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Antagonistic bacterium NZ-4 for wilt of potatoes and its application

A technology of antibacterial and NZ-4 against fusarium wilt, which is applied in the fields of microbiology and plant disease biocontrol, can solve the problems of aggravation of potato fusarium wilt, restrict the development of potato industry, etc., and achieve the effect of promoting growth

Active Publication Date: 2018-07-31
HEBEI AGRICULTURAL UNIV.
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, with the implementation of the potato staple food strategy, the potato planting area has expanded year by year, and the resulting continuous cropping has led to the increasing severity of potato wilt, which seriously restricts the development of the potato industry.

Method used

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  • Antagonistic bacterium NZ-4 for wilt of potatoes and its application
  • Antagonistic bacterium NZ-4 for wilt of potatoes and its application
  • Antagonistic bacterium NZ-4 for wilt of potatoes and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Isolation and Purification of Potato Fusarium Wilt Antagonist NZ-4

[0026] 1 Materials and methods

[0027] 1.1 Soil sample collection

[0028] The rhizosphere soil of healthy plants was collected from the potato wilt disease-infected fields in Baoding, Chengde, Zhangjiakou, Qinhuangdao, Hebei, and Inner Mongolia and Guizhou. Temporarily stored at 4°C for the isolation of antagonistic bacteria.

[0029] 1.2 Tested target pathogenic bacteria

[0030] One Fusarium oxysporum (Y10-2) strain, which is the dominant strain of Fusarium oxysporum, was provided by our laboratory (Plant Pathology Laboratory of Hebei Agricultural University).

[0031] 1.3 Test medium

[0032] LB medium for bacteria and PDA medium for fungi.

[0033] 1.4 Isolation and purification of Bacillus strains

[0034] Bacillus was isolated by heat-treated soil dilution separation method, continuously streaked and cultured until pure, mixed in 70% glycerol at a ratio of 1:1, and stored in a -...

Embodiment 2

[0043] Identification of embodiment 2 antagonistic strains

[0044] The screened antagonistic strains were identified by colony morphology, Gram staining and molecular biology.

[0045] Sequence analysis of gyrB gene: gyrB gene amplification primer sequence forward primer gyrB-F: 5'-TTGRCGGHRGYGGHTATAAAGT-3'; reverse primer gyrB-R: 5'-TCCDCCSTCAGARTCWCCCTC-3'. The PCR amplification program was: pre-denaturation at 95°C for 3 min; denaturation at 94°C for 30 s, annealing at 55°C for 45 s, extension at 72°C for 1 min, 35 cycles; final extension at 72°C for 10 min, and termination at 4°C. After the PCR product was detected on 1.5% agarose gel electrophoresis, it was sent to Sangon Bioengineering (Shanghai) Co., Ltd. for sequencing. The measured sequences were compared by BLAST in the NCBI database, and the phylogenetic analysis of the isolated Bacillus and model bacteria was carried out using Clustalx and MEGA6.0 software.

[0046] On the LB medium plate, the colony of strain N...

Embodiment 3

[0050] Pot experiment of embodiment 3 antagonistic bacteria preventing and treating potato wilt

[0051] Use 1.0×10 root dipping method 8 cfu / mL antagonistic bacteria fermentation broth was treated for 2 hours, and water treatment was used as the control, with 30 plants per treatment, a total of 60 plants; after 24 hours, 1 × 10 6 Each / mL of Fusarium oxysporum Y10-2 spore suspension soaked the roots for 30 minutes, and the potato plants inoculated with the pathogen were placed in the dark for 24 hours, and the ambient humidity was kept at 95% to promote the infection of the pathogen. After 24h, they were transferred to normal light conditions (14h light: 10h dark) for cultivation. After 20 days, the potato disease was investigated, and the diseased plants were isolated and identified by the conventional tissue isolation method. The disease index was calculated according to the pathogenicity grading standard of Krumholz et al.

[0052] Disease index: (DI) = ∑ (disease level ...

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Abstract

The invention provides an antagonistic bacterium NZ-4 for wilt of potatoes. The antagonistic bacterium NZ-4 is Bacillus velezensis, with an accession number of CGMCC No. 13606. The invention also provides the application of the antagonistic bacterium NZ-4 to prevention and treatment of soil-borne diseases of crops. Pot experiment results show that the antagonistic bacterium NZ-4 has significant prevention and treatment effect on wilt of potatoes, with prevention and treatment effect being 62.14%. In addition, the strain NZ-4 also has strong antagonistic effect on pathogens leading to anthracnose of scab potatoes, root rot of wheat, etc. The application of the fermentation broth of the antagonistic bacterium NZ-4 can significantly promote the growth of potato plants. The antagonistic bacterium NZ-4 and microbial agents thereof are of great significance for prevention and treatment of the soil-borne diseases and continuous cropping obstacles of farmland in China, the sustainable yield increase of grain and the protection of farmland ecological environment.

Description

technical field [0001] The invention relates to the technical field of microbiology and plant disease biocontrol, in particular to potato fusarium wilt antagonist NZ-4 and application thereof. Background technique [0002] Potato wilt is a soil-borne disease of potato caused by Fusarium oxysporum. It is widely distributed and occurs in various planting areas in my country. Potato Fusarium wilt can cause damage to the entire growth period of the plant. The pathogenic bacteria mainly invade from the root wound and root hair growth point of the plant, and then destroy the vascular bundles of the roots, stems and leaves, causing part or the entire vascular bundles to turn brown, allowing the plant to absorb and the loss of water transport function, eventually causing the plant to wither and die. In recent years, with the implementation of the potato staple food strategy, the potato planting area has expanded year by year, resulting in continuous cropping and heavy cropping, whi...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/00A01P1/00A01P3/00C05G3/00C12R1/07
CPCA01N63/00C05D1/00C05F11/08C05G3/00C12N1/20C12N1/205C12R2001/07C05G1/00
Inventor 杨志辉朱杰华张岱赵冬梅李彩虹潘阳杨毅清丁明亚
Owner HEBEI AGRICULTURAL UNIV.
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