Application of Bacillus velezensis NZ-4 to promotion of vegetation

A technology that promotes plant growth and bacillus, applied in the fields of microbiology and plant disease biocontrol, can solve the problems of potato wilt aggravation, restricting 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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  • Application of Bacillus velezensis NZ-4 to promotion of vegetation
  • Application of Bacillus velezensis NZ-4 to promotion of vegetation
  • Application of Bacillus velezensis NZ-4 to promotion of vegetation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Isolation and Purification of Potato Fusarium Wilt Antagonist NZ-4

[0025] 1 Materials and methods

[0026] 1.1 Soil sample collection

[0027] 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.

[0028] 1.2 Tested target pathogenic bacteria

[0029] 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).

[0030] 1.3 Test medium

[0031] LB medium for bacteria and PDA medium for fungi.

[0032] 1.4 Isolation and purification of Bacillus strains

[0033] 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

[0042] Identification of embodiment 2 antagonistic strains

[0043] The screened antagonistic strains were identified by colony morphology, Gram staining and molecular biology.

[0044] Sequence Analysis of the gyrB Gene: gyrB Gene Amplification Primer Sequences [15] 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.

[0045] On the LB medium plate, the colony ...

Embodiment 3

[0047] Pot experiment of embodiment 3 antagonistic bacteria preventing and treating potato wilt

[0048] 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.

[0049] Disease index: (DI) = ∑ (disease level ...

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Abstract

The invention provides application of Bacillus velezensis NZ-4 to promotion of vegetation. The fermentation broth of the bacterial strain NZ-4 can substantially promote the growth of potato plants. The Bacillus velezensis NZ-4 and a microbial strain thereof are of great significance to prevention and treatment of soil borne diseases and successive cropping obstacles in the farmland of China, to sustainable yield increase of grain and to the protection of the ecological environments of farmland.

Description

technical field [0001] The invention relates to the technical field of microbiology and plant disease biocontrol, in particular to the application of Bacillus veleisi NZ-4 in promoting plant growth. 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 croppin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/00A01P21/00C12R1/07
CPCA01N63/00C12N1/205C12R2001/07
Inventor 张岱朱杰华杨志辉赵冬梅李彩虹杨毅清潘阳丁明亚
Owner HEBEI AGRICULTURAL UNIV.
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