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Bacillus substilis BS01 and inoculant thereof, as well as application of bacillus substilis BS01 to inhibition of picked fruit pathogenic bacteria

A technology of Bacillus subtilis and fruit, applied in the field of microbiology, can solve the problems of unstable control effect, long storage time, and reduced activity, and achieve the effects of avoiding long storage time, reduced activity, simple preparation method, and low industrialization cost

Inactive Publication Date: 2017-02-22
POMOLOGY RES INST GUANGDONG ACADEMY OF AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The use of live Bacillus subtilis has problems such as long storage time, decreased activity, unstable adaptability to the environment, difficulty in colonization, unstable control effect, large effective dose, and narrow antibacterial spectrum.

Method used

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  • Bacillus substilis BS01 and inoculant thereof, as well as application of bacillus substilis BS01 to inhibition of picked fruit pathogenic bacteria
  • Bacillus substilis BS01 and inoculant thereof, as well as application of bacillus substilis BS01 to inhibition of picked fruit pathogenic bacteria
  • Bacillus substilis BS01 and inoculant thereof, as well as application of bacillus substilis BS01 to inhibition of picked fruit pathogenic bacteria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Isolation and Identification of Bacillus subtilis BS01:

[0055] Bacillus subtilis (Bacillus substilis) BS01 was isolated from the soil of the Banana Experimental Garden in Dongguan, Guangdong, and was separated according to the principle of high temperature resistance of its spores. The method is: take about 100g of soil with a surface layer of 5-10cm, put it into a paper sample bag Return to the laboratory for separation; weigh 5g of soil and put it into a triangular flask containing 50mL of water, shake it for a while, then slowly heat it to boiling, boil for 15min and then let it stand for 10min, take 100μL of the upper layer solution and use sterile water for gradient dilution, and then separately draw 10 -2 and 10 -3 Spread 200 μL of the diluted solution on the beef broth-peptone medium, culture at 37°C for 2 days, and then purify; pick some bacteria from a single colony on the plate, and carry out multiple streak cultures on the same medium, and after the colony...

Embodiment 2

[0058] The fermentation culture method of bacillus subtilis BS01 comprises the following steps:

[0059] (1) Activation of preserved strains: Bacillus subtilis BS01 strains were stored in an ultra-low temperature refrigerator, and the strains were taken out and thawed on ice, and then inoculated with an inoculation needle in LB (containing 10 g of tryptone per liter, 5 g of yeast extract powder, NaCl 10g) Stretch culture on the flat plate, BS01 single colony grows on the LB after 1 day;

[0060](2) Preparation of seed liquid: Pick a single colony from the above-mentioned LB solid plate to 50 mL of LB liquid medium, 37 ° C, 200 rpm, and cultivate for 24 hours to obtain BS01 fermented seed liquid;

[0061] (3) Fermentation culture: adjust the concentration of the seed liquid prepared above with sterile water to 6.5×10 7 cfu / mL, transferred to the optimized fermentation medium at a ratio of 10%; the liquid volume of the shake flask was 1 / 5 of the bottle volume, and then 30°C, 18...

Embodiment 3

[0065] Compared with Example 2, it is the same as Example 2 except that the optimized fermentation medium is replaced with the following composition.

[0066] The optimized fermentation medium is composed of the following components: glucose 3.0%, bacto-peptone 2.0%, sodium chloride 0.5%.

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Abstract

The invention relates to bacillus substilis BS01 and an inoculant thereof, as well as application of the bacillus substilis BS01 to inhibition of picked fruit pathogenic bacteria. The bacillus substilis BS01 is preserved in Microbial Culture Preservation Center, Guangdong Province (GDMCC) on August, 29, 2016, and the preservation number is GDMCC NO.60069. The in-vitro antibacterial activity of the supernatant of the fermenting liquid of the bacillus substilis BS01 strain provided by the invention on the picked fruit pathogenic bacteria, particularly colletotrichum, can reach 90 percent or above, and is about three times of the antibacterial activity of the supernatant of the fermenting liquid generated by fermentation of the common LB culture medium before optimization; and the inhibition effect of the fermenting liquid of the BS01 strain obtained by the fermentation condition after optimization on anthracnose, such as banana anthracnose, tomato anthracnose and avocado anthracnose, can reach 65 percent or above.

Description

technical field [0001] The invention relates to the field of microbiology, in particular to a Bacillus subtilis (Bacillus substilis) BS01, its bacterial agent and its application in inhibiting postharvest pathogenic bacteria of fruit. Background technique [0002] Anthracnose (Colletotrichum spp.) is one of the main pathogens causing postharvest diseases of most fruits and vegetables. It occurs very commonly in fruits such as mango, papaya, avocado, guava, grass mold, etc. Postharvest losses of these fruits Most of this is due to disease and rot caused by postharvest pathogenic fungi, including anthracnose. At present, the control of postharvest pathogenic fungi mainly uses chemical fungicides targeting sterol synthesis. However, long-term and high-frequency use of a class of fungicides often leads to drug resistance of pathogenic bacteria, which has led to a decline in the control effect, or even failure. Moreover, the application of chemical fungicides has problems such ...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/00A01N63/02A01P3/00C12R1/125
CPCA01N63/00A01N63/10C12N1/20C12N1/205C12R2001/125
Inventor 孟祥春毕方铖章斌黄泽鹏
Owner POMOLOGY RES INST GUANGDONG ACADEMY OF AGRI SCI
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