A Strain of Bacillus siamese and Its Application
A technology of Bacillus and strains, applied in Bacillus siamese and its application fields, can solve the problems of enhanced drug resistance, harmfulness, and chemical control can not be used as the most suitable means, to achieve growth promotion, broad application prospects, and strong control effect of effect
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Embodiment 1
[0076] Example 1 Identification of bacterial strain LZ88
[0077] Using the bacterial strain LZ88 genomic DNA as a template, the bacterial 16S rDNA universal primers 27F (5'-AGAGTTTGATCMTGGCTCAG-3', SEQ ID NO.1) and 1492R (5'-TACGGYTACCTTGTTACGACTT-3', SEQ ID NO.2) were used to amplify 16S rDNA sequence. Amplification system and procedure: 25 μL of PCR system contains 5.0 μL of 5×PCR buffer, DNA template (100 ng μL -1 ) 1.0 μL, dNTPs (10 mM) 0.75 μL, forward primer (10 μM) 0.75 μL, reverse primer (10 μM) 0.75 μL, MgCl 2 (25mM) 2.5μL, Taq DNA polymerase (5UμL -1 Promega) 0.25μL, the rest ddH 2 O make up. The PCR amplification program was pre-denaturation at 94°C for 3min; denaturation at 94°C for 30s, annealing at 55°C for 30s, extension at 72°C for 1min30s, and 30 cycles. Finally, extend at 72°C for 10 min and store at 4°C. After the reaction, the PCR products were analyzed by agarose gel electrophoresis. The obtained fragments were subjected to sequencing analysis. T...
Embodiment 2
[0079] Example 2 Bacillus siamensis (Bacillus siamensis) LZ88 antibacterial ability assay
[0080] The LZ88 strain was isolated from the rhizosphere soil of tobacco, and its antibacterial activity against tobacco red spot was determined by the confrontation culture method. The activated pathogenic bacteria cake was obtained with a sterilized puncher, and the pathogenic bacteria cake was first inoculated on fresh PDA. In the center of the plate, the antagonistic bacteria point was placed at a distance of 2.5 cm from the bacterium cake, and the plate only inoculated with the bacterium cake was used as a control. Place them in a 28°C incubator upside down. After 5 days, the colony diameters of the treatment and control were measured, and the bacteriostatic rate was calculated. The results showed that strain LZ88 was effective against tobacco red spot (Alteraria alternata), tobacco black shank (Phytophthora parasitica var. sp.cucumebrium Owen) and peanut sickle disease (Fusarium...
Embodiment 3
[0081] The mensuration of the antibacterial activity of embodiment 3 volatile antibacterial substances
[0082]The activity of volatile substances was determined by the plate buckle method. Inoculate the cake of Alternaria Alternaria in the center of the PDA plate, and inoculate the strain LZ88 on the NA plate at the same time, buckle the mouths of the two petri dishes together, and seal them with a parafilm. Cultivate in a constant temperature incubator at 28°C. After 7 days, observe the growth of the control and treatment. When cultivating, it should be noted that the plate inoculated with pathogenic bacteria should be placed at the top, and the plate inoculated with antagonistic bacteria LZ88 should be placed at the bottom.
[0083] The experimental results show that on the plate, the volatile substances of the antagonistic strain LZ88 can effectively inhibit the growth of Alternaria tabacum. In the control, the Alternaria has covered the entire plate, and in the experimen...
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