Strain isolation method capable of effectively obtaining effective inoculant of cordyceps militaris
A technology for strain separation and Cordyceps militaris, applied in botany equipment and methods, horticulture, application, etc., can solve problems such as instability of Cordyceps militaris strains, unclear boundaries, and difficulty in distinguishing fertile hyphae from Cordyceps militaris plate colonies , to achieve the effect of less variation, simple operation, and increased number of passages
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Embodiment 1
[0026] 1. Experimental time: April 2012:
[0027] 2. Experimental materials and design:
[0028] 1. Strains: The Cordyceps militaris strain was purchased from the Institute of Microbiology, Chinese Academy of Sciences, No. PRB.1235. Strain A was isolated by traditional methods; strain B was isolated by the following method:
[0029] Use tweezers to hold the fruiting bodies used for tissue isolation, stay in 80°C sterile water for 1.5 seconds, and then quickly transfer to 4°C sterile water in the refrigerator for 2 minutes.
[0030] 2. Culture medium for liquid shake flasks: 300 grams of peeled potatoes (boiled juice), 10 grams of glucose, 2 grams of peptone, 0.5 grams of potassium dihydrogen phosphate, 1 gram of magnesium sulfate, 1000 ml of water, the pH is natural.
[0031] 3. Nutrient solution for fruiting body production: 2% soybean, 1% glucose, 5% milk powder, 0.5% potassium dihydrogen phosphate, 0.05% magnesium sulfate, vitamin B110mg∕1000ml to make a nutrient solution...
Embodiment 2
[0040] The basic operation is the same as in Example 1, except that the bacterial strain B is obtained by processing and separating by the following method:
[0041] Clamp the fruiting bodies used for tissue separation with tweezers, stay in 75% alcohol for 60 seconds, then quickly transfer to sterile water and wash three times.
[0042] After harvesting, the average fresh weight of strain A reached 28.5 g / bottle. After harvesting, the average fresh weight of strain B reached 32.3 g / bottle.
Embodiment 3
[0044] The basic operation is the same as in Example 1, except that the bacterial strain B is obtained by processing and separating by the following method:
[0045] Pour a small amount of 95% alcohol into a Petri dish, light it, use tweezers to hold the fruiting bodies used for tissue separation, and go back and forth on the blue flame for 1-3 times, and then quickly transfer to a refrigerator at 4°C in the refrigerator. Stay in sterile water for 4 minutes.
[0046] After harvesting, the average fresh weight of strain A reached 27.8 g / bottle. After harvesting, the average fresh weight of strain B reached 33.6 g / bottle.
[0047] The above examples show that the operation of the present invention is simple and easy, the separation effect is good, and the effective inoculum part of the plate colony is easy to identify. Not only the stability of the isolated strain is strong, but also the effect of increasing production is achieved compared with the traditional isolation method...
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