Application of Bacillus Velez nz-4 in promoting plant growth
A technology of bacillus and strains, applied in the fields of microbiology and plant disease biocontrol, can solve the problems of aggravating potato wilt and restricting the development of potato industry, and achieve the effect of promoting growth
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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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