Pseudomonas choloeaphtis and application

Abstract

The invention relates to pseudomonas choloeaphtis with a preservation number of CGMCC: 8431 and application thereof. Pseudomonas choloeaphtis can be further made into a biocontrol microbial inoculum for controlling wheat take-all disease with good control effect, and helps to make up the disadvantage of bad control effect of chemical agents. Because pseudomonas choloeaphtis strains can be colonized at the wheat rhizosphere, the inoculant has a relatively long lasting period, and the inoculant does not bring environmental problems and is beneficial to pollution-free production of wheat.

Description

technical field

[0001] The invention relates to the field of microorganisms, in particular to Pseudomonas chloropinus and its application. Background technique

[0002] Wheat take-all (Take-all), also known as "black foot" disease, is a typical root soil-borne disease, mainly caused by wheat variety Gaeumannomyces graminis var. tritici (Ggt). Pathogens generally infect wheat roots and 1-2 nodes at the base of stems. Wheat take-all can occur at both seedling and mature stages. Symptoms are more obvious at the jointing stage, with leaves turning yellow and roots turning black. Period, white spike symptoms appear.

[0003] Wheat take-all was first discovered in South Australia in 1852. Later, wheat take-all was found on the dead stems of bermudagrass and wheatgrass in northern Italy. After that, it appeared in various countries and became more and more serious, causing huge losses to wheat production. . In 1931, wheat take-all was discovered for the first time in Zhejiang...

Examples

Embodiment 1

[0015] Example 1 Isolation and screening of Pseudomonas chloropinus bacterial strains

[0016] 1. Isolation of Pseudomonas chloropinus

[0017] Take 10g of the soil sample adsorbed on the wheat root system, add 90ml of sterile water, shake fully, and get 10 -1 Diluent; continue to dilute with sterile water to obtain 10 -5 、10 -6 and 10 -7 Take 0.2ml of the above three dilutions and spread them on the LB plate, incubate at 25°C for 48 hours, pick a single colony and inoculate it on the slant of LB solid medium for preservation;

[0018] 2. Screening of Pseudomonas chloropinus

[0019] A) Inoculate the bacteria on each slant in step 1 into 1ml LB liquid medium, and shake and culture at 28°C 120r / min for 12 hours;

[0020] B) Inoculate the 5mm dish of the highly pathogenic strain G1037 (hereinafter referred to as G1037) of Gaeumannomyces graminis var. tritici to the center of the PDA plate, and after 48 hours of cultivation, the four sides of the four sides of the G1037 c...

Embodiment 2

[0023] Physiological and biochemical analysis of embodiment 2 GP51 bacterial strain

[0024]

[0025] Note: + is positive, - is negative

Embodiment 3

[0026] Example 3 Molecular biological detection of antibiotic synthesis-related genes in the genome of GP51 strain

[0027] The genes related to the synthesis of four antibiotics in the genome of GP51 strain were detected by PCR method: 2,4-diacetylphloroglucinol (phlD), phenazine (phzCD), pyocyanin (pltC), nitric acid Pyrrolactin (prnC). These four antibiotics are the main classes of antibiotics produced by Pseudomonas bacteria. The primers used are as follows:

[0028]

[0029] The genes related to the synthesis of phenazine and nipyrrolidin could be detected in the genome of GP51 strain; the genes related to the synthesis of 2,4-diacetylphloroglucinol and pyocyanin were not detected. In order to further confirm whether the detected genes are target genes, the detected genes were cloned and sequenced respectively, and the obtained sequences were subjected to BLAST on the NCBI website.

[0030] The sequence obtained with phzCD primers and the accession number is HM59428...

technical field

[0001] The invention relates to the field of microorganisms, in particular to Pseudomonas chloropinus and its application. Background technique

[0002] Wheat take-all (Take-all), also known as "black foot" disease, is a typical root soil-borne disease, mainly caused by wheat variety Gaeumannomyces graminis var. tritici (Ggt). Pathogens generally infect wheat roots and 1-2 nodes at the base of stems. Wheat take-all can occur at both seedling and mature stages. Symptoms are more obvious at the jointing stage, with leaves turning yellow and roots turning black. Period, white spike symptoms appear.

[0003] Wheat take-all was first discovered in South Australia in 1852. Later, wheat take-all was found on the dead stems of bermudagrass and wheatgrass in northern Italy. After that, it appeared in various countries and became more and more serious, causing huge losses to wheat production. . In 1931, wheat take-all was discovered for the first time in Zhejiang...

Examples

Embodiment 1

[0015] Example 1 Isolation and screening of Pseudomonas chloropinus bacterial strains

[0016] 1. Isolation of Pseudomonas chloropinus

[0017] Take 10g of the soil sample adsorbed on the wheat root system, add 90ml of sterile water, shake fully, and get 10 -1 Diluent; continue to dilute with sterile water to obtain 10 -5 、10 -6 and 10 -7 Take 0.2ml of the above three dilutions and spread them on the LB plate, incubate at 25°C for 48 hours, pick a single colony and inoculate it on the slant of LB solid medium for preservation;

[0018] 2. Screening of Pseudomonas chloropinus

[0019] A) Inoculate the bacteria on each slant in step 1 into 1ml LB liquid medium, and shake and culture at 28°C 120r / min for 12 hours;

[0020] B) Inoculate the 5mm dish of the highly pathogenic strain G1037 (hereinafter referred to as G1037) of Gaeumannomyces graminis var. tritici to the center of the PDA plate, and after 48 hours of cultivation, the four sides of the four sides of the G1037 c...

Embodiment 2

[0023] Physiological and biochemical analysis of embodiment 2 GP51 bacterial strain

[0024]

[0025] Note: + is positive, - is negative

Embodiment 3

[0026] Example 3 Molecular biological detection of antibiotic synthesis-related genes in the genome of GP51 strain

[0027] The genes related to the synthesis of four antibiotics in the genome of GP51 strain were detected by PCR method: 2,4-diacetylphloroglucinol (phlD), phenazine (phzCD), pyocyanin (pltC), nitric acid Pyrrolactin (prnC). These four antibiotics are the main classes of antibiotics produced by Pseudomonas bacteria. The primers used are as follows:

[0028]

[0029] The genes related to the synthesis of phenazine and nipyrrolidin could be detected in the genome of GP51 strain; the genes related to the synthesis of 2,4-diacetylphloroglucinol and pyocyanin were not detected. In order to further confirm whether the detected genes are target genes, the detected genes were cloned and sequenced respectively, and the obtained sequences were subjected to BLAST on the NCBI website.

[0030] The sequence obtained with phzCD primers and the accession number is HM59428...

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