CN106386482B - Dendrobium candidum high quality seedling nature optical culture once-seedling forming quick propagating method - Google Patents
Dendrobium candidum high quality seedling nature optical culture once-seedling forming quick propagating method Download PDFInfo
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- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 3
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- 239000011159 matrix material Substances 0.000 claims description 3
- 239000002609 medium Substances 0.000 claims description 3
- 229960002523 mercuric chloride Drugs 0.000 claims description 3
- LWJROJCJINYWOX-UHFFFAOYSA-L mercury dichloride Chemical compound Cl[Hg]Cl LWJROJCJINYWOX-UHFFFAOYSA-L 0.000 claims description 3
- YGGXZTQSGNFKPJ-UHFFFAOYSA-N methyl 2-naphthalen-1-ylacetate Chemical compound C1=CC=C2C(CC(=O)OC)=CC=CC2=C1 YGGXZTQSGNFKPJ-UHFFFAOYSA-N 0.000 claims description 3
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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
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- 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 dendrobium candidum high quality seedling nature optical culture once-seedling forming quick propagating methods, are mainly made of 6 steps.It is followed successively by and prepares dendrobium candidum material, build full natural light culturing room, make seed suspension liquid, aseptic seeding, once-seedling forming culture and the transplanting of test tube seedling.Wherein once-seedling forming culture is further divided into protocorm Fiber differentiation, seedling culture and stock breeding three phases.Further include the once-seedling forming culture medium used in above-mentioned rapid propagation method.The present invention can be with once-seedling forming, it is artificial to be not required to switching, in addition use full natural light culturing room, the position of culture is moved according to demand difference of the different stages of growth to light, the seedling stalwartness produced easily survives, special hardening treatment is needed not move through before transplanting, can directly bottle outlet transplant, significantly reduce production cost.The large-scale production of seedlings of Dendrobium officinale once-seedling forming can be realized using the seed of high germination rate, culture medium full of nutrition and full natural light culturing room.
Description
Technical field
The invention belongs to the plant regeneration technologies by tissue culture technique, and in particular to a kind of dendrobium candidum high quality seedling
Natural optical culture once-seedling forming quick propagating method and used once-seedling forming culture medium.
Background technology
Dendrobium candidum is orchid family Dendrobium epiphyte, is exactly generally acknowledged rare Chinese herbal medicine since ancient times.Wild iron sheet stone
Dry measure used in former times environmental condition needed for growth and development is extremely harsh, slow-growing, and breeding potential is extremely low, and artificial predatory excavation has been directed at open country
Pig iron skin Dendrobium Plants are bordering on exhaustion.Current propagation method is mainly divided to two classes, and one kind is tissue rapid propagation.It should before tissue-cultured seedling cultivation
The hardening of 14~21 days is first carried out, concrete measure is that tissue-cultured seedling is moved to hardening room, is made in its environment in open variation gradually
Reform of nature environment, can bottle outlet plantation when tissue-cultured seedling leaf color is dark green.First bottle cap is opened before bottle outlet, allows bottle seedling outdoor empty
It is placed 2~3 days in gas, allows its reform of nature humiture.Then it carries out washing seedling, gently takes the seedling with culture medium when washing seedling
Go out, be placed in basin and clean, pollute seedling and bare-root seeding or few offspring should be separated.Tissue-cultured seedling is first cleaned with tap water, mainly
Agar should be washed off, in order to avoid agar is mouldy to cause rotten, is then rinsed with clear water again.When washing seedling preferably based on the size of seedling and
Quality, which synchronizes, to be classified, and to cultivate, to manage, is improved the survival rate of tissue-cultured seedling and is made growth neat.Another kind of is plant division
Cuttage, selected when cardinal principle is plant division life in 1 year or life in 2 years, color and luster it is light green, it is healthy and strong, sprout more, well developed root system and no disease and pests harm
Plant make kind of a strain, cut off deadwood, disconnected branch, old branch and long fibrous root, strain clump cut, be divided into little Cong, every clump with leaf
Stem strain 5~7, you can plantation.
Although the technology of tissue culture breeding seedlings of Dendrobium officinale has succeeded, seedlings of Dendrobium officinale generally existing culture
The problems such as period is long, pollution rate is high, complicated for operation, production cost is higher, so as to restrict the quick hair of dendrobium candidum industry
Exhibition.As Publication No. CN101564008A, it is entitled " a kind of dendrobium candidum axenic seedlings without hormone culture and quick breeding side
The domestic patent of invention of method " disclose a kind of dendrobium candidum axenic seedlings without hormone culture and rapid propagation method, content includes
The formation and preservation of the axenic germination of Seeds of Dendrobium Candidum, protocorm and seedling, the taking root of aseptic seedling are proliferated and strong seedling culture,
The processes such as the white silk seedling of aseptic seedling and transplanting.But the technology have the drawback that the Induction Process of protocorm and seedling be related to feux rouges,
The use of green light, unglazed, yellow light and blue light, therefore process is extremely complex, and the process induced is difficult control, is unfavorable in big
Technical scale production application
Invention content
The technical problem to be solved by the present invention is to the cultivation cycle length of seedlings of Dendrobium officinale generally existing, pollution rate height, behaviour
The problems such as making the problem of complicated, production cost is higher, seed of the present invention by using high germination rate, culture full of nutrition
Base, full natural light culturing room realize the large-scale production of seedlings of Dendrobium officinale once-seedling forming, have inexpensive, efficient feature,
It is the High biotechnology of the large-scale production of seedling, the sustainable development of dendrobium candidum industry can be promoted.
To solve the above problems, technical solution proposed by the present invention is a kind of dendrobium candidum high quality seedling nature optical culture one
Secondary seedling forming quick propagating method, comprises the steps of：
(1) prepare dendrobium candidum material；
(2) full natural light culturing room is built
Using translucent material, and shading system, cooling system and culturing rack are equipped with, are ensured in culturing room by shading system
The intensity of illumination of top layer culturing rack is 3000~5000Lux, and intensity of illumination is as the number of plies of culturing rack moves down and reduces, bottom training
The intensity of illumination for supporting frame is 100~1000Lux, and cooling system can control the temperature of glass culturing room at 20~30 DEG C, culture
Frame length can be depending on the size of culturing room；
(3) seed suspension liquid is made
In dendrobium candidum field gene bank, the strong healthy strain of robust growth, germination rate height, resistance, disease and insect resistance is chosen,
Artificial pollination and bagging are carried out in season of flowers, the capsule for choosing a certain range fruit age is explant, carries out disinfection, cleans and removes
Water then cuts capsule with aseptic operation knife, the seed in capsule is all eluted in a bottle equipped with sterile water, each capsule
The seed correspondence of fruit is eluted to a bottle, and seed suspension liquid is made, the inspection of seed density is carried out to each capsule seed suspension liquid
Survey, be further diluted to sterile water according to every bottle of suspension seed concentration seed it is 10-15 a concentration of/inoculation of milliliter
Seed suspension liquid；
(4) aseptic seeding
2 milliliters of the seed suspension liquid of inoculation is drawn to equipped with 150 milliliters of once-seedling forming with liquid-transfering gun under aseptic condition
In the orchid culture bottle of solid medium, the seed amount of every bottle of dendrobium candidum 20-30；
(5) once-seedling forming culture
Protocorm Fiber differentiation：Seed suspension liquid is uniformly distributed in media surface, this culture bottle is placed complete natural
On culturing rack in optical culture room, its intensity of illumination is kept in 100~1000Lux, keeps culture room temperature in (26 ± 2) DEG C,
It is cultivated 60~90 days under the conditions of this, seed is sprouted to form protocorm, and protocorm sends out roots and blade；
Seedling culture：Seedling is moved to the suitable layers of culturing rack, strengthens its natral light cahure, promotes seedling growth,
This period incubation time is 90 days, and cultivation temperature is (26 ± 2) DEG C, and seedling grows up to seedlings, height of seedling 3~5cm, 3~5~leaf
Piece, 3~5 roots, 1~3 centimetre of root long；Seedlings base portion sprouts 1-2 new talent, forms tufted seedling；
Stock breeding：Seedlings are moved to the top layer of culturing rack, is cultivated under high natural light light intensity, can both promoted
The growth of seedlings, and can achieve the effect that hardening, this period cultivates 60 days, and cultivation temperature is (26 ± 2) DEG C, at this time iron sheet stone
Every clump of 2-5 seedling of dry measure used in former times seedling, 5~8cm of height of seedling have 4-6 sections, and 4 reach with blade, 3 or more roots, thick more than the 2.0mm of stem
The standard of seedling；
(6) transplanting of test tube seedling
During transplanting, rooted seedling is taken out from culture bottle, it is molten with millesimal potassium permanganate after the culture medium for cleaning attachment
Liquid impregnates 5min, then dries rooted seedling to root and bleaches, and planting matrix uses sterile-processed bark, pays attention to keeping suitable
Suitable humidity and temperature are placed at shady and cool ventilation and cultivate, and the plant after surviving for 30 days or so can generate new root system, may move at this time
Greenhouse carries out normal water, fertilizer, pencil reason.
Further, in step (2), it is assumed that culturing rack is four-layer structure, then second and third layer of culturing rack illumination of top layer down
Intensity is 1000~3000Lux.
Preferably, a certain range fruit age described in step (3) is 120 days.
Further, detailed process is sterilized, cleaned and removed water described in step (3) is：It is impregnated 30 seconds in 70% alcohol, then
It is sterilized 15~20 minutes with 0.1% mercuric chloride solution, aseptic water washing 4~5 times, with sterile under aseptic condition on superclean bench
Filter paper blots the moisture on capsule surface.
Further, the detection of seed density described in above-mentioned steps (3) refers to plant each capsule seed suspension liquid
The detection of sub- density, detailed process are to draw 20 microlitres with sterile liquid-transfering gun and under biomicroscope calculate it in the case that shaking up
Seed amount, every bottle of seed suspension liquid, which repeats to take 10 times, calculates its average every milliliter of seed number.
In addition, the present invention also provides a kind of quick sides of breeding of above-mentioned dendrobium candidum high quality seedling nature optical culture once-seedling forming
The once-seedling forming culture medium used in method, it includes following component：Every liter, containing No. 1 1~2g of treasured is spent, is spent No. 2 1~2g of treasured, albumen
0.5~2g of peptone, 40~80g of mashed potatoes, 50~100g of banana puree, 10~20g of apple butter, 500~1000mg of activated carbon, inositol 80
~120mg, 1.5~2.5mg of glycine, thiamine hydrochloride (VB1) 0.05~0.2mg, puridoxine hydrochloride (VB6) 0.4~
0.05~0.1 milligram, methyl α-naphthyl acetate (NAA) 0.2~2mg, 15~30g of sucrose of 0.8mg, niacin 0.4~0.8mg, 6- benzyl purine,
Agar 6~7g, pH 5.4-5.6.
Compared with prior art, advantage of the invention is that：
1, can switching be not required to manually with once-seedling forming, in addition full natural light culturing room is used, according to different stages of growth pair
The demand of light is different and moves the position of culture, and the seedling produced is healthy and strong easily to be survived, and is needed not move through before transplanting special
Hardening treatment can directly bottle outlet transplant, significantly reduce production cost.
2, carry out seedlings of Dendrobium officinale using the seed of high germination rate, culture medium full of nutrition, full natural light culturing room
Once-seedling forming mass produces, and without disease pest robust growth, seedling has good quality, resistance, grows rapidly, convenient for life test tube seedling
Production management and the advantages such as production cost are reduced, can realize timing, determine that matter, quantitatively offer specification is standardized, neat and consistent high-quality
Dendrobium candidum test tube seedling is to meet the market demand.
Description of the drawings
Fig. 1 is the flow chart of the present invention.
Specific embodiment
The specific embodiment of the present invention is described in further detail in conjunction with attached drawing.
The flow chart of the present invention 6 steps as shown in Figure 1, be mainly made of.It is followed successively by and prepares dendrobium candidum material, build
Full natural light culturing room, makes seed suspension liquid, aseptic seeding, once-seedling forming culture and the transplanting of test tube seedling.Wherein once into
Seedling culture is further divided into protocorm Fiber differentiation, seedling culture and stock breeding three phases.
To cause those of ordinary skill in the art that the present invention may be better understood, a specific implementation is now provided
Example is as follows：
1. prepare dendrobium candidum (Dendrobium officinale Kimura et Migo) material.
2. full natural light culturing room builds：The culturing room of light transmission is built up using tempered glass, is equipped with shading system, cooling is set
Standby, culturing rack.Shading system includes inside and outside sunshade net, and the illumination of culture building overlay culturing rack is made by sunshade net inside and outside regulation and control
Intensity is 3000~5000Lux, second and third layer of culturing rack intensity of illumination of top layer down is 1000~3000Lux, and bottom pushes up
The lower 4th layer of 100~1000Lux of intensity of illumination of layer.Cooling system uses air conditioner for both cooling and heating, can be by the temperature of glass culturing room
Control is at 20~30 DEG C.Culturing rack width be 60 centimetres, a height of 200 centimetres, from the ground 10 centimetres be the 4th layer, layer away from 60 centimetres,
Totally four layers, culturing rack length can be depending on the size of culturing room.
3. the making of seed suspension liquid：In dendrobium candidum field gene bank, choose robust growth, germination rate height, resistance, resist
The strong healthy strain of pest and disease damage carries out artificial pollination and bagging in season of flowers, and it is explant to choose the capsule that fruit age is 120 days
Body impregnates 30 seconds in 70% alcohol, then is sterilized 15~20 minutes with 0.1% mercuric chloride solution, aseptic water washing 4~5 times, Yu Chao
Blot the moisture on capsule surface on net workbench under aseptic condition with aseptic filter paper.Then capsule is cut with aseptic operation knife, it will
Seed in capsule is all eluted in a triangular flask equipped with 200 milliliters of sterile waters, and the seed correspondence of each capsule is eluted to
The one bottled triangular flask for having 200 milliliters of sterile waters, is made seed suspension liquid.Seed density is carried out to each capsule seed suspension liquid
Detection, in the case that shake up with sterile liquid-transfering gun draw 20 microlitres its seed amount, every bottle of kind are calculated under biomicroscope
Sub- suspension, which repeats to take 10 times, calculates its average every milliliter of seed number.It is further according to every bottle of suspension seed concentration sterile water
Be diluted to seed it is 10-15 a concentration of/the seed suspension liquid of the inoculation of milliliter.
4. aseptic seeding：2 milliliters of the seed suspension liquid of inoculation is drawn to equipped with 150 milliliters with liquid-transfering gun under aseptic condition
Once-seedling forming solid medium orchid culture bottle in, the seed amount of every bottle of dendrobium candidum 20-30.Once-seedling forming culture
Base：Every liter, containing No. 1 1.5g of treasured is spent, is spent treasured No. 2 1.5g, peptone 2g, mashed potatoes 40g, banana puree 80g, apple butter 10g, activity
500~1000mg of carbon, inositol 100mg, glycine 2.0mg, thiamine hydrochloride (VB1) 0.2mg, puridoxine hydrochloride (VB6)
0.05 milligram, methyl α-naphthyl acetate (NAA) 0.5mg, sucrose 20g, agar 6g, pH 5.4- of 0.5mg, niacin 0.5mg, 6- benzyl purine
5.6。
5. once-seedling forming culture：
Protocorm Fiber differentiation：Seed suspension liquid is uniformly distributed in media surface, this culture bottle is placed complete natural
On the 4th floor culturing rack in optical culture room, keep its intensity of illumination in 100~1000Lux, keep culture room temperature (25 ±
2)℃.It is cultivated 60~90 days under the conditions of this, seed is sprouted to form protocorm, and protocorm sends out roots and blade, and cultivation temperature is (26
±2)℃。
Seedling culture：Seedling is moved to second or third layer of culturing rack, strengthens its natral light cahure, promotes seedling
Growth, this period incubation time are 90 days, and seedling grows up to seedlings, height of seedling 3~5cm, 3~5~blade, 3~5 roots, root
It is 1~3 centimetre long；Seedlings base portion sprouts 1-2 new talent, forms tufted seedling, and cultivation temperature is (26 ± 2) DEG C.
Stock breeding：Seedlings are moved to the first layer of culturing rack, is cultivated under high natural light light intensity, can both promoted
Into the growth of seedlings, and it can achieve the effect that hardening.This period cultivates 60 days, every clump of 2-5 seedling of dendrobium candidum seedling at this time, seedling
High 5~8cm, there is a 4-6 sections, 4 with blade, 3 or more roots, thick more than the 2.0mm of stem reaches the standard of emergence, cultivation temperature
For (26 ± 2) DEG C.
6. the transplanting of test tube seedling：During transplanting, rooted seedling is taken out from culture bottle, after the culture medium for cleaning attachment, with thousand points
One of liquor potassic permanganate impregnate 5min, then rooted seedling is dried to root and is bleached, planting matrix use is sterile-processed
Bark, pay attention to keeping suitable for humidity and temperature, be placed at shady and cool ventilation and cultivate, survival rate is up to more than 98%.30 days or so
Plant after surviving can generate new root system, may move into greenhouse at this time and carry out normal water, fertilizer, pencil reason.
The foregoing is merely the specific embodiments of the present invention, are not intended to limit the invention, all essences in the present invention
With within principle, any modification, equivalent replacement, improvement and so on should all be included in the protection scope of the present invention god.
Claims (4)
1. dendrobium candidum high quality seedling nature optical culture once-seedling forming quick propagating method, it is characterised in that comprise the steps of：
（1）Prepare dendrobium candidum material；
（2）Build full natural light culturing room
Using translucent material, and shading system, cooling system and culturing rack are equipped with, ensure to cultivate building overlay by shading system
The intensity of illumination of culturing rack is 3000~5000 Lux, and intensity of illumination is as the number of plies of culturing rack moves down and reduces, bottom culture
The intensity of illumination of frame is 100~1000 Lux, second and third layer of culturing rack intensity of illumination of top layer down is 1000~3000
Lux, cooling system control the temperature of glass culturing room at 20~30 °C, and culturing rack length is depending on the size of culturing room；
（3）Make seed suspension liquid
In dendrobium candidum field gene bank, the strong healthy strain of robust growth, germination rate height, resistance, disease and insect resistance, Yu Kai are chosen
Adolescence carries out artificial pollination and bagging, and the capsule for choosing a certain range fruit age is explant, carries out disinfection, cleans and removes water,
Then capsule is cut with aseptic operation knife, the seed in capsule is all eluted in a bottle equipped with sterile water, each capsule
Seed correspondence be eluted to a bottle, seed suspension liquid is made, to each capsule seed suspension liquid carry out seed density detection,
Further be diluted to sterile water according to every bottle of suspension seed concentration seed it is 10-15 a concentration of/kind of the inoculation of milliliter
Sub- suspension；
（4）Aseptic seeding
2 milliliters of the seed suspension liquid of inoculation is drawn to equipped with 150 milliliters of once-seedling forming solid with liquid-transfering gun under aseptic condition
In the orchid culture bottle of culture medium, the seed amount of every bottle of dendrobium candidum 20-30, the once-seedling forming solid medium into
Divide as follows：No. 1 1~2g/L of treasured is spent, spends treasured No. 2 1~2g/ L, peptone 0.5~2g/ L, 40~80 g/ L of mashed potatoes, it is fragrant
Any of several broadleaf plants mud 50~100g/ L, apple butter 10~20g/ L, activated carbon 500 ~ 1000mg/ L, inositol 80 ~ 120mg/ L, glycine
1.5 ~ 2.5mg/ L, thiamine hydrochloride 0.05 ~ 0.2mg/ L, puridoxine hydrochloride 0.4 ~ 0.8mg/ L, niacin 0.4 ~ 0.8mg/ L,
6- benzyl purine 0.05 ~ 0.1mg/ L, methyl α-naphthyl acetate 0.2 ~ 2mg/ L, sucrose 15~30g/ L, 6~7g/ of agar L, pH 5.4-
5.6；
（5）Once-seedling forming culture
Protocorm Fiber differentiation：Seed suspension liquid is uniformly distributed in media surface, this culture bottle is placed into full natural light and is trained
It supports on the culturing rack in room, keeps its intensity of illumination in 100~1000 Lux, keep culture room temperature in (26 ± 2) DEG C, this
It is cultivated 60~90 days under part, seed is sprouted to form protocorm, and protocorm sends out roots and blade；
Seedling culture：Seedling is moved to the suitable layers of culturing rack, strengthens its natral light cahure, promotes seedling growth, at this time
Phase incubation time is 90 days, and cultivation temperature is (26 ± 2) DEG C, and seedling grows up to seedlings, 3~5cm of height of seedling, 3~5 blades, 3~5
Root, 1~3 centimetre of root long；Seedlings base portion sprouts 1-2 new talent, forms tufted seedling；
Stock breeding：Seedlings are moved to the top layer of culturing rack, is cultivated under high natural light light intensity, can both promote seedlings
Growth, and can achieve the effect that hardening, this period cultivates 60 days, and cultivation temperature is (26 ± 2) DEG C, at this time dendrobium candidum seedling
Every clump of 2-5 seedling, 5~8cm of height of seedling have 4-6 sections, and 4 reach emergence with blade, 3 or more roots, thick more than the 2.0mm of stem
Standard；
（6）The transplanting of test tube seedling
During transplanting, rooted seedling is taken out from culture bottle, after the culture medium for cleaning attachment, is soaked with millesimal liquor potassic permanganate
5 min are steeped, then rooted seedling is dried to root and is bleached, planting matrix uses sterile-processed bark, pays attention to keeping suitable
Humidity and temperature are placed at shady and cool ventilation and cultivate, and the plant after surviving for 30 days or so can generate new root system, move at this time big
Canopy carries out normal water, fertilizer, pencil reason.
2. dendrobium candidum high quality seedling nature optical culture once-seedling forming quick propagating method according to claim 1, special
Sign is step（3）Described in a certain range fruit age be preferably 120 days.
3. dendrobium candidum high quality seedling nature optical culture once-seedling forming quick propagating method according to claim 1, special
Sign is step（3）Described in disinfection, cleaning and water removal be specially to be impregnated 30 seconds in 70% alcohol, then with 0.1% mercuric chloride solution
Disinfection 15 ~ 20 minutes, aseptic water washing 4 ~ 5 times blots capsule surface under aseptic condition on superclean bench with aseptic filter paper
Moisture.
4. dendrobium candidum high quality seedling nature optical culture once-seedling forming quick propagating method according to claim 1, special
Sign is step（3）Described in seed density detection refer to each capsule seed suspension liquid carry out seed density detection,
Detailed process is to draw 20 microlitres with sterile liquid-transfering gun and calculate its seed amount under biomicroscope in the case that shaking up, every bottle
Seed suspension liquid, which repeats to take 10 times, calculates its average every milliliter of seed number.
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| CN107182735B (en) * | 2017-04-27 | 2019-10-25 | 江西九草铁皮石斛科技协同创新有限公司 | A kind of method for culturing seedlings and cultural method of live streaming formula dendrobium candidum |
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| CN107581067A (en) * | 2017-09-29 | 2018-01-16 | 南京仙草堂生物科技有限公司 | A kind of little Hua all ages orchid species seedling aseptic seeding propagation methods |
| CN109089886A (en) * | 2018-09-18 | 2018-12-28 | 广东兴源农林有限公司 | A kind of dendrobium candidum seedling culture medium |
| CN109924130B (en) * | 2019-04-22 | 2021-03-23 | 南京农业大学 | Method for rapidly propagating dendrobium officinale seedlings |
| CN111837961A (en) * | 2020-08-10 | 2020-10-30 | 南京农业大学 | Dendrobium officinale seedling culture medium and seedling culture method |
| CN111919749B (en) * | 2020-08-20 | 2022-03-25 | 南京农业大学 | Culture medium for rapid propagation of dendrobium nobile seedlings and rapid propagation method thereof |
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