A rice endogenous nitrogen-fixing bacterium that improves crop disease resistance and stress resistance and its application
A technology of pathogenic fungi and microbial strains, applied in plant growth regulators, plant growth regulators, methods based on microorganisms, etc., to achieve broad application prospects, good inoculation effect, and strong competitive adaptability
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Embodiment 1
[0049] Example 1. Isolation and identification of rice endogenous nitrogen-fixing bacteria GDSD112
[0050] 1. Isolation of rice endophytic nitrogen-fixing bacteria
[0051] The specific operation for the isolation of rice endogenous nitrogen-fixing bacteria is as follows: take fresh rice plants (collected from Heilongjiang Province, China), first rinse them with tap water, then soak them in 70% ethanol for 1 min, sterilize the surface with 2% sodium hypochlorite for 10 min, and wash them with sterile water. Rinse 3 times. Under aseptic operation conditions, accurately weigh 10.0g of the sample, grind it into a paste in a sterile mortar, transfer and set the volume to 100ml, and continue to dilute to make serial dilution samples, respectively starting from 10 -4 、10 -5 、10 -6 Take 0.1ml of the diluted solution and spread evenly on the above-mentioned CCM medium and nitrogen-free medium plate respectively, culture upside down at 28°C, and after 3-4 days, pick a single colony...
Embodiment 2
[0073] Example 2, Determination of Nitrogenase Activity of Burkholderia sp. GDSD112 CGMCC No.5037
[0074]The Burkholderia sp. (Burkholderia sp.) GDSD112 CGMCC No.5037 that embodiment 1 obtains carries out nitrogenase activity assay, and specific method is as follows: in 15 * 150mm screw-top glass tubes, add 5ml to improve nitrogen fixation culture The base was made into a slant, inoculated with nitrogen-fixing bacteria, and cultured at 28°C. Azotobacter chroococcum (Azotobacter chroococcum) ACCC11103, which is commonly used in the production of microbial fertilizers, was used as the positive control, and the blank slant without inoculation was used as the negative control, with 3 replicates. After culturing for 72 hours, replace the rubber stopper, inject acetylene gas to make the final concentration 10%, seal it with medical adhesive tape, continue culturing for 72 hours, take 100 μl of reaction gas, measure the amount of ethylene produced by gas chromatography, and calculat...
Embodiment 3
[0077] Embodiment 3, Burkholderia sp. (Burkholderia sp.) GDSD112 CGMCC No.5037 antagonizes pathogenic fungus inhibition rate determination
[0078] The Burkholderia sp. GDSD112CGMCC No.5037 obtained in embodiment 1 is carried out to antagonize pathogenic fungus bacteriostatic rate measurement by two-point confrontation method, and concrete operations are as follows: distance center 2cm on PDA plate The crop pathogenic fungi Sclerotinia sclerotiorum (Sclerotinia sclerotiorum) and Burkholderia sp. (Burkholderia sp.) GDSD112 CGMCC No.5037 were inoculated on the two points respectively, and each screening treatment was repeated three times, so that only the pathogenic fungi were inoculated but not inoculated. A plate of Burkholderia sp. GDSD112 CGMCC No. 5037 was used as a control. Incubate at a constant temperature at 28°C, and measure the colony radius r of the pathogenic fungi on the confrontation plate along the direction of the tested Burkholderia sp. GDSD112 CGMCC No.5037 wi...
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