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A kind of Paenibacillus polymyxa used for preventing and treating Verticillium wilt of crops and its application

A technology of Paenibacillus polymyxa and Bacillus, which is applied in the field of development and utilization of microbial germplasm resources, can solve the problems of great differences in antagonism effects, and achieve high-efficiency antagonism and obvious therapeutic effects

Active Publication Date: 2018-06-26
ZHEJIANG XINNONG CHEM CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the antagonistic effects of different species, even strains from different sources of the same species, vary greatly

Method used

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  • A kind of Paenibacillus polymyxa used for preventing and treating Verticillium wilt of crops and its application
  • A kind of Paenibacillus polymyxa used for preventing and treating Verticillium wilt of crops and its application
  • A kind of Paenibacillus polymyxa used for preventing and treating Verticillium wilt of crops and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 strain isolation and screening

[0030] The soil samples for testing were collected from the cotton disease nursery of the Hebei Academy of Agricultural Sciences and the cotton test field of the Northwest Agriculture and Forestry University in Sanyuan County, Shaanxi Province. The soil samples were randomly collected in sterile bags, sealed and stored in a refrigerator at 4 °C.

[0031] Screening of biocontrol bacteria in soil by double-layer plate method. Put the Verticillium dahliae cake that has been cultured for 4-5 days in sterile water, and prepare a concentration of 10 6 per mL of spore suspension. Take an appropriate amount (1g) of soil sample to prepare a soil suspension in sterile water, and serially dilute to 10 -2 -10 -3 . Double-layer plate method: evenly coat the prepared spore suspension of Verticillium dahliae on PDA, grow at 28°C for 48 hours, pour LB on the upper layer of PDA, and evenly coat the soil suspension. Place the plate under da...

Embodiment 2

[0043] The identification of embodiment 2 bacterial strains

[0044] The strain SX-3 with the best screening effect was identified.

[0045] Morphological identification

[0046] The colony surface of the strain on LB medium was smooth and milky white ( figure 1 ). Gram-positive bacteria were determined by staining test. Under the scanning electron microscope, it is a rod-shaped bacterium with a size of 0.58×2.9 μm ( figure 2 ).

[0047] 16S rDNA sequence analysis

[0048] The strain was inoculated in NA culture medium (yeast powder 1g / L, beef extract 3g / L, peptone 5g / L, sucrose 10g / L, pH=7.0), and after culturing at 30°C and 180rpm for 120 hours, DNA extraction was performed; 16S rDNA universal primers were used for PCR amplification to obtain an electrophoretic band of about 1400 bp, which was recovered and sequenced by Sangon Bioengineering Co., Ltd. (Shanghai). The determined 16SrDNA sequence was compared with the sequence in the GenBank database to determine its t...

Embodiment 3

[0056] Example 3 Fermentation of bacterial strain SX-3 and detection of antagonistic components

[0057] The culture medium can be LB liquid medium, its formula is: tryptone 10g / L, yeast extract powder 5g / L, sodium chloride 10g / L, adjust the pH to 7.0-7.2.

[0058] 50L automatic fermenter culture: Inoculate Paenibacillus polymyxa in LB culture medium and cultivate to culture medium OD 600 0.6 to 0.8 to obtain the seed liquid; inoculate the seed liquid into a 50L fermenter, the expansion culture temperature should be between 28 and 35 °C, the culture time should be between 48 and 72 hours, the aeration ratio is 0.4 to 2:1, and the rotation speed 300-800rpm, pH should be between 6-7.5. The fermentation medium is: first gelatinize starch, and then add yeast powder, protein powder, glucose, MgSO 4 、KH 2 PO 4 , CaCO 3 medium, sterilized at 121°C for 30 minutes. The final bacterial concentration can usually reach 1×10 10 ~1×10 11 CFU / mL.

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Abstract

The invention discloses paenibacillus polymyxa for preventing and treating vertieillium wilt in crops and application of the paenibacillus polymyxa. The strain is named as paenibacillus polymyxa SX-3, with preservation number as CGMCC 11638. The invention provides an inoculant containing the paenibacillus polymyxa. The strain of the invention has an inhibitory effect on both spore germination and mycelial growth of verticillium dahlia and the strain has an efficient antagonistic action. The strain of the invention is capable of preventing and treating crop verticillium wilt caused by the verticillium dahlia, the strain has a significant effect on preventing and treating cotton verticillium wilt and control efficacy of pot experiments reaches 86% or above; and bacterium liquid containing the strain of the invention, when irrigated to the roots of crops in a late disease period, can still achieve a significant therapeutic effect; therefore, the strain has a great potential in the application to bio-pesticides.

Description

technical field [0001] The invention belongs to the field of development and utilization of microbial germplasm resources, and in particular relates to a Paenibacillus polymyxa used for preventing and treating Verticillium wilt of crops caused by Verticillium dahliae and application thereof. Background technique [0002] Verticillium dahliae (Verticillium dahliae) belongs to Ascomycota fungus, and it mainly overwinters with microsclerotia on soil, diseased tissues, infected seeds and unripe soil miscellaneous fertilizers, and can harm 660 species of plants in 38 families, among which Field crops include cotton, sunflower, eggplant, pepper, tomato, tobacco, potato, etc., and the disease caused by it is called Verticillium wilt. After overwintering, the microsclerotium germinates and produces a large number of conidia. The conidia germinate and invade root hairs, root epidermal cells or root wounds, enter the vessel through the cortex, destroy the vascular system of the plant,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/00A01P3/00A01G13/00C12R1/01
Inventor 张敬泽李晓林祝水金
Owner ZHEJIANG XINNONG CHEM CO LTD
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