Novel Burkholderia cenocepacia XJYC 2 and application in prevention and control of litchi downy blight disease and litchi anthraenose
A Burkholderia, litchi anthracnose technology, applied in the direction of application, bacteria, fungicides, etc., can solve the problem of drug resistance of pathogenic microorganisms, and achieve good development and application prospects, little impact on the ecological environment, and requirements for cultivation conditions low effect
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Embodiment 1
[0028] Embodiment 1: Isolation, purification and preservation of Burkholderia neocepacia
[0029] (1) Preparation of LB medium: Weigh 10g of tryptone, 5g of yeast extract, 10g of sodium chloride, add 1000mL of water and stir to mix, then add 15g of agar, fully heat and dissolve, then pack into 250mL Erlenmeyer flask, 121℃ Sterilize with moist heat for 20 minutes and save for later use.
[0030] (2) Isolation and purification of Burkholderia neocepacia: The soil samples used to isolate antagonistic bacteria were collected from the rhizosphere soil of healthy fruit trees in the litchi orchard of South China Agricultural University. Weigh 10 g of soil sample, add the soil sample into a Erlenmeyer flask filled with 90 mL of sterile saline (0.9%) under aseptic conditions, place it on a shaker at 28° C. at 180 rpm for 30 min, and prepare a dilution factor of 10. -1 Soil dilution solution, let it stand at room temperature for 10 minutes to separate solid impurities such as sand, sto...
Embodiment 2
[0036] Embodiment 2: The confrontation culture method measures the activity of Burkholderia cenocepacia
[0037] (1) The preparation of LB medium is the same as in Example 1.
[0038] (2) Preparation of PDA medium: Potato (peeled) 200g, cut into pieces, add 1000mL of water, filter with four layers of gauze after boiling for 20min, add 10g of glucose, 15g of agar, stir until fully dissolved, add distilled water to make up to volume 1000mL, divided into 250mL Erlenmeyer flasks, sterilized by moist heat at 121°C for 20min, and stored for later use.
[0039] (3) Preparation of carrot culture medium: take 200g carrots, fully extract the juice with a juicer, filter with 16 layers of gauze, make up the water to 1000mL with the filtrate, add 15g agar, fully heat and dissolve, then pack in conical flasks, After sealing, sterilize with damp heat at 121°C for 20 minutes, and store for future use.
[0040] (4) Preparation of a single colony of Burkholderia cenocepacia: Streak inoculatio...
Embodiment 3
[0045] Embodiment 3: Oxford cup method assays the activity of Burkholderia cenocepacia biological agent
[0046] (1) The preparation of the LB medium is the same as in Example 1; the preparation of the LB liquid medium is the same as that of the LB solid medium except that agar is not added, and the weighed medicament is fully dissolved and then packed in conical flasks ( 100mL culture medium per bottle), stoppered, bandaged, sterilized at 121°C for 20min, cooled and stored for later use.
[0047] (2) The preparation of PDA medium and carrot medium is the same as in Example 2.
[0048] (3) Preparation of Burkholderia cenocepacia biological agent: get the LB culture fluid cone that the single colony of Burkholderia cenocepacia prepared in example 2 step (3) inserts step (1) to prepare The bottle is placed at 30° C. and cultured at a shaker speed of 200 rpm for 24 hours to obtain the biological agent of Burkholderia neocepacia.
[0049] (4) The cultivation of Phytophthora lych...
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