Myxobacteria H56D21 preying on phytopathogen and application of myxobacteria H56D21

A technology of H56D21 and plant pathogenic bacteria, applied to the myxobacterium H56D21 that preys on plant pathogenic bacteria and its application fields, can solve many problems and achieve the effect of broad-spectrum predation characteristics

Active Publication Date: 2021-10-29
GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there are not many reports on the application of myxobacteria in the biological control of plant pathogens, and they mainly focus on myxococcus ( Myxococcus ), Pseudomonas ( Corallococcus ), polycystic bacteria ( Polyangium ), Cystobacillus ( Cystobacter ) and other genera, currently only 3 related patents can be retrieved in the patent database: a strain of myxobacteria and its applic...

Method used

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  • Myxobacteria H56D21 preying on phytopathogen and application of myxobacteria H56D21
  • Myxobacteria H56D21 preying on phytopathogen and application of myxobacteria H56D21
  • Myxobacteria H56D21 preying on phytopathogen and application of myxobacteria H56D21

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Isolation and purification of myxobacteria H56D21

[0028] 1.1 Isolation of strain H56D21

[0029] Medium: (1) ST21 Medium (Solution A: K 2 HPO 4 1.0 g, Yeast extract 0.02 g, Agar14.0 g, Water 600 mL; solution B: KNO 3 1.0 g, MgSO 4 ·7H 2 O 1.0 g, CaCl 2 2H 2 O 1.0 g, MnSO 4 ·7H 2 O 0.1 g, Water 300 mL; Solution C: FeCl 30.2 g, Water 100 mL. Solutions A, B, and C are sterilized separately, heated to about 50°C and mixed, then poured into the culture medium). (2) VY / 2 medium: angel yeast 5 g, CaCl 2 .2H 2 2。 O 1.36g, Agar 15 g, water 1L, mix the ingredients, adjust the pH 7.2. Add VB after sterilization 12 50 μg / mL. (3) LB medium: tryptone 10 g, NaCl 10 g, yeast powder 5 g, pH 7.2.

[0030] Weigh 10 g of air-dried forest soil samples, add an appropriate amount of sterile water (add cycloheximide with a final concentration of 100 µg / mL), soak for about 10 h, and place the soil samples on ST21 covered with sterile filter paper. After cultured ...

Embodiment 2

[0033] Example 2 Classification and Identification of Myxobacteria H56D21

[0034] 2.1 Morphological identification

[0035] See figure 1 . On ST21 medium, the fruiting body is solitary, pink, ellipsoidal or pea-shaped, and covered with rich mucus (a); on VY / 2 medium, the colony is pink or yellow-brown, round, and radiating Film-like, with flame-like protrusions on the edge (b); vegetative cells are Gram-negative rod-shaped cells with blunt ends (c).

[0036] 2.2 Molecular identification

[0037] The genomic DNA of strain H56D21 was extracted by the improved CTAB method, and the 16S rRNA gene was amplified by the bacterial universal primer F27 / 1492R. The amplified product was sent to Shanghai Meiji Biomedical Technology Co., Ltd. (Guangzhou Branch) for sequencing analysis. The length of the 16S rRNA gene sequence obtained by splicing with SeqMan software was 1536 bp, as shown in SEQ ID NO:1. The sequence was compared and analyzed in NCBI and Ezbiocloud (http: / / www.ezbioc...

Embodiment 3

[0045] Example 3 Myxobacterium H56D21 preys on plant pathogenic bacteria

[0046] Strain H56D21 was inoculated on VY / 2 solid medium and cultured at 30 °C for 7 days. The phytopathogenic bacterium R. solanacearum ( Ralstonia solanacearum ) GDMCC 1.1561, cabbage black rot ( Xanthomonas campestris ) GDMCC 1.857, Sweet pea pathogens ( Rhodococcus fascians ) GDMCC 1.839 were respectively transferred to liquid nutrient broth medium, 30 ℃, 180 r / min shaking culture for 48 h, then centrifuged at 8000 r / min for 5 min, the bacteria were collected, washed twice with MMC buffer, and the bacteria The body weight was suspended in MMC buffer, and the cell concentration was diluted to 1×10 11 cfu / mL, each take 150 µL of bacterial solution and spot in TPM medium (Tris-HCl (pH 7.6) 10 mM, K 2 HPO 4 1 mM, MgSO 4 8 mM, Agar 15g, Water 1 L) in the center to make the plaque round. After the bacteria were dry, holes were punched on the myxobacteria H56D21 colony with a blue pipette tip, ...

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Abstract

The invention discloses myxobacteria H56D21 preying on phytopathogen and application of the myxobacteria H56D21, and belongs to the field of microbial technology and biological control. The ANI value and the dDDH value of the strain H56D21 and the similar model strain are analyzed, and the result shows that the strain H56D21 is a new species of Hyalangium. The Hylalangium sp.H56D21 is preserved in the Guangdong Microbial Culture Collection Center on November 17, 2020, the address is the 5th floor of the building 59, No.100 courtyard, Xianlie Middle Road, Guangzhou City, Guangdong Province, the postcode is 510070, the preservation number is GDMCC No: 61294, and the Hylalangium sp.H56D21 can prey plant pathogenic bacteria, has broad-spectrum predation characteristics and can be used for biological control of plant diseases.

Description

technical field [0001] The invention relates to the field of microbial technology and biological control, in particular to a myxobacterium H56D21 that preys on plant pathogenic bacteria and its application. Background technique [0002] Plant diseases are one of the important factors that restrict the high-quality and high-yield of crops. While causing serious losses to crop yields, they also increase the input costs of agricultural production and restrict the improvement of the quality of agricultural products. At present, the main ways to prevent and control plant diseases include chemical pesticide control, breeding of resistant varieties, biological control and other means. The chemical pesticides widely used at present have quick effects, but they are easy to induce drug resistance of pathogenic bacteria and cause secondary pollution to the environment; the breeding of resistant varieties often takes a long period and is technically difficult; while biological control h...

Claims

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Application Information

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IPC IPC(8): A01N63/20A01P3/00A23L33/135A23K10/18C12N1/20C12R1/01
CPCA01N63/20C12N1/20A23L33/135A23K10/18A23V2002/00A23V2200/30
Inventor 朱红惠张鲜姣姚青冯广达
Owner GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY
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