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Baclillus subtilis used for controlling cotton verticillium wilt and application thereof

A technology for cotton Verticillium wilt and Bacillus subtilis, applied in the field of Bacillus, can solve problems such as difficulty in control, difficulty in disease control methods and agents to achieve ideal control effects, and difficulty in finding resistant germplasm resources.

Inactive Publication Date: 2012-05-09
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 disease control methods and pesticides researched and developed 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 in 70% alcohol for 1 min, and sterilize the surface of the sample with 1.05% sodium hypochlorite for 15 minutes. ~18min, take 1.00g of the material that has been thoroughly sterilized on the surface of the test, put it in a sterile mortar, add sterilized PBS buffer solution to grind, let it stand for 15min, add buffer solution to 10ml, take 1ml of the separation solution prepared above and shake Shake vigorously on the bed for 30 minutes and then keep in a water bath at 80°C for 15-20 minutes. Then use sterile water to carry out gradient dilution, apply LB plates, and incubate in a 37°C incubator for about 48 hours. Pick single colonies with obvious differences in colony morphology, and after purification, store them on a slant to enter the preliminary screening procedure...

Embodiment 2

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

[0034] 2.1 Preparation of Bacillus subtilis fermentation broth

[0035] Extract 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, shake and culture at 28°C for 48 hours, centrifuge the above-mentioned Bacillus subtilis bacterial liquid (10000r / min, 20min), take the supernatant and filter it with a 0.22μm filter membrane to obtain the sterile fermentation liquid of the bacteria, Store in the refrigerator after sealing.

[0036] Inhibitory effect of 2.241B-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 liquid prepared in 2.1, and mix it evenly with the PDA medium cooled at about 45°C in different proportions. A...

Embodiment 3

[0042] Determination of the preliminary properties of antagonistic substances in the fermented liquid of embodiment 3

[0043] 3.1 Thermal stability

[0044] Take the 41B-1 (CGMCC No.5582) sterile fermentation broth prepared in 2.1, and put it in a water bath at 40°C, 60°C, 80°C, and 100°C for 30 minutes, and take the fermentation broth without water bath treatment as a control, and use plate diffusion The antibacterial activity against Verticillium dahliae Hn-3 was detected by the method, 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. Taking the untreated 41B-1 (CGMCC No.5582) aseptic fermentation broth as a control, the plate diffusion...

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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 Verficillium dahliae is the main disease of cotton, which greatly affects the yield and quality of cotton. 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 e...

Claims

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Application Information

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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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