CN104839027B - A kind of Folium Agaves variegatae bulbil abductive approach - Google Patents
A kind of Folium Agaves variegatae bulbil abductive approach Download PDFInfo
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- CN104839027B CN104839027B CN201510290075.2A CN201510290075A CN104839027B CN 104839027 B CN104839027 B CN 104839027B CN 201510290075 A CN201510290075 A CN 201510290075A CN 104839027 B CN104839027 B CN 104839027B
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Abstract
Present invention relates particularly to a kind of Folium Agaves variegatae bulbil abductive approach.The method includes drawing materials, culture medium preparation, materials disinfection process, bud inducement cultivation, enrichment culture, root induction and the plant regeneration of bulbil.It is an object of the present invention to provide a kind of Folium Agaves variegatae bulbil abductive approach, provide technical support for scale, industrialization.The present invention chooses full Folium Agaves variegatae bulbil as induced material, aseptically inducing culture 20～can sprout robust growth sprouting in 30 days；Can produce through 35～50 days enrichment cultures and breed sprout in a large number, expanding propagation coefficient is high, and generation time is short, substantially reduces growth course, seedling, transplanting survival rate can be up to more than 93% after root induction in 25 35 days；Having saved great amount of cost, after seedling is transplanted, survival rate is high, and Seedling stalwartness is tall and straight, grows fine, neat and consistent.
Description
Technical field
Present invention relates particularly to a kind of Folium Agaves variegatae bulbil abductive approach.
Background technology
Folium Agaves variegatae (Agave americana L.) has another name called the dragon tongue palm, american aloe, and for Agavaceae, Agave is perennial often
Green large-scale herbaceous plant, plant is tall and big.Leaf is the arrangement of lotus throne formula, usual 30-40 piece, 50-60 piece sometimes, large-scale, and meat drapes over one's shoulders
Needle-like is linear, long 1-2 rice, middle part width 15-20 centimetre, base portion width 10-12 centimetre, and leaf margin has dredges thorn, and there is 1 hard spine on top,
Thorn crineous is long 1.5-2.5 centimetre.Panicle is large-scale, up to 6-12 rice, multi-branched；Brightly yellowish green；Perianth tube is about 1.2
Centimetre, perianth sliver is long 2.5-3 centimetre；Stamen length is about 2 times of perianth.Capsule Long Circle, is about 5 centimetres.Because its blade is hard
Very, evergreen all the year round, brightly yellowish green, the bulbil that Post flowering inflorescence generates is few.Just bloom after typically wanting decades in original producton location, huge
Big inflorescence is high up to 7 to 8 meters, is the inflorescence grown most, white or the most hundreds of of lurid bell-shaped flower, great sight
Reward is worth, and is the common flowers of flower garden, Shelter in South China Cities and greenbelt.The Folium Agaves variegatae class plant of major part kind only opens 1 time all one's life
Flower, after spending, the ripe plant with seed is the most withered, and Folium Agaves variegatae typically can save bit by bit nutrient through a lot of years, germinates the most suddenly
Overgrowing.The most so can exhaust the energy that it is whole, the Folium Agaves variegatae after outburst soon will be withered.And some appreciable varieties are very
Difficulty is bloomed, and commonly uses plant division the most aborning, cuttage, the method for sowing are bred.
Use a large amount of high quality seedling of tissue culture technique Fast-propagation, contribute to solving scarcity of resources, for scale, industry
Change and technical support is provided.Method about Folium Agaves variegatae bulbil induced bud has no report.
Summary of the invention
It is an object of the invention to provide a kind of Folium Agaves variegatae bulbil abductive approach, provide technology to prop up for scale, industrialization
Hold.
The method of the present invention is achieved in that
A kind of Folium Agaves variegatae bulbil abductive approach, comprises the following steps:
1) method of drawing material: choosing bolting, full Folium Agaves variegatae bulbil then is material, plucks rearmounted 3-5 DEG C of refrigerator cold-storage
Save backup；
2) culture medium preparation: prepare the bud inducement culture medium of bulbil, Shoot propagation culture medium, root media respectively, cultivates
The pH value of base is 5.6-5.8, and culture medium thickness is 1.4～1.6 cm；
The bud inducement culture medium of described bulbil: MS+2.0mg L-1 6-BA+0.5mg·L-1NAA+1.0mg·L-1KT+
0.01mg·L-1 TDZ +7.0g·L-1Ag+20g·L-1Su+1.0 g·L-1Ac,
Described Shoot propagation culture medium: MS+1.0mg L-1 6-BA+0.02mg·L-1TDZ+0.3mg·L-1NAA+1.0g·
L-1Peptone+7.0g L-1Ag+20g·L-1Su +1.5 g·L-1Ac,
Described root media: 1/2MS (a great number of elements halves)+0.1mg L-1NAA+0.5-1.0mg·L-1IBA+
7.0g·L-1Ag+20g·L-1Su +1.5 g·L-1Ac；
3) materials disinfection processes: bulbil tap water is carried out surface washing, puts in saturated bleaching powder supernatant and soaks
15min, removes bulbil exoperidium after water droplet rushes 0.5-1h from the beginning, rinses 2-3 time with distilled water, use matter on superclean bench
Amount mark is 75% alcohol disinfecting 20-40s, is 0.1% mercuric chloride sterilization 12-15min with mass fraction, after sterilized water rushes 3-4 time, uses
Sterilized filter paper blots surface moisture, is placed on sterilized culture dish.
4) bud inducement of bulbil is cultivated: is seeded in the bud inducement culture medium of bulbil by the material after sterile-processed and carries out
Inducing culture；Condition of culture: culturing room's temperature is 23 ± 2 DEG C, light application time 11-12h/d, intensity of illumination is 1500-2000lx；
5) Shoot propagation is cultivated: by inducing the Multiple Buds obtained through step 4), be cut into the bulk of long 1-2cm, is seeded in bud and increases
Grow and culture medium carries out enrichment culture；Condition of culture: culturing room's temperature is 23 ± 2 DEG C, light application time 11-12h/d, intensity of illumination
For 1500-2000lx；
6) root induction is cultivated: will turn through the enrichment culture seedling individual plant with 2-4 sheet leaf, plant height 2-4cm out
Move on in root media；The condition of culture of described root culture: culturing room's temperature is 23 ± 2 DEG C, light application time 11-12h/d,
Intensity of illumination is 1500-2000lx；
7) test tube Seedling completes: when test tube Seedling length to 4-6cm is high, having the 3-5 normal root of bar form, have 2-3 sheet leaf, leaf is long
During 4-6cm, the plant regeneration completing Folium Agaves variegatae is cultivated.
The test tube Seedling described step 7) completed is transplanted: test tube Seedling is placed on nature light lower refining seedling 3-5 days, then opens
Bottle cap seedling exercising 1-2 days, gradually takes out and washes away, test tube Seedling, the culture medium being attached on root with gun-shaped forceps from culture bottle, moves
Enter in the mixed-matrix of Vermiculitum and mass ratio=1:2 humous, cover transplanting test tube Seedling, temperature control with the bell glass of band pore
System is at 20-28 DEG C, and humidity should be maintained at 65%-75%.
Above is referred to: 1, Ag is (Agar culture medium solidifying agent and the abbreviation of holder agar powder)；
2, Su is (Sugar provides plant carbon source and maintains the abbreviation of osmotic pressure sucrose)；
3, Ac is activated carbon.
Described Folium Agaves variegatae be Folium Agaves variegatae (Agave americanaL.var. marginataalba Trel)。
The present invention chooses full Folium Agaves variegatae bulbil as induced material, aseptically inducing culture 20～30 days
Can sprout robust growth sprouting；Can produce through 35～50 days enrichment cultures and breed sprout in a large number, expanding propagation coefficient is high, generation time
Short, substantially reduce growth course, seedling, transplanting survival rate can be up to more than 93% after root induction in 25-35 days；Save
Great amount of cost, after seedling is transplanted, survival rate is high, and Seedling stalwartness is tall and straight, grows fine, neat and consistent.
The remarkable advantage of the present invention:
One: existing resource can be applied to greatest extent, by animal nutrition in the short time by the inventive method
The application demand of interior solution scarce resource；
Two: sum up a set of Folium Agaves variegatae isolated culture being available for production unit application and plant regeneration technique, by this
Bright extensively application can directly improve rare kind artificial forest land's Yield and quality and afforestation level, it is achieved resource high-efficiency is trained
Educate, there is the most wide Developmental Prospect of Industrialization.Strong innovation of the present invention, with high content of technology, public welfare is strong, for promoting treasure
The research of dilute kind, utilization, have good prospect and and significance, provide theoretical foundation for developing this kind of resource
And technical support.
Accompanying drawing explanation
Fig. 1 is the bud inducement 1 of bulbil.
Fig. 2 is the bud inducement 2 of bulbil.
Fig. 3 is that Shoot propagation is cultivated.
Fig. 4 is that root induction is cultivated.
Fig. 5 is plant strain growth.
Fig. 6 is plantlet of transplant.
Detailed description of the invention
Embodiment 1
A kind of method of Folium Agaves variegatae bulbil induced bud, step is as follows:
1) method of drawing material: choose bolting then, full Folium Agaves variegatae (Agave americanaL.var.marginataalba Trel) bulbil is material；
2) culture medium preparation: prepare the bud inducement culture medium of bulbil, Shoot propagation culture medium, root media respectively, cultivates
The pH value of base is 5.6, and culture medium thickness is 1.4cm；
The bud inducement culture medium of described bulbil: MS+2.0mg L-1 6-BA+0.5mg·L-1NAA+1.0mg·L-1KT+
0.01mg·L-1 TDZ +7.0g·L-1Ag+20g·L-1Su+1.0 g·L-1Ac,
Described Shoot propagation culture medium: MS+1.0mg L-1 6-BA+0.02mg·L-1TDZ+0.3mg·L-1NAA+1.0g·
L-1Peptone+7.0g L-1Ag+20g·L-1Su +1.5 g·L-1Ac,
Described root media: 1/2MS+0.1mg L-1NAA+0.5mg·L-1IBA+ 7.0g·L-1Ag+20g·L-1Su
+1.5 g·L-1Ac；
3) materials disinfection processes: the bulbil tap water cut on plant is carried out surface washing, puts saturated bleaching powder
Supernatant soaks 15min, after water droplet rushes 0.5h from the beginning, removes bulbil exoperidium, rinse 2 times with distilled water, in ultra-clean work
It is 75% alcohol disinfecting 20s with mass fraction on platform, is 0.1% mercuric chloride sterilization 12min with mass fraction, after sterilized water rushes 3 times, uses
Sterilized filter paper blots surface moisture, is placed on sterilized culture dish.
4) bud inducement of bulbil is cultivated: is seeded in the bud inducement culture medium of bulbil by the material after sterile-processed and carries out
Inducing culture；Condition of culture: culturing room's temperature is 21 DEG C, light application time 11h/d, and intensity of illumination is 1500lx, during inducing culture
Between 40 days；
5) Shoot propagation is cultivated: by inducing the Multiple Buds obtained through step 4), be cut into the bulk of long 1-2cm, is seeded in bud and increases
Grow and culture medium carries out enrichment culture；Condition of culture: culturing room's temperature is 21 DEG C, light application time 11h/d, intensity of illumination is
1500lx, 50 days enrichment culture time；
6) root induction is cultivated: will transfer to through the enrichment culture seedling individual plant with 2 leaves, plant height 2cm out
In root media；The condition of culture of described root culture: culturing room's temperature is 21 DEG C, light application time 11h/d, intensity of illumination is
1500lx, 35 days root culture time；
7) test tube Seedling completes: when test tube Seedling length to 4cm is high, has 3 normal roots of form, has 2 leaves, during the long 4cm of leaf,
The plant regeneration completing Folium Agaves variegatae is cultivated.
8) transplant: test tube Seedling is placed on nature light lower refining seedling 3 days, then opens bottle cap seedling exercising 1 day, with gun-shaped forceps test tube Seedling
From culture bottle, gradually take out and wash away the mixed of the culture medium being attached on root, immigration Vermiculitum and mass ratio=1:2 humous
Closing in substrate, cover transplanting test tube Seedling with the bell glass of band pore, temperature controls at 20 DEG C, and humidity should be maintained at 65%, regeneration
Survival rate of plant is more than 93%.
Embodiment 2
A kind of method of Folium Agaves variegatae bulbil induced bud, step is as follows:
1) method of drawing material: choose bolting then, full Folium Agaves variegatae (Agave americanaL.var.marginataalba Trel) bulbil is material；
2) culture medium preparation: prepare the bud inducement culture medium of bulbil, Shoot propagation culture medium, root media respectively, cultivates
The pH value of base is 5.7, and culture medium thickness is 1.5 cm；
The bud inducement culture medium of described bulbil: MS+2.0mg L-1 6-BA+0.5mg·L-1NAA+1.0mg·L-1KT+
0.01mg·L-1 TDZ +7.0g·L-1Ag+20g·L-1Su+1.0 g·L-1Ac,
Described Shoot propagation culture medium: MS+1.0mg L-1 6-BA+0.02mg·L-1TDZ+0.3mg·L-1NAA+1.0g·
L-1Peptone+7.0g L-1Ag+20g·L-1Su +1.5 g·L-1Ac,
Described root media: 1/2MS+0.1mg L-1NAA+0.8mg·L-1IBA+ 7.0g·L-1Ag+20g·L-1Su
+1.5 g·L-1Ac；
3) materials disinfection processes: the bulbil tap water cut on plant is carried out surface washing, puts saturated bleaching powder
Supernatant soaks 15min, after water droplet rushes 1h from the beginning, removes bulbil exoperidium, rinse 3 times with distilled water, at superclean bench
On be 75% alcohol disinfecting 30s with mass fraction, be 0.1% mercuric chloride sterilization 13min with mass fraction, after sterilized water rushes 4 times, with disappearing
Poison filter paper blots surface moisture, is placed on sterilized culture dish.
4) bud inducement of bulbil is cultivated: is seeded in the bud inducement culture medium of bulbil by the material after sterile-processed and carries out
Inducing culture；Condition of culture: culturing room's temperature is 23 DEG C, light application time 12h/d, and intensity of illumination is 1800lx, during inducing culture
Between 30 days；
5) Shoot propagation is cultivated: by inducing the Multiple Buds obtained through step 4), be cut into the bulk of long 1-2cm, is seeded in bud and increases
Grow and culture medium carries out enrichment culture；Condition of culture: culturing room's temperature is 23 DEG C, light application time 12h/d, intensity of illumination is
1800lx, 35 days enrichment culture time；
6) root induction is cultivated: will transfer to through the enrichment culture seedling individual plant with 4 leaves, plant height 4cm out
In root media；The condition of culture of described root culture: culturing room's temperature is 23 DEG C, light application time 12h/d, intensity of illumination is
1800lx, 25 days root culture time；
7) test tube Seedling completes: when test tube Seedling length to 5cm is high, has 4 normal roots of form, has 2 leaves, during the long 5cm of leaf,
The plant regeneration completing Folium Agaves variegatae is cultivated.
The test tube Seedling described step 7) completed is transplanted: test tube Seedling is placed on nature light lower refining seedling 4 days, then opens bottle
Lid seedling exercising 2 days, gradually takes out and washes away, test tube Seedling, the culture medium being attached on root with gun-shaped forceps from culture bottle, move into trematodiasis
In the mixed-matrix of stone and mass ratio=1:2 humous, covering transplanting test tube Seedling with the bell glass of band pore, temperature controls
25 DEG C, humidity should be maintained at 70%, and regeneration plant survival rate is more than 93%.
Embodiment 3
A kind of method of Folium Agaves variegatae bulbil induced bud, step is as follows:
1) method of drawing material: choose bolting then, full Folium Agaves variegatae (Agave americanaL.var.marginataalba Trel) bulbil is material；
2) culture medium preparation: prepare the bud inducement culture medium of bulbil, Shoot propagation culture medium, root media respectively, cultivates
The pH value of base is 5.8, and culture medium thickness is 1.6 cm；
The bud inducement culture medium of described bulbil: MS+2.0mg L-1 6-BA+0.5mg·L-1NAA+1.0mg·L-1KT+
0.01mg·L-1 TDZ +7.0g·L-1Ag+20g·L-1Su+1.0 g·L-1Ac,
Described Shoot propagation culture medium: MS+1.0mg L-1 6-BA+0.02mg·L-1TDZ+0.3mg·L-1NAA+1.0g·
L-1Peptone+7.0g L-1Ag+20g·L-1Su +1.5 g·L-1Ac,
Described root media: 1/2MS+0.1mg L-1NAA+1.0mg·L-1IBA+ 7.0g·L-1Ag+20g·L-1Su
+1.5 g·L-1Ac；
3) materials disinfection processes: the bulbil tap water cut on plant is carried out surface washing, puts saturated bleaching powder
Supernatant soaks 15min, removes bulbil exoperidium after rushing 40 minutes with water droplet from the beginning, rinse 3 times with distilled water, in ultra-clean work
It is 75% alcohol disinfecting 40s with mass fraction in station, is 0.1% mercuric chloride sterilization 15min with mass fraction, after sterilized water rushes 4 times,
Blot surface moisture with sterilized filter paper, be placed on sterilized culture dish.
4) bud inducement of bulbil is cultivated: is seeded in the bud inducement culture medium of bulbil by the material after sterile-processed and carries out
Inducing culture；Condition of culture: culturing room's temperature is 25 DEG C, light application time 11h/d, and intensity of illumination is 2000lx, during inducing culture
Between 35 days；
5) Shoot propagation is cultivated: by inducing the Multiple Buds obtained through step 4), be cut into the bulk of long 1-2cm, is seeded in bud and increases
Grow and culture medium carries out enrichment culture；Condition of culture: culturing room's temperature is 25 DEG C, light application time 11h/d, intensity of illumination is
2000lx, 45 days enrichment culture time；
6) root induction is cultivated: will transfer to through the enrichment culture seedling individual plant with 3 leaves, plant height 3cm out
In root media；The condition of culture of described root culture: culturing room's temperature is 25 DEG C, light application time 11h/d, intensity of illumination is
2000lx, 30 days root culture time；
7) test tube Seedling completes: when test tube Seedling length to 6cm is high, has 5 normal roots of form, has 3 leaves, during the long 6cm of leaf,
The plant regeneration completing Folium Agaves variegatae is cultivated.
The test tube Seedling described step 7) completed is transplanted: test tube Seedling is placed on nature light lower refining seedling 5 days, then opens bottle
Lid seedling exercising 1 day, gradually takes out and washes away, test tube Seedling, the culture medium being attached on root with gun-shaped forceps from culture bottle, move into trematodiasis
In the mixed-matrix of stone and mass ratio=1:2 humous, covering transplanting test tube Seedling with the bell glass of band pore, temperature controls
28 DEG C, humidity should be maintained at 75%, and regeneration plant survival rate is more than 93%.
Claims (1)
1. a Folium Agaves variegatae bulbil abductive approach, it is characterised in that: said method comprising the steps of:
1) method of drawing material: choosing bolting, full Folium Agaves variegatae bulbil then is material, plucks rearmounted 3-5 DEG C of refrigerator cold-storage and preserves
Standby；
2) culture medium preparation: prepare the bud inducement culture medium of bulbil, Shoot propagation culture medium, root media respectively, culture medium
PH value is 5.6-5.8, and culture medium thickness is 1.4～1.6 cm；
The bud inducement culture medium of described bulbil: MS+2.0mg L-1 6-BA+0.5mg·L-1NAA+1.0mg·L-1KT+
0.01mg·L-1 TDZ +7.0g·L-1Agar+20g·L-1Sugar+1.0 g·L-1Ac,
Described Shoot propagation culture medium: MS+1.0mg L-1 6-BA+0.02mg·L-1TDZ+0.3mg·L-1NAA+1.0g·L-1Egg
White peptone+7.0g L-1Agar+20g·L-1Sugar+1.5 g·L-1Ac,
Described root media: 1/2MS+0.1mg L-1NAA+0.5-1.0mg·L-1IBA+ 7.0g·L-1Agar+20g·L- 1Sugar+1.5 g·L-1Ac；
3) materials disinfection processes: bulbil tap water is carried out surface washing, puts immersion 15min in saturated bleaching powder supernatant,
After water droplet rushes 0.5-1h from the beginning, remove bulbil exoperidium, rinse 2-3 time with distilled water, superclean bench is used mass fraction
It is 75% alcohol disinfecting 20-40s, is 0.1% mercuric chloride sterilization 12-15min with mass fraction, after sterilized water rushes 3-4 time, with sterilization filter
Paper blots surface moisture, is placed on sterilized culture dish；
4) bud inducement of bulbil is cultivated: is seeded in the bud inducement culture medium of bulbil by the material after sterile-processed and induces
Cultivate；Condition of culture: culturing room's temperature is 23 ± 2 DEG C, light application time 11-12h/d, intensity of illumination is 1500-2000lx；
5) Shoot propagation is cultivated: by inducing the Multiple Buds obtained through step 4), be cut into the bulk of long 1-2cm, is seeded in Shoot propagation training
Support in base and carry out enrichment culture；Condition of culture: culturing room's temperature is 23 ± 2 DEG C, light application time 11-12h/d, intensity of illumination is
1500-2000lx；
6) root induction is cultivated: will transfer to through the enrichment culture seedling individual plant with 2-4 sheet leaf, plant height 2-4cm out
In root media；The condition of culture of described root culture: culturing room's temperature is 23 ± 2 DEG C, light application time 11-12h/d, illumination
Intensity is 1500-2000lx；
7) test tube Seedling completes: when test tube Seedling length to 4-6cm is high, has the 3-5 normal root of bar form, has 2-3 sheet leaf, the long 4-of leaf
During 6cm, the plant regeneration completing Folium Agaves variegatae is cultivated；
The test tube Seedling described step 7) completed is transplanted: test tube Seedling is placed on nature light lower refining seedling 3-5 days, then opens bottle cap
Seedling exercising 1-2 days, gradually takes out and washes away, test tube Seedling, the culture medium being attached on root with gun-shaped forceps from culture bottle, move into trematodiasis
In the mixed-matrix of stone and mass ratio=1:2 humous, covering transplanting test tube Seedling with the bell glass of band pore, temperature controls
20-28 DEG C, humidity should be maintained at 65%-75%；
Described Folium Agaves variegatae is Folium Agaves variegatae.
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