Bacillus pumilus s35 and application thereof
A technology of Bacillus pumilus and strains, applied in the field of microorganisms, can solve the problems of insufficient biological control technology of potato dry rot, and achieve the effects of low cost, good control effect, effective antagonism and control
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Embodiment 1
[0027] Example 1 Isolation and Identification of Bacillus pumilus S35
[0028] 1.1 Isolation and purification of Bacillus pumilus S35
[0029] 1) Water sample pretreatment: Under sterile conditions, the water sample was mixed evenly, filtered with a filter equipped with a filter membrane with a pore size of 0.22 μm, and bacterial cells were enriched on the filter membrane. When the amount of water sample used for enrichment reaches 30ml, remove the filter membrane and put it into a glass test tube filled with 3ml of sterile seawater for 10 -1 watery.
[0030] 2) Soil sample pre-treatment: Take an appropriate amount of soil sample, air-dry (about 20 days), and treat at 120°C for 1 hour. Weigh 10g of the treated soil sample, add 90ml of sterile seawater, put it into a sterilized Erlenmeyer flask equipped with glass balls, shake it fully for 30min, and draw the supernatant after standing still, the supernatant is 10 -1 Soil sample. the above 10 -1 Samples were serially dilut...
Embodiment 2
[0058] Antagonism of Example 2 Bacillus pumilus S35 to pathogenic fungi
[0059] 1.1 Bacteria culture
[0060] Inoculate 4 strains of pathogenic fungi (Qing 9A-4-13, Qing 9A-5-2, Qing 9A-5-10, 65B-2-6) into PDA medium, and culture them in a constant temperature incubator at 28°C for one week; Bacillus pumilus S35 was inoculated in medium E and cultured in a 37°C incubator for 1 day.
[0061] 1.2 Consistent training
[0062] Using the "cross-hatching method", the pathogenic fungus was punched into a bacterium cake with a diameter of 5 mm and placed at a distance of 2.5 cm from the center of Bacillus pumilus (colony diameter of 5 mm), and confronted culture was carried out in a petri dish with a diameter of 9 cm. The results are shown in Table 3.
[0063] Table 3 The confrontation culture between s35 and 4 strains of potato dry rot pathogenic fungi
[0064]
[0065] It can be seen from Table 3 that there was no significant difference between the colony radius and the widt...
Embodiment 3
[0066] Example 3 Re-screening of potatoes in vivo
[0067] 1.1 Preparation of fermentation broth
[0068] The bacteria cultivated in Example 2 were inoculated into an Erlenmeyer flask filled with sterile water, placed in a constant temperature shaker at 37° C. and 200 r / min, and cultured with shaking for 5 days before use.
[0069] 1.2 Preparation of bacterial suspension
[0070] Inoculate the bacteria cultured in Example 2 into a Erlenmeyer flask filled with sterile water, place it in a constant temperature shaker at 37°C and 200 r / min for 2 hours, shake well and set aside.
[0071] 1.3 Test method
[0072] Prick two diameters of 5mm and 3mm deep wounds on the sterilized potato fruit with a puncher with a diameter of 5mm (the potato fruit was soaked in 2% NaClO solution for 10min, rinsed with tap water, and dried naturally). Inoculate 40 μl of pathogenic bacteria suspension first, and then inoculate the same volume of Bacillus pumilus suspension. The control was first ino...
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Abstract
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