Bacillus velezensis as well as culture method and application thereof

A technology of Bacillus velesi and Bacillus, applied in the field of microorganisms, can solve problems such as sweet cherries that have not yet been consulted, and achieve the effects of improving fruit stress resistance, good inhibition effect, and delaying the process of spoilage

Active Publication Date: 2020-11-13
SHANDONG INST OF POMOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among the antagonistic microorganisms for postharvest preservation of sweet cherries, exogenous antagonistic strains such as Cryptococcus laurentii and Bacillus spp. have been reported in China, but no endogenous antagonistic strains of sweet cherries have been found so far. Strains reported

Method used

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  • Bacillus velezensis as well as culture method and application thereof
  • Bacillus velezensis as well as culture method and application thereof
  • Bacillus velezensis as well as culture method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Embodiment 1, isolation and identification of pathogenic bacteria and verification of pathogenicity

[0054] 1.1 Isolation and identification of pathogenic bacteria

[0055] Carry out surface disinfection with 75% medical alcohol of rotten sweet cherry sample, then wash the surface 3 times with sterile water, air-dry naturally, cut a small amount of rotten fruit tissue with sterile blade and tweezers, place it in resistant (streptomycin , penicillin) in the middle of the PDA medium, and cultivated at a constant temperature of 28°C for 3-4d. After the plate colony grows, pick the edge bacterial block and place it in another PDA medium, cultivate it at a constant temperature of 28°C, and purify it several times until a single colony is reached. A single colony was inoculated on the slant of the test tube and stored in a 4°C refrigerator for later use. All the above operations were carried out in a sterile environment.

[0056] The isolated and purified pathogenic bacte...

Embodiment 2

[0067] Embodiment 2, separation, screening and identification of antagonistic bacteria

[0068] 2.1 Isolation and screening of endogenous antagonistic bacteria

[0069] Take 30 healthy "Meizao" sweet cherry whole fruits and grind them, weigh 1g of the pulp and add it to 9mL sterile saline, mix well to make a stock solution, and let it stand for 20 minutes. Using serial dilution method, dilute to 10 -3 、10 -4 、10 -5 Take 100 μL of the dilution solution of the above concentration and spread it in LB medium, set 3 parallels for each gradient, and incubate at a constant temperature of 37°C for 24h. The grown colonies were inoculated onto another LB medium for three-section streaking, and incubated at a constant temperature of 37°C until a single colony grew. Pick a single colony to the LB slant, and store it at 4°C after the colony grows.

[0070] Using the filter paper method, take a 6mm bacterial cake on a PDA plate where the pathogenic bacteria Rhizopus stoloniferum was cu...

Embodiment 3

[0081] Embodiment 3, optimization of antagonistic bacteria culture medium

[0082] 3.1 The effect of carbon source on the growth of endogenous antagonistic bacteria

[0083] Pick the activated antagonistic bacteria Bacillus Veles Q-84, and cultivate the seed solution in LB medium. The NB medium was used as the basal medium, and the carbon source in the NB medium was replaced with 3 g / L sucrose, glucose, lactose, soluble starch and maltose, and the other culture components remained unchanged, and the NB medium was used as a control. Draw 1mL of the seed solution and inoculate it into 50mL of fermentation medium with different carbon sources, place it in a constant temperature shaker at 37°C, shake it at 180r / min for 16h, and measure the concentration of bacteria in the fermentation broth. The result is as Figure 6 As shown, when soluble starch was used as the only carbon source, it was most suitable for the growth of Bacillus velei Q-84, which was significantly different fro...

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Abstract

The invention relates to bacillus velezensis as well as a culture method and application thereof. The endophytic antagonistic strain bacillus velezensis TA-3-BV of sweet cherries is found for the first time, and the nucleotide sequence of 16S rDNA of the bacillus velezensis TA-3-BV is shown as SEQ ID NO.1. The bacillus velezensis TA-3-BV has broad-spectrum antibacterial activity on sweet cherries,has an inhibition effect on various pathogenic fungi and has an excellent inhibition effect on sweet cherry soft rot pathogenic bacteria rhizopus stolonifer, and the inhibition rate can reach 69.82 +/-1.43%. Besides, the bacillus velezensis TA-3-BV has a relatively good inhibition effect on postharvest decay of the sweet cherries, the decay process of the fruits can be delayed, the decay of the fruits is delayed by 2-3 days, and a novel method is provided for postharvest preservation and fresh keeping of the sweet cherries.

Description

technical field [0001] The invention relates to a strain of Bacillus Velez and its culture method and application, belonging to the technical field of microbes. Background technique [0002] Sweet cherry, also known as big cherry and cherry, is a plant of the Rosaceae (Rosaceae), Prunus, and Cherry subgenus (Cerasus). Sweet cherries are rich in nutritional value. The soluble protein content is about 589mg / 100g, and there are more Vc and various trace elements. It is rich in iron, about 8mg / 100g. It has the effect of anti-cancer, anti-tumor and promoting hemoglobin regeneration . Sweet cherry pulp is rich in a variety of phenolic compounds, including hydroxycinnamic acid, proanthocyanidins, flavone aldehydes and anthocyanins, which have strong antioxidant capacity and play a very important role in disease prevention and human health. [0003] Spoilage caused by fungal pathogenic microorganisms is a major problem in the production of cherry industry, and the loss caused by r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23B7/155A01N63/22A01P3/00C12R1/07
CPCC12N1/20A23B7/155A01N63/22A23V2002/00C12R2001/07C12N1/205A23V2200/10A01P3/00
Inventor 张倩辛力陈义伦吴澎郗良卿陈雨诗邹曼杨娟侠
Owner SHANDONG INST OF POMOLOGY
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