A Banana Endophytic Bacillus Veles and Its Application
A technology of Bacillus and Velez, which is applied in the field of endogenous Bacillus Velez in bananas, can solve the problems of accelerating the resistance of pathogenic microorganisms, etc., and achieve the effects of good development and application prospects, little impact on the ecological environment, and simple cultivation conditions
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Embodiment 1
[0030] The isolation and purification of embodiment 1 Veles bacillus strain BS
[0031] (1) Preparation of PDA medium: first wash and peel the potatoes, then weigh 200g of potatoes and cut them into small pieces, boil them in water (boil for 20-30 minutes, and it can be pierced by a glass rod), and use eight layers of gauze filter. Add 20g of glucose, add water to 1000mL, stir and mix evenly, add 15g of agar, fully heat and dissolve, then dispense into 250mL Erlenmeyer flasks, sterilize with damp heat at 121°C for 20min, and store for later use.
[0032] (2) Preparation of LB medium: Weigh 10g of tryptone, 5g of yeast extract, and 10g of sodium chloride, add water to 1000mL, stir and mix, add 15g of agar, fully heat and dissolve, then pack into 250mL Erlenmeyer flasks, Sterilize with damp heat at 121°C for 20 minutes, and store for future use.
[0033](3) The samples of banana tissue cultured seedlings were collected in Huadu District, Guangzhou City, Guangdong Province. Aft...
Embodiment 2
[0034] The screening of embodiment 2 Veles bacillus strain BS
[0035] (1) Determination of the activity of Bacillus BS by plate confrontation method
[0036] With No. 4 physiological race FOC4 of banana fusarium wilt as target, place the No. 4 physiological race FOC4 bacterium cake of banana fusarium wilt with a diameter of 7mm on the PDA plate medium, dip a little embodiment 1 step (3) with a sterilized toothpick The bacterium liquid of the obtained strain was streaked about 5 cm away from the fungus cake, and cultured upside down in a 28° C. incubator for 4-5 days. A strain with obvious antibacterial effect on the target bacteria was screened out and named BS. The results showed that Bacillus velesii BS had obvious inhibitory effect on Fusarium wilt fungus FOC4 ( figure 1 ). According to the above method, it is effective against Peronophthora litchii, Colletotrichum siamense, Phomopsis longanae, Neofusicoccum parvum and Nalanthamala psidii These pathogenic bacteria were...
Embodiment 3
[0040] The identification of embodiment 3 Veles bacillus strain BS
[0041] Pick a single colony in 3mL LB liquid medium, place it on a 28°C, 180rpm shaker for overnight culture, extract the DNA of the bacteria according to the method of "OMEGAD3350-1 Bacteria DNA Kit", and store it at -20°C for later use. 16S rRNA general primer 27F / 1492R was used for PCR amplification. After passing the test, Sangon Bioengineering (Shanghai) Co., Ltd. was commissioned to perform sequencing. The nucleotide sequence of 16S rRNA is shown in SEQ ID No.1.
[0042] 27F:5'-AGAGTTTGATCCTGGCTCAG-3';
[0043] 1492R: 5'-GGTTACCTTGTTACGACTT-3'.
[0044] The 16S rRNA sequencing results were submitted to NCBI for sequence comparison. The results showed that the strain BS had high homology with Bacillus velezensis K2, B. velezensis BH21, B. velezensis DH82, Bacillus amyloliquefaciens TUL308, B. amyloliquefaciens WH1G and other strains. Both are as high as 99%. Therefore, it is difficult to judge the ta...
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