CN106577275A - Method of tissue culture for paris polyphylla - Google Patents
Method of tissue culture for paris polyphylla Download PDFInfo
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- CN106577275A CN106577275A CN201610929431.5A CN201610929431A CN106577275A CN 106577275 A CN106577275 A CN 106577275A CN 201610929431 A CN201610929431 A CN 201610929431A CN 106577275 A CN106577275 A CN 106577275A
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- 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
- A01H4/008—Methods for regeneration to complete plants
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Abstract
The invention discloses a method of tissue culture for paris polyphylla, which includes the steps of: 1) treatment on terminal buds; 2) inoculation: preparing operators and devices, sorting seedling clusters, inoculating rooting seedlings, inoculating subculture seedlings, marking the seedlings, and culturing the seedlings. During cultivation, through related measures, such as light source control, temperature control, humidity control, dust and bacterium control, monitoring systems and the like, efficiency and effects of the tissue culture for paris polyphylla are improved, so that the product has stable quality. Through the tissue culture for the paris polyphylla, growth period is reduced and reproduction rate is increased, and adverse influence on growth of the paris polyphylla due to disaster climates is effectively reduced.
Description
Technical field
The present invention relates to the method for tissue culture of plant, and in particular to a kind of method of Rhizoma Paridis tissue culture.
Background technology
Rhizoma Paridis are one of rare Chinese medicines, and the whole world has 24 kinds, and China possesses 19 kinds, wherein again with southwest each province
Species and resource particularly enrich, such as Yunnan, Sichuan, Guizhou etc..Rhizoma Paridis principle active component is steroidal saponin, its aglycon master
To be the diosgenin and pennogenin of different spirostane alcohols, and containing aminoacid, sterone, ecdyson, flavonoid glycoside etc.
Compound.Modern pharmacological research shows that there is Rhizoma Paridis antitumor, hemostasis, relieving cough and asthma, antibacterial, antiviral etc. to act on, and its is good
Curative effect be used for the use of medicament by numerous pharmaceuticals industries, be the important Chinese patent medicines such as YUNNAN BAIYAO, GONGXUENING, pyretic toxicity be clear
Main component.
With the fast development of Chinese Medicine Industry, the demand of Rhizoma Paridis medical material is just incremented by with annual 20% amplitude, and it is wild
Production-goods source is but reduced year by year, causes Rhizoma Paridis price to escalate, and seriously constrains the yield and quality of pharmacy corporation.Currently, weight
Building due to long-term excessively excavation, and lacks the protection to its ecological environment mainly by excavation wild resource, causes wild Rhizoma Paridis to provide
Destructive destruction has been suffered in source, endangered.
Current Rhizoma Paridis are mainly bred by seed and rhizome, because there is embryo development not exclusively in Rhizoma Paridis seed, embryo
Breast is hard, obvious " dual dormancy " characteristic such as plumular axis after-ripening, therefore Rhizoma Paridis seed generally needs to experience two winters after planting
Could Germination And Seedling, be typical " biennial seed ", and in its natural state Rhizoma Paridis seeding ratio is very low, growth cycle
It is long, the needs time of 8～10 years from emerging to becoming a useful person.
The content of the invention
The present invention provides that a kind of growth cycle is short, and breeding potential is high, can effectively reduce Catastrophe climate to plant growing not
The method of the Rhizoma Paridis tissue culture that profit affects.
A kind of method of Rhizoma Paridis tissue culture of the present invention, comprises the following steps：
Step（1）：Terminal bud process
Plant terminal bud part is won using scalpel, is rinsed well terminal bud with clear water, with the mercuric chloride of 1/1000 for preparing
Solution, soaks terminal bud 18-22 minutes, and using sterile fluid water terminal bud 2-4 minutes are rinsed.
Step（2）：Inoculation
1. personnel prepare：Tissue culture personnel need to change into before laboratory slippers, wind drench machine dust removal, change aseptic clothes, wear mask,
Wear and be allowed for access after hair net tissue culture room, handwashing disinfection is needed before operation, speech is forbidden during operation.
2. equipment prepares：Using 75% alcoholic solution sterilization tissue culture table top；Open ultraviolet germicidal 8-12 minutes afterwards；Open
Open high-temperature sterilization device（Tweezers and scalpel are carried out with temperature sterilization to use）, workbench ventilation is opened, prepare long handle tweezers, long handle operation
Knife is each two sets（Convenient sterilizing and rotating of using）, aseptic paper bag is opened, aseptic kraft paper is tiled on the table.
3. Seedling cluster sorting：The right hand holds long handle tweezer, and Seedling cluster is carefully taken out from culture bottle, is placed on aseptic paper.Left hand is held
Scalpel, the Seedling cluster to taking out is sorted by extent of growth, and the seedling for having developed root system is referred to as Seedling of taking root, and not yet develops
The tissue of root system is referred to as Regenerated plant.
4. Seedling of taking root is inoculated with：Take the culture bottle equipped with culture medium, bottle cap is opened on work top, body is edge-on, from
The Seedling of taking root that nip length is similar on aseptic paper, in being planted to culture bottle, notes root being inserted between culture medium, per plant during plantation
Away from relatively uniform, per bottle of plantation 20-25.
5. Regenerated plant inoculation：Same method, the culture bottle equipped with culture medium of taking, opens bottle cap, bottle on work top
Body is edge-on, and the Regenerated plant for growing fine is picked up from aseptic paper, in being planted to culture bottle, bottom touching culture is noted during plantation
Base.
6. labelling：Culture bottle to having planted carries out information flag, comprising plant variety, algebraically, seedlings picking date, operator
Deng.
7. cultivate：The culture bottle being inoculated with, according to internal plant type and the difference of growing way, the training of different condition is put into
Cultivated foster room.
A kind of method of above-mentioned Rhizoma Paridis tissue culture, wherein step（2）Described culture medium its preparation method include with
Lower step：1. mother solution is prepared：By MgSO4·7H2O 280mg、KH2PO4 250 mg、CaCl2 430mg、(NH4)2·
SO4850mg、KCL 2000mg 、H3BO3 3.0 mg、MnSO4·4H2O 22.3 mg、ZnSO4·7H2O 3.8mg、CuSO4·
5H2O 0.025mg、Na2-EDTA37.3mg、FeSO4·7H2O 27.8mg, inositol 55.5mg, the mg of glycine 2.0, Folic Acid
0.50 mg, thiamine hydrochloride 0.25mg, the mg of pyridoxine hydrochloride 1.25, the mg of agar 8000, sucrose 30000mg, 6- benzyl ammonia are fast
The mg of purine 2.0, the mg of indolebutyric acid 1.0, pour in container, adjust pH value 5, it is standby；
2. squeezing juice：Fructus Musae peeling 1.5Kg, Fructus Mali pumilae removal core 500g, using wall-breaking machine blended fruit juice is squeezed into；
3. tentatively mix：Fruit juice slowly being poured into mother solution along chamber wall, being stirred clockwise when pouring into, mixing speed keeps equal
It is even, stirring rod touching container bottom, wall should be avoided in whipping process, in order to avoid produce bubble；
4. dilute：After filter water is boiled, along chamber wall mixed liquor is slowly poured into, stirred clockwise when pouring into, stirred
Speed keeps uniform, and stirring rod touching container bottom, wall should be avoided in whipping process, in order to avoid produce bubble；
5. carbon dust is added：Carbon dust 500mg is uniformly sprinkled into, same method stirs, carbon dust has the effect for expediting the emergence of root system,
If simply expanding the scale of production increasing, without the need for expediting the emergence of root system, then this step is skipped, the culture medium for being not added with carbon dust is translucent breast
White；
6. the subpackage of culture medium：According to the habit of test tube seedling, different types of test tube bottle subpackage, Herba Dendrobii growth bottle are selected
750ml, bottom width mouth is narrow, using drainage screen ventilation bottle cap, flower growth bottle 250ml, the same footpath of base opening, using seal bottle cap；Culture
Base measuring cup carries out subpackage, and bottle 30ml -40ml, big bottle 70-100ml, culture medium is answered uniform speed slow, kept away when pouring culture bottle into
Exempt from culture medium and splash bottleneck, bottle wall.Add a cover after the completion of subpackage, be neatly placed on culture sieve；
7. culture medium sterilization：Carried out disinfection using the high-temperature sterilization device culture medium good to subpackage, in the state of 120 degrees Celsius,
Kept for 40 minutes；
8. culture medium cooling：By sterile channel, the culture medium for disinfecting is proceeded to into cooling chamber, after natural cooling, culture medium by
Liquid is changed into fruit jelly state, and the culture medium shelf-life after cooling is 45 days.
A kind of method of above-mentioned Rhizoma Paridis tissue culture, wherein step（2）Described culture, including following characteristics：
1. in order to provide suitable growth light source, tissue culture room adopts the light control system of natural light+artificially feed, outdoor optical to shine and exceed
During plant demand, light is reduced by the closure of curtain and shutter and is illuminated；When outdoor optical is according to deficiency, using the base of RGB three
The LED light source of color, to tissue culture room light filling is carried out.
2. temperature control：The needs of temperature are not quite similar by the different growth conditions of plant, culturing room employ air-conditioning+
The integrated mode of heater is reaching the control to temperature.
3. humid control：Tissue culture room further provided with humidistat, to ensure the humidity of tissue culture room in rational scope.
4. dirt, bacterium control：In order to ensure that tissue culture room is dustless, aseptic, tissue culture room all personnel gateway adopts double-channel door
Design, and wind be installed between Liang Men drench machine, reduction personnel come in and go out and bring the possibility of bacterium dirt into；The all daylighting windows in tissue culture room are tight
Lattice are sealed, it is to avoid the entrance of winged insect and dust；Tissue culture room conditioning air outlet be equipped with air filter, it is ensured that gout system it is clear
It is clean.
5. monitoring system：In order to improve the environmental index of tissue culture room, every tissue culture room placed alarm, once there is temperature
Degree, humidity, illumination and dust, then can system alarms beyond default warning value.
The present invention compared with prior art, with obvious beneficial effect, as can be known from the above technical solutions:The present invention passes through
Culture to Rhizoma Paridis tissue makes its growth cycle shorten short and improve breeding potential while Catastrophe climate pair can be reduced effectively
The adverse effect of Rhizoma Paridis growth, the present invention is simple to operate, and by tissue culture room light control system, the temperature of natural light+artificially feed are adopted
The enforcement of the related measures such as degree control, humid control, dirt, bacterium control, monitoring system, more improves the effect of Rhizoma Paridis tissue culture
Rate and effect so that the product quality for obtaining is stablized.
Specific embodiment
Embodiment 1：
A kind of method of Rhizoma Paridis tissue culture, comprises the following steps：
Step（1）：Terminal bud process
Plant terminal bud part is won using scalpel, is rinsed well terminal bud with clear water.With the mercuric chloride of 1/1000 for preparing
Solution, immersion terminal bud 22 minutes rinses terminal bud 2 minutes using sterile fluid water.
Step（2）：Inoculation
（1）Personnel prepare：Tissue culture personnel need to change slippers, wind pouring machine dust removal into before laboratory, change aseptic clothes, wear mouth
Cover, wear and be allowed for access after hair net tissue culture room, handwashing disinfection is needed before operation, speech is forbidden during operation.
（2）Equipment prepares：Using 75% alcoholic solution sterilization tissue culture table top；Open ultraviolet germicidal 8 minutes afterwards；Open
High-temperature sterilization device（Tweezers and scalpel are carried out with temperature sterilization to use）, workbench ventilation is opened, prepare long handle tweezers, long handle scalpel
Each two sets（Convenient sterilizing and rotating of using）, aseptic paper bag is opened, aseptic kraft paper is tiled on the table.
（3）Seedling cluster is sorted：The right hand holds long handle tweezer, and Seedling cluster is carefully taken out from culture bottle, is placed on aseptic paper.Left hand is held
Scalpel, the Seedling cluster to taking out is sorted by extent of growth, and the seedling for having developed root system is referred to as Seedling of taking root, and not yet develops
The tissue of root system is referred to as Regenerated plant.
（4）Seedling of taking root is inoculated with：
The preparation of culture medium：
1. mother solution is prepared：By MgSO4·7H2O 280mg、KH2PO4 250 mg、CaCl2 430mg、(NH4)2·SO4850mg、
KCL 2000mg 、H3BO3 3.0 mg、MnSO4·4H2O 22.3 mg、ZnSO4·7H2O 3.8mg、CuSO4·5H2O
0.025mg、Na2-EDTA37.3mg、FeSO4·7H2O 27.8mg, inositol 55.5mg, the mg of glycine 2.0, Folic Acid 0.50
Mg, thiamine hydrochloride 0.25mg, the mg of pyridoxine hydrochloride 1.25, the mg of agar 8000, sucrose 30000mg, 6- benzyl ammonia purine 2.0
Mg, the mg of indolebutyric acid 1.0, pour in container, adjust pH value 5, it is standby；
2. squeezing juice：Fructus Musae peeling 1.5Kg, Fructus Mali pumilae removal core 500g, using wall-breaking machine blended fruit juice is squeezed into；
3. tentatively mix：Fruit juice slowly being poured into mother solution along chamber wall, being stirred clockwise when pouring into, mixing speed keeps equal
It is even, stirring rod touching container bottom, wall should be avoided in whipping process, in order to avoid produce bubble；
4. dilute：After filter water is boiled, along chamber wall mixed liquor is slowly poured into, stirred clockwise when pouring into, stirred
Speed keeps uniform, and stirring rod touching container bottom, wall should be avoided in whipping process, in order to avoid produce bubble；
5. carbon dust is added：Carbon dust 500mg is uniformly sprinkled into, same method stirs, carbon dust has the effect for expediting the emergence of root system,
If simply expanding the scale of production increasing, without the need for expediting the emergence of root system, then this step is skipped, the culture medium for being not added with carbon dust is translucent breast
White；
6. the subpackage of culture medium：According to the habit of test tube seedling, different types of test tube bottle subpackage, Herba Dendrobii growth bottle are selected
750ml, bottom width mouth is narrow, using drainage screen ventilation bottle cap, flower growth bottle 250ml, the same footpath of base opening, using seal bottle cap.Culture
Base measuring cup carries out subpackage, and bottle 30ml -40ml, big bottle 70-100ml, culture medium is answered uniform speed slow, kept away when pouring culture bottle into
Exempt from culture medium and splash bottleneck, bottle wall.Add a cover after the completion of subpackage, be neatly placed on culture sieve；
7. culture medium sterilization：Carried out disinfection using the high-temperature sterilization device culture medium good to subpackage, in the state of 120 degrees Celsius,
Kept for 40 minutes；
8. culture medium cooling：By sterile channel, the culture medium for disinfecting is proceeded to into cooling chamber, after natural cooling, culture medium by
Liquid is changed into fruit jelly state.
Take the culture bottle equipped with culture medium, bottle cap is opened on work top, body is edge-on, and nip is long from aseptic paper
Short similar Seedling of taking root, in being planted to culture bottle, notes for root inserting culture medium during plantation, and every plant of spacing is relatively uniform, often
Bottle plantation 25.
（5）Regenerated plant is inoculated with：Same method, the culture bottle equipped with culture medium of taking, opens bottle cap, bottle on work top
Body is edge-on, and the Regenerated plant for growing fine is picked up from aseptic paper, in being planted to culture bottle, bottom touching culture is noted during plantation
Base.
（6）Labelling：Culture bottle to having planted carries out information flag, comprising plant variety, algebraically, seedlings picking date, operation
Member etc..
（7）Culture：The culture bottle being inoculated with, according to internal plant type and the difference of growing way, different condition is put into
Culturing room is cultivated.
Embodiment 2：
A kind of method of Rhizoma Paridis tissue culture, comprises the following steps：
Step（1）：Terminal bud process：Plant terminal bud part is won using scalpel, is rinsed well terminal bud with clear water.With preparing
1/1000 mercuric chloride solution, immersion terminal bud 20 minutes rinses terminal bud 3 minutes using sterile fluid water.
Step（2）：Inoculation
1. personnel prepare：Tissue culture personnel need to change into before laboratory slippers, wind drench machine dust removal, change aseptic clothes, wear mask,
Wear and be allowed for access after hair net tissue culture room, handwashing disinfection is needed before operation, speech is forbidden during operation.
2. equipment prepares：Using 75% alcoholic solution sterilization tissue culture table top；Open ultraviolet germicidal 10 minutes afterwards；Open
High-temperature sterilization device（Tweezers and scalpel are carried out with temperature sterilization to use）, workbench ventilation is opened, prepare long handle tweezers, long handle scalpel
Each two sets（Convenient sterilizing and rotating of using）, aseptic paper bag is opened, aseptic kraft paper is tiled on the table.
3. Seedling cluster sorting：The right hand holds long handle tweezer, and Seedling cluster is carefully taken out from culture bottle, is placed on aseptic paper.Left hand is held
Scalpel, the Seedling cluster to taking out is sorted by extent of growth, and the seedling for having developed root system is referred to as Seedling of taking root, and not yet develops
The tissue of root system is referred to as Regenerated plant.
4. Seedling of taking root is inoculated with：Take the culture bottle equipped with culture medium, bottle cap is opened on work top, body is edge-on, from
The Seedling of taking root that nip length is similar on aseptic paper, in being planted to culture bottle, notes root being inserted between culture medium, per plant during plantation
Away from relatively uniform, per bottle of plantation 23.
5. Regenerated plant inoculation：Same method, the culture bottle equipped with culture medium of taking, opens bottle cap, bottle on work top
Body is edge-on, and the Regenerated plant for growing fine is picked up from aseptic paper, in being planted to culture bottle, bottom touching culture is noted during plantation
Base.
6. labelling：Culture bottle to having planted carries out information flag, comprising plant variety, algebraically, seedlings picking date, operator
Deng.
7. cultivate：The culture bottle being inoculated with, according to internal plant type and the difference of growing way, the training of different condition is put into
Cultivated foster room.
Embodiment 3：
A kind of method of Rhizoma Paridis tissue culture of the present invention, comprises the following steps：
Step（1）：Terminal bud process：Plant terminal bud part is won using scalpel, is rinsed well terminal bud with clear water.With preparing
1/1000 mercuric chloride solution, immersion terminal bud 18 minutes rinses terminal bud 4 minutes using sterile fluid water.
Step（2）：Inoculation
1. personnel prepare：Tissue culture personnel need to change into before laboratory slippers, wind drench machine dust removal, change aseptic clothes, wear mask,
Wear and be allowed for access after hair net tissue culture room, handwashing disinfection is needed before operation, speech is forbidden during operation.
2. equipment prepares：Using 75% alcoholic solution sterilization tissue culture table top；Open ultraviolet germicidal 12 minutes afterwards；Open
High-temperature sterilization device（Tweezers and scalpel are carried out with temperature sterilization to use）, workbench ventilation is opened, prepare long handle tweezers, long handle scalpel
Each two sets（Convenient sterilizing and rotating of using）, aseptic paper bag is opened, aseptic kraft paper is tiled on the table.
3. Seedling cluster sorting：The right hand holds long handle tweezer, and Seedling cluster is carefully taken out from culture bottle, is placed on aseptic paper.Left hand is held
Scalpel, the Seedling cluster to taking out is sorted by extent of growth, and the seedling for having developed root system is referred to as Seedling of taking root, and not yet develops
The tissue of root system is referred to as Regenerated plant.
4. Seedling of taking root is inoculated with：Take the culture bottle equipped with culture medium, bottle cap is opened on work top, body is edge-on, from
The Seedling of taking root that nip length is similar on aseptic paper, in being planted to culture bottle, notes root being inserted between culture medium, per plant during plantation
Away from relatively uniform, per bottle of plantation 20.
5. Regenerated plant inoculation：Same method, the culture bottle equipped with culture medium of taking, opens bottle cap, bottle on work top
Body is edge-on, and the Regenerated plant for growing fine is picked up from aseptic paper, in being planted to culture bottle, bottom touching culture is noted during plantation
Base.
6. labelling：Culture bottle to having planted carries out information flag, comprising plant variety, algebraically, seedlings picking date, operator
Deng.
7. cultivate：The culture bottle being inoculated with, according to internal plant type and the difference of growing way, the training of different condition is put into
Cultivated foster room.
The above, is only presently preferred embodiments of the present invention, and any pro forma restriction is not made to the present invention, is appointed
What any is simply repaiied without departing from technical solution of the present invention content according to what the technical spirit of the present invention was made to above example
Change, equivalent variations and modification, still fall within the range of technical solution of the present invention.
Claims (3)
1. a kind of method of Rhizoma Paridis tissue culture, comprises the following steps：
Step（1）：Terminal bud process
Plant terminal bud part is won using scalpel, is rinsed well terminal bud with clear water, with the mercuric chloride of 1/1000 for preparing
Solution, soaks terminal bud 18-22 minutes, and using sterile fluid water terminal bud 2-4 minutes are rinsed；
Step（2）：Inoculation
1. personnel prepare：Tissue culture personnel need to change into before laboratory slippers, wind drench machine dust removal, change aseptic clothes, wear mask,
Wear and be allowed for access after hair net tissue culture room, handwashing disinfection is needed before operation, speech is forbidden during operation；
2. equipment prepares：Using 75% alcoholic solution sterilization tissue culture table top；Open ultraviolet germicidal 8-12 minutes afterwards；Open high
Warm disinfector carries out warm sterilization to tweezers and scalpel, opens workbench ventilation, prepares long handle tweezers, long handle scalpel each two sets
Convenient sterilizing and rotating of using, open aseptic paper bag, and aseptic kraft paper is tiled on the table；
3. Seedling cluster sorting：The right hand holds long handle tweezer, and Seedling cluster is carefully taken out from culture bottle, is placed on aseptic paper；Left hand holds operation
Knife, the Seedling cluster to taking out is sorted by extent of growth, and the seedling for having developed root system is referred to as Seedling of taking root, and not yet develops root system
Tissue be referred to as Regenerated plant；
4. Seedling of taking root is inoculated with：Take the culture bottle equipped with culture medium, bottle cap is opened on work top, body is edge-on, from aseptic
The Seedling of taking root that nip length is similar on paper, in being planted to culture bottle, notes for root inserting culture medium, every plant of spacing phase during plantation
To uniform, per bottle of plantation 20-25；
5. Regenerated plant inoculation：Same method, the culture bottle equipped with culture medium of taking opens bottle cap, body side on work top
It is vertical, the Regenerated plant for growing fine is picked up from aseptic paper, in being planted to culture bottle, notice that culture medium is touched in bottom during plantation；
6. labelling：Culture bottle to having planted carries out information flag, comprising plant variety, algebraically, seedlings picking date, operator；
7. cultivate：The culture bottle being inoculated with, according to internal plant type and the difference of growing way, the culturing room of different condition is put into
Cultivated.
2. the method for Rhizoma Paridis tissue culture according to claim 1, it is characterised in that：Wherein step（2）Described culture
The preparation of base is comprised the following steps：
1. mother solution is prepared：By MgSO4·7H2O 280mg、KH2PO4 250 mg、CaCl2 430mg、(NH4)2·SO4850mg、
KCL 2000mg 、H3BO3 3.0 mg、MnSO4·4H2O 22.3 mg、ZnSO4·7H2O 3.8mg、CuSO4·5H2O
0.025mg、Na2-EDTA37.3mg、FeSO4·7H2O 27.8mg, inositol 55.5mg, the mg of glycine 2.0, Folic Acid 0.50
Mg, thiamine hydrochloride 0.25mg, the mg of pyridoxine hydrochloride 1.25, the mg of agar 8000, sucrose 30000mg, 6- benzyl ammonia purine 2.0
Mg, the mg of indolebutyric acid 1.0, pour in container, adjust pH value 5, it is standby；
2. squeezing juice：Fructus Musae peeling 1.5Kg, Fructus Mali pumilae removal core 500g, using wall-breaking machine blended fruit juice is squeezed into；
3. tentatively mix：Fruit juice slowly being poured into mother solution along chamber wall, being stirred clockwise when pouring into, mixing speed keeps equal
It is even, stirring rod touching container bottom, wall should be avoided in whipping process, in order to avoid produce bubble；
4. dilute：After filter water is boiled, along chamber wall mixed liquor is slowly poured into, stirred clockwise when pouring into, stirred
Speed keeps uniform, and stirring rod touching container bottom, wall should be avoided in whipping process, in order to avoid produce bubble；
5. carbon dust is added：Carbon dust 500mg is uniformly sprinkled into, same method stirs, carbon dust has the effect for expediting the emergence of root system,
If simply expanding the scale of production increasing, without the need for expediting the emergence of root system, then this step is skipped, the culture medium for being not added with carbon dust is translucent breast
White；
6. the subpackage of culture medium：According to the habit of test tube seedling, different types of test tube bottle subpackage, Herba Dendrobii growth bottle are selected
750ml, bottom width mouth is narrow, using drainage screen ventilation bottle cap, flower growth bottle 250ml, the same footpath of base opening, using seal bottle cap；Culture
Base measuring cup carries out subpackage, and bottle 30ml -40ml, big bottle 70-100ml, culture medium is answered uniform speed slow, kept away when pouring culture bottle into
Exempt from culture medium and splash bottleneck, bottle wall；Add a cover after the completion of subpackage, be neatly placed on culture sieve；
7. culture medium sterilization：Carried out disinfection using the high-temperature sterilization device culture medium good to subpackage, in the state of 120 degrees Celsius,
Kept for 40 minutes；
8. culture medium cooling：By sterile channel, the culture medium for disinfecting is proceeded to into cooling chamber, after natural cooling, culture medium by
Liquid is changed into fruit jelly state.
3. the method for Rhizoma Paridis tissue culture according to claim 1, it is characterised in that：Step（3）Described culture, including
Following characteristics：
1. in order to provide suitable growth light source, tissue culture room adopts the light control system of natural light+artificially feed, outdoor optical to shine and exceed
During plant demand, light is reduced by the closure of curtain and shutter and is illuminated；When outdoor optical is according to deficiency, using the base of RGB three
The LED light source of color, to tissue culture room light filling is carried out；
2. temperature control：The needs of temperature are not quite similar by the different growth conditions of plant, and culturing room employs air-conditioning+heating
The integrated mode of device is reaching the control to temperature；
3. humid control：Tissue culture room is equipped with humidistat, to ensure the humidity of tissue culture room in rational scope；
4. dirt, bacterium control：In order to ensure that tissue culture room is dustless, aseptic, tissue culture room all personnel gateway setting using double-channel door
Meter, and wind pouring machine is installed between Liang Men, reduction personnel come in and go out and bring the possibility of bacterium dirt into；The all daylighting windows in tissue culture room are strictly close
Envelope, it is to avoid the entrance of winged insect and dust；Tissue culture room conditioning air outlet is equipped with air filter, it is ensured that the cleaning of gout system；
5. monitoring system：In order to improve the environmental index of tissue culture room, every tissue culture room placed alarm, once have temperature,
Humidity, illumination and dust, then can system alarms beyond default warning value.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109156360A (en) * | 2018-10-28 | 2019-01-08 | 江西兼济堂农业开发有限公司 | A kind of method for tissue culture of Paris polyphylla |
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2016
- 2016-10-31 CN CN201610929431.5A patent/CN106577275A/en active Pending
Non-Patent Citations (3)
| Title |
|---|
| 叶方等: ""重楼植物的栽培技术研究进展"", 《中南药学》 * |
| 徐志刚: ""组培微环境与规模化育苗设施环境调控的研究"", 《中国优秀博硕士学位论文全文数据库 (博士) 基础科学辑》 * |
| 陈翠等: ""云南重楼种苗繁育技术"", 《中国现代中药》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109156360A (en) * | 2018-10-28 | 2019-01-08 | 江西兼济堂农业开发有限公司 | A kind of method for tissue culture of Paris polyphylla |
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