A strain of Pseudomonas aeruginosa capable of simultaneously controlling tobacco black shank and root black rot and having a growth-promoting effect and its application
A technology of Pseudomonas aeruginosa and Pseudomonas aeruginosa, applied in the field of microorganisms, can solve problems such as pesticide residues, environmental pollution, drug-resistant pathogens, etc., and achieve the effects of reducing the incidence rate, promoting the growth of tobacco seedlings, and having a good inhibitory effect.
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Embodiment 1
[0030] Embodiment 1, the separation of soil microorganism
[0031] Collect tobacco rhizosphere soil in the tobacco field of Zhongzhuang Village, Fenchen Township, Xiangcheng County, Henan Province, weigh 1g of soil, add 10ml of sterile water to fully shake to obtain a soil suspension, and then shake the soil suspension to make appropriate dilution (can be carried out) Doubling dilution), take 0.1ml of the diluted solution and spread it on the KB solid medium containing 8-hydroxyquinoline (0.05g / L), and culture it upside down in a constant temperature incubator at 28°C for 36h until colonies grow out, pick Colonies with different sizes, colors or shapes were separated and purified by streaking on the surface of KB solid medium, and 14 strains were obtained, which were named PA2101, PA210302, PA210401, PA210402, PA2108, PA210901, PA210902, PA2112, PA2113, PA2115, PA211601, PA2117, PA2118, and PA2120.
Embodiment 2
[0032] Embodiment 2, the screening of biocontrol bacteria
[0033] Use the plate confrontation method to screen biocontrol bacteria: inoculate Phytophthora phytophthora and root black rot pathogen Phytophthora rhizoctonia on PDA medium, culture at 25°C and 28°C for 5-10 days, and sterilize The puncher punches the pathogenic bacteria colony into a circular hyphae block with a diameter of 5 mm. Inoculate the pathogenic bacteria block (Monococcus nicotianae and Phytophthora nicotianae) and the isolated bacteria on the PDA medium, the distance between the pathogenic bacteria block and the isolated bacteria is 20mm, and set the pathogenic bacteria block alone as a control. Each treatment was repeated 3 times, and the results of standoff culture were checked after 1 week. Among them, only PA2101 has the highest bacteriostatic rate against the two strains of pathogenic bacteria (see Table 1).
[0034] Table 1 The antibacterial results of 14 strains isolated from the soil against Ph...
Embodiment 3
[0040] Embodiment 3, identification of biocontrol bacteria
[0041]Strain morphology, physiology and biochemistry: strain PA2101 is a Gram-negative bacterium, cultured on KB medium at 28°C for 48 hours, the shape of the colony is yellow and flat, the surface is slightly smooth and nearly round, the center of the circle is slightly convex, and the color is heavier than the surrounding , the edge is irregular, the colony is translucent, surrounded by a yellow-green diffusible fluorescent substance that is visible to the naked eye, and the color of the colony becomes darker and darker after a few days ( figure 2 ). It cannot grow at 4°C, but can grow at 41°C. Oxidase, arginine dihydrolase, gelatin liquefaction and nitrate reduction tests were positive, starch could not be hydrolyzed, and denitrification test, methyl red and VP tests were all negative.
[0042] Sequencing: the bacterial strain PA2101 was sequenced, and the identification result was Pseudomonas aeruginosa.
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