Sugarcane endophytic bacillus and application thereof
A technology for bacilli and amylolytic spores, applied in the field of biological control, can solve problems such as unsatisfactory effect and bacterial degradation, and achieve the effects of good development and application value, strong inhibitory effect, and elimination of pathogenic risks.
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Embodiment 1
[0042] Example 1 Isolation, purification and preservation of Bacillus amyloliquefaciens CGB15 strain in sugarcane leaves
[0043] 1. Preparation of LB medium
[0044]Weigh 10 g of tryptone (Tryptone, Oxoid LTD LP0042, England), 5 g of yeast extract (Yeasextract, Oxoid LTD LP0021, England), 10 g of sodium chloride (NaCl, Sinopharm Chemical Reagent Co., Ltd., 10019318), add 1000 mL of water and stir well , plus 15g of agar, fully heated to dissolve, subpackaged, sterilized at 121°C for 20min, and stored for later use.
[0045] 2. Isolation and purification of Bacillus amyloliquefaciens in sugarcane leaves
[0046] (1) Leaf collection:
[0047] The sugarcane leaves used for separation were collected in November 2017 from the sugarcane field of researcher Shen Wankuan, School of Agriculture, South China Agricultural University, Guangzhou, and roughly divided into young leaves, mature leaves, old Three groups of leaves.
[0048] (2) Leaf disinfection: wash 3 times with steriliz...
Embodiment 2
[0058] Embodiment 2 confrontation culture method measures the antifungal activity of Bacillus amyloliquefaciens CGB15 bacterial strain
[0059] 1. Preparation of culture medium
[0060] (1) Preparation of LB liquid medium: except that agar is not added, other formulas are the same as those of LB solid medium in Example 1, and the weighed formula medicament is fully dissolved and subpackaged in conical flasks (100mL culture fluid in each bottle) ), stoppered, bandaged, sterilized at 121°C for 20 minutes, cooled and stored for later use.
[0061] (2) Preparation of carrot culture medium: take 200g carrots, filter with 16 layers of gauze after fully squeezing the juice with a juice extractor, add water to 1000mL with the filtrate, add 15g agar, heat and dissolve it, and then pack it into conical flasks ( 100mL culture medium per bottle), sterilized at 121°C for 20min, cooled and stored for later use.
[0062] (3) Preparation of PDA medium: PDA broth (purchased from Dingguo Chan...
Embodiment 3
[0075] Example 3 Control Effect of Bacillus amyloliquefaciens CGB15 on Rice Blast
[0076] 1. The preparation of LB medium is the same as in Example 1; the preparation of CM and PDA medium is the same as in Example 2.
[0077] 2. The preparation of the bacillus plate bacteria suspension and the crude extract is the same as in Example 2.
[0078] 3. Select barley leaves and rice leaves with uniform growth, take the rice blast bacteria cake and cover them on the leaves, set up the clear water group and the fermentation broth group (the extraction process is the same as step 3 in Example 2), and add 5 μL to the bacteria in each group. Between the cake and the leaves. After cultivating in the dark at 28°C for one day, transfer to a 28°C light (12h dark: 12h light) incubator for 3 days, observe the disease and record it, the results are as follows: Figure 6 shown.
[0079] 4. Result analysis
[0080] Depend on Figure 6 It can be seen that the fermentation liquid has an inhib...
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