CN108056024A - A kind of Herba Diploprora championii sprouting and rooting method - Google Patents
A kind of Herba Diploprora championii sprouting and rooting method Download PDFInfo
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- CN108056024A CN108056024A CN201810142286.5A CN201810142286A CN108056024A CN 108056024 A CN108056024 A CN 108056024A CN 201810142286 A CN201810142286 A CN 201810142286A CN 108056024 A CN108056024 A CN 108056024A
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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 present invention provides a kind of method of Herba Diploprora championii sprouting and rooting,Belong to orchid germ plasm resource regeneration field,Gather Herba Diploprora championii capsule,Aseptically carry out bud induction,It is proliferated and takes root while complete Fiber differentiation,Obtain the Herba Diploprora championii seedling that can be transplanted,The two step seedling methods of orchid,It is long to solve orchid cultivation cycle,It is of high cost that productions problem is waited to solve some precious Cymbidium source resources difficulties in the prior art simultaneously,It is born in the mountainous region woods of 250 1450 meters of height above sea level on trunk or is required on cheuch rock and to environmental condition harsh,Wild resource is few,Appreciation effect is good,Naturally ability is multiplied,Spreading and diffusion is indifferent,Natural renovation is difficult,The problems such as good seed is rare,It is Chinese Second Class Key Protected Plant,With higher gardening ornamental value,It is of great significance for orchid gardens decoration and orchid park decoration.
Description
Technical field
The invention belongs to orchid germ plasm resource regeneration fields, and in particular to a kind of method of Herba Diploprora championii sprouting and rooting.
Background technology
Herba Diploprora championii（Scientific name：Diploprora championii (Lindl.) Hook. f.）：Stem hard texture, it is cylindrical
Or slightly flat cylinder, it is often sagging.Leaf papery, sickle shaped lanceolar or plagioclase are circular, the sheath that base portion tool harbors, edge ripple sometimes
Shape.Raceme is with leaf to life, longer than leaf or short, sagging, 2-5 flower of tool；Flower tool fragrance, slightly meat, carry out, sepal and petal
It is faint yellow；Petal is smaller than sepal；Lip white band rosiness, middle part with lower recess in boat-shaped, without away from, be about 10 millimeters, it is wide
About 4 millimeters, slightly 3 split.Capsule is cylindrical.The month at florescence 2-8, the fruiting period 3-9 months.
It is born in the mountainous region woods of 250-1450 meters of height above sea level on trunk or on cheuch rock.It is distributed in China, Sri Lanka, print
Degree（Deccan Traps）, Sillim, Burma, Thailand, Vietnam.This flower has cultivation, has higher horticultural importance.
The content of the invention
It is an object of the invention to provide a kind of methods of Herba Diploprora championii sprouting and rooting, solve orchid cultivation cycle
It is long, it is of high cost that productions problem is waited to solve some precious Cymbidium source resources difficulties in the prior art simultaneously, it is born in height above sea level 250-
Harshness is required in 1450 meters of mountainous region woods on trunk or on cheuch rock and to environmental condition, wild resource is few, and appreciation effect is good,
Naturally the problems such as ability, Spreading and diffusion are indifferent, and natural renovation is difficult, good seed is rare is multiplied, is national second class protection
Plant has higher gardening ornamental value, is of great significance for orchid gardens decoration and orchid park decoration.
To achieve the above object, the present invention adopts the following technical scheme that：
A kind of Herba Diploprora championii sprouting and rooting method, includes the following steps：
1）Material selection and disinfection：Picking 90% ripe pod detergent rinsed clean, then be placed in flowing water and rinse 20 points then
Clock；Seed disinfection is handled：Pod is cleaned, puts and 15min is impregnated in saturation bleaching powder supernatant, and pod is scrubbed with banister brush
Outside fruit, after rinsing well, drop rushes 0.5h under tap water, and distilled water dashes 2-3 times, puts in superclean bench and is disappeared with 75% alcohol
It removes alcohol after malicious 30s, adds in 0.1% mercuric chloride processing 5-8min, mercury solution is poured into useless mercury bottle, sterile water rushes 3-4 after, with disappearing
After malicious filter paper blots surface moisture, for use；
2）Bud inducement cultivation：Take it is full it is sterile-processed after pod, clamp pod with gun-shaped forceps, scalpel is vertical along pod
Direction is splitted, and is taken out seed in pod, is inoculated in respectively through in high temperature, autoclaved bud inducement cultivation base；Condition of culture：Training
It is 23 ± 2 DEG C to support room temperature, and after no optical culture 69d, 70d light application times set 3h, intensity of illumination 500lx, afterwards 10d illumination
Time 5h, intensity of illumination 500-1000lx, incubation time 80d（It all sprouts for 80d）, the growth plant that is directly induced；
3）Multiplication takes root while completes to cultivate：Will be through above-mentioned Fiber differentiation, acquired well-grown clump bud is according to plant size
Plant division inoculation is carried out, high 0.3-0.5cm, the plantlet of blade 1-2 pieces are inoculated in bud proliferated culture medium, high about 0.5-1.0cm,
The big plant of blade 2-3 pieces is inoculated in the culture medium for completing rooting process, light application time 8h/d, intensity of illumination 1500-
2000lx, Multiplying culture time 35-45d obtain multiplication and take root while complete to grow seedlings；
4）The culture of test tube bottle seedling is completed：When culture of rootage test tube seedling grows supreme 5-6cm, root 3-5 items, leaf 3-5 pieces, the long 1.5- of leaf
The germination of Herba Diploprora championii Seed inducement and quick breeding culture are completed during 2.5cm；
6）It will complete culture of rootage test tube bottle seedling and be placed on natural light lower refining seedling 5-7 days, bottle cap hardening 1-2d is opened, to enhance test tube
Seedling is to the adaptability of outdoor environment；Then taken out from blake bottle, clean the remaining medium for being attached to root system, move into humic
Soil：Water plant mass ratio is 1:In the matrix of 2 mixing, moisturizing shading, temperature is controlled at 15-30 DEG C, and humidity is maintained at 75%-
85%, direct sunlight is avoided,；Plant adaptability gradually enhances after 2-3 weeks, obtains the complete seedling of nature robust growth.
The bud inducement cultivation base is：MS+1.0mg/L spends No. 1+0.5mg/LNAA+1.0g/L peptones+20g/L of treasured
+ 1.0 gL of sucrose+5.8g/L agar powders-1Activated carbon；PH value is 5.4-5.7.
The bud proliferated culture medium：1/2MS+1.0g/L spend precious No. 2+0.5mg/LKT+0.1mg/LNAA+25g/L sugarcanes
Sugar+5.6g/L agar powders+1.50gL-1Activated carbon；PH value is 5.4-5.7.
The culture medium of the rooting process spends No. 2+0.5mg/LKT+0.1mg/LNAA+25g/ of treasured for 1/2MS+1.0g/L
L sucrose+5.6g/L agar powders+1.50gL-1Activated carbon；PH value is 5.4-5.7.
The advantage of the invention is that：
1st, the selection of Establishing material：Maturity is the 90% full pod that do not crack then for picking；2nd, correct disinfecting time and
The processing method of material；3rd, accurate rational medium component is formed；4th, the setting of rational light application time intensity of illumination improves
Germination rate；5th, multiplication takes root while completes to cultivate, and root induction rate is up to 100%, and survival rate is up to more than 95%；Incubation time
Survival rate is high after shortening seedling transplanting, and pest and disease damage is few, reduces great amount of cost, and seedling stalwartness is tall and straight, grows fine, and neat one
It causes.
Description of the drawings
Fig. 1 is the bud plant after Fiber differentiation.
The seedling completed simultaneously after Fig. 2 multiplication culture of rootage.
Specific embodiment
Embodiment 1
1）Material selection and disinfection：Picking 90% ripe pod detergent rinsed clean, then be placed in flowing water and rinse 20 points then
Clock；Seed disinfection is handled：Pod is cleaned, puts and 15min is impregnated in saturation bleaching powder supernatant, and pod is scrubbed with banister brush
Outside fruit, after rinsing well, drop rushes 0.5h under tap water, and distilled water dashes 3 times, puts in superclean bench with 75% alcohol disinfecting
Alcohol is removed after 30s, adds in 0.1% mercuric chloride processing 6min, mercury solution is poured into useless mercury bottle, after sterile water rushes 4 times, uses sterilized filter paper
After blotting surface moisture, for use；
2）Bud inducement cultivation：Take it is full it is sterile-processed after pod, clamp pod with gun-shaped forceps, scalpel is vertical along pod
Direction is splitted, and is taken out seed in pod, is inoculated in respectively through in high temperature, autoclaved bud inducement cultivation base；Condition of culture：Training
It is 23 ± 2 DEG C to support room temperature, and after no optical culture 69d, 70d, light application time sets 3h, and 10d light application times 5h, illumination are strong afterwards
Spend 800lx, incubation time 80d, the growth plant directly induced；
3）Multiplication takes root while completes to cultivate：Will be through above-mentioned Fiber differentiation, acquired well-grown clump bud is according to plant size
Plant division inoculation is carried out, high 0.3-0.5cm, the plantlet of blade 1-2 pieces are inoculated in bud proliferated culture medium, high 0.5-1.0cm, leaf
The big plant of piece 2-3 pieces is inoculated in the culture medium for completing rooting process, light application time 8h/d, intensity of illumination 1800lx, multiplication
Incubation time 40d obtains multiplication and takes root while complete to grow seedlings；
4）The culture of test tube bottle seedling is completed：When culture of rootage test tube seedling grows supreme 6cm, root 4,4, leaf, when leaf long 2cm completes
Herba Diploprora championii Seed inducement germinates and quick breeding culture；
6）It will complete culture of rootage test tube bottle seedling and be placed on natural light lower refining seedling 6 days, bottle cap hardening 2d is opened, to enhance test tube seedling pair
The adaptability of outdoor environment；Then taken out from blake bottle, clean the remaining medium for being attached to root system, move into fertile soil：
Water plant mass ratio is 1:In the matrix of 2 mixing, moisturizing shading, temperature is controlled at 25 DEG C, and humidity is maintained at 80%, avoids sunlight straight
It penetrates,；Plant adaptability gradually enhances after 3 weeks, obtains the complete seedling of nature robust growth.
The bud inducement cultivation base is：MS+1.0mg/L spends No. 1+0.5mg/LNAA+1.0g/L peptones+20g/L of treasured
+ 1.0 gL of sucrose+5.8g/L agar powders-1Activated carbon；PH value is 5.6.
The bud proliferated culture medium：1/2MS+1.0g/L spend precious No. 2+0.5mg/LKT+0.1mg/LNAA+25g/L sugarcanes
Sugar+5.6g/L agar powders+1.50gL-1Activated carbon；PH value is 5.6.
The culture medium of the rooting process spends No. 2+0.5mg/LKT+0.1mg/LNAA+25g/ of treasured for 1/2MS+1.0g/L
L sucrose+5.6g/L agar powders+1.50gL-1Activated carbon；PH value is 5.6.
Survival rate is to 100%.
Embodiment 2
1）Material selection and disinfection：Picking 90% ripe pod detergent rinsed clean, then be placed in flowing water and rinse 20 points then
Clock；Seed disinfection is handled：Pod is cleaned, puts and 15min is impregnated in saturation bleaching powder supernatant, and pod is scrubbed with banister brush
Outside fruit, after rinsing well, drop rushes 0.5h under tap water, and distilled water dashes 2 times, puts in superclean bench with 75% alcohol disinfecting
Alcohol is removed after 30s, adds in 0.1% mercuric chloride processing 5min, mercury solution is poured into useless mercury bottle, after sterile water rushes 3 times, uses sterilized filter paper
After blotting surface moisture, for use；
2）Bud inducement cultivation：Take it is full it is sterile-processed after pod, clamp pod with gun-shaped forceps, scalpel is vertical along pod
Direction is splitted, and is taken out seed in pod, is inoculated in respectively through in high temperature, autoclaved bud inducement cultivation base；Condition of culture：Training
It is 23 ± 2 DEG C to support room temperature, and after no optical culture 69d, 70d light application times set 3h, intensity of illumination 500lx, afterwards 10d illumination
Time 5h, intensity of illumination 500-1000lx, incubation time 80d（It all sprouts for 80d）, the growth plant that is directly induced；
3）Multiplication takes root while completes to cultivate：Will be through above-mentioned Fiber differentiation, acquired well-grown clump bud is according to plant size
Plant division inoculation is carried out, high 0.3-0.5cm, the plantlet of blade 1-2 pieces are inoculated in bud proliferated culture medium, high about 0.5-1.0cm,
The big plant of blade 2-3 pieces is inoculated in the culture medium for completing rooting process, and light application time 8h/d, intensity of illumination 1500lx increase
Incubation time 35-45d is grown, multiplication is obtained and takes root while complete to grow seedlings；
4）The culture of test tube bottle seedling is completed：When culture of rootage test tube seedling grows supreme 5cm, root 3,3, leaf, when leaf long 1.5cm is complete
Into the germination of Herba Diploprora championii Seed inducement and quick breeding culture；
6）It will complete culture of rootage test tube bottle seedling and be placed on natural light lower refining seedling 5 days, bottle cap hardening 1d is opened, to enhance test tube seedling pair
The adaptability of outdoor environment；Then taken out from blake bottle, clean the remaining medium for being attached to root system, move into fertile soil：
Water plant mass ratio is 1:In the matrix of 2 mixing, moisturizing shading, temperature is controlled at 15 DEG C, and humidity is maintained at 75%, avoids sunlight straight
It penetrates；Plant adaptability gradually enhances after 2 weeks, obtains the complete seedling of nature robust growth.
The bud inducement cultivation base is：MS+1.0mg/L spends No. 1+0.5mg/LNAA+1.0g/L peptones+20g/L of treasured
+ 1.0 gL of sucrose+5.8g/L agar powders-1Activated carbon；PH value is 5.4.
The bud proliferated culture medium：1/2MS+1.0g/L spend precious No. 2+0.5mg/LKT+0.1mg/LNAA+25g/L sugarcanes
Sugar+5.6g/L agar powders+1.50gL-1Activated carbon；PH value is 5.4.
The culture medium of the rooting process spends No. 2+0.5mg/LKT+0.1mg/LNAA+25g/ of treasured for 1/2MS+1.0g/L
L sucrose+5.6g/L agar powders+1.50gL-1Activated carbon；PH value is 5.4.
Survival rate is to 99%.
Embodiment 3
1）Material selection and disinfection：Picking 90% ripe pod detergent rinsed clean, then be placed in flowing water and rinse 20 points then
Clock；Seed disinfection is handled：Pod is cleaned, puts and 15min is impregnated in saturation bleaching powder supernatant, and pod is scrubbed with banister brush
Outside fruit, after rinsing well, drop rushes 0.5h under tap water, and distilled water dashes 3 times, puts in superclean bench with 75% alcohol disinfecting
Alcohol is removed after 30s, adds in 0.1% mercuric chloride processing 8min, mercury solution is poured into useless mercury bottle, after sterile water rushes 4 times, uses sterilized filter paper
After blotting surface moisture, for use；
2）Bud inducement cultivation：Take it is full it is sterile-processed after pod, clamp pod with gun-shaped forceps, scalpel is vertical along pod
Direction is splitted, and is taken out seed in pod, is inoculated in respectively through in high temperature, autoclaved bud inducement cultivation base；Condition of culture：Training
It is 23 ± 2 DEG C to support room temperature, and after no optical culture 69d, 70d light application times set 3h, intensity of illumination 500lx, afterwards 10d illumination
Time 5h, intensity of illumination 1000lx, incubation time 80d, the growth plant directly induced；
3）Multiplication takes root while completes to cultivate：Will be through above-mentioned Fiber differentiation, acquired well-grown clump bud is according to plant size
Carry out plant division inoculation, high 0.5cm, the plantlet of 2, blade be inoculated in bud proliferated culture medium, high about 1.0cm, 3, blade
Big plant is inoculated in the culture medium for completing rooting process, light application time 8h/d, intensity of illumination 2000lx, the Multiplying culture time
45d obtains multiplication and takes root while complete to grow seedlings；
4）The culture of test tube bottle seedling is completed：When culture of rootage test tube seedling grows supreme 6cm, root 5,5, leaf, when leaf long 2.5cm is complete
Into the germination of Herba Diploprora championii Seed inducement and quick breeding culture；
6）It will complete culture of rootage test tube bottle seedling and be placed on natural light lower refining seedling 7 days, bottle cap hardening 2d is opened, to enhance test tube seedling pair
The adaptability of outdoor environment；Then taken out from blake bottle, clean the remaining medium for being attached to root system, move into fertile soil：
Water plant mass ratio is 1:In the matrix of 2 mixing, moisturizing shading, temperature is controlled at 30 DEG C, and humidity is maintained at 85%, avoids sunlight straight
It penetrates,；Plant adaptability gradually enhances after 3 weeks, obtains the complete seedling of nature robust growth.
The bud inducement cultivation base is：MS+1.0mg/L spends No. 1+0.5mg/LNAA+1.0g/L peptones+20g/L of treasured
+ 1.0 gL of sucrose+5.8g/L agar powders-1Activated carbon；PH value is 5.7.
The bud proliferated culture medium：1/2MS+1.0g/L spend precious No. 2+0.5mg/LKT+0.1mg/LNAA+25g/L sugarcanes
Sugar+5.6g/L agar powders+1.50gL-1Activated carbon；PH value is 5.7.
The culture medium of the rooting process spends No. 2+0.5mg/LKT+0.1mg/LNAA+25g/ of treasured for 1/2MS+1.0g/L
L sucrose+5.6g/L agar powders+1.50gL-1Activated carbon；PH value is 5.7.
Survival rate is to 99%.
The foregoing is merely presently preferred embodiments of the present invention, all equivalent changes done according to scope of the present invention patent with
Modification should all belong to the covering scope of the present invention.
Claims (4)
- A kind of 1. Herba Diploprora championii sprouting and rooting method, it is characterised in that：Include the following steps：1）Material selection and disinfection：Picking 90% ripe pod detergent rinsed clean, then be placed in flowing water and rinse 20 points then Clock；Seed disinfection is handled：Pod is cleaned, puts and 15min is impregnated in saturation bleaching powder supernatant, and pod is scrubbed with banister brush Outside fruit, after rinsing well, drop rushes 0.5h under tap water, and distilled water dashes 2-3 times, puts in superclean bench and is disappeared with 75% alcohol It removes alcohol after malicious 30s, adds in 0.1% mercuric chloride processing 5-8min, mercury solution is poured into useless mercury bottle, sterile water rushes 3-4 after, with disappearing After malicious filter paper blots surface moisture, for use；2）Bud inducement cultivation：Take it is full it is sterile-processed after pod, clamp pod with gun-shaped forceps, scalpel is vertical along pod Direction is splitted, and is taken out seed in pod, is inoculated in respectively through in high temperature, autoclaved bud inducement cultivation base；Condition of culture：Training It is 23 ± 2 DEG C to support room temperature, and after no optical culture 69d, 70d light application times set 3h, intensity of illumination 500lx, afterwards 10d illumination Time 5h, intensity of illumination 500-1000lx, incubation time 80d, the growth plant directly induced；3）Multiplication takes root while completes to cultivate：Will be through above-mentioned Fiber differentiation, acquired well-grown clump bud is according to plant size Plant division inoculation is carried out, high 0.3-0.5cm, the plantlet of blade 1-2 pieces are inoculated in bud proliferated culture medium, high about 0.5-1.0cm, The big plant of blade 2-3 pieces is inoculated in the culture medium for completing rooting process, light application time 8h/d, intensity of illumination 1500- 2000lx, Multiplying culture time 35-45d obtain multiplication and take root while complete to grow seedlings；4）The culture of test tube bottle seedling is completed：When culture of rootage test tube seedling grows supreme 5-6cm, root 3-5 items, leaf 3-5 pieces, the long 1.5- of leaf The germination of Herba Diploprora championii Seed inducement and quick breeding culture are completed during 2.5cm；6）It will complete culture of rootage test tube bottle seedling and be placed on natural light lower refining seedling 5-7 days, bottle cap hardening 1-2d is opened, to enhance test tube Seedling is to the adaptability of outdoor environment；Then taken out from blake bottle, clean the remaining medium for being attached to root system, move into humic Soil：Water plant mass ratio is 1:In the matrix of 2 mixing, moisturizing shading, temperature is controlled at 15-30 DEG C, and humidity is maintained at 75%- 85%, direct sunlight is avoided,；Plant adaptability gradually enhances after 2-3 weeks, obtains the complete seedling of nature robust growth.
- 2. a kind of Herba Diploprora championii sprouting and rooting method according to claim 1, it is characterised in that：The bud inducement cultivation base For：MS+1.0mg/L spends No. 1+1.0 gL of+0.5mg/LNAA+1.0g/L peptone+20g/L sucrose+5.8g/L agar powders of treasured-1 Activated carbon；PH value is 5.4-5.7.
- 3. a kind of Herba Diploprora championii sprouting and rooting method according to claim 1, it is characterised in that：The bud Multiplying culture Base：1/2MS+1.0g/L spends No. 2+0.5mg/LKT+0.1mg/LNAA+25g/L sucrose+5.6g/L agar powders+1.50gL of treasured-1 Activated carbon；PH value is 5.4-5.7.
- 4. a kind of Herba Diploprora championii sprouting and rooting method according to claim 1, it is characterised in that：The training of the rooting process Support base for 1/2MS+1.0g/L spend precious No. 2+0.5mg/LKT+0.1mg/LNAA+25g/L sucrose+5.6g/L agar powders+ 1.50g·L-1Activated carbon；PH value is 5.4-5.7.
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| CN104839027A (en) * | 2015-06-01 | 2015-08-19 | 福建农林大学 | Inducing method for maguey bulblet |
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| CN104839027A (en) * | 2015-06-01 | 2015-08-19 | 福建农林大学 | Inducing method for maguey bulblet |
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