CN106069782A - The rooting method for tissue culture of Tamarix jintaenia - Google Patents
The rooting method for tissue culture of Tamarix jintaenia Download PDFInfo
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- CN106069782A CN106069782A CN201610637864.3A CN201610637864A CN106069782A CN 106069782 A CN106069782 A CN 106069782A CN 201610637864 A CN201610637864 A CN 201610637864A CN 106069782 A CN106069782 A CN 106069782A
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- jintaenia
- tamarix
- outer implant
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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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- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
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- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
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Abstract
The invention discloses the rooting method for tissue culture of Tamarix jintaenia, including: (1) selects Tamarix jintaenia to give birth to the twig outer implant as root culture then；(2) outer implant is cut into the stage casing of a length of 10 15cm, does deep layer after rinsing out the silt on surface and clean；(3) on superclean bench, external implant carries out sterilization treatment；(4) outer implant is cut into the segment of long 1.5 2.5cm, is inoculated in root media (1/2MS+IBA 0.4mg/l+ sucrose 25g/l)；(5) condition of culture is set: temperature 24 ± 2 DEG C, intensity of illumination 1500 2000lx, light application time 10 14h/d.The invention have benefit that: Tamarix jintaenia not only root of hair is early, i.e. starting root of hair for the first time at the 5th day, and it is fast to take root, rootage duration foreshortens to 15 21 days, after 21 days, average root length reaches about 9cm, root is longer, and the par of root reaches 5, and Rhizoma Imperatae is many, rooting rate reaches 100%, top growing way is good, and seedling cost is substantially reduced, it is achieved that the industrialization of Tamarix jintaenia produces.
Description
Technical field
The present invention relates to the rooting method of a kind of plant, be specifically related to the rooting method for tissue culture of a kind of Tamarix jintaenia,
Belong to vegetative propagation technique field.
Background technology
Plant tissue culture is the nothing isolating the tissue, organ or cell, the protoplast etc. that suit the requirements from plant
Sexual reproduction method, i.e. by sterile working, carries out cultivating obtain complete regenerated plant or produce tool under manual control condition
There is the reproduction technique of other products of economic worth.
In the reproductive process of most of nursery stocks, traditional sexual propagation can also reach the most numerous purpose of taking root, but
The original merit of malleable pistillate parent in reproductive process, makes offspring morph.
As current widely used asexual reproduction method, tissue culture is not limited by geographical environment and seasonal conditions,
It is avoided that plant occurs trait segregation in growth course；Additionally, the method has, proliferative speed is fast, low cost, survival rate are high,
The features such as seedling speed is fast, solve the series of problems occurred in sexual propagation, thus obtain a large amount of sterile rootage Seedling.
Tamarix jintaenia (Tama Tamarix jintaenia) belongs to Salicaceae Salix, is distributed mainly on Hexi Corridor In Gansu gold
Dune slack near tower county.This seeds inflorescence is dense, the purplish red beauty of pattern, and branch and leaf are dark green, can make residential area, Desert Area attached
Close up reward and desert-control tree.Tamarix jintaenia happiness light is weak to the moon, at the many undergrowths in place that shelter from heat or light.Well developed root system, the most resistance to dry but also
Water-fast wet, wind loading rating is strong, salt tolerant alkaline earth, can on the salt-soda soil of salinity 1.2% normal growth, to the Desert Regions betterment of land
There is well effect.
But in recent years, owing to areal area landform, environment and weather conditions are the most changeable, Tamarix jintaenia natural regeneration is by pole
Big restriction, seedling reduces the most increasingly.At present, researcher mainly takes cottage method to breed this tree, and in this mistake
The problem such as occur in that in journey that cutting is difficult to preserve, cutting survival rate is low, seedling speed is slow after cuttage.
Therefore, it is badly in need of taking new vegetative method to solve this problem.
Summary of the invention
It is an object of the invention to provide the rooting method for tissue culture of a kind of Tamarix jintaenia, the method can make Tamarix jintaenia
Within a short period of time breeds fast and efficiently, thus creates higher economic benefit.
In order to realize above-mentioned target, the present invention adopts the following technical scheme that:
The rooting method for tissue culture of a kind of Tamarix jintaenia, it is characterised in that comprise the following steps:
Step1: select Tamarix jintaenia to give birth to the twig outer implant as root culture then；
Step2: outer implant is cut into the stage casing of a length of 10-15cm, does deep layer and cleans after rinsing out the silt on surface；
Step3: the outer implant cleaned up is placed on superclean bench, carries out sterilization treatment；
Step4: sterilized outer implant is cut into the segment of long 1.5-2.5cm, is inoculated in root media, aforementioned life
Root culture medium is: 1/2MS+IBA 0.4mg/l+ sucrose 25g/l, and pH value is 5.8-6.0；
Step5: postvaccinal outer implant cultivated, condition of culture is: temperature 24 ± 2 DEG C, intensity of illumination 1500-
2000lx, light application time 10-14h/d.
The rooting method for tissue culture of aforesaid Tamarix jintaenia, it is characterised in that in Step1, under fair weather
The outer implant of noon clip.
The rooting method for tissue culture of aforesaid Tamarix jintaenia, it is characterised in that in Step2, external implant is carried out deeply
The method that layer cleans is:
First cleaning branch with detergent, foams of washing powder dips a Tween 80 with Glass rod after rinsing well and carries out deep layer
Clean, afterwards outer implant is placed under flowing water flushing 2-6 hour.
The rooting method for tissue culture of aforesaid Tamarix jintaenia, it is characterised in that in Step3, external implant is gone out
The method that bacterium processes is:
First with 70% soak with ethanol 10-15s, then with aseptic water washing 3-4 time, then the mercuric chloride immersion with 0.1%
2min, finally rinses 5-7 time with sterilized water again.
The rooting method for tissue culture of aforesaid Tamarix jintaenia, it is characterised in that in Step4, outer implant is cut into segment
Before, first base portion otch is wiped out, then blot the moisture of outer planting surface.
The rooting method for tissue culture of aforesaid Tamarix jintaenia, it is characterised in that in Step4, outside every bottle graft kind one
Implant.
The invention have benefit that: when using the method for the present invention to cultivate Tamarix jintaenia, Tamarix jintaenia not only root of hair
Early, i.e. starting root of hair for the first time at the 5th day, and it is fast to take root, rootage duration foreshortens to 15-21 days, and after 21 days, average root is up to
To about 9cm, root is longer, and the par of root reaches 5, and Rhizoma Imperatae is many, and rooting rate reaches 100%, and top growing way is good, and nursery becomes
Originally it is substantially reduced, it is achieved that the industrialization of Tamarix jintaenia produces.
Accompanying drawing explanation
Fig. 1 is that outer implant cultivates the growing state figure of root after 28d in JS1 culture medium；
Fig. 2 is that outer implant cultivates the growing state figure of plant after 28d in JS1 culture medium；
Fig. 3 is that outer implant cultivates the growing state figure of root after 28d in JS2 culture medium；
Fig. 4 is that outer implant cultivates the growing state figure of plant after 28d in JS2 culture medium；
Fig. 5 is that outer implant cultivates the growing state figure of root after 28d in JS3 culture medium；
Fig. 6 is that outer implant cultivates the growing state figure of root after 28d in JS4 culture medium；
Fig. 7 is that outer implant cultivates the growing state figure of plant after 28d in JS4 culture medium；
Fig. 8 is that outer implant cultivates root and the growing state figure of plant after 28d in JS5 culture medium；
Fig. 9 is that outer implant cultivates root and the growing state figure of plant after 28d in JS6 culture medium；
Figure 10 is that outer implant cultivates root and plant strain growth situation map after 28d in JS7 culture medium；
Figure 11 is that outer implant cultivates the growing state figure of root after 28d in JS8 culture medium；
Figure 12 is that outer implant cultivates the growing state figure of plant after 28d in JS8 culture medium；
Figure 13 is that outer implant cultivates the growing state figure of root after 28d in JS9 culture medium；
Figure 14 is that outer implant cultivates root and plant strain growth situation map after 28d in JS10 culture medium；
Figure 15 is that outer implant cultivates root and plant strain growth situation map after 28d in JS11 culture medium；
Figure 16 is that outer implant cultivates the growing state figure of root after 28d in JS12 culture medium；
Figure 17 is that outer implant cultivates the growing state figure of plant after 28d in JS12 culture medium；
Figure 18 is that outer implant cultivates the growing state figure of root after 28d in JS13 culture medium；
Figure 19 is that outer implant cultivates the growing state figure of plant after 28d in JS13 culture medium.
Detailed description of the invention
Below in conjunction with the drawings and specific embodiments, the present invention made concrete introduction.
One, the selection of outer implant
Selecting suitable outer implant is the basic steps of tissue culture.
The present invention selects Tamarix jintaenia to give birth to the twig outer implant as root culture then, cuts not in the afternoon of fair weather
Outer implant with the age of stand.
Two, the cleaning of outer implant
1, preliminary flushing
In indoor, first the outer implant gathered is cut into the segment of a length of 10-15cm, rinses surface with tap water afterwards
Silt.
2, deep layer is cleaned
First cleaning branch with detergent, foams of washing powder dips a Tween 80 with Glass rod after rinsing well and carries out deep layer
Clean, afterwards outer implant is placed under flowing water flushing 2-6 hour.
Three, the sterilizing of outer implant and clip
1, sterilizing
The outer implant cleaned up is placed on superclean bench, first with 70% soak with ethanol 10-15s, then use nothing
Bacterium water rinses 3-4 time, then soaks 2min with the mercuric chloride of 0.1%, finally uses aseptic water washing 5-7 time.
2, clip
Sterilized outer implant base portion otch is wiped out, blots the moisture of outer planting surface with sterilized filter paper, be cut into
The segment of long 1.5-2.5cm, stand-by.
Four, root culture
1, screening culture medium
In the present invention, the root culture of Tamarix jintaenia with MS as minimal medium (wherein a great number of elements, trace element,
Iron salt, organic mother liquor content are all not changed in), and it is equipped with the indolebutyric acid (IBA) of variable concentrations, sucrose and activated carbon (AC).
Specific experiment process is divided into two stages, specific as follows:
First stage: screen the optimal IBA concentration of suitable Tamarix jintaenia root culture
JS1:MS；
JS2:1/2MS；
JS3:1/2MS+IBA 0.2mg/l+ sucrose 30g/l；
JS4:1/2MS+IBA 0.4mg/l+ sucrose 30g/l；
JS5:1/2MS+IBA 0.6mg/l+ sucrose 30g/l；
JS6:1/2MS+IBA 0.8mg/l+ sucrose 30g/l；
JS7:1/2MS+IBA 1.0mg/l+ sucrose 30g/l.
The pH value of the most each culture medium is all in the range of 5.8-6.0.
The outer implant segment of a length of 1.5-2.5cm is inoculated in above-mentioned culture medium, one outer implant of every bottle graft kind.
Condition of culture is arranged: temperature 24 ± 2 DEG C, intensity of illumination 1500-2000lx, light application time 10-14h/d.
Investigate the situation of taking root having inoculated outer implant after inoculation respectively, mainly include for the first time root of hair time, root length, root
Quantity, root system and plant strain growth situation.
Survey result is as follows:
By above table and accompanying drawing it was found that Tamarix jintaenia rootage duration the earliest and the best culture medium of plant growing way
For JS4:1/2MS+IBA 0.4mg/l+ sucrose 30g/l.
In this stage culture medium, the content of sucrose keeps 30g/l constant, simply adjusts the content of auxin IBA, finds
When adding the IBA of 0.4mg/l in 1/2MS culture medium, Tamarix jintaenia just can be taken root at 6d, and Rhizoma Imperatae is a lot, and top is planted
Strain is grown fine.
Second stage: adjust the culture medium prescription that sucrose is optimal with the concentration screening of activated carbon
Research shows, suitably reduces the content of sucrose and adds appropriate activated carbon (AC) beneficially tissue in the medium
The root culture of incubation China and foreign countries implant, therefore in second stage is cultivated, we are still with MS as minimal medium, first
On the basis of optimum growh element concentration (IBA0.4mg/l) that stage filters out, adjust the concentration of sucrose and activated carbon (AC), enter
One step screens the culture medium prescription of optimal hormone combinations.
JS8:1/2MS+IBA 0.4mg/l+ sucrose 5g/l；
JS9:1/2MS+IBA 0.4mg/l+ sucrose 10g/l；
JS10:1/2MS+IBA 0.4mg/l+ sucrose 15g/l；
JS11:1/2MS+IBA 0.4mg/l+ sucrose 20g/l；
JS12:1/2MS+IBA 0.4mg/l+ sucrose 25g/l；
JS13:1/2MS+IBA 0.4mg/l+ sucrose 30g/l；
JS14:1/2MS+IBA 0.4mg/l+ sucrose 30g/l+AC 0.5g/l；
JS15:1/2MS+IBA 0.4mg/l+ sucrose 30g/l+AC 1.0g/l；
JS16:1/2MS+IBA 0.4mg/l+ sucrose 30g/l+AC 2.0g/l；
JS17:1/2MS+IBA 0.4mg/l+ sucrose 30g/l+AC 5.0g/l.
The pH value of the most each culture medium is all in the range of 5.8-6.0.
The outer implant segment of a length of 1.5-2.5cm is inoculated in above-mentioned culture medium, one outer implant of every bottle graft kind.
Condition of culture is arranged: temperature 24 ± 2 DEG C, intensity of illumination 1500-2000lx, light application time 10-14h/d.
Investigate the situation of taking root having inoculated outer implant after inoculation respectively, mainly include for the first time root of hair time, root length, root
Quantity, root system and plant strain growth situation.
Survey result is as follows:
By above table and accompanying drawing it was found that Tamarix jintaenia is at JS12:1/2MS+IBA0.4mg/l+ sucrose 25g/l
When cultivating in culture medium, rooting efficiency is best, and relatively early (5d), main root is flourishing, and root hair is relatively mainly to show as rootage duration for the first time
Many, root top plant grows fine, and color is light green, and rooting rate is 100%.
By the cultivation of two different phases, our finishing screen have selected and is best suitable for the cultivation that the outer implant of Tamarix jintaenia is taken root
Based formulas, specific as follows:
1/2MS+IBA 0.4mg/l+ sucrose 25g/l (JS12).
2, inoculation
Being inoculated in the root media that finishing screen is elected by the outer implant segment of a length of 1.5-2.5cm, this is taken root
The formula of culture medium is: 1/2MS+IBA 0.4mg/l+ sucrose 25g/l, one outer implant of every bottle graft kind.
3, cultivate
Root culture condition setting: temperature 24 ± 2 DEG C, intensity of illumination 1500-2000lx, light application time 10-14h/d.
4, observe
Observed result: started root of hair (root of hair is early) for the first time at the 5th day, average root length reached about 9cm (life at the 21st day
Root is fast, root is longer), the par of root reaches 5, and Rhizoma Imperatae is many, and top growing way is good, and rooting rate reaches 100%.
As can be seen here, the asexual reproduction method that the present invention uses maintains the merit of original kind, is proposed
The culture medium being suitable for taking root make the outer implant of Tamarix jintaenia go out the root time early, neat, go out that root amount is big, Seedling of taking root is sturdy, leaf is tender
Green, growing way good.
Additionally, the inventive method also has, reproduction speed is fast, breeding coefficient big, convenient management, production cost are low, planting percent
High, cultivation cycle is short, can whole year production and be advantageously implemented factorial praluction, not by the feature limited season and advantage, can
Advantage is provided for large-scale industrialized production.
Due to this seeds well developed root system, there is again good Resistance to wind Erosion and Saline alkali tolerance, this invention is organized training and takes root
The research of culture medium can be desertification integrated control and Desert Regions betterment of land offer abundance, the source of seedling of high-quality.
In sum, method for tissue culture is the important method that asexual propagation field carries out species Fast-propagation, over entering year
Have been widely used and achieved with remarkable result in terms of nursery stock, flowers, medical material Fast-propagation, and utilize tissue culture's Rapid Rooting
Method in Tamarix jintaenia vegetative propagation also without reference to.The present invention utilizes the tissue culture side in asexual propagation field
Method, determines, by the screening in two stages, minimal medium formula and the hormone combinations that the training of applicable Tamarix jintaenia group is taken root, real
Show Tamarix jintaenia to breed fast and efficiently within a short period of time, breached and individually adopt during Tamarix jintaenia asexual propagation
By the method for cottage propagation, substantially increase reproduction speed and the effect of these seeds, efficiently solve Tamarix jintaenia take root difficulty
Problem, these species that are difficult to other kind of this platymiscium and cottage propagation thereof to take root play well with reference to and reference function.
Become it should be noted that above-described embodiment limits the present invention, all employing equivalents or equivalence the most in any form
The technical scheme that the mode changed is obtained, all falls within protection scope of the present invention.
Claims (6)
1. the rooting method for tissue culture of a Tamarix jintaenia, it is characterised in that comprise the following steps:
Step1: select Tamarix jintaenia to give birth to the twig outer implant as root culture then；
Step2: outer implant is cut into the stage casing of a length of 10-15cm, does deep layer and cleans after rinsing out the silt on surface；
Step3: the outer implant cleaned up is placed on superclean bench, carries out sterilization treatment；
Step4: sterilized outer implant is cut into the segment of long 1.5-2.5cm, is inoculated in root media, described in take root training
Foster base is: 1/2MS+IBA 0.4mg/l+ sucrose 25g/l, and pH value is 5.8-6.0；
Step5: postvaccinal outer implant cultivated, condition of culture is: temperature 24 ± 2 DEG C, intensity of illumination 1500-
2000lx, light application time 10-14h/d.
The rooting method for tissue culture of Tamarix jintaenia the most according to claim 1, it is characterised in that in Step1, fine
The outer implant of clip in afternoon of bright weather.
The rooting method for tissue culture of Tamarix jintaenia the most according to claim 1, it is characterised in that in Step2, externally
Implant carries out the method for deep layer cleaning:
First cleaning branch with detergent, foams of washing powder dips a Tween 80 with Glass rod after rinsing well, and to carry out deep layer clear
Wash, afterwards outer implant is placed under flowing water flushing 2-6 hour.
The rooting method for tissue culture of Tamarix jintaenia the most according to claim 1, it is characterised in that in Step3, externally
Implant carries out the method for sterilization treatment:
First with 70% soak with ethanol 10-15s, then with aseptic water washing 3-4 time, then the mercuric chloride immersion 2min with 0.1%,
Rinse again 5-7 time with sterilized water afterwards.
The rooting method for tissue culture of Tamarix jintaenia the most according to claim 1, it is characterised in that in Step4, outer planting
Before body is cut into segment, first base portion otch is wiped out, then blot the moisture of outer planting surface.
The rooting method for tissue culture of Tamarix jintaenia the most according to claim 1, it is characterised in that in Step4, every bottle
Inoculate an outer implant.
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Cited By (1)
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| CN108739235A (en) * | 2018-05-18 | 2018-11-06 | 句容市茂润苗木有限公司 | A kind of weeping willow seedling culture base and propagation method |
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| CN105052738A (en) * | 2015-07-31 | 2015-11-18 | 青岛正杰实业有限公司 | Tamarix chinensis tissue culture and rapid propagation method |
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| CN105052738A (en) * | 2015-07-31 | 2015-11-18 | 青岛正杰实业有限公司 | Tamarix chinensis tissue culture and rapid propagation method |
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| CN108739235A (en) * | 2018-05-18 | 2018-11-06 | 句容市茂润苗木有限公司 | A kind of weeping willow seedling culture base and propagation method |
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