Application of paenibacillus kribbensis spaceflight mutant deltaPS04-17 strain in prevention and treatment of cucumber mosaic virus

Abstract

The invention discloses an application of a paenibacillus kribbensis space mutant PS04-17 strain in prevention and treatment of cucumber mosaic virus. Researches show that the PS04-17 strain has the effect of inhibiting CMV and can significantly reduce the morbidity of CMV host plants; after PS04-17 treatment, the relative control effect on plant diseases caused by CMV reaches 54.72%, the morbidity of the tobacco cucumber mosaic virus disease is reduced, the propagation of CMV in a plant body can be effectively inhibited, the morbidity is delayed, the occurrence of the cucumber mosaic virus disease is prevented, and the premature senility of the plant is prevented; meanwhile, the PS04-17 strain can promote tobacco growth, promote root development, increase the tobacco leaf area, enhance the tobacco growth vigor and increase the yield; the PS04-17 strain is green and safe, not only can reduce the use of pesticides, but also is beneficial to the development of environment safety, can be used for green prevention and control of CMV, and has important significance on prevention and control of tobacco diseases and yield increase.

Description

Technical Field

[0001] The invention belongs to the technical field of plant disease prevention and control, and more specifically, relates to the application of a space mutant ΔPS04-17 strain of Paenibacillus kriebensis in preventing and controlling cucumber mosaic virus. Background Art

[0002] Cucumber mosaic virus (CMV) is a very serious viral disease with the largest host range and the widest distribution. CMV can infect all parts of the plant, from the seedling stage to the adult stage, and is transmitted by aphids and friction. More than 60 aphid species can transmit the virus, mainly tobacco aphids and cotton aphids. CMV can infect about 124 species of plants in 36 families of dicots and 4 families of monocots. CMV can cause a decrease in the yield and quality of its host plants, seriously affecting agricultural production. CMV is the main viral disease on tobacco. The virus is distributed and harmful in all tobacco-growing areas in the world, and can cause tobacco cucumber mosaic virus disease. CMV proliferates and transfers quickly in tobacco plants. After infection, it will appear in the mesophyll cells within 6 hours, which will harm the growth of tobacco, reduce the carbohydrate content of diseased leaves, increase the nitrogen content, and reduce the quality. In my country, the Huanghuai tobacco area is the most seriously affected, followed by the central and southern China tobacco areas. In epidemic years, the yield of severely affected areas is reduced by 25-30%, and in severe cases, the yield is often reduced by about 50%. The disease caused by CMV puts the yield and quality of tobacco cultivation in my country at risk. Therefore, taking effective prevention and control measures is crucial to reduce the harm of cucumber mosaic virus.

[0003] At present, the methods for preventing and controlling CMV include: selecting disease-resistant varieties, strengthening field management, and using pesticides. In addition, cucumber mosaic virus can also be comprehensively prevented and controlled through seed treatment, crop rotation, grafting and disinfection. However, it takes a long time to select disease-resistant varieties, and strengthening field management is time-consuming and laborious. The use of chemical agents, such as plant disease-resistant, Ningnanmycin, and copper hydrochloride, can improve the prevention and control effect by spraying, but it will increase the frequency of pesticide use, increase the drug resistance of the virus, and have adverse effects on the safety of plant planting and the environment. Therefore, there is still a lack of products for preventing and controlling CMV to inhibit CMV and reduce the impact of CMV on plants, reduce the occurrence of its diseases, and improve the yield and quality of its host plants. Summary of the invention

[0004] The technical problem to be solved by the present invention is to overcome the shortcomings of existing CMV prevention and treatment products and provide application of aerospace mutant △PS04-17 strain of Paenibacillus cribenbergii in preventing and treating cucumber mosaic virus.

[0005] The invention aims to provide a space-induced mutant of Paenibacillus kribbensis, ΔPS04-17 strain, for use in preventing and controlling cucumber mosaic virus.

[0006] Another object of the present invention is to provide a method for preventing and controlling cucumber mosaic virus or reducing the incidence of cucumber mosaic virus disease.

[0007] The above-mentioned purpose of the present invention is achieved through the following technical solutions:

[0008] The present invention provides a new application of a space-induced mutant of Paenibacillus kribbensis, △PS04-17 strain, in preventing and controlling cucumber mosaic virus. Studies show that the △PS04-17 strain can significantly reduce the incidence of CMV host plants and has the effect of inhibiting CMV. After the host plant is treated with the fermentation liquid of △PS04-17, the relative prevention effect on the host plant disease tobacco cucumber mosaic virus disease caused by CMV reaches 54.72%, which can well prevent the occurrence of tobacco cucumber mosaic virus disease, prevent premature aging of host plants, effectively inhibit the spread of CMV in the plant body, and delay the onset of the disease; at the same time, the microbial agent prepared by the △PS04-17 strain can effectively promote the growth of tobacco plants, promote the development of tobacco roots, increase the leaf area of ​​tobacco, enhance the growth of tobacco, and increase the yield of tobacco. The space-induced mutant strain △PS04-17 of Paenibacillus kribensis used in the present invention is green and safe, can not only reduce the use of pesticides, but also is conducive to the development of environmental safety, can be used for the green prevention and control of CMV, and is of great significance to the prevention and control of tobacco diseases and the increase of production and efficiency.

[0009] Therefore, the present invention provides the use of the space mutant △PS04-17 strain of Paenibacillus cribenbergii in preventing and controlling cucumber mosaic virus.

[0010] The invention provides application of a space mutant △PS04-17 strain of Paenibacillus cribenbergii in preventing and treating plant diseases caused by cucumber mosaic virus.

[0011] In particular, the present invention shows that the space mutant △PS04-17 strain of Paenibacillus krieben has a certain inhibitory effect on CMV, and also has a good control effect on host plant diseases caused by CMV infection, and can significantly reduce the field incidence of its host plants. Therefore, it should have the same or similar control effect on other plant diseases caused by CMV, so those skilled in the art can expect that the space mutant △PS04-17 strain of Paenibacillus krieben can be used for all plant diseases caused by CMV infection.

[0012] The invention provides application of a space mutant △PS04-17 strain of Paenibacillus cribenbergii in preparing a cucumber mosaic virus prevention and control product.

[0013] The invention provides application of a space mutant ΔPS04-17 strain of Paenibacillus cribenbergii in promoting tobacco growth.

[0014] The present invention provides application of a space mutant ΔPS04-17 strain of Paenibacillus cribenbergii in preparing a growth-promoting agent.

[0015] The invention provides application of a space mutant △PS04-17 strain of Paenibacillus cribenbergii in improving tobacco yield.

[0016] The invention provides a method for preventing and controlling cucumber mosaic virus or reducing the incidence of cucumber mosaic virus disease. The plant is treated by adopting a space mutant △PS04-17 strain of Paenibacillus kriebensis or a bacterial liquid thereof.

[0017] Preferably, the bacterial liquid is a fermentation liquid.

[0018] More preferably, the concentration of the fermentation broth is not less than 1×10 9 CFU / mL.

[0019] Preferably, the fermentation liquid is used to treat the plants by foliar spraying.

[0020] More preferably, the fermentation broth needs to be diluted at least 50 times before use.

[0021] The present invention also provides a method for preparing the fermentation liquid of the △PS04-17 strain: inoculating the △PS04-17 bacterial liquid into the fermentation medium at an inoculation amount of 1-10%, and fermenting at 25-30°C and 150-200rpm for 5-9 days.

[0022] Preferably, the fermentation medium uses a modified Czapek liquid medium: 1.5 g / L sodium nitrate, 1 g / L dipotassium hydrogen phosphate, 0.5 g / L magnesium sulfate, 0.5 g / L potassium chloride, 0.01 g / L ferrous sulfate, and 30 g / L sucrose.

[0023] The present invention has the following beneficial effects:

[0024] The invention provides a new application of a space-induced mutant strain of Paenibacillus cribensis △PS04-17 in preventing and controlling cucumber mosaic virus. Studies have shown that the △PS04-17 strain can significantly reduce the incidence of CMV host plants and has the effect of inhibiting CMV. After the host plant is treated with the fermentation liquid of △PS04-17, the relative prevention effect of tobacco cucumber mosaic virus disease caused by CMV reaches 54.72%, which can prevent the occurrence of tobacco cucumber mosaic virus disease, prevent premature aging of tobacco plants, effectively inhibit the spread of CMV in the plant body, and delay the onset of the disease; at the same time, the microbial agent prepared by the △PS04-17 strain can effectively promote the growth of tobacco plants, promote the development of tobacco roots, increase the area of ​​tobacco leaves, enhance the growth of tobacco, and have obvious effects of increasing production and quality, which can not only reduce the use of pesticides, but also be conducive to the development of environmental safety, and can be used for green prevention and control of CMV, which is of great significance to tobacco disease prevention and control and increasing production and efficiency. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 This is a comparison of tobacco plants in the control group and the treatment group 30 days after inoculation with CMV.

[0026] Figure 2 This is a comparison chart of tobacco plants between the control group and the treatment group.

[0027] Figure 3 This is a comparison of the fresh weight of the aboveground part of tobacco between the control group and the treatment group.

[0028] Figure 4 This is a comparison of the fresh weight of the underground part of tobacco between the control group and the treatment group.

[0029] Figure 5 This is a comparison of the maximum leaf area of ​​tobacco in the control group and the treatment group. DETAILED DESCRIPTION

[0030] The present invention is further described below in conjunction with the accompanying drawings and specific examples, but the examples do not limit the present invention in any form. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art.

[0031] Unless otherwise specified, the reagents and materials used in the following examples are commercially available.

[0032] Example 1 Preparation of microbial agent

[0033] The space-induced mutant strain △PS04-17 of Paenibacillus kribbensis used in this embodiment is the research result of the research team of the applicant's research group. It was deposited in the Guangdong Provincial Microbiological Culture Collection Center on March 17, 2022, with the deposit number: GDMCC NO: 62233, and is recorded in the prior art: CN 114480222 A.

[0034] Preparation of microbial inoculant: △PS04-17 bacterial liquid was inoculated into a modified Czapek liquid medium (1.5 g / L sodium nitrate, 1 g / L potassium dihydrogen phosphate, 0.5 g / L magnesium sulfate, 0.5 g / L potassium chloride, 0.01 g / L ferrous sulfate, 30 g / L sucrose) at an inoculum size of 1%, and fermented at 25-30°C and 150 rpm for 7 days to prepare a fermentation liquid containing the space-induced mutant strain of Paenibacillus kriebenensis △PS04-17. The fermentation liquid concentration was 1×10 9 CFU / mL.

[0035] Example 2 Control of Cucumber Mosaic Virus

[0036] Since the virus cannot be cultured in vitro, it needs a host to work. Cucumber mosaic virus (CMV) has a wide host range and can cause a variety of plant diseases. It can cause tobacco cucumber mosaic virus disease on the host plant tobacco. The microbial agent containing the space-induced mutant strain △PS04-17 of Paenibacillus cribensis prepared in Example 1 was used to conduct a pot test on its host plant. The pot test was conducted at South China Agricultural University in Guangdong Province. The test plant in this example was tobacco, and the tobacco seeds were provided by the Plant Pathology Laboratory of South China Agricultural University.

[0037] After sowing tobacco seeds, place them in a constant temperature culture room for cultivation, water them regularly, and transplant them after germination. When the plants grow to 4 to 5 leaves, select plants with consistent growth conditions and no diseases or insect pests and divide them into two groups: a control group and a treatment group.

[0038] To prepare CMV virus stock solution, first cut fresh leaves with CMV virus, then mix them with PBS buffer (0.01mol / L, pH 7.2) in a certain ratio (1g:8mL). Then add a small amount of quartz sand and grind evenly to obtain a fine slurry. After that, use four layers of gauze to filter out the remaining solid particles and collect the filtered virus stock solution for later use.

[0039] The control group sprayed clean water on the tobacco plant leaves, and the treatment group sprayed the microbial agent of Example 1 diluted 50 times on the tobacco plant leaves. After 48 hours, the prepared cucumber mosaic virus (CMV) stock solution was used for friction inoculation. Ten plants were treated in each group, and 3 repetitions were set. The disease conditions of the plants were observed and recorded 20 days and 30 days after inoculation of cucumber mosaic virus (CMV), and the disease survey was conducted at the tobacco maturity stage to count the disease caused by cucumber mosaic virus (CMV): the morbidity of tobacco cucumber mosaic virus disease, and the disease index and relative control effect at different periods were calculated according to the grading standard.

[0040] According to the provisions on tobacco disease classification in the "Tobacco Industry Standard of the People's Republic of China YC / T 39-1996", especially the classification standards for tobacco mosaic disease, this standard adopts the method of using the whole plant as the evaluation unit.

[0041] Disease index = ∑ (disease level × number of diseased plants) / (highest disease level × total number of plants in each treatment) × 100

[0042] Control effect (%) = (control disease index - treatment disease index) / control disease index × 100%

[0043] In the pot test, during the growth and fertilization period after tobacco sowing, after the application of microbial agents and the inoculation of cucumber mosaic virus (CMV), the phenotypic results of tobacco plants in different treatment groups are as follows: Figure 1 The statistical results of tobacco disease index and prevention and control effects are shown in Table 1.

[0044] Table 1 Statistics of the number of tobacco plants infected with diseases

[0045]

[0046] As can be seen from Table 1, when the tobacco grew for 20 days, the incidence of cucumber mosaic virus disease in the control group was about 65.63%, while the incidence of the tobacco in the treatment group treated with the microbial agent was about 34.38%, which significantly reduced the incidence of diseases caused by CMV and had the effect of inhibiting CMV. When the tobacco grew for 30 days, the incidence of cucumber mosaic virus disease in the control group was about 93.75%, while the incidence of cucumber mosaic virus disease in the treatment group treated with the microbial agent was about 56.25%. It can be seen that the application of the microbial agent containing △PS04-17 has a significant preventive effect on tobacco cucumber mosaic virus disease, and the control effect on tobacco diseases caused by CMV in the field reached 54.72% when the tobacco grew for 20 days and 47.90% when the tobacco grew for 30 days. It shows that the space-induced mutant strain △PS04-17 of Paenibacillus Cliburn can well prevent the occurrence of tobacco cucumber mosaic virus disease, effectively inhibit the spread of CMV in the plant, delay the onset of the disease, and prevent premature aging of the host plant.

[0047] Application of Experimental Example 3 in Tobacco Growth

[0048] After sowing, the tobacco seeds are placed in a constant temperature culture environment for cultivation, and regular irrigation is required to maintain appropriate moisture. After the tobacco seedlings successfully break through the soil, transplantation is carried out. The experiment is divided into two groups, with 7 tobacco plants in each group. The control group sprays clean water on the leaves of the tobacco plants, and the treatment group sprays the tobacco leaves with a 100-fold dilution of microbial agents. Both are sprayed once every 7 days, for a total of two sprays. In addition to the difference in whether microbial agents are applied, the other treatments are the same. During the experiment, no other agents were used in each group to control pests and diseases. When the tobacco grows to the stage with 7 fully expanded leaves, the maximum leaf area, fresh weight of the aboveground part and the underground part of each group are measured.

[0049] The phenotypic results of tobacco plants in different treatment groups are as follows Figure 2 As shown in Figure 2, the average fresh weight of the aboveground part of each plant in the treatment group treated with microbial agents reached 3.5g, while the average fresh weight of each plant in the control group was 1.8g. The results were significantly different. Figure 3 As shown in the figure, the average fresh weight of the underground part of each plant in the treatment group reached 0.16g, while the average fresh weight of the underground part of each plant in the control group was 0.03g. The results were significantly different. The statistics are as follows Figure 4 As shown; the average maximum leaf area of ​​each plant in the treatment group reached 36cm 2 The maximum leaf area of ​​each plant in the control group was 22 cm 2 The results are significantly different, as shown in the statistics Figure 5 The results showed that during the tobacco growth period, spraying with a 100-fold diluted microbial agent of Paenibacillus sp. space mutant △PS04-17 strain can promote the development of tobacco roots, enhance the growth of tobacco, promote tobacco growth, increase tobacco leaf area, and increase tobacco yield.

[0050] In summary, the present invention provides a new application of the space-induced mutant strain △PS04-17 of Paenibacillus cribensis in the prevention and treatment of cucumber mosaic virus. Studies have shown that the △PS04-17 strain can significantly reduce the incidence of CMV host plants and has the effect of inhibiting CMV. After the host plant is treated with the fermentation liquid of △PS04-17, the relative prevention effect of tobacco cucumber mosaic virus disease caused by CMV reaches 54.72%, which can prevent the occurrence of tobacco cucumber mosaic virus disease, prevent premature aging of tobacco plants, effectively inhibit the spread of CMV in the plant body, and delay the onset of the disease; at the same time, the microbial agent prepared by the △PS04-17 strain can effectively promote the growth of tobacco plants, promote the development of tobacco roots, increase the area of ​​tobacco leaves, enhance the growth of tobacco, and have obvious effects of increasing production and quality. It can not only reduce the use of pesticides, but also be conducive to the development of environmental safety. It can be used for green prevention and control of tobacco, which is of great significance to the quality and efficiency of tobacco.

[0051] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. Application of the space mutant △PS04-17 strain of Paenibacillus kribbensis in the prevention and control of cucumber mosaic virus.

2. Application of the space mutant △PS04-17 strain of Bacillus kriebensis in preventing and controlling plant diseases caused by cucumber mosaic virus.

3. Application of the space mutant △PS04-17 strain of Bacillus cribenbergii in the preparation of cucumber mosaic virus control products.

4. Application of the space mutant △PS04-17 strain of Bacillus cribenbergii in promoting tobacco growth.

5. Application of the space mutant △PS04-17 strain of Bacillus cribenbergii in the preparation of growth promoters.

6. Application of the space mutant △PS04-17 strain of Bacillus cribenbergii in improving tobacco yield.

7. A method for preventing and controlling cucumber mosaic virus or reducing the incidence of cucumber mosaic virus disease, characterized in that: The plants were treated with the space mutant △PS04-17 strain of Paenibacillus kriebensis or its bacterial solution.

8. The method according to claim 7, characterized in that: The bacterial liquid is a fermentation liquid.

9. The method according to claim 8, characterized in that: The fermentation liquid concentration is not less than 1×10 9 CFU / mL.

10. The use according to any one of claims 1 to 6, or the method according to claim 7, characterized in that: The Bacillus Klibensis space mutant △PS04-17 strain was deposited in the Guangdong Provincial Microbial Culture Collection Center on March 10, 2022, with the deposit number: GDMCC NO: 62233.

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