Application of spiramerin in improving resistance of rice to rice blast and / or banded sclerotial blight

Abstract

The invention discloses application of spiramerin in improving resistance of rice to rice blast and / or banded sclerotial blight, and relates to the technical field of biology. According to the method, endogenous metabolites capable of influencing the resistance of rice to the rice blast are excavated, the influence of the endogenous metabolites on the disease resistance of the rice is identified in the field through three treatment modes of simultaneous treatment, prevention treatment and treatment, and the result shows that the spiramerin can remarkably enhance the resistance of the rice to the rice blast. The result shows that after spiramerin treatment, pyricularia grisea spore germination and appressorium formation are inhibited. In addition, it is also found that spiramerin has prevention and treatment effects on rice sheath blight disease. The invention provides a novel endogenous metabolite for preventing and treating rice blast and sheath blight, and has important agricultural application value.

Description

Technical Field

[0001] The invention relates to the field of biotechnology, and in particular to the application of spiraea glycoside in improving the resistance of rice to rice blast and / or sheath blight. Background Art

[0002] Rice is one of the main food crops in my country, and rice blast and sheath blight are major diseases of rice, which can cause serious reduction in rice production and sometimes even lead to no harvest. How to prevent and control rice blast and sheath blight has always been the focus and difficulty in the field of rice research, which is of great significance for ensuring my country's food security. At present, breeding disease-resistant rice varieties and using chemical pesticides are common measures to prevent and control rice blast and sheath blight. However, the disease resistance mediated by resistance genes (R genes) usually does not have broad-spectrum resistance and may lose resistance due to changes in the dominant subspecies of pathogens, resulting in large-scale outbreaks of diseases. The long-term use of chemical agents can easily make pathogens resistant, and pesticide residues can affect human and environmental safety. Therefore, it is of great significance to carry out resistance research on rice blast and sheath blight and explore new measures to prevent and control rice blast and sheath blight.

[0003] Biopesticides have attracted extensive attention due to their low toxicity, high efficiency, diverse targets, and environmental protection. Plant secondary metabolites have become one of the important ways to study and create biopesticides because of their non-selectivity to pathogens, low toxicity, easy degradation, and strong targeting. Plant secondary metabolites are usually non-selective to pathogens when improving plant defense capabilities. Compared with chemical agents, biopesticides made from secondary metabolites are not easy to cause pathogens to develop resistance, and there is no safety hazard of pesticide residues. Therefore, screening and identifying key secondary metabolites that enhance disease resistance from metabolites that play an important role in resisting pathogen invasion is of great and far-reaching significance for enhancing crop resistance, stabilizing grain production, and ensuring food security. At present, the secondary metabolites that have been identified to affect the growth and development of pathogens mainly include: Phytocassane AE, Oryzalexin A-E, Sakuranetin, Osthole, Schima superba saponins, and Schizonepeta tenuifolia essential oil. However, compared with the total amount of more than 100,000 secondary metabolites involved in the plant defense system, there is still great potential in exploring key secondary metabolites that can prevent and treat rice blast and sheath blight. The present invention intends to improve rice resistance to rice blast and sheath blight by exploring endogenous metabolites related to rice disease resistance. Summary of the invention

[0004] The purpose of the present invention is to provide an application of spiracoside in improving the resistance of rice to rice blast and / or sheath blight, so as to solve the problems existing in the above-mentioned prior art. The present invention has found that spiracoside can significantly enhance the resistance of rice to rice blast and sheath blight, and has important agricultural application value.

[0005] To achieve the above object, the present invention provides the following solutions:

[0006] The invention provides application of spiracoside in improving resistance of rice to rice blast and / or sheath blight.

[0007] Furthermore, when the spiraea glycoside is used to improve the resistance of rice to rice blast, the spiraea glycoside enhances the resistance of rice to rice blast by inhibiting the spore germination and appressorium formation of the rice blast pathogen.

[0008] The invention also provides the use of spiraea glycoside in preparing pesticide for preventing and treating rice blast and / or sheath blight.

[0009] The invention also provides a pesticide for preventing and controlling rice blast and / or sheath blight, wherein the active ingredient comprises spiraea glycoside.

[0010] Furthermore, the pesticide also includes pesticide-acceptable adjuvants.

[0011] Furthermore, the auxiliary materials include emulsifiers, dispersants and / or co-solvents.

[0012] The present invention also provides the use of the above pesticide in preventing and controlling rice blast and / or sheath blight.

[0013] The present invention also provides a method for preventing and controlling rice blast and / or sheath blight, comprising the step of applying spiraea glycoside to rice plants to prevent and control rice blast and / or sheath blight.

[0014] Furthermore, when controlling rice blast, the application method is foliar spraying; when controlling sheath blight, the application method is stem spraying.

[0015] Furthermore, the spraying concentration of the spiraea glycoside is 200 μM.

[0016] The present invention discloses the following technical effects:

[0017] The present invention explores endogenous metabolites that can affect rice resistance to rice blast, and identifies the effects of endogenous metabolites on rice disease resistance in the field through three treatment methods: simultaneous treatment, preventive treatment, and therapeutic treatment. The results show that spiraea glycoside can significantly enhance rice resistance to rice blast. The results show that after spiraea glycoside treatment, the germination of rice blast spores and the formation of appressorium are inhibited. In addition, the present invention also finds that spiraea glycoside has preventive and therapeutic effects on rice sheath blight. The present invention provides a new endogenous metabolite for preventing and controlling rice blast and sheath blight, and has important agricultural application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0019] Figure 1 Representative pictures of leaves infected with rice blast under simultaneous treatment (A) and statistical diagram of the number of lesions (B);

[0020] Figure 2 Representative pictures of leaves infected with rice blast under the preventive treatment (A) and statistical diagram of the number of lesions (B);

[0021] Figure 3 Representative pictures of leaves infected with rice blast under treatment (A) and statistical diagram of the number of lesions (B);

[0022] Figure 4 The observation diagram of the germination of rice blast fungus spores in different treatment groups (A) and the statistical diagram of the germination rate data (B);

[0023] Figure 5 Representative pictures of stems infected with sheath blight under the preventive treatment (A) and statistical graph of lesion length (B);

[0024] Figure 6 Representative pictures of stems infected with sheath blight under treatment (A) and statistical graphs of lesion lengths (B). DETAILED DESCRIPTION

[0025] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0026] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0027] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0028] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0029] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0030] Spiraeoside referred to in the present invention is a glycosylated flavonoid compound, the English name is Spireoside, abbreviated as Spi, the CAS number is 20229-56-5, and the chemical formula is C 21 H 20 O 12 , MW = 464.38Da, structural formula:

[0031]

[0032] Example 1

[0033] 1. Experimental Materials

[0034] Metabolite: Spi, purchased from Solarbio, product batch number CS30062-69727, was dissolved in dimethyl sulfoxide (DMSO) before use.

[0035] Rice material: Lijiangxintuanheigu (LTH), which is highly susceptible to most physiological races of rice blast fungus. Kitaake, an American rice variety, was used for inoculation with Rhizoctonia solani.

[0036] Pathogen: Pyricularia grisea race Guy11, add spores to sterile water to prepare a spore suspension for later use. Rhizoctonia solani race AG-1-1A, culture on PDA medium containing small wood chips for later use.

[0037] 2. Experimental Methods

[0038] 2.1 Effect of Spi on the incidence of rice blast under different treatments (simultaneous, preventive and therapeutic)

[0039] (1) Simultaneous processing

[0040] LTH rice plants with uniform growth at the four-leaf stage were selected and divided into two groups, and each group was treated as shown in Table 1. After 7 days, based on the number of lesions on the leaves after spraying the pathogen, the incidence of the DMSO-treated group was set as 100%, and the incidence of each treatment group was counted.

[0041] Incidence rate = number of lesions in the experimental group / number of lesions in the DMSO-treated group × 100%.

[0042] Table 1 Experimental groups of simultaneous treatments for rice blast

[0043]

[0044] (2) Preventive treatment

[0045] LTH rice plants with uniform growth at the four-leaf stage were selected and divided into two groups, and each group was treated as shown in Table 2. After 7 days, based on the number of lesions on the leaves after spraying the pathogen, the incidence of the DMSO-treated group was set as 100%, and the incidence of each treatment group was counted.

[0046] Table 2 Experimental groups of rice blast prevention treatments

[0047]

[0048] (3) Treatment

[0049] LTH rice plants with uniform growth at the four-leaf stage were selected and divided into two groups, and each group was treated as shown in Table 3. After 7 days, based on the number of lesions on the leaves after spraying the pathogen, the incidence of the water treatment group was set as 100%, and the incidence of each treatment group was counted.

[0050] Table 3 Experimental groups of rice blast treatments

[0051]

[0052] 2.2 Experiment on inhibition of spore germination of rice blast fungus

[0053] The experiment was divided into three groups: one group was treated with 200μM Spi for Guy11 spore suspension; one group was treated with Guy11 spore suspension without Spi and containing an equal amount of DMSO as solvent control; the other group was treated with Guy11 spore suspension as blank control. The Guy11 spore suspensions of the three groups were dropped on hydrophobic glass slides using the 5-point method, and then placed in an incubator for culture. The germination of spores at 0h, 4h, 12h, and 24h was observed using a Zeiss fluorescence microscope and statistical data were performed.

[0054] 2.3 Effect of Spi on the incidence of rice sheath blight under different treatments (prevention and treatment)

[0055] (1) Preventive treatment

[0056] Kitaake rice plants with uniform growth at the grain filling stage were selected and divided into 2 groups, and each group was treated as shown in Table 4. Five days later, the incidence of the DMSO-treated group was set as 100% based on the length of the lesions on the stem after inoculation, and the incidence of each treatment group was counted.

[0057] Incidence rate = lesion length of experimental group / lesion length of DMSO-treated group × 100%.

[0058] Table 4 Experimental groups of rice sheath blight prevention treatments

[0059]

[0060] (2) Treatment

[0061] Kitaake rice plants with uniform growth at the four-leaf stage were selected and divided into two groups, and each group was treated as shown in Table 5. Five days later, based on the length of the lesions on the stem after inoculation, the incidence of the DMSO-treated group was set as 100%, and the incidence of each treatment group was counted.

[0062] Table 5 Experimental groups of treatments for rice sheath blight

[0063]

[0064] 3. Experimental results

[0065] (1) Statistical results of rice blast incidence

[0066] The statistical results of the incidence of rice blast in each group under different treatments are shown in Table 6, among which the representative pictures of rice blast diseased leaves and the statistical graphs of the number of lesions under simultaneous treatment, preventive treatment and therapeutic treatment are shown in Table 6. Figure 1-Figure 3 The results showed that foliar spraying of Spi could improve the resistance of rice to rice blast, and the incidence of rice blast was significantly reduced.

[0067] Table 6 Statistical results of rice blast incidence under different treatments

[0068] Processing DMSO 200μM Simultaneous processing 100％±25.5％ 51.8％±14.6％ Preventive treatment 100％±26.4％ 52.7％±16.8％ Treatment 100％±33％ 57.5％±17.9％

[0069] (2) Results of spore germination inhibition experiment

[0070] The statistical results of Guy11 spore germination rate in each group under different treatments are shown in Table 7. Among them, the representative pictures and statistical data of the effects of spiraea glycosides on spore germination and appressorium formation of rice blast fungus at different time points under water treatment, DMSO treatment and Spi treatment are shown in Figure 4 The results showed that compared with water treatment and DMSO treatment, the spore germination of Guy11 was significantly inhibited 4h after Spi treatment, and the formation of appressorium was effectively inhibited at 12h and 24h, indicating that Spi can enhance the resistance of rice to rice blast by inhibiting spore germination and appressorium formation, and can be used for the prevention and control of rice blast.

[0071] Table 7 Statistical results of spore germination rate of Guy11 under different treatments

[0072] time Spore state water DMSO Spi 0h Not germinated 99.6％±0.6％ 99.2％±1.2％ 99.2％±1.2％ 4h Germ tube tip swelling 82.1％±2.6％ 79.6％±4.6％ 8.3％±5.2％ 12h Appressorium 85.8％±4.1％ 86.7％±1.2％ 45.4％±0.6％ 24h Mature appressorium 95.8％±1.6％ 92.5％±2.7％ 45.8％±0.6％

[0073] (3) Statistical results of the incidence of sheath blight

[0074] The statistical results of the incidence of sheath blight in each group under different treatments are shown in Table 8. The representative pictures of diseased stems and statistical graphs of lesion length under preventive treatment and therapeutic treatment are shown in Table 8. Figure 5-Figure 6 The results showed that spraying Spi on the stem could improve the resistance of rice to sheath blight.

[0075] Table 8 Statistical results of sheath blight incidence under different treatments

[0076] Processing DMSO 200μM Preventive treatment 100％±19.2％ 60％±17.7％ Treatment 100％±20.2％ 72.8％±24.9％

[0077] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. Application of spiraea glycoside in improving the resistance of rice to rice blast and / or sheath blight.

2. The use according to claim 1, characterized in that: When the spiraea glycoside is used to improve the resistance of rice to rice blast, the spiraea glycoside enhances the resistance of rice to rice blast by inhibiting the spore germination and appressorium formation of the rice blast pathogen.

3. Use of spiraea glycoside in the preparation of pesticides for preventing and controlling rice blast and / or sheath blight.

4. A pesticide for preventing and controlling rice blast and / or sheath blight, characterized in that: Active ingredients include meadowfoam glycosides.

5. The pesticide according to claim 4, characterized in that The pesticide also includes pesticide-acceptable adjuvants.

6. The pesticide according to claim 5, characterized in that The auxiliary materials include emulsifiers, dispersants and / or cosolvents.

7. Use of the pesticide according to any one of claims 4 to 6 in preventing and controlling rice blast and / or sheath blight.

8. A method for preventing and controlling rice blast and / or sheath blight, characterized in that: The method comprises the steps of applying spiraea glycoside to rice plants to prevent and control rice blast and / or sheath blight.

9. The method according to claim 8, characterized in that When controlling rice blast, the application method is foliar spraying; when controlling sheath blight, the application method is stem spraying.

10. The method according to claim 9, characterized in that The spraying concentration of the spiraea glycoside is 200 μM.

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