Bacillus pumilus as well as cultural method and application thereof

A technology of Bacillus pumilus and its cultivation method, which is applied in the field of biological strains, achieves the effect of significant curative effect and obvious antagonistic effect

Active Publication Date: 2014-04-16
SHANDONG PEANUT RES INST +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

my country's research on peanut antibacterial agent is relatively late, but it develops rapidly

Method used

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  • Bacillus pumilus as well as cultural method and application thereof
  • Bacillus pumilus as well as cultural method and application thereof
  • Bacillus pumilus as well as cultural method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1 Bacillus pumilus LX11 strain culture and separation

[0026] The formula of LB solid medium was: tryptone 10g / L, yeast extract 5g / L, sodium chloride 10g / L and agar powder 15g / L, sterilized at 121°C for 15min before use.

[0027] Cultivation method: Weigh 5g of peanut rhizosphere soil into a triangular flask with 45ml of distilled water, shake it for a while, absorb 0.1ml of the supernatant and add it to a test tube containing 4.5ml of sterile water to make a suspension with a concentration of 1:100. draw 10 -2 and 10 -3 Add 0.2ml of the soil suspension solution dropwise to the LB solid medium plate containing rifampicin, spread it evenly with a spreading rod, and place the medium upside down in a constant temperature incubator at 28°C for 1-2 days.

[0028] Isolation of bacterial strains: Pick a batch of single colonies on the plate, draw a line on the LB solid medium plate and place it upside down in a constant temperature incubator at 28°C for 1 to 2 da...

Embodiment 2

[0029] Identification of embodiment 2LX11 bacterial strain

[0030] (1) Strain DNA extraction:

[0031] Refer to the TIANGEN TIANampBACTERia DNA Kit kit to extract the total DNA of the LX11 strain in the examples, the steps are as follows:

[0032] 1. Take 1mL of bacterial culture solution, centrifuge at 10,000rpm for 1min, and absorb the supernatant as much as possible;

[0033] 2. Add 200 μL buffer GA to the cell pellet and shake until the cell is completely suspended;

[0034]3. Add 4μL RNAase (100mg / mL) solution, shake for 15s, and place at room temperature for 5min;

[0035] 4. Add 20 μL proteinase K solution to the tube and mix well;

[0036] 5. Add 220 μL buffer GB, shake for 15 seconds, place at 70°C for 10 minutes, and briefly centrifuge to remove water droplets on the inner wall of the tube cap;

[0037] 6. Add 220 μL of absolute ethanol, shake and mix well for 15 seconds. At this time, flocculent precipitation may appear, and briefly centrifuge to remove water d...

Embodiment 3

[0050] Embodiment 3LX11 bacterial strain suppresses fungus ability

[0051] 3.1 Culture of fungi

[0052] Use tweezers to inoculate peanut root rot fungus, peanut rot fungus and peanut stem rot fungus into PDA medium with tweezers, and inoculate them upside down in a greenhouse at 28°C for 3 days.

[0053] 3.2 Bacteria inoculation

[0054] When the fungus grows to about 1 / 3 of the size of the petri dish, inoculate the strain of Example 2 at a distance of 1-2 cm from the fungus, and continue to culture in a greenhouse at 28 degrees Celsius for 2-3 days to observe the growth of the fungus.

[0055] Inhibition rate = [(control fungal growth radius - treatment fungal growth radius) / control fungal growth radius] × 100%.

[0056] The confrontation test of LX11 strain on peanut root rot, white silkworm and stem rot was carried out three times, and the inhibitory effect was obvious (see Table 1, 2, 3), which shows that LX11 has obvious antagonistic effect on pathogenic bacteria, and...

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Abstract

The invention discloses a bacillus pumilus LX11 strain as well as a cultural method and application thereof. The bacillus pumilus LX11 strain disclosed by the invention has a remarkable effect on preventing and controlling peanut root rot, southern blight and stem rot, an obvious antagonistic effect on pathogenic bacteria and a remarkable effect on preventing and controlling peanut diseases in pots and in the fields.

Description

technical field [0001] The invention relates to the technical field of biological strains, in particular to a bacillus pumilus, a cultivation method and an application of the bacillus pumilus. Background technique [0002] Peanut root rot is one of the most harmful and most difficult soil-borne diseases that occurs worldwide. In recent years, the disease has shown a trend of expanding and spreading in the main peanut producing areas of the world, and it has also aggravated year after year in the peanut producing areas in northern my country. In 2006, the disease occurred on a large scale in Xingtang County, Xinle City, Daming County and Dingzhou City of Hebei Province, Puyang City of Henan Province, Qingdao City, Rizhao City, Linyi City and Tai'an City of Shandong Province, and the incidence rate of severe disease fields exceeded 50%; In 2009, the epidemic in the northern peanut production area caused a significant reduction in peanut production, and some plots in Junan Cou...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/00A01P3/00C12R1/07
Inventor 迟玉成许曼琳黄志明孙春龙王丽宁王学武袁宗英刘爱娜吴菊香许婷婷谢宏峰陈殿绪焦坤袁美
Owner SHANDONG PEANUT RES INST
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