CN104885947A - Jerusalem artichoke tissue culture propagation method - Google Patents
Jerusalem artichoke tissue culture propagation method Download PDFInfo
- Publication number
- CN104885947A CN104885947A CN201510297967.5A CN201510297967A CN104885947A CN 104885947 A CN104885947 A CN 104885947A CN 201510297967 A CN201510297967 A CN 201510297967A CN 104885947 A CN104885947 A CN 104885947A
- Authority
- CN
- China
- Prior art keywords
- jerusalem artichoke
- tissue culture
- medium
- propagation method
- explant
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention provides a jerusalem artichoke tissue culture propagation method. The jerusalem artichoke tissue culture propagation method comprises the following steps: carrying out disinfection treatment on explants; inoculating the explants to a callus induction culture medium; darkly culturing at 23-25 DEG C until calluses are formed; taking the calluses and cutting the calluses into small blocks; transferring the calluses into an adventitious bud induction culture medium; illuminating and culturing until adventitious buds grow; transferring the adventitious buds into a subculture culture medium; illuminating and culturing for 20-25 days at 23-25 DEG C; transferring subculture seedlings into a rooting culture medium and illuminating and culturing for 15-20 days at 23-25 DEG C; taking out rooted tissue culture seedlings; washing the culture medium at roots and planting into substrate; and then transplanting into a large field. According to the jerusalem artichoke tissue culture propagation method, different parts of jerusalem artichoke can be used as the explants to carry out tissue culture propagation; and the used explants are easy to obtain and the experiment operation is simple and feasible. A common MS culture medium is matched with different growth hormones to obtain a special culture medium suitable for the jerusalem artichoke tissue culture, and the tissue culture of the jerusalem artichoke can be effectively carried out, so that the jerusalem artichoke can be rapidly propagated.
Description
Technical field
The present invention relates to field of plant tissue culture, particularly relate to a kind of jerusalem artichoke tissue culture propagation method.
Background technology
Jerusalem artichoke (Helianthus tuberosus L.), also known as Jerusalem artichoke, Jerusalem artichoke, five-pointed star grass etc., is the herbaceos perennial of composite family (Compositae) Helianthus, originates in North America, import China into through Europe.Jerusalem artichoke stem tuber is of close texture, tender and crisp, edible or make pickles, and tender leaf is a kind of nutritious green fodder.The unique medicinal health effect had due to it and many processing purposes, food and agricultural organization of united state is called " 21 century, people and animals shared crop ", and DEVELOPMENT PROSPECT is very wide.Jerusalem artichoke property likes the weather of refrigerant drying, and happiness illumination avoids sweltering heat, well developed root system, has very strong drought-resistance ability, and embody the good ecological value in control of desert and water and soil conservation.
The value of jerusalem artichoke is mainly reflected on the high stem tuber of output.Jerusalem artichoke stem tuber is loose, khaki, meat white.Single fresh tuber weighs 50 ~ 70g, water content 79.8%, carbohydrate 16.6%, protein 1%, fat 1%, raw fiber 0.6%, ash content 2.8%, also containing a certain amount of thiamine, vitamin b3, niacin and ascorbic acid, contained microelements of calcium, phosphorus, iron etc. be rhizome vegetable.In addition, jerusalem artichoke is also rich in synanthrin, glucose and poly pentose and vitamin C, vitamin B1, vitamin E, potassium, magnesium and alanine etc.Jerusalem artichoke stem tuber contains and enriches synanthrin (inulin, also known as inulin), is polyfructosan, is different from starch---the polyglucose of storage in other plant corpus.
In recent years, jerusalem artichoke is with its extremely strong resistance, and significant health-care effect, purposes obtains the concern of more and more people widely.The research that jerusalem artichoke is relevant is also enriched gradually, and correlative study both at home and abroad has at present related to the many aspects such as breeding, cultivation, Physiology and biochemistry, molecular biology and comprehensive utilization.Generally jerusalem artichoke is vegetative propagation in the modes of reproduction of large Tanaka, and stem tuber long term growth, in underground, is easily subject to the infringement of multiple virus, can cause the accumulation of a large amount of virus disease, finally causes the deterioration of kind and the reduction year by year of output.Therefore setting up and improve jerusalem artichoke group culturation rapid propagating technology is the important channel addressed this problem.
Summary of the invention
Given this, be necessary to provide a kind of jerusalem artichoke tissue culture propagation method can setting up jerusalem artichoke tissue-culturing rapid propagation.
A kind of jerusalem artichoke tissue culture propagation method, comprises the steps:
To explant disinfection;
Described explant is seeded to callus inducing medium, at 23 DEG C-25 DEG C, to there is callus in light culture, wherein, described callus inducing medium is the MS medium being added with 0.8mg/L-1.6mg/L 6-benzyl aminoadenine and 0.2mg/L-0.5mg/L methyl α-naphthyl acetate;
Get described callus, be cut into fritter, be forwarded in adventitious bud induction culture base, at 23 DEG C-25 DEG C, illumination cultivation is to growing indefinite bud, wherein, described adventitious bud induction culture base is the MS medium being added with 1.0mg/L-2.0mg/L 6-benzyl aminoadenine and 0.1mg/L-0.3mg/L methyl α-naphthyl acetate;
Described indefinite bud is transferred in subculture medium, at 23 DEG C-25 DEG C, illumination cultivation 20-25 days, form Regenerated plant, wherein, described subculture medium is the MS medium being added with 1.5mg/L-2.0mg/L 6-benzyl aminoadenine;
Described Regenerated plant is proceeded in root media, at 23 DEG C-25 DEG C, illumination cultivation 15-20 days, form the plantlet in vitro of taking root, wherein, described root media is the 1/2MS medium being added with 0.2mg/L-0.6mg/L indolebutyric acid;
The plantlet in vitro of taking root described in taking-up, washes away the root media of the root of described plantlet in vitro, plants in matrix, then transplant to land for growing field crops.
Wherein in an embodiment, described explant is the stem tuber of jerusalem artichoke, stem section or blade.
Wherein in an embodiment, when described explant is stem tuber, as follows to the method for described explant disinfection:
Described stem tuber is adopted tap water, then adopts cleaning fluid to soak, cleaner with tap water, peeling, cuts bud point part, and with aseptic water washing, adopts 0.1% mercuric chloride sterilizing 7min-8min, then adopt aseptic water washing clean after stripping and slicing.
Wherein in an embodiment, when described explant is stem section, as follows to the method for described explant disinfection:
Adopt running water to clean described stem section, then adopt aseptic water washing, after segment, adopt 0.1% mercuric chloride sterilizing 5min-6min, then adopt aseptic water washing clean.
Wherein in an embodiment, when described explant is blade, as follows to the method for described explant disinfection:
Adopt running water to clean in described blade, then adopt aseptic water washing, then described blade is cut into strip, then adopt 0.1% mercuric chloride sterilizing 4min-5min, then adopt aseptic water washing clean.
Wherein in an embodiment, in described callus inducing medium, also can add 2, the 4-dichlorphenoxyacetic acids of 0.1mg/L-0.5mg/L.
The methyl α-naphthyl acetate of 0.1mg/L-0.3mg/L also can be added in described subculture medium.
Wherein in an embodiment, described in get described callus, be cut into fritter, be forwarded in adventitious bud induction culture base, at 23 DEG C-25 DEG C, illumination cultivation is in the step growing indefinite bud, light application time is 14h/d-16h/d, and intensity of illumination is 600lux-1500lux.
Wherein in an embodiment, be describedly transferred in subculture medium by described indefinite bud, at 23 DEG C-25 DEG C, illumination cultivation 20-25 days, formed in the step of Regenerated plant, light application time is 14h/d-16h/d, and intensity of illumination is 600lux-1500lux.
Wherein in an embodiment, proceed in root media by described Regenerated plant, at 23 DEG C-25 DEG C, illumination cultivation 15-20 days, formed in the step of the plantlet in vitro of taking root, light application time is 14h/d-16h/d, and intensity of illumination is 600lux-1500lux.
Wherein in an embodiment, described matrix is the mixture of the peat soil of mass ratio 1:3:1, soil and fertilizer or the turfy soil of mass ratio 1:1 and the mixture of vermiculite.
Above-mentioned jerusalem artichoke tissue culture propagation method, jerusalem artichoke different parts can be adopted to carry out group training as explant and breed, the explant used easily obtains, and experimental implementation is simple.And obtain the special culture media of suitable jerusalem artichoke tissue cultures by general MS medium different somatotropin of arranging in pairs or groups, effectively can carry out the tissue cultures of jerusalem artichoke, make jerusalem artichoke obtain Fast-propagation.
Accompanying drawing explanation
Fig. 1 is the flow chart of the jerusalem artichoke tissue culture propagation method of an embodiment;
The picture that Fig. 2 A-2C is jerusalem artichoke stem tuber, blade and stem section are seeded to callus inducing medium respectively;
The picture of the callus that Fig. 3 A-3C is jerusalem artichoke stem tuber, blade and stem section obtain respectively;
The picture of the indefinite bud that Fig. 4 A-4C is jerusalem artichoke stem tuber, blade and stem section obtain respectively;
The picture of the Regenerated plant that Fig. 5 A-5C is jerusalem artichoke stem tuber, blade and stem section obtain respectively;
Fig. 6 A-6C is the picture of the root system of jerusalem artichoke stem tuber, blade and stem section difference culture of rootage gained;
Fig. 7 A-7C is the picture of the whole plant of jerusalem artichoke stem tuber, blade and stem section difference gained.
Embodiment
In order to make object of the present invention, technical scheme and advantage more clear, as follows by reference to the accompanying drawings and embodiment, the present invention is further elaborated.Should be appreciated that specific embodiment described herein only in order to explain the present invention, be not intended to limit the present invention.
As shown in Figure 1, the jerusalem artichoke tissue culture propagation method of an embodiment, comprises the steps:
S10, to explant disinfection.
In the present embodiment, explant is the stem tuber of jerusalem artichoke, stem section or blade.
When explant is stem tuber, as follows to the method for explant disinfection:
Stem tuber is adopted tap water, then adopts cleaning fluid to soak, cleaner with tap water, peeling, cuts bud point part, and with aseptic water washing, adopts 0.1% mercuric chloride sterilizing 7min-8min, then adopt aseptic water washing clean after stripping and slicing.
Cleaning fluid can be liquid detergent water or detergent liquid etc.
When explant is stem section, as follows to the method for explant disinfection:
Adopt running water to clean stem section, then adopt aseptic water washing, after segment, adopt 0.1% mercuric chloride sterilizing 5min-6min, then adopt aseptic water washing clean.
When explant is blade, as follows to the method for explant disinfection:
Adopt running water to clean in blade, then adopt aseptic water washing, then blade is cut into strip, then adopt 0.1% mercuric chloride sterilizing 4min-5min, then adopt aseptic water washing clean.
In the present embodiment, blade can be cut into the wide strip for 1cm.
S20, explant is seeded to callus inducing medium, at 23 DEG C-25 DEG C, light culture is to occurring callus, wherein, callus inducing medium is for being added with 0.8mg/L-1.6mg/L 6-benzyl aminoadenine (6-Benzylaminopurine, 6-BA) and the MS medium of 0.2mg/L-0.5mg/L methyl α-naphthyl acetate (NAA, 1-Naphthaleneacetic acid).
In other embodiments, 2, the 4-dichlorphenoxyacetic acids (2,4-Dichlorophenoxyacetic Acid, 2,4-D) of 0.1mg/L-0.5mg/L are also added with in callus inducing medium.
The component of MS and 1/2MS medium is as shown in table 1:
Table 1
S30, get callus, be cut into fritter, be forwarded in adventitious bud induction culture base, at 23 DEG C-25 DEG C, illumination cultivation is to growing indefinite bud, wherein, adventitious bud induction culture base is the MS medium being added with 1.0mg/L-2.0mg/L 6-benzyl aminoadenine and 0.1mg/L-0.3mg/L methyl α-naphthyl acetate.
In S30, light application time can be 14h/d-16h/d, and intensity of illumination can be 600lux-1500lux.
S40, be transferred in subculture medium by indefinite bud, at 23 DEG C-25 DEG C, illumination cultivation 20-25 days, form Regenerated plant, wherein, subculture medium is the MS medium being added with 1.5mg/L-2.0mg/L 6-benzyl aminoadenine.
In other embodiments, the methyl α-naphthyl acetate of 0.1mg/L-0.3mg/L is also added with in subculture medium.
In S40, indefinite bud is transferred to before in subculture medium, cuts after indefinite bud can be waited to grow to 2cm, be transferred in subculture medium.
In S40, light application time can be 14h/d-16h/d, and intensity of illumination can be 600lux-1500lux.
S50, Regenerated plant is proceeded in root media, at 23 DEG C-25 DEG C, illumination cultivation 15-20 days, form the plantlet in vitro of taking root, wherein, root media is for being added with the 1/2MS medium of 0.2mg/L-0.6mg/L indolebutyric acid (IBA, 3-Indolebutyric acid).
In S50, Regenerated plant is proceeded to before in root media, can wait Regenerated plant grow to 3cm-4cm long time cut, proceed in root media.
In S50, light application time can be 14h/d-16h/d, and intensity of illumination can be 600lux-1500lux.
The plantlet in vitro that S60, taking-up are taken root, washes away the root media of the root of plantlet in vitro, plants in matrix, then transplant to land for growing field crops.
In the present embodiment, matrix is the mixture of the peat soil of mass ratio 1:3:1, soil and fertilizer.In other embodiments, matrix also can be the turfy soil of mass ratio 1:1 and the mixture of vermiculite.
In S60, the plantlet in vitro of taking root is planted before in matrix, plant again when the root of plantlet in vitro can be waited to be about 2cm-4cm into.Planted by plantlet in vitro after in matrix, water normal root water, shading 3 days, every day takes a breath, and then normally cultivates after 10-15 days, then transplants to land for growing field crops.
Above-mentioned jerusalem artichoke tissue culture propagation method, jerusalem artichoke different parts can be adopted to carry out group training as explant and breed, such as, using jerusalem artichoke stem tuber, blade, stem section etc. as sugarcane explants through callus induction, the explant used easily obtains, and experimental implementation is simple.And obtain the special culture media of suitable jerusalem artichoke tissue cultures by general MS medium different somatotropin of arranging in pairs or groups, effectively can carry out the tissue cultures of jerusalem artichoke, make jerusalem artichoke obtain Fast-propagation.
Be specific embodiment part below.
Embodiment 1
The explant used in the present embodiment is jerusalem artichoke stem tuber.
Explant process: jerusalem artichoke stem tuber is adopted tap water 20min, detergent liquid soaks 30min, and running water washes down, and removes the peel, be cut into about 3cm with aseptic water washing
3long block, be placed in beaker.In aseptic super-clean bench, adopt 0.1% mercuric chloride, tweezers not to stop to stir, sterilizing 7min, sterile water rinses 5-6 time repeatedly.Adopt sterilizing scalpel that jerusalem artichoke stem block is cut into 1cm again
3fritter, be seeded to callus inducing medium.It should be noted that explant chooses the position not with bud point on stem tuber.
Callus and adventitious bud inducing: in callus inducing medium, 25 DEG C, light culture 25 days.Wherein, callus inducing medium is for being added with the MS medium of 1.0mg/L 6-BA, 0.2mg/L NAA and 0.5mg/L2,4-D.In MS medium, sucrose concentration is 30g/L, agar concentration is 5g/L.。Then, be cut into small pieces by the callus after growing up and be forwarded in adventitious bud induction culture base, cultivate 25 days under intensity of illumination is 600-1500lux, light application time is 14h/d.Wherein, adventitious bud induction culture base is MS+1.0mg/L 6-BA+0.1mg/L NAA.
Squamous subculture: indefinite bud is cut after growing to 2cm, is transferred in subculture medium, and at 25 DEG C, cultivate 25 days under intensity of illumination is 600-1500lux, light application time is 14h/d.Wherein, subculture medium is MS+1.5mg/L 6-BA.
Culture of rootage: Regenerated plant grow to 4cm long time cut, proceed in root media, at 25 DEG C, under intensity of illumination is 600-1500lux cultivate 15 days, light application time is 14h/d.Wherein, root media is 1/2MS+0.4mg/L IBA.
Acclimatization and transplants: when the root of plantlet in vitro is about 3cm, carefully take out, clear water washes away root medium, plants in the matrix of culturing pot, waters normal root water, shading 3 days, and every day takes a breath, and then normally cultivates 10 days, transplants to land for growing field crops.Wherein, matrix is the mixture of the peat soil of mass ratio 1:3:1, soil and fertilizer.
Embodiment 2
The explant used in the present embodiment is jerusalem artichoke stem tuber.
Explant process: jerusalem artichoke stem tuber is adopted tap water 30min, detergent liquid soaks 30min, and running water washes down, and removes the peel, be cut into about 3cm with aseptic water washing
3long block, be placed in beaker.In aseptic super-clean bench, adopt 0.1% mercuric chloride, tweezers not to stop to stir, sterilizing 8min, sterile water rinses 5-6 time repeatedly.Adopt sterilizing scalpel that jerusalem artichoke stem block is cut into 1cm again
3fritter, be seeded to callus inducing medium.It should be noted that explant chooses the position not with bud point on stem tuber.Fig. 2 A is the picture that jerusalem artichoke stem tuber is seeded to callus inducing medium.
Callus and adventitious bud inducing: in callus inducing medium, 23 DEG C, light culture 21 days.Wherein, callus inducing medium is the MS medium being added with 1.6mg/L 6-BA and 0.4mg/L NAA.In MS medium, sucrose concentration is 30g/L, agar concentration is 5g/L.Fig. 3 A is the picture of the callus that jerusalem artichoke stem tuber obtains.Then, be cut into small pieces by the callus after growing up and be forwarded in adventitious bud induction culture base, cultivate 27 days under intensity of illumination is 600-1500lux, light application time is 15h/d.Wherein, adventitious bud induction culture base is MS+1.3mg/L 6-BA+0.3mg/L NAA.Fig. 4 A is the picture of the indefinite bud that jerusalem artichoke stem tuber obtains.
Squamous subculture: indefinite bud is cut after growing to 2cm, is transferred in subculture medium, and at 23 DEG C, cultivate 23 days under intensity of illumination is 600-1500lux, light application time is 15h/d.Wherein, subculture medium is MS+2.0mg/L 6-BA+0.3mg/L NAA.Fig. 5 A is the picture of the Regenerated plant that jerusalem artichoke stem tuber obtains.
Culture of rootage: Regenerated plant grow to 3cm long time cut, proceed in root media, at 23 DEG C, under intensity of illumination is 600-1500lux cultivate 16 days, light application time is 15h/d.Wherein, root media is 1/2MS+0.6mg/L IBA.Fig. 6 A is the picture of the root system of jerusalem artichoke stem tuber culture of rootage gained.
Acclimatization and transplants: treat that the root of plantlet in vitro is about 2cm, carefully take out, clear water washes away root medium, plants in the matrix of culturing pot, waters normal root water, shading 3 days, every day takes a breath, and then normally cultivates 15 days, transplants to land for growing field crops.Wherein, matrix is the turfy soil of mass ratio 1:1 and the mixture of vermiculite.Fig. 7 A is the picture of the whole plant of jerusalem artichoke stem tuber gained.
Embodiment 3
The explant used in the present embodiment is leaf of canada potato.
Explant process: leaf of canada potato adopted tap water and clean, with aseptic water washing, blade is cut into wide about 1cm, is about the long section of 5cm, is placed in beaker.In aseptic super-clean bench, adopt 0.1% mercuric chloride, tweezers not to stop to stir, sterilizing 5min, sterile water rinses 5-6 time repeatedly.Adopt sterilizing scalpel that blade is cut into about 1cm again
2small pieces, are seeded to callus inducing medium.
Callus and adventitious bud inducing: in callus inducing medium, 25 DEG C, light culture 25 days.Wherein, callus inducing medium is for being added with the MS medium of 1.0mg/L 6-BA, 0.2mg/L NAA and 0.5mg/L2,4-D.In MS medium, sucrose concentration is 30g/L, agar concentration is 5g/L.Then, be cut into small pieces by the callus after growing up and be forwarded in adventitious bud induction culture base, cultivate 25 days under intensity of illumination is 600-1500lux, light application time is 14h/d.Wherein, adventitious bud induction culture base is MS+1.0mg/L 6-BA+0.1mg/L NAA.
Squamous subculture: indefinite bud is cut after growing to 2cm, is transferred in subculture medium, and at 25 DEG C, cultivate 20 days under intensity of illumination is 600-1500lux, light application time is 14h/d.Wherein, subculture medium is MS+1.5mg/L 6-BA.
Culture of rootage: Regenerated plant grow to 4cm long time cut, proceed in root media, at 25 DEG C, under intensity of illumination is 600-1500lux cultivate 15 days, light application time is 14h/d.Wherein, root media is 1/2MS+0.4mg/L IBA.
Acclimatization and transplants: when the root of plantlet in vitro is about 3cm, carefully take out, clear water washes away root medium, plants in the matrix of culturing pot, waters normal root water, shading 3 days, and every day takes a breath, and then normally cultivates 15 days, transplants to land for growing field crops.Wherein, matrix is the mixture of the peat soil of mass ratio 1:3:1, soil and fertilizer.
Embodiment 4
The explant used in the present embodiment is leaf of canada potato.
Explant process: leaf of canada potato adopted tap water and clean, with aseptic water washing, blade is cut into wide about 1cm, is about the long section of 5cm, is placed in beaker.In aseptic super-clean bench, adopt 0.1% mercuric chloride, tweezers not to stop to stir, sterilizing 4min, sterile water rinses 5-6 time repeatedly.Adopt sterilizing scalpel that blade is cut into about 1cm again
2small pieces, are seeded to callus inducing medium.Fig. 2 B is the picture that leaf of canada potato is seeded to callus inducing medium.
Callus and adventitious bud inducing: in callus inducing medium, 24 DEG C, light culture 25 days.Wherein, callus inducing medium is the MS medium being added with 0.8mg/L 6-BA and 0.2mg/L NAA.In MS medium, sucrose concentration is 30g/L, agar concentration is 5g/L.Fig. 3 B is the picture of the callus that leaf of canada potato obtains.
Then, be cut into small pieces by the callus after growing up and be forwarded in adventitious bud induction culture base, cultivate 22 days under intensity of illumination is 600-1500lux, light application time is 15h/d.Wherein, adventitious bud induction culture base is MS+2.0mg/L 6-BA+0.1mg/L NAA.Fig. 4 B is the picture of the indefinite bud that leaf of canada potato obtains.
Squamous subculture: indefinite bud is cut after growing to 2cm, is transferred in subculture medium, and at 24 DEG C, cultivate 24 days under intensity of illumination is 600-1500lux, light application time is 15h/d.Wherein, subculture medium is MS+1.5mg/L 6-BA+0.1mg/L NAA.Fig. 5 B is the picture of the Regenerated plant that leaf of canada potato obtains.
Culture of rootage: Regenerated plant grow to 3cm long time cut, proceed in root media, at 24 DEG C, under intensity of illumination is 600-1500lux cultivate 17 days, light application time is 15h/d.Wherein, root media is 1/2MS+0.2mg/L IBA.Fig. 6 B is the picture of the root system of leaf of canada potato culture of rootage gained.
Acclimatization and transplants: when the root of plantlet in vitro is about 2cm, carefully take out, clear water washes away root medium, plants in the matrix of culturing pot, waters normal root water, shading 3 days, and every day takes a breath, and then normally cultivates 10 days, transplants to land for growing field crops.Wherein, matrix is the turfy soil of mass ratio 1:1 and the mixture of vermiculite.Fig. 7 B is the picture of the whole plant of leaf of canada potato gained.
Embodiment 5
The explant used in the present embodiment is jerusalem artichoke stem section.
Explant process: jerusalem artichoke stem section adopted tap water and cleans, with aseptic water washing, being cut into the long section of about 5cm, being placed in beaker.In aseptic super-clean bench, adopt 0.1% mercuric chloride, tweezers not to stop to stir, sterilizing 6min, sterile water rinses 5-6 time repeatedly, then adopts sterilizing scalpel to be cut into be about 5mm fritter, to be seeded to callus inducing medium.
Callus and adventitious bud inducing: in callus inducing medium, 24 DEG C, light culture 25 days.Wherein, callus inducing medium is the MS medium being added with 1.0mg/L 6-BA and 0.2mg/L NAA.In MS medium, sucrose concentration is 30g/L, agar concentration is 5g/L.Then, be cut into small pieces by the callus after growing up and be forwarded in adventitious bud induction culture base, cultivate 20 days under intensity of illumination is 600-1500lux, light application time is 16h/d.Wherein, adventitious bud induction culture base is MS+1.0mg/L6-BA+0.2mg/L NAA.
Squamous subculture: indefinite bud is cut after growing to 2cm, is transferred in subculture medium, and at 24 DEG C, cultivate 20 days under intensity of illumination is 600-1500lux, light application time is 16h/d.Wherein, subculture medium is MS+1.5mg/L 6-BA+0.2mg/L NAA.
Culture of rootage: Regenerated plant grow to 3cm long time cut, proceed in root media, at 24 DEG C, under intensity of illumination is 600-1500lux cultivate 20 days, light application time is 16h/d.Wherein, root media is 1/2MS+0.5mg/L IBA.
Acclimatization and transplants: treat that the root of plantlet in vitro is about 2cm, carefully take out, clear water washes away root medium, plants in the matrix of culturing pot, waters normal root water, shading 3 days, every day takes a breath, and then normally cultivates 15 days, transplants to land for growing field crops.Wherein, matrix is the turfy soil of mass ratio 1:1 and the mixture of vermiculite.
Embodiment 6
The explant used in the present embodiment is jerusalem artichoke stem section.
Explant process: jerusalem artichoke stem section adopted tap water and cleans, with aseptic water washing, being cut into the long section of about 5cm, being placed in beaker.In aseptic super-clean bench, adopt 0.1% mercuric chloride, tweezers not to stop to stir, sterilizing 5min, sterile water rinses 5-6 time repeatedly, then adopts sterilizing scalpel to be cut into be about 5mm fritter, to be seeded to callus inducing medium.Fig. 2 C is the picture that jerusalem artichoke stem section is seeded to callus inducing medium.
Callus and adventitious bud inducing: in callus inducing medium, 25 DEG C, light culture 25 days.Wherein, callus inducing medium is for being added with the MS medium of 1.0mg/L 6-BA, 0.2mg/L NAA and 0.5mg/L2,4-D.In MS medium, sucrose concentration is 30g/L, agar concentration is 5g/L.Fig. 3 C is the picture of the callus that jerusalem artichoke stem section obtains.Then, be cut into small pieces by the callus after growing up and be forwarded in adventitious bud induction culture base, cultivate 25 days under intensity of illumination is 600-1500lux, light application time is 14h/d.Wherein, adventitious bud induction culture base is MS+1.0mg/L 6-BA+0.1mg/L NAA.Fig. 4 C is the picture of the indefinite bud that jerusalem artichoke stem section obtains.
Squamous subculture: indefinite bud is cut after growing to 2cm, is transferred in subculture medium, and at 25 DEG C, cultivate 23 days under intensity of illumination is 600-1500lux, light application time is 14h/d.Wherein, subculture medium is MS+1.5mg/L 6-BA.Fig. 5 C is the picture of the Regenerated plant that jerusalem artichoke stem section obtains.
Culture of rootage: Regenerated plant grow to 4cm long time cut, proceed in root media, at 25 DEG C, under intensity of illumination is 600-1500lux cultivate 15 days, light application time is 14h/d.Wherein, root media is 1/2MS+0.4mg/L IBA.Fig. 6 C is the picture of the root system of jerusalem artichoke stem section culture of rootage gained.
Acclimatization and transplants: treat that the root of plantlet in vitro is about 3cm, carefully take out, clear water washes away root medium, plants in the matrix of culturing pot, waters normal root water, shading 3 days, every day takes a breath, and then normally cultivates 10 days, transplants to land for growing field crops.Wherein, matrix is the mixture of the peat soil of mass ratio 1:3:1, soil and fertilizer.Fig. 7 C is the picture of the whole plant of stem section gained.
The jerusalem artichoke tissue culture propagation method of embodiment 1-6 carry out the training of jerusalem artichoke group go out callus rate, bud ratio, rooting rate, survival rate please refer to table 2.As can be seen from Table 2, above-mentioned jerusalem artichoke tissue culture propagation method can carry out the tissue cultures of jerusalem artichoke effectively, makes jerusalem artichoke obtain Fast-propagation.
Table 2
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the premise without departing from the principles of the invention; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (10)
1. a jerusalem artichoke tissue culture propagation method, is characterized in that, comprises the steps:
To explant disinfection;
Described explant is seeded to callus inducing medium, at 23 DEG C-25 DEG C, to there is callus in light culture, wherein, described callus inducing medium is the MS medium being added with 0.8mg/L-1.6mg/L 6-benzyl aminoadenine and 0.2mg/L-0.5mg/L methyl α-naphthyl acetate;
Get described callus, be cut into fritter, be forwarded in adventitious bud induction culture base, at 23 DEG C-25 DEG C, illumination cultivation is to growing indefinite bud, wherein, described adventitious bud induction culture base is the MS medium being added with 1.0mg/L-2.0mg/L 6-benzyl aminoadenine and 0.1mg/L-0.3mg/L methyl α-naphthyl acetate;
Described indefinite bud is transferred in subculture medium, at 23 DEG C-25 DEG C, illumination cultivation 20-25 days, form Regenerated plant, wherein, described subculture medium is the MS medium being added with 1.5mg/L-2.0mg/L 6-benzyl aminoadenine;
Described Regenerated plant is proceeded in root media, at 23 DEG C-25 DEG C, illumination cultivation 15-20 days, form the plantlet in vitro of taking root, wherein, described root media is the 1/2MS medium being added with 0.2mg/L-0.6mg/L indolebutyric acid;
The plantlet in vitro of taking root described in taking-up, washes away the root media of the root of described plantlet in vitro, plants in matrix, then transplant to land for growing field crops.
2. jerusalem artichoke tissue culture propagation method as claimed in claim 1, is characterized in that, described explant is the stem tuber of jerusalem artichoke, stem section or blade.
3. jerusalem artichoke tissue culture propagation method as claimed in claim 2, is characterized in that, when described explant is stem tuber, as follows to the method for described explant disinfection:
Described stem tuber is adopted tap water, then adopts cleaning fluid to soak, cleaner with tap water, peeling, cuts bud point part, and with aseptic water washing, adopts 0.1% mercuric chloride sterilizing 7min-8min, then adopt aseptic water washing clean after stripping and slicing.
4. jerusalem artichoke tissue culture propagation method as claimed in claim 2, is characterized in that, when described explant is stem section, as follows to the method for described explant disinfection:
Adopt running water to clean described stem section, then adopt aseptic water washing, after segment, adopt 0.1% mercuric chloride sterilizing 5min-6min, then adopt aseptic water washing clean.
5. jerusalem artichoke tissue culture propagation method as claimed in claim 2, is characterized in that, when described explant is blade, as follows to the method for described explant disinfection:
Adopt running water to clean in described blade, then adopt aseptic water washing, then described blade is cut into strip, then adopt 0.1% mercuric chloride sterilizing 4min-5min, then adopt aseptic water washing clean.
6. jerusalem artichoke tissue culture propagation method as claimed in claim 1, is characterized in that, also can add 2, the 4-dichlorphenoxyacetic acids of 0.1mg/L-0.5mg/L in described callus inducing medium.
The methyl α-naphthyl acetate of 0.1mg/L-0.3mg/L also can be added in described subculture medium.
7. jerusalem artichoke tissue culture propagation method as claimed in claim 1, it is characterized in that, describedly get described callus, be cut into fritter, be forwarded in adventitious bud induction culture base, at 23 DEG C-25 DEG C, illumination cultivation is in the step growing indefinite bud, light application time is 14h/d-16h/d, and intensity of illumination is 600lux-1500lux.
8. jerusalem artichoke tissue culture propagation method as claimed in claim 1, is characterized in that, is describedly transferred in subculture medium by described indefinite bud, at 23 DEG C-25 DEG C, illumination cultivation 20-25 days, is formed in the step of Regenerated plant, light application time is 14h/d-16h/d, and intensity of illumination is 600lux-1500lux.
9. jerusalem artichoke tissue culture propagation method as claimed in claim 1, is characterized in that, proceed in root media by described Regenerated plant, at 23 DEG C-25 DEG C, illumination cultivation 15-20 days, is formed in the step of the plantlet in vitro of taking root, light application time is 14h/d-16h/d, and intensity of illumination is 600lux-1500lux.
10. jerusalem artichoke tissue culture propagation method as claimed in claim 1, is characterized in that, described matrix is the mixture of the peat soil of mass ratio 1:3:1, soil and fertilizer or the turfy soil of mass ratio 1:1 and the mixture of vermiculite.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510297967.5A CN104885947A (en) | 2015-06-03 | 2015-06-03 | Jerusalem artichoke tissue culture propagation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510297967.5A CN104885947A (en) | 2015-06-03 | 2015-06-03 | Jerusalem artichoke tissue culture propagation method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104885947A true CN104885947A (en) | 2015-09-09 |
Family
ID=54019271
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510297967.5A Pending CN104885947A (en) | 2015-06-03 | 2015-06-03 | Jerusalem artichoke tissue culture propagation method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104885947A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107278889A (en) * | 2017-06-21 | 2017-10-24 | 河池市智汇科技咨询有限公司 | Jerusalem artichoke adventitious bud inducing method |
| CN108770575A (en) * | 2018-05-18 | 2018-11-09 | 象州县科学技术情报研究所 | The artificial method for planting of kapok |
| CN109496571A (en) * | 2018-11-15 | 2019-03-22 | 广州市名卉景观科技发展有限公司 | A kind of cuttage breeding method of snowflake PoiyscJasbaJfouzianz |
| CN110771302A (en) * | 2019-10-25 | 2020-02-11 | 青海省农林科学院 | Method for rapidly promoting germination of jerusalem artichoke tubers after dormancy breaking |
-
2015
- 2015-06-03 CN CN201510297967.5A patent/CN104885947A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107278889A (en) * | 2017-06-21 | 2017-10-24 | 河池市智汇科技咨询有限公司 | Jerusalem artichoke adventitious bud inducing method |
| CN108770575A (en) * | 2018-05-18 | 2018-11-09 | 象州县科学技术情报研究所 | The artificial method for planting of kapok |
| CN109496571A (en) * | 2018-11-15 | 2019-03-22 | 广州市名卉景观科技发展有限公司 | A kind of cuttage breeding method of snowflake PoiyscJasbaJfouzianz |
| CN110771302A (en) * | 2019-10-25 | 2020-02-11 | 青海省农林科学院 | Method for rapidly promoting germination of jerusalem artichoke tubers after dormancy breaking |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104335903A (en) | Method for accelerating quick propagation of rhizoma bletillae | |
| CN105191805B (en) | A kind of micro-propagation method of tilia miqueliana | |
| CN105638477A (en) | Rapid propagation method for dendrobium hancockii seeds | |
| CN104106468B (en) | The quick breeding method for tissue culture of a kind of radix fici simplicissimae | |
| CN103931492A (en) | Tissue-culture rapid seedling growing method for apple rootstock M9 | |
| CN102845313A (en) | Method for quickly in-vitro actinidia kolomikta propagating | |
| CN106489740B (en) | A kind of seedling rapid propagation method using polygonatum sibiricum Redoute bulb as explant | |
| CN104067939A (en) | Tissue culture rapid propagation method of radix gentianae | |
| CN104041412A (en) | Rapid propagation method for tissue culture of Guizhou hemiboea cavaleriei | |
| CN104885947A (en) | Jerusalem artichoke tissue culture propagation method | |
| CN102884983B (en) | Method for promoting tissue culture seedling of anisetree bark to take root rapidly | |
| CN1930952A (en) | Crastinus leaf tissue culturing and fast propagation process | |
| CN103907497A (en) | Rapid cutting propagation method of test-tube plum plantlets | |
| CN103548695B (en) | A kind of meadowrueleaf corydalis root quick breeding method for tissue culture | |
| CN106165648B (en) | A kind of cercis tissue culture culture medium and cultural method | |
| CN103039362B (en) | Subculture medium for tissue culture seedling propagation of camellia oleifera abel and propagation method thereof | |
| CN104145813A (en) | Method for propagating stems of aristolochia fordiana | |
| CN109496858B (en) | Tissue culture and rapid propagation method of amomum tsao-ko | |
| CN103704135B (en) | In-vitro rapid propagation method for plantains | |
| CN103229721A (en) | Tissue culture propagation method of Gynura formosana | |
| CN105532452A (en) | Method for inducing and quickly breeding daphnemezereum seeds | |
| CN105613292A (en) | Special dendrobium officinale leaf sheath tissue culture seedling forming method | |
| CN108450328A (en) | A kind of crocodile mouth flower quick breeding method for tissue culture | |
| CN104521754A (en) | Rapid propagation method of Allium ovalifolium on high mountain | |
| CN101012448A (en) | Method of screening leymus chinensis salt-tolerant somatic mutation |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20150909 |
|
| RJ01 | Rejection of invention patent application after publication |