CN107047316A - A kind of iris tissue culture method and culture medium - Google Patents
A kind of iris tissue culture method and culture medium Download PDFInfo
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- CN107047316A CN107047316A CN201710417498.5A CN201710417498A CN107047316A CN 107047316 A CN107047316 A CN 107047316A CN 201710417498 A CN201710417498 A CN 201710417498A CN 107047316 A CN107047316 A CN 107047316A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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Abstract
The invention discloses a kind of iris tissue culture method, seedling culture including Multiplying culture liquid, compound a great number of elements, compound micro substance, inositol, 6 benzyl purines, 2,4 fenacs, indolebutyric acid, mashed potatoes, sucrose are included in Multiplying culture liquid；The strong seedling culture of strong seedling culture liquid, strong seedling culture liquid includes compound a great number of elements, compound micro substance, inositol, peptone, activated carbon, citric acid, ammonium nitrate, potassium nitrate, mashed potatoes, banana puree, sucrose；The adaptability culture of grown cultures liquid, grown cultures liquid includes compound a great number of elements, compound micro substance, inositol, indolebutyric acid, peptone, activated carbon, citric acid, ammonium nitrate, potassium nitrate, a great number of elements auxiliary agent, mashed potatoes, banana puree, sucrose.The invention also discloses the culture medium of culture iris, germination percentage improves 6%, and seedling, which grows up to, to be shortened 10 days, and seedling percent is 100%, and strong sprout grows up to the time and shortened 14 days.
Description
Technical field
The invention belongs to field of plant tissue culture technique, and in particular to a kind of iris tissue culture method and culture
Base.
Background technology
Iris (Phalaenopsis aphrodite Rchb.F.) is orchid family Phalaenopsis, originates in hylaeion hypotropicum
Area, for growing nonparasitically upon another plant property orchid.The thick aerial root of iris white is exposed at around blade, except the work with nutrient in absorption air
With outer, also grow and photosynthesis.Time in the new year, Phalaenopsis plants extract long bennet out from axil, and output shape
Flower as butterfly dances in the air, the deep favor by flower fans have the title of " cattleya queen consort ".
In order to meet the demand of flowers city industry, traditional iris planting patterns has been difficult to meet the market demand, because
The most frequently used implantation methods of this current iris be using its tissue carry out vegetative propagation, so can either amount reproduction plant,
It can be that filial generation iris meets maternal phenotypic character to have, and application prospect is good.
The orchid class loading culture such as existing iris is usually commercially available " spending precious No. one ", wherein main component with nutrient solution
For nitrogen, phosphorus, potassium element, still " spend precious No. one " and be primarily directed to the work of promotion root growth when orchid cuttage, grafting, transplanting
With, it is difficult to meet the fast-growth of whole iris growth cycle.
The content of the invention
A kind of iris tissue culture method and culture medium that the present invention is provided, for the need of iris different growth phases
Ask and design different culture mediums, can shorten whole iris growth cycle by this method.
A kind of iris tissue culture method that the present invention is provided, comprises the following steps：
Step 1, after the pedicel axillary buds of iris are carried out disinfection, otch is carried out to pedicel axillary buds, propagation is then seeded to
In nutrient solution, cultivate 12-18 days, temperature is 20-25 DEG C, intensity of illumination 1500-2000Lux obtains iris seedling；
The formula of the Multiplying culture liquid is as follows, and every liter of Multiplying culture liquid is made up of following components：Compound a great number of elements 2g,
Compound micro substance 1.3-1.5g, inositol 120-130mg, 6- benzyl purine 2-3mg, 2,4 dichloro benzene acetic acid 2-3mg, indoles fourth
Sour 0.4-0.6mg, mashed potatoes 100g, sucrose 20-25g, agar powder 5g, surplus are water, and the pH of Multiplying culture liquid is 5.8；
Step 2, iris seedling is transferred in strong seedling culture liquid, cultivates 22-24 days, obtain iris strong sprout；
The formula of the strong seedling culture liquid is as follows, and every liter of strong seedling culture liquid is made up of following components：Compound a great number of elements 1g,
Compound micro substance 1.3-1.5g, inositol 100mg, peptone 2-3g, activated carbon 1-2g, citric acid 35-45mg, ammonium nitrate 620-
660mg, potassium nitrate 170-180mg, mashed potatoes 40g, banana puree 35-45g, sucrose 20-25g, agar powder 5g, surplus are water, and
The pH of strong seedling culture liquid is 6.0；
Step 3, iris strong sprout is transferred in grown cultures liquid, cultivated 2-3 days under natural environment, obtain being used to transplant
Phalaenopsis plants；
The formula of the grown cultures liquid is as follows, and every liter of grown cultures liquid is made up of following components：Compound a great number of elements 1g,
Compound micro substance 1.3-1.5g, inositol 100mg, indolebutyric acid 0.65-0.75mg, peptone 2-3g, activated carbon 1-2g, lemon
Sour 40-50mg, ammonium nitrate 750mg, potassium nitrate 170-180mg, a great number of elements auxiliary agent 6mg, mashed potatoes 45g, banana puree 45g, sugarcane
Sugared 15-20g, agar powder 5g, surplus are water, and the pH of grown cultures liquid is 6.0.
It is preferred that, in above-mentioned iris tissue culture method, the coconut leaching in every liter of Multiplying culture liquid also containing 3g
Juice；Coconut leaching juice in every liter of grown cultures liquid also containing 10g；
Every liter of coconut leaching juice is prepared in accordance with the following methods：Coconut is removed the peel, goes after juice to leave the coconut meat of white, then
Coconut meat is shredded, coconut meat fragment is obtained, 1L water is added into 100g coconut meat fragments, 20min is boiled, is filtered, filtrate, moisturizing is collected
To 1L, coconut leaching juice is obtained.
It is preferred that, in above-mentioned iris tissue culture method, the compound a great number of elements uses a great number of elements mixing
Thing spends one kind in precious No. 1；
The a great number of elements mixture is according to 1 by potassium dihydrogen phosphate, ammonium nitrate, potassium chloride:1:1.4 molar ratio is mixed
Conjunction is formed.
It is preferred that, in above-mentioned iris tissue culture method, the compound micro substance be micro substance mixture or
Kind one kind deposited in multivitamin and minerals tablet (29)；
The micro substance mixture is vitamin A, vitamin D, vitamin E, vitamin B1, vitamin B2, vitamin
B6, vitamin C, vitamin B12, vitamin K1, biotin, folic acid, nicotinic acid, pantothenic acid, calcium chloride, manganese sulfate monohydrate, seven
Water zinc sulphate, Sodium Molybdate Dihydrate, cupric sulfate pentahydrate, CoCL2 6H2O, ferrous sulfate heptahydrate are according to 5:0.4:0.03:1.5:1.7:
2:60:1:0.006:0.025:0.4:20:10:162:2.5:15:5:2:2:1 mass ratio is mixed.
It is preferred that, in above-mentioned iris tissue culture method, a great number of elements auxiliary agent is that agent mixture or China are more
One kind in many No. 1 20-20-20；
The agent mixture is according to 1 by potassium dihydrogen phosphate, ammonium nitrate:1 molar ratio is mixed.
Present invention also offers a kind of iris hyperblastosis culture obtained according to above-mentioned iris tissue culture method
Base, weighs all substances in addition to water according to the formula of the Multiplying culture liquid, fully mixes, and obtains the increasing of iris tissue
Grow culture medium.
Present invention also offers a kind of iris strong seedling culture base obtained according to above-mentioned iris tissue culture method, press
All substances in addition to water are weighed according to the formula of the strong seedling culture liquid, fully mixes, obtains iris strong seedling culture base.
Present invention also offers a kind of iris growth medium obtained according to above-mentioned iris tissue culture method, press
All substances in addition to water are weighed according to the formula of the grown cultures liquid, fully mixes, obtains iris growth medium.
Compared with prior art, method of the invention has the advantages that：
(1) using banana skin and banana pulp as the loss ratio for preparing one of material, reducing banana skin of culture medium, and
Other components are with the addition of on the basis of this, it is ensured that iris tissue germination percentage and cauline leaf differentiation rate are improved, and growth cycle shortens；
Coconut leaching juice is added, while using agricultural wastes, supplemented with trace element and mineral matter, pure chemistry is reduced
The use of material, reduces production cost.
(2) when through boiling the banana puree of processing as culture sill, seedling grow up to the time shorten 12 days, contamination rate be
0, the time that strong sprout grows up to shortens 8 days.Illustrate that influence of the different Feedstock treating modes to iris tissue growth is larger, should
It is advisable when using the banana puree for boiling processing.
(3) Multiplying culture liquid of the invention causes the germination percentage of iris pedicel axillary buds to improve at least 6%, and seedling grows up to
Shorten at least 10 days, improve germination percentage, shorten seedling and grow up to the time；Utilize the butterfly of the strong seedling culture liquid culture of the present invention
Phalaenopsis mesophyll is full, stem branch is sturdy, and more preferably, seedling percent is 100% to upgrowth situation, and strong sprout grows up to the time and shortens at least 14
My god, had a good application prospect in terms of iris tissue cultures and economic value.
Embodiment
With reference to specific embodiment, the present invention is described in detail, but should not be construed as the limitation of the present invention.Below
The experimental method of unreceipted actual conditions in embodiment, is carried out according to the conventional method and condition of this area.
Embodiment 1
A kind of iris tissue culture method, comprises the following steps：
Step 1, the pedicel axillary buds of iris are carried out after routine disinfection, conventional method otch is carried out to pedicel axillary buds, so
After be seeded in Multiplying culture liquid, cultivate 18 days, temperature be 20 DEG C, intensity of illumination 2000Lux, obtain iris seedling；
The formula of the Multiplying culture liquid is as follows, and every liter of proliferated culture medium is made up of following components：Hua Bao companies of the U.S.
HYPONeX's spends precious No. 1 2g, the kind of Wyeth pharmaceuticals to deposit multivitamin and minerals tablet (29) 1.4g, inositol 125mg, 6- benzyl
Purine (BA) 2.5mg, 2,4 dichloro benzene acetic acid (2,4-D) 2.5mg, indolebutyric acid (IBA) 0.5mg, mashed potatoes 100g, sucrose
20g, agar powder 5g, surplus are water, and the pH of Multiplying culture liquid is 5.8；
Step 2, iris seedling is transferred in strong seedling culture liquid, cultivated 24 days, temperature is 20 DEG C, intensity of illumination
2000Lux, obtains iris strong sprout；
The formula of the strong seedling culture liquid is as follows, and every liter of strong seedling culture liquid is made up of following components：Hua Bao companies of the U.S.
HYPONeX's spends precious No. 1 1g, the kind of Wyeth pharmaceuticals to deposit multivitamin and minerals tablet (29) 1.4g, inositol 100mg, peptone
3g, activated carbon 1.5g, citric acid 40mg, ammonium nitrate 650mg, potassium nitrate 175mg, mashed potatoes 40g, banana puree 40g, sucrose 20g,
Agar powder 5g, surplus are water, and the pH of strong seedling culture liquid is 6.0；
Step 3, iris strong sprout is transferred in grown cultures liquid, cultivates 3 days, obtained for transplanting under natural environment
Phalaenopsis plants；
The formula of the grown cultures liquid is as follows, and every liter of grown cultures liquid is made up of following components：Hua Bao companies of the U.S.
HYPONeX's spends precious No. 1 1g, the kind of Wyeth pharmaceuticals to deposit multivitamin and minerals tablet (29) 1.4g, inositol 100mg, indoles fourth
Acid (IBA) 0.70mg, peptone 2.5g, activated carbon 1.5g, citric acid 45mg, ammonium nitrate 750mg, potassium nitrate 175mg, Shi Ke get
Magnificent the more No. 1 20-20-206mg, mashed potatoes 45g, banana puree 45g, sucrose 20g, agar of gardening fertilizer (Wuhan) Co., Ltd
Powder 5g, surplus are water, and the pH of grown cultures liquid is 6.0.
Embodiment 2
A kind of iris tissue culture method, comprises the following steps：
Step 1, after the pedicel axillary buds of iris are carried out disinfection, otch is carried out to pedicel axillary buds, propagation is then seeded to
In nutrient solution, cultivate 12 days, temperature is 25 DEG C, intensity of illumination 1500Lux obtains iris seedling；
The formula of the Multiplying culture liquid is as follows, and every liter of Multiplying culture liquid is made up of following components：Compound a great number of elements 2g,
Compound micro substance 1.3g, inositol 120mg, 6- benzyl purine (BA) 2mg, 2,4 dichloro benzene acetic acid (2,4-D) 2mg, indoles fourth
Acid (IBA) 0.6mg, mashed potatoes 100g, sucrose 25g, agar powder 5g, 3g coconut soak juice, surplus for water, and Multiplying culture liquid
PH is 5.8；
Step 2, iris seedling is transferred in strong seedling culture liquid, cultivated 22 days, temperature is 20 DEG C, intensity of illumination
2000Lux, obtains iris strong sprout；
The formula of the strong seedling culture liquid is as follows, and every liter of strong seedling culture liquid is made up of following components：Compound a great number of elements 1g,
Compound micro substance 1.5g, inositol 100mg, peptone 2.5g, activated carbon 1g, citric acid 35mg, ammonium nitrate 620mg, potassium nitrate
180mg, mashed potatoes 40g, banana puree 45g, sucrose 25g, agar powder 5g, surplus are water, and the pH of strong seedling culture liquid is 6.0；
Step 3, iris strong sprout is transferred in grown cultures liquid, cultivates 2 days, obtained for transplanting under natural environment
Phalaenopsis plants；
The formula of the grown cultures liquid is as follows, and every liter of grown cultures liquid is made up of following components：Compound a great number of elements 1g,
Compound micro substance 1.3g, inositol 100mg, indolebutyric acid (IBA) 0.65mg, peptone 2g, activated carbon 1g, citric acid 50mg,
Ammonium nitrate 750mg, potassium nitrate 170mg, a great number of elements auxiliary agent 6mg, mashed potatoes 45g, banana puree 45g, sucrose 15g, agar powder 5g,
10g coconut leaching juice, surplus are water, and the pH of grown cultures liquid is 6.0.
Embodiment 3
A kind of iris tissue culture method, comprises the following steps：
Step 1, after the pedicel axillary buds of iris are carried out disinfection, otch is carried out to pedicel axillary buds, propagation is then seeded to
In nutrient solution, cultivate 15 days, temperature is 25 DEG C, intensity of illumination 1500Lux obtains iris seedling；
The formula of the Multiplying culture liquid is as follows, and every liter of Multiplying culture liquid is made up of following components：Compound a great number of elements 2g,
Compound micro substance 1.5g, inositol 130mg, 6- benzyl purine (BA) 3mg, 2,4 dichloro benzene acetic acid (2,4-D) 3mg, indoles fourth
Acid (IBA) 0.4mg, mashed potatoes 100g, sucrose 22g, agar powder 5g, surplus are water, and the pH of Multiplying culture liquid is 5.8；
Step 2, iris seedling is transferred in strong seedling culture liquid, cultivated 24 days, temperature is 20 DEG C, intensity of illumination
2000Lux, obtains iris strong sprout；
The formula of the strong seedling culture liquid is as follows, and every liter of strong seedling culture liquid is made up of following components：Compound a great number of elements 1g,
Compound micro substance 1.3g, inositol 100mg, peptone 2g, activated carbon 2g, citric acid 45mg, ammonium nitrate 660mg, potassium nitrate
170mg, mashed potatoes 40g, banana puree 35g, sucrose 22g, agar powder 5g, surplus are water, and the pH of strong seedling culture liquid is 6.0；
Step 3, iris strong sprout is transferred in grown cultures liquid, cultivates 3 days, obtained for transplanting under natural environment
Phalaenopsis plants；
The formula of the grown cultures liquid is as follows, and every liter of grown cultures liquid is made up of following components：Compound a great number of elements 1g,
Compound micro substance 1.5g, inositol 100mg, indolebutyric acid (IBA) 0.75mg, peptone 3g, activated carbon 2g, citric acid 40mg,
Ammonium nitrate 750mg, potassium nitrate 180mg, a great number of elements auxiliary agent 6mg, mashed potatoes 45g, banana puree 45g, sucrose 18g, agar powder 5g,
Surplus is water, and the pH of grown cultures liquid is 6.0.
Embodiment 4
A kind of iris tissue culture method, comprises the following steps：
Step 1, after the pedicel axillary buds of iris are carried out disinfection, otch is carried out to pedicel axillary buds, propagation is then seeded to
In nutrient solution, cultivate 13 days, temperature is 25 DEG C, intensity of illumination 1500Lux obtains iris seedling；
The formula of the Multiplying culture liquid is as follows, and every liter of Multiplying culture liquid is made up of following components：Compound a great number of elements 2g,
Compound micro substance 1.33g, inositol 128mg, 6- benzyl purine (BA) 2.2mg, 2,4 dichloro benzene acetic acid (2,4-D) 2.2mg, Yin
Diindyl butyric acid (IBA) 0.55mg, mashed potatoes 100g, sucrose 25g, agar powder 5g, 3g coconut leaching juice, surplus are water, and breed training
The pH of nutrient solution is 5.8；
Step 2, iris seedling is transferred in strong seedling culture liquid, cultivated 23 days, temperature is 20 DEG C, intensity of illumination
2000Lux, obtains iris strong sprout；
The formula of the strong seedling culture liquid is as follows, and every liter of strong seedling culture liquid is made up of following components：Compound a great number of elements 1g,
Compound micro substance 1.33g, inositol 100mg, peptone 2.6g, activated carbon 1.6g, citric acid 42mg, ammonium nitrate 650mg, nitric acid
Potassium 176mg, mashed potatoes 40g, banana puree 38g, sucrose 23g, agar powder 5g, surplus are water, and the pH of strong seedling culture liquid is 6.0；
Step 3, iris strong sprout is transferred in grown cultures liquid, cultivates 2 days, obtained for transplanting under natural environment
Phalaenopsis plants；
The formula of the grown cultures liquid is as follows, and every liter of grown cultures liquid is made up of following components：Compound a great number of elements 1g,
Compound micro substance 1.45g, inositol 100mg, indolebutyric acid (IBA) 0.72mg, peptone 2.8g, activated carbon 1.2g, citric acid
42mg, ammonium nitrate 750mg, potassium nitrate 174mg, a great number of elements auxiliary agent 6mg, mashed potatoes 45g, banana puree 45g, sucrose 19g, agar
Powder 5g, 10g coconut leaching juice, surplus are water, and the pH of grown cultures liquid is 6.0.
It should be noted that using 0.1mol/L sodium hydroxide or 0.1mol/L salt acid for adjusting pH in above-described embodiment；
Mashed potatoes described in embodiment is to cook fresh potato, then blends into mashed potatoes；The banana puree is by banana skin
10min is steamed together with banana, mud is then blended into, obtains banana puree；
Every liter of coconut leaching juice is prepared in accordance with the following methods：Coconut is removed the peel, goes after juice to leave the coconut meat of white, then
Coconut meat is shredded, coconut meat fragment is obtained, 1L water is added into 100g coconut meat fragments, 20min is boiled, is filtered, filtrate, moisturizing is collected
To 1L, coconut leaching juice is obtained.
It should be noted that in above-described embodiment, the compound a great number of elements uses a great number of elements mixture, described
A great number of elements mixture is according to 1 by potassium dihydrogen phosphate, ammonium nitrate, potassium chloride:1:1.4 molar ratio is mixed；
The compound micro substance is micro substance mixture, the micro substance mixture be vitamin A, vitamin D,
Vitamin E, vitamin B1, vitamin B2, vitamin B6, vitamin C, vitamin B12, vitamin K1, biotin, folic acid, dimension
Raw element PP, pantothenic acid, calcium chloride, manganese sulfate monohydrate, white vitriol, Sodium Molybdate Dihydrate, cupric sulfate pentahydrate, CoCL2 6H2O, seven
Aqueous ferrous sulfate is according to 5:0.4:0.03:1.5:1.7:2:60:1:0.006:0.025:0.4:20:10:162:2.5:15:5:2:
2:1 mass ratio is mixed；
The a great number of elements auxiliary agent be agent mixture, the agent mixture be by potassium dihydrogen phosphate, ammonium nitrate according to
1:1 molar ratio is mixed.
For the ease of preserving, we claim the Multiplying culture liquid in above-described embodiment according to the formula of the Multiplying culture liquid
All substances in addition to water are taken, fully mixes, obtains iris hyperblastosis culture medium, room temperature preservation is standby；According to institute
The formula for stating strong seedling culture liquid weighs all substances in addition to water, fully mixes, obtains iris strong seedling culture base, room temperature
Preservation, it is standby；Formula according to the grown cultures liquid weighs all substances in addition to water, fully mixes, obtains butterfly
Blue growth medium, room temperature preservation is standby.
First, iris growth result is verified
Design spends No. 1 300 groups of treasured, spends precious No. 1 1000 groups, 1 group of embodiment, four groups of embodiment 2 group, training method difference
It is as follows：
Spend precious No. 1 300 groups：Iris pedicel axillary buds are inoculated in and spent in 300 times of liquid of precious No. 1 dilution, 20 DEG C,
Cultivated under the conditions of 2000Lux intensities of illumination, until obtaining iris seedling, observation seedling grows up to time and germination percentage, Ran Houji
Cultivated under the conditions of continuous 20 DEG C, 2000Lux intensities of illumination, until obtaining iris strong sprout, observation strong sprout grows up to time, cauline leaf differentiation
Rate.
Spend precious No. 1 1000 groups：Iris pedicel axillary buds are inoculated in and spent in 1000 times of liquid of precious No. 1 dilution, 20 DEG C,
Cultivated under the conditions of 2000Lux intensities of illumination, until obtaining iris seedling, observation seedling grows up to time and germination percentage, Ran Houji
Cultivated under the conditions of continuous 20 DEG C, 2000Lux intensities of illumination, until obtaining iris strong sprout, observation strong sprout grows up to time, cauline leaf differentiation
Rate.
1 group of embodiment：According to the cultural method described in embodiment 1, observation seedling grows up to time and germination percentage, observes strong sprout
Grow up to time, cauline leaf differentiation rate.
2 groups of embodiment：According to the cultural method described in embodiment 2, observation seedling grows up to time and germination percentage, observes strong sprout
Grow up to time, cauline leaf differentiation rate.
As a result it is as follows：
(1) it is 90% to spend 300 times of liquid germination percentages of precious No. 1 dilution, and the time that seedling grows up to is 28 days；Spend precious No. 1 dilution
1000 times of liquid germination percentages are 80%, and the time that seedling grows up to is 35 days；The Multiplying culture liquid germination percentage of embodiment 1 is 96%,
The time that seedling grows up to is 18 days；The Multiplying culture liquid germination percentage of embodiment 2 is 98%, and the time that seedling grows up to is 12 days.Say
Bright Multiplying culture liquid of the invention make it that the germination percentage of iris pedicel axillary buds improves at least 6%, seedling grow up to shorten to
It is few 10 days, germination percentage is improved, seedling is shortened and grows up to the time.
(2) it is 98% to spend 300 times of liquid cauline leaf differentiation rates of precious No. 1 dilution, and the time that strong sprout grows up to is 38 days；Spend precious No. 1
1000 times of liquid cauline leaf differentiation rates of dilution are 98%, and the time that strong sprout grows up to is 40 days；The Multiplying culture liquid cauline leaf point of embodiment 1
Rate is 100%, and the time that strong sprout grows up to is 24 days；The Multiplying culture liquid cauline leaf differentiation rate of embodiment 2 is 100%, and strong sprout is long
Into time be 22 days.
Compared with spending No. 1 300 groups of treasured, spending No. 1 1000 groups of treasured, the iris leaf of the strong seedling culture liquid culture of the present invention is utilized
Meat is full, stem branch is sturdy, and more preferably, seedling percent is 100% to upgrowth situation, and strong sprout grows up to the time and shortened at least 14 days.
We further demonstrate the nutritive validity of the banana puree of distinct methods preparation, and control group is directly by banana pulp
Mud is blended into together with banana skin, the banana puree of control group is obtained, then according to the method culture iris bennet of embodiment 2
Axillary bud, records observation seedling and grows up to the time, strong sprout grows up to time, cauline leaf differentiation rate, as a result shown, the time that seedling grows up to respectively
For 24 days, contamination rate 2%, the time that strong sprout grows up to was 30 days；
Compared with the data of 2 groups of above-described embodiment, during through boiling the banana puree of processing as culture sill, seedling grows up to
Time shorten 12 days, contamination rate be 0, the time that strong sprout grows up to shortens 8 days.Illustrate different Feedstock treating modes to butterfly
The influence of blue tissue growth is larger, should be advisable using the banana puree for boiling processing.
Those skilled in the art can to the present invention carry out it is various change and modification without departing from the present invention spirit and
Scope, if these modifications and variations of the present invention belong within the scope of the claims in the present invention and its equivalent technologies, then originally
Invention is also intended to comprising including these changes and modification.
Claims (8)
1. a kind of iris tissue culture method, it is characterised in that comprise the following steps：
Step 1, after the pedicel axillary buds of iris are carried out disinfection, otch is carried out to pedicel axillary buds, Multiplying culture is then seeded to
In liquid, cultivate 12-18 days, temperature is 20-25 DEG C, intensity of illumination 1500-2000Lux obtains iris seedling；
The formula of the Multiplying culture liquid is as follows, and every liter of Multiplying culture liquid includes following components：It is compound a great number of elements 2g, compound micro-
Quantity of material 1.3-1.5g, inositol 120-130mg, 6- benzyl purine 2-3mg, 2,4 dichloro benzene acetic acid 2-3mg, indolebutyric acid 0.4-
0.6mg, mashed potatoes 100g, sucrose 20-25g, agar powder 5g, surplus are water, and the pH of Multiplying culture liquid is 5.8；
Step 2, iris seedling is transferred in strong seedling culture liquid, cultivates 22-24 days, obtain iris strong sprout；
The formula of the strong seedling culture liquid is as follows, and every liter of strong seedling culture liquid is made up of following components：It is combined a great number of elements 1g, is combined
Micro substance 1.3-1.5g, inositol 100mg, peptone 2-3g, activated carbon 1-2g, citric acid 35-45mg, ammonium nitrate 620-
660mg, potassium nitrate 170-180mg, mashed potatoes 40g, banana puree 35-45g, sucrose 20-25g, agar powder 5g, surplus are water, and
The pH of strong seedling culture liquid is 6.0；
Step 3, iris strong sprout is transferred in grown cultures liquid, is cultivated 2-3 days under natural environment, obtain the butterfly for transplanting
Phalaenopsis plant；
The formula of the grown cultures liquid is as follows, and every liter of grown cultures liquid includes following components：It is compound a great number of elements 1g, compound micro-
Quantity of material 1.3-1.5g, inositol 100mg, indolebutyric acid 0.65-0.75mg, peptone 2-3g, activated carbon 1-2g, citric acid 40-
50mg, ammonium nitrate 750mg, potassium nitrate 170-180mg, a great number of elements auxiliary agent 6mg, mashed potatoes 45g, banana puree 45g, sucrose 15-
20g, agar powder 5g, surplus are water, and the pH of grown cultures liquid is 6.0.
2. iris tissue culture method according to claim 1, it is characterised in that in every liter of Multiplying culture liquid also
Coconut leaching juice containing 3g；Coconut leaching juice in every liter of grown cultures liquid also containing 10g；
Every liter of coconut leaching juice is prepared in accordance with the following methods：Coconut is removed the peel, goes after juice to leave the coconut meat of white, then by coconut palm
Meat is shredded, and obtains coconut meat fragment, and 1L water is added into 100g coconut meat fragments, 20min is boiled, and is filtered, and collects filtrate, moisturizing is extremely
1L, obtains coconut leaching juice.
3. iris tissue culture method according to claim 1 or 2, it is characterised in that the compound a great number of elements is adopted
It is a great number of elements mixture or spends one kind in precious No. 1；
The a great number of elements mixture is according to 1 by potassium dihydrogen phosphate, ammonium nitrate, potassium chloride:1:1.4 molar ratio mixing and
Into.
4. iris tissue culture method according to claim 1 or 2, it is characterised in that the compound micro substance is
Micro substance mixture or kind one kind deposited in multivitamin and minerals tablet (29)；
The micro substance mixture is vitamin A, vitamin D, vitamin E, vitamin B1, vitamin B2, vitamin B6, dimension
Raw element C, vitamin B12, vitamin K1, biotin, folic acid, nicotinic acid, pantothenic acid, calcium chloride, manganese sulfate monohydrate, seven water sulfuric acid
Zinc, Sodium Molybdate Dihydrate, cupric sulfate pentahydrate, CoCL2 6H2O, ferrous sulfate heptahydrate are according to 5:0.4:0.03:1.5:1.7:2:60:
1:0.006:0.025:0.4:20:10:162:2.5:15:5:2:2:1 mass ratio is mixed.
5. iris tissue culture method according to claim 1 or 2, it is characterised in that a great number of elements auxiliary agent is
One kind in the more No. 1 20-20-20 of agent mixture or China；
The agent mixture is according to 1 by potassium dihydrogen phosphate, ammonium nitrate:1 molar ratio is mixed.
6. the iris hyperblastosis culture medium that a kind of iris tissue culture method according to claim 1 is obtained, it is special
Levy and be, weigh all substances in addition to water according to the formula of the Multiplying culture liquid, fully mix, obtain iris group
Knit proliferated culture medium.
7. a kind of iris strong seedling culture base that iris tissue culture method according to claim 1 is obtained, its feature exists
In weighing all substances in addition to water according to the formula of the strong seedling culture liquid, fully mix, obtain the training of iris strong sprout
Support base.
8. a kind of iris growth medium that iris tissue culture method according to claim 1 is obtained, its feature exists
In weighing all substances in addition to water according to the formula of the grown cultures liquid, fully mix, obtain iris growth training
Support base.
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| CN111631137A (en) * | 2020-07-14 | 2020-09-08 | 郑州市农林科学研究所 | Culture medium and culture method for tissue culture of Sorbus sophorae |
| CN115281081A (en) * | 2021-11-22 | 2022-11-04 | 湘西土家族苗族自治州农业科学研究院 | Breeding method of miniature test tube detoxified seed ginger |
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