Biocontrol strain PA-2 (Pseudomonas Aeruginosa) for preventing and curing tobacco angular leaf spot
A technology of tobacco angular leaf spot and PA-2, which is applied in the direction of bacteria, biocides, microorganisms, etc., can solve the problems of non-cultivated varieties resistant to angular leaf spot and the adverse effects on the safety of tobacco leaf export cigarettes, so as to achieve the benefit of non-toxic Effects of pollution production and cost reduction
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Embodiment 1
[0019] Embodiment 1 can control the biocontrol bacteria-Pseudomonas aeruginosa ( Pseudomonas aeruginosa ) Screening of PA-2
[0020] Specific steps are as follows:
[0021] Isolation of bacteria from barnyardgrass: cut about 2 g from the barnyardgrass plant with sterilized scissors, put it in a sterilized mortar, add 1 mL of sterile water to grind it into a homogenate, and serially dilute the homogenate 10 times to 10 - 7 times, will be 10 - 4 to 10 - 7 100 μL of each dilution was drawn and spread on the NA plate, each concentration was repeated three times, and cultured at 28°C for 2-3 days. Pick different types of colonies, streak and purify them on the NA plate, pick a single colony and transfer it to a PDA slant for numbering and storage for later use; it can also be numbered and stored in 40% glycerol and stored in a -80°C refrigerator.
[0022] Preparation of bacterial fermentation broth: pick out the bacterial cultures of Xanthomonas tabacum and isolated from barny...
Embodiment 2
[0035] Example 2 Pseudomonas aeruginosa ( Pseudomonas aeruginosa ) Optimization of fermentation conditions for PA-2 (Accession No. CGMCC No. 5937)
[0036] The bacterial strain is Pseudomonas aeruginosa (Pseudomonas aeruginosa) PA-2 (preservation number CGMCC No. 5937)
[0037] Specific steps are as follows:
[0038]Single factor experiment: NB culture medium was used as the basal medium, and different single carbon sources were selected (glucose, sucrose, lactose, xylose, mannitol maltose, soluble starch and glycerol, with an amount of 0.5%); Select different single organic nitrogen sources (tryptone, peptone, beef extract and yeast extract, dosage 1%), different single inorganic nitrogen sources (NH 4 Cl, (NH 4 ) 2 SO 4 、KNO 3 and NH 4 NO 3 , dosage 1%); in the case of better carbon source and nitrogen source, add different concentrations of NaCl (0.1%, 0.3% and 0.5%); better carbon source, nitrogen source and NaCl, add different concentrations of inorganic nitrogen ...
Embodiment 3
[0042] Example 3 Pseudomonas aeruginosa ( Pseudomonas aeruginosa ) Field efficacy test of PA-2 (Accession No. CGMCC No. 5937) on tobacco angular leaf spot.
[0043] Pseudomonas aeruginosa ( Pseudomonas aeruginosa )PA-2 (Accession No. CGMCC No. 5937)
[0044] There are 15 tobacco plants in each plot, and the plant spacing is 0.9 m×0.5 m. A total of 5 treatments were set up: A biocontrol bacteria PA-2 viable fermentation broth (6-fold dilution); B biocontrol bacteria PA-2 optimized fermentation broth (6-fold dilution); C NB culture medium control (6-fold dilution solution); D optimized NB culture medium control (6-fold dilution); E pharmaceutical streptomycin (3000-fold dilution). Arranged in random blocks and repeated 3 times. Each treatment was sprayed for the first time by high-pressure spraying method at the tree cluster stage, and A, B, C, D, and E were sprayed with each treatment liquid, and each treatment was sprayed with 2 L. Two days later, each treatment was spray...
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