Main points of inoculation technology of edible fungi

Inoculation is one of the key links in the production of edible fungi. If the inoculation technology of female oysters does not pass, it will cause miscellaneous bacteria pollution, the survival rate is low and all previous efforts are wasted, not to mention cultivation. Whether it is strain isolation, passage or propagation, it should be carried out in strict accordance with aseptic operating procedures under aseptic conditions. The specific points are as follows:

1 personnel quality

1.1 vaccination personnel should have the spirit of being serious, meticulous, conscious and responsible for the cause.

1.2 vaccination personnel should firmly establish the concept of asepsis, there can be no fluke psychology, must not be careless.

1.3 inoculators have the ability to identify the advantages of strains, and unqualified strains can not be used in production.

2 Environmental disinfection

2.1 for newly made or idle vaccination boxes for more than 15 days, 0.1% mercury water or 2% Lysol should be thoroughly scrubbed and disinfected inside and outside the box before being used for the first time.

2.2 before the inoculation room and buffer room which have been idle for more than 30 days are reused, the walls should be sprayed and sterilized with 2% Lysol. For newly renovated or unused inoculation rooms that have been idle for more than 3 months, the indoor walls should be painted with lime paste and the cement floor should be washed with clean water before use.

3 space disinfection

After putting the medium to be inoculated into the inoculation box or inoculation room, it is necessary to sterilize the space in the box or room.

3.1 if it is an original seed with a mother passage or a small amount of inoculation, the inoculation box can be put into the vaccination tool at the same time, and then the space can be sterilized for 30 minutes with chemical fumigation or ultraviolet radiation.

3.2 if it is a large amount of original or tertiary seed production, the culture medium is put into the inoculation room, at the same time into the inoculation tools, and then fumigated with chemical drugs, at the same time, the space is disinfected with ultraviolet radiation for about 40 minutes.

3.3 work clothes, disinfectant, basin, etc., should be placed in the buffer room. All preparations should be done in the buffer room before vaccination. Before entering, turn on the ultraviolet light for 30 minutes. When you leave the buffer room and enter the inoculation room, you still need to turn on the ultraviolet light until the end of the inoculation.

4 carrier disinfection

4.1 cut off the cotton plug tube or bottle mouth of the mother seed test tube or the original seed bottle (bag), scrub the outer wall with 75% alcoholic cotton, the original seed bottle (bag) wash the outer wall with 0.25% potassium permanganate water, and then bring it into the inoculation box (room) with the inoculators.

4.2 vaccination personnel shall not enter or leave the inoculation room casually after changing clothes and washing hands in the buffer room, so as to avoid air convection as little as possible.

5 sterilization

Use alcohol flame to carry out necessary sterilization.

5.1 first, the inoculation tools (hooks, shovels, knives, etc.) are cauterized on the alcohol lamp flame and placed in a clean empty test tube for cooling.

5.2 place the test tube mother or original bottle on the universal clip, rotate to remove the cotton plug, hold the alcohol lamp in the right hand, let the flame touch the mouth of the tube or bottle, rotate the test tube or original bottle slowly for 3 to 4 weeks with the left hand, and then secure the test tube or original bottle.

6 inoculation

During the separation, passage or propagation of the mother species, the test tube mouth should be controlled within the range of 10cm from the alcohol flame.

6.1 use a sterilized spare inoculation hook to remove the tip of the mother slope and the central mother block, and then put the inoculation hook back into the parent test tube.

6.2 the left hand holds the pre-inoculated test tube culture medium, the tube mouth is tilted slightly downward, and the inoculation hook is held with the right little thumb and ring finger, index finger and middle finger. The seed block about 2mm in diameter is taken from the mother test tube, put into the center of the bevel of the pre-inoculation medium, pull out the inoculation hook, put back into the mother test tube, gently burn the cotton plug held by the right hand, and rotate into the inoculated test tube, and put it into the appropriate position from the right hand. Complete all pre-inoculation test tubes continuously.

Note: ① should not be vaccinated for more than 2 hours at a time. During ② operation, the tip of the inoculation tool and the seed block should not touch the tube mouth and other external objects. If the tip of the tool touches the external object, the tool should be replaced, and the seed block will be invalidated when it touches the external object. The general seed quantity of ③: the mother seed of each test tube can reproduce about 100 first-class species, and the original species can be expanded to 8 bottles.

7 two special inoculation methods

When Armillaria mellea is propagated, it should be carried out when there is condensed water in the culture medium, the diameter of seed block should not be less than 0.5mm, and the fungal cord should be carried out at the same time. Each test tube culture medium can be connected with three mother seeds at the same time, and each mother seed should be pressed until the mother seed is trapped in the culture medium about 2mm, so that the bacterial cord can be extended into the culture medium as soon as possible; when inoculating the original seed, each test tube can be inoculated with 3 bottles of the original seed.

During the propagation of 7.2yuan mushroom, the diameter of seed tuber should not be less than 0.3mm. During inoculation, the seed block was drawn longitudinally from the tail of the test tube culture medium about 1.5cm to the tip of the slope about 2cm. This method can shorten the culture period by about 3 days.

8 safe

8.1 A lighted alcohol lamp should not be near flammable materials such as cotton plugs, plastics, etc.

8.2 the lit alcohol lamp should be put firmly, and the alcohol lamp should be strictly prevented from overturning in the course of operation, so as not to cause fire.

8.3 when the inoculation box or inoculation room uses chemical drugs for space disinfection, the space volume should be calculated and used in accordance with the medication instructions, and the dosage should not be increased arbitrarily to avoid causing drug damage.

8.4 Ultraviolet rays can damage the human body, especially electroophthalmitis. Therefore, do not work with the UV light on, let alone look directly at the UV light that is on.