A stable and efficient artificial inoculation method of Pleurotus eryngii canker pathogens in shelf life
A technology of artificial inoculation and Pleurotus eryngii, applied in the direction of microorganism-based methods, biochemical equipment and methods, bacteria used in food preparation, etc., can solve the problems of instability and low incidence of inoculation, and achieve stable and efficient pathogenicity Determining the effect of artificial inoculation methods
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Embodiment 1
[0027] Example 1: Inoculum Preparation
[0028] Take out the preserved Lactococcus lactis subspecies SLPE1-3 glycerol bacteria from -80°C, use a sterilized toothpick to pick the glycerol bacteria and inoculate them on a sterile MRS agar nutrient plate, and culture them upside down at 25°C for 48 hours; Pick a single colony with a bacterial toothpick, transfer it to MRS liquid medium, and culture it statically at 25°C for 10 to 12 hours until the OD 600 ≈1.0; transfer the bacterial solution to a 50mL sterile centrifuge tube, centrifuge at 4000g and 4°C for 10 minutes, remove the supernatant, and then suspend and centrifuge once with pre-cooled sterile water; The bacteria water was fully suspended, and the density of the bacteria was adjusted to (1.0~5.0)×10 7 CFU / mL is the pathogen inoculum.
Embodiment 2
[0029] Embodiment 2: Inoculation method experiment
[0030] Choose Pleurotus eryngii with uniform size, and use the following three methods for inoculation: Method 1, use a sterilized scalpel to make a cross wound on the middle and lower part of the pleurotus eryngii stalk, the size of the wound is about 2cm×2cm, and the depth of the wound is about 1mm; Method 2, use a sterilized toothpick to make a cross wound on the middle and lower part of the pleurotus eryngii stalk, the size of the wound is about 2cm×2cm, and the depth of the wound is about 1mm; Method 3, use a scalpel to dig the middle and lower part of the pleurotus eryngii 5mm×5mm, 2mm deep wound. The prepared inoculum of Lactococcus lactis subsp. lactis was inoculated on the above three kinds of wounds respectively, and each wound was inoculated with 20 μL of the inoculum. Sterilized water was used as a control. 24 Pleurotus eryngii were inoculated for each treatment, and the inoculated Pleurotus eryngii was arrange...
Embodiment 3
[0031] Embodiment 3: Morbidity rate and disease index survey method
[0032] 1. Investigate the incidence rate and disease index from 8 days to 12 days after vaccination. The calculation method is as follows:
[0033] Incidence rate=(number of pleurotus eryngii) / (total number of inoculated pleurotus eryngii)×100%
[0034] Disease index = ∑ (the number of Pleurotus eryngii at all levels × the relative level value) / (the total number of inoculated Pleurotus eryngii × 9) × 100
[0035] 2. The grading standard of Pleurotus eryngii canker disease during the shelf life:
[0036] Grade 0: no disease;
[0037] Level 1: The lesion area accounts for less than 5% of the mushroom stalk area;
[0038] Grade 3: The lesion area accounts for 5% to 10% of the mushroom stalk area;
[0039] Grade 5: The lesion area accounts for 11% to 25% of the mushroom stalk area;
[0040] Grade 7: The lesion area accounts for 26% to 50% of the mushroom stalk area;
[0041] Grade 9: The lesion area accoun...
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