in a pentacyclic triterpene compound solution, and maintaining for 1-5 minutes; taking out the seedlings after root soaking, naturally airing for 0.5-4 hours, and transplanting and planting;

and/or, transplanting spray: picking the seedlings to be transplanted from the seedbed, and transplanting according to a conventional method; after at least 1 day of permanent planting, the oleanane-type pentacyclic triterpene compound solution is filled into a sprayer, and the leaves of the seedlings are sprayed with a mist-like solution.

Wherein the preparation steps of the oleanane pentacyclic triterpene compound solution are as follows:

dissolving oleanane type pentacyclic triterpene compound in water to prepare 0.5-10g/L oleanane type pentacyclic triterpene compound solution.

A method for preventing and treating crop bacterial diseases by utilizing oleanane pentacyclic triterpene compounds comprises the following steps:

⑴ preparing solution by dissolving oleanane pentacyclic triterpene compound in water to obtain 0.5-10g/L oleanane pentacyclic triterpene compound solution;

⑵ soaking seeds, soaking seeds in the solution of oleanane pentacyclic triterpene for 1-10 min, taking out the soaked seeds, and naturally airing for 1-10 h to obtain the final product.

⑵ transplanting and soaking the roots, cleaning the roots of the seedlings to be transplanted, soaking the roots of the seedlings in the solution of pentacyclic triterpene compounds for 1-5 minutes, taking out the seedlings after root soaking, naturally airing for 0.5-4 hours, and transplanting and planting.

⑵ transplanting and spraying, collecting the seedling from seedbed, transplanting, planting for at least 1 day, spraying oleanane pentacyclic triterpene compound solution in sprayer, and spraying the seedling leaves with atomized solution.

⑵ soaking seeds, soaking the seeds in the solution of oleanane pentacyclic triterpene for 1-10 min, taking out the soaked seeds, and naturally airing for 1-10 h to sow;

⑶ transplanting and soaking the roots, cleaning the roots of the seedlings to be transplanted, soaking the roots of the seedlings in the solution of pentacyclic triterpene compounds for 1-5 minutes, taking out the seedlings after soaking the roots, naturally airing for 0.5-4 hours, and transplanting and planting;

⑷ transplanting and spraying, collecting the seedling from seedbed, transplanting, planting for at least 1 day, spraying oleanane pentacyclic triterpene compound solution in sprayer, and spraying the seedling leaves with atomized solution.

The invention has the advantages and positive effects that:

1. the invention provides application of oleanane pentacyclic triterpene compounds in preventing and treating crop bacterial diseases, and when the oleanane pentacyclic triterpene compounds are used, the disease resistance of crops can be improved in a seed soaking or root soaking mode before planting, so that the purpose of disease resistance is achieved.

2. The invention provides a method for preventing and treating bacterial diseases by using an oleanane pentacyclic triterpenoid, which is characterized in that the method does not utilize the direct bacteriostatic action of the oleanane pentacyclic triterpenoid, but improves the resistance of crops to the bacterial diseases by soaking seeds before sowing or soaking roots before transplanting or spraying leaves after transplanting, and finally achieves the aim of disease resistance. More importantly, the prevention and treatment method provided by the invention can achieve the purpose of reducing bacterial diseases only by applying the pesticide once before planting, thereby greatly reducing the prevention and treatment cost of the conventional pesticide. The bacterial diseases mainly refer to bacterial diseases such as potato bacterial wilt, tomato bacterial wilt and canker, celery soft rot and the like.

Detailed Description

The following detailed description of the embodiments of the present invention is provided for the purpose of illustration and not limitation, and should not be construed as limiting the scope of the invention.

The raw materials used in the invention are conventional commercial products unless otherwise specified; the methods used in the present invention are conventional in the art unless otherwise specified.

Preferably, the oleanane pentacyclic triterpene compound comprises three structural types of a compound I, a compound II and a compound III, the effective component is a mixture of one or more than two structural types, and the basic structural types are as follows:

Preferably, the bacterial disease is potato bacterial wilt, tomato bacterial wilt and/or canker or celery soft rot.

Preferably, the application is:

More specifically, the preparation and detection are as follows:

example 1

the compound I was weighed and prepared into 1.0 and 2.0g/L solutions with distilled water, respectively. Seed potatoes of the potatoes are cut into 3-4 cm square blocks, are respectively soaked in the compound I solution with two concentrations, are taken out after 1 minute, and are naturally placed on the ground for 2 hours. In the chemical pesticide control test group, potato seed blocks are soaked in 80% mancozeb wettable powder (with the concentration of 0.8g/L) solution, taken out after 1 minute, and naturally aired on the ground for 2 hours. The clear water control group adopts distilled water to soak the potato seed blocks, and the rest treatments are the same.

The potato seed pieces thus treated were sown in a test field. Decomposed organic fertilizer (15000 kg/hm) is applied to test field before sowing²) As base fertilizer, applying 300kg/hm of compound fertilizer during sowing²And (4) performing conventional management. When 4-6 leaves grow out from the potato, the prepared suspension of the Pseudomonas solanacearum is uniformly brushed on the front and the back of the potato leaves by using a sterilizing brush. After that, the disease occurrence of the treated plants was investigated 32 days after inoculation according to the conventional management, and the control effect (%) (disease index in placebo-treated zone) × 100/disease index in placebo) was calculated, and the results are shown in table 1.

TABLE 1 prevention and treatment effects of oleanane-type pentacyclic triterpene Compound I on potato bacterial wilt (seed soaking treatment)

a. ab, c represent statistical analysis results between different treatments, with significance between treatments without the same letter.

As can be seen from the table 1, after the potato leaves are infected by pathogenic bacteria for 32 days, the disease index of the clear water control area is 29.36, the disease index is reduced to 13.44-17.13 after the potato leaves are treated by the solution of the compound I, the disease index is obviously reduced compared with the clear water control area, and the control effect of the potato leaves is 41.66-54.22%. The control effect of 80% mancozeb wettable powder of chemical pesticide is 62.46%, which is higher than that of compound I, but when the concentration of the compound I solution reaches 2.0g/L, the effect is close to that of chemical pesticide.

Example 2

a mixture (1: 1) of the compound I and the compound II is taken and prepared into 0.5 g/L solution, 1.0g/L solution and 2.0g/L solution respectively by using distilled water for later use.

Tomato seeds are sowed in a seedling culture plate, and the seedling culture medium is turf and vermiculite (the ratio is 2: 1). And taking out the tomato seedlings when the tomato seedlings are 3-5 leaves, and transplanting the tomato seedlings into a test field. Applying organic fertilizer and compound fertilizer in the test field, and performing conventional management. After 1 day of transplanting, the tomato seedlings were sprayed on the leaf surface with solutions of compound I and compound II at concentrations of 0.5, 1.0 and 2.0g/L, respectively. The chemical pesticide control group adopts 80% mancozeb wettable powder to prepare a solution with the concentration of 1.3g/L to spray the leaf surfaces of the tomato seedlings. The blank control group was sprayed with clear water.

After 4 days of transplantation, the prepared suspension of Pseudomonas solanacearum (Pseudomonas solanacearum) was uniformly brushed on the front and back of the tomato leaves with a sterilizing brush. After inoculation of the germs, the tomatoes were managed as usual, and after 40 days, when the control tomatoes (which were not treated with compound I and chemical pesticide) had developed sufficiently, the study of the disease of the treated plants was started, and the results are shown in Table 2.

TABLE 2 controlling effect of Compound I + Compound II on tomato bacterial wilt (foliar spray)

a. ab, b represent statistical analysis results between different treatments, with significance between treatments that did not contain the same letter.

As shown in Table 2, after the tomato leaves are infected by pathogenic bacteria for 40 days, the disease index of the clear water contrast is 38.41, and the disease index is reduced to 16.49-19.52 after the tomato leaves are treated by the mixed solution of the compound I and the compound II; after the treatment by using a chemical agent (80 percent mancozeb wettable powder), the disease index is reduced to 13.62. Compared with a clear water control, the oleanane pentacyclic triterpene compound (the mixture of the compound I and the compound II) treated by the method has a more obvious reduction of disease index, but still has a higher morbidity than that treated by chemical pesticides. After the mixture of the compound I and the compound II is treated, the control effect of the tomato bacterial wilt is 49.17-57.07%, and after the mixture is treated by chemical pesticide (80% mancozeb wettable powder is 1.3g/L), the control effect of the tomato bacterial wilt is 64.54%.

The data show that although the control effect of the oleanane pentacyclic triterpene compound (the compound I and the compound II) on the tomato bacterial wilt is lower than that of a chemical pesticide, the control effect still tends to be improved along with the increase of the pesticide concentration, and the oleanane pentacyclic triterpene compound has good application prospect.

Example 3

the compound III is extracted from seeds of traditional Chinese medicine fructus kochiae, and mainly comprises three chemical substances, namely: r₁Is OH, R₂Is CH₃，R₃Is CH₃(ii) a A second substance: r₁Is OH, R₂Is CH₂OH，R₃Is CH₃(ii) a A third substance: r₁Is OH, R₂Is CH₂OH，R₃Is COOCH₃. The compound III is prepared into 1 by using distilled water respectively0g/L of solution.

Tomato seeds are sowed in a seedling culture plate, and the seedling culture medium is turf and vermiculite (the ratio is 2: 1). When the seedling age of the tomato is 3-5 leaves, taking out the tomato seedling, removing soil at the root as much as possible, and treating by adopting the following two modes:

(1) and (5) root soaking treatment. The tomato seedling roots were immersed in a solution of compound III (concentration 1.0g/L) for a period of 1-2 minutes. In the test group adopting chemical pesticide control, the tomato seedling roots are soaked in the mancozeb wettable powder (with the concentration of 1.3g/L) solution for 1-2 minutes. And taking out the tomato seedlings after root soaking, placing the tomato seedlings on the ground, naturally placing the tomato seedlings for 0.5 hour, and then transplanting the tomato seedlings into a test field.

(2) And (5) spraying the leaf surfaces. The tomato seedlings are directly transplanted into the test field. Applying organic fertilizer and compound fertilizer in the test field, and performing conventional management. After 1 day of transplanting, the tomato seedlings were foliar-sprayed with a solution of compound III (concentration 1.0 g/L). The chemical pesticide control group adopts 80% mancozeb wettable powder to prepare a solution with the concentration of 1.3g/L to spray the leaf surfaces of the tomato seedlings. The blank control group was sprayed with clear water.

After 4 days of transplantation, the prepared suspension of tomato canker pathogenic bacteria (Clavibacterium microorganismssp.) was evenly brushed on the front and back of tomato leaves with a sterile brush. After inoculation of the germs, the tomatoes were managed according to the conventional method, and after 60 days, when the control tomatoes (which were not treated with the compound I and the chemical pesticide) were sufficiently attacked, the investigation of the attack of the fruits was started, and the results are shown in Table 3.

TABLE 3 preventive and therapeutic effects of Compound III on tomato canker (root drench and foliar spray)

As can be seen from Table 3, after the tomatoes are infected by the bacterial canker pathogen for 60 days, the fruit disease rate of the clear water control area is 60.3 percent, and after the tomatoes are treated by the compound III solution, the fruit disease rate is reduced to 23.90-27.50 percent; after the treatment by chemical agent (80% mancozeb wettable powder), the fruit disease rate is reduced to 18.90-21.11%. Compared with the clear water control, the fruit disease rate is obviously reduced after the treatment of the compound III solution, but is still higher than that after the treatment of chemical pesticide. After the treatment of the compound III solution, the control effect of the tomato canker is 54.39-60.36 percent, wherein the control effect of the leaf surface spraying is higher than that of the root soaking treatment. After being treated by chemical pesticide (80% mancozeb wettable powder is 1.3g/L), the control effect of the tomato canker is 65.01% -68.66%, and the control effect of leaf surface spraying is higher than that of root soaking application. By analyzing the results, the compound III has better control effect on tomato canker.

Example 4

the compound III is extracted from Kochia scoparia seeds and mainly comprises three chemical substances, namely: r₁Is OH, R₂Is CH₃，R₃Is CH₃(ii) a A second substance: r₁Is OH, R₂Is CH₂OH，R₃Is CH₃；

A third substance: r₁Is OH, R₂Is CH₂OH，R₃Is COOCH₃. Compound III was prepared as a 1.0, 2.0 and 3.0g/L solution with distilled water, respectively.

Before test, 15000kg/hm decomposed organic fertilizer is applied to soil²Applying 300kg/hm of compound fertilizer during sowing². Spreading celery seeds on the seedbed, and covering with a film. When 3-4 true leaves of the celery seedlings are needed, the celery seedlings are picked, soil adhered to the roots of the celery seedlings is carefully removed, and then the celery seedlings are respectively immersed into the compound III solution (the concentration is 1.0g/L, 2.0g/L and 3.0g/L) for 1-2 minutes. In a test group with chemical pesticide control, celery seedlings are soaked in a mancozeb wettable powder (with the concentration of 1.0g/L) solution for 1-2 minutes.

And taking out the celery seedlings after root soaking, placing the celery seedlings on the ground, naturally standing for 0.5 hour, transplanting the celery seedlings into a field, and planting the celery seedlings in three rows in a single ridge, wherein the row spacing is 20cm multiplied by 25 cm. After 3 days of field planting, the suspension of the celery soft rot disease, namely, the Pectobacterium carotovorum spp is uniformly sprayed on the stem and leaf parts of the celery seedlings through a sprayer. After that, according to the conventional management, the occurrence of soft rot of the celery after each treatment is investigated 35 days after the permanent planting, and the results are shown in Table 4.

TABLE 4 field efficacy of Compound III on celery Soft rot

a. b and c represent the statistical analysis results among different treatments, and the treatments without the same letter have significance.

As can be seen from Table 4, the control effect of the compound III on the celery soft rot disease is increased along with the increase of the concentration, when the test concentrations of the compound III are respectively 1.0g/L, 2.0g/L and 3.0g/L, the disease indexes of the celery soft rot disease are respectively 20.37, 18.06 and 15.26, and the disease index of the clear water control is 36.85. When the test concentration of the compound III is increased from 1.0g/L to 3.0g/L, the control effect of the celery leaf spot is increased from 44.72% to 58.59%, wherein when the test concentration reaches 2.0g/L and 3.0g/L, the control effect is obvious, and is gradually close to the control effect of the mancozeb wettable powder (61.03%); when the concentration of the compound III is 3g/L, the control effect of the compound III is not obviously different from that of the control medicament which is treated by 80 percent mancozeb wettable powder (1.0 g/L).

Although the embodiments of the present invention have been disclosed for illustrative purposes, those skilled in the art will appreciate that: various substitutions, changes and modifications are possible without departing from the spirit and scope of the invention and the appended claims, and therefore the scope of the invention is not limited to the embodiments disclosed.

Claims

1. Application of oleanane pentacyclic triterpene compounds in preventing and treating crop bacterial diseases.

2. Use according to claim 1, characterized in that: the oleanane pentacyclic triterpene compound comprises three structural types of a compound I, a compound II and a compound III, the effective component is a mixture of one or more than two structural types, and the basic structural types are as follows:

3. Use according to claim 1, characterized in that: the bacterial diseases are potato bacterial wilt, tomato bacterial wilt and/or canker or celery soft rot.

4. Use according to any one of claims 1 to 3, characterized in that: the application is as follows:

5. A method for preventing and treating crop bacterial diseases by utilizing oleanane pentacyclic triterpene compounds is characterized by comprising the following steps: the method comprises the following steps:

6. A method for preventing and treating crop bacterial diseases by utilizing oleanane pentacyclic triterpene compounds is characterized by comprising the following steps: the method comprises the following steps:

7. A method for preventing and treating crop bacterial diseases by utilizing oleanane pentacyclic triterpene compounds is characterized by comprising the following steps: the method comprises the following steps:

8. A method for preventing and treating crop bacterial diseases by utilizing oleanane pentacyclic triterpene compounds is characterized by comprising the following steps: the method comprises the following steps: