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Application of a sphingobacterium hotanense in the control of parasitic nematodes

A nematode and microorganism technology, applied in the field of microorganisms, can solve the problems of limited application of biocontrol preparations, biocontrol bacterial strain selection, lag in production technology and industrial production research, and achieve the effect of inhibiting root-knot nematodes

Active Publication Date: 2021-10-08
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, compared with biocontrol fungi, the research on the strain selection, production process and industrial production of biocontrol bacteria is relatively lagging behind, resulting in very limited application of biocontrol agents represented by biocontrol bacteria
[0005] Sphingobacterium is a kind of Gram-negative bacteria that does not produce spores widely in nature, and a variety of Sphingobacteria have been found to have degradative properties, including petroleum, pentachlorophenol, etc. However, regarding sphingosine There are few reports on the control of plant parasitic nematodes by Bacillus

Method used

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  • Application of a sphingobacterium hotanense in the control of parasitic nematodes
  • Application of a sphingobacterium hotanense in the control of parasitic nematodes
  • Application of a sphingobacterium hotanense in the control of parasitic nematodes

Examples

Experimental program
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Effect test

Embodiment 1

[0036] Isolation, Identification and Preservation of AMCC101218 Strain

[0037] (1) Isolation and screening process of AMCC101218 strain

[0038] An insecticidal strain was screened from plant rhizosphere soil, numbered AMCC101218.

[0039] (2) Identification of AMCC101218 strain

[0040] ①Main biological characteristics of AMCC101218 strain: the colony is round in shape with irregular edges and opaque, the color of the colony is yellow, and Gram staining is negative.

[0041] ②The main physiological and biochemical characteristics of the AMCC101218 strain were determined by BIOLOG 96 microwell plate, and the results of the physiological and biochemical characteristics of AMCC101218 are shown in Table 1:

[0042] Table 1

[0043]

[0044] ③Molecular biological characteristics of AMCC101218 strain

[0045] The molecular biology identification test method of AMCC101218 strain is carried out according to "6.5" in NY / T 1736-2009; MEGA6.0 is used to build the tree, and the r...

Embodiment 2

[0049] Determination of nematode killing ability of AMCC101218 strain

[0050] Use an inoculation loop to pick the AMCC101218 strain preserved on the slant and streak it on the LB (0.5% yeast extract, 1% peptone, 1% sodium chloride, 1.6% agar, pH7.0, 121 ° C, 20min autoclaving) plate for activation; The activated strains were inoculated in BPY culture solution, cultured on a shaker at 200 rpm at 37°C for 48 hours, centrifuged at 12000 rpm for 10 minutes, and the supernatant was collected.

[0051] The BPY fermentation medium includes the following components per liter: beef extract 5.0g, peptone 10.0g, yeast extract 5.0g, glucose 5.0g, NaCl 5.0g, balance water, pH 7.0.

[0052] Experiment on killing root-knot nematode incognita / root-knot nematode elephant ear:

[0053] Wash the picked root-knot nematode / meloidogyne elegans oocysts, put them in 0.5% sodium hypochlorite solution for disinfection for 3 minutes, rinse them with sterile water for 3 times, pick the oocysts and pl...

Embodiment 3

[0060] Preparation of AMCC101218 bacterial agent

[0061] (1) Strain activation: the AMCC101218 strain was streak-inoculated on an LB solid medium plate, and cultured at 37° C. for 24 hours.

[0062] (2) Seed solution preparation: the activated AMCC101218 was inoculated into a Erlenmeyer flask filled with LB liquid medium, and cultured in a constant temperature shaker at 37° C. and 180 rpm for 18 hours.

[0063] (3) Seed liquid expansion cultivation: Inoculate the seed liquid in step (2) into the seed tank liquid medium with an inoculation amount of 8%, the expansion medium is LB liquid medium, and cultivate at 37° C., 200 rpm for 8 hours.

[0064] (4) Fermentation culture: the seed liquid that expands culture in the step (3) is inoculated to BPY fermentation medium with the inoculum size of 8%, in 37 ℃, 200rpm cultivates 48h, obtains AMCC101218 liquid bacterial agent, viable count 1.7 * 10 8 CFU / mL, filled into 1L plastic bottle with cap.

[0065] The BPY fermentation mediu...

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Abstract

The invention discloses the application of a strain of Sphingobacterium hotanense in preventing and treating parasitic nematode diseases. The Sphingobacterium hotanense AMCC 101218 provided by the present invention has a preservation number of CGMCC No.19217; this bacterium can effectively control cucumber root-knot nematode disease, and its oocyst number, root-knot number, and insect population density have decreased by 71.28% compared with the blank control , 75.86% and 63.80%; AMCC101218 genome size is 4490254bp, GC content is 41.21%.

Description

technical field [0001] The invention relates to a Sphingobacterium hotanense and its application in preventing and treating plant parasitic nematode diseases, belonging to the technical field of microorganisms. Background technique [0002] With the promotion of protected cultivation, the problems of single crop species, regionalized planting, and high agricultural multiple cropping index have caused a major problem in the sustainable development of agriculture-continuous cropping obstacles. Continuous cropping obstacle refers to the continuous planting of the same crop in the same field under normal cultivation management measures, resulting in reduced yield, poor quality, frequent occurrence of diseases and insect pests, etc. Continuous cropping obstacles usually cause deterioration of soil physical and chemical properties, massive accumulation of phenolic compounds, uneven distribution of soil nutrients, secondary salinization of soil, and changes in soil enzyme activitie...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/20A01P5/00C12R1/01
CPCA01N63/00C12N1/20C12N1/205C12R2001/01
Inventor 周波王建宇张晓春王冰林榕姗赵鹏李根于丰源
Owner SHANDONG AGRICULTURAL UNIVERSITY
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