CN110178733A - A kind of the quick reproduction technique culture medium and quick reproduction technique method of mountain orchid protocorm - Google Patents
A kind of the quick reproduction technique culture medium and quick reproduction technique method of mountain orchid protocorm Download PDFInfo
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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
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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
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Abstract
The present invention relates to technical field of tissue culture, in particular to the quick reproduction technique culture medium and quick reproduction technique method of a kind of mountain orchid protocorm.The quick reproduction technique culture medium includes, and extends proliferated culture medium: containing sucrose, agar powder, NAA, ZT MS culture medium；Differential medium: containing sucrose, agar powder, IAA, 6-BA MS culture medium；Root media: containing sucrose, agar powder, IAA, 6-BA MS culture medium.Under condition of tissue culture of the present invention, by hormone regulating and controlling, the protocorm speed of growth is fast, and branched elongation growth coefficient reaches 8.67, protocorm differentiation generates bulb coefficient and reaches 5.14, i.e. 1 protocorm can produce about 45 root, stem and leafs completely healthy and strong seedling after shoot proliferation and differentiation culture.
Description
Technical field
The present invention relates to technical field of tissue culture, in particular to the quick reproduction technique culture medium and kind of a kind of mountain orchid protocorm
Seedling fast reproducing method.
Background technique
Mountain orchid Oreorchis patens (Lindl.) Lindl. is the perennial raw herbaceous plant of yin of orchid family mountain Cymbidium, extensively
Southwestern China, Central China, northwest and northeast various regions are distributed in, the ground such as the Korea peninsula to Siberia, Japan also have a small amount of point
Cloth.Its drying bulb is used as medicine, sweet, pungent, trembles with fear, slightly poisonous.Have effects that enriching yin clearing lung-heat, preventing phlegm from forming and stopping coughing, is used for carbuncle infantile malnutrition due to digestive disturbances or intestinalparasites sore swells, scrofula
Scrofula, nameless sores or boils, but the medical value of mountain orchid is not taken seriously for a long time, but be taken as edible tulip adulterant treat or
It is used as medicine as the product of commonly using.As mountain orchid demand increased in recent years, big bulge in price causes its going to wreck property of wild resource broken
It is bad, it is listed in Jilin Province three-level protective plant, about its development of resources, effective component, pharmacological activity, wild domestication and mycorhiza
The correlative study of fungi etc. is more, but the seedling breeding key technology research report for restricting mountain orchid large area artificial cultivation is few.
Tissue culture technique be under the aseptic condition artificially created by the isolated organ of life (such as root, stem, leaf, stem section,
Protoplast), tissue or cell be placed in culture medium, and be placed in suitable environment, carry out continuous culture to obtain cell, group
Knit or individual technology.This technology is widely used to the research of agricultural and biology, medicine.
In the orchid tissue culture technique of mountain, there is the Multiplying culture for carrying out protocorm as explant using pseudobulb or blade
Public technology, as the Chinese patent of Publication No. CN107736245A discloses a kind of mountain orchid Fast Asexual Propagation Technique, the technology
Make explant mass production mountain orchid test tube seedling with mountain orchid pseudobulb or blade or peduncle.It is proliferated, is proliferated in a manner of bulb budding
The coefficient low period is long, and it is high that proliferation growth coefficient is carried out in the form of protocorm, but has been proliferated mostly an original with bulb budding mode
Bulb is divided into the step of bulb.
Summary of the invention
In view of this, the present invention provides a kind of quick reproduction technique culture medium of mountain orchid protocorm and quick reproduction technique methods.It should
The protocorm that invention was generated using 1 year green-ball stem is studied 4 kinds of auxin and mitogen respectively and is broken up to Protocorm Multiplication as explant
Influence and hormone combinations elongation Proliferation, Differentiation branched to protocorm, protocorm differentiation generate the shadow that bulb and bulb are taken root
It rings, forms a complete mountain orchid protocorm approach quick reproduction technique technical system.
In order to achieve the above-mentioned object of the invention, the present invention the following technical schemes are provided:
The present invention also provides a kind of quick reproduction technique culture mediums of mountain orchid protocorm, comprising:
Extend proliferated culture medium: containing 28~32g/L of sucrose, 4.0~5.0g/L of agar powder, 0.5~1.5mg/L of NAA,
The MS culture medium of 0.5~1.5mg/L of ZT；
Differential medium: contain 28~32g/L of sucrose, 4.0~5.0g/L of agar powder, 2.0~3.0mg/L of IAA, 6-BA
The MS culture medium of 2.0~3.0mg/L；
Root media: contain 28~32g/L of sucrose, 4.0~5.0g/L of agar powder, 2.0~3.0mg/L of IAA, 6-BA
The MS culture medium of 0.05~0.15mg/L.
Preferably, the quick reproduction technique culture medium includes:
Extend proliferated culture medium: the MS containing sucrose 30g/L, agar powder 4.5g/L, NAA 1.0mg/L, ZT 1.0mg/L
Culture medium；
Differential medium: the MS training containing sucrose 30g/L, agar powder 4.5g/L, IAA 2.5mg/L, 6-BA 2.5mg/L
Support base；
Root media: the MS training containing sucrose 30g/L, agar powder 4.5g/L, IAA 2.5mg/L, 6-BA 0.1mg/L
Support base.
The present invention also provides a kind of quick reproduction technique methods of mountain orchid protocorm, comprising:
Elongation Multiplying culture is carried out to protocorm explant using elongation proliferated culture medium, the protocorm after obtaining elongation proliferation
Stem；Elongation proliferated culture medium is to contain 28~32g/L of sucrose, 4.0~5.0g/L of agar powder, 0.5~1.5mg/L of NAA, ZT
The MS culture medium of 0.5~1.5mg/L；
Differentiation culture is carried out to the protocorm after elongation proliferation using differential medium, obtains bulb；Differential medium is
MS training containing 28~32g/L of sucrose, 4.0~5.0g/L of agar powder, 2.0~3.0mg/L of IAA, 2.0~3.0mg/L of 6-BA
Support base；
Culture of rootage is carried out to bulb using root media；Root media is to contain 28~32g/L of sucrose, agar powder
The MS culture medium of 4.0~5.0g/L, 2.0~3.0mg/L of IAA, 0.05~0.15mg/L of 6-BA.
Preferably, elongation proliferated culture medium is to contain sucrose 30g/L, agar powder 4.5g/L, NAA 1.0mg/L, ZT
The MS culture medium of 1.0mg/L；
Differential medium is the MS training containing sucrose 30g/L, agar powder 4.5g/L, IAA 2.5mg/L, 6-BA 2.5mg/L
Support base；
Root media is the MS training containing sucrose 30g/L, agar powder 4.5g/L, IAA 2.5mg/L, 6-BA 0.1mg/L
Support base.
Preferably, the size of explant is (2~4) × (2~4) × (4~6) mm.
Preferably, the size of explant is 3 × 3 × 5mm.
Preferably, protocorm is the protocorm that 1 year green-ball stem generates.
Preferably, the condition of elongation Multiplying culture are as follows: cultivation temperature is 22~24 DEG C, and light dark period is 13~15h/9
~11h, intensity of illumination are 1400~1600lx, 50~60d of incubation time.
Preferably, the condition of Multiplying culture is extended are as follows: cultivation temperature is 23 DEG C, light dark period 14h/10h, intensity of illumination
For 1500lx, incubation time 55d.
Preferably, differentiation culture condition are as follows: cultivation temperature be 22~24 DEG C, light dark period be 13~15h/9~
11h, intensity of illumination are 1400~1600lx, 50~60d of incubation time.
Preferably, break up the condition of culture are as follows: cultivation temperature is 23 DEG C, light dark period 14h/10h, and intensity of illumination is
1500lx, incubation time 55d.
Preferably, the condition of culture of rootage are as follows: cultivation temperature be 22~24 DEG C, light dark period be 13~15h/9~
11h, intensity of illumination are 1400~1600lx, 50~60d of incubation time.
Preferably, the condition of culture of rootage are as follows: cultivation temperature is 23 DEG C, light dark period 14h/10h, and intensity of illumination is
1500lx, incubation time 55d.
The present invention provides a kind of quick reproduction technique culture medium of mountain orchid protocorm and quick reproduction technique methods.Quick reproduction technique training
Feeding base includes, and extends proliferated culture medium: containing 28~32g/L of sucrose, 4.0~5.0g/L of agar powder, 0.5~1.5mg/L of NAA,
The MS culture medium of 0.5~1.5mg/L of ZT；Differential medium: contain 28~32g/L of sucrose, 4.0~5.0g/L of agar powder, IAA
The MS culture medium of 2.0~3.0mg/L, 2.0~3.0mg/L of 6-BA；Root media: contain 28~32g/L of sucrose, agar powder
The MS culture medium of 4.0~5.0g/L, 2.0~3.0mg/L of IAA, 0.05~0.15mg/L of 6-BA.The technology effect that the present invention has
Fruit are as follows:
The protocorm that the application test was generated using 1 year green-ball stem, using MS as minimal medium, studies 4 as explant respectively
Influence that kind of auxin and mitogen break up Protocorm Multiplication and hormone combinations elongation Proliferation, Differentiation branched to protocorm,
Protocorm differentiation generates the influence that bulb and bulb are taken root.The result shows that NAA and ZT is conducive to the branched elongation proliferation of protocorm,
The branched elongation growth coefficient of protocorm reaches 8.67 when NAA 1.0mg/L+ZT1.0mg/L；IAA and 6-BA is conducive to protocorm point
Bulb is turned to, protocorm differentiation is that bulb coefficient reaches 5.14 in IAA 2.5mg/L+6-BA2.5mg/L；IAA and 6-BA benefit
It takes root in the bulb that protocorm differentiation generates, in IAA 2.5mg/L+6-BA 0.1mg/L, bulb generation radical reaches 6.26
Item/bulb, root weigh to 9.90mg/ item.
The protocorm that this test was generated using 1 year green-ball stem studies 4 kinds of auxin and mitogen to protocorm as explant respectively
The influence of stem Proliferation, Differentiation and hormone combinations elongation Proliferation, Differentiation branched to protocorm, protocorm differentiation generate bulb and ball
The influence of stem-root forms a complete mountain orchid protocorm approach quick reproduction technique technical system.
Under natural conditions, the branched proliferation slow growth of mountain orchid protocorm and 1 bulb, natural propagation efficiency are only differentiated
It is low；Under condition of tissue culture, by hormone regulating and controlling, the protocorm speed of growth is fast, and branched elongation growth coefficient reaches 8.67, protocorm
Differentiation generates bulb coefficient and reaches 5.14, i.e. 1 protocorm can produce about 45 rhizomes after shoot proliferation and differentiation culture
The complete healthy and strong seedling of leaf.
Detailed description of the invention
Fig. 1 shows the branched elongation proliferation of protocorm；
Fig. 2 shows that protocorm differentiation generates bulb；
Fig. 3 shows that bulb differentiation generates root.
Specific embodiment
The invention discloses a kind of quick reproduction technique culture medium of mountain orchid protocorm and quick reproduction technique method, those skilled in the art
Member can use for reference present disclosure, be suitably modified realization of process parameters.In particular, it should be pointed out that all similar substitutions and modifications
Apparent to those skilled in the art, they are considered as being included in the present invention.Method and application of the invention
Be described by preferred embodiment, related personnel obviously can not depart from the content of present invention, in spirit and scope it is right
Method described herein and application are modified or appropriate changes and combinations, carry out implementation and application the technology of the present invention.
Seed used, training in the quick reproduction technique culture medium and quick reproduction technique method of a kind of mountain orchid protocorm provided by the invention
Support base component, reagent is available on the market.
Below with reference to embodiment, the present invention is further explained:
Embodiment 1
1 test material and method
1.1 test material
For the protocorm generated using 1 year green-ball stem as explant, bulb disinfection and cultural method refer to pertinent literature (Wang Ping
Flat, Wang Yujiao, Chen Xuhui wait the north research [J] the gardening of the mountain orchid mycorrhizal fungi isolated culture condition, and 2012, (09): 66-
69.), explant is the protocorm with growing point, size about 3*3*5mm.
The influence that 1.2 auxin break up Protocorm Multiplication
Using MS+6-BA0.1mg/L+ sucrose 30g/L+ agar powder 4.5g/L as minimal medium, add respectively 2,4-D,
NAA, IBA, IAA each 0.1mg/L, 2.0mg/L, 4.0mg/L, 6.0mg/L, 8.0mg/L, 10.0mg/L；Cultivation temperature is 23 DEG C,
Light dark period is 14h/10h, intensity of illumination 1500lx, incubation time 55d, and every processing 3 repeats, 15 explants of every repetition,
Statistical indicator is the branched elongation Proliferation, Differentiation coefficient of protocorm, protocorm differentiation generates bulb coefficient and protocorm differentiation generates
Bulb is averagely taken root number, the branched elongation branch amount/inoculation protocorm of the branched elongation Proliferation, Differentiation coefficient=protocorm of protocorm
Stem number, protocorm differentiation generate bulb number that the branched elongation differentiation of bulb coefficient=protocorm generates/protocorm is branched stretches
Long branch amount, protocorm differentiation generate bulb be averaged take root number=protocorm differentiation generate bulb take root number/protocorm divide
Change the bulb number generated.
The influence that 1.3 mitogens break up Protocorm Multiplication
Using MS+NAA0.1mg/L+ sucrose 30g/L+ agar powder 4.5g/L as minimal medium, TDZ, ZT, 6- are added respectively
BA, KT each 0.1mg/L, 2.0mg/L, 4.0mg/L, 6.0mg/L, 8.0mg/L, 10.0mg/L；Cultivation temperature is 23 DEG C, brightness week
Phase is 14h/10h, intensity of illumination 1500lx, incubation time 55d, and every processing 3 repeats, and 15 explants of every repetition, statistics refers to
Mark is the same as 1.2.
The influence of 1.4 NAA and ZT combination elongation Proliferation, Differentiation branched to protocorm
Using MS+ sucrose 30g/L+ agar powder 4.5g/L as minimal medium, to the branched elongation Proliferation, Differentiation of protocorm
Effect good NAA and ZT is combined, and NAA concentration is 0.1mg/L, 1.0mg/L, 2.0mg/L, 3.0mg/L, 4.0mg/L, and ZT is dense
Degree is 0.1mg/L, 1.0mg/L, 2.0mg/L, 3.0mg/L, 4.0mg/L；Cultivation temperature is 23 DEG C, light dark period 14h/10h,
Intensity of illumination is 1500lx, and incubation time 55d, every 3 repetition of processing, 15 explants of every repetition, statistical indicator is protocorm point
Dendritic elongation Proliferation, Differentiation coefficient.
1.5 IAA and 6-BA combination generates the influence that bulb and bulb are taken root to protocorm differentiation
Using MS+ sucrose 30g/L+ agar powder 4.5g/L as minimal medium, to generate bulb and bulb to protocorm differentiation
Rooting efficiency good IAA and 6-BA is combined, IAA concentration be 0.1mg/L, 2.5mg/L, 5.0mg/L, 7.5mg/L,
10.0mg/L, 6-BA concentration are 0.1mg/L, 2.5mg/L, 5.0mg/L, 7.5mg/L, 10.0mg/L；Cultivation temperature is 23 DEG C, light
The dark period is 14h/10h, intensity of illumination 1500lx, incubation time 55d, and every processing 3 repeats, 15 explants of every repetition, system
Counting index is the several and equal root weight of taking root that is averaged that protocorm differentiation generates bulb coefficient and protocorm differentiation generates bulb, equal root weight
The total root weight/total radical of=bulb.
The analysis of 1.6 data
Test data carries out variance analysis with SAS8.01.
2 results and analysis
The influence that 2.1 auxin break up Protocorm Multiplication
2.1.1 the influence of auxin elongation Proliferation, Differentiation branched to protocorm is as shown in table 1, branched to protocorm to stretch
The significant auxin of long cultivation effect is NAA, and it is 4.15 that maximum value proliferation is reached in NAA2.0mg/L.
The influence of 1 auxin of table elongation Proliferation, Differentiation branched to protocorm
2.1.2 the influence that auxin generates bulb to protocorm differentiation is as shown in table 2, generates bulb effect to protocorm differentiation
The preferable auxin of fruit is IAA, and maximum coefficient of differentiation 0.75 is reached in IAA 6.0mg/L.
2 auxin of table generates the influence of bulb to protocorm differentiation
2.1.3 the influence that auxin takes root to protocorm differentiation generation bulb is as shown in table 3, generates ball to protocorm differentiation
The preferable auxin of stem-root effect is IAA, and maximum is reached in IAA6.0mg/L and is taken root 1.67/bulb of number.
3 auxin of table generates the influence that bulb is taken root to protocorm differentiation
The influence that 2.2 mitogens break up Protocorm Multiplication
2.2.1 the influence of mitogen elongation Proliferation, Differentiation branched to protocorm is as shown in table 4, and it is branched to be conducive to protocorm
The mitogen of elongation proliferation is ZT, and maximum growth coefficient 2.08 is reached in ZT 2.0mg/L.
The influence of 4 mitogen of table elongation Proliferation, Differentiation branched to protocorm
2.2.2 the influence that mitogen generates bulb to protocorm differentiation is as shown in table 5, generates bulb effect to protocorm differentiation
The significant mitogen of fruit is 6-BA, and maximum coefficient of differentiation 0.56 is reached in 6-BA6.0mg/L.
5 mitogen of table generates the influence of bulb to protocorm differentiation
2.2.3 the influence that mitogen takes root to protocorm differentiation generation bulb is as shown in table 6, generates ball to protocorm differentiation
The preferable mitogen of stem-root effect is 6-BA, and 1.81/bulb of maximum value is reached in 6-BA6.0mg/L.
6 mitogen of table generates the influence that bulb is taken root to protocorm differentiation
The influence of 2.3 NAA and ZT combination elongation Proliferation, Differentiation branched to protocorm
The influence of NAA and ZT combination elongation Proliferation, Differentiation branched to protocorm is as shown in table 7, former when NAA is 1.0mg/L
The branched elongation Proliferation, Differentiation coefficient of bulb is in first to increase the trend subtracted afterwards with the increase of ZT concentration, in NAA 1.0mg/L+ZT1.0mg/
The branched elongation cultivation effect of protocorm is preferable when L, as shown in Figure 1, growth coefficient reaches 8.67.
The influence of table 7 NAA and ZT combination elongation Proliferation, Differentiation branched to protocorm
2.4 IAA and 6-BA combination generates bulb to protocorm differentiation and bulb is taken root and the influence of root weight
2.4.1 the influence that IAA and 6-BA combination generates bulb to protocorm differentiation is as shown in table 8, when IAA is 2.5mg/L
The coefficient that protocorm differentiation generates bulb increases with 6-BA concentration in the trend subtracted afterwards is first increased, in IAA 2.5mg/L+6-
Protocorm differentiation is that bulb effect is preferable when BA2.5mg/L, and coefficient of differentiation reaches 5.14, as shown in Figure 2.
Table 8 IAA and 6-BA combination generate the influence of bulb to protocorm differentiation
2.4.2 the influence that IAA and 6-BA combination takes root to protocorm differentiation generation bulb is as shown in table 9, and IAA is
The bulb for being conducive to protocorm differentiation generation when 2.5mg/L is taken root, and number of taking root increases with 6-BA concentration and reduced, in IAA
Bulb takes root number at most when 2.5mg/L+6-BA 0.1mg/L, reaches 6.26/bulb, as shown in Figure 3.
Table 9 IAA and 6-BA combination generate the influence (item/bulb) that bulb is taken root to protocorm differentiation
2.4.3 the influence that IAA and 6-BA combination generates bulb root weight to protocorm differentiation is as shown in table 10, and IAA is
Be conducive to root growth when 2.5mg/L, root weight increases with 6-BA concentration and reduced, the ball in IAA2.5mg/L+6-BA0.1mg/L
The root weight that stem generates is maximum, reaches 9.90mg/ item.
The influence (mg/ item) for the bulb root weight that table 10 IAA and 6-BA combination generate protocorm differentiation
3 discuss
3.1 mountain orchid Protocorm Multiplication coefficients are high, are a kind of more satisfactory fertile materials, but directly obtained from soil
Protocorm has that disinfection difficulty is low with survival rate as inoculation material primary, and 1 year green-ball stem surface is smooth, is easy to disappear
Malicious and vitality is vigorous, therefore is that inoculation material primary is more appropriate with 1 year green-ball stem, is differentiated with 1 year green-ball stem sterile
Protocorm is as shoot proliferation material.
3.2 under condition of tissue culture, and by hormone regulating and controlling, protocorm is proliferated with callus shape and branched elongation two ways, point
The operation easy to produce of dendritic elongation Reproduction methods is the main Reproduction methods of protocorm approach quick reproduction technique.
3.3 in single factor experiment, is bulb and the best hormone concentration of bulb rooting efficiency to protocorm differentiation, in group
Best differentiation effect is not shown under the conditions of conjunction, but differentiation effect is best in lower concentration combination, this may be high
Hormone concentration the reason of there are the effects of antagonism.
3.4 grow since root crimps, and length is not easy to measure, but the diameter of root is roughly the same, therefore with the quantity and root of root
Counterpoise two indices measure rooting efficiency.
Under 3.5 natural conditions, the branched proliferation slow growth of mountain orchid protocorm and 1 bulb, natural propagation are only differentiated
Low efficiency；Under condition of tissue culture, by hormone regulating and controlling, the protocorm speed of growth is fast, and branched elongation growth coefficient reaches 8.67, original
Bulb differentiation generates bulb coefficient and reaches 5.14, i.e. 1 protocorm can produce about 45 after shoot proliferation and differentiation culture
The complete healthy and strong seedling of root, stem and leaf.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (10)
1. a kind of quick reproduction technique culture medium of mountain orchid protocorm characterized by comprising
Elongation proliferated culture medium: contain 28~32g/L of sucrose, 4.0~5.0g/L of agar powder, 0.5~1.5mg/L of NAA, ZT
The MS culture medium of 0.5~1.5mg/L；
Differential medium: contain 28~32g/L of sucrose, 4.0~5.0g/L of agar powder, 2.0~3.0mg/L of IAA, 6-BA 2.0
The MS culture medium of~3.0mg/L；
Root media: contain 28~32g/L of sucrose, 4.0~5.0g/L of agar powder, 2.0~3.0mg/L of IAA, 6-BA 0.05
The MS culture medium of~0.15mg/L.
2. quick reproduction technique culture medium according to claim 1 characterized by comprising
Extend proliferated culture medium: the MS culture containing sucrose 30g/L, agar powder 4.5g/L, NAA 1.0mg/L, ZT 1.0mg/L
Base；
Differential medium: the MS culture medium containing sucrose 30g/L, agar powder 4.5g/L, IAA 2.5mg/L, 6-BA 2.5mg/L；
Root media: the MS culture medium containing sucrose 30g/L, agar powder 4.5g/L, IAA 2.5mg/L, 6-BA 0.1mg/L.
3. a kind of quick reproduction technique method of mountain orchid protocorm characterized by comprising
Elongation Multiplying culture is carried out to protocorm explant using elongation proliferated culture medium, the protocorm after obtaining elongation proliferation；
The elongation proliferated culture medium is to contain 28~32g/L of sucrose, 4.0~5.0g/L of agar powder, 0.5~1.5mg/L of NAA, ZT
The MS culture medium of 0.5~1.5mg/L；
Differentiation culture is carried out to the protocorm after elongation proliferation using differential medium, obtains bulb；The differential medium is
MS training containing 28~32g/L of sucrose, 4.0~5.0g/L of agar powder, 2.0~3.0mg/L of IAA, 2.0~3.0mg/L of 6-BA
Support base；
Culture of rootage is carried out to bulb using root media；The root media is to contain 28~32g/L of sucrose, agar powder
The MS culture medium of 4.0~5.0g/L, 2.0~3.0mg/L of IAA, 0.05~0.15mg/L of 6-BA.
4. quick reproduction technique method according to claim 3, which is characterized in that the elongation proliferated culture medium is to contain sucrose
The MS culture medium of 30g/L, agar powder 4.5g/L, NAA 1.0mg/L, ZT 1.0mg/L；
The differential medium is the MS training containing sucrose 30g/L, agar powder 4.5g/L, IAA 2.5mg/L, 6-BA 2.5mg/L
Support base；
The root media is the MS training containing sucrose 30g/L, agar powder 4.5g/L, IAA 2.5mg/L, 6-BA 0.1mg/L
Support base.
5. quick reproduction technique method according to claim 3, which is characterized in that the size of the explant is (2~4) × (2
~4) × (4~6) mm.
6. quick reproduction technique method according to claim 3, which is characterized in that the protocorm is that 1 year green-ball stem generates
Protocorm.
7. according to the method described in claim 3, it is characterized in that, the condition of the elongation Multiplying culture are as follows: cultivation temperature is
22~24 DEG C, light dark period is 13~15h/9~11h, and intensity of illumination is 1400~1600lx, 50~60d of incubation time.
8. the method according to any one of claim 3 to 7, which is characterized in that the condition of the elongation Multiplying culture are as follows:
Cultivation temperature is 23 DEG C, light dark period 14h/10h, intensity of illumination 1500lx, incubation time 55d.
9. according to the method described in claim 3, it is characterized in that, it is described differentiation culture condition are as follows: cultivation temperature be 22~
24 DEG C, light dark period is 13~15h/9~11h, and intensity of illumination is 1400~1600lx, 50~60d of incubation time.
10. according to the method described in claim 3, it is characterized in that, the condition of the culture of rootage are as follows: cultivation temperature be 22~
24 DEG C, light dark period is 13~15h/9~11h, and intensity of illumination is 1400~1600lx, 50~60d of incubation time.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112753575A (en) * | 2020-12-31 | 2021-05-07 | 湖北金水源农业开发有限公司 | High-yield Cremastra appendiculata seedling cultivation method |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1817110A (en) * | 2006-01-18 | 2006-08-16 | 贵州省农业科学院生物技术研究所 | Cyrtopterin tissue culturing method and fast reproduction thereof |
| CN105941145A (en) * | 2016-05-04 | 2016-09-21 | 中国农业科学院特产研究所 | Germination method of Oreorchis patens seeds |
| CN109122325A (en) * | 2018-11-09 | 2019-01-04 | 翁源县天下泽雨农业科技有限公司 | A kind of aseptic seeding quick-breeding method of sword-leaved cymbidium seed |
-
2019
- 2019-07-16 CN CN201910641383.3A patent/CN110178733B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1817110A (en) * | 2006-01-18 | 2006-08-16 | 贵州省农业科学院生物技术研究所 | Cyrtopterin tissue culturing method and fast reproduction thereof |
| CN105941145A (en) * | 2016-05-04 | 2016-09-21 | 中国农业科学院特产研究所 | Germination method of Oreorchis patens seeds |
| CN109122325A (en) * | 2018-11-09 | 2019-01-04 | 翁源县天下泽雨农业科技有限公司 | A kind of aseptic seeding quick-breeding method of sword-leaved cymbidium seed |
Non-Patent Citations (2)
| Title |
|---|
| 仲磊等: "铁皮石斛兰的袋式组织培养", 《林业科技开发》 * |
| 邱玥等: "金线兰种子萌发及其原球茎的增殖培养", 《广西植物》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112753575A (en) * | 2020-12-31 | 2021-05-07 | 湖北金水源农业开发有限公司 | High-yield Cremastra appendiculata seedling cultivation method |
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