CN107711503A - A kind of blood aspidistra method for tissue culture - Google Patents
A kind of blood aspidistra method for tissue culture Download PDFInfo
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- CN107711503A CN107711503A CN201711095284.7A CN201711095284A CN107711503A CN 107711503 A CN107711503 A CN 107711503A CN 201711095284 A CN201711095284 A CN 201711095284A CN 107711503 A CN107711503 A CN 107711503A
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- culture
- blood aspidistra
- sulfate
- seed
- aspidistra
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- 241001532036 Aspidistra Species 0.000 title claims abstract description 35
- 239000008280 blood Substances 0.000 title claims abstract description 35
- 210000004369 blood Anatomy 0.000 title claims abstract description 35
- 238000000034 method Methods 0.000 title claims abstract description 18
- 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
- 239000001963 growth medium Substances 0.000 claims abstract description 11
- 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
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims abstract description 8
- 235000019796 monopotassium phosphate Nutrition 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
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 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
- 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
- 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
- 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
- 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
- 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
- 230000001954 sterilising effect Effects 0.000 claims description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims 6
- 229910021529 ammonia Inorganic materials 0.000 claims 3
- 235000009508 confectionery Nutrition 0.000 claims 3
- 241000894006 Bacteria Species 0.000 claims 1
- 241000409198 Packera aurea Species 0.000 claims 1
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 abstract description 10
- 239000004471 Glycine Substances 0.000 abstract description 5
- 230000007226 seed germination Effects 0.000 abstract description 3
- 238000009395 breeding Methods 0.000 abstract description 2
- 230000001488 breeding effect Effects 0.000 abstract description 2
- PQMFVUNERGGBPG-UHFFFAOYSA-N (6-bromopyridin-2-yl)hydrazine Chemical compound NNC1=CC=CC(Br)=N1 PQMFVUNERGGBPG-UHFFFAOYSA-N 0.000 abstract 1
- 239000003153 chemical reaction reagent Substances 0.000 description 8
- 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 description 4
- 241000196324 Embryophyta Species 0.000 description 2
- 230000035784 germination Effects 0.000 description 2
- 238000007670 refining Methods 0.000 description 2
- 239000012882 rooting medium Substances 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 240000000001 Ludisia discolor Species 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 210000001367 artery Anatomy 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 238000004161 plant tissue culture Methods 0.000 description 1
- 230000001850 reproductive effect Effects 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 238000002054 transplantation Methods 0.000 description 1
- 210000003462 vein Anatomy 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
- A01H4/008—Methods for regeneration to complete plants
-
- 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
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 blood aspidistra method for tissue culture, including blood aspidistra seed culture to sprout into ball stem, strong seedling culture, root induction, four steps of hardening, and the seed culture medium that the blood aspidistra seed culture uses in sprouting into ball stem includes following component：Potassium nitrate 1600mg/L,Ammonium nitrate 1250mg/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,KI 0.83mg/L,Sodium molybdate 0.25mg/L,Na2‑EDTA37.3mg/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,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 blood aspidistra 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 blood aspidistra method for tissue culture.
Background technology
Blood aspidistra (Ludisia discolor (Ker-Gawl.) A.Rich.)：Plant is high 10-25 centimetres.Root-like stock is stretched
It is long, crawl, tool section.Stem is upright, has 3-4 pieces of leaf in nearly base portion.Blade is avette or ovate Long Circle, above blackish green, have 5 gold
Red glossiness arteries and veins, back side pale red；Have 2-3 pieces of pink sheath shape bract on stem on leaf.Raceme basidixed, tool
Several to more than 10 flowers；Floral white or with pale red, about 7 millimeters of diameter；The nearly semioval shape of petal, long 8-9 millimeters, wide 2-2.2 millis
Rice, tip are blunt.The month at florescence 2-4.At present, the breeding of blood aspidistra is mainly by division propagation or cutting propagation, in spring and autumn
Carry out, division propagation typically every plant division in 3 years once.But above-mentioned modes of reproduction reproductive efficiency is low, it is impossible to meet market need
Ask, therefore, it is necessary to blood aspidistra is quickly bred using plant tissue culture technique.But, existing blood aspidistra tissue training
Foster germination rate is relatively low, and yield is not high.
The content of the invention
It is an object of the invention to provide a kind of blood aspidistra method for tissue culture.
To realize the purpose of the present invention, the present invention uses following technical scheme：A kind of blood aspidistra method for tissue culture, including
Blood aspidistra seed culture is sprouted into ball stem, strong seedling culture, root induction, four steps of hardening, wherein the blood aspidistra seed
The seed culture medium that 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 |
| KI | 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 |
| KI | 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 training is seeded in after mature blood aspidistra seed is sterile-processed
Support and cultivated in base, turn green fortnight, continue culture 50-60 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 three minutes after with 75% alcohol disinfecting 30S, afterwards
With 35% dioxygen water sterilization 10min, 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 90-100 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 advantages below：
1. the present invention provides the method for tissue culture of blood aspidistra, blood aspidistra seed germination rate is high, more than 90%, acquisition
Seedling is sturdy, and axial root system is flourishing, and plantation survival rate is more than 90%.
2. sprouting of the seed culture medium to blood aspidistra seed has a major impact, seed culture medium provided by the invention can make
Blood aspidistra seed is sprouted 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
Blood aspidistra tissue cultures divide four-stage, are four seed sprouting, strong seedling culture, root induction and hardening ranks respectively
Section, so including the sprouting of blood aspidistra seed in blood aspidistra method for tissue culture provided by the invention turns into ball stem, strong sprout is raw
Root, four steps of hardening, specific incubation are as follows：
(1) seed, which is sprouted, turns into ball stem：Take mature blood aspidistra seed to carry out disinfection processing, rushed with running water plus liquid detergent
Wash 3 minutes, then 75% alcohol disinfecting 30 seconds, then with 35% dioxygen water sterilization 10 minutes, finally with aseptic water washing three times after
It is seeded in seed culture medium and cultivates, turn green a week, continue culture 20-30 days, is expanded by embryo, sprouts formation green circle
Bulb, 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 medium culture medium prescription
| Reagent name | Ormal weight (mg/L) | Reagent name | Ormal weight (mg/L) |
| Potassium nitrate | 1600 | Ferrous sulfate | 27.8 |
| Ammonium nitrate | 1250 | 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 |
| 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 | 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 |
| KI | 0.83 | Sucrose | 30000 |
| Sodium molybdate | 0.25 | ||
| Na2-EDTA | 37.3 |
3rd, 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, blood aspidistra seed germination rate can be made to reach more than 90%.Such as, NAA ormal weight is 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 is 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 blood aspidistra can be made to go out sturdy seedling.Such as, NAA ormal weight is 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 is 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 blood aspidistra method for tissue culture, including blood aspidistra seed culture are sprouted into ball stem, strong seedling culture, induction life
Root, four steps of hardening, it is characterized in that, the seed culture medium that the blood aspidistra seed culture uses in sprouting into ball stem includes
Following component：
Potassium nitrate 1600mg/L, ammonium nitrate 1250mg/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, KI 0.83mg/L, sodium molybdate 0.25mg/L, Na2-EDTA37.3mg/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. blood aspidistra method for tissue culture according to claim 2, it is characterized in that, the strong sprout used in the strong seedling culture
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, KI 0.83mg/L, sodium molybdate 0.25mg/L, Na2-EDTA37.3mg/L, ferrous sulfate 27.8mg/L, sweet ammonia
Sour 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. blood aspidistra method for tissue culture according to claim 1 or 2, it is characterized in that, used in the root induction
Root media includes following component：
Potassium nitrate 1650mg/L, ammonium nitrate 1350mg/L, magnesium sulfate 270mg/L, potassium dihydrogen phosphate 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-EDTA37.3mg/L, ferrous sulfate 27.8mg/L, sweet ammonia
Sour 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 powder
0.1-1mg/L。
4. blood aspidistra method for tissue culture according to claim 3, it is characterized in that, the seed culture is sprouted into ball stem
In step, it is seeded in seed culture medium and cultivates after mature blood aspidistra seed is sterile-processed, fortnight turns green, continues to cultivate
50-60 days, expanded by embryo, sprout and form green ball stem；Condition of culture is 25-26 DEG C, illumination 1600-1800lx,
Light application time is 10h/d.
5. blood aspidistra method for tissue culture according to claim 4, it is characterized in that, the described process of disinfecting is：From
Water add liquid detergent rinse 3 minutes after with 75% alcohol disinfecting 30 seconds, afterwards with 35% dioxygen water sterilization 10 minutes, then with nothing
Bacterium water flushes three times.
6. blood aspidistra method for tissue culture according to claim 3, it is characterized in that, in the strong sprout step, by the circle
Bulb, which is transferred in strong seedling culture base, carries out strong seedling culture, by 90-100 days strong seedling cultures, obtains the children that height reaches 1.5-3cm
Seedling, condition of culture：25-26 DEG C of temperature, illuminance 1900-2100lx, light application time 12h/d.
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| CN110250005A (en) * | 2019-07-22 | 2019-09-20 | 中国热带农业科学院热带作物品种资源研究所 | A kind of blood aspidistra cultivating method of new species |
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