Production and culture of (1) mother seed

① Making mother medium: add 250g potato, 30g agar, 25g glucose and 3g vitamin B to 2000m 1.

Heating in water until agar is completely boiled to obtain mother seed culture medium;

② Mother seed culture: Take 200ml of mother seed culture medium, put it in a 300ml test tube, plug the nozzle with cotton, and sterilize the test tube in an autoclave at 65438 025℃ for 65438 0. 5 hours, then cooled to room temperature, put into a sealed sterile inoculation box, and inoculate 1cmX 1cm mycelium in each test tube.

In step (1), potatoes need to be boiled in water for half an hour in advance, then taken out, boiled in water with agar, glucose and vitamin B until the agar is boiled, then the cooked culture medium is put into a test tube, and the tube mouth is frozen with cotton after solidification. When cultivating the mother seed, before putting the test tube into the inoculation box, it is necessary to wipe the test tube with 75% alcohol and spray the outer wall of the test tube for disinfection.

When putting the test tube into the aseptic inoculation box, it is also necessary to put the lower part for inoculation, such as tray, alcohol lamp, lighter, utensils, inoculation hook and tweezers, into the inoculation box for disinfection. Disinfect in the inoculation box, put the quilt containing the mixture of potassium permanganate and formaldehyde into the inoculation box, and seal the inoculation box during disinfection.

(2) Production and cultivation of original seeds

(1) Making stock culture medium: take 80% wheat, 10% wood needles and 10% wheat bran, cook the wheat, take it out, stir it evenly with wood needles and wheat grains, control the water content to 60-80%, put it into a culture bottle, plug the bottle with cotton, and culture it.

(2) Original seed culture: put the culture bottle and the test tube containing the mother seed into the inoculation box and seal it for half an hour, then inoculate other seeds in the test tube into the culture bottle of the original seed culture medium, and keep the temperature at 20℃.

(3) the production and cultivation of comparative culture

(1) culture medium: take 75% wheat, cook it, take it out, and put it into 15% wood needles.

5% wheat bran and 5% mud are evenly stirred, and the water content is 60% -80%. They are packed in polypropylene bags, sealed and sterilized in a sterilizer. They were stored at 100℃ for 4 hours at normal pressure, then taken out and cooled to room temperature.

(2) Cultivation of cultivated seeds: after the polypropylene bag and the culture bottle of the original seed culture medium are sealed in the inoculation box for half an hour, the original seeds cultivated in the step (2) are inoculated into the polypropylene bag towel, and the cultivated seeds can be obtained by culturing at 20 DEG C for 15-20 days.

Before the above step (1), it is necessary to collect fresh Morchella fruiting bodies and cut them into diameters.

2-3mm bacterial block, after autoclaving, take PDA culture code, put the bacterial block on PDA culture medium, and put the PDA culture medium in an incubator at 20℃ to obtain mycelium after several weeks.