Method for preventing and treating tomato bacterial wilt

Abstract

The invention discloses a tomato bacterial wilt prevention and control method, which belongs to the technical field of tomato disease prevention and control, and comprises the following steps: S1, preparing a complex microbial inoculant: in the complex microbial inoculant, the volume ratio of bacillus velezensis bacterial liquid to stenotrophomonas rhizophila bacterial liquid to streptomyces griseorubens bacterial liquid is 1: (3-6): (2-4); the effective viable count of the bacillus riissii bacterial liquid, the stenotrophomonas rhizophila bacterial liquid and the streptomyces griseorubens bacterial liquid is not less than 1 * 10 < 8 > CFU / ml; s2, applying the complex microbial inoculant obtained in the step S1 to rhizosphere of tomato plants. The method provided by the invention can be used for effectively preventing and treating the tomato bacterial wilt caused by the Ralstonia solanacearum (Ralstonia solanacearum).

Description

Technical Field

[0001] This invention belongs to the field of tomato disease control technology, specifically, it relates to a method for controlling bacterial wilt of tomatoes. Background Technology

[0002] Bacterial wilt of tomatoes is a soil-borne bacterial disease. The pathogen is *Ralstonia solanaceae*. (Ralstonia) solanacearum) It can survive in the soil for a long time, invade through root wounds, colonize the vascular bundles, causing the plant to wilt and die rapidly, resulting in reduced yield and affecting tomato cultivation. Summary of the Invention

[0003] In order to overcome the problems existing in the prior art, the present invention proposes a method for the prevention and control of bacterial wilt in tomatoes.

[0004] To achieve the above objectives, the present invention is implemented through the following technical solution: A method for controlling bacterial wilt in tomatoes includes the following steps: S1, Preparation of compound bacterial agent: The compound bacterial agent contains Bacillus belye ( Bacillus velezensis ) bacterial solution, rhizotrophic oligotrophosomes ( Stenotrophomonas rhizophila ) bacterial solution and Streptomyces griseus ( Streptomyces griseorubens The volume ratio of the bacterial suspensions was 1:3-6:2-4; the effective viable counts of the *Bacillus repens*, *Oligotrophomonas radiata*, and *Streptomyces griseus* suspensions were all not less than 1×10⁻⁶. 8 CFU / ml; S2, apply the compound microbial agent obtained in step S1 to the rhizosphere of the tomato plants.

[0005] Furthermore, the method for preparing the Bacillus belyss bacterial culture includes the following steps: 1) The liquid culture was cultured using LB liquid medium at 28℃ and 180 r / min for 2 days to obtain the liquid culture. The culture medium formula was: 10 g / L tryptone, 5 g / L yeast extract, 5 g / L NaCl, pH=7.0. 2) The liquid bacterial strain obtained in step 1) was inoculated into the liquid fermentation medium at an inoculation rate of 3%, and cultured at 30℃ and 220r / min for 3 days. The fermentation broth was then filtered, and the filtrate was diluted to obtain Bacillus belyssus culture. The liquid fermentation medium formula was: 10g / L tryptone, 5g / L yeast extract, 5g / L NaCl, pH=7.0.

[0006] Furthermore, the method for preparing the *Oligotrophomonas rhizophila* bacterial suspension includes the following steps: A1. Preparation of liquid culture: *Oligotrophomonas radiata* was inoculated into a liquid culture medium and cultured at 30°C and 180 r / min for 24-36 h to obtain a liquid culture; the liquid culture medium contained: peptone: 10.0 g / L, beef meal: 3.0 g / L, sodium chloride: 5.0 g / L, pH=7.3; A2, the liquid bacterial strain obtained in step A1 is inoculated into the liquid fermentation medium at an inoculum rate of 3%, and cultured at 30℃ and 220r / min for 3 days. The fermentation broth is then filtered, and the filtrate is diluted to obtain the root-loving oligotrophomonas bacterial suspension. The liquid fermentation medium contains: peptone: 10.0g / L, beef meal: 3.0g / L, sodium chloride: 5.0g / L, pH=7.3.

[0007] Furthermore, the method for preparing the *Streptomyces griseus* bacterial suspension includes the following steps: B1. Preparation of liquid culture: *Streptomyces griseus* was inoculated into a liquid culture medium and cultured at 32℃ and 200 r / min for 3 days to obtain a liquid culture. The liquid culture medium contained: 20.0 g / L soluble starch, 1.0 g / L KNO3, 0.5 g / L K2HPO4, 0.5 g / L MgSO4·7H2O, 0.5 g / L NaCl, pH = 7.2. B2. The liquid inoculum obtained in step B1 is inoculated into the liquid fermentation medium at an inoculum volume of 3%, and cultured at 32℃ and 220 r / min for 5 days. The fermentation broth is then filtered, and the filtrate is diluted to obtain the *Streptomyces griseus* bacterial culture. The liquid fermentation medium contains: 20.0 g / L soluble starch, 1.0 g / L KNO3, 0.5 g / L K2HPO4, 0.5 g / L MgSO4·7H2O, 0.5 g / L NaCl, and pH = 7.2.

[0008] Furthermore, in step S2, the amount of compound bacterial solution applied is 200-300 ml / strain.

[0009] Through the above technical solution, the present invention can achieve at least the following beneficial effects: The method described in this invention can effectively prevent and control diseases caused by Rhesus solani (Solanaceae). (Ralstonia solanacearum) This causes bacterial wilt in tomatoes. Detailed Implementation

[0010] Unless otherwise stated, all materials and reagents used in this invention are commercially available.

[0011] In this invention, *Oligotrophomonas rhizophila*, *Streptomyces griseus*, and *Bacillus bellis* were all purchased from Beijing Bio-Biobio Biotechnology Co., Ltd. The strain number of *Oligotrophomonas rhizophila* is Bio-099392, the strain number of *Streptomyces griseus* is Bio-01152, and the strain number of *Bacillus bellis* is Bio-85113.

[0012] The method for preparing the *Oligotrophomonas rhizophila* bacterial suspension in this invention includes the following steps: A1. Preparation of liquid culture: *Oligotrophomonas radiata* was inoculated into a liquid culture medium and cultured at 30°C and 180 r / min for 24-36 h to obtain a liquid culture; the liquid culture medium contained: peptone: 10.0 g / L, beef meal: 3.0 g / L, sodium chloride: 5.0 g / L, pH=7.3; A2, the liquid inoculum obtained in step A1 is inoculated into the liquid fermentation medium at an inoculum rate of 3%, and cultured at 30℃ and 220 r / min for 3 days. After filtration, the fermentation broth is diluted to obtain an effective viable count of approximately 1×10⁻⁶. 8 The liquid fermentation medium contains CFU / ml of *Oligotrophomonas radiata* culture medium, which includes: peptone: 10.0 g / L, beef meal: 3.0 g / L, sodium chloride: 5.0 g / L, pH=7.3.

[0013] The method for preparing Bacillus belyssus culture in this invention includes the following steps: 1) The liquid culture was cultured using LB liquid medium at 28℃ and 180 r / min for 2 days to obtain the liquid culture. The culture medium formula was: 10 g / L tryptone, 5 g / L yeast extract, 5 g / L NaCl, pH=7.0. 2) Inoculate the liquid bacterial culture obtained in step 1) into the liquid fermentation medium at an inoculum rate of 3%, and incubate at 30℃ and 220 r / min for 3 days. After filtration, dilute the filtrate to obtain an effective viable count of approximately 1×10⁻⁶ cells / day. 8 The liquid fermentation medium consisted of CFU / ml Bacillus belyssus culture medium, which was prepared as follows: 10 g / L tryptone, 5 g / L yeast extract, 5 g / L NaCl, pH=7.0.

[0014] The method for preparing *Streptomyces griseus* bacterial suspension in this invention includes the following steps: B1. Preparation of liquid culture: *Streptomyces griseus* was inoculated into a liquid culture medium and cultured at 32℃ and 200 r / min for 3 days to obtain a liquid culture. The liquid culture medium contained: 20.0 g / L soluble starch, 1.0 g / L KNO3, 0.5 g / L K2HPO4, 0.5 g / L MgSO4·7H2O, 0.5 g / L NaCl, pH = 7.2. B2. The liquid inoculum obtained in step B1 was inoculated into the liquid fermentation medium at an inoculum rate of 3%. After incubation at 32℃ and 220 rpm for 5 days, the fermentation broth was filtered, and the filtrate was then diluted to obtain an effective viable count of approximately 1 × 10⁻⁶ cells / day. 8 The liquid fermentation medium contains CFU / ml of *Streptomyces griseus* culture medium, which includes 20.0 g / L soluble starch, 1.0 g / L KNO3, 0.5 g / L K2HPO4, 0.5 g / L MgSO4·7H2O, 0.5 g / L NaCl, and pH = 7.2.

[0015] Prepare the compound microbial agent according to the proportions shown in Table 1 below:

[0016] Tomato seedlings were transplanted into flowerpots and then grown in greenhouses for one month. 1400 healthy tomato plants with no obvious diseases and similar growth were selected for the experiment and divided into six treatment groups (AF) and one control group, with 200 plants in each group.

[0017] The six treatment groups of tomato plants were treated with root irrigation using compound microbial agents numbered 1-6, respectively. Compound microbial agent No. 1 was used to drench the roots of tomato plants in Group A, 200 ml per plant; The compound microbial agent No. 2 was used to drench the roots of the tomato plants in Group B, 250 ml per plant. Compound microbial agent No. 3 was used to drench the roots of tomato plants in group C, 300 ml per plant; Compound microbial agent No. 4 was used to drench the roots of tomato plants in group D, 200 ml per plant; Compound microbial agent No. 5 was used to drench the roots of tomato plants in group E, 200 ml per plant; Compound microbial agent No. 6 was used to drench the roots of tomato plants in group F, 200 ml per plant; The control group was treated with 200ml of sterile water for root irrigation.

[0018] Fifteen days after the above root irrigation treatment, use an effective viable bacteria count of 1×10⁶. 8 The root irrigation treatment of the AF group and the control group was carried out with a CFU / ml Rhesus solani suspension, with 200ml of the suspension applied to each tomato plant.

[0019] The rest of the management was normal. One month after the root irrigation with Ralstonia solanacearum suspension, the incidence of bacterial wilt was investigated. Incidence rate = number of infected plants / total number of plants surveyed × 100%.

[0020] The statistical results are shown in Table 2 below: serial number Number of infected plants / plant Incidence rate / % A 4 2 B 2 1 C 2 1 D 68 34 E 87 43.5 F 71 30.5 control group 183 91.5 As can be seen from Table 2 above, the compound microbial agent of this application can effectively prevent and control diseases caused by Rhesus solani. (Ralstonia) solanacearum) This causes bacterial wilt in tomatoes.

[0021] Finally, it should be noted that the above preferred embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail through the above preferred embodiments, those skilled in the art should understand that various changes can be made to it in form and detail without departing from the scope defined by the claims of the present invention.

Claims

1. A method for controlling bacterial wilt of tomatoes, characterized in that: It includes the following steps: S1, Preparation of compound bacterial agent: The compound bacterial agent contains Bacillus belye ( Bacillus velezensis ) bacterial suspension, rhizotrophic oligotrophosomes ( Stenotrophomonas rhizophila ) bacterial solution and Streptomyces griseus ( Streptomyces griseorubens The volume ratio of the bacterial suspensions was 1:3-6:2-4; the effective viable counts of the *Bacillus repens*, *Oligotrophomonas radiata*, and *Streptomyces griseus* suspensions were all not less than 1×10⁻⁶. 8 CFU / ml; S2, apply the compound microbial agent obtained in step S1 to the rhizosphere of the tomato plants.

2. The method for controlling bacterial wilt of tomatoes according to claim 1, characterized in that: The method for preparing the Bacillus vesiculosus culture includes the following steps: The liquid culture was cultured in LB liquid medium at 28℃ and 180 rpm for 2 days to obtain the liquid culture. The culture medium formula was: 10 g / L tryptone, 5 g / L yeast extract, 5 g / L NaCl, pH=7.

0. The liquid bacterial strain obtained in step 1) was inoculated into the liquid fermentation medium at an inoculation rate of 3%, and cultured at 30℃ and 220r / min for 3 days. The fermentation broth was then filtered, and the filtrate was diluted to obtain Bacillus belyssus culture. The liquid fermentation medium formula was: 10g / L tryptone, 5g / L yeast extract, 5g / L NaCl, pH=7.

0.

3. The method for controlling bacterial wilt of tomatoes according to claim 1, characterized in that: The method for preparing the rhizotrophic oligotrophomonas bacterial suspension includes the following steps: A1. Preparation of liquid culture: *Oligotrophomonas radiata* was inoculated into a liquid culture medium and cultured at 30°C and 180 r / min for 24-36 h to obtain a liquid culture; the liquid culture medium contained: peptone: 10.0 g / L, beef meal: 3.0 g / L, sodium chloride: 5.0 g / L, pH=7.3; A2, the liquid bacterial strain obtained in step A1 is inoculated into the liquid fermentation medium at an inoculum rate of 3%, and cultured at 30℃ and 220r / min for 3 days. The fermentation broth is then filtered, and the filtrate is diluted to obtain the root-loving oligotrophomonas bacterial suspension. The liquid fermentation medium contains: peptone: 10.0g / L, beef meal: 3.0g / L, sodium chloride: 5.0g / L, pH=7.

3.

4. The method for controlling bacterial wilt of tomatoes according to claim 1, characterized in that: The preparation method of the *Streptomyces griseus* bacterial suspension includes the following steps: B1. Preparation of liquid culture: *Streptomyces griseus* was inoculated into a liquid culture medium and cultured at 32℃ and 200 r / min for 3 days to obtain a liquid culture. The liquid culture medium contained: 20.0 g / L soluble starch, 1.0 g / L KNO3, 0.5 g / L K2HPO4, 0.5 g / L MgSO4·7H2O, 0.5 g / L NaCl, pH = 7.

2. B2. The liquid inoculum obtained in step B1 is inoculated into the liquid fermentation medium at an inoculum volume of 3%, and cultured at 32℃ and 220 r / min for 5 days. The fermentation broth is then filtered, and the filtrate is diluted to obtain the *Streptomyces griseus* bacterial culture. The liquid fermentation medium contains: 20.0 g / L soluble starch, 1.0 g / L KNO3, 0.5 g / L K2HPO4, 0.5 g / L MgSO4·7H2O, 0.5 g / L NaCl, and pH = 7.

2.

5. The method for controlling bacterial wilt of tomatoes according to claim 1, characterized in that: In step S2, the amount of compound microbial agent applied is 200-300 ml / strain.