Why should plant tissue technology be aseptically operated and how to achieve sterility?

I'm a sophomore in high school and I'm doing a plant tissue culture experiment.

Although the callus has not yet been seen (on the third day after vaccination), I am afraid that some of them have been infected with microorganisms.

It's not easy to be sterile, so it's important to experiment a lot (be prepared to fail).

It is important to know that the composition of MS medium is suitable not only for plants, but also for microorganisms. Microorganisms are highly resistant to adversity and are ubiquitous, so it is necessary to reduce the chance of being infected by microorganisms in many ways. The details are as follows:

1.When configuring MS medium, utensils and mother liquor should be autoclaved.

2.Before inoculation, it is necessary to treat different plants and plant organs accordingly. For example:

When I make African violets now, I first have to wash them with detergent, brush off the foreign bodies attached to the plant body with a brush, rinse with water, soak in 75% alcohol solution for 25s, rinse again, soak in 2% sodium hypochlorite solution for 15min, and finally rinse five times before inoculation. (It can also be disinfected with mercuric chloride, calcium hypochlorite, hydrogen peroxide, and antibiotics, but antibiotics will have some plant bodies).

3.Inoculation is performed on an ultra-clean bench, irradiated with a UV lamp for half an hour before use, and the air purification and circulation equipment is turned on.

4.The operator wears a white coat, changes into slippers, and receives a wind shower to blow off the dust before entering the workbench. Use alcohol to disinfect countertops and hands.

Tweezers, blades, etc. during the inoculation process should be burned and sterilized on an alcohol lamp. Of course, the whole process of vaccination should be carried out at the alcohol lamp.

5.Pay attention to air circulation in the culture room.

If aseptic operation is not carried out, then the culture is not a plant but a bacteria.

The culture medium and culture equipment should be sterilized before operation, and aseptic conditions should be maintained during the operation.

Because bacteria can damage tissues, they should be sterilized and disinfected before handling.

Microbial multiplication consumes nutrients and produces toxins, leading to failure in plant tissue culture.

The callus is very fragile, and the presence of foreign bodies can easily affect differentiation.

The culture utensils are aseptically treated, which can be processed at high temperature and on the line.

A certain amount of antibiotics is added to the culture medium to prevent contamination of the culture process.

The culture medium should be replaced regularly in order to remove metabolic waste and prevent the accumulation of metabolites in the cells and cause self-harm to the cells.

Light sterilization is carried out for personnel entering the culture laboratory.

Wear gloves, masks, and white coats ...... when conducting experiments

And the air that comes in should also be sterile.

1. The aseptic operation itself ensures the authenticity of the sample and avoids the contamination of the sample.

2. Aseptic operation also includes the requirements for the operator, so it also protects the safety of the operator itself.

3. Aseptic operation is generally carried out in a specific biological safety cabinet, which also makes use of the characteristics of the biological safety cabinet to ensure that the aerosols formed during the operation will not spread to the environment, so it is also a protection for the environment.

Hello, it is a pleasure to serve you. Due to the plant material used in tissue culture, most of them are composed of young tissues, which are weak in survival after leaving the mother body and are easily infected by bacteria and die. Therefore, in order for these isolated materials to survive and reproduce, it is necessary to prevent bacterial infection and achieve aseptic culture.

Tissue culture media is rich in nutrients, and fungi and bacteria, once multiplied, grow much faster than plant cells, quickly depleting nutrients, leaving plants without nutrients, and at the same time infecting and damaging plants. Therefore, the culture medium and culture utensils must be sterilized. In the process of tissue culture, the sterilization of culture flasks, culture media, utensils, materials, inoculation tools, operating environment, culture room and work surface must be ensured to ensure sterility in order to achieve sterility.

Summary. The role of a sterile operating chamber in plant tissue culture is:

Kiss, look at my **.

1) Establish a sterile system, that is, disinfection and inoculation of explants and culture medium to obtain calli or organs. (2) Proliferation, continuous differentiation to produce new plants, or direct production of adventitious buds and embryoid bodies, can also be repeated as needed to achieve the purpose of mass propagation. (3) The plant is transferred for rooting culture, which can be transferred to the rooting medium, or directly cut for cuttings and rooting.

4) Transition of test-tube seedlings, that is, the adaptation process to the external environment for a certain period of time after the test-tube seedlings are out of the bottle.

Okay, please be a little better, thanks!

Dear, thank you for your patience, after verification: the general steps of aseptic operation of plant tissue culture are as follows: (1) Establish a sterile system, that is, the disinfection and inoculation of explants and culture medium, and obtain callus tissues or organs.

(2) Proliferation, continuous differentiation to produce new plants, or direct production of adventitious buds and embryoid bodies, can also be repeated as needed to achieve the purpose of mass propagation. (3) The plant is transferred for rooting culture, which can be transferred to the rooting medium, or directly cut for cuttings and rooting. 4) Transition of test-tube seedlings, that is, the adaptation process to the external environment for a certain period of time after the test-tube seedlings are out of the bottle.

Dear, do you have any other questions that need to be consulted, if your Bi Han problem has been solved. Dear, due to the epidemic situation, it has caused inconvenience, please understand and tolerate each other, and I believe it will get better soon. Please also pay attention to personal protection.

Okay teacher.

Teacher: If you make a PPT, do you have to prepare it or before the exomorphs?

Teacher: Could you please reply?

Is it something to add after preparation?

Okay, thank you.

For Cong Sheng, it is not easy for you and me to prepare for the pro who will be added after the epidemic. I hope you can give a five-star "I want to evaluate" under the infiltration of the old. I wish you all the best. Peace and quiet. Come on together. Thank you very much.

1. The aseptic operation itself ensures the authenticity of the sample and avoids the contamination of the sample.

2. Aseptic operation also includes the requirements for the operator, so it also protects the safety of the operator itself.

3. Aseptic operation is generally carried out in a specific biological safety cabinet, which also makes use of the characteristics of the biological safety cabinet to ensure that the aerosols formed during the operation will not spread to the environment, so it is also a protection for the environment.

First of all, pay attention to two words: "sterilization" and "sterilization".Obviously, the former is much more demanding than the latter.

As a plant tissue culture, it requires a very strict aseptic environment, and as long as it is stained with a very small amount of bacteria or spores, it may cause culture failure. Moreover, it is not only bacteria that can be contaminated, but also fungi. Therefore, most of the plant tissue culture process is "sterilized" rather than "sterilized".

If the sterilization is not complete, there is contamination in the cultivation process, which will cause the cultivated seedlings to grow slowly or even fail to grow. The principle is as follows: 1) Compete with seedlings for nutrients in the culture medium, and because bacteria and fungi reproduce much faster than plants, the plants do not get nutrients.

2) Parasitic in plants, using raw materials in plant cells to synthesize the fungi's own substances, resulting in the death of seedlings. 3) It has been reported that parasitic bacteria may cause genetic changes within plants, which can lead to the death of seedlings when this gene is necessary for growth. In addition, the genes of the contaminating bacteria can be transferred into the plant cell and expressed through a mechanism whose details are not yet known, leading to culture failure.