Biological prevention bacterial strain BS1 (bacillus cereus) for preventing and controlling greenhouse cucumber downy mildew and application thereof
A cucumber downy mildew, greenhouse technology, applied in the direction of application, bacteria, biocide, etc., to achieve the effect of saving expenses and increasing income
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Embodiment 1
[0024] Strain source: BS1 was isolated from banana rhizosphere soil in Mayong, Dongguan City, Guangdong Province.
[0025] Strain Isolation Method: Plate Dilution Method (Microbiological Laboratory of Nanjing Soil Institute, Chinese Academy of Sciences. Soil Microbial Research Method [M]. Beijing: Science Press Du, 1985.)
[0026] Screening basis for strains: detection of the activity of five enzymes in the strain hydrolysis (protease, chitinase, cellulase, siderophilic, IAA determination of Yang J.-H., Liu H.~X., Zhu G-M., Xu L. .-P., Pan Y.-L., and Guo J.-H.*.2008.Diversity analysis of antagonists from rice associated bacteria and their application in biocontrol of rice diseases.Journal of Applied Microbiology.104(1): 91~104.) and antagonism against Phytophthora (confrontation culture method, Lin Fucheng, Li Debao. Lysis of Bacillus subtilis to plant pathogenic fungi [J]. Phytopathology Acta, 2003, 33( 2): 174-177.), through the evaluation of bacteria (Faltin et al., 2004),...
Embodiment 2
[0031] Embodiment 2 bacterial agent preparation
[0032] (1) Cultivation of seed bacteria liquid: Inoculate BS1 (CGMCC NO.4736) into LB culture liquid, culture at 32°C and 200 rpm for 18-24 hours, and after 16 hours, take samples at 600nm every 3 hours in the ultra-clean workbench. OD value, during the measurement process, use the uninoculated culture solution to adjust to zero, until the OD value of the seed bacteria solution is between 0.5 and 0.8, the culture is ended, and the seed bacteria solution is obtained;
[0033] (2) Preparation of wet bacteria: Add the seed bacteria solution into a fermenter equipped with 200L fermentation broth at a volume ratio of 1:100, shake and cultivate at 250 rpm at 30°C for 20 to 24 hours, maintain the pH value of the fermentation broth at 7.0, and the dissolved oxygen 20%, ventilation volume 6m 3 / hour, tank pressure 0.05 ~ 0.1KPa. Then 6000rpm centrifugal 10min, every 1000ml culture solution obtains 12~15g wet bacteria, gets wet bacter...
Embodiment 3
[0035] Example 3 Indoor sporangia germination test
[0036]Method: slide culture method (references: NY / T 1156.1-2006 Pesticide Indoor Bioassay Test Guidelines Fungicide Part 1: Inhibition of pathogenic fungus spore germination test concave slide method): the bacterial strain BS1 ( CGMCC NO.4736) is that the biocontrol bacterium agent of effective bacterium is formulated into corresponding concentration, is dripped respectively on the sterilized glass slide, then drops 10 μ l downy mildew sporangia suspension prepared in the bacterial liquid, concentration is 10 5 sporangia / ml, 4 repetitions for each treatment, with sterilized water as a control, placed in a light incubator at 25°C for 48 hours to observe the germination rate of sporangia, microscopic examination was based on 30 sporangia in the field of vision, and observed The standard is that the germ tube length is greater than 1 / 2 of the sporangia width of downy mildew bacteria, and the calculation formula is inhibition ...
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