A Myxobacterium h56d21 Predating Phytopathogenic Bacteria and Its Application
A technology of H56D21 and plant pathogenic bacteria, which is applied to myxobacteria H56D21 that prey on plant pathogenic bacteria and its application field, can solve many problems and achieve the effect of broad-spectrum predation characteristics
Image
Examples
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, ...
PUM
Login to View More Abstract
Description
Claims
Application Information
- IPC
- A01N63/20; A01P3/00; A23L33/135; A23K10/18; C12N1/20; C12R1/01
- CPC
- A01N63/20; C12N1/20; A23L33/135; A23K10/18; A23V2002/00; A23V2200/30
- Inventors
- 朱红惠; 张鲜姣