Bacterial strain capable of antagonizing botrytis cinerea pathogens after picking of fruits and vegetables and application of bacterial strain

A technology for pathogenic bacteria and botrytis cinerea, which is applied in the field of microorganisms to achieve the effect of good inhibition and strong inhibition.

Active Publication Date: 2020-01-17
FARM PROD STORAGE & FRESHENING INST SHANXI ACADEMY OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, recently in Spain, Gandifruit, a product based on Candida sake, was registered. It is mainly used for the preservation of pome fruits. At present, there are few microorganisms that have the functions of preservation and preservation of berries, pome fruits and drupe fruits.

Method used

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  • Bacterial strain capable of antagonizing botrytis cinerea pathogens after picking of fruits and vegetables and application of bacterial strain
  • Bacterial strain capable of antagonizing botrytis cinerea pathogens after picking of fruits and vegetables and application of bacterial strain
  • Bacterial strain capable of antagonizing botrytis cinerea pathogens after picking of fruits and vegetables and application of bacterial strain

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Experimental program
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Effect test

Embodiment 1

[0032] Isolation and Screening of Bacillus Velez H-1 Strain

[0033] Botrytis cinerea, the pathogenic bacterium of tomato gray mold, was isolated from the surface of naturally occurring tomato fruits. After purification, it was inoculated on PDA medium. into 1×10 6 Spore mL -1 spore suspension. The pear fruits used to isolate antagonistic bacteria were collected from Yuluxiang Liyuan, Yuanping City, Shanxi Province; the cherry tomatoes used for inoculation were purchased from Xiaodian District, Taiyuan City.

[0034] Isolation of Bacillus Velez H-1 strain: Take pome fruits of 3 storage periods and put them into 0.2M phosphate buffer solution, rotate and shake in a shaker at 100r / min for 10min, discard the washing solution, and then Add phosphate buffer, wash in an ultrasonic cleaner for 30s, and perform a second wash. After the liquid obtained from the second washing was serially diluted 10 times, 100 times and 1000 times, 100 μL of each was spread on the PDA medium, and c...

Embodiment 2

[0038] Identification of Bacillus Velez H-1 Strain

[0039] 2.1 Cell culture

[0040] After re-culturing the lyophilized strains, they were separated by streaking on a plate, and after 20 hours of culture, a single colony was picked and inoculated into 20 ml of liquid medium, and cultured at 30°C and 200 rpm for 15 hours.

[0041] 2.2 Genomic DNA extraction

[0042](1) Take a single colony and inoculate it in 5ml of the corresponding culture medium, and cultivate it at a suitable temperature. Depending on the growth rate of the bacteria, the incubation time can range from a few hours to a few days.

[0043] (2) Take 1.0ml of bacterial liquid in a 1.5ml centrifuge tube, centrifuge at 13000rpm for 2min, and discard the supernatant.

[0044] (3) The pellet was resuspended in 1.0 ml of 0.85% NaCl.

[0045] (4) Centrifuge at 13,000 rpm for 2 minutes at room temperature, and discard the supernatant.

[0046] (5) The pellet was resuspended in 550 μl 1×TE.

[0047] (6) Add 17 μl...

Embodiment 3

[0093] Study on Antibacterial Effect of Bacillus Velez H-1

[0094] 1. In vitro inhibitory effect of Bacillus Velez H-1 on pathogenic bacteria

[0095] Purify Bacillus Velez H-1 by using the dish method, pick a single colony and inoculate it in 100mL of LB liquid medium, shake and culture at 200r at 30°C for 24h, measure the concentration by the dilution plate method, and inoculate it with sterile water The preparation concentration was 1×10 5 , 1×10 6 , 1×10 7 , 1×10 8 , 1×10 9 antagonistic antibiotic suspension. In a petri dish (diameter 9cm) equipped with PDA, after the medium is solidified, add 100 μL of antagonistic bacteria H-1 suspension of different concentrations, spread evenly, dry it with sterile air, and then insert it at the central point Botrytis cinerea mold cake with a diameter of 5mm, as a control, add sterile water. Cultivate at a constant temperature at 28°C, measure the diameter of the colony after 5-7 days, and calculate the inhibition rate. Each tr...

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Abstract

The invention belongs to the technical field of microorganisms and particularly relates to a bacterial strain capable of antagonizing botrytis cinerea pathogens after picking of fruits and vegetablesand application of the bacterial strain. The antagonistic bacterium is Bacillus velezensis H1, collected in the China Center for Type Culture Collection (CCTCC) with the collection number of CCTCC NO:M 2019274, is separated from pear fruit surfaces, has very significantly effects of inhibiting botrytis cinerea after picking of fruits and vegetables, has higher inhibition actions on botrytis cinerea, can remarkably inhibit germination of botrytis cinerea spores and growth of germ tubes, has good inhibition functions on the botrytis cinerea after picking of pears, natural rotting after picking of winter jujubes and natural l rotting after picking of grapes, and provides an environmental-friendly, simple and effective way for preservation of fruits and vegetables after picking.

Description

technical field [0001] The invention belongs to the technical field of microbes, and in particular relates to a bacterial strain antagonizing postharvest gray mold pathogens of fruits and vegetables and an application thereof. Background technique [0002] The fruit industry is the third largest industry in my country's planting industry after grain and vegetables, and occupies a very important position in the national economy. However, during the post-harvest storage and transportation of fruits and vegetables, due to pathogenic bacteria infection, respiration and aging, they are prone to rot and deterioration, resulting in huge losses. It is estimated that in developed countries, 20-25% of the fruits and vegetables are rotted due to the infection of pathogenic bacteria in the process of picking every year, and the loss is even more serious in developing countries. According to statistics, the loss caused by post-harvest rot in my country accounts for about 30% of the tota...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23B7/155C12R1/07
CPCC12N1/20A23B7/155C12R2001/07C12N1/205
Inventor 施俊凤杜静婷孙常青冯志宏焦旋白宇皓杨志国陈嘉高振峰陈会燕
Owner FARM PROD STORAGE & FRESHENING INST SHANXI ACADEMY OF AGRI SCI
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