CN116458426A - Energy-saving simple plant tissue culture aseptic operation method - Google Patents
Energy-saving simple plant tissue culture aseptic operation method Download PDFInfo
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- CN116458426A CN116458426A CN202310277923.0A CN202310277923A CN116458426A CN 116458426 A CN116458426 A CN 116458426A CN 202310277923 A CN202310277923 A CN 202310277923A CN 116458426 A CN116458426 A CN 116458426A
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- forceps
- tissue culture
- scalpel
- energy
- plant tissue
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- 238000000034 method Methods 0.000 title claims abstract description 26
- 238000004161 plant tissue culture Methods 0.000 title claims abstract description 19
- 230000001954 sterilising effect Effects 0.000 claims abstract description 29
- 238000004659 sterilization and disinfection Methods 0.000 claims abstract description 21
- 239000001963 growth medium Substances 0.000 claims abstract description 13
- 238000004806 packaging method and process Methods 0.000 claims description 9
- 229920000742 Cotton Polymers 0.000 claims description 5
- 239000011088 parchment paper Substances 0.000 claims description 3
- 229920003023 plastic Polymers 0.000 claims description 3
- 238000007789 sealing Methods 0.000 claims description 3
- 239000004744 fabric Substances 0.000 claims 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 abstract description 13
- 238000005265 energy consumption Methods 0.000 abstract description 4
- 238000010438 heat treatment Methods 0.000 abstract description 3
- 239000007789 gas Substances 0.000 description 8
- 244000005700 microbiome Species 0.000 description 7
- 230000008901 benefit Effects 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 230000000694 effects Effects 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000009629 microbiological culture Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- CBENFWSGALASAD-UHFFFAOYSA-N Ozone Chemical compound [O-][O+]=O CBENFWSGALASAD-UHFFFAOYSA-N 0.000 description 1
- 206010053615 Thermal burn Diseases 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 238000003915 air pollution Methods 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 238000002485 combustion reaction Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000005611 electricity Effects 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 239000000446 fuel Substances 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 230000036512 infertility Effects 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 241000894007 species Species 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
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- Life Sciences & Earth Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The invention provides an energy-saving simple aseptic operation method for plant tissue culture, and belongs to the technical field of tissue culture. According to the number of work tables of a tissue culture plant, required forceps and surgical knives are purchased at one time, the forceps and the surgical knives are sealed and placed into an autoclave for one-time sterilization, and the sterilized forceps and surgical knives are placed in a clean environment for later use. Both the existing alcohol (gas) lamp and the infrared sterilizer consume energy and generate a great amount of heat during the use process. While this heat may offset some of the indoor heating costs in winter, this heat needs to be removed by the air conditioner in summer, thereby again increasing power consumption. In contrast, the present invention has very low energy consumption to sterilize forceps and scalpels in an autoclave together with the culture medium.
Description
Technical Field
The invention relates to the field of tissue culture, in particular to an energy-saving simple plant tissue culture aseptic operation method.
Background
The core technology of plant tissue culture is a sterile manipulation technology, the development of which is based on microbial culture technology. Microbial cultures are typically culture of a single microorganism species, and to culture a single microorganism species, it is desirable to isolate or kill other species of microorganisms. This technique of isolating and killing unwanted microorganisms is called aseptic technique.
The most common way of culturing microorganisms is by tube-in-cotton or flask-in-cotton. Because the space height of the culture dish is small, the culture dish is not suitable for the growth of most plants, and therefore, the early stage of plant tissue culture is basically carried out by a test tube or a triangular flask and a cotton plug. Because the cotton plug is plugged at the pipe orifice or the bottle mouth, the edge of the pipe orifice or the bottle mouth can not be effectively covered and ensured to be clean, the method of burning the pipe orifice or the bottle mouth by an alcohol lamp is adopted in a matched way, and the method is used up to now.
Currently, plant tissue culture plants for commercial production of seedlings have entirely eliminated the use of test tubes and triangular flasks, but instead employed screw-cap cylindrical glass flasks. The bottle cap can well cover the edge of the bottle mouth, effectively avoids the accumulation and proliferation of microorganisms near the bottle mouth, and conventionally burns the bottle mouth to prevent microorganism pollution is not necessary in practice.
In addition to culture devices, another aspect of aseptic manipulation is manipulation tools. The sterile handling tools used in plant tissue culture are forceps and long handled knives (scalpels). To ensure sterility, both tools need to be thoroughly sterilized prior to handling. At present, all tissue culture plants adopt two modes for sterilizing forceps and knives, namely flame burning and infrared sterilizer treatment, and energy is consumed in the sterilization process of the two modes.
Disclosure of Invention
The invention aims to provide an energy-saving simple plant tissue culture aseptic operation method, which solves the technical problems that the conventional plant tissue culture aseptic operation needs frequent sterilization of operation tools, consumes energy and wastes time. By purchasing a large number of operating tools, the operating tools are sterilized while the culture medium is sterilized, so that the problems of high energy consumption and low working efficiency are solved effectively.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:
according to the number of work tables in a tissue culture plant, required forceps and scalpels are purchased at one time, the forceps and the scalpels are sealed and placed into an autoclave for one-time sterilization, the sterilized forceps and the scalpels are placed in a clean environment for standby, and the outer surface of the sealed package is irradiated by ultraviolet rays before use and then used.
Further, the number of forceps and scalpels is determined according to the number of work tables of the tissue culture plant, and when the number of work tables of the plant is a, the number of forceps and scalpels is 80x a.
Further, the sterilizing temperature of the autoclave is 121 ℃ or higher.
Further, the forceps and the scalpel are sealed by being put into transparent plastic bags, so that the sharp forceps and the sharp scalpel are prevented from puncturing the packaging bags, and the packaging bags are protected by being wrapped by two layers of parchment paper.
Further, during sterilization, the packaged forceps, the scalpel and the culture medium are sterilized together or are independently sterilized conventionally, and after sterilization, the packaged forceps, the scalpel and the culture medium are placed in a clean indoor environment for later use.
Further, before use, the sterilized forceps, the scalpel and the tissue culture plant material in the bottle are placed in a workbench to be irradiated with ultraviolet rays for a fixed time, and surface sterilization is performed.
Further, in use, the bag is opened along the flow direction of the filtered air of the workbench, and then placed on the inner side of the workbench, and forceps and a scalpel are taken out for operation.
Due to the adoption of the technical scheme, the invention has the following beneficial effects:
(1) The invention obviously reduces the energy consumption. Both the existing alcohol (gas) lamp and the infrared sterilizer consume energy and generate a great amount of heat during the use process. While this heat may offset some of the indoor heating costs in winter, this heat needs to be removed by the air conditioner in summer, thereby again increasing power consumption. In contrast, the present invention has very low energy consumption to sterilize forceps and scalpels in an autoclave together with the culture medium.
(2) Avoiding accidental industrial injury risk. The alcohol (gas) lamp ignited by liquid (gas) fuel and the high-temperature infrared sterilizer are dangerous to some extent, especially when workers are tired in long-time operation, the workers are easy to scald due to improper operation, and even fire is caused by the toppling of the alcohol lamp. Compared with the existing sterilization mode, the sterilization mode is absolutely safe.
(3) The working environment is improved. The sterile operation workshop is usually a relatively closed space, and special gas and smell are generated when forceps and scalpels with culture medium are treated by using an alcohol (gas) lamp or an infrared sterilizer, and particularly, a great amount of oxygen is consumed and carbon dioxide is generated by continuous alcohol combustion, so that air pollution is caused to the whole sterile operation workshop. Compared with the 2 existing sterilization modes, the sterilization mode of the invention does not generate harmful gas and does not negatively influence the physical health of staff.
(4) The working efficiency is improved. The existing sterilization mode of the alcohol (gas) lamp has the lowest working efficiency, because the sterilization of forceps and a scalpel cannot be completed instantaneously, and the sterilization mode of the infrared sterilizer is also more time-consuming than the mode of the invention which does not need to sterilize the appliance any more because the sterilizer needs to be inserted and extracted and the operation process of cold cutting and alternate use is also needed to be put.
(5) Reduces the production cost and has wide application range. Besides tissue culture plants for mass production, small scientific research laboratories such as schools and research institutes, and teaching laboratories such as middle and primary schools can also be used. Because the burning alcohol lamp is not needed to be placed in the workbench, the laboratory safety of middle and primary schools can be better ensured.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention will be described in further detail by referring to preferred embodiments. It should be noted, however, that many of the details set forth in the description are merely provided to provide a thorough understanding of one or more aspects of the invention, and that these aspects of the invention may be practiced without these specific details.
An energy-saving simple plant tissue culture aseptic operation method comprises the following specific steps:
step 1: depending on the number of tissue culture plant stations, the required forceps and scalpels are purchased at one time (e.g., a total of 8 stations, about 80X8 forceps and 80X8 scalpels are required to be purchased at one time).
Step 2: the method comprises the steps of adopting a transparent plastic bag with proper size (a household fresh-keeping bag with a thick spot is proper) which can be sterilized at high temperature (the sterilization temperature of a culture medium for plant tissue culture is 121 ℃ C., so as to ensure that a packaging bag is not deteriorated, and the packaging bag needs to be more than 125 ℃ C.), packaging and sealing tweezers and a scalpel (two layers of parchment paper can be used for wrapping and protecting in order to avoid the sharp tweezers and the sharp scalpel from puncturing the packaging bag);
step 3: placing into autoclave, sterilizing with culture medium, or sterilizing conventionally. After sterilization, the mixture is placed in a clean indoor environment for standby.
Step 4: before use, the product can be put in a workbench for ultraviolet irradiation (ultraviolet rays generate ozone at the same time and also have sterilization effect) with other articles including tissue culture plant materials in bottles, etc. to sterilize the surfaces.
Step 5: when the filter is used, the bag is opened along the flow direction of the filtered air of the workbench and then is placed on the inner side of the workbench, so that the filter is conveniently taken out for operation.
A quantity of forceps and scalpels are purchased at a time and thoroughly sterilized by sterilizing them with the medium. By applying the mode, not only can energy be greatly saved, the method is simple, but also has obvious economic benefit and obvious advantages of popularization in the tissue culture industry.
The existing method requires placing an alcohol (gas) lamp or an infrared sterilizer special for tissue culture in an ultra-clean workbench; 2-3 forceps and 2-3 surgical knives are used by each operator, and the 2-3 forceps and 2-3 surgical knives are used after being sterilized and cooled alternately at high temperature during use.
The evaluation was made on the basis of the relatively conservative case of only 5 years of operation in a tissue culture plant (the longer the operation time the greater the economic benefit of the application of the invention), and 250 working days per year, 8 hours per day. The 1 workbench consumes about 0.8 yuan of alcohol (the heating negative effect and the cost of the alcohol lamp are not counted) every day in the existing sterilization mode adopting a cauterization device, and the average annual cost is 200 yuan; the method adopts the existing method of sterilizing appliances by using an infrared sterilizer, and adopts the infrared sterilizer to purchase 288 yuan, the power is 150 watts (the lowest selling price and the lowest power of a middle-friend enterprise, namely, a Taobao store, are the same later), the electricity charge per degree is calculated to be 0.7 yuan, and the average annual cost charge is 267.7 yuan (the temperature rise negative effect is not calculated). In the mode of the invention, about 80 forceps and 80 scalpels are purchased for 1 time, and the average annual working cost is 104 yuan (the cost of sterilizing together with the culture medium is not counted) according to 3.5 yuan (a great hardware tool-a Taobao shop, 20 cm gun-shaped forceps) for each forceps and 3 yuan (Ling Lie hardware-a Taobao shop, 3 rd scalpel handle) for each scalpel handle.
Besides tissue culture plants for mass production, small scientific research laboratories such as schools and research institutes, and teaching laboratories such as middle and primary schools can also be used. Because the burning alcohol lamp is not needed to be placed in the workbench, the laboratory safety of middle and primary schools can be better ensured.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Claims (7)
1. An energy-saving simple plant tissue culture aseptic technique is characterized in that: according to the number of workbench of tissue culture factory, the required tweezers and scalpel are purchased once, the tweezers and scalpel are sealed and put into autoclave for one-time sterilization, the sterilized tweezers and scalpel are put into clean environment for use, when in use, the tweezers and scalpel are moved into the workbench, and the sealing is opened for use.
2. The energy-saving simple and convenient aseptic technique for plant tissue culture according to claim 1, which is characterized in that: the number of forceps and scalpels is determined according to the number of work tables of the tissue culture plant, and when the number of work tables is a, the number of forceps and scalpels is 80x a.
3. The energy-saving simple and convenient aseptic technique for plant tissue culture according to claim 1, which is characterized in that: the sterilizing temperature of the autoclave is above 121 ℃.
4. The energy-saving simple and convenient aseptic technique for plant tissue culture according to claim 1, which is characterized in that: the forceps and the scalpel are sealed, and the forceps and the scalpel are filled into a transparent plastic bag, so that the packaging bag is prevented from being pierced by the sharp forceps and the sharp scalpel, and the packaging bag is protected by wrapping the packaging bag with two layers of parchment paper or cotton cloth.
5. The energy-saving simple and convenient aseptic technique for plant tissue culture according to claim 1, which is characterized in that: when in sterilization, the forceps, the scalpel and the culture medium which are packaged in a sealing way are sterilized together, or the forceps, the scalpel and the culture medium are independently sterilized in a conventional way, and the forceps, the culture medium and the culture medium are placed in a clean indoor environment for later use after the sterilization is finished.
6. The energy-saving simple and convenient aseptic technique for plant tissue culture according to claim 1, which is characterized in that: before use, the sealed forceps and the scalpel treated by the autoclave are moved into the workbench, and the surface of the sealed bag is irradiated and disinfected for a certain time by utilizing the ultraviolet lamp in the workbench.
7. The energy-saving simple and convenient aseptic technique for plant tissue culture according to claim 6, which is characterized in that: when in use, the bag is opened along the flow direction of the filtered air of the workbench, and then is placed on the inner side of the workbench, and the forceps and the scalpel are taken out for operation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310277923.0A CN116458426A (en) | 2023-03-20 | 2023-03-20 | Energy-saving simple plant tissue culture aseptic operation method |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202310277923.0A CN116458426A (en) | 2023-03-20 | 2023-03-20 | Energy-saving simple plant tissue culture aseptic operation method |
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| CN116458426A true CN116458426A (en) | 2023-07-21 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN202310277923.0A Pending CN116458426A (en) | 2023-03-20 | 2023-03-20 | Energy-saving simple plant tissue culture aseptic operation method |
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| CN (1) | CN116458426A (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102349445A (en) * | 2011-08-11 | 2012-02-15 | 四川省农业科学院园艺研究所 | Method for producing tissue culture seedlings on assembly line with low cost |
| CN103907531A (en) * | 2013-01-06 | 2014-07-09 | 张征 | Plant tissue culture production process |
-
2023
- 2023-03-20 CN CN202310277923.0A patent/CN116458426A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102349445A (en) * | 2011-08-11 | 2012-02-15 | 四川省农业科学院园艺研究所 | Method for producing tissue culture seedlings on assembly line with low cost |
| CN103907531A (en) * | 2013-01-06 | 2014-07-09 | 张征 | Plant tissue culture production process |
Non-Patent Citations (1)
| Title |
|---|
| 任永波等: "《山西省"两区"开发农业产业化科技指南》", vol. 01, 四川科学技术出版社, pages: 282 - 27 * |
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Application publication date: 20230721 |