CN108617511A - The seed group fast tissue culture reproducing method of dendrobium candidum - Google Patents
The seed group fast tissue culture reproducing method of dendrobium candidum Download PDFInfo
- Publication number
- CN108617511A CN108617511A CN201810909426.7A CN201810909426A CN108617511A CN 108617511 A CN108617511 A CN 108617511A CN 201810909426 A CN201810909426 A CN 201810909426A CN 108617511 A CN108617511 A CN 108617511A
- Authority
- CN
- China
- Prior art keywords
- seedling
- culture
- seed
- dendrobium candidum
- rooting
- 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
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
Abstract
The present invention proposes a kind of seed group fast tissue culture reproducing method of dendrobium candidum, includes the following steps：150 days fruit ages tied on the dendrobium candidum of robust growth or more uncracked ripe capsule is selected, is cleaned, drained, sterilized；It is longitudinally splitted after cutting off each 0.3~0.7cm in its head and the tail both ends, takes out seed, seed is uniformly inoculated on seed sprouting differential medium and cultivated；When seed is sprouted and is differentiated to form the seedling of two panels leaflet, the seedling switching of gained is subjected to Multiplying culture in seedling proliferation culture medium, obtains tufted seedling；It when tufted seedling is grown to 2 3cm high, is split tufted seedling to form single plant seedling from base portion, is transferred on Rooting and hardening-off culture base and is cultivated；When seedling grow to 4.0~5.0cm high and grow length more than or equal to 2cm root at least 2 when, acquisition can bottle outlet plant seedling.Method using the present invention can quickly and easily obtain the candidum tissue culturing seedling of a large amount of low costs.
Description
Technical field
The present invention relates to biotechnologies, and in particular to a kind of seed group fast tissue culture reproducing method of dendrobium candidum.
Background technology
Dendrobium candidum is a kind of traditional rare rare traditional Chinese medicine, because with antitumor, anti-aging, acoustic resistive band fatigue, expanding
The effects that opening blood vessel and enhancing human immunity is referred to as " help celestial grass ", ranks first of " Chinese nine big celestial grass ".With iron sheet
Stem of noble dendrobium health value is continuously developed, and medicinal material market is also gradually increasing the demand of dendrobium candidum, but due to excessively adopting for a long time
It plucks, Chinese wild resource is exhausted, and low with traditional vegetative manner breeding coefficient, cannot be satisfied commerial growing
Seedling needs, and tissue-cultured seedling has become the main source of seedling of current dendrobium candidum plantation at present.Currently to candidum tissue culturing
Fast breeding technique research is more, but most of there are medium components complicated, growing-seedling period compared with long, labor intensive is more, tissue-cultured seedling at
The problems such as this is high seriously constrains the industrialized development of dendrobium candidum.
To sum up, it needs to be improved the prior art.
Invention content
The technical problem to be solved in the present invention is to provide a kind of quick breeding by group culture methods of dendrobium candidum, using the present invention
Method can quickly and easily obtain the candidum tissue culturing seedlings of a large amount of low costs.
In order to solve the above technical problems, the present invention proposes a kind of seed group fast tissue culture reproducing method of dendrobium candidum, successively
Include the following steps：
1) it, draws materials：
150 days fruit ages tied on the dendrobium candidum of robust growth or more uncracked ripe capsule is selected, cleaned, dripped
It is spare after dry；
2), the induction of seed is sprouted and is broken up：
After the capsule obtained by step 1) sterilized (routine disinfection), longitudinal direction after each 0.3~0.7cm in its head and the tail both ends is cut off
It splits, takes out seed, seed is uniformly inoculated on seed sprouting differential medium and is cultivated；
Condition of culture is：
Light culture is carried out first 10 ± 1 days, temperature is 25 ± 1 DEG C；
Carry out illumination in 14 hours, 30~40 μm of ol m of intensity of illumination daily later-2·s-1, temperature is 25 ± 1 DEG C；10 is small
When light culture, temperature be 21 ± 1 DEG C；Above-mentioned illumination and light culture are alternately；
3), the proliferation of seedling：
When seed is sprouted and is differentiated to form seedling (unrooted) of two panels leaflet, the seedling switching of gained is increased in seedling
It grows culture medium and carries out Multiplying culture, obtain tufted seedling；
Condition of culture is：Illumination in 16 hours, 30~40 μm of olm of intensity of illumination-2s-1, temperature is 25 ± 1 DEG C；Dark training in 8 hours
It supports, temperature is 21 ± 1 DEG C；Above-mentioned illumination and light culture are alternately；
4), strong plantlets and rootage：
When the tufted seedling that Multiplying culture obtains is grown to 2-3cm high, tufted seedling is split from base portion to form single plant small
Seedling is transferred on Rooting and hardening-off culture base and carries out Rooting and hardening-off culture；
Condition of culture is：Illumination in 16 hours, 30~40 μm of oLm of intensity of illumination-2s-1, temperature is 25 ± 1 DEG C；Dark training in 8 hours
It supports, temperature is 21 ± 1 DEG C；Above-mentioned illumination and light culture are alternately；
5) it, transplants：
The seedling that under growth root strong seedling culture obtains is grown to 4.0~5.0cm high and grows root of the length more than or equal to 2cm extremely
When 2 few, acquisition can bottle outlet plantation seedling.
The improvement of quick breeding by group culture method as dendrobium candidum of the present invention：
Seed in the step 2) sprouts differential medium：25~35g/L of MS+NAA0.05~0.15mg/L+ sucrose
5~10g/L+0.3 of+agar~0.7g/L activated carbons, pH are 5.5~6.
The specific preparation method that seed sprouts differential medium is as follows：Based on MS minimal mediums, it is separately added into naphthalene
Acetic acid (NAA), sucrose, agar and activated carbon uniformly mix, and it is 5.5 to adjust pH using the HCl of the KOH or 1mol/L of 1mol/L
~6.0；Add 0.05~0.15mgNAA, 25~35g sucrose, 5~10g agar and 0.3~0.7g in MS minimal mediums per 1L
Activated carbon.
Quick breeding by group culture further improvements in methods as dendrobium candidum of the present invention：
Seedling proliferation culture medium in step 3) is：MS+NAA0.15~0.2mg/L+ 25~35g/L+ of sucrose agar 5~
10g/L+ activated carbons 0.3~0.7g/L, pH are 5.5~6.
The specific preparation method of seedling proliferation culture medium is as follows：Based on MS minimal mediums, it is separately added into methyl α-naphthyl acetate
(NAA), sucrose, agar and activated carbon, uniformly mix, using 1mol/L KOH or 1mol/L HCl adjust pH be 5.5~
6.0；In MS minimal mediums per 1L plus 0.15~0.2mgNAA, 25~35g sucrose, 5~10g agar and 0.3~0.7g live
Property charcoal.
Quick breeding by group culture further improvements in methods as dendrobium candidum of the present invention：
Rooting and hardening-off culture base in step 4) be MS+BA0.3~0.7mg/L+NAA0.3~0.7mg/L+ sucrose 25~
35g/L+ agar 5~10g/L+ activated carbons 0.3~0.7g/L, pH are 5.5~6.
The specific preparation method of Rooting and hardening-off culture base is as follows：Based on MS minimal mediums, it is separately added into benzyl amino
Adenine (BA), methyl α-naphthyl acetate (NAA), sucrose, agar and activated carbon uniformly mix, and utilize the KOH or 1mol/L of 1mol/L
It is 5.5~6.0 that HCl, which adjusts pH,；Add 0.3~0.7mgBA, 0.3~0.7mgNAA, 25~35g in MS minimal mediums per 1L
Sucrose, 5~10g agar and 0.3~0.7g activated carbons.
Note：BA is 6-BA (6- benzyls aminoadenine).
Quick breeding by group culture further improvements in methods as dendrobium candidum of the present invention：
First by gained can bottle outlet plantation seedling hardening 5 in the shading greenhouse of quasi- rooting culture is transferred to culture vessel
~7 days；
Then bottle cap is opened, is watered in Rooting and hardening-off culture primary surface, it is 10 ± 1% to enable Rooting and hardening-off culture base water content
(volume ratio), in shading greenhouse it is lower place 2~3 days after, by the Rooting and hardening-off culture base of seedling base portion (refer in particular to this take root it is strong
Agar in seedling culture medium) it cleans, it dries to root system and whitens；
Finally by seedling with 3~5 for one clump transplanting in mixed-matrix in, cultivated in the greenhouse that shades；
It is described shading greenhouse light transmittance be 60 ± 5%, shading canopy temperature be 20~28 DEG C, relative humidity be 80 ±
5%；
The mixed-matrix is pine tree sawdust and pine bark according to 1:1 volume ratio mixing obtains.
Quick breeding by group culture further improvements in methods as dendrobium candidum of the present invention：
The method of clean capsule is in the step 1)：Capsule is placed in 1% liquid detergent solution (volume ratio) to shake and is washed
10min further takes out capsule and is rinsed well with tap water, drained and standby.
Compared with prior art, the present invention having following technical advantage：
1, the present invention sprouts differential medium using specific seed, seed sprout to be formed protocorm and protocorm differentiation at
Seedling carries out on same culture medium, and intermediate need not transfer replaces culture medium.
And conventional Seeds of Dendrobium Candidum culture, protocorm induction and differentiation separately carry out, intermediate demand to protocorm into
Culture medium is replaced in the primary switching of row, is manually expended very big.
2, added hormone kind is few in used each culture medium during tissue culture of the present invention and concentration is low, outer without addition
Source organic matter, the production is simple and convenient for culture medium, and tissue-cultured seedling production cost is low.
3, dendrobium candidum of the present invention can be transplanted from aseptic seeding to seedling about needs 230-240 days, and the production cycle is relatively existing
There is technology (production cycle is about 270-300 days) shorter.
4, the present invention carries out seeling industry using Seeds of Dendrobium Candidum, and breeding coefficient is high, and reproduction speed is fast, letter easy to operate
Just, human and material resources are saved, convenient for being factory produced.
5, Seeds of Dendrobium Candidum quick breeding by group culture method of the invention, Seeds of Dendrobium Candidum amount is big, and culture medium is simple for production,
Tissue-cultured seedling production cost is low, and the high quality seedling that a large amount of genetic backgrounds are identical, growing way is consistent can be produced within the relatively short period
(seed), and it is provided for a long term high quality seedling by what laboratory may be implemented the anniversary, it is particularly suitable for dendrobium candidum batch production
Nursery.Using the present invention, can simultaneously be effectively coyoting, shape are dug in this rare wild plant resource of protection dendrobium candidum, reduction private
At the mutually coordinated benign cycle of sustainable use and reasonable development.
Description of the drawings
The specific implementation mode of the present invention is described in further detail below in conjunction with the accompanying drawings.
Fig. 1 is the dendrobium candidum capsule that artificial pollination is tied；
Fig. 2 is that seed sprouts the seedling being differentiated to form in embodiment 1；
Fig. 3 be cultivated in seedling proliferation culture medium in embodiment 2 seedling (seedling proliferation culture medium be MS+
NAA0.2mg/L+ sucrose 30g/L+ agar 7g/L+0.5g/L activated carbons, pH 5.8)；
Fig. 4 is the tufted seedling cultivated in strengthening seedling and rooting culture medium in embodiment 1.
Fig. 5 is the seedling for intending transplanting in embodiment 1；
Fig. 6 is the seedling that one month is grown after being transplanted in embodiment 1.
Specific implementation mode
With reference to specific embodiment, the present invention is described further, but protection scope of the present invention is not limited in
This.
The seed group fast tissue culture reproducing method of embodiment 1, a kind of dendrobium candidum, follows the steps below successively：
1) it, draws materials：
As shown in Figure 1, taking ripe capsule (150 days fruit ages that artificial pollination is tied on the dendrobium candidum plant of robust growth
Do not crack above and), it places it in 1% liquid detergent solution (volume ratio) to shake and washes 10min, further take out capsule and rinsed with tap water
Totally, drained and standby.
2), the induction of seed is sprouted and is broken up：
Capsule step 1) is cleaned, drained is after routine disinfection (i.e. first with 75% alcohol immersion treatment 2min, sterile water
After flushing, sterilized 10min with 0.1% mercuric chloride solution, aseptic filter paper suck dry moisture used after aseptic water washing 3~5 times), excision is first
It is longitudinally splitted after each 0.5cm in tail both ends, takes out seed, and seed is uniformly inoculated on seed sprouting differential medium and is cultivated.
Condition of culture is：Black cloth covering first carries out light culture 10 days, and cultivation temperature is 25 ± 1 DEG C of constant temperature.
Then black cloth is thrown off, daily illumination in 14 hours, 30~40 μm of ol m-2s-1 of intensity of illumination, temperature is 25 ± 1
℃；10 hours light cultures, temperature are 21 ± 1 DEG C；Above-mentioned illumination and light culture are alternately.
Seed sprouts differential medium：MS+NAA0.1mg/L+ sucrose 30g/L+ agar 7g/L+ activated carbons 0.5g/L, pH
It is 5.8.
About 30 days or so, seed sprouted greening and forms protocorm, which sprouts in same seed in differential medium
It is differentiated to form seedling again.Seed sprouts differentiation rate and is up to 99%, and differentiation speed is fast and seedling is neat (as shown in Figure 2).
3), the proliferation of seedling：
(seed sprouts differentiation culture about 75-80 when seed is sprouted and is differentiated to form the unrooted seedling with two panels leaflet
It), Multiplying culture will be carried out on unrooted seedling inoculation to seedling proliferation culture medium, to obtain tufted seedling.
Condition of culture is：Illumination in 16 hours, 30~40 μm of ol m of intensity of illumination-2·s-1, temperature is 25 ± 1 DEG C；8 hours
Light culture, temperature are 21 ± 1 DEG C；Above-mentioned illumination and light culture are alternately；
Seedling proliferation culture medium is：MS+NAA0.15mg/L+ sucrose 30g/L+ agar 7g/L+0.5g/L activated carbons, pH are
5.8。
Seedling starts to be proliferated after about 15 days, and statistics seedling average proliferation multiple is 3.5 within the 90th day, and tufted seedling is elongated, height of seedling
2-3cm has short 3-4 item.
Remarks explanation：It, can be by seedling weight more sturdy in tufted seedling obtained by above-mentioned proliferation in order to obtain more tufted seedlings
The breeding of step 3) is carried out again.
4), strong plantlets and rootage：
Wait for that the tufted seedling that Multiplying culture obtains is grown to 2-3cm or so Gao Shi (seedling proliferation culture about 90-95 days), it will be above-mentioned
Tufted seedling is split to form single plant seedling from base portion, is inoculated on Rooting and hardening-off culture base and carries out Rooting and hardening-off culture (such as Fig. 4
It is shown)；
Condition of culture is：Illumination in 16 hours, 30~40 μm of ol m of intensity of illumination-2·s-1, temperature is 25 ± 1 DEG C；8 hours
Light culture, temperature are 21 ± 1 DEG C；Above-mentioned illumination and light culture are alternately；
Rooting and hardening-off culture base is：MS+BA0.5mg/L+NAA0.5mg/L+ sucrose 30g/L+ agar 8g/L+0.5g/L lives
Property charcoal, pH 5.8.
Seedling starts to take root within about 20 days, and statistics rooting rate is 100% within the 60th day.
5) it, transplants：
The seedling that under growth root strong seedling culture obtains is grown to 4.0~5.0cm high and grows root of the length more than or equal to 2cm extremely
At few 2 (Rooting and hardening-off culture about 60 days), obtain can bottle outlet plantation seedling (as shown in Figure 5)；
The seedling bottle outlet plantation method be specially：
First by gained can the seedling of bottle outlet plantation be transferred to culture vessel the shading greenhouse (light transmittance of quasi- rooting culture
For hardening 5~7 days (temperature is 20~28 DEG C) in 60%)；
Then bottle cap is opened, one layer of water (water content is 10% (volume ratio)) is poured in Rooting and hardening-off culture primary surface, then at
Shade after being placed 2-3 days in greenhouse (temperature be 20~28 DEG C), by the Rooting and hardening-off culture base of seedling base portion (refer in particular to this take root it is strong
Agar in seedling culture medium) it cleans, it dries to root system and whitens；
Finally seedling is transplanted for one clump in pine tree sawdust with 3~5：Pine bark (volume ratio) is 1:1 mixed base
It in matter, is cultivated in shading greenhouse (light transmittance 60%), cultivation temperature is 20~28 DEG C.
Dendrobium candidum of the present invention can be transplanted from aseptic seeding to seedling about needs 230-240 days, during entire transplanting,
Relative humidity keeps 80% or so in control shading greenhouse.
One month seedling is grown after the present embodiment transplanting as shown in fig. 6, its transplanting survival rate is up to 95% or more.
NAA concentration in 1 step 3) seedling proliferation culture medium of embodiment is changed to by embodiment 2 by " 0.15mg/L "
" 0.2mg/L ", remaining is equal to embodiment 1.
As shown in figure 3, unrooted seedling is proliferated in the seedling proliferation culture medium of a concentration of 0.2mg/L of NAA.
The proliferation results of seedling are：The tufted seedling that can further transfer is obtained after 90-95 days, statistics seedling is average within the 90th day
Proliferation times are 3.8, and tufted seedling is sturdy, height of seedling 2-3cm, there is short 2-3 item.
It waits for that above-mentioned gained tufted seedling is grown to 2-3cm or so Gao Shi, it is subjected to strong plantlets and rootage training according to 1 step 4) of embodiment
It supports, seedling starts to take root within about 22 days, and statistics rooting rate is 100% within the 60th day.
Comparative example 1, the minimal medium that the seed in 1 step 2) of embodiment is sprouted to differential medium are changed to by " MS "
" 1/2MS ", that is, seed sprouts differential medium and is：1/2MS minimal medium+NAA0.1mg/L+ sucrose 30g/L+ agar 7g/L
+ 0.5g/L activated carbons, pH 5.8, remaining is equal to embodiment 1.
Seed sprouts differentiated result：Seed sprouts differentiation rate and there was only 72%, is differentiated to form the unrooted with two panels leaflet
(can be transplanted from aseptic seeding to seedling and about need 260~270 days) slow-growing after seedling.
NAA concentration in seed sprouting differential medium in 1 step 2) of embodiment is changed to by comparative example 2 by 0.1mg/L
0.2mg/L, pH 5.8, remaining is equal to embodiment 1.
Seed sprouts differentiated result：Seed sprout differentiation rate there was only 48%, Some seeds are sprouted to form protocorm after become
In vain, stop differentiation.The growth of seedling being differentiated to form is very fast.
NAA concentration in seed sprouting differential medium in 1 step 2) of embodiment is changed to be changed by 0.1mg/L by comparative example 3
For 0.5mg/L, pH 5.8, remaining is equal to embodiment 1.
Seed sprouts differentiated result：Only 60% seed can induce sprouting and form protocorm, and 95% protocorm
Later stage bleaches, and stops differentiation, and seed sprouts differentiation rate and there was only 3%.
Comparative example 4 sprouts addition BA1.0mg/L, mashed potato 20g/ in differential medium in 1 step 2) seed of embodiment
L, that is, seed sprouts differential medium and is：MS+NAA0.1mg/L+BA1.0mg/L+ sucrose 30g/L+ agar 7g/L+ activated carbons
0.5g/L+ mashed potato 20g/L, remaining is equal to embodiment 1.
Seed sprouts differentiated result：Seed sprout differentiation rate there was only 20%, most of seed is sprouted to form protocorm after
Bleach, stops differentiation.
NAA concentration in 1 step 3) seedling proliferation culture medium of embodiment is changed to 0.5mg/ by comparative example 5 by 0.15mg/L
L, remaining is equal to embodiment 1.
The proliferation results of seedling are：The tufted seedling that can further transfer is obtained after 90-95 days, statistics seedling is average within the 90th day
Proliferation times are 2.5, and tufted seedling is sturdy, height of seedling 1-2cm, unrooted.
Comparative example 6 adds mashed potato 20g/L in 1 step 3) seedling proliferation culture medium of embodiment, that is, seedling proliferation is trained
Foster base is：MS+NAA0.15mg/L+ sucrose 30g/L+ agar 7g/L+0.5g/L activated carbons+mashed potato 20g/L, remaining is equivalent
In embodiment 1.
The proliferation results of seedling are：The tufted seedling that can further transfer is obtained after 90-95 days, statistics seedling is average within the 90th day
Proliferation times are 2.8, and tufted seedling growth potential is weak, height of seedling 1-2cm, unrooted.
Finally, it should also be noted that it is listed above be only the present invention several specific embodiments.Obviously, this hair
Bright to be not limited to above example, acceptable there are many deformations.Those skilled in the art can be from present disclosure
All deformations for directly exporting or associating, are considered as protection scope of the present invention.
Claims (6)
1. the seed group fast tissue culture reproducing method of dendrobium candidum, it is characterized in that including the following steps successively：
1) it, draws materials：
150 days fruit ages tying above uncracked maturation capsule on the dendrobium candidum of robust growth is selected, after being cleaned, draining
It is spare；
2), the induction of seed is sprouted and is broken up：
By the capsule obtained by step 1) it is sterilized after, longitudinally splitted after cutting off each 0.3~0.7cm in its head and the tail both ends, take out seed,
Seed is uniformly inoculated on seed sprouting differential medium and is cultivated；
Condition of culture is：
Light culture is carried out first 10 ± 1 days, temperature is 25 ± 1 DEG C；
Carry out illumination in 14 hours, 30~40 μm of ol m of intensity of illumination daily later-2·s-1, temperature is 25 ± 1 DEG C；Dark training in 10 hours
It supports, temperature is 21 ± 1 DEG C；Above-mentioned illumination and light culture are alternately；
3), the proliferation of seedling：
When seed is sprouted and is differentiated to form the seedling of two panels leaflet, by the switching of the seedling of gained in seedling proliferation culture medium into
Row Multiplying culture obtains tufted seedling；
Condition of culture is：Illumination in 16 hours, 30~40 μm of olm of intensity of illumination-2s-1, temperature is 25 ± 1 DEG C；8 hours light cultures, temperature
Degree is 21 ± 1 DEG C；Above-mentioned illumination and light culture are alternately；
4), strong plantlets and rootage：
It when the tufted seedling that Multiplying culture obtains is grown to 2-3cm high, is split tufted seedling to form single plant seedling from base portion, turn
It moves on to and carries out Rooting and hardening-off culture on Rooting and hardening-off culture base；
Condition of culture is：Illumination in 16 hours, 30~40 μm of oLm of intensity of illumination-2s-1, temperature is 25 ± 1 DEG C；8 hours light cultures, temperature
Degree is 21 ± 1 DEG C；Above-mentioned illumination and light culture are alternately；
5) it, transplants：
Under growth root strong seedling culture obtain seedling grow to 4.0~5.0cm high and grow length more than or equal to 2cm root at least 2
When, acquisition can bottle outlet plantation seedling.
2. the seed group fast tissue culture reproducing method of dendrobium candidum according to claim 1, it is characterized in that：
Seed in the step 2) sprouts differential medium：MS+NAA0.05~0.15mg/L+ sucrose 25~35g/L+ fine jades
5~10g/L+0.3 of fat~0.7g/L activated carbons, pH are 5.5~6.
3. the seed group fast tissue culture reproducing method of dendrobium candidum according to claim 2, it is characterized in that：
Seedling proliferation culture medium in step 3) is：5~10g/L of MS+NAA0.15~0.2mg/L+ sucrose 25~35g/L+ agar
+ activated carbon 0.3~0.7g/L, pH are 5.5~6.
4. the seed group fast tissue culture reproducing method of dendrobium candidum according to claim 3, it is characterized in that：
Rooting and hardening-off culture base in step 4) is MS+BA0.3~0.7mg/L+NAA0.3~25~35g/L of 0.7mg/L+ sucrose
+ agar 5~10g/L+ activated carbons 0.3~0.7g/L, pH are 5.5~6.
5. the seed group fast tissue culture reproducing method of dendrobium candidum according to any one of claims 1 to 4, it is characterized in that：
First by gained can bottle outlet plantation seedling hardening 5~7 in the shading greenhouse of quasi- rooting culture is transferred to culture vessel
It；
Then bottle cap is opened, is watered in Rooting and hardening-off culture primary surface, it is 10 ± 1% to enable Rooting and hardening-off culture base water content, then
After being placed 2~3 days under in the greenhouse that shades, the Rooting and hardening-off culture base of seedling base portion is cleaned, dries to root system and whitens；
Finally by seedling with 3~5 for one clump transplanting in mixed-matrix in, cultivated in the greenhouse that shades；
The light transmittance of the shading greenhouse is 60 ± 5%, and shading canopy temperature is 20~28 DEG C, and relative humidity is 80 ± 5%；
The mixed-matrix is pine tree sawdust and pine bark according to 1:1 volume ratio mixing obtains.
6. the seed group fast tissue culture reproducing method of dendrobium candidum according to claim 5, it is characterized in that：
The method of clean capsule is in the step 1)：Capsule is placed in 1% liquid detergent solution to shake and washes 10min, further takes out capsule
Fruit is rinsed well with tap water, drained and standby.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810909426.7A CN108617511A (en) | 2018-08-10 | 2018-08-10 | The seed group fast tissue culture reproducing method of dendrobium candidum |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810909426.7A CN108617511A (en) | 2018-08-10 | 2018-08-10 | The seed group fast tissue culture reproducing method of dendrobium candidum |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108617511A true CN108617511A (en) | 2018-10-09 |
Family
ID=63708887
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810909426.7A Pending CN108617511A (en) | 2018-08-10 | 2018-08-10 | The seed group fast tissue culture reproducing method of dendrobium candidum |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108617511A (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110100737A (en) * | 2019-06-21 | 2019-08-09 | 台州市椒江草之堂生物科技有限公司 | A kind of candidum tissue culturing rapid propagation method |
| CN110291987A (en) * | 2019-06-20 | 2019-10-01 | 成都师范学院 | A kind of Seeds of Dendrobium Candidum culture medium adding natural phytohormone |
| CN112237140A (en) * | 2020-10-16 | 2021-01-19 | 云南省农业科学院花卉研究所 | Method for blocking dendrobium officinale tissue culture seedling from being polluted by bacteria |
| CN112273232A (en) * | 2020-10-31 | 2021-01-29 | 浙江省农业科学院 | Curcuma wenyujin detoxification seedling culture method |
| CN114303950A (en) * | 2021-12-31 | 2022-04-12 | 上海应用技术大学 | Tissue culture and rapid propagation method for dendrobium officinale seeds |
| CN115804342A (en) * | 2022-12-05 | 2023-03-17 | 贵州康旅食斛开发有限公司 | Tissue culture method for dendrobium officinale |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104429961A (en) * | 2014-12-08 | 2015-03-25 | 河源市裕森农林发展有限公司 | Rapid propagation tissue culture method for dendrobium officinale |
| CN105052753A (en) * | 2015-09-29 | 2015-11-18 | 江苏农林职业技术学院 | Rapid propagation method for dendrobii officmalis caulis |
-
2018
- 2018-08-10 CN CN201810909426.7A patent/CN108617511A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104429961A (en) * | 2014-12-08 | 2015-03-25 | 河源市裕森农林发展有限公司 | Rapid propagation tissue culture method for dendrobium officinale |
| CN105052753A (en) * | 2015-09-29 | 2015-11-18 | 江苏农林职业技术学院 | Rapid propagation method for dendrobii officmalis caulis |
Non-Patent Citations (3)
| Title |
|---|
| 石丽敏等: "铁皮石斛原球茎诱导分化与增殖试验", 《浙江农业科学》 * |
| 石丽敏等: "铁皮石斛栽培基质筛选研究", 《农业科技通讯》 * |
| 许奕等: "不同培养基对铁皮石斛壮苗生根的影响及移栽条件优化", 《江苏农业科学》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110291987A (en) * | 2019-06-20 | 2019-10-01 | 成都师范学院 | A kind of Seeds of Dendrobium Candidum culture medium adding natural phytohormone |
| CN110100737A (en) * | 2019-06-21 | 2019-08-09 | 台州市椒江草之堂生物科技有限公司 | A kind of candidum tissue culturing rapid propagation method |
| CN112237140A (en) * | 2020-10-16 | 2021-01-19 | 云南省农业科学院花卉研究所 | Method for blocking dendrobium officinale tissue culture seedling from being polluted by bacteria |
| CN112273232A (en) * | 2020-10-31 | 2021-01-29 | 浙江省农业科学院 | Curcuma wenyujin detoxification seedling culture method |
| CN114303950A (en) * | 2021-12-31 | 2022-04-12 | 上海应用技术大学 | Tissue culture and rapid propagation method for dendrobium officinale seeds |
| CN115804342A (en) * | 2022-12-05 | 2023-03-17 | 贵州康旅食斛开发有限公司 | Tissue culture method for dendrobium officinale |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108617511A (en) | The seed group fast tissue culture reproducing method of dendrobium candidum | |
| CN102301953B (en) | Method for efficiently and rapidly propagating Dendrobium candidum test-tube seedlings in large scale | |
| CN102204512B (en) | Tissue culture method for lilium tenuifolium | |
| CN101116424B (en) | Highly effective lily bulblet inducement culture method | |
| CN104904597A (en) | Bletilla striata rapid propagation and sunshine seedling strengthening method | |
| CN104871679A (en) | Balance dormancy breaking and seedling cultivation method of cerasus campanulata seeds | |
| CN107197746A (en) | A kind of mating system of China fir field excellent resources | |
| CN106922536A (en) | A kind of method that fast energy-saving cultivates bletilla seedling | |
| CN108040879B (en) | Limonium aureum rooting induction culture medium and Limonium aureum breeding method | |
| CN104823846B (en) | The method for quickly breeding of Zhejiang Anoectochilus nefiliforme (Nakai) hara seedling | |
| CN106942053B (en) | A kind of tissue culture and rapid propagation method for Xingan lucid asparagus | |
| CN108782247A (en) | A kind of method for tissue culture of late cherry " Yu Yihuang " kind of Japan | |
| CN106857258A (en) | A kind of quick breeding method for tissue culture blue with leaf pocket | |
| CN106386504A (en) | Tissue culture method of Aralia Cordata Thunb seedlings | |
| CN105010123B (en) | The method and culture medium of strawberry distant hybrid are obtained by rescue isolated culture | |
| CN110402818A (en) | A kind of method of excellent kind of Chinese chestnut Mature Embryo Tissue culture rapid seedling cultivation | |
| CN107006367B (en) | 'sunshine' cherry tissue culture rapid propagation method | |
| CN108522279A (en) | The direct seedling tissue culture method of wild rice stem seed embryo and used medium | |
| CN108260529A (en) | A kind of tissue cultivating and seedling method of red root wild silkworm beans | |
| CN105766636B (en) | A kind of peony tissue culture regeneration method | |
| CN104620983A (en) | Method for forming seedlings by carrying out tissue culture on echinochloacrus-galli | |
| CN107646694A (en) | A kind of adenophora tetraphylla Regeneration in Vitro tissue cultures technology | |
| CN109258463B (en) | Vegetative propagation method of paphiopedilum armeniacum | |
| CN104221851B (en) | A kind of great Ye ant tower isolated culture and rapid propagation method | |
| CN110100737A (en) | A kind of candidum tissue culturing rapid propagation method |
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 |
Application publication date: 20181009 |
|
| RJ01 | Rejection of invention patent application after publication |