Preparation and application method of synergistic additive for improving disease prevention effect of biocontrol bacillus on watermelon fusarium wilt

Abstract

The invention relates to a preparation method and an application method of a synergistic aid for improving the disease prevention effect of biocontrol bacillus on watermelon fusarium wilt, and belongs to the technical field of biocontrol synergistic aids. The formula of the synergistic auxiliary agent for improving the disease prevention effect of biocontrol bacillus on watermelon fusarium wilt comprises the following components in parts by weight: 40-50 parts of humic acid, 35-40 parts of sucrose, 10-15 parts of oxalic acid, 3-5 parts of lactic acid and 1-2 parts of spermidine. With the adoption of the synergistic auxiliary agent, the growth-promoting, disease-preventing and stress-resisting effects of the biocontrol bacillus on plants can be effectively improved, the promoting effect of the biocontrol bacillus on crop growth can be effectively improved, and the biomass, including the chlorophyll content, the vine length, the stem thickness and the leaf number of the plants, of the plants can be remarkably increased. And meanwhile, by adopting the synergistic auxiliary agent, the abiotic stress resistance of the biocontrol bacillus can be effectively enhanced, the disease severity of crop field diseases can be obviously reduced, and the disease prevention rate of the biocontrol bacillus on the crops can be obviously improved.

Description

Technical Field

[0001] The invention relates to the technical field of biocontrol synergistic adjuvants for microorganisms, and in particular to a preparation and application method of a synergistic adjuvant for improving the disease control effect of biocontrol Bacillus on watermelon wilt. Background Art

[0002] Bacillus is one of the most valuable biocontrol bacteria. It is gradually showing its advantages due to its easy cultivation, short reproduction cycle, and convenient production. In addition to being used as a microbial agent and microbial fertilizer additive, Bacillus can promote plant growth and increase production, improve plant immunity, and reduce plant diseases. However, as a live bacterial preparation, Bacillus is greatly affected by the environment in production and application, has a short survival period, and a single mode of action. These shortcomings limit the application effect and stability of microbial agents. Therefore, it is necessary to develop synergistic adjuvants that improve the colonization ability of Bacillus, enhance its growth-promoting and disease-prevention effects, and expand its mode of action.

[0003] Microbial biocontrol synergistic adjuvants refer to a class of microbial live bacteria preparations that can improve microbial performance and enhance the effect of microbial application. The main functions of microbial biocontrol synergistic adjuvants are to assist biocontrol bacteria in enhancing the induced plant systemic disease resistance, increase the activity of microbial cells, improve the colonization ability of biocontrol bacteria in the rhizosphere, and reduce the stress of adversity on biocontrol bacteria. Summary of the invention

[0004] The invention provides a synergistic auxiliary agent for improving the disease prevention effect of biocontrol bacillus on watermelon wilt. The synergistic auxiliary agent comprises the following raw materials in parts by weight: 40-50 parts of humic acid, 35-40 parts of sucrose, 10-15 parts of oxalic acid, 3-5 parts of lactic acid and 1-2 parts of spermidine.

[0005] The present invention provides a method for preparing the biocontrol synergistic adjuvant, comprising the following steps: (1) Dry humic acid, sucrose, oxalic acid, lactic acid, and spermidine at 40-45° C. for 5-6 hours, grind them, and pass them through a 60-mesh sieve to obtain a raw material; (2) weighing raw materials according to the weight ratio of 40-50 parts of humic acid, 35-40 parts of sucrose, 10-15 parts of oxalic acid, 3-5 parts of lactic acid, and 1-2 parts of spermidine, and mixing them thoroughly to obtain a mixture; (3) The mixture was stirred at 220 r / min for 100 min under 180 W ultrasonic conditions and dried at 45 ° C for 6 h to obtain the synergistic aid.

[0006] The present invention provides a method for applying a synergistic adjuvant for improving the disease prevention effect of biocontrol Bacillus on watermelon wilt, comprising the following steps: (1) When spraying on leaves, the application rate is 750g / hm2 , dilute it at a mass ratio of 1:200, mix it with the biocontrol Bacillus agent, and spray it evenly on the leaves; (2) When irrigating the roots, the application rate is 12 kg / hm 2 , first apply the synergistic adjuvant, then apply the biocontrol Bacillus agent. BRIEF DESCRIPTION OF THE DRAWINGS

[0007] Figure 1 This is a diagram of the effect of Bacillus synergistic adjuvant on enhancing the growth promotion effect of Bacillus on watermelon in the field. The clean water treatment was used as the control group.

[0008] Figure 2 This is a diagram of the effect of Bacillus synergistic adjuvant on enhancing the field control effect of Bacillus on watermelon wilt. The treatment with clean water was used as the control group. DETAILED DESCRIPTION

[0009] The present invention is explained below in conjunction with the embodiments, and the technical solutions in the embodiments of the present invention are clearly and completely described. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. The embodiments in the present invention are preferred implementation methods of the present invention, and any other changes, modifications, substitutions, combinations, and simplifications made without departing from the spirit and principle of the present invention shall be equivalent replacement methods and are included in the protection scope of the present invention. Example 1

[0010] The present embodiment provides a synergistic auxiliary agent for improving the disease prevention effect of biocontrol Bacillus on watermelon wilt. The synergistic auxiliary agent comprises the following raw materials in parts by weight: 40-50 parts of humic acid, 35-40 parts of sucrose, 10-15 parts of oxalic acid, 3-5 parts of lactic acid, and 1-2 parts of spermidine. Example 2

[0011] This embodiment provides a method for preparing the biocontrol synergistic adjuvant, comprising the following steps: (1) Dry humic acid, sucrose, oxalic acid, lactic acid, and spermidine at 40-45° C. for 5-6 hours, grind them, and pass them through a 60-mesh sieve to obtain a raw material; (2) weighing raw materials according to the weight ratio of 40-50 parts of humic acid, 35-40 parts of sucrose, 10-15 parts of oxalic acid, 3-5 parts of lactic acid, and 1-2 parts of spermidine, and mixing them thoroughly to obtain a mixture; (3) The mixture was stirred at 220 r / min for 100 min under 180 W ultrasonic conditions and dried at 45 ° C for 6 h to obtain the synergistic aid. Example 3

[0012] This embodiment provides a method for applying a synergistic adjuvant to improve the disease prevention effect of biocontrol Bacillus on watermelon wilt, comprising the following steps: (1) When spraying on leaves, the application rate is 750g / hm 2 , dilute it at a mass ratio of 1:200, mix it with the biocontrol Bacillus agent, and spray it evenly on the leaves; (2) When irrigating the roots, the application rate is 12 kg / hm 2 , first apply the synergistic adjuvant, then apply the biocontrol Bacillus agent. Example 4

[0013] This embodiment provides an application example of a synergistic adjuvant for promoting growth in watermelon fields: The test variety is Jingmei 1K, provided by Jingyan Yinong (Shouguang) Seed Technology Co., Ltd. The test fertilizer is a Bacillus agent, an aqueous solution, with an effective number of live bacteria (Bacillus Velez, Bacillus licheniformis) ≥ 1 billion / mL. The experiment was carried out in a greenhouse in Xiangshui County, Yancheng, Jiangsu. The experiment has four treatments, namely control (CK), Bacillus treatment, synergist treatment, and Bacillus + synergist treatment. Each treatment was repeated 3 times and arranged in random blocks. The plant spacing is 0.25 m, the row spacing is 0.8 m, and the plot area is 25m 2 , 60 plants were planted in the plot, and drainage and irrigation ditches were opened around each treatment area to ensure that drainage and irrigation were convenient. Except for the different fertilization treatments, the field management measures of each treatment were consistent, and fertilization and watering were carried out in an integrated manner.

[0014] On the day of transplanting watermelon seedlings, 20 ml of clean water was irrigated to each seedling as the control group. After diluting the Bacillus agent 50 times, 20 ml of water was irrigated to each seedling as the Bacillus treatment group. After diluting the synergist at a mass ratio of 1:20, 20 ml of water was irrigated to each seedling as the synergist treatment group. The synergist was first diluted at a mass ratio of 1:10, and the Bacillus agent was diluted 25 times. After equal volumes were mixed evenly, 20 ml of water was irrigated to each seedling as the Bacillus + synergist treatment group. After 15 days of treatment, the changes in the crop seedlings were observed. The effect can be seen in the Figure 1 . See Figure 1 , which is a diagram showing the effect of Bacillus synergistic adjuvant on enhancing the growth promotion of Bacillus on watermelon in the field. The treatment with clean water is marked as the "control group", the treatment with synergistic adjuvant is marked as the "synergistic adjuvant treatment group", the treatment with Bacillus is marked as the "Bacillus treatment group", and the treatment with synergistic adjuvant and Bacillus is marked as the "Bacillus + synergistic adjuvant treatment group".

[0015] The biomass of plants in each treatment was counted: stem diameter, number of leaves, plant height, and chlorophyll.

[0016] The statistical results show that compared with the control group, the synergist treatment group and the Bacillus treatment group have significantly increased various biomasses (chlorophyll, vine length, stem diameter, and leaf number), and compared with the synergist treatment group and the Bacillus treatment group, the Bacillus + synergist treatment group has significantly increased various biomasses (chlorophyll, vine length, stem diameter, and leaf number). Therefore, the results show that the synergist of the present invention can significantly enhance the growth-promoting effect of Bacillus. Example 5

[0017] This embodiment provides an application example of a synergistic adjuvant in the prevention and treatment of watermelon field wilt: The test variety is Jingmei 1K, provided by Jingyan Yinong (Shouguang) Seed Technology Co., Ltd. The test fertilizer is a Bacillus agent, an aqueous solution, with an effective number of live bacteria (Bacillus Velez, Bacillus licheniformis) ≥ 1 billion / mL. The experiment was carried out in a multi-span greenhouse in Xiangshui County, Yancheng, Jiangsu. The selected plot is a three-year continuous crop of watermelon. The previous crop was watermelon, and watermelon wilt disease occurred evenly in the plot, with an incidence rate of more than 80%. The experiment has four treatments, namely control (CK), Bacillus treatment, synergist treatment, and Bacillus + synergist treatment. Each treatment was repeated 3 times and arranged in random blocks. The plant spacing is 0.25 m, the row spacing is 0.8 m, and the plot area is 25m 2 , 60 plants were planted in the plot, and drainage and irrigation ditches were opened around each treatment area to ensure that drainage and irrigation were convenient. Except for the different fertilization treatments, the field management measures of each treatment were consistent, and fertilization and watering were carried out in an integrated manner.

[0018] On the day of transplanting watermelon seedlings, irrigate each seedling with 20 ml of clean water as the control group. Dilute the Bacillus agent 50 times and irrigate each seedling with 20 ml of root water as the Bacillus treatment group. Dilute the synergist 20 times and irrigate each seedling with 20 ml of root water as the synergist treatment group. First dilute the synergist 10 times and the Bacillus agent 25 times, mix them evenly in equal volumes, and irrigate each seedling with 20 ml of root water as the Bacillus + synergist treatment group. After the watermelon enters the fruiting period, spray it every 10 days to prevent leaf diseases. The control group was sprayed with water every 10 days; the Bacillus treatment group was sprayed every 10 days after the Bacillus agent was diluted 200 times; the synergist treatment group was sprayed every 10 days after the synergist was diluted at a mass ratio of 1:200; the Bacillus + synergist treatment group was first diluted at a mass ratio of 1:10, and the Bacillus agent was diluted 25 times, and then sprayed every 10 days after equal volume was mixed. After 70 days of treatment, the occurrence of watermelon wilt was observed. The effect is shown in Figure 2 . See Figure 2, which is a diagram showing the effect of Bacillus synergistic adjuvant on enhancing the field prevention effect of Bacillus on watermelon wilt. The treatment with clean water is marked as the "control group", the treatment with synergistic adjuvant is marked as the "synergistic adjuvant treatment group", the treatment with Bacillus is marked as the "Bacillus treatment group", and the treatment with synergistic adjuvant and Bacillus is marked as the "Bacillus + synergistic adjuvant treatment group".

[0019] Investigate and record the incidence of watermelon wilt in each treatment, and calculate the disease index, incidence rate and control effect. The disease classification is divided into 6 levels, namely: Level 0: watermelon plants have no symptoms; Level 1: watermelon plants grow well overall, the cotyledons are yellowed and wrinkled, and the base of the stem turns slightly yellow; Level 2: watermelon plants have wilted cotyledons, the base of the stem turns yellow-brown, the stem is good and can grow upright; Level 3: watermelon plants have obvious wilting cotyledons, and the stem turns brown and wilts moderately; Level 4: watermelon plants are severely wilted, the cotyledons and stems are obviously wilted, and the stems show obvious browning symptoms; Level 5: the whole watermelon plant wilts, dies or emerges late, and dies or grows hyphae after emergence. Disease index = Σ[(each disease level × the number of diseased plants at each level) / (total number of plants × the highest disease level)] × 100. Control effect = (control disease index - treatment disease index) / control disease index × 100%.

[0020] The statistical results show that compared with the control group, the synergist treatment group, the bacillus treatment group, and the bacillus + synergist treatment group, the disease severity is significantly reduced, among which the bacillus + synergist treatment group has the lowest disease severity and a higher control effect on watermelon wilt, reaching 82.48% compared with the control group. Compared with the synergist treatment group and the bacillus treatment group, the disease severity of the bacillus + synergist treatment group is also significantly reduced. Therefore, the results show that the synergist of the present invention can significantly enhance the control effect of bacillus on watermelon wilt.

[0021] The series of detailed descriptions listed above are only specific descriptions of feasible implementation methods of the present invention. They are not intended to limit the scope of protection of the present invention. Any equivalent implementation methods or changes that do not deviate from the technical spirit of the present invention should be included in the scope of protection of the present invention.

Claims

1. A synergistic adjuvant for improving the disease prevention effect of biocontrol Bacillus on watermelon wilt, characterized in that: The invention contains the following components in parts by weight: 40-50 parts of humic acid, 35-40 parts of sucrose, 10-15 parts of oxalic acid, 3-5 parts of lactic acid and 1-2 parts of spermidine.

2. A synergistic adjuvant for improving the disease prevention effect of biocontrol Bacillus against watermelon wilt according to claim 1, characterized in that: Humic acid is in the form of potassium salt and its content is 65%-70%, and fulvic acid is 15%-20%.

3. A method for preparing a synergistic adjuvant for improving the disease prevention effect of biocontrol Bacillus on watermelon wilt according to claim 1, characterized in that: The following steps are involved: (1) humic acid, sucrose, oxalic acid, lactic acid, and spermidine are dried and crushed respectively, and passed through a 60-mesh sieve to obtain a raw material; (2) weighing raw materials according to the weight ratio of 40-50 parts of humic acid, 35-40 parts of sucrose, 10-15 parts of oxalic acid, 3-5 parts of lactic acid, and 1-2 parts of spermidine, and mixing them thoroughly to obtain a mixture; (3) The mixture is stirred under ultrasonic conditions and dried to obtain the synergistic agent.

4. The method for preparing a synergistic adjuvant for improving the disease prevention effect of biocontrol Bacillus on watermelon wilt according to claim 3, characterized in that: The drying temperature in steps (1) and (3) is 40-45° C. and the drying time is 5-6 hours.

5. The method for preparing a synergistic adjuvant for improving the disease prevention effect of biocontrol Bacillus on watermelon wilt according to claim 3, characterized in that: In step (3), the ultrasonic power is 180-200 W, the stirring speed is 200-220 r / min, and the stirring time is 1.5-2 h.

6. A synergistic adjuvant for improving the disease prevention effect of biocontrol Bacillus on watermelon wilt according to claim 1, wherein the application method of the synergistic adjuvant for improving the disease prevention effect of biocontrol Bacillus on watermelon wilt is characterized in that: The following steps are involved: (1) When spraying on leaves, the application rate is 750g / hm 2 , dilute it at a mass ratio of 1:200, mix it with the biocontrol Bacillus agent, and spray it evenly on the leaves; (2) When irrigating the roots, the application rate is 12 kg / hm 2 , first apply the synergistic adjuvant, then apply the biocontrol Bacillus agent.

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