CN107750949A - The method that hybrid cymbidium seedling is efficiently bred using stem-tip tissue - Google Patents
The method that hybrid cymbidium seedling is efficiently bred using stem-tip tissue Download PDFInfo
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- CN107750949A CN107750949A CN201711018121.9A CN201711018121A CN107750949A CN 107750949 A CN107750949 A CN 107750949A CN 201711018121 A CN201711018121 A CN 201711018121A CN 107750949 A CN107750949 A CN 107750949A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- 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
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
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Abstract
The invention provides the method that hybrid cymbidium seedling is efficiently bred using stem-tip tissue, methods described includes the selection of explant；Lateral bud sterilizes；Protocorm Fiber differentiation；Protocorm Multiplication culture；Protocorm differentiation culture and Rooting and hardening-off culture.Induction period of the present invention improves protocorm inductivity；Multiplicative stage reduces protocorm aberration rate and glass rate；Differential period reduces bud aberration rate, and the development of strong plantlets and rootage stage seedling early growth is more healthy and stronger, and new root entangling ratio reduces, and root system more disperses and stretching, extension, while seedling aberration rate reduces.
Description
It is July 22, Application No. 2015104339758, entitled one in 2015 applying date that present patent application, which is,
Kind efficiently breeds the divisional application of the Chinese patent application of the method for hybrid cymbidium seedling using stem-tip tissue.
Technical field
The present invention relates to a kind of method that hybrid cymbidium seedling is efficiently bred using stem-tip tissue, more particularly to one kind can be with
Make seedling early growth development more healthy and stronger, new root entangling ratio reduction, the method for tissue culture that seedling aberration rate reduces.
Background technology
Hybrid cymbidium (Cymbidium hubridum) is that to be grown nonparasitically upon another plant kind with the big flower pattern of some in Cymbidium be parent through excessive generation
Big, bright in luster, robust growth the improved seeds group of flower pattern that cross selection cultivates.Hybrid cymbidium posture is graceful, leaf length
Dark green, flower appearance is rough, and the florescence up to 2-3 months, is epochmaking decorative indoor plant, is world-renowned " orchid nova ",
It is elegant rich and varied with cattleya to have the blue delicate fragrance of state concurrently, it is in very great demand in International Flower market, it is deep by people's all over the world
Like.
At present, protocorm and Multiple Buds are passed through mainly using the stem apex of virus-free healthy and strong maternal plant lateral bud as explant in production
Approach amount reproduction hybrid cymbidium seedling.Factorial praluction hybrid cymbidium seedling is primarily present following four problem:
First, when being bred using the stem apex of lateral bud, if stem-tip tissue is too big, sterilization is not often thorough, sterilizes successfully
Rate is extremely low, while stem-tip tissue absorption nutriment is slower, easy browning during culture.If stem-tip tissue is too small, viability is big
Big to reduce, late stage of culture is easily dead, causes protocorm inductivity relatively low.
Second, hybrid cymbidium multiplicative stage protocorm easily makes a variation, vitrification phenomenon is more serious, and this protocorm later stage is difficult
With Bud Differentiation.
Third, hybrid cymbidium differential period a part of protocorm can gradually ossify, it is mainly shown as that base portion pseudobulb can be different
It is often loose, undifferentiated leaf, or i.e. enabled differentiation leaf, but blade is smaller, it is impossible to and further growth is developed, and causes protocorm differentiation
Bud mutation it is different, produce largely rigid seedling.
Fourth, hybrid cymbidium strong plantlets and rootage stage seedling aberration rate is higher, seedling is not healthy and strong enough, can produce more inhomogeneity
Long bamboo pole seedling between the variation seedling of type, such as blade alternate or the cross seedling for arrangement of growing thickly, stipes.In addition, seedling is new
Root entangling ratio is high, is not easy to separate, and is unfavorable for later stage greenhouse transplanting domestication.
The content of the invention
The present invention is directed to disadvantage mentioned above, and large-scale technological innovation is carried out to it, there is provided one kind is efficient using stem-tip tissue
The method for breeding hybrid cymbidium seedling, optimization explant sterilization method and explant selection cutting technique, the efficient induction of design,
Propagation, differentiation, the culture medium prescription in strong plantlets and rootage stage, stem-tip tissue sterilization success rate, protocorm induction are improved so as to reach
Rate；Reduce protocorm aberration rate and glass rate；Reduce Bud Differentiation aberration rate；So that seedling early growth develops more healthy and stronger, new root
Entanglement ratio reduces, and root system is more scattered and stretches, while the purpose that seedling aberration rate reduces.
To achieve the above object, the present invention takes following technical proposals to realize：
A kind of method that hybrid cymbidium seedling is efficiently bred using stem-tip tissue, is comprised the following steps：
Step a, the selection of explant, cut the 6-12cm that hybrid cymbidium maternal plant base portion is grown healthy and strong lateral bud；
Step b, lateral bud sterilization, carries out disinfection to the lateral bud cut, cuts explant of the stem-tip tissue as culture；
Step c, protocorm Fiber differentiation, explant is inoculated into protocorm induction medium, is cultivated 30-60 days, training
It is 25-28 DEG C, light application time 8-14h/d, intensity of illumination 1000-2000lux to support temperature, and induction obtains protocorm；It is described
The composition of inducing culture includes：Spend treasured No. 3 1-3g/L, caseinhydrolysate 2-4g/L, adenine 0.5-1.5g/L, activated carbon
0.5-1.5g/L, agar 6-8g/L, sucrose 15-24g/L；
Step d, Protocorm Multiplication culture, the protocorm group of induced synthesis is cut into fritter, every piece contains 3-5 protocorm
Stem, it is inoculated into proliferated culture medium, gradually forms new protocorm, repeatedly cutting propagation, per subculture switching in 30-40 days once；
The composition of the proliferated culture medium is identical with Fiber differentiation based component；
Step e, protocorm differentiation culture, the protocorm group that the multiplicative stage is formed is cut into fritter, every piece containing 5-10
Protocorm, it is inoculated into differential medium and carries out differentiation culture, protocorm differentiation goes out young leaves new root；The differential medium into
Dividing includes：Spend treasured No. 3 1-3g/L, caseinhydrolysate 2-4g/L, adenine 0.4-1.0g/L, TDZ (Thidiazuron) 0.05-
0.15mg/L, activated carbon 0.5-1.5g/L, agar 5-9g/L, sucrose 13-26g/L.
Step f, Rooting and hardening-off culture, the bud of high 5-6cm after differentiation culture is cut, is inoculated into Rooting and hardening-off culture base
Row culture, obtains the seedling of robust growth, the composition of wherein Rooting and hardening-off culture base includes：Spend No. 1 1-4g/L of treasured, potassium nitrate
0.7-2.1g/L, ferrous sulfate heptahydrate 18-32mg/L, two water disodium ethylene diamine tetraacetate 30-43g/L, caseinhydrolysate 2-4g/
L, adenine 0.5-1.5g/L, mashed potatoes 12-28g/L, banana puree 35-65g/L, coconut milk 50-150mL, activated carbon 0.5-
1.5g/L, agar 6-8g/L, sucrose 14-27g/L.
Further, in step b, the method that lateral bud carries out disinfection is comprised the following steps：
The lateral bud cut is rinsed well with flowing water, peels off lateral bud outer blade, cutting side bastem portion lignifying group layer by layer
Knit, until remaining 6-8 piece leaves, continue to be rinsed well with flowing water, then sterilize 15min with 12% calcium hypochlorite, use blotting paper
Surface moisture is blotted, takes and is further sterilized on aseptic working platform；
Continue to peel off stem apex outer blade, until remaining 3-5 piece leaves, then sterilize 12min, nothing with 5% calcium hypochlorite
Bacterium water rinses 3-5 times；
Continue to shell outer blade, until remaining 1-2 piece leaves, then sterilize 6min, aseptic water washing 3- with 1% calcium hypochlorite
5 times, surface moisture is blotted with blotting paper；
Face under the microscope, the spire of growing point outer layer is carefully peelled off, until exposing growing point；
Cut the stem-tip tissue containing growing point, about 2-3mm sizes, the explant as culture.
Further, the stem-tip tissue containing growing point is uniformly cut into 4 fritters (cross cutting method), or 2 fritters (one from center
It is divided into two methods), every piece is individually inoculated into inducing culture.
Further, the composition of the inducing culture and proliferated culture medium includes：Spend treasured No. 3 2g/L, caseinhydrolysate 3g/
L, adenine 1g/L, activated carbon 1g/L, agar 7g/L, sucrose 20g/L.
Further, the composition of the differential medium includes：Spend treasured No. 3 2g/L, caseinhydrolysate 3g/L, adenine 1g/
L, TDZ0.1mg/L, activated carbon 1g/L, agar 7g/L, sucrose 20g/L.
Further, the composition of Rooting and hardening-off culture base includes：Spend treasured No. 1 1-4g/L, potassium nitrate 0.7-2.1g/L, seven water sulphur
Acid ferrous 18-32mg/L, two water disodium ethylene diamine tetraacetate 30-43mg/L, caseinhydrolysate 2-4g/L, adenine 0.5-
1.5g/L, mashed potatoes 12-28g/L, banana puree 35-65g/L, coconut milk 100ml/L, activated carbon 0.5-1.5g/L, agar 6-8g/
L, sucrose 14-27g/L.
Preferably, the composition of Rooting and hardening-off culture base includes：Spend treasured No. 1 2g/L, potassium nitrate 1.5g/L, ferrous sulfate heptahydrate
27.8mg/L, two water disodium ethylene diamine tetraacetate 37.3mg/L, caseinhydrolysate 3g/L, adenine 1g/L, mashed potatoes 20g/L,
Banana puree 50g/L, coconut milk 100ml/L, activated carbon 1g/L, agar 7g/L, sucrose 20g/L.
In a word, beneficial effects of the present invention are：
Cymbidium grandiforium cultural method provided by the invention, first, the multiplicative stage, the culture prepared using the present invention
Base, protocorm aberration rate, glass rate can be significantly reduced.
Differential period, the culture medium prepared using the present invention, the aberration rate of Bud Differentiation can be significantly reduced, reduces the seedling number that ossifys
Amount.
In the strong plantlets and rootage stage, the culture medium prepared using the present invention, seedling early growth can be made to develop more healthy and stronger, new root entangling
Ratio reduces, and root system is more scattered and stretches, while reduces seedling aberration rate.
Further, using the present invention sterilization method and explant selection cutting technique can improve stem-tip tissue sterilization into
Power, while increase stem-tip tissue and culture medium contact area, promote absorption of nutrient ingredients, reduce melting brown rate, improve protocorm
Inductivity.
Embodiment
With reference to specific embodiment, the present invention will be described in detail.
Embodiment 1
The composition of inducing culture and proliferated culture medium is as follows：
Spend treasured No. 3 2g/L, caseinhydrolysate 3g/L, adenine 1g/L, activated carbon 1g/L, agar 7g/L, sucrose 20g/L.
Embodiment 2
The composition of inducing culture and proliferated culture medium includes：
Spend treasured No. 3 1g/L, caseinhydrolysate 2g/L, adenine 0.5g/L, activated carbon 0.5g/L, agar 6g/L, sucrose
15g/L。
Embodiment 3
The composition of inducing culture and proliferated culture medium includes：
Spend treasured No. 3 3g/L, caseinhydrolysate 4g/L, adenine 1.5g/L, activated carbon 1.5g/L, agar 8g/L, sucrose
24g/L。
Embodiment 4
The composition of differential medium includes：
Treasured No. 3 2g/L, caseinhydrolysate 3g/L, adenine 1g/L, TDZ0.1mg/L, activated carbon 1g/L, agar 7g/L are spent,
Sucrose 20g/L.
Embodiment 5
The composition of differential medium includes：
Spend treasured No. 3 1g/L, caseinhydrolysate 2g/L, adenine 0.4g/L, TDZ0.05mg/L, activated carbon 0.5g/L, agar
5g/L, sucrose 13g/L.
Embodiment 6
The composition of differential medium includes：
Spend treasured No. 3 3g/L, caseinhydrolysate 4g/L, adenine 1.0g/L, TDZ0.15mg/L, activated carbon 1.5g/L, agar
9g/L, sucrose 26g/L.
Embodiment 7
The composition of Rooting and hardening-off culture base includes：
Spend treasured No. 1 2g/L, potassium nitrate 1.5g/L, ferrous sulfate heptahydrate 27.8mg/L, two water disodium ethylene diamine tetraacetates
37.3g/L, caseinhydrolysate 3g/L, adenine 1g/L, mashed potatoes 20g/L, banana puree 50g/L, coconut milk 100ml/L, activity
Charcoal 1g/L, agar 7g/L, sucrose 20g/L.
Embodiment 8
The composition of Rooting and hardening-off culture base includes：
Spend treasured No. 1 1g/L, potassium nitrate 0.7g/L, ferrous sulfate heptahydrate 18mg/L, two water disodium ethylene diamine tetraacetate 30mg/
L, caseinhydrolysate 2g/L, adenine 0.5g/L, mashed potatoes 12g/L, banana puree 35g/L, coconut milk 50ml/L, activated carbon
0.5g/L, agar 6g/L, sucrose 14g/L.
Embodiment 9
The composition of Rooting and hardening-off culture base includes：
Spend treasured No. 1 4g/L, potassium nitrate 2.1g/L, ferrous sulfate heptahydrate 32mg/L, two water disodium ethylene diamine tetraacetate 43mg/
L, caseinhydrolysate 4g/L, adenine 1.5g/L, mashed potatoes 28g/L, banana puree 65g/L, coconut milk 150ml/L, activated carbon
1.5g/L, agar 8g/L, sucrose 27g/L.
Embodiment 10
First, cut the healthy and strong lateral bud for the 6-12cm that hybrid cymbidium maternal plant base portion is grown.
Second, the lateral bud cut is rinsed well with flowing water, peels off lateral bud outer blade layer by layer, cutting side bastem portion is wooden
Change tissue, until being left 6 or so leaves, continue to be rinsed well with flowing water, then sterilize 15min with 12% calcium hypochlorite, use
Blotting paper blots surface moisture, takes and is further sterilized on aseptic working platform；
Continue to peel off stem apex outer blade, until being left 3 or so leaves, then sterilize 12min with 5% calcium hypochlorite,
Aseptic water washing 3-5 times；
Continue to peel off outer blade, until remaining 1-2 pieces or so leaf, then sterilize 6min, sterilized water with 1% calcium hypochlorite
Rinse 3-5 times, surface moisture is blotted with blotting paper；
Face under the microscope, the spire of growing point outer layer is carefully peelled off, until exposing growing point；
The stem-tip tissue containing growing point is cut, about 2-3mm sizes, the stem-tip tissue containing growing point is uniformly cut into from center
4 fritters (cross cutting method), every piece is individually inoculated into inducing culture as explant.
Stem-tip tissue sterilization success rate can be improved using the sterilization method of the present invention.Meanwhile explant activity is kept, no
Cause death as disinfecting time is long.
Sterilization method using the present invention can make sterilization success rate reach more than 85%, be sterilized in preferable the present embodiment
Success rate can reach more than 90%.
And the stem-tip tissue containing growing point is uniformly cut into 4 fritters from center after sterilizing, if being because explant is too big, inhale
It is slower to receive nutriment, easy browning during culture.If explant is too small, late stage of culture is easily dead, causes protocorm inductivity
It is relatively low.
Cutting technique is selected by the explant of the present invention, adds the contact area of explant and culture medium, nutrients
Matter assimilation effect is more preferable, can reduce explant browning rate, improves protocorm inductivity.3rd, protocorm Fiber differentiation, it will cut
Explant be inoculated into protocorm induction medium after, 10d or so starts to expand, and surface, which gradually induces, produces many light green colors
Point-like particle, 25d or so develop into green protocorm.Protocorm full grains, surface are covered with white fluff.The induction training
Support the culture medium in base selection example 1.Cultivation cycle is 30-60d, and cultivation temperature is 25-28 DEG C, light application time 8-14h/
D, intensity of illumination 1000-2000lux.The Fiber differentiation based component and cultural method of the present invention is sampled, protocorm inductivity is big
In 95%.
4th, Protocorm Multiplication culture, the protocorm group of induced synthesis is cut into fritter, every piece contains 3-5 protocorm,
It is inoculated into proliferated culture medium, gradually forms new protocorm, repeatedly cutting propagation；Transfer 1 time, formed big per 30-40d subcultures
The protocorm of amount；The Multiplying culture based component is identical with Fiber differentiation based component, the culture medium in same selection example 1.
Condition of culture is identical with the Fiber differentiation stage.
Multiplicative stage normotrophic protocorm is light green, rough and have tiny projection, and short white is covered on surface
Fine hair, protocorm later stage easy Bud Differentiation.
And the protocorm to make a variation is bottle green or blackish green, surface is smooth and glossy.Meanwhile glass occurs for some protocorms
Glass phenomenon, transparent shape.The protocorm surface of variation is without white fluff, although the speed of growth is quickly, vigor is very low, the later stage
Bud Differentiation ratio is low, or even is unable to Bud Differentiation.
And the Multiplying culture based component of the present invention and cultural method is used to significantly reduce protocorm aberration rate, vitrifying
Rate.Growth coefficient is between 4-5, and aberration rate is less than 3%, and glass rate is less than 1%, and aberration rate is less than in preferable the present embodiment
2%.
5th, protocorm differentiation culture, the protocorm group that the multiplicative stage is formed is cut into fritter, every piece is former containing 5-10
Bulb, it is inoculated into differential medium and carries out differentiation culture, protocorm differentiation goes out young leaves new root；The differential medium selection is real
Apply the culture medium of example 4.Cultivation cycle is 40-60d, and cultivation temperature is 25-28 DEG C, light application time 8-14h/d, and intensity of illumination is
2000-3000lux。
Differential period, a part of protocorm can gradually ossify, show as base portion pseudobulb can aberrant mast, undifferentiated leaf, or
The i.e. enabled differentiation leaf of person, but blade is smaller, it is impossible to and further growth is developed, and causes the bud mutation of protocorm differentiation different, is produced a large amount of
Rigid seedling.This seedling leaf is harder, surface as dehydration, the later stage normally can not take root and develop.
Rigid seedling ratio can greatly be reduced using the differential medium composition and cultural method of the present invention, ossify seedling ratio
Between 2%-4%, it is preferred that less than 2%.
6th, the bud of plant height 5-6cm after differentiation culture is chosen, is inoculated into Rooting and hardening-off culture base and is cultivated, wherein
Rooting and hardening-off culture base selection example 7.Cultivation cycle is 60-80d, and cultivation temperature is 25-28 DEG C, light application time 8-14h/
D, intensity of illumination 2500-3000lux.
The hybrid cymbidium strong plantlets and rootage stage, seedling aberration rate was higher, and seedling is not healthy and strong enough, and part kind seedling leaf is loose not
It is straight and upright, fragile, plant is more slim and frahile.In addition, the strong plantlets and rootage stage can also produce two kinds of variation seedling, such as blade alternate
Or long bamboo pole seedling between the cross seedling of arrangement of growing thickly, stipes.Rooting and hardening-off culture based component and culture using the present invention
Method can greatly reduce the ratio of the variating seedlings such as cross seedling, bamboo pole seedling, and variating seedling rate is less than 3%.Moreover, kind seedling leaf is straight
Stand, stem is sturdy, plant type is tall and straight.Average plant height is 8-10cm, and the number of blade is more than 4, and radical is more than 4.
In addition, strong plantlets and rootage stage preferable seedling, root system is more dispersed, root system stretching, extension is good, and bottle outlet is easy to clean, no
Easily cause damage.
And the seedling that developmental condition is bad, root system are entangled, it is not easy to separate, some root system tips can expand balling
Shape, it is unfavorable for later stage greenhouse transplanting domestication.
Root system entanglement ratio can be significantly reduced using the Rooting and hardening-off culture based component and cultural method of the present invention, tangled
Ratio is less than 3%.
Although the present invention is disclosed as above with preferred embodiment, it is not for limiting the present invention, any this area
Technical staff without departing from the spirit and scope of the present invention, may be by the methods and technical content of the disclosure above to this hair
Bright technical scheme makes possible variation and modification, therefore, every content without departing from technical solution of the present invention, according to the present invention
Technical spirit to any simple modifications, equivalents, and modifications made for any of the above embodiments, belong to technical solution of the present invention
Protection domain.
Claims (5)
- A kind of 1. method that hybrid cymbidium seedling is efficiently bred using stem-tip tissue, it is characterised in that methods described includes following Step：Step a, the selection of explant, cut the 6-12cm that hybrid cymbidium maternal plant base portion is grown healthy and strong lateral bud；Step b, lateral bud sterilization, carries out disinfection to the lateral bud cut, cuts explant of the stem-tip tissue as culture；Step c, protocorm Fiber differentiation, explant is inoculated into protocorm induction medium, is cultivated 30-60 days, culture temperature Spend for 25-28 DEG C, light application time 8-14h/d, intensity of illumination is that 1000-2000lux induces to obtain protocorm；The induction training Supporting the composition of base includes：The inducing culture and proliferated culture medium consist of the following composition：No. 3 2g/L of treasured are spent, hydrolyze junket egg White 3g/L, adenine 1g/L, activated carbon 1g/L, agar 7g/L, sucrose 20g/L；Step d, Protocorm Multiplication culture, the protocorm group of induced synthesis is cut into fritter, every piece contains 3-5 protocorm, connects Kind gradually forms new protocorm into proliferated culture medium, repeatedly cutting propagation, per subculture switching in 30-40 days once；The increasing The composition for growing culture medium is identical with Fiber differentiation based component；Step e, protocorm differentiation culture, the protocorm group that propagation obtains is cut into fritter, every piece contains 5-10 protocorm, connects Kind carries out differentiation culture into differential medium, and protocorm differentiation goes out young leaves new root；Differential medium consists of the following composition：Flower Treasured No. 3 2g/L, caseinhydrolysate 3g/L, adenine 1g/L, TDZ0.1mg/L, activated carbon 1g/L, agar 7g/L, sucrose 20g/L；Step f, Rooting and hardening-off culture, the bud of high 5-6cm after differentiation culture is cut, is inoculated into Rooting and hardening-off culture base and is trained Support, obtain the seedling of robust growth, wherein Rooting and hardening-off culture base consists of the following composition：Spend No. 1 1-4g/L of treasured, potassium nitrate 0.7-2.1g/L, ferrous sulfate heptahydrate 18-32mg/L, two water disodium ethylene diamine tetraacetate 30-43g/L, caseinhydrolysate 2-4g/ L, adenine 0.5-1.5g/L, mashed potatoes 12-28g/L, banana puree 35-65g/L, coconut milk 50-150mL, activated carbon 0.5- 1.5g/L, agar 6-8g/L, sucrose 14-27g/L.
- 2. according to the method for claim 1, it is characterised in that in step b, the method to be carried out disinfection to lateral bud is including following Step：The lateral bud cut is rinsed well with flowing water, peels off outer blade, cutting side bastem portion lignified tissue, until surplus layer by layer Lower 6-8 piece leaves, 12% calcium hypochlorite sterilization 15mi n, then take on aseptic working platform and further sterilize；Continue to peel off stem apex outer blade, until remaining 3-5 piece leaves, 5% calcium hypochlorite sterilization 12mi n；Continue to shell outer blade, until remaining 1-2 piece leaves, 1% calcium hypochlorite sterilization 6mi n；Face under the microscope, the spire of growing point outer layer is carefully peelled off, until exposing growing point；The stem-tip tissue containing growing point is cut, the explant as culture.
- 3. according to the method for claim 2, it is characterised in that the stem-tip tissue containing growing point is uniformly cut into 4 from center Fritter, or 2 fritters, every piece is individually inoculated into inducing culture.
- 4. according to the method for claim 1, it is characterised in that the composition of Rooting and hardening-off culture base includes：Spend treasured No. 1 1-4g/L, potassium nitrate 0.7-2.1g/L, ferrous sulfate heptahydrate 18-32mg/L, two water disodium ethylene diamine tetraacetates 30-43g/L, caseinhydrolysate 2-4g/L, adenine 0.5-1.5g/L, mashed potatoes 12-28g/L, banana puree 35-65g/L, coconut Juice 100ml/L, activated carbon 0.5-1.5g/L, agar 6-8g/L, sucrose 14-27g/L.
- 5. according to the method for claim 5, it is characterised in that the composition of Rooting and hardening-off culture base includes：Spend No. 1 2g/ of treasured L, potassium nitrate 1.5g/L, ferrous sulfate heptahydrate 27.8mg/L, two water disodium ethylene diamine tetraacetate 37.3g/L, caseinhydrolysate 3g/ L, adenine 1g/L, mashed potatoes 20g/L, banana puree 50g/L, coconut milk 100ml/L, activated carbon 1g/L, agar 7g/L, sucrose 20g/L。
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109042342A (en) * | 2018-11-01 | 2018-12-21 | 翁源县天下泽雨农业科技有限公司 | A kind of method for tissue culture of Chunlan |
| 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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| CN113475396A (en) * | 2021-07-25 | 2021-10-08 | 杭州市农业科学研究院 | Efficient seedling method for hybrid cymbidium hybridum of spring sword and cymbidium hybridum |
| CN114532225A (en) * | 2022-02-28 | 2022-05-27 | 广西壮族自治区农业科学院 | Rapid propagation and cultivation method for paphiopedilum delavayi tissue culture |
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| CN109804927B (en) * | 2019-03-05 | 2020-06-05 | 湖北红杏生态环境科技有限公司 | In-vitro rapid propagation method of cymbidium hybridum |
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| CN113475396A (en) * | 2021-07-25 | 2021-10-08 | 杭州市农业科学研究院 | Efficient seedling method for hybrid cymbidium hybridum of spring sword and cymbidium hybridum |
| CN114532225A (en) * | 2022-02-28 | 2022-05-27 | 广西壮族自治区农业科学院 | Rapid propagation and cultivation method for paphiopedilum delavayi tissue culture |
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| CN104938341A (en) | 2015-09-30 |
| CN107750949B (en) | 2019-10-11 |
| CN104938341B (en) | 2017-12-12 |
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