CN115191352A - Preparation method of Louisiana iris hydroponic aseptic seedlings - Google Patents
Preparation method of Louisiana iris hydroponic aseptic seedlings Download PDFInfo
- Publication number
- CN115191352A CN115191352A CN202210735588.XA CN202210735588A CN115191352A CN 115191352 A CN115191352 A CN 115191352A CN 202210735588 A CN202210735588 A CN 202210735588A CN 115191352 A CN115191352 A CN 115191352A
- Authority
- CN
- China
- Prior art keywords
- hydroponic
- culture medium
- culture
- culturing
- medium
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 241000421576 Iris sp. ser. Hexagonae Species 0.000 title claims abstract description 21
- 238000002360 preparation method Methods 0.000 title claims abstract description 13
- 239000001963 growth medium Substances 0.000 claims abstract description 34
- 238000012258 culturing Methods 0.000 claims abstract description 23
- 230000001954 sterilising effect Effects 0.000 claims abstract description 21
- 241000196324 Embryophyta Species 0.000 claims abstract description 20
- 230000006698 induction Effects 0.000 claims abstract description 19
- 238000005406 washing Methods 0.000 claims abstract description 19
- 238000007605 air drying Methods 0.000 claims abstract description 13
- 238000004140 cleaning Methods 0.000 claims abstract description 13
- 238000004659 sterilization and disinfection Methods 0.000 claims abstract description 13
- 238000009630 liquid culture Methods 0.000 claims abstract description 12
- 238000000034 method Methods 0.000 claims abstract description 11
- 230000035755 proliferation Effects 0.000 claims abstract description 11
- 238000001035 drying Methods 0.000 claims abstract description 10
- 230000032683 aging Effects 0.000 claims abstract description 8
- 230000001939 inductive effect Effects 0.000 claims abstract description 4
- 238000007789 sealing Methods 0.000 claims abstract description 4
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims description 20
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 claims description 20
- 239000002609 medium Substances 0.000 claims description 17
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 16
- 238000005286 illumination Methods 0.000 claims description 12
- 229920001817 Agar Polymers 0.000 claims description 10
- 239000004471 Glycine Substances 0.000 claims description 10
- NWBJYWHLCVSVIJ-UHFFFAOYSA-N N-benzyladenine Chemical compound N=1C=NC=2NC=NC=2C=1NCC1=CC=CC=C1 NWBJYWHLCVSVIJ-UHFFFAOYSA-N 0.000 claims description 10
- 229930006000 Sucrose Natural products 0.000 claims description 10
- 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 description 10
- 239000008272 agar Substances 0.000 claims description 10
- 235000001968 nicotinic acid Nutrition 0.000 claims description 10
- 229960003512 nicotinic acid Drugs 0.000 claims description 10
- 239000011664 nicotinic acid Substances 0.000 claims description 10
- ZUFQODAHGAHPFQ-UHFFFAOYSA-N pyridoxine hydrochloride Chemical compound Cl.CC1=NC=C(CO)C(CO)=C1O ZUFQODAHGAHPFQ-UHFFFAOYSA-N 0.000 claims description 10
- 229960004172 pyridoxine hydrochloride Drugs 0.000 claims description 10
- 235000019171 pyridoxine hydrochloride Nutrition 0.000 claims description 10
- 239000011764 pyridoxine hydrochloride Substances 0.000 claims description 10
- 239000002994 raw material Substances 0.000 claims description 10
- 239000005720 sucrose Substances 0.000 claims description 10
- 229960000344 thiamine hydrochloride Drugs 0.000 claims description 10
- 235000019190 thiamine hydrochloride Nutrition 0.000 claims description 10
- 239000011747 thiamine hydrochloride Substances 0.000 claims description 10
- 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 description 10
- 229940064880 inositol 100 mg Drugs 0.000 claims description 9
- 230000002745 absorbent Effects 0.000 claims description 6
- 239000002250 absorbent Substances 0.000 claims description 6
- 239000008223 sterile water Substances 0.000 claims description 6
- 229920001213 Polysorbate 20 Polymers 0.000 claims description 4
- 229960002523 mercuric chloride Drugs 0.000 claims description 4
- LWJROJCJINYWOX-UHFFFAOYSA-L mercury dichloride Chemical compound Cl[Hg]Cl LWJROJCJINYWOX-UHFFFAOYSA-L 0.000 claims description 4
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 claims description 4
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 claims description 4
- 239000000126 substance Substances 0.000 claims description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 3
- OMOVVBIIQSXZSZ-UHFFFAOYSA-N [6-(4-acetyloxy-5,9a-dimethyl-2,7-dioxo-4,5a,6,9-tetrahydro-3h-pyrano[3,4-b]oxepin-5-yl)-5-formyloxy-3-(furan-3-yl)-3a-methyl-7-methylidene-1a,2,3,4,5,6-hexahydroindeno[1,7a-b]oxiren-4-yl] 2-hydroxy-3-methylpentanoate Chemical compound CC12C(OC(=O)C(O)C(C)CC)C(OC=O)C(C3(C)C(CC(=O)OC4(C)COC(=O)CC43)OC(C)=O)C(=C)C32OC3CC1C=1C=COC=1 OMOVVBIIQSXZSZ-UHFFFAOYSA-N 0.000 claims description 3
- 230000000249 desinfective effect Effects 0.000 claims description 3
- 239000003599 detergent Substances 0.000 claims description 3
- 238000009499 grossing Methods 0.000 claims description 3
- 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 1
- 229960000367 inositol Drugs 0.000 claims 1
- 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 1
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 claims 1
- 239000000463 material Substances 0.000 abstract description 4
- 238000004161 plant tissue culture Methods 0.000 abstract description 2
- 238000002474 experimental method Methods 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 235000015210 Fockea angustifolia Nutrition 0.000 description 1
- 244000186654 Fockea angustifolia Species 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 229910001385 heavy metal Inorganic materials 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000010865 sewage Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 230000004083 survival effect 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
- A01H4/002—Culture media 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
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/20—Reduction of greenhouse gas [GHG] emissions in agriculture, e.g. CO2
- Y02P60/21—Dinitrogen oxide [N2O], e.g. using aquaponics, hydroponics or efficiency measures
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses a preparation method of Louisiana iris hydroponic aseptic seedlings. Relates to the technical field of plant tissue culture. Provides a culture medium and a preparation method of the aseptic seedling: collecting plants, cleaning, removing leaves and roots at the stem base part to enable the transverse stem to be smooth, and drying; sterilizing the surface, washing and sterilizing under the aseptic condition; sterilizing the surface, sealing, ultrasonically vibrating and cleaning, washing and air drying to obtain a bud body; cutting off redundant petiole and stem tissues, and inserting the cut tissue into a sterilization induction culture medium for culture; after the buds grow out, taking out the explant, cutting off the blackening and ageing part, and transferring the explant into a sterilization and proliferation culture medium for culture to obtain a transferred seedling; culturing to obtain complete hydroponic plant, cutting off old leaves and black rot tissues, and culturing in liquid culture medium; culturing the hydroponic plants and inducing root systems. The method can obtain aseptic seedlings with larger population quantity in a shorter time, the roots are hydroponic roots, and the method can be used for preparing experimental materials, and is strong in reproducibility and easy to operate.
Description
Technical Field
The invention relates to the technical field of plant tissue culture, in particular to a preparation method of Louisiana iris hydroponic aseptic seedlings.
Background
Louisiana Iris (Iris hybrids 'Louisiana') has stronger resistance and wider adaptability compared with dry-grown Iris, and the wet-grown Iris planted in the water body can degrade or adsorb heavy metals in the polluted water body, thereby having a certain repairing effect on eutrophic sewage. The research on the water-cultured aseptic Louisiana iris can be used as a scientific research material on one hand, the research on the action mechanism of the Louisiana iris in the aspect of water body purification on the other hand, and because the interference of environmental factors such as microorganisms exists during the wet cultivation and the induction of water roots has certain difficulty, the two problems can be well solved by the water-cultured aseptic seedling; on the other hand, the Louisiana iris has good ornamental value and practicability.
But the seedling propagation speed is low, the yield is low, a large number of plants can be obtained in a short time through tissue culture and rapid propagation, and the water culture seedling has strong adaptability to the environment and can be applied to water body restoration engineering. However, the existing culture media are solid culture media, the root system induced in the rooting culture stage is not suitable for water culture, and if a water culture experiment is needed subsequently for bottle seedlings, an aquatic root system still needs to be induced, so that the risk of pollution is increased, and the survival rate of the tissue culture seedlings is greatly reduced.
Therefore, the problem that needs to be solved by the technical personnel in the field is how to provide a preparation method of the Louisiana iris hydroponic aseptic seedling.
Disclosure of Invention
In view of this, the invention provides a preparation method of a Louisiana iris hydroponic aseptic seedling. The obtained aseptic seedlings are used as important materials for scientific research, and can effectively remove the interference of microorganisms on the surface of plants on experiments.
In order to achieve the purpose, the invention adopts the following technical scheme:
a culture medium for water culture of Louisiana iris aseptic seedlings comprises a liquid culture medium and liquid culture raw materials: NH 4 NO 3 825 mg/L、KNO 3 950 mg/L、MgSO 4 7H 2 O 185mg/L、KH 2 PO 4 85mg/L、CaCl 2 2H 2 O 220mg/L、KI 0.83mg/L、H 3 BO 3 6.2 mg/L、MnSO 4 4H 2 O 22.3mg/L、ZnSO 4 7H 2 O 8.6mg/L、Na 2 MoO 4 2H 2 O 0.25mg/L、CuSO 4 5H 2 O 0.025mg/L、CoCl 2 6H 2 O 0.025mg/L、FeSO 4 7H 2 O 13.9mg/L、Na 2 -EDTA 2H 2 O37.3 mg/L, inositol 100mg/L, glycine 2mg/L, thiamine hydrochloride 0.1mg/L, pyridoxine hydrochloride 0.5mg/L, nicotinic acid 0.5mg/L, 6-benzylaminopurine 0.05mg/L, NAA 0.5mg/L, and,1% of sucrose and 0.65% of agar; the pH was adjusted to 5.8.
Preferably, the following components: also comprises an induction culture medium and a multiplication culture medium which are used in combination; raw materials of the induction medium: NH 4 NO 3 825mg/L、KNO 3 950mg/L、MgSO 4 7H 2 O 185mg/L、KH 2 PO 4 85mg/L、CaCl 2 2H 2 O 220mg/L、KI 0.83mg/L、H 3 BO 3 6.2 mg/L、MnSO 4 4H 2 O 22.3mg/L、ZnSO 4 7H 2 O 8.6mg/L、Na 2 MoO 4 2H 2 O 0.25mg/L、CuSO 4 5H 2 O 0.025mg/L、CoCl 2 6H 2 O 0.025mg/L、FeSO 4 7 H 2 O 13.9mg/L、Na 2 -EDTA 2H 2 O37.3 mg/L, inositol 100mg/L, glycine 2mg/L, thiamine hydrochloride 0.1mg/L, pyridoxine hydrochloride 0.5mg/L, nicotinic acid 0.5mg/L, 6-benzylaminopurine 3mg/L, NAA 0.1mg/L, KT 2mg/L, 3% sucrose, 0.65% agar based on the total mass of the induction medium; adjusting the pH value to 5.8;
raw materials of the proliferation medium: NH (NH) 4 NO 3 1700 mg/L、KNO 3 1900 mg/L、MgSO 4 7H 2 O 370mg/L、KH2PO 4 170 mg/L、CaCl 2 2H 2 O 440mg/L、KI 0.83mg/L、H 3 BO 3 6.2mg/L、MnSO 4 4H 2 O 22.3mg/L、ZnSO 4 7H 2 O 8.6mg/L、Na 2 MoO 4 2H 2 O 0.25mg/L、CuSO 4 5H 2 O 0.025mg/L、CoCl 2 6H 2 O 0.025mg/L、FeSO 4 7H 2 O 27.8mg/L、Na 2 -EDTA 2H 2 O37.3 mg/L, inositol 100mg/L, glycine 2mg/L, thiamine hydrochloride 0.1mg/L, pyridoxine hydrochloride 0.5mg/L, nicotinic acid 0.5mg/L, 6-benzylaminopurine 2mg/L, NAA 0.1mg/L, KT 2mg/L, 3% sucrose, 0.65% agar based on the total mass of the induction medium, and adjusting pH to 5.8.
The invention also provides a preparation method of the Louisiana iris hydroponic aseptic seedling based on the culture medium, which comprises the following steps:
(1) Collecting Louisiana iris plants, cleaning, removing leaves and roots at stem base parts to enable transverse stems to be smooth, and drying to obtain an object A;
(2) Sterilizing the surface of the substance A, washing and sterilizing under the aseptic condition to obtain a substance B;
(3) Sterilizing the surface of the object B, sealing, ultrasonically vibrating and cleaning, washing and air drying to obtain a bud body;
(4) Cutting off redundant petiole and stem tissues from the bud body, reserving the bud body at the middle upper part, and inserting the bud body into a sterilization induction culture medium for culture;
(5) After the buds grow out, taking out the explants, cutting off melanized aging parts, and transferring the explants to a sterilization proliferation culture medium for culture to obtain transferred seedlings;
(6) Culturing the transferred seedlings to complete hydroponic plants, cutting off old leaves and black rot tissues, and transferring the seedlings to a liquid culture medium for culture;
(7) Culturing the hydroponic plants and inducing root systems.
Further, the Louisiana iris plant standard collected in the step (1): healthy and sturdy; no plant diseases and insect pests;
the steps (3) to (7) are all required to maintain an aseptic environment;
only the bud with the largest middle part is reserved in the step (4); the length is 1cm;
tool for cutting off blackened and aged part: a scalpel;
and (5) repeating the step (5), realizing the proliferation of the plants, and entering the next step after a certain number of proliferated seedlings are obtained.
The equipment used was: an ultra-clean workbench.
Preferably: step (1) basal stem portion: 2-3 cm away from the stem base; smoothing the transverse stem: the nodes that cross the surface of the stem are removed.
Preferably: the step (2) is specifically as follows: washing the step A with a detergent for 1-2 times, air-drying, and then disinfecting the surface of the step A with 75% alcohol for 1-1.5 min under the aseptic condition: washing with sterile water, and drying with sterilized absorbent paper.
Preferably: step (3) disinfection: 0.1% mercuric chloride, adding Tween-20 before use; and (3) disinfection time: 5min; ultrasonic vibration cleaning for 3min; washing and air drying: washing with sterile water for 3 times, drying with sterilized absorbent paper, and air drying.
Further, the dosage ratio of mercuric chloride to tween-20 is as follows: 100ml:4 drops, can be adjusted by proper fine adjustment;
ultrasonic: 40KH or other conventional parameters.
Preferably: and (4) culturing environment: controlling the temperature at 28 ℃, culturing for 12h in the dark, and transferring to the environment with the illumination intensity of 5000lux and the illumination duration of 12h for culturing.
Preferably: step (5), cutting off the blackening and ageing part by using a scalpel, and transferring the blackening and ageing part into a sterilized proliferation culture medium; the temperature of the culture environment is controlled at 28 ℃, the illumination intensity is 5000lux, and the illumination time is 12h.
Preferably: and (5) culturing the whole plant in the step (6) for 30d.
Preferably: and (7) culturing until: the plants grew to 8cm.
According to the technical scheme, compared with the prior art, the preparation method of the Louisiana iris hydroponic aseptic seedlings has the advantages that the aseptic seedlings with larger population quantity can be obtained in a shorter time, the roots are hydroponic roots, the method can be used for preparing experimental materials, and the method is high in reproducibility and easy to operate.
Detailed Description
The following will clearly and completely describe the technical solutions in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The embodiment of the invention discloses a preparation method of Louisiana iris hydroponic aseptic seedlings.
Example 1
A culture medium for water culture of Louisiana iris aseptic seedlings comprises a liquid culture medium and liquid culture raw materials: NH (NH) 4 NO 3 825 mg/L、KNO 3 950 mg/L、MgSO 4 7H 2 O 185mg/L、KH 2 PO 4 85mg/L、CaCl 2 2H 2 O 220mg/L、KI 0.83mg/L、H 3 BO 3 6.2 mg/L、MnSO 4 4H 2 O 22.3mg/L、ZnSO 4 7H 2 O 8.6mg/L、Na 2 MoO 4 2H 2 O 0.25mg/L、CuSO 4 5H 2 O 0.025mg/L、CoCl 2 6H 2 O 0.025mg/L、FeSO 4 7H 2 O 13.9mg/L、Na 2 -EDTA 2H 2 O37.3 mg/L, inositol 100mg/L, glycine 2mg/L, thiamine hydrochloride 0.1mg/L, pyridoxine hydrochloride 0.5mg/L, nicotinic acid 0.5mg/L, 6-benzylaminopurine 0.05mg/L, NAA 0.5mg/L, sucrose 1%, agar 0.65%; the pH was adjusted to 5.8.
Example 2
A culture medium for water culture of Louisiana iris aseptic seedlings comprises a liquid culture medium and liquid culture raw materials: NH (NH) 4 NO 3 825 mg/L、KNO 3 950 mg/L、MgSO 4 7H 2 O 185mg/L、KH 2 PO 4 85mg/L、CaCl 2 2H 2 O 220mg/L、KI 0.83mg/L、H 3 BO 3 6.2 mg/L、MnSO 4 4H 2 O 22.3mg/L、ZnSO 4 7H 2 O 8.6mg/L、Na 2 MoO 4 2H 2 O 0.25mg/L、CuSO 4 5H 2 O 0.025mg/L、CoCl 2 6H 2 O 0.025mg/L、FeSO 4 7H 2 O 13.9mg/L、Na 2 -EDTA 2H 2 O37.3 mg/L, inositol 100mg/L, glycine 2mg/L, thiamine hydrochloride 0.1mg/L, pyridoxine hydrochloride 0.5mg/L, nicotinic acid 0.5mg/L, based on the total mass of the induction medium, 6-benzylaminopurine 0.05mg/L, NAA 0.5mg/L, 1% sucrose, 0.65% agar; the pH was adjusted to 5.8.
Also comprises an induction culture medium and a multiplication culture medium which are used in combination; raw materials of the induction medium: NH (NH) 4 NO 3 825mg/L、KNO 3 950mg/L、MgSO 4 7H 2 O 185mg/L、KH 2 PO 4 85 mg/L、CaCl 2 2H 2 O 220mg/L、KI 0.83mg/L、H 3 BO 3 6.2 mg/L、MnSO 4 4H 2 O 22.3mg/L、ZnSO 4 7H 2 O 8.6mg/L、Na 2 MoO 4 2H 2 O 0.25mg/L、CuSO 4 5H 2 O 0.025mg/L、CoCl 2 6H 2 O 0.025mg/L、FeSO 4 7H 2 O 13.9mg/L、Na 2 -EDTA 2H 2 O37.3 mg/L, inositol 100mg/L, glycine 2mg/L, thiamine hydrochloride 0.1mg/L, pyridoxine hydrochloride 0.5mg/L, nicotinic acid 0.5mg/L, 6-benzylaminopurine 3mg/L, NAA 0.1mg/L, KT 2mg/L, 3% sucrose, 0.65% agar based on the total mass of the induction medium; adjusting the pH value to 5.8;
raw materials of the proliferation medium: NH 4 NO 3 1700 mg/L、KNO 3 1900 mg/L、MgSO 4 7H 2 O 370mg/L、KH 2 PO 4 170 mg/L、CaCl 2 2H 2 O 440mg/L、KI 0.83mg/L、H 3 BO 3 6.2mg/L、MnSO 4 4H 2 O 22.3mg/L、ZnSO 4 7H 2 O 8.6mg/L、Na 2 MoO 4 2H 2 O 0.25mg/L、CuSO 4 5H 2 O 0.025mg/L、CoCl 2 6H 2 O 0.025mg/L、FeSO 4 7H 2 O 27.8mg/L、Na 2 -EDTA 2H 2 O37.3 mg/L, inositol 100mg/L, glycine 2mg/L, thiamine hydrochloride 0.1mg/L, pyridoxine hydrochloride 0.5mg/L, nicotinic acid 0.5mg/L, 6-benzylaminopurine 2mg/L, NAA 0.1mg/L, KT 2mg/L, 3% sucrose, 0.65% agar based on the total mass of the induction medium, and adjusting pH to 5.8.
Example 3
A preparation method of Louisiana iris hydroponic aseptic seedlings based on the culture medium comprises the following steps:
(1) Collecting Louisiana iris plants, cleaning, removing leaves and roots at stem base parts to enable transverse stems to be smooth, and drying to obtain an object A;
(2) Sterilizing the surface of the substance A, washing and sterilizing under the aseptic condition to obtain a substance B;
(3) Sterilizing the surface of the object B, sealing, ultrasonically vibrating and cleaning, washing and air drying to obtain a bud body;
(4) Cutting off redundant petiole and stem tissues of the bud body, reserving the bud body at the middle upper part, and inserting the bud body into a sterilization induction culture medium for culture;
(5) After the buds grow out, taking out the explant, cutting off the blackening and ageing part, and transferring the explant into a sterilization and proliferation culture medium for culture to obtain a transferred seedling;
(6) Culturing the transferred seedlings to complete hydroponic plants, cutting off old leaves and black rot tissues, and transferring the seedlings to a liquid culture medium for culture;
(7) Culturing the hydroponic plants and inducing root systems.
In order to further optimize the technical scheme: step (1) stem base: 3-4 cm away from the stem base; smoothing the transverse stem: the nodes that cross the surface of the stem are removed. This example is 3cm.
In order to further optimize the technical scheme: the step (2) is specifically as follows: washing the step A with a detergent for 1-2 times, air-drying, and then disinfecting the surface of the step A with 75% alcohol for 1-1.5 min under the aseptic condition: washing with sterile water, and drying with sterilized absorbent paper.
In order to further optimize the technical scheme: step (3) disinfection: 0.1% mercuric chloride, adding Tween-20 before use; and (3) disinfection time: 5min; ultrasonic vibration cleaning for 3min; washing and air drying: washing with sterile water for 3 times, drying with sterilized absorbent paper, and air drying.
In order to further optimize the technical scheme: and (4) culturing environment: controlling the temperature at 28 ℃, culturing for 12h in the dark, and transferring to the environment with the illumination intensity of 5000lux and the illumination duration of 12h for culturing.
In order to further optimize the technical scheme: step (5), cutting off the blackening and ageing part by using a scalpel, and transferring the blackening and ageing part into a sterilized proliferation culture medium; the temperature of the culture environment is controlled at 28 ℃, the illumination intensity is 5000lux, and the illumination time is 12h.
In order to further optimize the technical scheme: and (5) culturing the whole plant in the step (6) for 30d.
In order to further optimize the technical scheme: and (7) culturing until: the plants grew to 8cm.
The embodiments in the present description are described in a progressive manner, each embodiment focuses on differences from other embodiments, and the same and similar parts among the embodiments are referred to each other.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (10)
1. A culture medium of Louisiana iris hydroponic aseptic seedlings is characterized by comprising a liquid culture medium, wherein the liquid culture medium comprises the following raw materials: NH (NH) 4 NO 3 825mg/L、KNO 3 950mg/L、MgSO 4 7H 2 O 185mg/L、KH 2 PO 4 85mg/L、CaCl 2 2H 2 O 220mg/L、KI 0.83mg/L、H 3 BO 3 6.2mg/L、MnSO 4 4H 2 O 22.3mg/L、ZnSO 4 7H 2 O 8.6mg/L、Na 2 MoO 4 2H 2 O 0.25mg/L、CuSO 4 5H 2 O 0.025mg/L、CoCl 2 6H 2 O 0.025mg/L、FeSO 4 7H 2 O 13.9mg/L、Na 2 -EDTA 2H 2 O37.3 mg/L, inositol 100mg/L, glycine 2mg/L, thiamine hydrochloride 0.1mg/L, pyridoxine hydrochloride 0.5mg/L, nicotinic acid 0.5mg/L, based on the total mass of the induction medium, 6-benzylaminopurine 0.05mg/L, NAA 0.5mg/L, 1% sucrose, 0.65% agar; the pH was adjusted to 5.8.
2. The culture medium of a hydroponic iris of claim 1 further comprising an induction medium and a multiplication medium used in combination; raw materials of the induction medium: NH 4 NO 3 825mg/L、KNO 3 950mg/L、MgSO 4 7H 2 O 185mg/L、KH 2 PO 4 85mg/L、CaCl 2 2H 2 O 220mg/L、KI 0.83mg/L、H 3 BO 3 6.2mg/L、MnSO 4 4H 2 O 22.3mg/L、ZnSO 4 7H 2 O 8.6mg/L、Na 2 MoO 4 2H 2 O 0.25mg/L、CuSO 4 5H 2 O 0.025mg/L、CoCl 2 6H 2 O 0.025mg/L、FeSO 4 7H 2 O 13.9mg/L、Na 2 -EDTA 2H 2 37.3mg/L of O, 100mg/L of inositol, 2mg/L of glycine, 0.1mg/L of thiamine hydrochloride, 0.5mg/L of pyridoxine hydrochloride, 0.5mg/L of nicotinic acid, 3mg/L of 6-benzylaminopurine, 0.1mg/L of NAA and 2mg/L of KT, 3 percent of sucrose and 0.65 percent of agar based on the total mass of the induction medium; adjusting the pH value to 5.8;
raw materials of the proliferation culture medium: NH 4 NO 3 1700mg/L、KNO 3 1900mg/L、MgSO 4 7H 2 O 370mg/L、KH 2 PO 4 170mg/L、CaCl 2 2H 2 O 440mg/L、KI 0.83mg/L、H 3 BO 3 6.2mg/L、MnSO 4 4H 2 O 22.3mg/L、ZnSO 4 7H 2 O 8.6mg/L、Na 2 MoO 4 2H 2 O 0.25mg/L、CuSO 4 5H 2 O 0.025mg/L、CoCl 2 6H 2 O 0.025mg/L、FeSO 4 7H 2 O 27.8mg/L、Na 2 -EDTA 2H 2 O37.3 mg/L, inositol 100mg/L, glycine 2mg/L, thiamine hydrochloride 0.1mg/L, pyridoxine hydrochloride 0.5mg/L, nicotinic acid 0.5mg/L, 6-benzylaminopurine 2mg/L, NAA 0.1mg/L, KT 2mg/L, 3% sucrose, 0.65% agar based on the total mass of the induction medium, and adjusting pH to 5.8.
3. A process for the preparation of hydroponic aseptic seedlings of iris venosa based on the medium of claim 1 or 2, comprising the steps of:
(1) Collecting Louisiana iris plants, cleaning, removing leaves and roots from the base of the iris plants to ensure smooth stem crossing, and drying to obtain a substance A;
(2) Sterilizing the surface of the substance A, washing and sterilizing under the aseptic condition to obtain a substance B;
(3) Sterilizing the surface of the object B, sealing, ultrasonically vibrating and cleaning, washing and air drying to obtain a bud body;
(4) Cutting off redundant petiole and stem tissues of the bud body, reserving the bud body at the middle upper part, and inserting the bud body into a sterilization induction culture medium for culture;
(5) After the buds grow out, taking out the explants, cutting off melanized aging parts, and transferring the explants to a sterilization proliferation culture medium for culture to obtain transferred seedlings;
(6) Culturing the transferred seedlings to complete hydroponic plants, cutting off old leaves and black rot tissues, and transferring the seedlings to a liquid culture medium for culture;
(7) Culturing the hydroponic plants and inducing root systems.
4. The method of claim 3, wherein the stem base of step (1): 2-3 cm away from the stem base; smoothing the transverse stem: the nodes that cross the surface of the stem are removed.
5. The preparation method according to claim 4, wherein the step (2) is specifically: cleaning the step A with a detergent for 1-2 times, air-drying, and then disinfecting the surface with 75% alcohol for 1-1.5 min under the aseptic condition: washing with sterile water, and drying with sterilized absorbent paper.
6. The method of claim 5, wherein the sterilizing of step (3): 0.1% mercuric chloride, and adding tween-20 before use; and (3) disinfection time: 5min; the ultrasonic vibration cleaning is carried out, and the cleaning time is 3min; washing and air drying: washing with sterile water for 3 times, drying with sterilized absorbent paper, and air drying.
7. The method of claim 6, wherein the culture environment of step (4): controlling the temperature at 28 ℃, culturing for 12h in the dark, and transferring to the environment with the illumination intensity of 5000lux and the illumination duration of 12h for culturing.
8. The method according to claim 7, wherein the melanized aged part is excised with a scalpel in step (5) and transferred to a sterilized proliferation medium; the temperature of the culture environment is controlled at 28 ℃, the illumination intensity is 5000lux, and the illumination time is 12h.
9. The method of claim 8, wherein the time period from the culturing in step (6) to the completion of the plant is 30 days.
10. The method of claim 8, wherein the culturing of step (7) is carried out until: the plants grew to 8cm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210735588.XA CN115191352A (en) | 2022-06-27 | 2022-06-27 | Preparation method of Louisiana iris hydroponic aseptic seedlings |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210735588.XA CN115191352A (en) | 2022-06-27 | 2022-06-27 | Preparation method of Louisiana iris hydroponic aseptic seedlings |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN115191352A true CN115191352A (en) | 2022-10-18 |
Family
ID=83579001
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210735588.XA Pending CN115191352A (en) | 2022-06-27 | 2022-06-27 | Preparation method of Louisiana iris hydroponic aseptic seedlings |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115191352A (en) |
-
2022
- 2022-06-27 CN CN202210735588.XA patent/CN115191352A/en active Pending
Non-Patent Citations (2)
| Title |
|---|
| 朱旭东等: "路易斯安娜鸢尾组培和苗期生长规律初步研究", 《福建林业科技》 * |
| 马元山等: "鸢尾的水培技术研究", 《中国农学通报》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101849506A (en) | Tissue culture and rapid propagation method of acer palmatum | |
| CN106718892A (en) | A kind of Chinese rose rapid propagation method | |
| CN104488716A (en) | Method for tissue culture of water nymph | |
| CN108040882A (en) | A kind of abductive approach of garlic test tube seedling | |
| CN109452170B (en) | Method for callus culture induced by cordyceps sobolifera roots | |
| GRUSELLE et al. | Walnut micropropagation | |
| CN101485290A (en) | Method for establishing rubber tree internal integument regeneration system | |
| CN103004588B (en) | Tissue culture and rapid propagation method of tulipa edulis | |
| CN110089427A (en) | A kind of in-vitro conservation method of bud germ plasm resource | |
| CN104604689B (en) | The method of quick acquisition witloof explant and its healing rate of raising | |
| CN102893866B (en) | Strawberry root tip detoxification and tissue culture method | |
| WO2021189544A1 (en) | Method for efficiently detoxifying lilies | |
| CN109287490B (en) | Method for rapidly propagating high-quality seedling of aquatic plant water soft-shelled turtle and application | |
| CN109984039B (en) | Lycoris radiata tissue culture method | |
| CN115191352A (en) | Preparation method of Louisiana iris hydroponic aseptic seedlings | |
| CN104026010B (en) | A kind of abductive approach of mulberry tree cotyledon indefinite bud | |
| CN103798138B (en) | A kind of is the method that outer implant sets up Bulbus Lilii embryo callus subculture regenerating system with gynoecium | |
| CN103798139B (en) | A kind of is the method that outer implant sets up Bulbus Lilii embryo callus subculture regenerating system with petal | |
| CN114532227B (en) | Method for inducing and proliferating calluses of agapanthus radicis roots and tips | |
| CN114600772A (en) | Tissue culture method and rapid propagation method of michelia figo in remote mountains | |
| Faggioli et al. | In vitro micrografting of Pyrus communis shoot tips | |
| CN109548655B (en) | Tissue culture method of quanlang tree | |
| CN107135943A (en) | A kind of winter cherry rapid propagation in vitro method | |
| CN109479727B (en) | Method for inducing embryonic cells by taking agapanthus praecox leaves as explants | |
| JPH02222626A (en) | Crossbred plant of genus allium and breeding of the crossbred plant |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20221018 |