Bio-control combined bacterium agent CB28 admixture for preventing cucumber downy mildew
A technology of cucumber downy mildew and compound bacterial agent, which is applied in the direction of biocides, bactericides, biocides, etc., can solve the problems of high cost, high pesticide residues, and difficult control of cucumber downy mildew, and achieve low price, Effect of cost saving and protection of farmland ecosystem
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Embodiment 1
[0030] Strain source: BS13 was isolated from banana rhizosphere soil in Machong, Dongguan City, Guangdong Province; BJN15 was isolated from banana pseudostem in Machong, Dongguan City, Guangdong Province.
[0031] 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.)
[0032] Screening basis for strains: Determination of five enzyme activities in hydrolysis (protease, chitinase, cellulase, siderophilic, IAA) 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 determination of antagonistic activity against Phytophthora bacterium (confrontation culture method, Lin Fucheng, Li Debao. Lysis of Bacillus subtilis to plant pathogenic fungi[J]. Phyto...
Embodiment 2
[0039] A biocontrol complex bacterial agent CB28 mixture for controlling cucumber downy mildew, which contains two bacterial components: Bacillus megaterium BS13 (CGMCC NO.4734) and Enterobacter BJN15 (CGMCC NO.4733).
[0040] The preparation method of the biocontrol compound bacterial agent CB28 mixture of this control cucumber downy mildew is as follows:
[0041] (1) Cultivation of seed bacteria liquid: respectively inoculate Bacillus megaterium BS13 with the preservation number of CGMCC NO.4734 and Enterobacter BJN15 with the preservation number of CGMCC No.4733 into the LB culture medium, at 30-32°C, 180- Cultivate under the condition of 220rpm until the OD value at 600nm is between 0.5 and 0.8, and end the cultivation to obtain the seed bacterial liquid;
[0042] (2) Preparation of wet bacteria: Take the seed bacteria solution in step (1), add it to LB culture solution at a volume ratio of 1:250, shake and culture at 30-32°C, 180-220rpm for 45-48 hours, and then centrifug...
Embodiment 3
[0047] Example 3 Indoor sporangia germination test
[0048] Method: slide culture method (reference: NY / T1156.1-2006 Pesticide Indoor Bioassay Test Guidelines Fungicide Part 1: Inhibition of pathogenic fungus spore germination test concave slide method): CB28 prepared according to the method of Example 2 Mixture bacterial solution was prepared to the corresponding concentration, and was added dropwise on the sterilized glass slides respectively, and then 10 μl of prepared downy mildew sporangia suspension was added dropwise to the bacterial solution, the concentration was 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 length of the germ tube is greater than 1 / 2 of the sporangia width of downy mildew fungus, and the calculation formu...
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