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Baclillus subtilis used for controlling cotton verticillium wilt and application thereof

A technology of cotton Verticillium wilt and Bacillus subtilis, which is applied in the field of Bacillus, can solve the problems of difficulty in finding germplasm resources of resistance, disease control methods and agents that are difficult to achieve ideal control effects, and control difficulties

Inactive Publication Date: 2013-04-10
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For the prevention and treatment of cotton Verticillium wilt, a lot of work has been done in disease-resistant breeding, agricultural measures and chemical control at home and abroad. Cotton Verticillium wilt is a soil-borne vascular disease, and it is difficult to control and find good resistant species. As a result, new cotton varieties with high resistance to Verticillium wilt have not been bred for many years, and the research and development of disease control methods and pesticides are also difficult to achieve the desired control effect

Method used

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  • Baclillus subtilis used for controlling cotton verticillium wilt and application thereof
  • Baclillus subtilis used for controlling cotton verticillium wilt and application thereof
  • Baclillus subtilis used for controlling cotton verticillium wilt and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] 1.1 Isolation of bacteria

[0026] Take the main root of the cotton disease-resistant strain R104 at the budding stage that grows healthy, take the main root at 5-10 cm with a sterilized blade, immerse it in 70% alcohol for 1 min, and sterilize the surface of the sample with 1.05% sodium hypochlorite. For 15-18 min, take 1.00 g of the material that has been thoroughly sterilized on the surface of the test, put it in a sterile mortar, add sterilized PBS buffer, grind it, let it stand for 15 min, add buffer to make it to 10 ml, take the prepared above Shake 1 ml of the separation solution vigorously on a shaker for 30 min, then place it in a water bath at 80°C for 15-20 min. Then, after gradient dilution with sterile water, LB plates were spread, and cultured in a 37°C incubator for about 48 hours. Single colonies with obvious differences in colony morphology were picked, purified and stored on slant surfaces to enter the preliminary screening procedure.

[0027] 1.2 Scr...

Embodiment 2

[0033] Example 2 Study on the Antagonism Mechanism of Bacillus phleiformis 41B-1 (CGMCC No.5582)

[0034] 2.1 Preparation of Bacillus subtilis fermentation broth

[0035] Extract a single colony of 41B-1 (CGMCC No.5582) with a toothpick to liquid LB medium (peptone 10g, yeast extract 5g, Nacl 10g, distilled water 1000ml, pH 7.0), 37°C, 12h, and then inoculate with 1% inoculum Transfer to fresh LB medium, culture with shaking at 28°C for 48 hours, centrifuge the above-mentioned Bacillus subtilis bacterial liquid (10000 r / min, 20 min), take the supernatant and filter it with a 0.22 μm filter membrane to obtain the sterile fermentation liquid of the bacteria , sealed and placed in the refrigerator for later use.

[0036] 2.2 Inhibitory effect of 41B-1 (CGMCC No.5582) fermentation broth on the mycelia growth of Verticillium dahliae Hn-3

[0037] Take the 41B-1 (CGMCC No.5582) sterile fermentation broth prepared in 2.1, and mix it evenly with the PDA medium cooled at about 45°C i...

Embodiment 3

[0042] Example 3 Determination of preliminary properties of antagonistic substances in fermentation broth

[0043] 3.1 Thermal stability

[0044] Take the 41B-1 (CGMCC No.5582) sterile fermentation broth prepared in 2.1, and put them in water baths at 40°C, 60°C, 80°C, and 100°C for 30 min, respectively, and use the fermentation broth without water bath treatment as a control, use a plate Diffusion method was used to detect its antibacterial activity against cotton Verticillium dahliae Hn-3, and the results showed that the antagonistic substances had strong thermal stability (Table 2).

[0045] Table 2. Effects of different temperatures on the antibacterial activity of antagonistic substances

[0046]

[0047] 3.2 Stability to pH

[0048] Take the 41B-1 (CGMCC No.5582) sterile fermentation broth prepared in 2.1, and adjust its pH to 5.0, 7.0, 10.0 and 12.0, respectively. Using the untreated 41B-1 (CGMCC No.5582) aseptic fermentation broth as a control, the plate diffusio...

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Abstract

The invention relates to Baclillus subtilis used for controlling cotton verticillium wilt and application thereof, belonging to the technical field of biological pesticides. Baclillus subtilis 41B-1 is preserved in China General Microbiological Culture Collection Center on December 13, 2011 and is assigned the accession number of CGMCC No. 5582. The invention relates to application of Baclillus subtilis 41B-1 with an accession number of CGMCC No. 5582 in controlling cotton verticillium wilt. Baclillus subtilis 41B-1 (CGMCC No. 5582) is non-toxic to human beings and animals, nonhazardous to crops, green and environment-friendly and has a remarkable effect on controlling cotton verticillium wilt and a good prospect in development of biological pesticides.

Description

technical field [0001] The invention belongs to the technical field of biological pesticides, and relates to a bacillus strain used for preventing and treating Verticillium wilt of cotton and its application. Background technique [0002] Verticillium dahliae caused by Verticillium dahliae is the main disease of cotton, which has a great impact on cotton yield and quality. For the prevention and treatment of cotton Verticillium wilt, a lot of work has been done in disease-resistant breeding, agricultural measures and chemical control at home and abroad. Cotton Verticillium wilt is a soil-borne vascular disease, and it is difficult to control and find good resistant species. As a result, new cotton varieties with high resistance to Verticillium wilt have not been bred for many years, and the disease control methods and pesticides researched and developed are also difficult to achieve the desired control effect. At present, biological control is considered to be one of the ef...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/02A01P3/00C12R1/125
Inventor 唐灿明赵明文韩琴李顺鹏
Owner NANJING AGRICULTURAL UNIVERSITY
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