CN104307007B - Method for flash sterilization of culture medium for tissue culture seedlings factory production - Google Patents
Method for flash sterilization of culture medium for tissue culture seedlings factory production Download PDFInfo
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- CN104307007B CN104307007B CN201410129194.5A CN201410129194A CN104307007B CN 104307007 B CN104307007 B CN 104307007B CN 201410129194 A CN201410129194 A CN 201410129194A CN 104307007 B CN104307007 B CN 104307007B
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- Prior art keywords
- ozone
- culture medium
- outlet valve
- sterilization tank
- air
- Prior art date
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- 230000001954 sterilising effect Effects 0.000 title claims abstract description 42
- 238000004659 sterilization and disinfection Methods 0.000 title claims abstract description 33
- 239000001963 growth medium Substances 0.000 title claims abstract description 21
- 238000000034 method Methods 0.000 title claims abstract description 15
- 238000004519 manufacturing process Methods 0.000 title abstract description 6
- CBENFWSGALASAD-UHFFFAOYSA-N Ozone Chemical compound [O-][O+]=O CBENFWSGALASAD-UHFFFAOYSA-N 0.000 claims abstract description 40
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 21
- 238000010926 purge Methods 0.000 claims abstract description 6
- 239000000463 material Substances 0.000 claims abstract description 5
- 238000000338 in vitro Methods 0.000 claims description 12
- 238000005949 ozonolysis reaction Methods 0.000 claims description 7
- 238000006073 displacement reaction Methods 0.000 claims description 5
- 238000000746 purification Methods 0.000 claims description 5
- 229920003023 plastic Polymers 0.000 claims description 4
- 238000007599 discharging Methods 0.000 claims description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims 1
- 238000004140 cleaning Methods 0.000 claims 1
- 229910052760 oxygen Inorganic materials 0.000 claims 1
- 239000001301 oxygen Substances 0.000 claims 1
- 238000010438 heat treatment Methods 0.000 abstract description 7
- 238000005265 energy consumption Methods 0.000 abstract description 5
- 238000001816 cooling Methods 0.000 abstract 1
- 238000004806 packaging method and process Methods 0.000 abstract 1
- 238000003756 stirring Methods 0.000 description 12
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 8
- 229920000728 polyester Polymers 0.000 description 8
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 6
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 description 6
- BJEPYKJPYRNKOW-REOHCLBHSA-N (S)-malic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O BJEPYKJPYRNKOW-REOHCLBHSA-N 0.000 description 3
- PAWQVTBBRAZDMG-UHFFFAOYSA-N 2-(3-bromo-2-fluorophenyl)acetic acid Chemical compound OC(=O)CC1=CC=CC(Br)=C1F PAWQVTBBRAZDMG-UHFFFAOYSA-N 0.000 description 3
- 229920001817 Agar Polymers 0.000 description 3
- 239000004471 Glycine Substances 0.000 description 3
- SQUHHTBVTRBESD-UHFFFAOYSA-N Hexa-Ac-myo-Inositol Natural products CC(=O)OC1C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C1OC(C)=O SQUHHTBVTRBESD-UHFFFAOYSA-N 0.000 description 3
- 241000234295 Musa Species 0.000 description 3
- 235000018290 Musa x paradisiaca Nutrition 0.000 description 3
- 239000001888 Peptone Substances 0.000 description 3
- 108010080698 Peptones Proteins 0.000 description 3
- 240000000111 Saccharum officinarum Species 0.000 description 3
- 235000007201 Saccharum officinarum Nutrition 0.000 description 3
- 244000061456 Solanum tuberosum Species 0.000 description 3
- 235000002595 Solanum tuberosum Nutrition 0.000 description 3
- 239000008272 agar Substances 0.000 description 3
- BJEPYKJPYRNKOW-UHFFFAOYSA-N alpha-hydroxysuccinic acid Natural products OC(=O)C(O)CC(O)=O BJEPYKJPYRNKOW-UHFFFAOYSA-N 0.000 description 3
- 239000004615 ingredient Substances 0.000 description 3
- CDAISMWEOUEBRE-GPIVLXJGSA-N inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 description 3
- 229960000367 inositol Drugs 0.000 description 3
- 239000001630 malic acid Substances 0.000 description 3
- 235000011090 malic acid Nutrition 0.000 description 3
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 3
- 235000019796 monopotassium phosphate Nutrition 0.000 description 3
- 238000012856 packing Methods 0.000 description 3
- 235000019319 peptone Nutrition 0.000 description 3
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 3
- 239000004323 potassium nitrate Substances 0.000 description 3
- 235000010333 potassium nitrate Nutrition 0.000 description 3
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 description 3
- 239000002002 slurry Substances 0.000 description 3
- 229910052723 transition metal Inorganic materials 0.000 description 3
- 150000003624 transition metals Chemical class 0.000 description 3
- 239000003054 catalyst Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000004064 recycling Methods 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 238000003421 catalytic decomposition reaction Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The invention discloses a method for flash sterilization of a culture medium for tissue culture seedlings factory production. The method comprises the steps of dissolving materials required by the culture medium with water according to proportions; dropwise adding ozone water; sub-packaging; putting sub-packaged culture medium into a sterilization tank F with inlet valves K1-K3 and an outlet valve K4; closing the outlet valve K4; introducing ozone generated by an ozone generator D into the sterilization tank; introducing water vapor generated by an atomizer E into the sterilization tank; controlling an ozone concentration at 0.5-30 ppm, a temperature at 5-70 DEG C, a humidity at 20-95%, a pressure at 0.01-0.1 MPa and a time at 1-40 minutes; opening the outlet valve K4; and introducing purified air from the inlet valve K1 to purge and replace residual ozone in the sterilization tank F to an ozone decomposer G to decompose the residual ozone. The method provided by the invention is thorough in sterilization, shortens time of heating and cooling, and greatly reduces energy consumption.
Description
Technical field
The invention belongs to biological field is and in particular to a kind of method of factorial praluction plantlet in vitro culture medium quick sterilization.
Background technology
Drastically increase reproduction speed and the efficiency of seedling using factorial praluction plantlet in vitro, be widely used to various
Woody, herbal plantation.In industrialized tissue culture seedling incubation, a very crucial step is medium sterilization, this
Directly affect the survival rate of immature seedling, the universal practice is that the culture medium preparing is contained in a kind of special vial in steam
Pressure is 135kpa, and temperature sterilizes at being 121 DEG C, although do so can guarantee that culture medium thoroughly sterilizes, there is also some and lacks
Point.One is high energy consumption, and sterilizing heats up slow, and it is long to terminate rear temperature fall time, affects production efficiency；Two are because adopting high-temperature sterilization,
Container used is general to be easily broken all with the resistant to elevated temperatures container such as glass, is difficult to transport for long-distance, and is unfavorable for recycling.Just
It is due to there being these shortcomings, also cannot make plantlet in vitro production process serialization.Therefore, how reducing energy consumption, overcomes above-mentioned lacking
Point is always the problem that tissue culture engineers make great efforts to solve for many years, and have sets about from management, improves effect by scheduling of production
Rate, but nonetheless it is not really achieved satisfied effect yet.
Content of the invention
The shortcomings and deficiencies existing for prior art, it is an object of the invention to provide one kind can be greatly lowered energy
The method that consumption, group culture container adopt the factorial praluction plantlet in vitro culture medium quick sterilization of recyclable materials.
In order to realize above-mentioned technical assignment, the present invention adopts the following technical scheme that and is achieved:
A kind of method of factorial praluction plantlet in vitro culture medium quick sterilization, specifically follows the steps below:
1) by the material needed for culture medium in proportion use water dissolves, then be added dropwise to total water consumption 5%～15% amount 0.4～
5.0mg/l Ozone Water, constant volume is divided in transparent plastic bottle；
2) plastic bottle containing culture medium is put into and there is sealed door, intake valve k1～k3, the sterilization tank f of air outlet valve k4
In, close sealed door, air outlet valve k4, intake valve k1, k2, k3 pass sequentially through pipeline and ozone generator d, atomizer e, air are net
Gasifying device c is connected, and air outlet valve k4 is connected with ozonolysis equipment g by pipeline；The ozone that ozone generator d is produced is passed through
In sterilization tank, the steam that atomizer e is produced is passed through in sterilization tank, control ozone concentration 0.5～30 ppm, 5～70 DEG C of temperature,
Humidity 20～95%, in 0.01～0.1mpa, the time was at 1～40 minute for pressure；
3) open air outlet valve k4, be passed through the air of purification from intake valve k1, the sky of residual ozone in purging displacement sterilization tank f
Gas；
4) ozone-containing air discharging air outlet valve k4 is passed through ozonolysis equipment g, decomposes remaining ozone；
5) sterilizing can be directly entered subsequent processing after terminating.
The invention provides a kind of method of factorial praluction plantlet in vitro culture medium quick sterilization, through actual tests, no
Only sterilize thoroughly and shorten temperature fall time, decrease the loss of nutrients, most importantly greatly reduce energy consumption, due to
The material of the degradable recycling such as polyester can be used as the container of culture medium using room temperature sterilizing, reduce and produce into
This, improve production efficiency, is very easy to be widely popularized on plantlet in vitro produces, it will promote plantlet in vitro factorial praluction to occur
Revolutionary progress.
Brief description
Fig. 1 is factorial praluction plantlet in vitro culture medium Quick sterilizing device structural representation.
Fig. 2 is culture medium agitating device structure chart.
Below in conjunction with drawings and Examples, explanation is further elaborated to technical scheme.
Specific embodiment
Defer to technique scheme, the present embodiment provides a kind of side of factorial praluction plantlet in vitro culture medium quick sterilization
Method, specifically follows the steps below:
Embodiment 1
1) 1000 liters with stirring main batching kettle a in add 700kg water, 0.4kg ammonium nitrate, 0.5kg potassium nitrate,
0.1kg glycine, 0.05kg malic acid, 0.2kg inositol, 0.5kg peptone, 0.2kg potassium dihydrogen phosphate, heating stirring to 45～
55 DEG C, add 1.0kg activated carbon until completely dissolved.100kg water, 4.0kg agar, 20.0kg sugarcane is added in auxiliary ingredients kettle
Add after sugared heating for dissolving in main batching kettle, main batching kettle is stirring evenly and then adding into 100.0kg water, 40kg banana, 20kg
The slurry that potato is made.It is stirred for uniformly adding 100.0kg Ozone Water (4.0mg/l) afterwards, stirring was lowered the temperature and with 80ml- after 5 minutes
100ml fixed molten, be divided in polyester bottles.
2) good for above-mentioned packing polyester bottles are loaded one there is sealed door, into and out of in the sterilization tank of air valve, close airtight
Door, air outlet valve.The ozone that ozone generator is produced is passed through in sterilization tank, control ozone concentration 5ppm, temperature control at 30 DEG C,
Humidity 45～55%, in 0.01mpa, the time was at 15 minutes for pressure.
3) open air outlet valve, be passed through remaining sky ozoniferous the air purging displacement sterilization tank of purification from air inlet
The ozone-containing air that gas outlet is discharged is passed through ozonolysis equipment simultaneously and decomposes remaining ozone by gas, to be replaced finish after
Enter subsequent processing.
The catalyst that described catalytic decomposition of ozone device is selected is the oxide of transition metal, catalyst be shaped as grain
The particle that footpath is more than 0.5 millimeter.
The oxide of described transition metal can directly be applied, or the oxide for carried transition metal, this mistake
The carrier crossing the oxide of metal is activated carbon, sio2, al2o3 or diatomite.
Embodiment 2
1) 1000 liters with stirring main batching kettle a in add 720kg water, 0.4kg ammonium nitrate, 0.5kg potassium nitrate,
0.1kg glycine, 0.05kg malic acid, 0.2kg inositol, 0.5kg peptone, 0.2kg potassium dihydrogen phosphate, heating stirring to 45～
55 DEG C, add 1.0kg activated carbon until completely dissolved.100kg water, 4.0kg agar, 20.0kg sugarcane is added in auxiliary ingredients kettle
Add after sugared heating for dissolving in main batching kettle, main batching kettle is stirring evenly and then adding into 100.0kg water, 40kg banana, 20kg
The slurry that potato is made.It is stirred for uniformly adding 80.0kg Ozone Water (4.5mg/l) afterwards, stirring was lowered the temperature and with 80ml- after 5 minutes
100ml fixed molten, be divided in polyester bottles.
2) good for above-mentioned packing polyester bottles are loaded one there is sealed door, into and out of in the sterilization tank of air valve, close airtight
Door, air outlet valve.The ozone that ozone generator is produced is passed through in sterilization tank, control ozone concentration 3ppm, temperature control at 30 DEG C,
Humidity 45～55%, in 0.01mpa, the time was at 30 minutes for pressure.
3) open air outlet valve, be passed through remaining sky ozoniferous the air purging displacement sterilization tank of purification from air inlet
The ozone-containing air that gas outlet is discharged is passed through ozonolysis equipment simultaneously and decomposes remaining ozone by gas, to be replaced finish after
Enter subsequent processing.
Embodiment 3
1) 1000 liters with stirring main batching kettle a in add 750kg water, 0.4kg ammonium nitrate, 0.5kg potassium nitrate,
0.1kg glycine, 0.05kg malic acid, 0.2kg inositol, 0.5kg peptone, 0.2kg potassium dihydrogen phosphate, heating stirring to 45～
55 DEG C, add 1.0kg activated carbon until completely dissolved.100kg water, 4.0kg agar, 20.0kg sugarcane is added in auxiliary ingredients kettle
Add after sugared heating for dissolving in main batching kettle, main batching kettle is stirring evenly and then adding into 100.0kg water, 40kg banana, 20kg
The slurry that potato is made.It is stirred for uniformly adding 50.0kg Ozone Water (5.0mg/l) afterwards, stirring was lowered the temperature and with 80ml- after 5 minutes
100ml fixed molten, be divided in polyester bottles.
2) good for above-mentioned packing polyester bottles are loaded one there is sealed door, into and out of in the sterilization tank of air valve, close airtight
Door, air outlet valve.The ozone that ozone generator is produced is passed through in sterilization tank, control ozone concentration 5ppm, temperature control at 30 DEG C,
Humidity 45～55%, in 0.01mpa, the time was at 20 minutes for pressure.
3) open air outlet valve, be passed through remaining sky ozoniferous the air purging displacement sterilization tank of purification from air inlet
The ozone-containing air that gas outlet is discharged is passed through ozonolysis equipment simultaneously and decomposes remaining ozone by gas, to be replaced finish after
Enter subsequent processing.
From the above it can be seen that using the sterilizing methods in embodiment it is only necessary to culture medium is warmed dissolving, sterilizing is entered at room temperature
Row, with traditional whole culture mediums be steam heated to 121 DEG C sterilize methods compared with, the energy consumption of sterilizing substantially reduces,
Reduction amplitude reaches 60%～90%, simultaneously because sterilising temp is low, the polyester bottles of reproducible utilization can be adopted to replace frangible
Vial, and do not need to lower the temperature for a long time, be directly entered next procedure, be conducive to the serialization producing.
Claims (1)
1. a kind of method of factorial praluction plantlet in vitro culture medium quick sterilization is it is characterised in that specifically enter according to following steps
OK:
1) material needed for culture medium is used water dissolves in proportion, then the 0.4～5.0mg/ being added dropwise to total water consumption 5%～15% amount
L Ozone Water, constant volume is divided in transparent plastic bottle；
2) plastic bottle containing culture medium is put into sealed door, the first intake valve (k1), the second intake valve (k2), the 3rd entered
In air valve (k3), the sterilization tank (f) of air outlet valve (k4), close sealed door, air outlet valve (k4), the first intake valve (k1), second enter
Air valve (k2), the 3rd intake valve (k3) pass sequentially through pipeline and ozone generator (d), atomizer (e), air cleaning unit (c)
It is connected, air outlet valve (k4) is connected with ozonolysis equipment (g) by pipeline；The ozone that ozone generator (d) is produced is passed through
In sterilization tank, the steam that atomizer (e) is produced is passed through in sterilization tank, controls ozone concentration 0.5～30 ppm, temperature 5～70
DEG C, humidity 20～95%, in 0.01～0.1mpa, the time was at 1～40 minute for pressure；
3) open air outlet valve (k4), be passed through the air of purification from the first intake valve (k1), remaining smelly in purging displacement sterilization tank (f)
The air of oxygen；
4) ozone-containing air discharging air outlet valve (k4) is passed through ozonolysis equipment (g), decomposes remaining ozone；
5) sterilizing is directly entered subsequent processing after terminating.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410129194.5A CN104307007B (en) | 2014-04-02 | 2014-04-02 | Method for flash sterilization of culture medium for tissue culture seedlings factory production |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410129194.5A CN104307007B (en) | 2014-04-02 | 2014-04-02 | Method for flash sterilization of culture medium for tissue culture seedlings factory production |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104307007A CN104307007A (en) | 2015-01-28 |
| CN104307007B true CN104307007B (en) | 2017-01-18 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201410129194.5A Expired - Fee Related CN104307007B (en) | 2014-04-02 | 2014-04-02 | Method for flash sterilization of culture medium for tissue culture seedlings factory production |
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Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105594341A (en) * | 2016-03-03 | 2016-05-25 | 杨志恒 | Flower seedling cultivation device |
| CN110651667A (en) * | 2019-11-15 | 2020-01-07 | 农业农村部南京农业机械化研究所 | A kind of sterilization equipment and sterilization method for edible mushroom cultivation material |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102668952A (en) * | 2012-04-26 | 2012-09-19 | 中国农业大学 | Sterilization method of soilless culture medium |
| CN102771315A (en) * | 2012-08-17 | 2012-11-14 | 四川宜爱菌业有限公司 | Sterilization method for edible mushroom culture |
| CN103110968A (en) * | 2013-01-25 | 2013-05-22 | 青岛雪洁助剂有限公司 | Energy-conservation sterilizing method for microbial fermentation |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| SG137723A1 (en) * | 2006-05-31 | 2007-12-28 | Aqua Active Singapore Pte Ltd | An ozonised vapour generator |
| US8641985B2 (en) * | 2011-08-25 | 2014-02-04 | Tuttnauer Ltd. | Control system for ozone sterilization in multiple compact chambers |
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2014
- 2014-04-02 CN CN201410129194.5A patent/CN104307007B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102668952A (en) * | 2012-04-26 | 2012-09-19 | 中国农业大学 | Sterilization method of soilless culture medium |
| CN102771315A (en) * | 2012-08-17 | 2012-11-14 | 四川宜爱菌业有限公司 | Sterilization method for edible mushroom culture |
| CN103110968A (en) * | 2013-01-25 | 2013-05-22 | 青岛雪洁助剂有限公司 | Energy-conservation sterilizing method for microbial fermentation |
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