CN105340736B - A kind of method of swordleaf dragon tree Lax callus induction - Google Patents
A kind of method of swordleaf dragon tree Lax callus induction Download PDFInfo
- Publication number
- CN105340736B CN105340736B CN201510755473.7A CN201510755473A CN105340736B CN 105340736 B CN105340736 B CN 105340736B CN 201510755473 A CN201510755473 A CN 201510755473A CN 105340736 B CN105340736 B CN 105340736B
- Authority
- CN
- China
- Prior art keywords
- callus
- explant
- culture medium
- bud
- inoculated
- 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.)
- Expired - Fee Related
Links
Landscapes
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
A kind of method of swordleaf dragon tree Lax callus induction, comprises the following steps：(1) swordleaf dragon tree seed or bud is taken to be carried out disinfection as explant respectively；(2) explant after sterilization is placed in induction budding or bud clump in MS minimal mediums；(3) sterile bud is respectively placed in MS, B5, N6 addition BA of 6 benzyladenine 6 and methyl α-naphthyl acetate NAA culture medium and carries out culture acquisition callus；(4) callus is placed in the inducing culture of addition hormon and is cultivated to obtain more open-textured callus.The callus peak green or milky that are obtained using the method for the invention, loose, multiplication capacity is strong, can induce raised growth good Lax callus the short time, and producing swordleaf dragon tree total alkaloid to carry out cell suspension cultures provides material.
Description
Technical field
The present invention relates to a kind of plant Lax callus abductive approach, the loose callus of particularly a kind of swordleaf dragon tree
Organize abductive approach.
Background technology
Swordleaf dragon tree (Dracaena cochinchinensis (Lour) S.C.Chen) also known as wood in thousand, dragon's blood tree,
Toe dragon tree, it is vulnerable species, belongs to Chinese Second Class Key Protected Plant.Swordleaf dragon tree has very high medical value, from its resin
The dragon's blood of extraction is referred to as Sanguis Draxonis, is China's tradition rare traditional Chinese medicine, in existing more than 1500 usage history in China.According to
《Compendium of Materia Medica》Record, Resina Draconis is warm-natured, flat, sweet, salty, nontoxic, enters blood system, and return lung, spleen, kidney three pass through, have it is promoting blood circulation and removing blood stasis,
The remarkable efficacies such as swelling and pain relieving, astringing to arrest bleeding, softening and resolving hard mass, myogenic sore, there are the good reputation of " promoting blood circulation panacea ", main distribution clouds
The ground such as south, Guangxi, Hainan.
In recent years, because its demand is greatly along with fancy price causes people to implement swordleaf dragon tree wild resource
It is irrational it is predatory fell, swordleaf dragon tree wild resource is increasingly exhausted, endangered, by many countries in the world
It is classified as rare endangered plants.Therefore it is particularly important tissue-culturing rapid propagation to be carried out to it using biotechnology.To swordleaf dragon tree
Research is mainly in the research of pharmaceutical chemistry composition, tissue cultures etc., and there has been no research in terms of cell suspension cultures at present
Report.Lax callus is easily fragmentary, and multiplication capacity is vigorous, is preserved by long-term subculture without losing embryo, is advantageous to be cured
Injured tissue establishes excellent suspension system, and can therefrom isolate the protoplast of totipotency when needed.By probing into difference
Influence of the hormone to swordleaf dragon tree callus, fast-growth, the maximum hormone ratio of loose type callus are filtered out, is
Carry out cell suspension cultures production swordleaf dragon tree total alkaloid and material is provided, provided for protection and reasonable development swordleaf dragon tree
Technical method.
The content of the invention
It is an object of the invention to provide the method that a kind of Lax callus of swordleaf dragon tree induces, it being capable of the short time
Produce the vigorous Lax callus of multiplication capacity, provide material to carry out cell suspension cultures production swordleaf dragon tree total alkaloid
Material.
The present invention reaches above-mentioned purpose by the following technical programs：A kind of Lax callus induction of swordleaf dragon tree
Method, comprise the following steps：
(1) sterilization of explant：Liquid detergent aqueous cleaning explant surface smut is first used, is placed in beaker and carries out flowing water
5~8min is rinsed, is then moved on superclean bench, seed and bud are soaked into 30s, aseptic water washing 1 with 75v/v% alcohol
Time, then use 0.1v/v%HgC12Immersion, the time is respectively 8,12,15min, sterilized water soaks 3 times, blotted with aseptic filter paper outer
It is inoculated with after implant surface moisture, wherein sterilized water is the distilled water through autoclave sterilization；The explant is full kind
Son or seed are seeded in the bud sprouted in river sand；
(2) acquisition of aseptic seedling：The explant that step (1) obtains is inoculated in MS inducing cultures, in cultivation temperature
For 25 ± 2 DEG C, 20~30 μm of ol/m of intensity of illumination2S, 8~15d of culture just starts under conditions of light application time is 12h/d
Bud, aseptic seedling is then grown, 0.5mg/L 6-benzyladenine 6-BA, 0.1mg/L naphthalene are added in the MS inducing cultures
Acetic acid NAA, 30g/L sucrose, 5g/L agar, the pH value of culture medium is 5.8；
(3) inducing effect of the culture medium to callus：The aseptic seedling obtained in step (2) is inoculated in MS, B5, N6 tri-
It is 25 ± 2 DEG C in cultivation temperature in kind minimal medium, 20~30 μm of ol/m of intensity of illumination2S, light application time are 12h/d's
Under the conditions of cultivate 25~30d, gradually produce Lax callus, added respectively in described tri- kinds of minimal mediums of MS, B5, N6
0.5mg/L 6-benzyladenine 6-BA, 0.2mg/L methyl α-naphthyl acetate NAA, 30g/L sucrose, 5g/L agar, the pH of culture medium
It is worth for 5.8；
(4) subculture effect of the culture medium to callus：The callus obtained in step (3) is placed in MS squamous subcultures
In base, in 25 ± 2 DEG C of cultivation temperature, 20~30 μm of ol/m of intensity of illumination2S, light application time are cultivated under conditions of being 10h/d
10d, cut ends are gradually expanded, and form more loose callus, 0.1~2.0mg/L is added in the MS subculture mediums
6-benzyladenine 6-BA, 0.5~3.0mg/L 2,4- dichlorphenoxyacetic acids 2,4-D, 30g/L sucrose, 5g/L agar,
The pH value of culture medium is 5.8.
The present invention's has the prominent advantages that：
(1) by taking swordleaf dragon tree seed or bud to carry out callus induction, can cultivate in a short time a large amount of
The swordleaf dragon tree Lax callus to grow fine, establish suspension system for swordleaf dragon tree and excellent material is provided.
(2) the present invention relates to mercuric chloride, alcohol, MS, B5, N6 minimal medium and 6-benzyladenine 6-BA, methyl α-naphthyl acetate NAA
With 2,4 dichlorophenoxyacetic acid 2,4-D.MS has higher inorganic salt concentration, using the teaching of the invention it is possible to provide the mineral battalion needed for tissue growth
Support, so as to accelerate the growth of callus.B5 medium is mainly characterized by containing relatively low ammonium, and this nutritional ingredient of ammonium is to training
Foster base plays the role of to suppress growth.N6 culture medium features are KNO3(NH4)2SO4Content is high.
(3) chemicals such as disinfectant, hormone is cheap used in, and experimentation cost is low.
(4) the swordleaf dragon tree callus tissue culture 40d inductivities obtained using cultural method of the present invention are reached
More than 85%, caused callus growth is fast, most of for peak green, the loose type callus of milky white granules shape.
Embodiment
Technical scheme is further illustrated with reference to embodiment.
Embodiment 1
One example of the method for the Lax callus induction of swordleaf dragon tree of the present invention, including following step
Suddenly：
(1) sterilization of explant：Liquid detergent aqueous cleaning explant surface smut is first used, is placed in beaker and carries out flowing water
5~8min is rinsed, is then moved on superclean bench, seed and bud are soaked into 30s, aseptic water washing 1 with 75v/v% alcohol
Time, then use 0.1v/v%HgC12Immersion, the time is respectively 8,12,15min, sterilized water soaks 3 times.Blotted with aseptic filter paper outer
It is inoculated with after implant surface moisture, wherein sterilized water is the distilled water through autoclave sterilization；The explant is full kind
Son or seed are seeded in the bud sprouted in river sand；
(2) acquisition of aseptic seedling：The explant that step (1) obtains is inoculated in MS inducing cultures respectively, cultivated
Temperature is 25 ± 2 DEG C, 20~30 μm of ol/m of intensity of illumination2S, light application time are cultivated 8~15d under conditions of being 12h/d, just opened
Begin to sprout, then grow aseptic seedling.Be additionally added in the MS inducing cultures 0.5mg/L 6-benzyladenine 6-BA,
0.1mg/L methyl α-naphthyl acetate NAA, 30g/L sucrose, 5g/L agar, the pH value of culture medium is 5.8；
(3) inducing effect of the culture medium to callus：By the aseptic seedling obtained in step (2) be inoculated in respectively MS, B5,
It is 25 ± 2 DEG C in cultivation temperature in tri- kinds of minimal mediums of N6,20~30 μm of ol/m of intensity of illumination2S, light application time are
25~30d is cultivated under conditions of 12h/d, gradually produces Lax callus.It is additionally added in MS, B5, N6 minimal medium
0.5mg/L 6-benzyladenine 6-BA, 0.2mg/L methyl α-naphthyl acetate NAA, 30g/L sucrose, 5g/L agar, the pH of culture medium
It is worth for 5.8；Tri- kinds of minimal medium callus induction rates of MS, B5, N6 be respectively for 62.9%, 57.1%, 51.4%, MS bases
Callus growth caused by being cultivated in basal culture medium is vigorous, short texture, is secondly B5 medium, using N6 as culture medium callus
Tissue inductivity is minimum, and color is pale yellow, callus quality is close, melting brown rate highest；
(4) subculture effect of the culture medium to callus：The callus obtained in step (3) is placed in MS squamous subcultures
It is 25 ± 2 DEG C in cultivation temperature in base, 20~30 μm of ol/m of intensity of illumination2S, light application time are cultivated under conditions of being 12h/d
After 10d, cut ends are gradually expanded, and form more loose callus, it is additionally added 0.1 in the MS subculture mediums~
2.0mg/L 6-benzyladenine 6-BA, 0.5mg/L 2,4- dichlorphenoxyacetic acids 2,4-D, 30g/L sucrose, 5g/L fine jade
Fat, the pH value of culture medium are 5.8, and callus induction rate is up to 77.1%, and color is yellowish or light green color, short texture.
Embodiment 2
(1) sterilization of explant：Liquid detergent aqueous cleaning explant surface smut is first used, is placed in beaker and carries out flowing water
5~8min is rinsed, is then moved on superclean bench, seed and bud are soaked into 30s, aseptic water washing 1 with 75v/v% alcohol
Time, then use 0.1v/v%HgC12Immersion, the time is respectively 8,12,15min, sterilized water soaks 3 times.Blotted with aseptic filter paper outer
It is inoculated with after implant surface moisture, wherein sterilized water is the distilled water through autoclave sterilization；The explant is full kind
Son or seed are seeded in the bud sprouted in river sand；
(2) acquisition of aseptic seedling：The explant that step (1) obtains is inoculated in MS inducing cultures respectively, cultivated
Temperature is 25 ± 2 DEG C, 20~30 μm of ol/m of intensity of illumination2S, light application time are cultivated under conditions of being 12h/d, cultivate 8~15d
Just start to sprout, then grow aseptic seedling.The explant inducing culture is using MS culture mediums as minimal medium, is additionally added
0.5mg/L 6-benzyladenine 6-BA, 0.1mg/L methyl α-naphthyl acetate NAA, 30g/L sucrose, 5g/L agar, the pH of culture medium
It is worth for 5.8；
(3) inducing effect of the culture medium to callus：It is that explant is inoculated in MS by the sterile bud obtained in step (2)
It is 25 ± 2 DEG C in cultivation temperature in minimal medium, 20~30 μm of ol/m of intensity of illumination2S, light application time are 12h/d bar
Cultivated under part, 25~30d gradually produces Lax callus.The callus inducing medium is respectively that MS is cultivated substantially
Base, it is additionally added 0.5mg/L 6-benzyladenine 6-BA, 0.2mg/L methyl α-naphthyl acetate NAA, 30g/L sucrose, 5g/L agar, training
The pH value for supporting base is 5.8；
(4) subculture effect of the culture medium to callus：The callus obtained in step (3) is placed in MS squamous subcultures
In base, in 25 ± 2 DEG C of cultivation temperature, 20~30 μm of ol/m of intensity of illumination2S, light application time are cultivated under conditions of being 10h/d,
After being inoculated with 10d, cut ends are gradually expanded, and form more loose callus, the callus subculture medium is with MS
Culture medium is minimal medium, is additionally added 0.5mg/L 6-benzyladenine 6-BA, 0.1mg/L 2,4- dichlorphenoxyacetic acids
2,4-D, 30g/L sucrose, 5g/L agar, the pH value of culture medium is 5.8, and callus induction rate 65.7%, color is light green
Color, close structure.
Embodiment 3
(1) sterilization of explant：Liquid detergent aqueous cleaning explant surface smut is first used, is placed in beaker and carries out flowing water
5~8min is rinsed, is then moved on superclean bench, seed and bud are soaked into 30s, aseptic water washing 1 with 75v/v% alcohol
Time, then use 0.1v/v%HgC12Immersion, the time is respectively 8,12,15min, sterilized water soaks 3 times.Blotted with aseptic filter paper outer
It is inoculated with after implant surface moisture, wherein sterilized water is the distilled water through autoclave sterilization；The explant is full kind
Son or seed are seeded in the bud sprouted in river sand；
(2) acquisition of aseptic seedling：The explant that step (1) obtains is inoculated in MS inducing cultures respectively, cultivated
Temperature is 25 ± 2 DEG C, 20~30 μm of ol/m of intensity of illumination2S, light application time are cultivated under conditions of being 12h/d, cultivate 8~15d
Just start to sprout, then grow aseptic seedling.The explant inducing culture is using MS culture mediums as minimal medium, is additionally added
0.5mg/L 6-benzyladenine 6-BA, 0.1mg/L methyl α-naphthyl acetate NAA, 30g/L sucrose, 5g/L agar, the pH of culture medium
It is worth for 5.8；
(3) inducing effect of the culture medium to callus：It is that explant is inoculated in MS by the sterile bud obtained in step (2)
It is 25 ± 2 DEG C in cultivation temperature in minimal medium, 20~30 μm of ol/m of intensity of illumination2S, light application time are 12h/d bar
Cultivated under part, 25~30d gradually produces Lax callus.The callus inducing medium is respectively that MS is cultivated substantially
Base, it is additionally added 0.5mg/L 6-benzyladenine 6-BA, 0.2mg/L methyl α-naphthyl acetate NAA, 30g/L sucrose, 5g/L agar, training
The pH value for supporting base is 5.8；
(4) subculture effect of the culture medium to callus：The callus obtained in step (3) is placed in MS squamous subcultures
In base, in 25 ± 2 DEG C of cultivation temperature, 20~30 μm of ol/m of intensity of illumination2S, light application time are cultivated under conditions of being 10h/d,
After being inoculated with 10d, cut ends are gradually expanded, and form more loose callus, the callus subculture medium is with MS
Culture medium is minimal medium, is additionally added 0.5mg/L 6-benzyladenine 6-BA, 1.0mg/L 2,4- dichlorphenoxyacetic acids
2,4-D, 30g/L sucrose, 5g/L agar, the pH value of culture medium is 5.8, callus induction rate 80.0%, colors green,
Short texture.
Embodiment 4
(1) sterilization of explant：Liquid detergent aqueous cleaning explant surface smut is first used, is placed in beaker and carries out flowing water
5~8min is rinsed, is then moved on superclean bench, seed and bud are soaked into 30s, aseptic water washing 1 with 75v/v% alcohol
Time, then use 0.1v/v%HgC12Immersion, the time is respectively 8,12,15min, sterilized water soaks 3 times.Blotted with aseptic filter paper outer
It is inoculated with after implant surface moisture, wherein sterilized water is the distilled water through autoclave sterilization；The explant is full kind
Son or seed are seeded in the bud sprouted in river sand；
(2) acquisition of aseptic seedling：The explant that step (1) obtains is inoculated in MS inducing cultures respectively, cultivated
Temperature is 25 ± 2 DEG C, 20~30 μm of ol/m of intensity of illumination2S, light application time are cultivated under conditions of being 12h/d, cultivate 8~15d
Just start to sprout, then grow aseptic seedling.The explant inducing culture is using MS culture mediums as minimal medium, is additionally added
0.5mg/L 6-benzyladenine 6-BA, 0.1mg/L methyl α-naphthyl acetate NAA, 30g/L sucrose, 5g/L agar, the pH of culture medium
It is worth for 5.8；
(3) inducing effect of the culture medium to callus：It is that explant is inoculated in MS by the sterile bud obtained in step (2)
It is 25 ± 2 DEG C in cultivation temperature in minimal medium, 20~30 μm of ol/m of intensity of illumination2S, light application time are 12h/d bar
Cultivated under part, 25~30d gradually produces Lax callus.The callus inducing medium is respectively that MS is cultivated substantially
Base, it is additionally added 0.5mg/L 6-benzyladenine 6-BA, 0.2mg/L methyl α-naphthyl acetate NAA, 30g/L sucrose, 5g/L agar, training
The pH value for supporting base is 5.8；
(4) subculture effect of the culture medium to callus：The callus obtained in step (3) is placed in MS squamous subcultures
In base, in 25 ± 2 DEG C of cultivation temperature, 20~30 μm of ol/m of intensity of illumination2S, light application time are cultivated under conditions of being 10h/d,
After being inoculated with 10d, cut ends are gradually expanded, and form more loose callus, the callus subculture medium is with MS
Culture medium is minimal medium, is additionally added 0.5mg/L 6-benzyladenine 6-BA, 1.5mg/L 2,4- dichlorphenoxyacetic acids
2,4-D, 30g/L sucrose, 5g/L agar, the pH value of culture medium is 5.8, and callus induction rate 88.6%, color is milky white
Color or peak green, structure are more loose.
Embodiment 5
(1) sterilization of explant：Liquid detergent aqueous cleaning explant surface smut is first used, is placed in beaker and carries out flowing water
5~8min is rinsed, is then moved on superclean bench, seed and bud are soaked into 30s, aseptic water washing 1 with 75v/v% alcohol
Time, then use 0.1v/v%HgC12Immersion, the time is respectively 8,12,15min, sterilized water soaks 3 times.Blotted with aseptic filter paper outer
It is inoculated with after implant surface moisture, wherein sterilized water is the distilled water through autoclave sterilization；The explant is full kind
Son or seed are seeded in the bud sprouted in river sand；
(2) acquisition of aseptic seedling：The explant that step (1) obtains is inoculated in MS inducing cultures respectively, cultivated
Temperature is 25 ± 2 DEG C, 20~30 μm of ol/m of intensity of illumination2S, light application time are cultivated under conditions of being 12h/d, cultivate 8~15d
Just start to sprout, then grow aseptic seedling.The explant inducing culture is using MS culture mediums as minimal medium, is additionally added
0.5mg/L 6-benzyladenine 6-BA, 0.1mg/L methyl α-naphthyl acetate NAA, 30g/L sucrose, 5g/L agar, the pH of culture medium
It is worth for 5.8；
(3) inducing effect of the culture medium to callus：It is that explant is inoculated in MS by the sterile bud obtained in step (2)
It is 25 ± 2 DEG C in cultivation temperature in minimal medium, 20~30 μm of ol/m of intensity of illumination2S, light application time are 12h/d bar
Cultivated under part, 25~30d gradually produces Lax callus.The callus inducing medium is respectively that MS is cultivated substantially
Base, it is additionally added 0.5mg/L 6-benzyladenine 6-BA, 0.2mg/L methyl α-naphthyl acetate NAA, 30g/L sucrose, 5g/L agar, training
The pH value for supporting base is 5.8；
(4) subculture effect of the culture medium to callus：The callus obtained in step (3) is placed in MS squamous subcultures
In base, in 25 ± 2 DEG C of cultivation temperature, 20~30 μm of ol/m of intensity of illumination2S, light application time are cultivated under conditions of being 10h/d,
After being inoculated with 10d, cut ends are gradually expanded, and form more loose callus, the callus subculture medium is with MS
Culture medium is minimal medium, is additionally added 0.5mg/L 6-benzyladenine 6-BA, 2.0mg/L 2,4- dichlorphenoxyacetic acids
2,4-D, 30g/L sucrose, 5g/L agar, the pH value of culture medium is 5.8, and callus induction rate 85.7%, color is light green
Color or milky, structure are most loose.
Embodiment 6
(1) sterilization of explant：Liquid detergent aqueous cleaning explant surface smut is first used, is placed in beaker and carries out flowing water
5~8min is rinsed, is then moved on superclean bench, seed and bud are soaked into 30s, aseptic water washing 1 with 75v/v% alcohol
Time, then use 0.1v/v%HgC12Immersion, the time is respectively 8,12,15min, sterilized water soaks 3 times.Blotted with aseptic filter paper outer
It is inoculated with after implant surface moisture, wherein sterilized water is the distilled water through autoclave sterilization；The explant is full kind
Son or seed are seeded in the bud sprouted in river sand；
(2) acquisition of aseptic seedling：The explant that step (1) obtains is inoculated in MS inducing cultures respectively, cultivated
Temperature is 25 ± 2 DEG C, 20~30 μm of ol/m of intensity of illumination2S, light application time are cultivated under conditions of being 12h/d, cultivate 8~15d
Just start to sprout, then grow aseptic seedling.The explant inducing culture is using MS culture mediums as minimal medium, is additionally added
0.5mg/L 6-benzyladenine 6-BA, 0.1mg/L methyl α-naphthyl acetate NAA, 30g/L sucrose, 5g/L agar, the pH of culture medium
It is worth for 5.8；
(3) inducing effect of the culture medium to callus：It is that explant is inoculated in MS by the sterile bud obtained in step (2)
It is 25 ± 2 DEG C in cultivation temperature in minimal medium, 20~30 μm of ol/m of intensity of illumination2S, light application time are 12h/d bar
Cultivated under part, 25~30d gradually produces Lax callus.The callus inducing medium is respectively that MS is cultivated substantially
Base, it is additionally added 0.5mg/L 6-benzyladenine 6-BA, 0.2mg/L methyl α-naphthyl acetate NAA, 30g/L sucrose, 5g/L agar, training
The pH value for supporting base is 5.8；
(4) subculture effect of the culture medium to callus：The callus obtained in step (3) is placed in MS squamous subcultures
In base, in 25 ± 2 DEG C of cultivation temperature, 20~30 μm of ol/m of intensity of illumination2S, light application time are cultivated under conditions of being 10h/d,
After being inoculated with 10d, cut ends are gradually expanded, and form more loose callus, the callus subculture medium is with MS
Culture medium is minimal medium, is additionally added 0.5mg/L 6-benzyladenine 6-BA, 3.0mg/L 2,4- dichlorphenoxyacetic acids
2,4-D, 30g/L sucrose, 5g/L agar, the pH value of culture medium is 5.8, callus induction rate 82.9%, colors green
Or faint yellow, short texture.
Claims (1)
- A kind of 1. method of the Lax callus induction of swordleaf dragon tree, it is characterised in that：This method comprises the steps of：(1) sterilization of explant：Liquid detergent aqueous cleaning explant surface smut is first used, is placed in progress flowing water flushing in beaker 5~8min, then move on superclean bench, seed and bud are soaked into 30s with 75%v/v alcohol, aseptic water washing 1 time, then With 0.1%v/v HgC12Immersion, the time is respectively 8,12,15min, sterilized water soaked 3 times, and explant is blotted with aseptic filter paper It is inoculated with after surface moisture, wherein sterilized water is the distilled water through autoclave sterilization；The explant be full seed or Seed is seeded in the bud sprouted in river sand；(2) acquisition of aseptic seedling：The explant that step (1) obtains is inoculated in MS inducing cultures, is 25 in cultivation temperature ± 2 DEG C, 20~30 μm of ol/m of intensity of illumination2S, 8~15d of culture just starts to sprout under conditions of light application time is 12h/d, with After grow aseptic seedling, 0.5mg/L 6-benzyladenine 6-BA, 0.1mg/L methyl α-naphthyl acetate are added in the MS inducing cultures NAA, 30g/L sucrose, 5g/L agar, the pH value of culture medium is 5.8；(3) inducing effect of the culture medium to callus：The sterile bud obtained in step (2) is inoculated in MS minimal mediums In, it is 25 ± 2 DEG C in cultivation temperature, 20~30 μm of ol/m of intensity of illumination2S, light application time cultivate 25 under conditions of being 12h/d ~30d, gradually produces Lax callus, added in the MS minimal mediums 0.5mg/L 6-benzyladenine 6-BA, 0.2mg/L methyl α-naphthyl acetate NAA, 30g/L sucrose, 5g/L agar, the pH value of culture medium is 5.8；(4) subculture effect of the culture medium to callus：The callus obtained in step (3) is placed in MS subculture mediums In, in 25 ± 2 DEG C of cultivation temperature, 20~30 μm of ol/m of intensity of illumination2S, light application time cultivate 10d under conditions of being 10h/d, Cut ends are gradually expanded, and form more loose callus, and 0.5mg/L 6- benzyl glands are added in the MS subculture mediums Purine 6-BA, 1.5~2.0mg/L 2,4- dichlorphenoxyacetic acids 2,4-D, 30g/L sucrose, 5g/L agar, the pH of culture medium It is worth for 5.8.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510755473.7A CN105340736B (en) | 2015-11-09 | 2015-11-09 | A kind of method of swordleaf dragon tree Lax callus induction |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510755473.7A CN105340736B (en) | 2015-11-09 | 2015-11-09 | A kind of method of swordleaf dragon tree Lax callus induction |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105340736A CN105340736A (en) | 2016-02-24 |
| CN105340736B true CN105340736B (en) | 2017-12-19 |
Family
ID=55317026
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510755473.7A Expired - Fee Related CN105340736B (en) | 2015-11-09 | 2015-11-09 | A kind of method of swordleaf dragon tree Lax callus induction |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105340736B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108419672B (en) * | 2018-01-26 | 2021-09-28 | 长江大学 | Method for calluses of dracaena cochinchinensis to form seedlings |
| CN114208678B (en) * | 2021-12-31 | 2023-03-14 | 海南大学 | Inducing method of cotyledon callus of mother-of-law fern |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| USPP4035P (en) * | 1976-05-14 | 1977-04-05 | Cerro Gordo, Inc. | Dracaena colorama |
| CN1316876C (en) * | 2004-12-27 | 2007-05-23 | 中国热带农业科学院热带生物技术研究所 | Method for cultivating and producing dragon's blood using dragon dracaena tissue |
| CN103548694B (en) * | 2013-11-04 | 2015-04-15 | 广西壮族自治区药用植物园 | Tissue culture and rapid propagation method for dracaena cochinchinensis |
| CN104472362B (en) * | 2014-12-05 | 2016-03-30 | 广西壮族自治区药用植物园 | A kind of method of swordleaf dragon tree callus induction and anti-browning |
-
2015
- 2015-11-09 CN CN201510755473.7A patent/CN105340736B/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN105340736A (en) | 2016-02-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105325301A (en) | Tissue culture method achieving two-step seedling and rapid propagation of chiritopsis cordifolia | |
| CN106718890B (en) | A kind of method for tissue culture suitable for Chinese herbal medicine murraya paniculataJack seedling | |
| CN103548694B (en) | Tissue culture and rapid propagation method for dracaena cochinchinensis | |
| CN103348920B (en) | Rapid propagation method for high quality seedlings of Kyara | |
| CN104206281B (en) | The method for tissue culture that Radix clematidis floridae gracefulness is purple | |
| CN104813939A (en) | Method for constructing lotus regeneration system | |
| CN106665232A (en) | Cutting seedling method for Chinese rose trees | |
| CN104067939A (en) | Tissue culture rapid propagation method of radix gentianae | |
| CN104041412A (en) | Rapid propagation method for tissue culture of Guizhou hemiboea cavaleriei | |
| CN103477981A (en) | Acer palmatum Seiryu tissue culture propagation process | |
| CN106665243A (en) | Cutting seedling culturing method for plum blossoms | |
| CN104472355A (en) | Method for rapidly reproduction overwintering bud of epimedium sagittatum | |
| CN106665057A (en) | Cutting seedling raising method for Arabian jasmine flowers | |
| CN104996298A (en) | Method for cultivating polygala fallax hemsl tissue culture seedlings based on multiple internodal stem segments integration | |
| CN107182791A (en) | A kind of bletilla striata tissue culture and rapid propagation method | |
| CN105475139B (en) | A kind of tissue culture propagation method of mao of leaf venushair fern | |
| CN104686329A (en) | Tissue culture method for Eucommia ulmoides Oliv. | |
| CN105340736B (en) | A kind of method of swordleaf dragon tree Lax callus induction | |
| CN102870683A (en) | Microbody propagation expanding method of aquilaria malaccensis | |
| CN105191792A (en) | Intermediate propagation method of vernonia amygdalina | |
| CN104488722B (en) | A kind of quick breeding method for tissue culture of South America crutch flower | |
| CN106359101A (en) | Tissue culture and rapid propagation method of ficus deltoidea | |
| CN103548695B (en) | A kind of meadowrueleaf corydalis root quick breeding method for tissue culture | |
| CN104472362B (en) | A kind of method of swordleaf dragon tree callus induction and anti-browning | |
| CN105010141A (en) | Anthurium rapid propagation method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20171219 Termination date: 20211109 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |