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Acid-resistant pseudomonas protegens CLP-6 and application thereof

A CLP-6, Pseudomonas technology, applied in the field of microorganisms, can solve problems such as poor acid resistance, and achieve the effects of wide acidity tolerance, strong stress resistance, and broad antibacterial spectrum

Active Publication Date: 2018-09-25
TOBACCO RES INST CHIN AGRI SCI ACAD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although there have been many studies on the biological control of tobacco bacterial wilt and other fungal or bacterial diseases at home and abroad, and some results have been achieved, these strains are poor in acid resistance, and they will be hardened and acidified when they are applied to acidic soil, especially if they are continuously cropped. In severe soils, its antagonistic effect cannot be fully exerted, and most biocontrol strains only have good control effects on one type of fungal or bacterial diseases, while the defense false ones are acid-resistant and have biocontrol effects on soil-borne disease pathogens. Single cell strains have not been reported yet

Method used

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  • Acid-resistant pseudomonas protegens CLP-6 and application thereof
  • Acid-resistant pseudomonas protegens CLP-6 and application thereof
  • Acid-resistant pseudomonas protegens CLP-6 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Example 1 Verification of the antagonistic activity of CLP-6 and its fermented liquid against tobacco bacterial wilt and black shank pathogens: the antagonistic activity of the diseased tobacco strains from Jiangya Town, Cili County, Zhangjiajie City, Hunan Province was verified. Mix the NA medium with an appropriate amount of acidic buffer solution (0.2mol / L sodium acetate and 0.3mol / L acetic acid mixed at a volume ratio of 1:9) to prepare different buffers with pHs of 5.5, 6.0, 6.5, 7.0 and 7.5, respectively. pH NA medium, using the Oxford cup method to determine the difference in antibacterial activity of the fermentation broth of CLP-6 strain under different pH conditions. Inoculate CLP-6 on the NA plate and culture it at 30°C for 48 hours, and then use the plate confrontation method, that is, inoculate the blackleg bacteria cake (5 mm in diameter) and Alternaria rubella on the oat and potato dextrose medium (PDA) plate, and use the inoculation loop Pick CLP-6 and d...

Embodiment 2

[0073] Example 2 CLP-6 and its fermented liquid damage the bacterial body of Ralstonia solanacearum: Inoculate CLP-6 on NA plate and cultivate at 30°C for 48 hours, then use the plate confrontation method, and use Oxford cup plate for the inhibition of Ralstia solanacearum Diffusion method: inoculate CLP-6 in beef juice peptone liquid culture medium (NB), shake culture at 30°C and 150rpm for 2 days, take appropriate amount of fermentation broth and sterile supernatant (filtered through a bacterial filter) In the Oxford cup on the beef juice peptone medium plate (NA) of the dead bacteria, it was cultured statically for 2 days, and the inhibition of CLP-6 to the above-mentioned pathogenic bacteria was observed. Observing from the indoor plate antibacterial effect, CLP-6 thalline, fermented liquid and aseptic supernatant have good inhibitory effect to R. solanacearum of tobacco (see figure 2 ).

[0074] The antibacterial activity of the bacteriostatic substances secreted by the...

Embodiment 3

[0075] Example 3 CLP-6 and its fermented liquid inhibit the mycelial growth of black shank fungus: take black shank fungus as the target bacteria, inoculate CLP-6 on the NA plate for 48 hours at 30 ° C, and then use the plate confrontation method, that is, in the Inoculate black shank bacteria on the oat plate, pick CLP-6 with an inoculation loop and draw equidistant lines with the fungus cake as the center, culture at 30°C for 5 days, pick the hyphae from the edge of the black shank fungus cake and examine under a microscope, The result shows that compared with normal hyphae, the cytoplasm in the hyphae cells of the black shank bacteria cake edge hyphae after CLP-6 processing is aggregated into a spherical shape, which breaks away from the cell membrane and cell wall, thus affecting its normal growth (see Figure 4 ).

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Abstract

The invention provides acid-resistant pseudomonas protegens CLP-6 and application thereof. According to the acid-resistant pseudomonas protegens CLP-6 and the application thereof, the bacterial strainof the pseudomonas protegens CLP-6 is preserved in China General Microbiological Culture Collection Center on October 27th, 2016, and the biological preservation number of the pseudomonas protegens CLP-6 is CGMCC No.13205; the bacterial strain of the pseudomonas protegens CLP-6 has acid resistance, is good in disease prevention and plant growth promotion effects and high in stress resistance, hasthe antagonistic activity of pathogenic bacteria, is wide in soil acidity tolerance range and wide in antibacterial spectrum, and has application and development potential in prevention and treatmentof bacterial wilt, black shanks and brown spots; and meanwhile, under the general background that the acidification conditions in China is increasingly severe, the research on the bacterial strain ofthe pseudomonas protegens CLP-6 has reference significance in prevention and treatment of plant diseases under various soil qualities, a microbial resource is provided for green prevention and control over the bacterial wilt, the black shanks and the brown spots in acid soil, and secondary metabolites of microorganisms are excavated, so that an important foundation can be laid for the biocontrolefficacy on the bacterial wilt, the black shanks and the brown spots in the acid soil.

Description

technical field [0001] The invention belongs to the technical field of microbes, and in particular relates to an acid-resistant strain of defensive pseudomonas CLP-6 and its application. Background technique [0002] Tobacco will suffer from a variety of fungal and bacterial diseases throughout the growth period. There are more than 60 kinds of tobacco diseases recorded in China, of which nearly 10 are the most serious, mainly including tobacco black shank, bacterial wilt, red spot, and anthracnose , black disease and powdery mildew. Some pathogens can damage tobacco at various growth stages. Rhizome diseases such as tobacco black shank (Phytophthoraparasitica var. Nicotianae) and tobacco bacterial wilt (Ralstonia solanacearum) can damage tobacco from seedling to maturity. In the later period, the damage was serious, and it became popular with the climate of different years, resulting in a serious reduction in the production of tobacco fields and seriously affecting the yie...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/00A01P1/00A01P3/00A01P21/00C05F11/08C05G3/00C12R1/38
CPCA01N63/00C05F11/08C05G3/00C12N1/20C12N1/205C12R2001/38
Inventor 王静彭德元李斌王振华王贻鸿赵倩曾庆宾黎妍妍夏博
Owner TOBACCO RES INST CHIN AGRI SCI ACAD
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