Application of carrot alkane sesquiterpenoid alcohol or derivative thereof in prevention and treatment of rice bacterial seedling damping-off

Abstract

The invention provides application of carrot alkane sesquiterpenoid dienol or a derivative thereof in prevention and treatment of rice bacterial seedling damping-off, and relates to the technical field of pesticides. Through a series of in-vitro and in-vivo drug effect screening tests, the inventor finds that the carrot alkane sesquiterpenoid dienol has a remarkable inhibition effect on the pathogenicity of the B. plantarii in the in-vitro test and can remarkably inhibit secretion of cycloheptatrienol ketone of the B. plantarii, so that the carrot alkane sesquiterpenoid dienol has a remarkable prevention effect on rice bacterial damping-off caused by the B. plantarii in a rice pot experiment. Therefore, the carrot alkane sesquiterpenoid dienol is an excellent choice developed for preventing and treating rice bacterial seedling damping-off.

Description

Technical Field

[0001] The invention relates to the technical field of pesticides, in particular to application of carotene sesquiterpenoid dienol or its derivatives in preventing and controlling bacterial blight of rice seedlings. Background Art

[0002] Burkholderia plantarii is one of the important pathogens that causes bacterial blight of rice seedlings. It has strong infectivity, fecundity and adaptability, and seriously threatens rice production. In nature, B. plantarii is often distributed in seedbed soil, field water, plant seeds and animal and plant residues. After entering the seedling field, it completes the initial infection under suitable conditions and spreads through seeds and soil to enter the re-infection cycle. The infected seedlings show dwarfing, chlorosis and root growth and development disorders in the early stage, and wither and die in the later stage. Pathological studies have found that B. plantarii secretes a small molecule virulence factor called tropolone, which binds to iron ions to form an insoluble chelate, inducing severe iron deficiency in the roots of plant seedlings, causing the seedlings to grow slowly and eventually wither and die.

[0003] In recent years, the disease has occurred in many major rice-producing areas and has a tendency to spread. Since the rice seedling raising season in China's major rice-producing areas is mostly characterized by high temperature and high humidity, this creates favorable conditions for the growth, reproduction and spread of B. plantarii. In particular, the intensification of the global greenhouse effect and the promotion of greenhouse rice seedling raising technology in recent years have also exacerbated the outbreak of the disease in my country.

[0004] At present, the pesticides for controlling bacterial diseases on the market mainly include inorganic copper, organic copper and thiazole, but there is no effective control agent for this bacterial disease.

[0005] In view of this, the present invention is proposed Summary of the invention

[0006] The first object of the present invention is to provide a use of carotene sesquiterpenoid diene alcohol or its derivatives in preventing and controlling bacterial blight of rice seedlings, so as to solve the above technical problems.

[0007] The second object of the present invention is to provide the use of carotene sesquiterpenoid dienol or its derivatives in inhibiting the production of toxins by plant Burkholderia.

[0008] The third object of the present invention is to provide an agent for preventing and treating bacterial blight of rice seedlings.

[0009] The fourth object of the present invention is to provide a method for preventing and treating bacterial blight of rice seedlings.

[0010] In order to achieve the above objectives, the following technical solutions are adopted:

[0011] In a first aspect, the present invention provides the use of carotene sesquiterpenoid diene alcohol or its derivatives in preventing and controlling bacterial blight of rice seedlings;

[0012] The structural formula of the carotene sesquiterpenoid dienol is:

[0013] As a further technical solution, the rice bacterial seedling blight is caused by Burkholderia phytogenes.

[0014] As a further technical solution, the derivatives of carotene sesquiterpenoid dienol include derivatives of salts, esters, amides and glycosides of carotene sesquiterpenoid dienol.

[0015] In a second aspect, the present invention provides the use of carotene sesquiterpenoid dienol or its derivatives in inhibiting the production of toxins by plant Burkholderia;

[0016] The structural formula of the carotene sesquiterpenoid dienol is:

[0017] As a further technical solution, the toxin includes tropolone.

[0018] As a further technical solution, the derivatives of carotene sesquiterpenoid dienol include derivatives of salts, esters, amides and glycosides of carotene sesquiterpenoid dienol.

[0019] In a third aspect, the present invention provides an agent for preventing and controlling bacterial blight of rice seedlings, comprising carotene sesquiterpenoid diene alcohol or a derivative thereof;

[0020] The structural formula of the carotene sesquiterpenoid dienol is:

[0021] As a further technical solution, the derivatives of carotene sesquiterpenoid dienol include derivatives of salts, esters, amides and glycosides of carotene sesquiterpenoid dienol.

[0022] As a further technical solution, it also includes a carrier;

[0023] The carrier includes at least one of water, glucose, lactose, gum arabic, gelatin, mannitol, starch, magnesium trisilicate, talc, keratin or colloidal silicon dioxide.

[0024] In a fourth aspect, the present invention provides a method for preventing and treating bacterial blight of rice seedlings, comprising:

[0025] a. soaking and treating rice seeds with the above-mentioned agent;

[0026] and / or, b. spraying the above-mentioned agent on rice seedlings.

[0027] Compared with the prior art, the present invention has the following beneficial effects:

[0028] The inventors conducted a series of in vitro and in vivo efficacy screening tests and found that carotene sesquiterpenoids had a significant inhibitory effect on the pathogenicity of B. plantarii in in vitro tests, and could significantly inhibit the secretion of cycloheptatrienolone from B. plantarii, and further showed a significant preventive effect on rice bacterial damping-off caused by B. plantarii in rice pot tests. Therefore, carotene sesquiterpenoids will be an excellent choice for development to prevent and treat rice bacterial damping-off. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] In order to more clearly illustrate the specific implementation methods of the present invention or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0030] Figure 1 It is the structural formula of carotene sesquiterpenoid diene alcohol;

[0031] Figure 2 The effect of carotene sesquiterpenoids on toxin secretion;

[0032] Figure 3 This is the growth medium addition method - rice growth statistics chart;

[0033] Figure 4 This is a statistical chart of seed treatment method-rice growth;

[0034] Figure 5 This is a statistical chart of rice growth under the spraying method. DETAILED DESCRIPTION

[0035] The embodiments of the present invention will be described in detail below in conjunction with the embodiments and examples, but it will be appreciated by those skilled in the art that the following embodiments and examples are only used to illustrate the present invention and should not be considered as limiting the scope of the present invention. Based on the embodiments in the present invention, all other embodiments obtained by those of ordinary skill in the art without making creative work are within the scope of protection of the present invention. If specific conditions are not specified, proceed according to normal conditions or conditions recommended by the manufacturer. If the manufacturer is not specified for the reagents or instruments used, they are all conventional products that can be purchased commercially.

[0036] In a first aspect, the present invention provides the use of carotene sesquiterpenoid diene alcohol or its derivatives in preventing and controlling bacterial blight of rice seedlings;

[0037] The structural formula of the carotene sesquiterpenoid dienol is:

[0038] The inventors have found that carotene sesquiterpenoids have a significant inhibitory effect on the pathogenicity of B. plantarii in an in vitro test, and have a significant preventive effect on rice bacterial damping-off caused by B. plantarii in a rice pot test. Therefore, carotene sesquiterpenoids or their derivatives can be used for the prevention and treatment of rice bacterial damping-off.

[0039] In some optional embodiments, the rice bacterial seedling blight is caused by Burkholderia solani.

[0040] In some optional embodiments, the derivatives of carotene dienol include but are not limited to salts, esters, amides and glycoside derivatives of carotene dienol.

[0041] In a second aspect, the present invention provides the use of carotene sesquiterpenoid dienol or its derivatives in inhibiting the production of toxins by plant Burkholderia;

[0042] The structural formula of the carotene sesquiterpenoid dienol is:

[0043] The inventors have found through research that carotene sesquiterpenoid dienol has the effect of inhibiting the secretion of toxins (such as tropone) by plant Burkholderia, and therefore can be used to inhibit the production of toxins by plant Burkholderia.

[0044] In some optional embodiments, the derivatives of carotene dienol include but are not limited to salts, esters, amides and glycoside derivatives of carotene dienol.

[0045] In a third aspect, the present invention provides an agent for preventing and controlling bacterial blight of rice seedlings, comprising carotene sesquiterpenoid diene alcohol or a derivative thereof;

[0046] The structural formula of the carotene sesquiterpenoid dienol is:

[0047] The agent comprises carotene sesquiterpenoids or its derivatives, and thus has all the technical effects of carotene sesquiterpenoids, and can be used for the prevention and treatment of bacterial seedling blight of rice.

[0048] The medicament of the present invention may be in any suitable form, such as solid, semi-solid, liquid or aerosol form.

[0049] In some optional embodiments, the derivatives of carotene dienol include but are not limited to salts, esters, amides and glycoside derivatives of carotene dienol.

[0050] In some optional embodiments, it further comprises a carrier;

[0051] The carrier includes, but is not limited to, at least one of water, glucose, lactose, gum arabic, gelatin, mannitol, starch, magnesium trisilicate, talc, keratin or colloidal silicon dioxide.

[0052] In some optional embodiments, it also includes, but is not limited to, stabilizers, thickeners, colorants and fragrances.

[0053] In a fourth aspect, the present invention provides a method for preventing and treating bacterial blight of rice seedlings, comprising:

[0054] a. soaking and treating rice seeds with the above-mentioned agent;

[0055] and / or, b. spraying the above-mentioned agent on rice seedlings.

[0056] The method provided by the invention can effectively prevent and control bacterial blight of rice seedlings.

[0057] The present invention is further described below by means of specific examples and comparative examples. However, it should be understood that these examples are only used for more detailed description and should not be construed as limiting the present invention in any form.

[0058] Example 1 Inhibitory effect of carotene sesquiterpenoids on B. plantarii toxin secretion

[0059] 1. Cultivation of B. plantarii

[0060] The Burkholderia plantarii strain was isolated from the rice field in Xiaoshan, Zhejiang Province, and was cultured daily in a constant temperature incubator at 25°C. The culture medium used was potato dextrose broth.

[0061] 2B. plantarii inoculation

[0062] Use an inoculation needle to vertically pick up a single colony of B. plantarii and place it in 50 ml potato glucose broth medium for pre-culture. After culturing for 24 hours, take 10 ml of the bacterial solution, centrifuge at 5000 rpm for 5 minutes, discard the supernatant, add an equal amount of clean water to wash, shake and mix, centrifuge at 5000 rpm for 5 minutes, add an equal amount of clean water again and shake and mix, take 100 μL of the mixed bacterial solution and add it to 60 ml MA liquid culture medium for culture. The formula ratio of MA culture medium is 200X inorganic salt solution (NH4H2PO4 0.1%, KCl 0.02%, MgSO4·7H2O 0.02%), 10 g glucose, and 1 L water. Add carotene sesquiterpenoid dienol (structural formula as shown in the figure) into the MA culture medium respectively. Figure 1 The experimental groups were repeated 3 times, and the culture medium containing B. plantarii and carotene sesquiterpenoids was placed in a constant temperature shaking culture at 25°C.

[0063] Extraction and detection of tropolone secreted by 3B. plantarii

[0064] During the culture stage, 10 ml of bacterial solution was pipetted into a 50 ml centrifuge tube every 6 hours. After centrifugation at 5000 rpm for 5 min, the supernatant was transferred to a new centrifuge tube, the pH was adjusted to 3, and then 10 ml of acetonitrile was added, followed by 1 g of NaCl. After thorough shaking and mixing, the solution was separated into two layers. 2 ml of the supernatant was pipetted, passed through a 0.22 μm organic membrane, and then fixed to the volume in an injection bottle for GC-MS / MS analysis and detection.

[0065] Conclusion: The results are as follows Figure 2 As shown in the results, it was found that with the increase of the concentration of carotene sesquiterpenoid, the level of toxin tropone produced by B. plantarii showed a gradient inhibitory effect. When the concentration of carotene sesquiterpenoid was 20 μM, the toxin production level was reduced by 70%; when the concentration of carotene sesquiterpenoid was 200 μM, the toxin production level was reduced by 75%.

[0066] Example 2 Growth matrix addition method

[0067] 1. Germination of rice seedlings

[0068] Select a sufficient number of rice seeds, place them in a glass culture dish, add a small amount of water (not covering the surface of the seeds), and then culture them in a constant temperature incubator at 25°C. The water needs to be replaced every other day. After 2-3 days, the seeds will turn white, and the germination is complete.

[0069] 2. Treatment of rice seedlings infected with B. plantarii using carotene

[0070] A certain volume of MS plant culture medium was prepared, and 1.5% gellan gum was added thereto to make it a solid culture medium. Each 20 ml volume was dispensed into culture bottles, and B. plantarii was inoculated in the culture medium in the same manner as described in the second part of Example 1. After the inoculation, 20 μM carotene sesquiterpenoid was added to a portion of the culture medium inoculated with B. plantarii as a treatment group, and an experimental group without any treatment was set as a blank control group. After the setting of each experimental group was completed, a number of healthy rice seeds with the same growth potential were selected from the white seeds, and 12 seeds per bottle were planted in each treated culture medium, and the culture bottles were placed in a constant temperature incubator at 25° C., and the rice seedlings were cultured with 12 hours of light and 12 hours of darkness every day, and their growth conditions were continuously observed. On the third day, the lengths of the roots and leaves of the rice seedlings were measured for statistical analysis.

[0071] The results showed that compared with rice seedlings not inoculated with B. plantarii, rice seedlings inoculated with B. plantarii showed obvious symptoms (yellowing and dwarfing), and the growth of the aboveground part and the root system was significantly inhibited; after adding carotene sesquiterpenoids to rice seedlings inoculated with B. plantarii, the development of symptoms was significantly alleviated. In addition, statistical analysis of the length of the roots and leaves of rice seedlings showed that ( Figure 3 In the figure, Control: rice seedlings inoculated with B. plantarii only; Treated: rice seedlings inoculated with B. plantarii and added with carotene sesquiterpenoids; Blank: rice seedlings without any treatment). Carotene sesquiterpenoids can effectively inhibit the bacterial blight of rice seedlings caused by B. plantarii.

[0072] Conclusion: By comparing the length of roots and leaves of rice seedlings in different experimental groups ( Figure 3 ), compared with the growth of rice without any treatment, the growth of rice seedlings inoculated with B. plantarii was significantly inhibited (P﹤0.05). In addition, the growth inhibition of rice seedlings inoculated with B. plantarii and treated with carotene sesquiterpenoids was significantly alleviated (P﹤0.05). This shows that carotene sesquiterpenoids can effectively prevent and control the damping-off disease of rice seedlings caused by B. plantarii infection.

[0073] Example 3 Seed treatment method

[0074] The implementation process was the same as in Example 2, except that the treatment method was changed to using a 20 μM carotene sesquiterpenoid solution to soak the rice seeds. The rest of the experimental setting process and conditions were the same as in Example 2. Figure 4 shown.

[0075] Embodiment 4 spraying method

[0076] The implementation process was the same as in Example 2, except that the treatment method was changed to spraying 20 μM carotene sesquiterpenoid solution on rice seedlings after one day of growth, and the rest of the experimental setting process and conditions were the same as in Example 2. Figure 5 shown.

[0077] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. Application of carotene sesquiterpenoids or their derivatives in preventing and controlling bacterial blight of rice seedlings; The structural formula of the carotene sesquiterpenoid dienol is:

2. The use according to claim 1, characterized in that: The rice bacterial seedling blight is caused by plant Burkholderia.

3. The use according to claim 1, characterized in that: The derivatives of carotene sesquiterpenoid include salts, esters, amides and glycoside derivatives of carotene sesquiterpenoid.

4. Application of carotene sesquiterpenoid diene alcohol or its derivatives in inhibiting the production of toxins by plant Burkholderia; The structural formula of the carotene sesquiterpenoid dienol is:

5. The use according to claim 4, characterized in that: Such toxins include tropolone.

6. The use according to claim 4, characterized in that: The derivatives of carotene sesquiterpenoid include salts, esters, amides and glycoside derivatives of carotene sesquiterpenoid.

7. A medicament for preventing and treating bacterial blight of rice seedlings, characterized in that: Including carotene sesquiterpenoid diene alcohol or its derivatives; The structural formula of the carotene sesquiterpenoid dienol is:

8. The agent according to claim 7, characterized in that The derivatives of carotene sesquiterpenoid include salts, esters, amides and glycoside derivatives of carotene sesquiterpenoid.

9. The drug according to claim 7, characterized in that Also includes carriers; The carrier includes at least one of water, glucose, lactose, gum arabic, gelatin, mannitol, starch, magnesium trisilicate, talc, keratin or colloidal silicon dioxide.

10. A method for preventing and controlling bacterial blight of rice seedlings, characterized in that: include: a. soaking and treating rice seeds with the agent according to any one of claims 7 to 9; And / or, b. spraying rice seedlings with the agent according to any one of claims 7 to 9.

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