CN107736247A - A kind of protruding canine teeth stem of noble dendrobium method for tissue culture - Google Patents
A kind of protruding canine teeth stem of noble dendrobium method for tissue culture Download PDFInfo
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- CN107736247A CN107736247A CN201711112723.0A CN201711112723A CN107736247A CN 107736247 A CN107736247 A CN 107736247A CN 201711112723 A CN201711112723 A CN 201711112723A CN 107736247 A CN107736247 A CN 107736247A
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- stem
- noble dendrobium
- canine teeth
- protruding canine
- culture
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- 240000004638 Dendrobium nobile Species 0.000 title claims abstract description 35
- 210000003464 cuspid Anatomy 0.000 title claims abstract description 35
- 238000000034 method Methods 0.000 title claims abstract description 17
- 238000011218 seed culture Methods 0.000 claims abstract description 17
- 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 12
- 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
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-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
- 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
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 9
- 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
- 235000018290 Musa x paradisiaca Nutrition 0.000 claims description 5
- 230000000249 desinfective effect Effects 0.000 claims description 5
- 238000005286 illumination Methods 0.000 claims description 4
- 230000001939 inductive effect Effects 0.000 claims description 4
- 239000000843 powder Substances 0.000 claims description 4
- 238000004659 sterilization and disinfection Methods 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
- 239000007788 liquid Substances 0.000 claims description 3
- 210000001161 mammalian embryo Anatomy 0.000 claims description 3
- 230000001954 sterilising effect Effects 0.000 claims description 3
- 238000005406 washing Methods 0.000 claims description 3
- 230000008569 process Effects 0.000 claims description 2
- 239000011734 sodium Substances 0.000 claims 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims 2
- 240000008790 Musa x paradisiaca Species 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
- 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 abstract description 5
- 230000001850 reproductive effect Effects 0.000 abstract description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 8
- 241000234295 Musa Species 0.000 description 3
- 230000004083 survival effect Effects 0.000 description 3
- 238000007670 refining Methods 0.000 description 2
- 239000012882 rooting medium Substances 0.000 description 2
- 230000007226 seed germination Effects 0.000 description 2
- 239000004575 stone Substances 0.000 description 2
- 241000372847 Dendrobium monticola Species 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000004927 clay Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000003337 fertilizer Substances 0.000 description 1
- 230000035784 germination Effects 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 238000004161 plant tissue culture Methods 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 239000011435 rock Substances 0.000 description 1
- 238000002054 transplantation Methods 0.000 description 1
- 230000017260 vegetative to reproductive phase transition of meristem Effects 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
Abstract
The invention discloses a kind of protruding canine teeth stem of noble dendrobium method for tissue culture, sprouted including protruding canine teeth stem of noble dendrobium seed culture into ball stem, strong seedling culture, root induction, four steps of hardening, the seed culture medium that the protruding canine teeth stem of noble dendrobium seed culture uses in sprouting into ball stem include following component：Potassium nitrate 1700mg/L, ammonium nitrate 1350mg/L, magnesium sulfate 360mg/L, potassium dihydrogen phosphate 160mg/L, calcium chloride 430mg/L, copper sulphate 0.025mg/L, cobalt chloride 0.025mg/L, manganese sulfate 15.9mg/L, zinc sulfate 7.6mg/L, boric acid 5.2mg/L, KI 0.83mg/L, sodium molybdate 0.25mg/L, Na2EDTA 37.3mg/L, ferrous sulfate 27.8mg/L, glycine 2.0mg/L, thiamine hydrochloride 0.1mg/L, puridoxine hydrochloride 0.5mg/L, nicotinic acid 0.5mg/L, the 10mg/L of inositol 100mg/L, NAA 1, the 10mg/L of 6 BA 5, potato 100000mg/L, carbon dust 250mg/L, sucrose 30000mg/L, lactoalbumin hydrolysate 700mg/L.The present invention improves the reproductive efficiency of protruding canine teeth stem of noble dendrobium tissue cultures.
Description
Technical field
The present invention relates to a kind of protruding canine teeth stem of noble dendrobium method for tissue culture.
Background technology
The protruding canine teeth stem of noble dendrobium, that is, hide the southern stem of noble dendrobium (scientific name：Dendrobium monticola P.F.Hunt et Summerh.) it is raw
In on the mountain valley rock of height above sea level 1750-2200 rice.Guangxi China southwest and southern Tibet are distributed in, the India northwestward is through Nepal
It is also distributed to Sillim and Thailand.It has abundant active polysaccharide, has medical value.It generally has raceme normal
1-4, basidixed or sends from the stem top of raw tool leaf then, it is near upright or it is curved hang down, it is long 2.5-5 centimetres, have several little Hua；The flowers are in blossom
Exhibition, white；The long and narrow circle of petal, tip is tapering, contracting of slightly being hung in the middle part of the nearly ellipse of lip, and more than middle part 3 split, base portion tool shortly clawed,
Great sight, there is higher horticultural importance.
The protruding canine teeth stem of noble dendrobium be it is a kind of like shady and cool herbaceos perennial, like it is warm, moist, with 1000 millimeters of annual rainfall
Above, the environment of half cloudy half light, 1 monthly mean temperature grows in the subtropical zone remote, thickly forested mountains higher than 8 DEG C to be preferred, suitable growth temperature
For 15 to 28 degree, suitable growth air humidity is more than 60%, requires not very strict to clay fertilizer, wild more in loose and thick tree
Grown on skin or trunk, some is also grown in crack of stone.Belong to aerial root system, major requirement root permeability is good, the matrix of use
Preferably ventilated drainage, under suitable temperature humidity, the speed of growth is fast, and survival ability is very strong.Annual the end of spring and the beginning of summer,
Inflorescence is extracted out on biennial stem top section, and Post flowering grows sprouting from stem foot and develops into stem, and autumn and winter enters rest period.Mesh
Before, the artificial culture of the protruding canine teeth stem of noble dendrobium includes division propagation, cutting propagation, proliferation, still, above-mentioned modes of reproduction reproductive efficiency
It is low, it is impossible to meet the market demand, therefore, it is necessary to the protruding canine teeth stem of noble dendrobium is quickly bred using plant tissue culture technique.
The content of the invention
It is an object of the invention to provide a kind of protruding canine teeth stem of noble dendrobium method for tissue culture.
To realize the purpose of the present invention, the present invention uses following technical scheme：A kind of protruding canine teeth stem of noble dendrobium method for tissue culture, bag
Protruding canine teeth stem of noble dendrobium seed culture is included to sprout into ball stem, strong seedling culture, root induction, four steps of hardening, wherein the protruding canine teeth stone
The culture medium difference that dry measure used in former times seed culture uses in sprouting into ball stem, strong sprout and root induction is as follows：
First, seed culture medium culture medium prescription
2nd, strong seedling culture based formulas
| Reagent name | Ormal weight (mg/L) | Reagent name | Ormal weight (mg/L) |
| Potassium nitrate | 1900 | Ferrous sulfate | 27.8 |
| Ammonium nitrate | 1650 | 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 |
| KI | 0.83 | Sucrose | 30000 |
| Sodium molybdate | 0.25 | ||
| Na2-EDTA | 37.3 |
3rd, prescription of rooting medium
| Reagent name | Ormal weight (mg/L) | Reagent name | Ormal weight (mg/L) |
| Potassium nitrate | 1550 | Ferrous sulfate | 27.8 |
| Ammonium nitrate | 1250 | Glycine | 2.0 |
| Magnesium sulfate | 260 | 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 |
| KI | 0.83 | Sucrose | 18000 |
| Sodium molybdate | 0.25 | Ai Bidi root-inducing powders | 0.1-1 |
| Na2-EDTA | 35.3 |
The seed culture is sprouted into ball stem step, and seed is seeded in after ripe protruding canine teeth stem of noble dendrobium seed is sterile-processed
Cultivated in culture medium, turn green a week, continue culture 20-30 days, expanded by embryo, sprout and form green ball stem；Culture
Condition is 25-26 DEG C, illumination 1600-1800lx, light application time 10h/d.
The described process of disinfecting is：Running 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 seedling culture step, ball stem, which is transferred in strong seedling culture base, carried out strong seedling culture, by 80-90 days strong sprouts
Culture, obtain the seedling that height reaches 1.5-3cm, condition of culture：25-26 DEG C of temperature, illuminance 1900-2100lx, during illumination
Between be 12h/d.
The present invention has advantages below：
1. the present invention provides the method for tissue culture of the protruding canine teeth stem of noble dendrobium, protruding canine teeth stem of noble 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 greatly improves reproductive efficiency more than 90%.
2. sprouting of the seed culture medium to protruding canine teeth stem of noble dendrobium seed has a major impact, seed culture medium provided by the invention can be with
Protruding canine teeth stem of noble 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 advantageous 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
Protruding canine teeth stem of noble 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 protruding canine teeth stem of noble dendrobium seed in protruding canine teeth stem of noble dendrobium method for tissue culture provided by the invention turns into ball stem, strengthen
Seedling, take root, four steps of hardening, specific incubation is as follows：
(1) seed, which is sprouted, turns into ball stem：Ripe protruding canine teeth stem of noble dendrobium seed is taken to carry out disinfection processing, with running 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, to sprout formation 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：Caused ball stem 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 | 1700 | Ferrous sulfate | 27.8 |
| Ammonium nitrate | 1350 | Glycine | 2.0 |
| Magnesium sulfate | 360 | Thiamine hydrochloride | 0.1 |
| Potassium dihydrogen phosphate | 160 | Puridoxine hydrochloride | 0.5 |
| Calcium chloride | 430 | Nicotinic acid | 0.5 |
| Copper sulphate | 0.025 | Inositol | 100 |
| Cobalt chloride | 0.025 | NAA | 1-10 |
| Manganese sulfate | 15.9 | 6-BA | 5-10 |
| Zinc sulfate | 7.6 | Potato | 100000 |
| Boric acid | 5.2 | Carbon dust | 250 |
| KI | 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 | 1900 | Ferrous sulfate | 27.8 |
| Ammonium nitrate | 1650 | 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 |
| KI | 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 6-BA ormal weight is in 5-10mg/L
In the range of adjust, protruding canine teeth stem of noble dendrobium seed germination rate can be made to reach more than 90%.Such as, NAA ormal weight be 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.6-BA ormal weight be 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 6-BA ormal weight is in 5-10mg/L
In the range of adjust, the ball stem length of the protruding canine teeth stem of noble dendrobium can be made to go out sturdy seedling.Such as, NAA ormal weight be 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.6-BA ormal weight be 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 6-BA ormal weight 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,
NAA ormal weight be 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.
6-BA ormal weight be 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 be 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 protruding canine teeth stem of noble dendrobium method for tissue culture, including protruding canine teeth stem of noble 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 protruding canine teeth stem of noble dendrobium seed culture uses in sprouting into ball stem
Include following component：
Potassium nitrate 1700mg/L, ammonium nitrate 1350mg/L, magnesium sulfate 360mg/L, potassium dihydrogen phosphate 160mg/L, calcium chloride
430mg/L, copper sulphate 0.025mg/L, cobalt chloride 0.025mg/L, manganese sulfate 15.9mg/L, zinc sulfate 7.6mg/L, boric acid
5.2mg/L, KI 0.83mg/L, sodium molybdate 0.25mg/L, Na2- EDTA 37.3mg/L, ferrous sulfate 27.8mg/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. protruding canine teeth stem of noble 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 1900mg/L, ammonium nitrate 1650mg/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, KI 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. protruding canine teeth stem of noble 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 1550mg/L, ammonium nitrate 1250mg/L, magnesium sulfate 260mg/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, KI 0.83mg/L, sodium molybdate 0.25mg/L, Na2- EDTA 35.3mg/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. protruding canine teeth stem of noble 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 protruding canine teeth stem of noble dendrobium seed is sterile-processed, turns green a week, continue to train
Support 20-30 days, expanded by embryo, sprout and form green ball stem；Condition of culture is 25-26 DEG C, illumination 1600-
1800lx, light application time 10h/d.
5. protruding canine teeth stem of noble dendrobium method for tissue culture according to claim 4, it is characterized in that, the described process of disinfecting is：
Running 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. protruding canine teeth stem of noble 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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