CN110999792A - Multi-bud culture method for anoectochilus roxburghii - Google Patents
Multi-bud culture method for anoectochilus roxburghii Download PDFInfo
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- CN110999792A CN110999792A CN201911391856.5A CN201911391856A CN110999792A CN 110999792 A CN110999792 A CN 110999792A CN 201911391856 A CN201911391856 A CN 201911391856A CN 110999792 A CN110999792 A CN 110999792A
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- 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
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- 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
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Abstract
The invention discloses a method for culturing multiple buds of anoectochilus formosanus, which comprises the following steps: the method comprises the following steps of collecting explants of wild anoectochilus formosanus or seeds of the anoectochilus formosanus, and carrying out disinfection treatment, wherein the disinfection treatment method comprises the following steps: sterilizing with 75% alcohol, sequentially placing into seeding culture medium, proliferation culture medium, strong seedling culture medium and rooting culture medium, culturing in 45 days culture room, and transplanting. By adopting the culture method, two or more new buds can be generated by inducing one bud-containing stem segment, the production rate can be improved, the production cost can be reduced, the growth time of a tissue culture room can be shortened, and the market competitiveness of a company can be improved.
Description
Technical Field
The invention relates to a method for culturing multiple buds of anoectochilus formosanus, in particular to the technical field of anoectochilus formosanus culture.
Background
Anoectochilus roxburghii (also known as Anoectochilus roxburghii) is a plant of the genus Labiatae. In folks, there are known the names of gold thread, golden earring, bird ginseng, and gold thread, such as Guxiao, jin Qian Cao, and Jinxiao Shi Song. Anoectochilus roxburghii herb is sweet and mild in taste when used as a medicine. Anoectochilus roxburghii has effects of clearing heat and cooling blood, dispelling pathogenic wind and promoting diuresis, removing toxic substance, relieving pain, and relieving cough. At present, the existing anoectochilus formosanus adopts artificial culture, the culture survival rate is low, and in order to increase the survival rate, a multibud culture method of the anoectochilus formosanus is researched.
Disclosure of Invention
The invention aims to provide a method for culturing multiple buds of anoectochilus formosanus.
In order to achieve the purpose, the invention provides the following technical scheme: a method for culturing multiple buds of anoectochilus formosanus comprises the following steps: the method comprises the following steps of collecting explants of wild anoectochilus formosanus or seeds of the anoectochilus formosanus, and carrying out disinfection treatment, wherein the disinfection treatment method comprises the following steps: sterilizing with 75% alcohol, sequentially placing into seeding culture medium, proliferation culture medium, strong seedling culture medium and rooting culture medium, culturing in 45 days culture room, and transplanting.
Preferably, the seeding culture medium in the culture process comprises the following raw materials: liquid A: 1000 ml; b, liquid: 1000 ml; c, liquid: 100 ml of water; d, liquid: 100 ml of water; l liquid: 20 ml of the solution; m1 liquid: 8 ml of water; sucrose: 400 g; agar: 76g of a carrier; carbon powder: 36g of a mixture; potato: 2000 g; adding sodium hydroxide when the culture medium turns yellow; adding hydrochloric acid when blackening, wherein the pH value is as follows: 5.4-5.8;
the proliferation medium comprises the following raw materials: liquid A: 1000 ml; b, liquid: 1000 ml; c, liquid: 100 ml of water; d, liquid: 100 ml of water; l liquid: 20 ml of the solution; m1 liquid: 8 ml, M2 liquid: 5 ml of water; sucrose: 400 g; agar: 76g of a carrier; carbon powder: 36g of a mixture; potato: 2000 g; adding sodium hydroxide when the culture medium turns yellow; adding hydrochloric acid when blackening; the pH value is: 5.4-5.8;
the strong seedling culture medium comprises the following raw materials: liquid A: 1000 ml; b, liquid: 1000 ml; c, liquid: 100 ml of water; d, liquid: 100 ml of water; l liquid: 20 ml of the solution; m1 liquid: 15 ml, M2 liquid: 8 ml of water; sucrose: 400 g; agar: 76g of a carrier; carbon powder: 40g of the total weight of the mixture; potato: 2000 g; adding sodium hydroxide when the culture medium turns yellow; adding hydrochloric acid when blackening; the pH value is: 5.4-5.8;
the rooting medium comprises the following raw materials: 1000 ml of liquid A; b, liquid: 1000 ml; c, liquid: 100 ml of water; d, liquid: 100 ml of water; l liquid: 20 ml of the solution; m1 liquid: 20 ml, M2 liquid: 10 ml of water; sucrose: 500, a step of; 76g of agar; carbon powder: 40g of the total weight of the mixture; banana: 2500 g; adding sodium hydroxide when the culture medium turns yellow; adding hydrochloric acid when blackening, wherein the pH value is as follows: 5.4-5.8;
preferably, the formula of the liquid A comprises the following raw materials: ammonium nitrate: 33.8 g; potassium nitrate: 33g of a mixture; magnesium sulfate: 4.9 g; 3.5g of monopotassium phosphate;
the formula of the liquid B comprises the following raw materials: water 9.4g, calcium chloride: 9.4 g; firstly, discharging water and then discharging calcium chloride;
the formula of the liquid C comprises the following raw materials: glycine: 0.8 g; vitamin B1: 0.05 g; nicotinic acid: 0.5 g; pyridoxine hydrochloride VB 6: 0.2g of the formulas are combined and dissolved into a whole; inositol: 25g were dissolved separately; then the two are combined and dissolved into a whole;
the formula of the liquid D comprises the following raw materials: ferrous sulfate: 6.39 g; disodium ethylene diamine tetraacetate: 9.78g of the two are respectively dissolved and then mixed;
the liquid formula L comprises the following raw materials: manganese sulfate: 16.9 g; zinc sulfate: 8.6 g; boric acid: 6.2 g; potassium iodide: 0.83 g; copper sulfate: 0.25 g; cobalt chloride: 0.25 g; mixing and dissolving the substances; sodium molybdate: 0.25 g; the material is dissolved separately, and the two are dissolved together;
m1 liquid formulation: a-naphthylacetic acid: 0.5 g; sodium hydroxide: 0.55g, and storing in a refrigerator for preservation;
m2 liquid formulation: 0.55g of 6-benzyladenine; 0.45g of sodium hydroxide, and storing in the dark.
Compared with the prior art, the invention has the following beneficial effects: by adopting the culture method, two or more new buds can be generated by inducing one bud-containing stem segment, the production rate can be improved, the production cost can be reduced, the growth time of a tissue culture room can be shortened, and the market competitiveness of a company can be improved.
Detailed Description
The technical solutions of the present invention will be described clearly and completely in the following embodiments of the present invention, and it should be understood 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.
Examples
A method for culturing multiple buds of anoectochilus formosanus comprises the following steps: the method comprises the following steps of collecting explants of wild anoectochilus formosanus or seeds of the anoectochilus formosanus, and carrying out disinfection treatment, wherein the disinfection treatment method comprises the following steps: sterilizing with 75% alcohol, sequentially placing into seeding culture medium, proliferation culture medium, strong seedling culture medium and rooting culture medium, culturing in 45 days culture room, and transplanting.
Wherein the seeding culture medium comprises the following raw materials in the culture process: liquid A: 1000 ml; b, liquid: 1000 ml; c, liquid: 100 ml of water; d, liquid: 100 ml of water; l liquid: 20 ml of the solution; m1 liquid: 8 ml of water; sucrose: 400 g; agar: 76g of a carrier; carbon powder: 36g of a mixture; potato: 2000 g; adding sodium hydroxide when the culture medium turns yellow; adding hydrochloric acid when blackening, wherein the pH value is as follows: 5.4-5.8;
the proliferation medium comprises the following raw materials: liquid A: 1000 ml; b, liquid: 1000 ml; c, liquid: 100 ml of water; d, liquid: 100 ml of water; l liquid: 20 ml of the solution; m1 liquid: 8 ml, M2 liquid: 5 ml of water; sucrose: 400 g; agar: 76g of a carrier; carbon powder: 36g of a mixture; potato: 2000 g; adding sodium hydroxide when the culture medium turns yellow; adding hydrochloric acid when blackening; the pH value is: 5.4-5.8;
the strong seedling culture medium comprises the following raw materials: liquid A: 1000 ml; b, liquid: 1000 ml; c, liquid: 100 ml of water; d, liquid: 100 ml of water; l liquid: 20 ml of the solution; m1 liquid: 15 ml, M2 liquid: 8 ml of water; sucrose: 400 g; agar: 76g of a carrier; carbon powder: 40g of the total weight of the mixture; potato: 2000 g; adding sodium hydroxide when the culture medium turns yellow; adding hydrochloric acid when blackening; the pH value is: 5.4-5.8;
the rooting medium comprises the following raw materials: 1000 ml of liquid A; b, liquid: 1000 ml; c, liquid: 100 ml of water; d, liquid: 100 ml of water; l liquid: 20 ml of the solution; m1 liquid: 20 ml, M2 liquid: 10 ml of water; sucrose: 500, a step of; 76g of agar; carbon powder: 40g of the total weight of the mixture; banana: 2500 g; adding sodium hydroxide when the culture medium turns yellow; adding hydrochloric acid when blackening, wherein the pH value is as follows: 5.4-5.8;
preferably, the formula of the liquid A comprises the following raw materials: ammonium nitrate: 33.8 g; potassium nitrate: 33g of a mixture; magnesium sulfate: 4.9 g; 3.5g of monopotassium phosphate;
the formula of the liquid B comprises the following raw materials: water 9.4g, calcium chloride: 9.4 g; firstly, discharging water and then discharging calcium chloride;
the formula of the liquid C comprises the following raw materials: glycine: 0.8 g; vitamin B1: 0.05 g; nicotinic acid: 0.5 g; pyridoxine hydrochloride VB 6: 0.2g of the formulas are combined and dissolved into a whole; inositol: 25g were dissolved separately; then the two are combined and dissolved into a whole;
the formula of the liquid D comprises the following raw materials: ferrous sulfate: 6.39 g; disodium ethylene diamine tetraacetate: 9.78g of the two are respectively dissolved and then mixed;
the liquid formula L comprises the following raw materials: manganese sulfate: 16.9 g; zinc sulfate: 8.6 g; boric acid: 6.2 g; potassium iodide: 0.83 g; copper sulfate: 0.25 g; cobalt chloride: 0.25 g; mixing and dissolving the substances; sodium molybdate: 0.25 g; the material is dissolved separately, and the two are dissolved together;
m1 liquid formulation: a-naphthylacetic acid: 0.5 g; sodium hydroxide: 0.55g, and storing in a refrigerator for preservation;
m2 liquid formulation: 0.55g of 6-benzyladenine; 0.45g of sodium hydroxide, and storing in the dark. Comparative example 1: a method for culturing multiple buds of anoectochilus formosanus comprises the following steps: the method comprises the following steps of collecting explants of wild anoectochilus formosanus or seeds of the anoectochilus formosanus, and carrying out disinfection treatment, wherein the disinfection treatment method comprises the following steps: the seed culture medium and the propagation culture medium in the above examples were sequentially placed in a 75% alcohol sterilization treatment, and grown into large seedlings in a 45-day culture room.
Comparative example 2: a high-quality anoectochilus formosanus tissue culture method comprises the following steps: the method comprises the following steps of collecting explants of wild anoectochilus formosanus or seeds of the anoectochilus formosanus, and carrying out disinfection treatment, wherein the disinfection treatment method comprises the following steps: sterilizing with 75% alcohol, sequentially placing into strong seedling culture medium and rooting culture medium in the above embodiments, and culturing in 45 days culture room to obtain large seedlings.
Comparative example 3: a high-quality anoectochilus formosanus tissue culture method comprises the following steps: the method comprises the following steps of collecting explants of wild anoectochilus formosanus or seeds of the anoectochilus formosanus, and carrying out disinfection treatment, wherein the disinfection treatment method comprises the following steps: sterilizing with 75% alcohol, and culturing in culture room for 45 days to obtain large seedlings.
The following table shows the conditions of culturing anoectochilus formosanus tissue seedlings in different examples and comparative examples:
| incubation time | Height cm of seedling | Survival rate of transplantation | |
| Examples | 45 days | 3.8 | 96.5％ |
| Comparative example 1 | 45 days | 3.0 | 78％ |
| Comparative example 2 | 45 days | 2.3 | 66％ |
| Comparative example 3 | 45 days | 2.2 | 54％ |
The experimental data show that the method for culturing the anoectochilus formosanus tissue greatly shortens the growth time of a tissue culture room and improves the survival rate of the anoectochilus formosanus tissue.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (3)
1. A method for culturing anoectochilus formosanus multiple buds is characterized by comprising the following steps: comprises the following steps: the method comprises the following steps of collecting explants of wild anoectochilus formosanus or seeds of the anoectochilus formosanus, and carrying out disinfection treatment, wherein the disinfection treatment method comprises the following steps: sterilizing with 75% alcohol, sequentially placing into seeding culture medium, proliferation culture medium, strong seedling culture medium and rooting culture medium, culturing in 45 days culture room, and transplanting.
2. The method for culturing multiple buds of Anoectochilus roxburghii according to claim 1, which is characterized in that: the seeding culture medium in the culture process comprises the following raw materials: liquid A: 1000 ml; b, liquid: 1000 ml; c, liquid: 100 ml of water; d, liquid: 100 ml of water; l liquid: 20 ml of the solution; m1 liquid: 8 ml of water; sucrose: 400 g; agar: 76g of a carrier; carbon powder: 36g of a mixture; potato: 2000 g; adding sodium hydroxide when the culture medium turns yellow; adding hydrochloric acid when blackening, wherein the pH value is as follows: 5.4-5.8;
the proliferation medium comprises the following raw materials: liquid A: 1000 ml; b, liquid: 1000 ml; c, liquid: 100 ml of water; d, liquid: 100 ml of water; l liquid: 20 ml of the solution; m1 liquid: 8 ml, M2 liquid: 5 ml of water; sucrose: 400 g; agar: 76g of a carrier; carbon powder: 36g of a mixture; potato: 2000 g; adding sodium hydroxide when the culture medium turns yellow; adding hydrochloric acid when blackening; the pH value is: 5.4-5.8;
the strong seedling culture medium comprises the following raw materials: liquid A: 1000 ml; b, liquid: 1000 ml; c, liquid: 100 ml of water; d, liquid: 100 ml of water; l liquid: 20 ml of the solution; m1 liquid: 15 ml, M2 liquid: 8 ml of water; sucrose: 400 g; agar: 76g of a carrier; carbon powder: 40g of the total weight of the mixture; potato: 2000 g; adding sodium hydroxide when the culture medium turns yellow; adding hydrochloric acid when blackening; the pH value is: 5.4-5.8;
the rooting medium comprises the following raw materials: 1000 ml of liquid A; b, liquid: 1000 ml; c, liquid: 100 ml of water; d, liquid: 100 ml of water; l liquid: 20 ml of the solution; m1 liquid: 20 ml, M2 liquid: 10 ml of water; sucrose: 500, a step of; 76g of agar; carbon powder: 40g of the total weight of the mixture; banana: 2500 g; adding sodium hydroxide when the culture medium turns yellow; adding hydrochloric acid when blackening, wherein the pH value is as follows: 5.4-5.8.
3. The method for culturing multiple buds of Anoectochilus roxburghii according to claim 1, which is characterized in that: the formula of the liquid A comprises the following raw materials: ammonium nitrate: 33.8 g; potassium nitrate: 33g of a mixture; magnesium sulfate: 4.9 g; 3.5g of monopotassium phosphate;
the formula of the liquid B comprises the following raw materials: water 9.4g, calcium chloride: 9.4 g; firstly, discharging water and then discharging calcium chloride;
the formula of the liquid C comprises the following raw materials: glycine: 0.8 g; vitamin B1: 0.05 g; nicotinic acid: 0.5 g; pyridoxine hydrochloride VB 6: 0.2g of the formulas are combined and dissolved into a whole; inositol: 25g were dissolved separately; then the two are combined and dissolved into a whole;
the formula of the liquid D comprises the following raw materials: ferrous sulfate: 6.39 g; disodium ethylene diamine tetraacetate: 9.78g of the two are respectively dissolved and then mixed;
the liquid formula L comprises the following raw materials: manganese sulfate: 16.9 g; zinc sulfate: 8.6 g; boric acid: 6.2 g; potassium iodide: 0.83 g; copper sulfate: 0.25 g; cobalt chloride: 0.25 g; mixing and dissolving the substances; sodium molybdate: 0.25 g; the material is dissolved separately, and the two are dissolved together;
m1 liquid formulation: a-naphthylacetic acid: 0.5 g; sodium hydroxide: 0.55g, and storing in a refrigerator for preservation;
m2 liquid formulation: 0.55g of 6-benzyladenine; 0.45g of sodium hydroxide, and storing in the dark.
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Application publication date: 20200414 |