CN105474831B - A kind of bletilla striata naked seed sterilization method - Google Patents
A kind of bletilla striata naked seed sterilization method Download PDFInfo
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- CN105474831B CN105474831B CN201510942248.4A CN201510942248A CN105474831B CN 105474831 B CN105474831 B CN 105474831B CN 201510942248 A CN201510942248 A CN 201510942248A CN 105474831 B CN105474831 B CN 105474831B
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- 238000004659 sterilization and disinfection Methods 0.000 title claims abstract description 44
- 241001313857 Bletilla striata Species 0.000 title claims abstract description 39
- 238000000034 method Methods 0.000 title claims abstract description 36
- 230000001954 sterilising effect Effects 0.000 title claims abstract description 19
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 26
- 239000002775 capsule Substances 0.000 claims abstract description 25
- 239000008223 sterile water Substances 0.000 claims abstract description 25
- 235000013399 edible fruits Nutrition 0.000 claims abstract description 21
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 20
- 239000004088 foaming agent Substances 0.000 claims abstract description 17
- 239000002893 slag Substances 0.000 claims abstract description 13
- 238000002156 mixing Methods 0.000 claims abstract description 12
- 230000000249 desinfective effect Effects 0.000 claims abstract description 10
- 238000004140 cleaning Methods 0.000 claims abstract description 9
- RCTYPNKXASFOBE-UHFFFAOYSA-M chloromercury Chemical compound [Hg]Cl RCTYPNKXASFOBE-UHFFFAOYSA-M 0.000 claims abstract description 8
- 108010016634 Seed Storage Proteins Proteins 0.000 claims abstract description 7
- 239000012459 cleaning agent Substances 0.000 claims abstract description 7
- 229920001213 Polysorbate 20 Polymers 0.000 claims description 17
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 claims description 17
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 claims description 17
- 230000009514 concussion Effects 0.000 claims description 8
- OTYBMLCTZGSZBG-UHFFFAOYSA-L potassium sulfate Chemical compound [K+].[K+].[O-]S([O-])(=O)=O OTYBMLCTZGSZBG-UHFFFAOYSA-L 0.000 claims description 6
- 229910052939 potassium sulfate Inorganic materials 0.000 claims description 6
- 235000011151 potassium sulphates Nutrition 0.000 claims description 6
- FYDKNKUEBJQCCN-UHFFFAOYSA-N lanthanum(3+);trinitrate Chemical compound [La+3].[O-][N+]([O-])=O.[O-][N+]([O-])=O.[O-][N+]([O-])=O FYDKNKUEBJQCCN-UHFFFAOYSA-N 0.000 claims description 5
- 229910001220 stainless steel Inorganic materials 0.000 claims description 5
- 239000010935 stainless steel Substances 0.000 claims description 5
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 claims 1
- 229910052746 lanthanum Inorganic materials 0.000 claims 1
- FZLIPJUXYLNCLC-UHFFFAOYSA-N lanthanum atom Chemical compound [La] FZLIPJUXYLNCLC-UHFFFAOYSA-N 0.000 claims 1
- 229910017604 nitric acid Inorganic materials 0.000 claims 1
- 230000018044 dehydration Effects 0.000 abstract description 8
- 238000006297 dehydration reaction Methods 0.000 abstract description 8
- 230000006378 damage Effects 0.000 abstract description 5
- 230000004083 survival effect Effects 0.000 abstract description 2
- 230000035784 germination Effects 0.000 description 9
- 229960002523 mercuric chloride Drugs 0.000 description 8
- LWJROJCJINYWOX-UHFFFAOYSA-L mercury dichloride Chemical compound Cl[Hg]Cl LWJROJCJINYWOX-UHFFFAOYSA-L 0.000 description 8
- 241000196324 Embryophyta Species 0.000 description 4
- 239000003814 drug Substances 0.000 description 4
- 238000001035 drying Methods 0.000 description 4
- 230000008595 infiltration Effects 0.000 description 4
- 238000001764 infiltration Methods 0.000 description 4
- 238000011081 inoculation Methods 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 238000011012 sanitization Methods 0.000 description 4
- 238000009288 screen filtration Methods 0.000 description 4
- 238000005406 washing Methods 0.000 description 4
- 238000005336 cracking Methods 0.000 description 3
- 238000005187 foaming Methods 0.000 description 3
- LUEWUZLMQUOBSB-FSKGGBMCSA-N (2s,3s,4s,5s,6r)-2-[(2r,3s,4r,5r,6s)-6-[(2r,3s,4r,5s,6s)-4,5-dihydroxy-2-(hydroxymethyl)-6-[(2r,4r,5s,6r)-4,5,6-trihydroxy-2-(hydroxymethyl)oxan-3-yl]oxyoxan-3-yl]oxy-4,5-dihydroxy-2-(hydroxymethyl)oxan-3-yl]oxy-6-(hydroxymethyl)oxane-3,4,5-triol Chemical compound O[C@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@@H](O[C@@H]2[C@H](O[C@@H](OC3[C@H](O[C@@H](O)[C@@H](O)[C@H]3O)CO)[C@@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O LUEWUZLMQUOBSB-FSKGGBMCSA-N 0.000 description 2
- 241001523383 Achnatherum Species 0.000 description 2
- 241001313855 Bletilla Species 0.000 description 2
- 229920002581 Glucomannan Polymers 0.000 description 2
- 208000032843 Hemorrhage Diseases 0.000 description 2
- 230000003796 beauty Effects 0.000 description 2
- 230000000740 bleeding effect Effects 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 229940046240 glucomannan Drugs 0.000 description 2
- 238000005469 granulation Methods 0.000 description 2
- 230000003179 granulation Effects 0.000 description 2
- 238000007781 pre-processing Methods 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- 241000416162 Astragalus gummifer Species 0.000 description 1
- 206010014080 Ecchymosis Diseases 0.000 description 1
- 229920000084 Gum arabic Polymers 0.000 description 1
- 208000000616 Hemoptysis Diseases 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 229910000831 Steel Inorganic materials 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- 239000005864 Sulphur Substances 0.000 description 1
- 229920001615 Tragacanth Polymers 0.000 description 1
- 206010061577 Ulcer haemorrhage Diseases 0.000 description 1
- 206010051373 Wound haemorrhage Diseases 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
- 230000001476 alcoholic effect Effects 0.000 description 1
- 238000009412 basement excavation Methods 0.000 description 1
- 201000009267 bronchiectasis Diseases 0.000 description 1
- 238000005660 chlorination reaction Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 208000001780 epistaxis Diseases 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000010413 gardening Methods 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 208000035861 hematochezia Diseases 0.000 description 1
- 230000023597 hemostasis Effects 0.000 description 1
- XQBXQQNSKADUDV-UHFFFAOYSA-N lanthanum;nitric acid Chemical compound [La].O[N+]([O-])=O XQBXQQNSKADUDV-UHFFFAOYSA-N 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- QSHDDOUJBYECFT-UHFFFAOYSA-N mercury Chemical compound [Hg] QSHDDOUJBYECFT-UHFFFAOYSA-N 0.000 description 1
- 229910052753 mercury Inorganic materials 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 208000008128 pulmonary tuberculosis Diseases 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000010959 steel Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- RTKIYNMVFMVABJ-UHFFFAOYSA-L thimerosal Chemical compound [Na+].CC[Hg]SC1=CC=CC=C1C([O-])=O RTKIYNMVFMVABJ-UHFFFAOYSA-L 0.000 description 1
- 229940033663 thimerosal Drugs 0.000 description 1
- 235000010487 tragacanth Nutrition 0.000 description 1
- 239000000196 tragacanth Substances 0.000 description 1
- 229940116362 tragacanth Drugs 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
- A01C1/08—Immunising seed
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Soil Sciences (AREA)
- Environmental Sciences (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Pretreatment Of Seeds And Plants (AREA)
Abstract
The invention discloses a kind of bletilla striata naked seed sterilization method, including seed is collected, Fruit pod is removed after bletilla striata capsule is dried and takes out seed, Fruit pod disintegrating slag is filtered out, obtains seed；Seed disinfection, adds in the seed being collected into 75% alcohol under clean environment and foaming agent slightly shakes mixing and completes pretreatment, then using sterile water wash seed；0.1% mercury chloride is added after cleaning and foaming agent slightly shakes mixing and carried out disinfection processing；Clean using sterile water wash after disinfecting, be sterilized seed；Seed storage.The present invention be applied to occur it is damaged, damage by worms or bletilla striata naked seed that dehydration is dry and cracked, after bletilla striata naked seed is handled, the large area Browning occurred during tissue culture can be avoided, the phenomenon for causing tissue culture survival rate seriously to reduce, the capsule range of choice for the bletilla striata tissue culture for being further is more extensive, restrictive condition reduces so that is greatly increased suitable for the capsule amount of tissue culture.
Description
Technical field
The present invention relates to field of plant growing technology, and in particular to a kind of bletilla striata naked seed sterilization method.
Background technology
The rare crude drugs bletilla striata (Bletilla Striata (Thunb.) Reichb.f.) is that the bletilla striata belongs to perennial
Herbaceous plant, it is national key protected wild plants, there is high economic value.The bletilla striata is used as medicine with dry tuber, is had and is received
Hold back hemostasis, the effect of detumescence and promoting granulation, be widely used in the treatment disease such as pulmonary tuberculosis and bronchiectasis hemoptysis, ulcer bleeding.The bletilla striata
In addition to for medicine aspect, bletilla striata stem tuber is rich in Bletilla glucomannan, and Bletilla glucomannan can replace tragacanth rubber powder, gummi arabicum pulveratum to make
It is used to field, the tools such as food, beauty and chemical industry for emulsifying agent, suspending agent have been widely used.
The bletilla striata is distributed widely in China Yangtze river basin each province, and main product is in Guizhou, Sichuan, Hunan, Hubei, Anhui, Henan, Zhejiang
The ground such as river, Shaanxi.It is mostly wild, also there is a small amount of cultivation, harvest within 3~4 years.
The bletilla striata is used as medicine with the string root stem tuber of subterranean root.Function astringing to arrest bleeding, detumescence and promoting granulation, treatment spitting of blood, spit blood, wound
Bleeding, have blood in stool, nosebleed, especially have special efficacy to empsyxis, UGB and traumatism and bleeding.It can also make herbal cuisine, have
Beauty ecchymose removing, the function of fat-reducing.Because the bletilla striata blooms very gorgeous, it potted plant can view and admire again, there is higher ornamental value.Its reason
It is to be influenceed in recent years by medicinal demand and gardening growth in strong demand, China's bletilla striata market demand also showed increased；Excessively
Excavation and habitat destruction cause wild resource amount drastically to decline.Interplanted with woody medicinal species, the unit that can improve soil utilizes
Efficiency.
Bletilla striata happiness is shady and cool, moistening and warm environment, can vegetative propagation, can also carry out generative propagation, because seed is tiny,
Sprout relatively difficult under natural conditions, domestication's seedling largely expands numerous typically by the way of tissue-culturing rapid propagation.The tissue-culturing rapid propagation bletilla striata
Seedling all uses fresh uncracked ripe capsule, and capsule is cut after surface sterilization is carried out and sows seed on culture medium.Should
Kind of method requires the complete free from insect pests of capsule without cracking, and unreasonable tissue culture process pollution in wide area may be caused if selecting.Therefore choose
Substantial amounts of unqualified capsule can be given up during choosing, cause the waste of germ plasm resource.And to bear capsule not too much more for the annual bletilla striata,
Wherein there is considerable part cracking or damaged by worms and be not used to tissue culture, often influenceed because of qualified capsule lazy weight white
Splendid achnatherum tissue culture industry is fast-developing.
The content of the invention
, can be to there is breakage, damaging by worms using this method it is an object of the invention to provide a kind of bletilla striata naked seed sterilization method
Or the dry and cracked naked seed of dehydration is handled, the naked seed after processing is ensured that sterilization is thorough, can be improved with respect to before processing
Ensure germination rate.
For the above-mentioned purpose, a kind of bletilla striata naked seed sterilization method is provided in one embodiment of the present of invention, including：
（1）Collect seed：Fruit pod is removed after bletilla striata capsule is dried and takes out seed, Fruit pod disintegrating slag is filtered out, obtains seed；
（2）Seed disinfection：75% alcohol is added in the seed being collected into and foaming agent slightly shakes and mixed under clean environment
It is even to complete pretreatment, then using sterile water wash seed；0.1% mercury chloride is added after cleaning and foaming agent slightly shakes
Mixing is swung to carry out disinfection processing；Clean using sterile water wash after disinfecting, be sterilized seed；
（3）Seed storage：Disinfection seed is split in clean environment and air-dried to seed into powdered.
The optimal enforcement example of the present invention, in sub-processes are collected, Fruit pod disintegrating slag is screened out using 10 mesh stainless (steel) wires.
The optimal enforcement example of the present invention, foaming agent is Tween-20.
The optimal enforcement example of the present invention, during seed disinfection, the concussion time added after alcohol is 60 seconds；Add chlorination
Time of disinfecting after mercury is 3min ~ 10min.
The optimal enforcement example of the present invention, uses sterile water wash 5 times ~ 8 times after disinfecting；Sterilized water is used after pretreatment
Cleaning 3 times ~ 5 times；Disinfection seed is in powdered and without caking using clean wind 3h ~ 5h to seed in clean environment.
In summary, the present invention has advantages below：
The present invention be applied to occur it is damaged, damage by worms or bletilla striata naked seed that dehydration is dry and cracked, can after bletilla striata naked seed is handled
To avoid the large area Browning occurred during tissue culture, the phenomenon for causing tissue culture survival rate seriously to reduce, further yes
The capsule range of choice of bletilla striata tissue culture is more extensive, restrictive condition reduce so that suitable for tissue culture capsule amount significantly
Increase.
The present invention is by pre-processing and disinfecting, it is ensured that though occur it is damaged, damage by worms or that dehydration is dry and cracked etc. is white
The germination rate of splendid achnatherum naked seed, it can apply to industrialize nursery.
The capsule to be ftractureed by long-term preserve can be used to carry out tissue culture for the present invention, and having broken bletilla striata tissue culture can only
The convention sowed in the fall；Bletilla striata nursery at this stage can be overcome all to be sowed using the sterilization of fresh capsule simultaneously, after preserving for a long time
Capsule can natural cracking, internal seeds are but unable to thorough disinfection when causing the capsule to sterilize, and occurrence of large-area is dirty when causing nursery
The defects of dye.
Embodiment
Embodiment 1
A kind of bletilla striata naked seed sterilization method, comprises the following steps：
（1）Collect seed：Selection occurs damaged and damaged by worms, or because storing the improper capsule for causing dehydration dry and cracked；By the bletilla striata
Capsule removes Fruit pod after drying and takes out seed, and seed mix is used to the stainless steel screen filtration Fruit pod disintegrating slag of 10 mesh, filtered out
Fruit pod disintegrating slag, obtains seed；
（2）Seed disinfection：The alcohol of addition 75% and foaming agent Tween-20 are light in the seed being collected under clean environment
Microseism swings mixing and completes pretreatment, then using sterile water wash seed；0.1% mercury chloride and foaming is added after cleaning
Agent is slightly shaken mixing and carried out disinfection processing；Clean using sterile water wash after disinfecting, be sterilized seed.
Specifically, seed can be added in 100ml syringe, grain weight is no more than 50ml.75% alcohol is sucked,
A drop Tween-20 is instilled, and is gently shaken 60 seconds, patting syringe mixes alcohol and Tween-20 so that seed is immersed in liquid
Body or infiltration.Then alcohol is bled off, sucks sterile water washing 3 times.After cleaning add one drop Tween-20 foaming agent and
40mL0.1% mercuric chloride solutions simultaneously gently concussion sterilization 5min, then ibid method removes mercuric chloride solution and with sterile water wash 6 times.
Above procedure must the sterile working in superclean bench
（3）Seed storage：Disinfection seed is split in clean environment and air-dried to seed into powdered.
Specifically operate and be：The seed of sanitized is drawn out with spoon and is positioned over the inoculation dish for being lined with multilayer aseptic filter paper
In, share out and blow 3h ~ 5h with superclean bench wind-force until seed forms block into powdered without companion, you can uniformly broadcast seed
It is spread in Plates for germination media；Above procedure must the sterile working in superclean bench.
Embodiment 2
A kind of bletilla striata naked seed sterilization method, comprises the following steps：
（1）Collect seed：Selection occurs damaged and damaged by worms, or because storing the improper capsule for causing dehydration dry and cracked；By the bletilla striata
Capsule removes Fruit pod after drying and takes out seed, and seed mix is used to the stainless steel screen filtration Fruit pod disintegrating slag of 10 mesh, filtered out
Fruit pod disintegrating slag, obtains seed；
（2）Seed disinfection：Under clean environment in the seed being collected into add 75% alcohol, foaming agent Tween-20, sulphur
Sour potassium and lanthanum nitrate, the final concentration of wherein potassium sulfate and lanthanum nitrate is respectively 1% and 0.05%；Slight concussion, which mixes, completes pre- place
Reason, then using sterile water wash seed；0.1% mercury chloride is added after cleaning and foaming agent slightly shakes mixing and carried out
Disinfect；Clean using sterile water wash after disinfecting, be sterilized seed.The present embodiment is in preprocessing process
After adding potassium sulfate and lanthanum nitrate, seed can be pre-processed, compared with Example 1, after the naked seed processing of the present embodiment
Germination rate compared to embodiment 1 improves 12% or so；Seed can be reduced to be damaged by thimerosal, improve germination rate, so as to
The phenomenon of germination rate reduction will not occur because naked fruit is damage.
Specifically, seed can be added in 100ml syringe, grain weight is no more than 50ml.Suck 75% alcohol,
Potassium sulfate and lanthanum nitrate, a drop Tween-20 is instilled, and gently shaken 60 seconds, patting syringe makes alcohol, potassium sulfate, nitric acid
Lanthanum and Tween-20 mixing so that seed is immersed in liquid or infiltration.Then alcoholic solution is bled off, sucks sterile water washing 3
It is secondary.A drop Tween-20 foaming agent and 40mL0.1% mercuric chloride solutions and gently concussion sterilization 5min are added after cleaning, then same top
Method removes mercuric chloride solution and with sterile water wash 6 times.Above procedure must the sterile working in superclean bench
（3）Seed storage：Disinfection seed is split in clean environment and air-dried to seed into powdered.
Specifically operate and be：The seed of sanitized is drawn out with spoon and is positioned over the inoculation dish for being lined with multilayer aseptic filter paper
In, share out and blow 3h ~ 5h with superclean bench wind-force until seed forms block into powdered without companion, you can uniformly broadcast seed
It is spread in Plates for germination media；Above procedure must the sterile working in superclean bench.
Embodiment 3
A kind of bletilla striata naked seed sterilization method, comprises the following steps：
（1）Collect seed：Selection occurs damaged and damaged by worms, or because storing the improper capsule for causing dehydration dry and cracked；By the bletilla striata
Capsule removes Fruit pod after drying and takes out seed, and seed mix is used to the stainless steel screen filtration Fruit pod disintegrating slag of 10 mesh, filtered out
Fruit pod disintegrating slag, obtains seed；
（2）Seed disinfection：The alcohol of addition 75% and foaming agent Tween-20 are light in the seed being collected under clean environment
Microseism swings mixing and completes pretreatment, then using sterile water wash seed；0.1% mercury chloride and foaming is added after cleaning
Agent is slightly shaken mixing and carried out disinfection processing；Clean using sterile water wash after disinfecting, be sterilized seed.
Specifically, seed can be added in 100ml syringe, grain weight is no more than 50ml.75% alcohol is sucked,
A drop Tween-20 is instilled, and is gently shaken 80 seconds, patting syringe mixes alcohol and Tween-20 so that seed is immersed in liquid
Body or infiltration.Then alcohol is bled off, sucks sterile water washing 3 times.After cleaning add one drop Tween-20 foaming agent and
60mL0.1% mercuric chloride solutions simultaneously gently concussion sterilization 8min, then ibid method removes mercuric chloride solution and with sterile water wash 7
It is secondary.Above procedure must the sterile working in superclean bench.
（3）Seed storage：Disinfection seed is split in clean environment and air-dried to seed into powdered.
Specifically operate and be：The seed of sanitized is drawn out with spoon and is positioned over the inoculation dish for being lined with multilayer aseptic filter paper
In, share out and blow 3h ~ 5h with superclean bench wind-force until seed forms block into powdered without companion, you can uniformly broadcast seed
It is spread in Plates for germination media；Above procedure must the sterile working in superclean bench.
Embodiment 4
A kind of bletilla striata naked seed sterilization method, comprises the following steps：
（1）Collect seed：Selection occurs damaged and damaged by worms, or because storing the improper capsule for causing dehydration dry and cracked；By the bletilla striata
Capsule removes Fruit pod after drying and takes out seed, and seed mix is used to the stainless steel screen filtration Fruit pod disintegrating slag of 10 mesh, filtered out
Fruit pod disintegrating slag, obtains seed；
（2）Seed disinfection：The alcohol of addition 75% and foaming agent Tween-80 are light in the seed being collected under clean environment
Microseism swings mixing and completes pretreatment, then using sterile water wash seed；0.2% mercury chloride and foaming is added after cleaning
Agent is slightly shaken mixing and carried out disinfection processing；Clean using sterile water wash after disinfecting, be sterilized seed.
Specifically, seed can be added in 100ml syringe, grain weight is no more than 50ml.75% alcohol is sucked,
A drop Tween-20 is instilled, and is gently shaken 90 seconds, patting syringe mixes alcohol and Tween-20 so that seed is immersed in liquid
Body or infiltration.Then alcohol is bled off, sucks sterile water washing 4 times.After cleaning add one drop Tween-20 foaming agent and
60mL0.1% mercuric chloride solutions simultaneously gently concussion sterilization 7min, then ibid method removes mercuric chloride solution and with sterile water wash 7 times.
Above procedure must the sterile working in superclean bench
（3）Seed storage：Disinfection seed is split in clean environment and air-dried to seed into powdered.
Specifically operate and be：The seed of sanitized is drawn out with spoon and is positioned over the inoculation dish for being lined with multilayer aseptic filter paper
In, share out and blow 5h with superclean bench wind-force until seed forms block into powdered without companion, you can uniformly sow seed
In Plates for germination media；Above procedure must the sterile working in superclean bench.
Claims (8)
1. a kind of bletilla striata naked seed sterilization method, including：
(1) seed is collected：Fruit pod is removed after bletilla striata capsule is dried and takes out seed, Fruit pod disintegrating slag is filtered out, obtains seed；
(2) seed disinfection：Under clean environment in the seed being collected into add 75% alcohol and foaming agent, potassium sulfate and nitric acid
The final concentration of lanthanum, wherein potassium sulfate and lanthanum nitrate is respectively 1% and 0.05%；Slight concussion, which mixes, completes pretreatment, then makes
With sterile water wash seed；0.1% mercury chloride is added after cleaning and foaming agent slightly shakes mixing and carried out disinfection processing；
Clean using sterile water wash after disinfecting, be sterilized seed；
(3) Seed storage：Disinfection seed is split in clean environment and air-dried to seed into powdered.
2. the method as described in claim 1, it is characterised in that：In the collection sub-processes, 10 mesh stainless steel mesh screens are used
Except Fruit pod disintegrating slag.
3. the method as described in claim 1, it is characterised in that：The foaming agent is Tween-20.
4. the method as described in claim 1, it is characterised in that：During the seed disinfection, when adding the concussion after alcohol
Between be 60 seconds.
5. the method as described in claim 1, it is characterised in that：During the seed disinfection, the sterilization after addition mercury chloride
Processing time is 3min~10min.
6. the method as described in claim 1, it is characterised in that：It is described disinfect after use sterile water wash 5 times~8 times.
7. the method as described in claim 1, it is characterised in that：Sterile water wash is used after the pretreatment 3 times~5 times.
8. the method as described in claim 1, it is characterised in that：The disinfection seed uses clean wind 3h in clean environment
~5h is in powdered and without caking to seed.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201510942248.4A CN105474831B (en) | 2015-12-16 | 2015-12-16 | A kind of bletilla striata naked seed sterilization method |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201510942248.4A CN105474831B (en) | 2015-12-16 | 2015-12-16 | A kind of bletilla striata naked seed sterilization method |
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| CN105474831A CN105474831A (en) | 2016-04-13 |
| CN105474831B true CN105474831B (en) | 2017-11-17 |
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| CN105941392B (en) * | 2016-05-03 | 2018-10-09 | 湖南农业大学 | A kind of bletilla striata capsule store method suitable for tissue-culturing rapid propagation |
| CN106171974A (en) * | 2016-07-04 | 2016-12-07 | 广西大学 | The sterilization of Pseudobulbus Bletillae (Rhizoma Bletillae) the surface of the seed and the method for inoculation |
| CN112514751B (en) * | 2020-11-30 | 2022-06-17 | 广西壮族自治区亚热带作物研究所（广西亚热带农产品加工研究所） | Long-term storage and recovery method of cassava crossbreeding capsule |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP5148115B2 (en) * | 2004-02-06 | 2013-02-20 | ブリストル−マイヤーズ スクイブ カンパニー | Method for producing kinase inhibitor 2-aminothiazole-5-aromatic carboxamide compound |
| CN103081605A (en) * | 2012-12-30 | 2013-05-08 | 浙江中医药大学 | Method for collecting and storing bletilla striata seeds |
| CN104718841A (en) * | 2015-02-26 | 2015-06-24 | 李泽诚 | Treatment method of rhizoma bletillae seeds |
| CN104885936A (en) * | 2015-05-12 | 2015-09-09 | 陕西理工学院 | Seed liquid culture-based method used for rapid propagation of rhizoma bletillae seedlings |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH05148115A (en) * | 1991-07-08 | 1993-06-15 | Sanko Kk | Simple process for sowing, sprouting and transplantation of orchid |
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2015
- 2015-12-16 CN CN201510942248.4A patent/CN105474831B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP5148115B2 (en) * | 2004-02-06 | 2013-02-20 | ブリストル−マイヤーズ スクイブ カンパニー | Method for producing kinase inhibitor 2-aminothiazole-5-aromatic carboxamide compound |
| CN103081605A (en) * | 2012-12-30 | 2013-05-08 | 浙江中医药大学 | Method for collecting and storing bletilla striata seeds |
| CN104718841A (en) * | 2015-02-26 | 2015-06-24 | 李泽诚 | Treatment method of rhizoma bletillae seeds |
| CN104885936A (en) * | 2015-05-12 | 2015-09-09 | 陕西理工学院 | Seed liquid culture-based method used for rapid propagation of rhizoma bletillae seedlings |
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