Bacillus amyloliquefaciens strain and application thereof in prevention and treatment of apple continuous cropping obstacles

A technology of amyloliquefaciens and bacilli, applied in the field of agricultural microorganisms

Active Publication Date: 2021-01-26
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few reports on Bacillus amyloliquefaciens used to control apple cropping obstacles

Method used

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  • Bacillus amyloliquefaciens strain and application thereof in prevention and treatment of apple continuous cropping obstacles
  • Bacillus amyloliquefaciens strain and application thereof in prevention and treatment of apple continuous cropping obstacles
  • Bacillus amyloliquefaciens strain and application thereof in prevention and treatment of apple continuous cropping obstacles

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Example 1: Isolation and identification of bacterial strains

[0068] 1. Isolation and purification of strains:

[0069] Rhizosphere bacteria were isolated by a modified dilution spread plate method. Fresh soil samples were passed through a sieve with a diameter of 3-4 mm to remove impurities in the soil. Weigh 5 g of rhizosphere soil, add it to a triangular flask (with glass beads) filled with 45 mL of sterile water, vibrate at 180 rpm for 30 min, let it stand for 5 min, and serially dilute to 1×10 with sterile water. -4 100 μL of the supernatant of the gradient dilution was spread on the LB plate, repeated three times, and cultured upside down in a constant temperature incubator at 28°C. After a single colony grows in the plate, pick a single colony of different shapes and streak on a new LB plate for purification. At the same time, the purified strain was inoculated in liquid LB medium, shaken at 180rpm at 28°C for 12h, centrifuged at 10000r at 4°C for 10min, disc...

Embodiment 2

[0093] Embodiment 2: the broad-spectrum antibacterial effect of bacterial strain QSB-6

[0094] In order to further verify whether the QSB-6 strain has a broad-spectrum antibacterial effect on plant-derived diseases, another four common pathogenic fungi were selected for further antibacterial tests. The results are shown in Table 2.

[0095] The results of confrontation test showed that QSB-6 strain had a strong inhibitory effect on the growth of four kinds of pathogenic Fusarium mycelium. It also has a strong inhibitory effect on Penicillium citrus, Grape Anthracnose, Alternaria corn and Apple rot. Therefore, the strain QSB-6 has a broad-spectrum antibacterial effect on plant pathogenic bacteria under the conditions of the in vitro petri dish test.

[0096] Table 2: Inhibition effect of bacterial strain QSB-6 on 8 kinds of pathogenic bacteria

[0097]

[0098] Note: –, no zone of inhibition; +, zone of inhibition 10mm. Different lowercase letters indicate significant ...

Embodiment 3

[0099] Embodiment 3: the optimization of bacterial strain QSB-6 fermentation condition

[0100] 1. Test method:

[0101] The preserved strains were transferred to LB solid medium and cultured at 37°C for 24 hours. Pick the activated single colony and inoculate it into the Erlenmeyer flask containing 50ml LB liquid medium, 160r min -1 , 37 ° C shaker shaking culture until OD600 = 1 or so for use. The seed liquid is connected to the Erlenmeyer flask containing the culture medium according to the inoculation amount of 2%; at 200r·min-1 , Cultivate in a shaker at 37°C for 24h.

[0102] Fermentation culture studies with single factor experiments.

[0103] 1.1 Carbon source: use 20g·L respectively -1 Equal amounts of sucrose, maltose, glucose, lactose and soluble starch replaced the carbon source of the basal culture medium. After screening different concentrations of 0.1%, 0.5%, 1%, 1.5% and 2%, use shake flask fermentation culture, after 24h, determine the optimal carbon sour...

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Abstract

The invention discloses a bacillus amyloliquefaciens strain and application thereof in prevention and treatment of apple continuous cropping obstacles. The bacillus amyloliquefaciens strain is preserved in China Committee for Culture Collection of Microorganisms under the preservation number of CGMCC NO. 15307 since January 25, 2018. The bacillus amyloliquefaciens strain provided by the inventionhas an inhibitor effect on various pathogenic bacteria, and is extremely strong in antagonistic effect on fusarium oxysporum, fusarium verticillatum, fusarium proliferatum and fusarium solani which may cause apple continuous cropping obstacles. The bacillus amyloliquefaciens strain is also capable of promoting growth of overground parts and underground parts of continuous cropping apple tree seedlings, increasing bacterium content in soil, raising bacterium and fungus ratios, and improving activity of soil urease, phosphatase, sucrase and catalase in apple continuous cropping soil; and thus, the bacillus amyloliquefaciens strain has excellent effect in relieving apple continuous cropping obstacles, and can be used for preventing and treating apple continuous cropping obstacles.

Description

technical field [0001] The invention relates to the technical field of agricultural microorganisms, in particular to a strain of Bacillus amyloliquefaciens and its application in preventing and treating apple cropping obstacles. Background technique [0002] my country's apple cultivation area, total output, per capita share and export volume all rank first in the world, and China has become the world's largest apple producer and consumer. At present, my country's apple cultivation area is 37 million mu, and the apple output is about 43.8 million tons. However, the apple orchards in these traditionally advantageous cultivation areas were mainly established in the 1980s and early 1990s, and most of them have entered the aging period. In Shandong Province alone, there will be nearly 67,000 to 100,000 hm in the next five years 2 Apple orchards face renewal. Due to the limited land resources, when renewing old orchards, apple tree replanting cultivation cannot be avoided, resul...

Claims

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Application Information

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IPC IPC(8): C12N1/20C05F17/20C05G3/60A01N63/22A01P3/00C12R1/07
CPCA01N63/22C05F3/00C12N1/20C05F17/20C05G3/60C12N1/205C12R2001/07C05F11/00C05F11/08Y02W30/40
Inventor 尹承苗段亚楠毛志泉刘鑫陈学森沈向
Owner SHANDONG AGRICULTURAL UNIVERSITY
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