Biocontrol strain PNC25 for preventing and controlling litchi downy blight diseases and application of biocontrol strain PNC25
A biocontrol strain, technology of Lychee Downy Phytophthora, applied in the fields of application, bacteria, biocide, etc., can solve the problems that have not yet been seen in the function of Acetyl Microbacterium to prevent and control Lychee Frost Blight, and achieve good development and application prospects, excellent control, and cultivation conditions less demanding effect
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Embodiment 1
[0026] Source of the strain: PNC25 was isolated from the pulp of "Yuan Nuo" in a lychee orchard in Huilai, Jieyang City, Guangdong Province.
[0027] The collected samples were quickly put into numbered plastic bags and brought back to the laboratory for separation. Cut 3 g of pulp, soak in 1 mL of 1% sodium hypochlorite for 5 minutes, soak in 70% alcohol for 2 minutes, and wash with sterile water for 3 times. (Take 0.1mL of the last cleaning solution and apply R2A, or take 0.1mL of the last cleaning solution and add R2A 2 A culture solution, shake culture at 30°C, if it grows aseptically after 48 hours, it is considered that the surface is clean). Grind the surface-sterilized pulp with a sterilized mortar, add 3mL sterile water, filter with sterile cotton cloth, dilute 10 times with sterile water, take 0.1mL to dilute in turn, take 10 -4 、10 -5 、10 -6 Three gradients of 0.1mL coated with R 2 Plate A, cultivated at 28°C for 48 hours, selected plates with 50-300 colonies, ...
Embodiment 2
[0036] Embodiment 2 bacterial agent preparation
[0037] (1) Cultivation of seed bacteria liquid: inoculate PNC25 (GDMCC No.60108) into LB culture liquid, culture at 28°C and 200 rpm for 24 hours, and after 16 hours, take samples every 3 hours in the ultra-clean workbench and measure the OD value at 600nm During the measurement process, use the uninoculated culture solution to adjust to zero, until the OD value of the seed bacteria solution is between 0.5 and 0.8, the culture is ended, and the seed bacteria solution is obtained;
[0038] (2) Preparation of wet bacteria: Add the seed bacteria solution into the LB culture solution at a volume ratio of 1:500, shake and culture at 28°C and 200rpm for 24 hours to obtain a biocontrol agent, and the concentration of viable bacteria is 1×10 8 ~10 9 CFU / mL.
Embodiment 3
[0039] Example 3 Biocontrol bacteria PNC25 evaluates the indoor (LT, laboratory trial) disease prevention effect of litchi fruit / leaf
[0040] The healthy lychees picked in the field are processed as follows: A. Dilute the stock solution of biocontrol bacteria to a final concentration of 5*10 7 CFU / mL, spray fruit / dilute to final concentration 1*10 7 CFU / mL spray leaves; B. LB solution of the same dilution multiple. Set up 3 repetitions for each treatment, each repetition has 30 fruits / 5 branches, and the fruits / branches are placed in a fresh-keeping box (the bottom of the box is covered with sterilized filter paper, 18 cm in diameter, moistened with sterilized water, and the size of the fresh-keeping box is 323*220 *100mm, the manufacturer is Hualong Plastic Factory, Chancheng District, Foshan City). The bacterial solution and the control group were sprayed with a total of 300mL for each treatment (3 repetitions). 24hpt (hours post treatment) spray inoculation of pathogenic ...
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