A method for preventing tomato gray mold disease

Abstract

This invention discloses a method for controlling gray mold in tomatoes, belonging to the field of biotechnology, comprising the following steps: S1, preparing a compound microbial agent: the compound microbial agent contains deep red yeast (in a volume ratio of 1:3-6) Rhodotorula Rubra ) inoculants and lemon-shaped Klerk yeast ( Kloeckera Apiculata The inoculant, wherein the effective viable count of the deep red yeast inoculant is not less than 1×10⁻⁶. 8 CFU / ml, the effective viable count in the lemon-shaped Klebsiella pneumoniae agent is not less than 2×10⁻⁶ CFU / ml. 7 CFU / ml; S2, spray the compound microbial agent from step S1 onto the tomato plants. The tomato gray mold control method described in this invention can effectively control the fungus causing tomato gray mold. Botrytis cinerea This causes gray mold disease in tomatoes.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology, specifically, it relates to a method for controlling gray mold in tomatoes. Background Technology

[0002] Gray mold is a common and serious disease in tomato production. This pathogen can infect multiple parts of the tomato plant, forming "V"-shaped or circular spots on the leaves, causing leaf drop, and can also cause stem rot.

[0003] Currently, the main methods for controlling tomato gray mold include breeding resistant varieties, using chemical agents, and biological control. Due to the long breeding process and high difficulty, there are currently few resistant varieties. Using chemical agents can lead to environmental pollution and the development of drug resistance in the pathogen. Biological control methods utilize biocontrol bacteria selected from nature, making them environmentally friendly and safe for humans. Therefore, biological control has become a new method and a research hotspot in disease control. Summary of the Invention

[0004] To overcome the problems existing in the prior art, this invention proposes a method for controlling tomato gray mold. The method described in this invention can effectively control the fungus causing tomato gray mold. Botrytis cinerea This causes gray mold disease in tomatoes.

[0005] To achieve the above objectives, the present invention provides a method for controlling gray mold in tomatoes, comprising the following steps:

[0006] S1, Preparation of compound microbial agent: The compound microbial agent contains deep red yeast in a volume ratio of 1:3-6 ( Rhodotorula Rubra ) inoculants and lemon-shaped Klerk yeast ( Kloeckera Apiculata The inoculant, wherein the effective viable count of the deep red yeast inoculant is not less than 1×10⁻⁶. 8 CFU / ml, the effective viable count in the lemon-shaped Klebsiella pneumoniae agent is not less than 2×10⁻⁶ CFU / ml. 7 CFU / ml;

[0007] S2, spray the compound microbial agent from step S1 onto the tomato plants.

[0008] Furthermore, the preparation method of the deep red yeast agent includes the following steps:

[0009] A1. After expanding the purchased deep red yeast culture in test tubes, it was inoculated into liquid seed culture medium and cultured at 26℃-30℃, 180-200 r / min for 36-48 h to obtain liquid culture. The liquid seed culture medium contained the following components: yeast extract: 3.0 g / L, malt extract: 3.0 g / L, glucose: 10.0 g / L, casein peptone: 5.0 g / L, pH: 6.2±0.2.

[0010] A2, inoculate the liquid culture medium with 3%-5% of the inoculum, and ferment at 26℃-30℃, 180-220 r / min for 48-60 h. After filtration, obtain the fermentation broth and dilute it to 1×10⁻⁶. 8 CFU / ml was used to obtain the deep red yeast inoculum; the composition of the liquid fermentation medium was the same as that of the liquid seed medium in step A1.

[0011] Furthermore, the preparation method of the lemon-shaped Klerk yeast agent includes the following steps:

[0012] B1. After expanding the purchased *Klebsiella lemonata* test tube culture, the culture was inoculated into liquid seed culture medium and cultured at 25℃-28℃, 180-200 r / min, for 36-48 h to obtain *Klebsiella lemonata* liquid culture. The liquid seed culture medium contained the following components: 8 g / L beef extract, 5 g / L yeast powder, 10 g / L glucose, and natural pH.

[0013] B2: Inoculate the liquid culture medium with 1%-3% of the inoculum, and ferment at 26℃-30℃, 200-220 r / min for 60-72 h. After filtration, obtain the fermentation broth and dilute it to 2×10⁻⁶. 7 CFU / ml was used to obtain lemon-shaped Klerk yeast inoculum; the composition of the liquid fermentation medium was the same as that of the liquid seed medium in step B1.

[0014] Furthermore, the compound microbial agent contains deep red yeast ( Rhodotorula Rubra ) inoculants and lemon-shaped Klerk yeast ( Kloeckera Apiculata The volume ratio of the bacterial agent is 1:4.

[0015] Furthermore, the compound microbial agent contains deep red yeast ( Rhodotorula Rubra ) inoculants and lemon-shaped Klerk yeast ( Kloeckera Apiculata The volume ratio of the bacterial agent is 1:3.

[0016] Furthermore, the compound microbial agent contains deep red yeast ( Rhodotorula Rubra ) inoculants and lemon-shaped Klerk yeast ( Kloeckera Apiculata The volume ratio of the bacterial agent is 1:6.

[0017] Through the above technical solution, the present invention can achieve at least the following beneficial effects: the tomato gray mold control method described in the present invention can effectively control the disease caused by tomato gray mold fungus. Botrytis cinerea This causes gray mold disease in tomatoes. Detailed Implementation

[0018] Unless otherwise stated, all materials and reagents used in this invention are commercially available.

[0019] In this invention, both the deep red yeast and the lemon-shaped Klerk yeast were purchased from Beijing Bio-Bio Biotechnology Co., Ltd., with the deep red yeast having the serial number Bio-06784 and the lemon-shaped Klerk yeast having the serial number Bio-07302.

[0020] The preparation method of the deep red yeast agent in this invention includes the following steps:

[0021] A1. After expanding the purchased deep red yeast culture in test tubes, it was inoculated into liquid seed culture medium and cultured at 26℃-30℃, 180-200 r / min for 36-48 h to obtain liquid culture. The liquid seed culture medium contained the following components: yeast extract: 3.0 g / L, malt extract: 3.0 g / L, glucose: 10.0 g / L, casein peptone: 5.0 g / L, pH: 6.2±0.2.

[0022] A2, inoculate the liquid culture medium with 3%-5% of the inoculum, and ferment at 26℃-30℃, 180-220 r / min for 48-60 h. After filtration, obtain the fermentation broth and dilute it to 1×10⁻⁶. 8 CFU / ml was used to obtain the deep red yeast inoculum; the composition of the liquid fermentation medium was the same as that of the liquid seed medium in step A1.

[0023] The preparation method of lemon-shaped Klerk yeast agent in this invention includes the following steps:

[0024] B1. After expanding the purchased *Klebsiella lemonata* test tube culture, the culture was inoculated into liquid seed culture medium and cultured at 25℃-28℃, 180-200 r / min, for 36-48 h to obtain *Klebsiella lemonata* liquid culture. The liquid seed culture medium contained the following components: 8 g / L beef extract, 5 g / L yeast powder, 10 g / L glucose, and natural pH.

[0025] B2: Inoculate the liquid culture medium with 1%-3% of the inoculum, and ferment at 26℃-30℃, 200-220 r / min for 60-72 h. After filtration, obtain the fermentation broth and dilute it to 2×10⁻⁶. 7CFU / ml was used to obtain lemon-shaped Klerk yeast inoculum; the composition of the liquid fermentation medium was the same as that of the liquid seed medium in step B1. Example 1

[0026] A method for controlling tomato gray mold includes the following steps:

[0027] S1, Preparation of compound microbial agent: The compound microbial agent contains deep red yeast in a volume ratio of 1:4 ( Rhodotorula Rubra ) inoculants and lemon-shaped Klerk yeast ( Kloeckera Apiculata The inoculant, wherein the effective viable count of the deep red yeast inoculant is 1×10⁻⁶. 8 CFU / ml, the effective viable count in the lemon-shaped Klebsiella pneumoniae agent is 2×10⁻⁶ CFU / ml. 7 CFU / ml;

[0028] S2, spray the compound microbial agent from step S1 onto the tomato plants. Example 2

[0029] A method for controlling tomato gray mold includes the following steps:

[0030] S1, Preparation of compound microbial agent: The compound microbial agent contains deep red yeast in a volume ratio of 1:3 ( Rhodotorula Rubra ) inoculants and lemon-shaped Klerk yeast ( Kloeckera Apiculata The inoculant, wherein the effective viable count of the deep red yeast inoculant is 1×10⁻⁶. 8 CFU / ml, the effective viable count in the lemon-shaped Klebsiella pneumoniae agent is 2×10⁻⁶ CFU / ml. 7 CFU / ml;

[0031] S2, spray the compound microbial agent from step S1 onto the tomato plants. Example 3

[0032] A method for controlling tomato gray mold includes the following steps:

[0033] S1, Preparation of compound microbial agent: The compound microbial agent contains deep red yeast in a volume ratio of 1:6 ( Rhodotorula Rubra ) inoculants and lemon-shaped Klerk yeast ( Kloeckera Apiculata The inoculant, wherein the effective viable count of the deep red yeast inoculant is 1×10⁻⁶. 8 CFU / ml, the effective viable count in the lemon-shaped Klebsiella pneumoniae agent is 2×10⁻⁶ CFU / ml. 7 CFU / ml;

[0034] S2, spray the compound microbial agent from step S1 onto the tomato plants. Comparative Example 1

[0035] The difference between Comparative Example 1 and Example 1 is that the compound microbial agent does not contain deep red yeast ( Rhodotorula Rubra ) bacterial agent. Comparative Example 2

[0036] The difference between Comparative Example 2 and Example 1 is that the compound bacterial agent does not contain *Klerk's lemon-shaped yeast* (…). Kloeckera Apiculata ) bacterial agent. Experimental Example

[0037] Tomato seedlings aged 30 days, with uniform growth and no obvious diseases were selected for the experiment. First, the tomato leaves were sprayed with a compound microbial agent from Examples 1-3 and Comparative Examples 1-2. 24 hours later, a 1×10⁻⁶ concentration was sprayed. 6 Tomato gray mold (cacti / ml) Botrytis cinerea Spore suspension;

[0038] A control group was set up, with a direct spray concentration of 1×10⁻⁶. 6 Tomato gray mold (cacti / ml) Botrytis cinerea Spore suspension;

[0039] Examples 1 to 3, Comparative Examples 1 to 2, and the control group were all implemented in three replicates. Each treatment involved one tomato plant with five true leaves per plant.

[0040] The control group was surveyed and the incidence was recorded on the third day after the onset of the disease. The statistical results are shown in Table 1 below.

[0041] Disease severity grading criteria:

[0042] Grade 0, no lesions;

[0043] Grade 1: The lesion area accounts for less than 1 / 20 of the leaf area;

[0044] Grade 2, with lesions covering more than 1 / 20 to 1 / 6 of the leaf area;

[0045] Grade 3, with lesions covering more than 1 / 6 to 1 / 3 of the leaf area;

[0046] Level 4, with lesions covering more than 1 / 3 to 1 / 2 of the leaf area;

[0047] Level 5 disease covers more than half of the leaf area.

[0048] Disease index = ∑ (number of diseased leaves at each level × relative level value) / (total number of leaves surveyed × highest level value) × 100;

[0049] Prevention and control effect = (disease index of control group - disease index of treatment group) / disease index of control group × 100%.

[0050] Table 1. Control effects of different treatments on tomato gray mold.

[0051] deal with Disease index Prevention and control efficacy / % Example 1 5.31 93.77 Example 2 6.25 91.29 Example 3 5.28 94.02 Comparative Example 1 22.59 53.40 Comparative Example 2 24.71 47.81 CK 57.84 -

[0052] As can be seen from Table 1 above, the compound microbial agent of this application can effectively control tomato gray mold. Botrytis cinerea This leads to gray mold disease in tomatoes.

[0053] 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 gray mold in tomatoes, characterized in that: It includes the following steps: S1, Preparation of compound microbial agent: The compound microbial agent contains deep red yeast in a volume ratio of 1:3-6 ( Rhodotorula Rubra ) inoculants and lemon-shaped Klerk yeast ( Kloeckera Apiculata The inoculant, wherein the effective viable count of the deep red yeast inoculant is not less than 1×10⁻⁶. 8 CFU / ml, the effective viable count in the lemon-shaped Klebsiella pneumoniae agent is not less than 2×10⁻⁶ CFU / ml. 7 CFU / ml; S2, spray the compound microbial agent from step S1 onto the tomato plants.

2. The method for controlling tomato gray mold according to claim 1, characterized in that: The preparation method of the deep red yeast agent includes the following steps: A1. After expanding the purchased deep red yeast culture in test tubes, it was inoculated into liquid seed culture medium and cultured at 26℃-30℃, 180-200 r / min for 36-48 h to obtain liquid culture. The liquid seed culture medium contained the following components: yeast extract: 3.0 g / L, malt extract: 3.0 g / L, glucose: 10.0 g / L, casein peptone: 5.0 g / L, pH: 6.2±0.

2. A2, inoculate the liquid culture medium with 3%-5% of the inoculum, and ferment at 26℃-30℃, 180-220 r / min for 48-60 h. After filtration, obtain the fermentation broth and dilute it to 1×10⁻⁶. 8 CFU / ml was used to obtain the deep red yeast inoculum; the composition of the liquid fermentation medium was the same as that of the liquid seed medium in step A1.

3. The method for controlling tomato gray mold according to claim 1, characterized in that: The preparation method of the lemon-shaped Klerk yeast inoculum includes the following steps: B1. After expanding the purchased Klebsiella lemongrass test tube culture, it was inoculated into liquid seed culture medium and cultured at 25℃-28℃, 180-200 r / min for 36-48 h to obtain Klebsiella lemongrass liquid culture medium. The liquid seed culture medium contained the following components: 8 g / L beef extract, 5 g / L yeast powder, 10 g / L glucose, and natural pH. B2: Inoculate the liquid culture medium with 1%-3% of the inoculum, and ferment at 26℃-30℃, 200-220 r / min for 60-72 h. After filtration, obtain the fermentation broth and dilute it to 2×10⁻⁶. 7 CFU / ml was used to obtain lemon-shaped Klerk yeast inoculum; the composition of the liquid fermentation medium was the same as that of the liquid seed medium in step B1.

4. The method for controlling tomato gray mold according to claim 1, characterized in that: The compound microbial agent contains deep red yeast ( Rhodotorula Rubra ) inoculants and lemon-shaped Klerk yeast ( Kloeckera Apiculata The volume ratio of the bacterial agent is 1:

4.

5. The method for controlling tomato gray mold according to claim 1, characterized in that: The compound microbial agent contains deep red yeast ( Rhodotorula Rubra ) inoculants and lemon-shaped Klerk yeast ( Kloeckera Apiculata The volume ratio of the bacterial agent is 1:

3.

6. The method for controlling tomato gray mold according to claim 1, characterized in that: The compound microbial agent contains deep red yeast ( Rhodotorula Rubra ) inoculants and lemon-shaped Klerk yeast ( Kloeckera Apiculata The volume ratio of the bacterial agent is 1:6.