CN109804924A - A kind of method of iris tissue culture expanding propagation - Google Patents
A kind of method of iris tissue culture expanding propagation Download PDFInfo
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- CN109804924A CN109804924A CN201711254441.4A CN201711254441A CN109804924A CN 109804924 A CN109804924 A CN 109804924A CN 201711254441 A CN201711254441 A CN 201711254441A CN 109804924 A CN109804924 A CN 109804924A
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Abstract
The present invention relates to a kind of industrial breeding technique fields, more particularly, to a kind of method of iris tissue culture expanding propagation.The technical issues of restriction iris quicker factory nursery the such as mainly expansion of solution iris is numerous slowly for it, the period of taking root is long, program of taking root is complicated and production cost is higher.Method of the invention is then tender shoots to be induced to sprout first to the selection of explant and disinfection, then be Multiplying culture and culture of rootage, finally carry out rooting culture.In the culture medium that wherein induction tender shoots sprouts, nutrient media components are as follows: the MS minimal medium+6-BA1-2mg/L+IBA0.3-0.5mg/L+ white sugar 25-35mg/L+ agar 5g/L of improvement, pH is adjusted to 5.8, and it is to replace original potassium nitrate and calcium chloride with nitric hydrate calcium and potassium sulfate that the WPM of improvement is cultivated substantially；Rooting culture carries out root dipping processing again.
Description
Technical field
The present invention relates to a kind of industrial breeding technique fields, more particularly, to a kind of iris tissue culture expanding propagation and outside sprout-cultivating-bottle
Method.
Background technique
Iris also known as: blue butterfly, The purple butterfly, flat bamboo etc., belong to Asparagales, Iridaceae herbaceos perennial.With
The raising of people's aesthetic conceptions, common greening have been unable to meet the demand of people, and people thirst for looking for novelty, asking different, and eye is made us in serious hope
Previous bright garden landscape.Vegetable material abundant is the element that garden landscape is formed, and tall shrub stereoscopic configurations are Landscape arrangements
Principle is also the demand of an ecosystem bio-diversity.As ground cover plant material, surely belong in addition to lawn with type and
In a great variety, resistance is strong, the Perennial Flowers for the characteristic that cultivation management is extensive and plantation in 1 year is ornamental for many years.Come relative to lawn
It says, Perennial Flowers can save a large amount of water resource, be optimal green material application for the area and region of shortage of water resources.
Have outstanding performance in Perennial Flowers is Jris.Iris is many kinds of, and flower-shape is different, and characteristic is had nothing in common with each other.Since iris type is more,
So the application in gardens is quite extensive.Dutch iris Vase time length is suitble to do cut-flower；Short raw iris be suitble to rock garden and
Presbyopic glasses fringing；It plants the suitable waterside of humidogene；The raw suitable slope greening of drought；It is glad for people that iris can also build up individual Botanical Garden
Reward.
Iris is wide in variety, and adaptability is extensive, but the kind being applied in China's afforestation is limited, the landscape list of formation
Adjust dull, and modes of reproduction is single, is not able to satisfy market and the demand of people.
Currently, the iris of China's cultivation is usually based on proliferation in production.In recent years, pass through scientific research personnel's
It is continually striving to, and the further investigation to tissue culture technique, having explored one kind for various regions being capable of relatively simple quick kite
Tail propagation method, tissue cultures factorial seedling-culturing method are widely used to iris seedling breeding.It the development of this technology and answers
With providing a large amount of nontoxic good seedlings to iris production, be increasingly becoming the direction of iris high quality seedling production from now on.But
There are many problems in iris tissue culture procedures, such as: rooting rate is low, the period of taking root is long, program of taking root is complicated and takes root
Higher cost etc. cannot achieve iris quicker factory nursery.
Summary of the invention
The present invention is to provide a kind of method of iris tissue culture expanding propagation, mainly solves iris proliferative speed slowly and takes root
Higher cost etc. restricts the technical issues of iris quicker factory nursery.
Above-mentioned technical problem of the invention is mainly to be addressed by following technical proposals:
A. the selection and disinfection of explant: selecting the no disease and pests harm plant of robust growth, and cutting tender shoots, (root segment is outstanding white
Color part, the partial volume cut are greater than tender shoots), surface is cleared up with hairbrush, rinses under subsequent flowing water, is used on superclean bench
Alcohol carries out disinfection, and then uses aseptic water washing 2-3 times, is transferred in mercuric chloride solution and carries out disinfection, then with aseptic water washing 3-4
It is secondary, it is rear spare.
B. induction tender shoots sprouts: tender shoots obtained in the previous step being cut off redundance in superclean bench, is inoculated into and lures
In the culture medium for leading tender shoots sprouting, nutrient media components are as follows: 1/2MS minimal medium+6-BA
3-4mg/L+IBA0.2-0.4mg/L+ white sugar 25-35mg/L+ agar 5g/L, pH are adjusted to 5.8；
C. Multiplying culture: by the tender shoots of axenic germination obtained in the previous step be transferred to subculture in proliferated culture medium expand it is numerous,
Proliferation culture medium formula component are as follows: the MS minimal medium+6-BA1-2mg/L+IBA0.3-0.5mg/L+ white sugar 25- of improvement
35mg/L+ agar 5g/L, pH are adjusted to 5.8；
D. culture of rootage: the tender shoots that proliferation is obtained carries out budding, is transferred to culture of rootage in root media, culture of rootage
Based formulas component are as follows: MS minimal medium+paclobutrazol 0.6-0.8mg/L+ white sugar 25-35mg/L+ agar 5g/L, pH is adjusted to 5.8
E. rooting culture: by 8-10cm stalwartness aseptic seedling obtained in the previous step, root long opens bottle cap, is covered with wet towel logical
During which hardening 2-3 days at wind pay attention to that towel is kept to be in moisture state, seedbed and matrix are handled and disappeared before transplanting domestication
Poison, matrix use turfy soil and vermiculite, and ratio range is between 1: 1 and 2: 1, after disinfection, the thickness 4-6cm in seedbed；It will be through
Cross ventilation hardening healthy and strong iris tissue-cultured seedling, cuttage into above-mentioned seedbed, every section of need of cuttage process dip root-growing agent, cuttage
Seedbed overlay film after completion, relative humidity control is in 60-80% in canopy, and temperature is controlled at 23-28 DEG C, once every spray in 2-3 days
Hereafter water sprayed primary flower the more foliar fertilizer every 10 days, finally obtain iris and take root finished product seedling.
The present invention is proliferated the MS minimal medium culture medium using improvement, is replaced with nitric hydrate calcium and potassium sulfate original
Potassium nitrate and calcium chloride, so that proliferation times of iris during Multiplying culture be made to improve 2-3 times, using after taking root in bottle
It carries out root dipping processing technique, hestening rooting again outside, thus shortens the period of industrial seedling rearing, reduce tissue culture cost, together
When improve transplanting hardening survival rate, its multiplication rate is higher by much compared with iris tissue-cultured seedling, to reduce breeding
Cost.
Preferably, MS culture medium is the culture medium of improvement in the step c
Preferably, the carbendazim that the disinfection of the step d mesostroma need to be 1000 times with concentration sterilizes 24 hours.
Preferably, the root-growing agent is 1.0-1.5mg/L NAA and 0.5-0.8mg/L IBA mixed liquor, when root dipping
Between be 5-10s.
Specific embodiment
Below by embodiment, the technical solutions of the present invention will be further described.
Embodiment 1: a kind of method of iris (India head) tissue culture expanding propagation of this example the steps include:
A. the selection and disinfection of explant: selecting the no disease and pests harm plant of robust growth, and cutting tender shoots, (root segment is outstanding white
Color part, the partial volume cut are greater than tender shoots), surface is cleared up with hairbrush, rinses under subsequent flowing water, is used on superclean bench
Alcohol carries out disinfection, and then uses aseptic water washing 2-3 times, is transferred in mercuric chloride solution and carries out disinfection, then with aseptic water washing 3-4
It is secondary, it is rear spare.
B. induction tender shoots sprouts: tender shoots obtained in the previous step being cut off redundance in superclean bench, is inoculated into and lures
In the culture medium for leading tender shoots sprouting, nutrient media components are as follows: 1/2MS minimal medium+6-BA
3-4mg/L+IBA0.2-0.4mg/L+ white sugar 25-35mg/L+ agar 5g/L, pH are adjusted to 5.8；
C. Multiplying culture: by the tender shoots of axenic germination obtained in the previous step be transferred to subculture in proliferated culture medium expand it is numerous,
Proliferation culture medium formula component are as follows: the MS minimal medium+6-BA1-2mg/L+IBA0.3-0.5mg/L+ white sugar 25- of improvement
35mg/L+ agar 5g/L, pH are adjusted to 5.8；
D. culture of rootage: the tender shoots that proliferation is obtained carries out budding, is transferred to culture of rootage in root media, culture of rootage
Based formulas component are as follows: MS minimal medium+paclobutrazol 0.6-0.8mg/L+ white sugar 25-35mg/L+ agar 5g/L, pH is adjusted to 5.8.
E. rooting culture: by 8-10cm stalwartness aseptic seedling obtained in the previous step, root long opens bottle cap, is covered with wet towel logical
During which hardening 2-3 days at wind pay attention to that towel is kept to be in moisture state, seedbed and matrix are handled and disappeared before transplanting domestication
Poison, matrix use turfy soil and vermiculite, and ratio range is between 1: 1 and 2: 1, after disinfection, the thickness 4-6cm in seedbed；It will be through
The healthy and strong iris tissue-cultured seedling of ventilation hardening is crossed, cuttage is into above-mentioned seedbed, and every section of need of cuttage process dip root-growing agent, and cuttage is complete
At seedbed overlay film later, in 60-80%, temperature is controlled at 23-28 DEG C for relative humidity control in canopy, every the water of spray in 2-3 days,
Hereafter primary flower the more foliar fertilizer was sprayed every 10 days, is finally obtained iris and is taken root finished product seedling.
Its condition of culture of establishing of above-mentioned iris sterile system is: 25 ± 2 DEG C of temperature, intensity of illumination 2500-3000lx, light
According to time 14-16 hour.
Embodiment 2: a kind of method of iris (sky blue) tissue culture expanding propagation of this example the steps include:
A. the selection and disinfection of explant: selecting the no disease and pests harm plant of robust growth, and cutting tender shoots, (root segment is outstanding white
Color part, the partial volume cut are greater than tender shoots), surface is cleared up with hairbrush, rinses under subsequent flowing water, is used on superclean bench
Alcohol carries out disinfection, and then uses aseptic water washing 2-3 times, is transferred in mercuric chloride solution and carries out disinfection, then with aseptic water washing 3-4
It is secondary, it is rear spare.
B. induction tender shoots sprouts: tender shoots obtained in the previous step being cut off redundance in superclean bench, is inoculated into and lures
In the culture medium for leading tender shoots sprouting, nutrient media components are as follows: 1/2MS minimal medium+6-BA
3-4mg/L+IBA0.2-0.4mg/L+ white sugar 25-35mg/L+ agar 5g/L, pH are adjusted to 5.8；
C. Multiplying culture: by the tender shoots of axenic germination obtained in the previous step be transferred to subculture in proliferated culture medium expand it is numerous,
Proliferation culture medium formula component are as follows: the MS minimal medium+6-BA1-2mg/L+IBA0.3-0.5mg/L+ white sugar 25- of improvement
35mg/L+ agar 5g/L, pH are adjusted to 5.8；
D. culture of rootage: the tender shoots that proliferation is obtained carries out budding, is transferred to culture of rootage in root media, culture of rootage
Based formulas component are as follows: MS minimal medium+paclobutrazol 0.6-0.8mg/L+ white sugar 25-35mg/L+ agar 5g/L, pH is adjusted to 5.8
E. rooting culture: by 8-10cm stalwartness aseptic seedling obtained in the previous step, root long opens bottle cap, is covered with wet towel logical
During which hardening 2-3 days at wind pay attention to that towel is kept to be in moisture state, seedbed and matrix are handled and disappeared before transplanting domestication
Poison, matrix use turfy soil and vermiculite, and ratio range is between 1: 1 and 2: 1, after disinfection, the thickness 4-6cm in seedbed；It will be through
The healthy and strong iris tissue-cultured seedling of ventilation hardening is crossed, cuttage is into above-mentioned seedbed, and every section of need of cuttage process dip root-growing agent, and cuttage is complete
At seedbed overlay film later, in 60-80%, temperature is controlled at 23-28 DEG C for relative humidity control in canopy, every the water of spray in 2-3 days,
Hereafter primary flower the more foliar fertilizer was sprayed every 10 days, is finally obtained iris and is taken root finished product seedling.
Its condition of culture of establishing of above-mentioned iris sterile system is: 25 ± 2 DEG C of temperature, intensity of illumination 2500-3000lx, light
According to time 14-16 hour.
Above is only a specific embodiment of the present invention, but structure feature of the invention is not limited thereto, Ren Heben
Within the field of the present invention, made changes or modifications all cover within the scope of the patent of the present invention the technical staff in field.
Claims (8)
1. a kind of method of iris tissue culture expanding propagation and outside sprout-cultivating-bottle characterized by comprising
Step a. tender shoots carries out disinfection processing；
Step b: it will be inoculated into the culture medium that induction tender shoots sprouts by the tender shoots of step a disinfection, culture；
Step c: it is numerous that the tender shoots of the step b axenic germination obtained be transferred to subculture expansion in proliferated culture medium；
Step d: the step c tender shoots for expanding numerous acquisition is subjected to budding, is transferred to culture of rootage in root media；
Step e: to seedbed, enchanting simultaneously controls humidity for transplanting after the 8-10cm stalwartness aseptic seedling that step d is obtained carries out hardening
60%~80%, temperature is controlled at 23 DEG C -28 DEG C range, every the water of spray in 2-3 days, hereafter sprays primary flower every 10 days
The more foliar fertilizer finally obtains iris and takes root finished product seedling；
The culture medium that the induction tender shoots sprouts are as follows: 1/2MS minimal medium+6-BA 3-4mg/L+IBA 0.2-0.4mg/L+
White sugar 25-35mg/L+ agar 5g/L, pH are adjusted to 5.8；
The proliferated culture medium are as follows: the MS minimal medium+6-BA 1-2mg/L+IBA 0.3-0.5mg/L+ white sugar 25- of improvement
35mg/L+ agar 5g/L, pH are adjusted to 5.8；
The root media are as follows: MS minimal medium+paclobutrazol 0.6-0.8mg/L+ white sugar 25-35mg/L+ agar 5g/L, pH
It is adjusted to 5.8；
The matrix in the seedbed is (1~2) by mass ratio: 1 turfy soil and vermiculite forms；The thickness 4-6cm in the seedbed.
2. the method according to claim 1, wherein the disinfection treatment includes: the disease-free of selection robust growth
Insect pest plant, cuts tender shoots, clears up surface with hairbrush, rinses under subsequent flowing water, carried out disinfection on superclean bench with alcohol,
Then it uses aseptic water washing 2-3 times, is transferred in mercuric chloride solution and carries out disinfection, then with aseptic water washing 3-4 times, it is rear spare.
3. the method according to claim 1, wherein with nitric hydrate calcium in the MS minimal medium of the improvement
Original potassium nitrate and calcium chloride are replaced with potassium sulfate.
4. the method according to claim 1, wherein condition of culture is in step b~d: 25 ± 2 DEG C of temperature, light
According to intensity 2500-3000lx, light application time 14-16 hours.
5. the method according to claim 1, wherein being transplanted after the aseptic seedling after hardening dips in root-growing agent；Institute
Stating root-growing agent includes 1.0-1.5mg/L NAA and 0.5-0.8mg/L IBA, and the root dipping time is 5-10s.
6. described the method according to claim 1, wherein carry out disinfection before the transplanting to seedbed and matrix
Disinfection uses concentration to sterilize 24 hours for 1000 times of carbendazim.
7. being covered with wet towel the method according to claim 1, wherein the hardening is to open bottle cap and divulging information
During which place hardening 2-3 days pays attention to that towel is kept to be in moisture state.
8. the method according to claim 1, wherein the kind of the iris is India head or sky blue.
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| CN201711254441.4A CN109804924A (en) | 2017-11-22 | 2017-11-22 | A kind of method of iris tissue culture expanding propagation |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114403000A (en) * | 2021-12-31 | 2022-04-29 | 湖北懿昌能兴生态农业科技有限公司 | Louisiana iris tissue culture propagation method with high plant regeneration rate |
| CN115250837A (en) * | 2022-07-29 | 2022-11-01 | 蒙草生态环境(集团)股份有限公司 | Method for delaying growth and increasing tillering of bluegrass |
-
2017
- 2017-11-22 CN CN201711254441.4A patent/CN109804924A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114403000A (en) * | 2021-12-31 | 2022-04-29 | 湖北懿昌能兴生态农业科技有限公司 | Louisiana iris tissue culture propagation method with high plant regeneration rate |
| CN115250837A (en) * | 2022-07-29 | 2022-11-01 | 蒙草生态环境(集团)股份有限公司 | Method for delaying growth and increasing tillering of bluegrass |
| CN115250837B (en) * | 2022-07-29 | 2023-07-14 | 蒙草生态环境(集团)股份有限公司 | Method for delaying growth and increasing tillering of meadow bluegrass |
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