Sterilization, inoculation and Culture of Lentinus edodes medium

Sterilization of one strain medium

The sterilization of culture medium is the key to the success or failure of strain production. There are a large number of microorganisms in the unsterilized medium, which often lead to bacteria pollution and loss to production. At present, atmospheric steam sterilization is commonly used by mushroom farmers.

The atmospheric pressure sterilization stove of atmospheric pressure steam sterilization is made of brick. choose an iron pot with a diameter of 110 centimeters, a drum with a height of 100 centimeters and a diameter of 120 centimeters with brick, and smooth it with cement inside and outside.

After loading the culture medium into the sterilizing pot, cover the drum mouth with a thicker plastic film and hemp bag, bind it firmly with rope, then heat it with high heat, and boil the pot in the shortest possible time. When the plastic film is inflated into the shape of steamed bread, it shows that the pot has been boiled, and the sterilization starts at this time and lasts for 8-10 hours to ensure thorough sterilization. Some mushroom farmers are often careless in sterilization, which leads to the failure of seed production due to serious contamination of miscellaneous bacteria. Atmospheric pressure sterilization to achieve "three prevention", first, to prevent cooling midway, halfway have to cease fire, such as the pot can not reach 100 ℃, within the specified time can not achieve the purpose of sterilization. Second, to prevent the pot from drying, add enough water to the pot before sterilization, and in the process of sterilization, if the amount of water in the pot is insufficient, it is necessary to inject water from the water injection mouth in time. Third, it is necessary to prevent the existence of sterilization dead corners, such as uneven fire position at the bottom of the pot, air leakage from only one end of the drum mouth, and so on.

II. Inoculation and cultivation

The sterilized medium can be inoculated, and the inoculation is to transplant the strain to the medium. Operators engaged in vaccination must establish a strict concept of asepsis so that operators and vaccination tools are strictly aseptic. Therefore, it is necessary to strictly sterilize the inoculation tools, thoroughly disinfect the operating environment, disinfect the hands and clothing of the operator, and strictly abide by the aseptic operating rules. Vaccination should be carried out in the inoculation box or in the inoculation room.

When inoculating, clean up the inoculation box or the sundries in the inoculation room and spray 2% Lysol to disinfect it. Put the culture medium, strains and tools to be inoculated on the inoculation box or inoculation room workbench, fumigate the inoculation box and inoculation room with formalin and potassium permanganate for at least half an hour, use 5 grams of potassium permanganate per cubic meter of space, add 10 ml formaldehyde, wipe your hands, mother test tube or original bottle with 75% alcohol cotton before inoculation, and then light the alcohol lamp for operation. After inoculation, turn off the alcohol lamp and move the inoculated bottle or bag to the culture room for culture. In general, a mother seed can be propagated in 4-5 bottles, and a bottle can be reproduced in 40-50 bottles or 15-20 bags.

The culture room should be kept clean and adjust the temperature, humidity, air, light and other environmental conditions. The suitable temperature for strain culture is 20-25 ℃, and the relative humidity of the air is about 60%. During the germicidal period, dark light bacteria are required, and the culture room should always maintain well-ventilated environmental conditions. according to this management method, the bacteria can grow full of bottles or bags in about 40 days. If the bacteria full of bottles (bags) are not used immediately, they should be stored in an environment below 0-10 ℃ and can be stored for 1-3 months.