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An acid-resistant Pseudomonas defensensii clp-6 and its application

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: 2020-07-31
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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  • An acid-resistant Pseudomonas defensensii clp-6 and its application
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  • An acid-resistant Pseudomonas defensensii clp-6 and its application

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

[0074] Example 2 The damage degree of CLP-6 and its fermented liquid to Ralstia solanacearum: Inoculate CLP-6 on NA plate and culture at 30°C for 48 hours, then use plate confrontation method, use Oxford cup to inhibit Ralstia solanacearum Plate diffusion method: Inoculate CLP-6 in beef juice peptone liquid culture solution (NB), shake culture at 30°C and 150rpm for 2 days, take an appropriate amount of fermentation broth and sterile supernatant (filtered through a bacterial filter) The beef juice peptone medium plate (NA) of R. solanacearum was cultured statically in an Oxford cup for 2 days, and the inhibition of CLP-6 on 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 ).

[0075] The antibacterial activity of the bacteriostatic substances secreted by the CLP-6 strain on the destruction ...

Embodiment 3

[0076] Example 3 Inhibitory effect of CLP-6 and its fermented liquid on the mycelium growth of black shank fungus: take black shank fungus as target bacteria, inoculate CLP-6 on NA plate at 30°C for 48 hours, and then use the plate confrontation method, that is 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 edge hyphae of the black shank fungus cake and examine under a microscope , the results showed that compared with normal hyphae, the cytoplasm in the hyphae cells of the blackleg bacteria cake edge hyphae treated with CLP-6 was spherical and separated from the cell membrane and cell wall, thus affecting its normal growth (see Figure 4 ).

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Abstract

The invention provides a strain of acid-resistant Pseudomonas defensens CLP‑6 and its application. The strain was deposited in the General Microorganism Center of China Committee for Microorganism Culture Collection on October 27, 2016, and its biological preservation number is CGMCC No.13205. The bacterial strain has acid resistance, good disease prevention and growth-promoting effects, and strong stress resistance. The bacterial strain has pathogenic bacteria antagonistic activity, and has a wide range of tolerance to soil acidity and a wide antibacterial spectrum. It is effective in tobacco bacterial wilt. It has application and development potential in the prevention and treatment of black shank, black shank and red spot disease. At the same time, under the background of increasingly severe soil acidification in my country, the research on this strain is also of reference significance for the control of plant diseases in various soils, and it is a green control of tobacco bacterial wilt, black shank and red spot disease in acidic soils. It lays an important foundation for controlling and providing microbial resources, mining microbial secondary metabolites, and improving the biological control effects of bacterial wilt, black shank and red spot disease in acidic soils.

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 Patents(China)
IPC IPC(8): C12N1/20A01N63/27A01P1/00A01P3/00A01P21/00C05F11/08C05G3/60C12R1/38
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