Bacillus amyloliquefaciens for preventing and curing tomato bacterial wilt and application of bacillus amyloliquefaciens
A technology for dissolving starch spores and bacilli is applied in the field of crop disease prevention and smelting, and can solve problems such as unstable effect, food safety, and environmental pollution.
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Embodiment 1
[0023] Example 1 The acquisition of biocontrol strain W118 for preventing and treating tomato bacterial wilt
[0024] Screening and Isolation of Bacillus amyloliquefaciens
[0025] (1) Soil samples were collected from the rhizosphere soil of healthy plants in fields with severe tomato bacterial wilt disease. The healthy plants were uprooted and shaken gently. After most of the soil was shaken, the remaining soil close to the roots was collected together with some fibrous roots. Pack it in a sealed bag and store it in a 4°C refrigerator for later use.
[0026] (2) Add 10 g of rhizosphere soil of healthy tomato plants into a conical flask filled with 90 mL of sterile water, vibrate vigorously on a shaker at 30° C. for 30 min, and let stand for 30 min;
[0027] (3) The supernatant was serially diluted and spread on the modified medium of beef extract peptone, each gradient was repeated 3 times, and after culturing at 30°C for 24 hours, the medium was filled with 1.0×10 8 The th...
Embodiment 2
[0030] Example 2 Identification of biocontrol strain W118 for preventing and treating tomato bacterial wilt
[0031] The strain W118 was inoculated on beef extract peptone modified medium, cultured at 30°C for 48 hours, the genomic DNA of the antagonistic strain was extracted with a bacterial genomic DNA extraction kit, and amplified with bacterial 16S rRNA primers. (F: 5'-AGAGTTTGATCCTGGCTCAG-3'R: 5'-TACCTTGTTACGACTT-3'). The reaction system is: Mg 2+ 2.5μl, 10×buffer 2.5μl, dNTP (10mmol·L -1 ) 2 μl, upstream and downstream primers (10mmol L -1 ) each 1 μl, template DNA 1 μl, Taq enzyme (5U·μl -1 )0.3μl, ddH 2 O 14.7 μl, the total system is 25 μl. All reagents used were purchased from Dalian Bao Biological Engineering Co., Ltd.
[0032] PCR reaction conditions: pre-denaturation at 95°C for 5min, denaturation at 95°C for 30s, annealing at 53°C for 1min, extension at 72°C for 2min, holding at 72°C for 10min, and a total of 30 cycles. The PCR product was detected by 1% (w...
Embodiment 3
[0035] Example 3 Antagonism of Bacillus amyloliquefaciens W118 to tomato bacterial wilt pathogen
[0036] Spot the purified strain W118 on beef extract peptone modified medium with a sterile toothpick, culture at 30°C for 12h, and use 1.0×10 8cfu / m of R. solanacearum pathogen liquid spray liquid, measure the diameter of the inhibition zone after 48h, repeat 3 times, observe the size of the inhibition zone, by the attached figure 1 and 2 It can be seen that Bacillus amylobacter W118 has a stronger inhibitory effect on tomato R. solanacearum.
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