CN107912300A - A kind of purple dendrobium method for tissue culture - Google Patents
A kind of purple dendrobium method for tissue culture Download PDFInfo
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- CN107912300A CN107912300A CN201711112797.4A CN201711112797A CN107912300A CN 107912300 A CN107912300 A CN 107912300A CN 201711112797 A CN201711112797 A CN 201711112797A CN 107912300 A CN107912300 A CN 107912300A
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- purple dendrobium
- culture
- sulfate
- potassium
- tissue culture
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- 241001523681 Dendrobium Species 0.000 title claims abstract description 35
- 238000000034 method Methods 0.000 title claims abstract description 21
- NLKNQRATVPKPDG-UHFFFAOYSA-M potassium iodide Chemical compound [K+].[I-] NLKNQRATVPKPDG-UHFFFAOYSA-M 0.000 claims abstract description 24
- 238000011218 seed culture Methods 0.000 claims abstract description 18
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims abstract description 16
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 claims abstract description 16
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 claims abstract description 16
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims abstract description 14
- 239000001963 growth medium Substances 0.000 claims abstract description 13
- PAWQVTBBRAZDMG-UHFFFAOYSA-N 2-(3-bromo-2-fluorophenyl)acetic acid Chemical compound OC(=O)CC1=CC=CC(Br)=C1F PAWQVTBBRAZDMG-UHFFFAOYSA-N 0.000 claims abstract description 8
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims abstract description 8
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 8
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims abstract description 8
- 229930006000 Sucrose Natural products 0.000 claims abstract description 8
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims abstract description 8
- 239000004327 boric acid Substances 0.000 claims abstract description 8
- 239000001110 calcium chloride Substances 0.000 claims abstract description 8
- 229910001628 calcium chloride Inorganic materials 0.000 claims abstract description 8
- 229910052799 carbon Inorganic materials 0.000 claims abstract description 8
- GVPFVAHMJGGAJG-UHFFFAOYSA-L cobalt dichloride Chemical compound [Cl-].[Cl-].[Co+2] GVPFVAHMJGGAJG-UHFFFAOYSA-L 0.000 claims abstract description 8
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 claims abstract description 8
- 239000000428 dust Substances 0.000 claims abstract description 8
- 239000011790 ferrous sulphate Substances 0.000 claims abstract description 8
- 235000003891 ferrous sulphate Nutrition 0.000 claims abstract description 8
- 230000006698 induction Effects 0.000 claims abstract description 8
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 claims abstract description 8
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims abstract description 8
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims abstract description 8
- 235000019341 magnesium sulphate Nutrition 0.000 claims abstract description 8
- 229940099596 manganese sulfate Drugs 0.000 claims abstract description 8
- 239000011702 manganese sulphate Substances 0.000 claims abstract description 8
- 235000007079 manganese sulphate Nutrition 0.000 claims abstract description 8
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims abstract description 8
- 235000001968 nicotinic acid Nutrition 0.000 claims abstract description 8
- 229960003512 nicotinic acid Drugs 0.000 claims abstract description 8
- 239000011664 nicotinic acid Substances 0.000 claims abstract description 8
- 239000004323 potassium nitrate Substances 0.000 claims abstract description 8
- 235000010333 potassium nitrate Nutrition 0.000 claims abstract description 8
- 239000011684 sodium molybdate Substances 0.000 claims abstract description 8
- 235000015393 sodium molybdate Nutrition 0.000 claims abstract description 8
- TVXXNOYZHKPKGW-UHFFFAOYSA-N sodium molybdate (anhydrous) Chemical compound [Na+].[Na+].[O-][Mo]([O-])(=O)=O TVXXNOYZHKPKGW-UHFFFAOYSA-N 0.000 claims abstract description 8
- 239000005720 sucrose Substances 0.000 claims abstract description 8
- 229960000344 thiamine hydrochloride Drugs 0.000 claims abstract description 8
- 235000019190 thiamine hydrochloride Nutrition 0.000 claims abstract description 8
- 239000011747 thiamine hydrochloride Substances 0.000 claims abstract description 8
- DPJRMOMPQZCRJU-UHFFFAOYSA-M thiamine hydrochloride Chemical compound Cl.[Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N DPJRMOMPQZCRJU-UHFFFAOYSA-M 0.000 claims abstract description 8
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 claims abstract description 8
- 229960001763 zinc sulfate Drugs 0.000 claims abstract description 8
- 229910000368 zinc sulfate Inorganic materials 0.000 claims abstract description 8
- 239000004471 Glycine Substances 0.000 claims abstract description 7
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims abstract description 7
- 235000019796 monopotassium phosphate Nutrition 0.000 claims abstract description 7
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims abstract description 7
- ZGTMUACCHSMWAC-UHFFFAOYSA-L EDTA disodium salt (anhydrous) Chemical compound [Na+].[Na+].OC(=O)CN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O ZGTMUACCHSMWAC-UHFFFAOYSA-L 0.000 claims abstract description 6
- 244000061456 Solanum tuberosum Species 0.000 claims abstract description 3
- 235000002595 Solanum tuberosum Nutrition 0.000 claims abstract description 3
- 239000000413 hydrolysate Substances 0.000 claims abstract description 3
- 229940064880 inositol 100 mg Drugs 0.000 claims abstract 2
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 claims description 7
- 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 claims description 6
- 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 claims description 6
- 229960000367 inositol Drugs 0.000 claims description 6
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 241000234295 Musa Species 0.000 claims description 5
- 235000018290 Musa x paradisiaca Nutrition 0.000 claims description 5
- 230000000249 desinfective effect Effects 0.000 claims description 5
- 238000004659 sterilization and disinfection Methods 0.000 claims description 5
- 230000001939 inductive effect Effects 0.000 claims description 4
- 239000000843 powder Substances 0.000 claims description 4
- 230000001954 sterilising effect Effects 0.000 claims description 4
- MYMOFIZGZYHOMD-UHFFFAOYSA-N Dioxygen Chemical compound O=O MYMOFIZGZYHOMD-UHFFFAOYSA-N 0.000 claims description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 3
- 239000003599 detergent Substances 0.000 claims description 3
- 229910001882 dioxygen Inorganic materials 0.000 claims description 3
- 238000005286 illumination Methods 0.000 claims description 3
- 239000007788 liquid Substances 0.000 claims description 3
- 210000001161 mammalian embryo Anatomy 0.000 claims description 3
- 230000008569 process Effects 0.000 claims description 3
- 239000008399 tap water Substances 0.000 claims description 3
- 235000020679 tap water Nutrition 0.000 claims description 3
- 238000005406 washing Methods 0.000 claims description 3
- 230000015572 biosynthetic process Effects 0.000 claims description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims 2
- 239000011734 sodium Substances 0.000 claims 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-L Phosphate ion(2-) Chemical compound OP([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-L 0.000 claims 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 claims 1
- 229910021529 ammonia Inorganic materials 0.000 claims 1
- 235000009508 confectionery Nutrition 0.000 claims 1
- 239000011591 potassium Substances 0.000 claims 1
- 229910052700 potassium Inorganic materials 0.000 claims 1
- 230000007226 seed germination Effects 0.000 abstract description 3
- PQMFVUNERGGBPG-UHFFFAOYSA-N (6-bromopyridin-2-yl)hydrazine Chemical compound NNC1=CC=CC(Br)=N1 PQMFVUNERGGBPG-UHFFFAOYSA-N 0.000 abstract 1
- 238000009395 breeding Methods 0.000 abstract 1
- 230000001488 breeding effect Effects 0.000 abstract 1
- 239000003153 chemical reaction reagent Substances 0.000 description 8
- 241000196324 Embryophyta Species 0.000 description 4
- 241001264165 Dendrobium devonianum Species 0.000 description 3
- 240000004638 Dendrobium nobile Species 0.000 description 3
- 230000004083 survival effect Effects 0.000 description 3
- 230000035784 germination Effects 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 238000007670 refining Methods 0.000 description 2
- 239000012882 rooting medium Substances 0.000 description 2
- 241000208140 Acer Species 0.000 description 1
- 235000013361 beverage Nutrition 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000002513 implantation Methods 0.000 description 1
- 230000002073 mitogenetic effect Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000002269 spontaneous effect Effects 0.000 description 1
- 239000004575 stone Substances 0.000 description 1
- 238000002054 transplantation Methods 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
- A01H4/001—Culture apparatus for tissue culture
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a kind of purple dendrobium method for tissue culture, sprouted including purple dendrobium seed culture into ball stem, strong seedling culture, root induction, four steps of hardening, the seed culture medium that the purple dendrobium seed culture uses in sprouting into ball stem include following component：Potassium nitrate 1200mg/L,Ammonium nitrate 1050mg/L,Magnesium sulfate 370mg/L,Potassium dihydrogen phosphate 170mg/L,Calcium chloride 440mg/L,Copper sulphate 0.025mg/L,Cobalt chloride 0.025mg/L,Manganese sulfate 16.9mg/L,Zinc sulfate 8.6mg/L,Boric acid 6.2mg/L,Potassium iodide 0.83mg/L,Sodium molybdate 0.25mg/L,Na2‑EDTA 37.3mg/L,Ferrous sulfate 21.8mg/L,Glycine 2.0mg/L,Thiamine hydrochloride 0.1mg/L,Puridoxine hydrochloride 0.5mg/L,Nicotinic acid 0.5mg/L,Inositol 100mg/L,NAA 1‑10mg/L,6‑BA 5‑10mg/L,Potato 100000mg/L,Carbon dust 250mg/L,Sucrose 30000mg/L,Lactoalbumin hydrolysate 700mg/L.The purple dendrobium seed germination rate of this method is high, and more than 90%, breeding potential is greatly improved.
Description
Technical field
The present invention relates to a kind of purple dendrobium method for tissue culture.
Background technology
Purple dendrobium is a kind of orchid family Dendrobium Sw using dendrobium devonianum as representative, is the one of the rare medicinal plant stem of noble dendrobium
A kind.Dendrobium devonianum also known as purple skin are blue, are orchid dendrobium devonianum Dendrobium devoninum Paxt..Happiness temperature
It is warm, moistening, the environment of half cloudy half light.Plant of the purple dendrobium as medicine-food two-purpose, can eat raw, can also add through manually twisting
Work is into purple skin maple bucket.In addition, beverage, tablet, capsule, electuary etc. can be made by deep processing in purple dendrobium.
The propagation method of purple dendrobium seedling is divided to generative propagation and vegetative propagation two major classes.In the spontaneous growth process of the stem of noble dendrobium
In, based on generative propagation, supplemented by vegetative propagation.Artificial cultivation is then based on vegetative propagation.Vegetative propagation refers to utilize seedling,
Cane, bud and a certain organ of plant, grow up to new plant.Tissue culture propagation method is one kind of vegetative propagation.Current tissue culture is numerous
The method of growing is to strip the growing point of stem of noble dendrobium stem apex and bud, and minority is using a stem bar bud part, after strict sterilization, in sterile condition
In lower implantation culture medium, through the culture of 2~3 months, in incubation, seedling was constantly mitogenetic, became 2, the 2 gradual expansions become with 1
Greatly, when seedling covers with culture medium, further take out point and to plant, be known as subculture or expand it is numerous, by expand several times it is numerous after, then change culture medium into
Point, promote it and take root, when offspring to be generated grows to certain height, you can take out and carry out hardening.But current tissue culture propagation exists
Germination rate is low and the problem of rooting rate is low, and the survival rate of seedling is very low.
The content of the invention
It is an object of the invention to provide a kind of purple dendrobium method for tissue culture.
To achieve the object of the present invention, the present invention uses following technical scheme：A kind of purple dendrobium method for tissue culture, bag
Purple dendrobium seed culture is included to sprout into ball stem, strong seedling culture, root induction, four steps of hardening, wherein, the purple skin stone
The seed culture medium that dry measure used in former times seed culture uses in sprouting into ball stem includes following component：
First, seed culture based formulas
The strong seedling culture base used in the strong seedling culture includes following component：
2nd, strong seedling culture based formulas
| Reagent name | Ormal weight (mg/L) | Reagent name | Ormal weight (mg/L) |
| Potassium nitrate | 2000 | Ferrous sulfate | 27.8 |
| Ammonium nitrate | 1750 | Glycine | 2.0 |
| Magnesium sulfate | 370 | Thiamine hydrochloride | 0.2 |
| Potassium dihydrogen phosphate | 170 | Puridoxine hydrochloride | 0.65 |
| Calcium chloride | 550 | Nicotinic acid | 0.5 |
| Copper sulphate | 0.025 | Inositol | 50 |
| Cobalt chloride | 0.025 | NAA | 1-10 |
| Manganese sulfate | 16.9 | 6-BA | 5-10 |
| Zinc sulfate | 8.6 | Banana | 200000 |
| Boric acid | 6.2 | Carbon dust | 500 |
| Potassium iodide | 0.83 | Sucrose | 30000 |
| Sodium molybdate | 0.25 | ||
| Na2-EDTA | 37.3 |
The root media used in the root induction includes following component：
3rd, prescription of rooting medium
| Reagent name | Ormal weight (mg/L) | Reagent name | Ormal weight (mg/L) |
| Potassium nitrate | 1650 | Ferrous sulfate | 27.8 |
| Ammonium nitrate | 1350 | Glycine | 2.0 |
| Magnesium sulfate | 270 | Thiamine hydrochloride | 0.2 |
| Potassium dihydrogen phosphate | 130 | Puridoxine hydrochloride | 0.65 |
| Calcium chloride | 450 | Nicotinic acid | 0.5 |
| Copper sulphate | 0.025 | Inositol | 5 |
| Cobalt chloride | 0.025 | NAA | 1-10 |
| Manganese sulfate | 16.9 | 6-BA | 0.1-10 |
| Zinc sulfate | 8.6 | Banana | 100000 |
| Boric acid | 6.2 | Carbon dust | 500 |
| Potassium iodide | 0.83 | Sucrose | 18000 |
| Sodium molybdate | 0.25 | Ai Bidi root-inducing powders | 0.1-1 |
| Na2-EDTA | 37.3 |
The seed culture is sprouted into ball stem step, and seed is seeded in after ripe purple dendrobium seed is sterile-processed
Cultivated in culture medium, turn green a week, continue culture 20-30 days, expanded by embryo, sprout the green ball stem of formation；Culture
Condition is 25-26 DEG C, illumination 1600-18001x, light application time 10h/d.
The process of disinfecting is：Tap water add liquid detergent rinse 3 minutes after with 75% alcohol disinfecting 30 seconds, afterwards
With 35% dioxygen water sterilization 10 minutes, then with aseptic water washing three times.
In the strong sprout step, ball stem, which is transferred in strong seedling culture base, carries out strong seedling culture, is trained by 80-90 days strong sprouts
Support, obtain the seedling that height reaches 1.5-3cm, condition of culture：25-26 DEG C of temperature, illuminance 1900-2100lx, light application time
For 12h/d.
The present invention has the following advantages：
1. the present invention provides the method for tissue culture of purple dendrobium, purple dendrobium seed germination rate is high, more than 90%, obtains
The seedling obtained is sturdy, and axial root system is flourishing, and plantation survival rate is more than 90%.
2. sprouting of the seed culture medium to purple dendrobium seed has a major impact, seed culture medium provided by the invention can be with
Purple dendrobium seed is set to sprout more than 90%.
3. the strong seedling culture base that the present invention uses can promote ball stem to generate sturdy seedling.
4. the root media that the present invention uses can promote seedling to take root, the axial root system of gained is flourishing, is conducive to refining
Survived in seedling and plantation.
Embodiment
With reference to specific embodiment, the present invention is described in further detail, but does not form any limit to the present invention
System, any limited modification made within the scope of the invention as claimed, still in the claims of the present invention.
Embodiment 1
Purple dendrobium tissue cultures divide four-stage, are seed sprouting, strong seedling culture, root induction and hardening four respectively
In the stage, so including the sprouting of purple dendrobium seed in purple dendrobium method for tissue culture provided by the invention becomes ball stem, strengthen
Seedling culture, takes root, four steps of hardening, and specific incubation is as follows：
(1) seed, which is sprouted, becomes ball stem：Ripe purple dendrobium seed is taken to carry out disinfection processing, with tap water plus liquid detergent
Rinse 3 minutes, then 75% alcohol disinfecting 30 seconds, then with 35% dioxygen water sterilization 10 minutes, finally aseptic water washing with three times
After be seeded in seed culture medium and cultivate, turn green a week, continue culture 20-30 days, expanded by embryo, sprouts formed it is green
Ball stem, the germination rate of seed is more than 90%.Condition of culture is 25-26 DEG C, illumination 1600-1800lx, and light application time is
10h/d。
(2) strong seedling culture：The ball stem produced in step (1) is transferred in strong seedling culture base and carries out strong seedling culture, is passed through
80-90 days strong seedling cultures, the sturdy seedling for highly reaching 1.5-3cm occur, condition of culture：25-26 DEG C of temperature, illuminance
1900-2100lx, light application time 12h/d.
(3) root induction：The seedling that step (2) obtains is transferred to root media and carries out root induction, is grown up to hair
Up to the rooted seedling of axial root system.
(4) hardening：The rooted seedling that step (3) is obtained carries out hardening domestication, adapts it to external growth environment.Complete refining
After seedling, gained domestication transplantation of seedlings greenhouse or outdoor, while Cultivate administration, domestication shoot survival percent is more than 90%.
Seed culture based formulas
| Reagent name | Ormal weight (mg/L) | Reagent name | Ormal weight (mg/L) |
| Potassium nitrate | 1200 | Ferrous sulfate | 21.8 |
| Ammonium nitrate | 1050 | Glycine | 2.0 |
| Magnesium sulfate | 370 | Thiamine hydrochloride | 0.1 |
| Potassium dihydrogen phosphate | 170 | Puridoxine hydrochloride | 0.5 |
| Calcium chloride | 440 | Nicotinic acid | 0.5 |
| Copper sulphate | 0.025 | Inositol | 100 |
| Cobalt chloride | 0.025 | NAA | 1-10 |
| Manganese sulfate | 16.9 | 6-BA | 5-10 |
| Zinc sulfate | 8.6 | Potato | 100000 |
| Boric acid | 6.2 | Carbon dust | 250 |
| Potassium iodide | 0.83 | Sucrose | 30000 |
| Sodium molybdate | 0.25 | Lactoalbumin hydrolysate | 700 |
| Na2-EDTA | 37.3 |
Strong seedling culture based formulas
| Reagent name | Ormal weight (mg/L) | Reagent name | Ormal weight (mg/L) |
| Potassium nitrate | 2000 | Ferrous sulfate | 27.8 |
| Ammonium nitrate | 1750 | Glycine | 2.0 |
| Magnesium sulfate | 370 | Thiamine hydrochloride | 0.2 |
| Potassium dihydrogen phosphate | 170 | Puridoxine hydrochloride | 0.65 |
| Calcium chloride | 550 | Nicotinic acid | 0.5 |
| Copper sulphate | 0.025 | Inositol | 50 |
| Cobalt chloride | 0.025 | NAA | 1-10 |
| Manganese sulfate | 16.9 | 6-BA | 5-10 |
| Zinc sulfate | 8.6 | Banana | 200000 |
| Boric acid | 6.2 | Carbon dust | 500 |
| Potassium iodide | 0.83 | Sucrose | 30000 |
| Sodium molybdate | 0.25 | ||
| Na2-EDTA | 37.3 |
Prescription of rooting medium
The ormal weight of NAA in seed culture medium adjusts in the range of 1-10mg/L, and the ormal weight of 6-BA is in 5-10mg/L
In the range of adjust, purple dendrobium seed germination rate can be made to reach more than 90%.Such as, the ormal weight of NAA for 1mg/L,
1.5mg/L、2mg/L、2.5mg/L、3mg/L、3.5mg/L、4mg/L、4.5mg/L、5mg/L、5.5mg/L、6mg/L、6.5mg/
L, 7mg/L, 7.5mg/L, 8mg/L, 8.5mg/L, 9mg/L, 9.5mg/L and 10mg/L.The ormal weight of 6-BA for 5mg/L,
5.5mg/L, 6mg/L, 6.5mg/L, 7mg/L, 7.5mg/L, 8mg/L, 8.5mg/L, 9mg/L, 9.5mg/L and 10mg/L.
The ormal weight of NAA in strong seedling culture base adjusts in the range of 1-10mg/L, and the ormal weight of 6-BA is in 5-10mg/L
In the range of adjust, the ball stem length of purple dendrobium can be made to go out sturdy seedling.Such as, the ormal weight of NAA for 1mg/L,
1.5mg/L、2mg/L、2.5mg/L、3mg/L、3.5mg/L、4mg/L、4.5mg/L、5mg/L、5.5mg/L、6mg/L、6.5mg/
L, 7mg/L, 7.5mg/L, 8mg/L, 8.5mg/L, 9mg/L, 9.5mg/L and 10mg/L.The ormal weight of 6-BA for 5mg/L,
5.5mg/L, 6mg/L, 6.5mg/L, 7mg/L, 7.5mg/L, 8mg/L, 8.5mg/L, 9mg/L, 9.5mg/L and 10mg/L.
The ormal weight of NAA in root media adjusts in the range of 1-10mg/L, and the ormal weight of 6-BA is in 5-10mg/L
In the range of adjust, the ormal weight of Ai Bidi root-inducing powders adjusts in the range of 0.1-1mg/L, seedling grow prosperity axial root system.Such as,
The ormal weight of NAA for 1mg/L, 1.5mg/L, 2mg/L, 2.5mg/L, 3mg/L, 3.5mg/L, 4mg/L, 4.5mg/L, 5mg/L,
5.5mg/L, 6mg/L, 6.5mg/L, 7mg/L, 7.5mg/L, 8mg/L, 8.5mg/L, 9mg/L, 9.5mg/L and 10mg/L.
The ormal weight of 6-BA for 5mg/L, 5.5mg/L, 6mg/L, 6.5mg/L, 7mg/L, 7.5mg/L, 8mg/L, 8.5mg/L, 9mg/L,
9.5mg/L and 10mg/L.The ormal weight of Ai Bidi root-inducing powders for 0.1mg/L, 0.2mg/L, 0.3mg/L, 0.4mg/L,
0.5mg/L, 0.6mg/L, 0.7mg/L, 0.8mg/L, 0.9mg/L, 1mg/L.
Claims (6)
1. a kind of purple dendrobium method for tissue culture, including purple dendrobium seed culture are sprouted into ball stem, strong seedling culture, induction
Take root, four steps of hardening, it is characterized in that, the seed culture medium that the purple dendrobium seed culture uses in sprouting into ball stem
Include following component：
Potassium nitrate 1200mg/L, ammonium nitrate 1050mg/L, magnesium sulfate 370mg/L, potassium dihydrogen phosphate 170mg/L, calcium chloride
440mg/L, copper sulphate 0.025mg/L, cobalt chloride 0.025mg/L, manganese sulfate 16.9mg/L, zinc sulfate 8.6mg/L, boric acid
6.2mg/L, potassium iodide 0.83mg/L, sodium molybdate 0.25mg/L, Na2-EDTA 37.3mg/L, ferrous sulfate 21.8mg/L, sweet ammonia
Sour 2.0mg/L, thiamine hydrochloride 0.1mg/L, puridoxine hydrochloride 0.5mg/L, nicotinic acid 0.5mg/L, inositol 100mg/L, NAA 1-
10mg/L, 6-BA 5-10mg/L, potato 100000mg/L, carbon dust 250mg/L, sucrose 30000mg/L, lactoalbumin hydrolysate
700mg/L。
2. purple dendrobium method for tissue culture according to claim 1, it is characterized in that, what is used in the strong seedling culture is strong
Seedling culture medium includes following component：Potassium nitrate 2000mg/L, ammonium nitrate 1750mg/L, magnesium sulfate 370mg/L, potassium dihydrogen phosphate
170mg/L, calcium chloride 550mg/L, copper sulphate 0.025mg/L, cobalt chloride 0.025mg/L, manganese sulfate 16.9mg/L, zinc sulfate
8.6mg/L, boric acid 6.2mg/L, potassium iodide 0.83mg/L, sodium molybdate 0.25mg/L, Na2- EDTA 37.3mg/L, ferrous sulfate
27.8mg/L, glycine 2.0mg/L, thiamine hydrochloride 0.2mg/L, puridoxine hydrochloride 0.65mg/L, nicotinic acid 0.5mg/L, inositol
50mg/L, NAA 1-10mg/L, 6-BA 5-10mg/L, banana 200000mg/L, carbon dust 500mg/L, sucrose 30000mg/L.
3. purple dendrobium method for tissue culture according to claim 1 or 2, it is characterized in that, used in the root induction
Root media include following component：Potassium nitrate 1650mg/L, ammonium nitrate 1350mg/L, magnesium sulfate 270mg/L, biphosphate
Potassium 130mg/L, calcium chloride 450mg/L, copper sulphate 0.025mg/L, cobalt chloride 0.025mg/L, manganese sulfate 16.9mg/L, zinc sulfate
8.6mg/L, boric acid 6.2mg/L, potassium iodide 0.83mg/L, sodium molybdate 0.25mg/L, Na2- EDTA 37.3mg/L, ferrous sulfate
27.8mg/L, glycine 2.0mg/L, thiamine hydrochloride 0.2mg/L, puridoxine hydrochloride 0.65mg/L, nicotinic acid 0.5mg/L, inositol
5mg/L, NAA 1-10mg/L, 6-BA 0.1-10mg/L, banana 100000mg/L, carbon dust 500mg/L, sucrose 18000mg/L,
Ai Bidi root-inducing powders 0.1-1mg/L.
4. purple dendrobium method for tissue culture according to claim 3, it is characterized in that, the seed culture is sprouted into ball
In stem step, it is seeded in seed culture medium and cultivates after ripe purple dendrobium seed is sterile-processed, turns green a week, continue to train
Support 20-30 days, expanded by embryo, sprout the green ball stem of formation；Condition of culture is 25-26 DEG C, illumination 1600-
1800lx, light application time 10h/d.
5. purple dendrobium method for tissue culture according to claim 4, it is characterized in that, the process of disinfecting is：
Tap water add liquid detergent rinse 3 minutes after with 75% alcohol disinfecting 30 seconds, afterwards with 35% dioxygen water sterilization 10 minutes, Ran Houyong
Aseptic water washing is three times.
6. purple dendrobium method for tissue culture according to claim 4, it is characterized in that, in the strong sprout step, ball stem
It is transferred in strong seedling culture base and carries out strong seedling culture, by 80-90 days strong seedling cultures, obtains the seedling that height reaches 1.5-3cm, training
The condition of supporting：25-26 DEG C of temperature, illuminance 1900-2100lx, light application time 12h/d.
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