CN106172008A - A kind of Bowring cattleya fast culture propagation method - Google Patents
A kind of Bowring cattleya fast culture propagation method Download PDFInfo
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- CN106172008A CN106172008A CN201610661975.8A CN201610661975A CN106172008A CN 106172008 A CN106172008 A CN 106172008A CN 201610661975 A CN201610661975 A CN 201610661975A CN 106172008 A CN106172008 A CN 106172008A
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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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Abstract
The present invention relates to technical field of plant culture, it is specifically related to a kind of Bowring cattleya fast culture propagation method, comprises the following steps: material selection, outer implant pretreatment, the sterilization of outer implant, inducing culture, ultrasonic Treatment, enrichment culture, root culture, strong seedling culture and transplanting.Bowring cattleya can be carried out fast culture breeding by the present invention; efficiency is high; and the Bowring cattleya Seedling strain planting percent after Pei Yanging is high; Seedling strain quality better; strong adaptability after cultivation; by animal nutrition at short notice, meet the demand commercially produced, cultivate Bowring cattleya provide technical support for enterprise scale, industrialization.This tissue culture propagation method Disinfection Effect is good simultaneously, browning rate is low, thus substantially increases planting percent and work efficiency, has saved resource.
Description
Technical field
The present invention relates to technical field of plant culture, be specifically related to a kind of Bowring cattleya fast culture propagation method.
Background technology
Bowring cattleya, is one of the most most notable Cymbidium ensifolium (L.) Sw..Originate in american torrid zone, for state's national flowers such as Brazil, Colombia.
Kind is more than thousands of, and color has white, yellow, green, purple etc..Breeding plant division, tissue culture or aseptic seeding, property happiness temperature
Warm, moist and sufficient illumination.The Main Means that Bowring cattleya is efficiently bred is exactly tissue culture.More external developed countries are usually
Use industrial breeding technique and tissue culture rapid propagating technology, be possible not only to create huge economic benefit, also cultivate simultaneously
A lot of improved seeds, but China studies about the fast breeding technique of Bowring cattleya tissue culture and is still within the primary stage, very
The large-scale production of Bowring cattleya is hindered in big degree.And the tissue culture of other cymbidium varieties existing can not directly use
Cultivate to Bowring cattleya, so being badly in need of a kind of Bowring cattleya fast culture propagation method the most at present.
Summary of the invention
It is an object of the invention to provide a seedling strain quality better, cultivate the Bowring cattleya fast culture breeding side that breeding is fast
Method.
In order to achieve the above object, the technical solution used in the present invention is: a kind of Bowring cattleya fast culture propagation method, bag
Include following steps:
(1) material selection: choose the Bowring cattleya plant grown fine without pest and disease damage, carry out the shading treatment of 5～8d, shading
After process, choose the sprouting on growing way stalwartness plant as outer implant；
(2) outer implant pretreatment: by the outer implant bud wound surface of antioxidant solution washing just cutting；
(3) outer implant sterilization: tentatively cleaned in pond by the outer implant bud chosen with soft bristle, removes surface
Silt, then clean up with liquid detergent, be placed under flowing water flushing 30min, on super-clean bench, then divest outer implant bud
Outmost one layer of bract, soaks 1～2s, then at the hypochlorous acid being immersed in 10% in 75% ethanol crossed through high temperature sterilize
Initial sterilization 10min in sodium solution, after initial sterilization, with aseptic water washing 5 times, then the bract that peels a layer from, put into the secondary chlorine of 5%
Sterilize in acid sodium solution 5min again, and then using the mode of repeated disinfection, the concentration of each sodium hypochlorite is sterilization last time
Half, the half that disinfecting time was also sterilized for last time, until stripping is to leaving last sprout, finally cut the bud point of about 5mm,
Bubble is sterilized 1min in the liquor natrii hypochloritis of 0.5%, with aseptic water washing 6～8 times, after blotting surface moisture with sterilized filter paper
It is positioned in culture dish standby；
(4) inducing culture: by the outer implant after sterilization, be seeded in inducing culture and cultivate, first carry out 7～10d
Low-light cultivate, intensity of illumination is 500～700lx, is then carrying out conventional illumination cultivation, intensity of illumination be 1500～
2000lx, cultivation temperature is 24～26 DEG C, and light application time is 13h/d；Described inducing culture is: 1/2MS+0.4～0.8mg/L
6-BA+0.05～0.08mg/L NAA+10～15g/L sucrose+6～8g/L agar+10～15% coconut juice+490～510mg/L lemon
Lemon acid+5～7g/L PVP, pH value is 5.4～5.5；
(5) ultrasonic Treatment: the protocorm induced is placed in the ultrasound wave that frequency is 25kHz process 3～4min；
(6) enrichment culture: being transferred to after ultrasonic Treatment in proliferated culture medium continue to cultivate, described proliferated culture medium is:
1/2MS+1～2mg/L 6-BA+0.4～0.6mg/L NAA+10～15g/L sucrose+6～8g/L agar+10～15% coconut juice+
490～510mg/L citric acid+2～4g/L PVP, pH value is 5.4～5.5；Cultivation temperature 25～28 DEG C, light application time is 13h/
D, intensity of illumination is 1500～2000lx；
(7) root culture: after enrichment culture, proceeds to cultivate in root media, and described root media is: 1/2MS+
0.1～0.3mg/L NAA+140～160g/L banana puree+90～110mg/L citric acid+28～32g/L sugar+5～9g/L fine jade
Fat, pH is 5.4～5.5, cultivation temperature 25～28 DEG C, and light application time is 13h/d, and intensity of illumination is 1500～2000lx；
(8) strong seedling culture: select the aseptic seedling of high more than 3cm to proceed in strong seedling culture base and cultivate, described strong seedling culture base
For: 1/2MS+0.3～0.7mg/L 6-BA+0.1～0.2mg/L GA3+10～20g/L sucrose+10～20mg/L glucose+
190～200g/L banana puree+4～6g/L agar, pH is 5.4～5.5, cultivation temperature 25～28 DEG C, and light application time is 13h/d,
Intensity of illumination is 1500～2000lx；
(9) transplant: when test tube Seedling grows to 5～7cm high, leaf 2～when 3, test tube Seedling is placed on nature light lower refining seedling 5～
7 days, open bottle cap seedling exercising 1～2 days；Then take out cultivation Seedling, clean the culture medium being attached to root with clear water, note during cleaning
Not damaging root, then transplant to culture matrix, keep humidity and ventilation, air humidity is 75～85%, and temperature is 20
～25 DEG C.
Bowring cattleya fast culture propagation method as above a kind of, further illustrate into, described antioxidant solution selects
With glutathion or ascorbic acid.
Bowring cattleya fast culture propagation method as above a kind of, further illustrate into, described inducing culture is: 1/
2MS+0.6mg/L 6-BA+0.07mg/L NAA+13g/L sucrose+7g/L agar+13% coconut juice+500mg/L citric acid+6g/L
PVP。
Bowring cattleya fast culture propagation method as above a kind of, further illustrate into, described proliferated culture medium is: 1/
2MS+1.5mg/L 6-BA+0.5mg/L NAA+13g/L sucrose+7g/L agar+13% coconut juice+500mg/L citric acid+3g/L
PVP。
Bowring cattleya fast culture propagation method as above a kind of, further illustrate into, described root media is: 1/
2MS+0.2mg/L NAA+150g/L banana puree+100mg/L citric acid+30g/L sugar+7g/L agar.
Bowring cattleya fast culture propagation method as above a kind of, further illustrate into, described strong seedling culture base is: 1/
2MS+0.5mg/L 6-BA+0.15mg/L GA3+15g/L sucrose+15mg/L glucose+195g/L banana puree+5g/L agar.
Bowring cattleya fast culture propagation method as above a kind of, further illustrate into, in described strong seedling culture step,
Heavy caliber Cymbidium ensifolium (L.) Sw. vial is used to carry out Aseptic seedling culture.
The invention has the beneficial effects as follows: by the present invention, Bowring cattleya can carry out fast culture breeding, efficiency is high, and trains
Bowring cattleya Seedling strain planting percent after Yanging is high, and Seedling strain quality better, strong adaptability after cultivation, by animal nutrition in the short time
In, meet the demand commercially produced, cultivate Bowring cattleya provide technical support for enterprise scale, industrialization.The most originally
Tissue culture propagation method Disinfection Effect is good, browning rate is low, thus substantially increases planting percent and work efficiency, has saved resource.
Accompanying drawing explanation
Fig. 1 is schematic flow sheet of the present invention.
Detailed description of the invention
Below in conjunction with the accompanying drawings embodiment of the present invention is further elaborated.
As it is shown in figure 1, a kind of Bowring cattleya fast culture propagation method that the present invention provides, comprise the following steps: material selects
Take, outer implant pretreatment, the sterilization of outer implant, inducing culture, ultrasonic Treatment, enrichment culture, root culture, strong seedling culture and shifting
Planting, this method Disinfection Effect is good, it is fast to cultivate breeding, planting percent is high and browning rate is low such that it is able to reduces Bowring cattleya and trained in group
Browning rate in journey, increases planting percent, improves work efficiency, elaborates each step in method below.
(1) material selection: choose the Bowring cattleya plant grown fine without pest and disease damage, carry out the shading treatment of 5～8d, shading
After process, choose the sprouting on growing way stalwartness plant as outer implant；Use the Bowring cattleya plant energy grown fine without pest and disease damage
Enough reduce the sickness rate of Seedling strain, it is ensured that do not disturbed by other factors during group training, increase outer implant survival rate, and warp
Cross shading treatment, it is possible to reduce browning rate.
(2) outer implant pretreatment: by the outer implant bud wound surface of antioxidant solution washing just cutting；Can be maximum
Degree ground reduces the chance that contact with outer implant tangent plane of oxygen, the most sufficiently in the outer implant of precipitation water miscible polyphenol substance with
Quinones substance, the oxidation of less aldehydes matter, thus effectively alleviate the outer implant brown stain in tissue culture procedures of Bowring cattleya,
As preferably, glutathione solution selected by described antioxidant, it is also possible to select ascorbic acid, it is also possible to by several antioxidants
It is used in conjunction with.
(3) outer implant sterilization: tentatively cleaned in pond by the outer implant bud chosen with soft bristle, removes surface
Silt, then clean up with liquid detergent, be placed under flowing water flushing 30min, on super-clean bench, then divest outer implant bud
Outmost one layer of bract, soaks 1～2s, then at the hypochlorous acid being immersed in 10% in 75% ethanol crossed through high temperature sterilize
Initial sterilization 10min in sodium solution, after initial sterilization, with aseptic water washing 5 times, then the bract that peels a layer from, put into the secondary chlorine of 5%
Sterilize in acid sodium solution 5min again, and then using the mode of repeated disinfection, the concentration of each sodium hypochlorite is sterilization last time
Half, disinfecting time is also for the half of sterilization last time, until stripping is to leaving last sprout, the most next with 2.5% secondary
Sodium chlorate solution sterilizes 2.5min again, and peel a layer from after flushing bract, more again sterilizes with the liquor natrii hypochloritis of 1.25%
1.25min, the like；Finally cutting the bud point of about 5mm, bubble is sterilization 1min in the liquor natrii hypochloritis of 0.5%, by nothing
Bacterium water rinses 6～8 times, is positioned in culture dish standby after blotting surface moisture with sterilized filter paper.Use staged sterilization,
While constantly peelling off bud clothing, use the liquor natrii hypochloritis of variable concentrations to carry out disinfection, Disinfection Effect can be made more preferable, reduce
While germ contamination, improve survival rate.
(4) inducing culture: by the outer implant after sterilization, be seeded in inducing culture and cultivate, first carry out 7～10d
Low-light cultivate, intensity of illumination is 500～700lx, is then carrying out conventional illumination cultivation, intensity of illumination be 1500～
2000lx, cultivation temperature is 24～26 DEG C, and light application time is 13h/d；Table 1 is the impact using low-light to cultivate external implant:
Being placed on above-mentioned two groups in identical culture medium and cultivate, condition of culture is all consistent, simply uses low-light
One group cultivated starts first 8 days first to carry out low-light cultivation, and intensity of illumination is 500～700lx, does not use low-light to cultivate
One group directly carries out conventional illumination cultivation, and intensity of illumination is 1500～2000lx, the most first carries out a period of time
Low-light cultivate, it is possible to make outer implant gradually adapt to condition of culture in incubation, be conducive to survival, Restrain browning simultaneously
Generation.
Inducing culture described in this step is: 1/2MS+0.4～0.8mg/L 6-BA+0.05～0.08mg/L NAA+
10～15g/L sucrose+6～8g/L agar+10～15% coconut juice+490～510mg/L citric acid+5～7g/L PVP, pH value is
5.4～5.5；Table 2 is the various embodiments of inducing culture:
As preferably, described inducing culture is: 1/2MS+0.6mg/L 6-BA+0.07mg/L NAA+13g/L sucrose+
7g/L agar+13% coconut juice+500mg/L citric acid+6g/L PVP, is composition and the consumption of culture medium 1 in table 2.
(5) ultrasonic Treatment: the protocorm induced is placed in the ultrasound wave that frequency is 25kHz process 3～4min；Logical
After crossing ultrasonic Treatment, the appreciation rate of protocorm is higher by 45% than the protocorm not carrying out any process, thus improves planting percent,
Accelerate the proliferative speed of Bowring cattleya group training, substantially increase work efficiency, provide skill for enterprise's rapid cultivation Bowring cattleya
Art is supported.
(6) enrichment culture: being transferred to after ultrasonic Treatment in proliferated culture medium continue to cultivate, described proliferated culture medium is:
1/2MS+1～2mg/L 6-BA+0.4～0.6mg/L NAA+10～15g/L sucrose+6～8g/L agar+10～15% coconut juice+
490～510mg/L citric acid+2～4g/L PVP, pH value is 5.4～5.5；Cultivation temperature 25～28 DEG C, light application time is 13h/
D, intensity of illumination is 1500～2000lx；Table 3 is the various embodiments of proliferated culture medium:
As preferably, described proliferated culture medium is: 1/2MS+1.5mg/L 6-BA+0.5mg/L NAA+13g/L sucrose+
7g/L agar+13% coconut juice+500mg/L citric acid+3g/L PVP, is composition and the consumption of culture medium 7 in table 3.
(7) root culture: after enrichment culture, proceeds to cultivate in root media, and described root media is: 1/2MS+
0.1～0.3mg/L NAA+140～160g/L banana puree+90～110mg/L citric acid+28～32g/L sugar+5～9g/L fine jade
Fat, pH is 5.4～5.5, cultivation temperature 25～28 DEG C, and light application time is 13h/d, and intensity of illumination is 1500～2000lx；Table 4 is
The various embodiments of root media:
As preferably, described root media is: 1/2MS+0.2mg/L NAA+150g/L banana puree+100mg/L Fructus Citri Limoniae
Acid+30g/L sugar+7g/L agar, is composition and the consumption of culture medium 13 in table 4.
(8) strong seedling culture: selecting the aseptic seedling of high more than 3cm to proceed in strong seedling culture base and cultivate, aseptic seedling root cap is the biggest,
More it is beneficial to Seedling strain and absorbs the nutrient substance in culture medium, promote Seedling strain growth, make Seedling strain upgrowth situation the best, therefore select high 3cm
Above aseptic seedling carries out next step cultivation.Described strong seedling culture base is: 1/2MS+0.3～0.7mg/L 6-BA+0.1～
0.2mg/L GA3+10～20g/L sucrose+10～20mg/L glucose+190～200g/L banana puree+4～6g/L agar, pH is
5.4～5.5, cultivation temperature 25～28 DEG C, light application time is 13h/d, and intensity of illumination is 1500～2000lx；Table 5 was trained for strong sprout
The various embodiments of foster base:
As preferably, described strong seedling culture base is: 1/2MS+0.5mg/L 6-BA+0.15mg/L GA3+15g/L sucrose+
15mg/L glucose+195g/L banana puree+5g/L agar, is composition and the consumption of culture medium 19 in table 5.In this step,
Heavy caliber Cymbidium ensifolium (L.) Sw. vial being preferably used and carries out Aseptic seedling culture, table 6 is the impact that Bowring cattleya is grown by different culture vessel:
| Container | Seedling plant height degree (cm) | Root/shoot ratio | Growth conditions |
| Heavy caliber Cymbidium ensifolium (L.) Sw. vial | 4.51 | 0.690 | +++ |
| Small-bore Cymbidium ensifolium (L.) Sw. vial | 4.14 | 0.594 | ++ |
| Large-bore plastics bottle | 3.89 | 0.523 | ++ |
Seedling strain consistent for growing way being placed in same medium and cultivate, the most only container is inconsistent, unites after cultivating 30d
The result counted out, as can be seen from Table 6, uses heavy caliber Cymbidium ensifolium (L.) Sw. vial to carry out Aseptic seedling culture and is readily able to the life of Seedling strain
Long.
(9) transplant: when test tube Seedling grows to 5～7cm high, leaf 2～when 3, test tube Seedling is placed on nature light lower refining seedling 5～
7 days, open bottle cap seedling exercising 1～2 days；Then take out cultivation Seedling, clean the culture medium being attached to root with clear water, note during cleaning
Not damaging root, then transplant to culture matrix, keep humidity and ventilation, air humidity is 75～85%, and temperature is 20
～25 DEG C.By the present invention, Bowring cattleya can carry out fast culture breeding, efficiency is high, and the Bowring cattleya Seedling strain seedling after cultivation
Rate is high, Seedling strain quality better, strong adaptability after cultivation, by animal nutrition at short notice, meets and commercially produces
Demand, cultivates Bowring cattleya provide technical support for enterprise scale, industrialization.This tissue culture propagation method Disinfection Effect simultaneously
Good, browning rate is low, thus substantially increases planting percent and work efficiency, has saved resource.
The present invention is not limited to examples detailed above, in claims of the present invention limited range, and art technology
Various deformation or amendment that personnel can make without creative work are all protected by this patent.
Claims (7)
1. a Bowring cattleya fast culture propagation method, it is characterised in that comprise the following steps:
(1) material selection: choose the Bowring cattleya plant grown fine without pest and disease damage, carry out the shading treatment of 5～8d, shading treatment
After, choose the sprouting on growing way stalwartness plant as outer implant；
(2) outer implant pretreatment: by the outer implant bud wound surface of antioxidant solution washing just cutting；
(3) outer implant sterilization: tentatively cleaned in pond by the outer implant bud chosen with soft bristle, removes the mud on surface
Sand, then cleans up with liquid detergent, is placed under flowing water flushing 30min, then divests outer implant bud outermost on super-clean bench
One layer of bract in face, in 75% ethanol crossed through high temperature sterilize, immersion 1～2s, then molten at the sodium hypochlorite being immersed in 10%
Initial sterilization 10min in liquid, after initial sterilization, with aseptic water washing 5 times, then the bract that peels a layer from, put into the sodium hypochlorite of 5%
Sterilize in solution 5min again, the half then using the mode of repeated disinfection, the concentration of each sodium hypochlorite to be sterilization last time,
The half that disinfecting time was also sterilized for last time, until stripping is to leaving last sprout, finally cuts the bud point of about 5mm, steeps and exist
Sterilize in the liquor natrii hypochloritis of 0.5% 1min, with aseptic water washing 6～8 times, places after blotting surface moisture with sterilized filter paper
In culture dish standby；
(4) inducing culture: will sterilization after outer implant, be seeded in inducing culture and cultivate, first carry out 7～10d weak
Illumination cultivation, intensity of illumination is 500～700lx, is then carrying out conventional illumination cultivation, and intensity of illumination is 1500～2000lx,
Cultivation temperature is 24～26 DEG C, and light application time is 13h/d；Described inducing culture is: 1/2MS+0.4～0.8mg/L 6-BA+
0.05～0.08mg/L NAA+10～15g/L sucrose+6～8g/L agar+10～15% coconut juice+490～510mg/L citric acid+
5～7g/L PVP, pH value is 5.4～5.5；
(5) ultrasonic Treatment: the protocorm induced is placed in the ultrasound wave that frequency is 25kHz process 3～4min；
(6) enrichment culture: being transferred to after ultrasonic Treatment in proliferated culture medium continue to cultivate, described proliferated culture medium is: 1/2MS
+ 1～2mg/L 6-BA+0.4～0.6mg/L NAA+10～15g/L sucrose+6～8g/L agar+10～15% coconut juice+490～
510mg/L citric acid+2～4g/L PVP, pH value is 5.4～5.5；Cultivation temperature 25～28 DEG C, light application time is 13h/d, light
It is 1500～2000lx according to intensity；
(7) root culture: after enrichment culture, proceed in root media cultivate, described root media is: 1/2MS+0.1～
0.3mg/L NAA+140～160g/L banana puree+90～110mg/L citric acid+28～32g/L sugar+5～9g/L agar, pH
Being 5.4～5.5, cultivation temperature 25～28 DEG C, light application time is 13h/d, and intensity of illumination is 1500～2000lx；
(8) strong seedling culture: selecting the aseptic seedling of high more than 3cm to proceed in strong seedling culture base and cultivate, described strong seedling culture base is: 1/
2MS+0.3～0.7mg/L 6-BA+0.1～0.2mg/L GA3+10～20g/L sucrose+10～20mg/L glucose+190～
200g/L banana puree+4～6g/L agar, pH is 5.4～5.5, cultivation temperature 25～28 DEG C, and light application time is 13h/d, and illumination is strong
Degree is 1500～2000lx；
(9) transplant: when test tube Seedling grows to 5～7cm high, leaf 2～when 3, test tube Seedling be placed on nature light lower refining seedling 5～7 days,
Open bottle cap seedling exercising 1～2 days；Then take out cultivation Seedling, clean the culture medium being attached to root with clear water, be careful not to during cleaning
Infringement root, then transplants to culture matrix, keeps humidity and ventilation, and air humidity is 75～85%, and temperature is 20～25
℃。
A kind of Bowring cattleya fast culture propagation method the most according to claim 1, it is characterised in that: described antioxidant is molten
Liquid selects glutathion or ascorbic acid.
A kind of Bowring cattleya fast culture propagation method the most according to claim 1, it is characterised in that: described inducing culture
For: 1/2MS+0.6mg/L 6-BA+0.07mg/L NAA+13g/L sucrose+7g/L agar+13% coconut juice+500mg/L citric acid+
6g/L PVP。
A kind of Bowring cattleya fast culture propagation method the most according to claim 1, it is characterised in that: described proliferated culture medium
For: 1/2MS+1.5mg/L 6-BA+0.5mg/L NAA+13g/L sucrose+7g/L agar+13% coconut juice+500mg/L citric acid+
3g/L PVP。
A kind of Bowring cattleya fast culture propagation method the most according to claim 1, it is characterised in that: described root media
For: 1/2MS+0.2mg/L NAA+150g/L banana puree+100mg/L citric acid+30g/L sugar+7g/L agar.
A kind of Bowring cattleya fast culture propagation method the most according to claim 1, it is characterised in that: described strong seedling culture base
For: 1/2MS+0.5mg/L 6-BA+0.15mg/L GA3+15g/L sucrose+15mg/L glucose+195g/L banana puree+5g/L fine jade
Fat.
A kind of Bowring cattleya fast culture propagation method the most according to claim 1, it is characterised in that: described strong seedling culture walks
In Zhou, heavy caliber Cymbidium ensifolium (L.) Sw. vial is used to carry out Aseptic seedling culture.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106804425A (en) * | 2016-12-23 | 2017-06-09 | 中国林业科学研究院林业研究所 | A kind of cultural method of Bowring cattleya high-efficiency tissue culture |
| CN107018899A (en) * | 2017-04-21 | 2017-08-08 | 山东省农作物种质资源中心 | A kind of method of restoration ecosystem after Bowring cattleya germ plasm resource Plantlet in vitro and preservation |
| CN107278901A (en) * | 2017-07-31 | 2017-10-24 | 王生旭 | A kind of sterilization method of iris bennet bud |
| CN109601386A (en) * | 2019-01-24 | 2019-04-12 | 北京农业职业学院 | The sterilization method of bud explant is cut in a kind of state orchid stem tip tissue culture |
-
2016
- 2016-08-12 CN CN201610661975.8A patent/CN106172008A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106804425A (en) * | 2016-12-23 | 2017-06-09 | 中国林业科学研究院林业研究所 | A kind of cultural method of Bowring cattleya high-efficiency tissue culture |
| CN107018899A (en) * | 2017-04-21 | 2017-08-08 | 山东省农作物种质资源中心 | A kind of method of restoration ecosystem after Bowring cattleya germ plasm resource Plantlet in vitro and preservation |
| CN107278901A (en) * | 2017-07-31 | 2017-10-24 | 王生旭 | A kind of sterilization method of iris bennet bud |
| CN109601386A (en) * | 2019-01-24 | 2019-04-12 | 北京农业职业学院 | The sterilization method of bud explant is cut in a kind of state orchid stem tip tissue culture |
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