CN109122206A - A kind of method for culturing seedlings of dendrobium candidum endogenetic fungus - Google Patents
A kind of method for culturing seedlings of dendrobium candidum endogenetic fungus Download PDFInfo
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- CN109122206A CN109122206A CN201811042090.5A CN201811042090A CN109122206A CN 109122206 A CN109122206 A CN 109122206A CN 201811042090 A CN201811042090 A CN 201811042090A CN 109122206 A CN109122206 A CN 109122206A
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- seed
- dendrobium candidum
- endogenetic fungus
- dendrobium
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/20—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material
- A01G24/28—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material containing peat, moss or sphagnum
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/20—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G31/00—Soilless cultivation, e.g. hydroponics
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
- C05F11/02—Other organic fertilisers from peat, brown coal, and similar vegetable deposits
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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Pretreatment Of Seeds And Plants (AREA)
- Cultivation Of Plants (AREA)
Abstract
The present invention provides a kind of method for culturing seedlings of dendrobium candidum, the method for culturing seedlings includes: (1) seed treatment, wins mature dendrobium candidum fruit pod and places ventilating and cooling place and dries；(2) matrix prepares, and sterilizing peat soil is initially charged in seed plate, is reloaded into the bacterial manure for being mixed with endogenetic fungus, flattens；(3) it sows, seed is fitted into seed dispenser, uniform broadcasting is on seed plate；(4) moisturizing is handled, and encases seed plate with preservative film, and punches several, the shady place placement under outdoor room temperature above.It is fast that breeding method of the invention sprouts Seeds of Dendrobium Candidum, at low cost.
Description
Technical field
The present invention relates to growing and cultivation field, in particular to a kind of method for culturing seedlings of Seeds of Dendrobium Candidum.
Background technique
Dendrobium candidum has a variety of medicinal efficacies, and stem is used as medicine, and belong to the yin tonics in tonic: reinforcing stomach reg fluid, enriching yin are clear
Heat.Ancient Chinese medicine doctor all thinks that " dendrobium candidum " has the function of nourishing hepatic yin, is the key medicine for treating various liver and bladder diseases, can be used for controlling
Treat the diseases in the liver and gallbladder such as hepatitis, cholecystitis, gall stone.
There is natural symbiosis, seed is very tiny for dendrobium candidum and mycorrhizal fungi, is the incomplete embryo of differentiation
Cell, it is internal to be free of endosperm, it cannot normally sprout under field conditions (factors), only the mycorhiza of fungi and seed or root formation symbiosis
After relationship, could normally it sprout and growth and development.Seeds of Dendrobium Candidum is sprouted at present and the conventional method of nursery is to pass through group
Training to complete, and set up a transfer room that can be run and tissue culture room be need many cost inputs to carry out purchase of equipment, and
Need to take a large amount of working hours to complete a series of working hours such as the sub-bottle cultivation after the production of culture medium, seed sprouting.Of the invention
Method is a kind of sprouting and nursery simple and quick, that can complete in normal environment seed, is eliminated with tissue culture method
Equipment and working hour greatly reduce the production cost of seedlings of Dendrobium officinale.
Summary of the invention
The purpose of the present invention is to provide a kind of breeding method of Seeds of Dendrobium Candidum, breeding method of the invention can be very
The sprouting and nursery of good promotion Seeds of Dendrobium Candidum, this can save a series of activities and its generation of cumbersome artificial tissue culture
Expense has significant application value.
To achieve the above object, the present invention adopts the following technical scheme:
The present invention cultivates Seeds of Dendrobium Candidum using dendrobium candidum endogenetic fungus bacterial manure, makes fungi and Seed Development symbiosis,
Without the sprouting and nursery of seed can be realized by tissue culture.
Specific step is as follows for the above method:
(1) seed is picked: after mature dendrobium candidum fruit pod picking, being gently rinsed with water fruit pod epidermis, is placed on clean white
It is dried on paper in shady place, until fruit pod cracks, collection seed is loaded in seed dispenser；
(2) prepared by matrix: seedling medium peat soil is packed at the 1/2-3/4 of seedlings nursing plate, then filled up with endogenetic fungus bacterial manure, it is whole
Flat, watering is wet to matrix；
(3) seed is sowed: gently shaking seeding device sows dendrobium nobile seed uniformly in stromal surface, and one layer of guarantor is covered after having sowed
Fresh film has moistening effect and beats several duck eyes on preservative film；
(4) post-broadcast management: after planting seed plate places the shady place under room temperature, and after planting the 5th day seed, which expands, turns green sprouting
(Fig. 1), keeping the water content of surface layer seedling medium is 30-35%, every 20-30 days spraying primary filtered bacterium solution liquid after germination
Body obtains nascent dendrobium nobile seedling；
(5) hardening: dendrobium nobile seedling to be come into being is long to 3 true leaves, carries out hardening to nascent dendrobium nobile seedling, continuous hardening 15 days obtains iron
Skin dendrobium nobile seedling (Fig. 2).
Bacterium solution liquid the preparation method is as follows: being chosen from Epulorhiza solid medium with transfer needle in the step (4)
Take 0.7cm2Epulorhiza strain, access in the PDA liquid medium of 400mL, 28 DEG C, 200rpm cultivate 10 days after, use
0.22 μm of membrane filtration removes thallus to get bacterium solution liquid is arrived.
The endogenetic fungus bacterial manure the preparation method is as follows:
(1) use transfer needle from picking 0.7cm on Epulorhiza solid medium2Epulorhiza strain, access the PDA of 400mL
In fluid nutrient medium, 28 DEG C, 200rpm is cultivated 10 days, obtains bacterium solution；
(2) peat soil is mixed with PDA liquid medium according to mass ratio 2:1, sterilizes, obtains peat earth mixtures；
(3) the peat earth mixtures that step (2) obtain is mixed with the bacterium solution 2:1 in mass ratio of step (1), obtains bacterial manure.
The present invention has the advantages that Seeds of Dendrobium Candidum is sprouted at present and the conventional method of nursery is by tissue culture come complete
At, and one transfer room that can be run of establishment and tissue culture room are to need many cost inputs to carry out purchase of equipment, and need to spend
A series of working hours such as the sub-bottle cultivation after upper a large amount of working hours complete the production of culture medium, seed is sprouted.Method of the invention is
A kind of sprouting and nursery simple and quick, that seed can be completed in normal environment, eliminate with the equipment of tissue culture method and
Working hour greatly reduces the production cost of seedlings of Dendrobium officinale.
Detailed description of the invention
Fig. 1 is that after planting the 5th day seed expands a turn green sprouting.
Fig. 2 is dendrobium candidum seedling.
Specific embodiment
Embodiment 1
1, the preparation of Epulorhiza solid medium
Epulorhiza is accessed in PDA culture medium, the preparation of the PDA culture medium are as follows: by peeling potatoes, weigh 200g be cut into it is small
Block, be put into it is well-done in 1000ml water smash to pieces, with four layers of filtered through gauze, 20g glucose and 10g agar powder is added in filtrate, is settled to
1000ml, packing sterilizing, is poured into sterile culture dish, every ware 20ml after cooling.
2, endogenetic fungus bacterial manure is prepared
(1) use transfer needle from picking 0.7cm on Epulorhiza solid medium2Epulorhiza strain, access the PDA of 400mL
In fluid nutrient medium, 28 DEG C, 200rpm is cultivated 10 days, obtains bacterium solution；
(2) peat soil is mixed with PDA liquid medium according to mass ratio 2:1, sterilizes, obtains peat earth mixtures；
(3) the peat earth mixtures that step (2) obtain is mixed with the bacterium solution 2:1 in mass ratio of step (1), obtains bacterial manure.
3, bacterium solution liquid is prepared
With transfer needle from picking 0.7cm on Epulorhiza solid medium2Epulorhiza strain, access the PDA liquid of 400mL
In culture medium, 28 DEG C, after 200rpm is cultivated 10 days, thallus is removed with 0.22 μm of membrane filtration to get bacterium solution liquid is arrived.
4, dendrobium candidum nursery
(1) seed is picked: after mature dendrobium candidum fruit pod picking, being gently rinsed with water fruit pod epidermis, is placed on clean white
It is dried on paper in shady place, until fruit pod cracks, collection seed is loaded in seed dispenser；
(2) prepared by matrix: seedling medium peat soil is packed at the 2/3 of seedlings nursing plate, is reloaded into 1/3 endogenetic fungus bacterial manure, it is whole
Flat, watering is wet to matrix；
(3) seed is sowed: the seed of 10 coccolith dry measure used in former times fruit pods is packed into seeding device, gently shaking seeding device keeps dendrobium nobile seed uniform
It sows in stromal surface, one layer of preservative film is covered after having sowed with moistening effect and beats several duck eyes on preservative film；
(4) post-broadcast management: after planting seed plate places the shady place under room temperature, and after planting the 5th day seed, which expands, turns green sprouting,
The water content for keeping surface layer seedling medium is 30-35%, and every 30 days spraying primary filtered bacterium solution liquid, obtains nascent after germination
Dendrobium nobile seedling；
(5) hardening: dendrobium nobile seedling to be come into being is long to 3 true leaves, carries out hardening to nascent dendrobium nobile seedling, continuous hardening 15 days obtains iron
Skin dendrobium nobile seedling, it is 95% that germination percentage, which is calculated, survival rate 90%.
Embodiment 2
1, the preparation of Epulorhiza solid medium
Epulorhiza is accessed in PDA culture medium, the preparation of the PDA culture medium are as follows: by peeling potatoes, weigh 200g be cut into it is small
Block, be put into it is well-done in 1000ml water smash to pieces, with four layers of filtered through gauze, 20g glucose and 10g agar powder is added in filtrate, is settled to
1000ml, packing sterilizing, is poured into sterile culture dish, every ware 20ml after cooling.
2, endogenetic fungus bacterial manure is prepared
(1) use transfer needle from picking 0.7cm on Epulorhiza solid medium2Epulorhiza strain, access the PDA of 400mL
In fluid nutrient medium, 28 DEG C, 200rpm is cultivated 10 days, obtains bacterium solution；
(2) peat soil is mixed with PDA liquid medium according to mass ratio 2:1, sterilizes, obtains peat earth mixtures；
(3) the peat earth mixtures that step (2) obtain is mixed with the bacterium solution 2:1 in mass ratio of step (1), obtains bacterial manure.
3, bacterium solution liquid is prepared
With transfer needle from picking 0.7cm on Epulorhiza solid medium2Epulorhiza strain, access the PDA liquid of 400mL
In culture medium, 28 DEG C, after 200rpm is cultivated 10 days, thallus is removed with 0.22 μm of membrane filtration to get bacterium solution liquid is arrived.
4, dendrobium candidum nursery
(1) seed is picked: after mature dendrobium candidum fruit pod picking, being gently rinsed with water fruit pod epidermis, is placed on clean white
It is dried on paper in shady place, until fruit pod cracks, collection seed is loaded in seed dispenser；
(2) prepared by matrix: seedling medium peat soil is packed at the 1/2 of seedlings nursing plate, is reloaded into 1/2 endogenetic fungus bacterial manure, it is whole
Flat, watering is wet to matrix；
(3) seed is sowed: the seed of 10 coccolith dry measure used in former times fruit pods is packed into seeding device, gently shaking seeding device keeps dendrobium nobile seed uniform
It sows in stromal surface, one layer of preservative film is covered after having sowed with moistening effect and beats several duck eyes on preservative film；
(4) post-broadcast management: after planting seed plate places the shady place under room temperature, and after planting the 5th day seed, which expands, turns green sprouting,
The water content for keeping surface layer seedling medium is 30-35%, and every 30 days spraying primary filtered bacterium solution liquid, obtains nascent after germination
Dendrobium nobile seedling；
(5) hardening: dendrobium nobile seedling to be come into being is long to 3 true leaves, carries out hardening to nascent dendrobium nobile seedling, continuous hardening 15 days obtains iron
Skin dendrobium nobile seedling, it is 87% that germination percentage, which is calculated, survival rate 90%.
Embodiment 3
1, the preparation of Epulorhiza solid medium
Epulorhiza is accessed in PDA culture medium, the preparation of the PDA culture medium are as follows: by peeling potatoes, weigh 200g be cut into it is small
Block, be put into it is well-done in 1000ml water smash to pieces, with four layers of filtered through gauze, 20g glucose and 10g agar powder is added in filtrate, is settled to
1000ml, packing sterilizing, is poured into sterile culture dish, every ware 20ml after cooling.
2, endogenetic fungus bacterial manure is prepared
(1) use transfer needle from picking 0.7cm on Epulorhiza solid medium2Epulorhiza strain, access the PDA of 400mL
In fluid nutrient medium, 28 DEG C, 200rpm is cultivated 10 days, obtains bacterium solution；
(2) peat soil is mixed with PDA liquid medium according to mass ratio 2:1, sterilizes, obtains peat earth mixtures；
(3) the peat earth mixtures that step (2) obtain is mixed with the bacterium solution 2:1 in mass ratio of step (1), obtains bacterial manure.
3, bacterium solution liquid is prepared
With transfer needle from picking 0.7cm on Epulorhiza solid medium2Epulorhiza strain, access the PDA liquid of 400mL
In culture medium, 28 DEG C, after 200rpm is cultivated 10 days, thallus is removed with 0.22 μm of membrane filtration to get bacterium solution liquid is arrived.
4, dendrobium candidum nursery
(1) seed is picked: after mature dendrobium candidum fruit pod picking, being gently rinsed with water fruit pod epidermis, is placed on clean white
It is dried on paper in shady place, until fruit pod cracks, collection seed is loaded in seed dispenser；
(2) prepared by matrix: seedling medium peat soil is packed at the 3/4 of seedlings nursing plate, is reloaded into 1/4 endogenetic fungus bacterial manure, it is whole
Flat, watering is wet to matrix；
(3) seed is sowed: the seed of 10 coccolith dry measure used in former times fruit pods is packed into seeding device, gently shaking seeding device keeps dendrobium nobile seed uniform
It sows in stromal surface, one layer of preservative film is covered after having sowed with moistening effect and beats several duck eyes on preservative film；
(4) post-broadcast management: after planting seed plate places the shady place under room temperature, and after planting the 5th day seed, which expands, turns green sprouting,
The water content for keeping surface layer seedling medium is 30-35%, and every 20 days spraying primary filtered bacterium solution liquid, obtains nascent after germination
Dendrobium nobile seedling；
(5) hardening: dendrobium nobile seedling to be come into being is long to 3 true leaves, carries out hardening to nascent dendrobium nobile seedling, continuous hardening 15 days obtains iron
Skin dendrobium nobile seedling, it is 91% that germination percentage, which is calculated, survival rate 89%.
The foregoing is merely presently preferred embodiments of the present invention, all equivalent changes done according to scope of the present invention patent with
Modification, is all covered by the present invention.
Claims (4)
1. a kind of method for culturing seedlings of dendrobium candidum endogenetic fungus, which is characterized in that cultivate dendrobium candidum using endogenetic fungus bacterial manure
Seed makes fungi and Seed Development symbiosis, without the sprouting and nursery of seed can be realized by tissue culture.
2. a kind of method for culturing seedlings of dendrobium candidum endogenetic fungus according to claim 1, which is characterized in that specifically include as
Lower step:
(1) seed is picked: after mature dendrobium candidum fruit pod picking, being gently rinsed with water fruit pod epidermis, is placed on clean white
It is dried on paper in shady place, until fruit pod cracks, collection seed is loaded in seed dispenser；
(2) prepared by matrix: seedling medium peat soil is packed at the 1/2-3/4 of seedlings nursing plate, then filled up with endogenetic fungus bacterial manure, it is whole
Flat, watering is wet to matrix；
(3) seed is sowed: gently shaking seeding device sows dendrobium nobile seed uniformly in stromal surface, and one layer of guarantor is covered after having sowed
Fresh film has moistening effect and beats several duck eyes on preservative film；
(4) post-broadcast management: after planting seed plate places the shady place under room temperature, and the water content of holding surface layer seedling medium is
30-35%, every 20-30 days spraying primary filtered bacterium solution liquid, obtains nascent dendrobium nobile seedling after germination；
(5) hardening: dendrobium nobile seedling to be come into being is long to 3 true leaves, carries out hardening to nascent dendrobium nobile seedling, continuous hardening 15 days obtains iron
Skin dendrobium nobile seedling.
3. a kind of method for culturing seedlings of dendrobium candidum endogenetic fungus according to claim 2, which is characterized in that the step
(4) bacterium solution liquid the preparation method is as follows: with transfer needle from picking 0.7cm on Epulorhiza solid medium in2Tumor mycorhiza
Bacterium strain, is accessed in the PDA liquid medium of 400mL, 28 DEG C, after 200rpm is cultivated 10 days, is removed with 0.22 μm of membrane filtration
Thallus to get arrive bacterium solution liquid.
4. a kind of method for culturing seedlings of dendrobium candidum endogenetic fungus according to claim 2, it is characterised in that the Nei Shengzhen
Bacterium bacterial manure the preparation method is as follows:
(1) use transfer needle from picking 0.7cm on Epulorhiza solid medium2Epulorhiza strain, access the PDA liquid of 400mL
In body culture medium, 28 DEG C, 200rpm is cultivated 10 days, obtains bacterium solution；
(2) peat soil is mixed with PDA liquid medium according to mass ratio 2:1, sterilizes, obtains peat earth mixtures；
(3) the peat earth mixtures that step (2) obtain is mixed with the bacterium solution 2:1 in mass ratio of step (1), obtains bacterial manure.
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| CN201811042090.5A CN109122206A (en) | 2018-09-07 | 2018-09-07 | A kind of method for culturing seedlings of dendrobium candidum endogenetic fungus |
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| CN201811042090.5A CN109122206A (en) | 2018-09-07 | 2018-09-07 | A kind of method for culturing seedlings of dendrobium candidum endogenetic fungus |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109804913A (en) * | 2019-01-17 | 2019-05-28 | 山西大学 | A kind of construction method of yellow bluestem-endogenetic fungus homobium |
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| CN107432240A (en) * | 2017-08-01 | 2017-12-05 | 霍山县大化坪秀峰茶业精制厂 | A kind of method for culturing seedlings of Dendrobidium huoshanness |
| CN107827616A (en) * | 2017-12-15 | 2018-03-23 | 福建省农业科学院亚热带农业研究所 | A kind of stem of noble dendrobium artificial growth organic fungi-manure and preparation method thereof |
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2018
- 2018-09-07 CN CN201811042090.5A patent/CN109122206A/en active Pending
Patent Citations (3)
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| CN107396725A (en) * | 2017-07-18 | 2017-11-28 | 杭州千岛湖峰川生物科技有限公司 | A kind of dendrobium candidum implantation methods |
| CN107432240A (en) * | 2017-08-01 | 2017-12-05 | 霍山县大化坪秀峰茶业精制厂 | A kind of method for culturing seedlings of Dendrobidium huoshanness |
| CN107827616A (en) * | 2017-12-15 | 2018-03-23 | 福建省农业科学院亚热带农业研究所 | A kind of stem of noble dendrobium artificial growth organic fungi-manure and preparation method thereof |
Non-Patent Citations (1)
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109804913A (en) * | 2019-01-17 | 2019-05-28 | 山西大学 | A kind of construction method of yellow bluestem-endogenetic fungus homobium |
| CN109804913B (en) * | 2019-01-17 | 2021-09-28 | 山西大学 | Method for constructing aegilops tauschii-endophytic fungi symbiont |
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