CN106069782B - 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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- CN106069782B CN106069782B CN201610637864.3A CN201610637864A CN106069782B CN 106069782 B CN106069782 B CN 106069782B CN 201610637864 A CN201610637864 A CN 201610637864A CN 106069782 B CN106069782 B CN 106069782B
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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 the rooting method for tissue culture of Tamarix jintaenia, including：(1) Tamarix jintaenia is selected to give birth to explant of the spray as culture of rootage then；(2) explant is cut into the stage casing that length is 10 15cm, deep layer cleaning is done after rinsing out the silt on surface；(3) sterilization treatment is carried out to explant on superclean bench；(4) explant is cut into long 1.5 2.5cm segment, is inoculated in root media (1/2MS+IBA 0.4mg/l+ sucrose 25g/l)；(5) condition of culture is set：24 ± 2 DEG C of temperature, the 2000lx of intensity of illumination 1500, the 14h/d of light application time 10.The present invention is advantageous in that：Not only root of hair is early for Tamarix jintaenia, started first time root of hair at the 5th day, and it is fast to take root, and rootage duration foreshortens to 15 21 days, average root length reaches 9cm or so after 21 days, root is longer, and the par of root reaches 5, and hair root is more, rooting rate reaches 100%, top growing way is good, and seedling cost substantially reduces, and realizes the industrialization production of Tamarix jintaenia.
Description
Technical field
The present invention relates to a kind of rooting method of plant, and in particular to a kind of rooting method for tissue culture of Tamarix jintaenia,
Belong to vegetative propagation technique field.
Background technology
Plant Tissue Breeding is that the nothing of the tissue to suit the requirements, organ or cell, protoplast etc. is isolated from plant
Sexual reproduction method, i.e., by sterile working, cultivated under manual control condition to obtain complete regenerated plant or production tool
There is the reproduction technique of other products of economic value.
In the reproductive process of most of nursery stocks, traditional generative propagation can also reach fast numerous purpose of taking root, but
The original merit of malleable pistillate parent, makes offspring morph in reproductive process.
As current widely used asexual reproduction method, tissue cultures are not limited by geographical environment and seasonal conditions,
It is avoided that trait segregation occurs in growth course in plant；In addition, this method has, proliferative speed is fast, cost is low, survival rate is high,
The features such as seedling speed is fast, solves a series of problems occurred in generative propagation, so as to obtain a large amount of sterile rootage seedlings.
Tamarix jintaenia (Tama Tamarix jintaenia) belongs to Salicaceae Salix, is distributed mainly on Hexi Corridor In Gansu gold
Dune slack near tower county.The seeds inflorescence is dense, and the purplish red beauty of pattern, branches and leaves are dark green, and it is attached can to make Desert Area residential area
Close up is appreciated and desert-control tree.Tamarix jintaenia light not shade tolerant, in the more undergrowths in place that shelter from heat or light.Well developed root system, it is not only resistance to dry but also
Water-fast wet, wind loading rating is strong, salt tolerant alkaline earth, can on the salt-soda soil of salt content 1.2% normal growth, to the Desert Regions betterment of land
There is good effect.
But in recent years, because distributed area landform, environment and weather conditions are badly changeable, Tamarix jintaenia natural regeneration is by pole
Big limitation, seedling are just reduced increasingly.At present, researcher mainly takes cottage method to breed the tree, and mistake herein
Occurs the problems such as cuttings is difficult to preserve, cuttings survival rate is low, seedling speed is slow after cuttage in journey.
Therefore, it is badly in need of taking the method for new vegetative propagation to solve this problem.
The content of the invention
It is an object of the invention to provide a kind of rooting method for tissue culture of Tamarix jintaenia, this method can make Tamarix jintaenia
Fast and efficiently breed within a short period of time, so as to create higher economic benefit.
In order to realize above-mentioned target, the present invention adopts the following technical scheme that：
A kind of rooting method for tissue culture of Tamarix jintaenia, it is characterised in that comprise the following steps：
Step1：Tamarix jintaenia is selected to give birth to explant of the spray as culture of rootage then；
Step2：Explant is cut into the stage casing that length is 10-15cm, deep layer cleaning is done after rinsing out the silt on surface；
Step3：The explant cleaned up is placed on superclean bench, carries out sterilization treatment；
Step4：Sterilized explant is cut into long 1.5-2.5cm segment, is inoculated in root media, foregoing life
Root culture medium is：1/2MS+IBA 0.4mg/l+ sucrose 25g/l, pH value 5.8-6.0；
Step5：Explant after inoculation is cultivated, condition of culture is：24 ± 2 DEG C of temperature, intensity of illumination 1500-
2000lx, light application time 10-14h/d.
The rooting method for tissue culture of foregoing Tamarix jintaenia, it is characterised in that in Step1, under fair weather
Noon clip explant.
The rooting method for tissue culture of foregoing Tamarix jintaenia, it is characterised in that in Step2, carried out to explant deep
Layer cleaning method be：
Branch first is cleaned with washing powder, foams of washing powder dips a drop Tween 80 with glass bar after rinsing well and carries out deep layer
Cleaning, explant is placed in flushing 2-6 hours under flowing water afterwards.
The rooting method for tissue culture of foregoing Tamarix jintaenia, it is characterised in that in Step3, gone out to explant
Bacterium processing method be：
10-15s first is soaked with 70% ethanol, then with aseptic water washing 3-4 times, then is soaked with 0.1% mercuric chloride
2min, finally rinsed again 5-7 times with sterilized water.
The rooting method for tissue culture of foregoing Tamarix jintaenia, it is characterised in that in Step4, explant is cut into segment
Before, first base portion otch is wiped out, then blots the moisture on explant surface.
The rooting method for tissue culture of foregoing Tamarix jintaenia, it is characterised in that in Step4, every bottle of inoculation one is outer
Implant.
The present invention is advantageous in that：Using the present invention method culture Tamarix jintaenia when, Tamarix jintaenia not only root of hair
It is early, started first time root of hair at the 5th day, and also it is fast to take root, and average root is up to after rootage duration foreshortens to 15-21 days, 21 days
Arrive 9cm or so, root is longer, and the par of root reaches 5, and hair root is more, and rooting rate reaches 100%, and top growing way is good, nursery into
Originally substantially reduce, realize the industrialization production of Tamarix jintaenia.
Brief description of the drawings
Fig. 1 is the growing state figure that explant cultivates root after 28d in JS1 culture mediums；
Fig. 2 is the growing state figure that explant cultivates plant after 28d in JS1 culture mediums；
Fig. 3 is the growing state figure that explant cultivates root after 28d in JS2 culture mediums；
Fig. 4 is the growing state figure that explant cultivates plant after 28d in JS2 culture mediums；
Fig. 5 is the growing state figure that explant cultivates root after 28d in JS3 culture mediums；
Fig. 6 is the growing state figure that explant cultivates root after 28d in JS4 culture mediums；
Fig. 7 is the growing state figure that explant cultivates plant after 28d in JS4 culture mediums；
Fig. 8 is that explant cultivates root and the growing state figure of plant after 28d in JS5 culture mediums；
Fig. 9 is that explant cultivates root and the growing state figure of plant after 28d in JS6 culture mediums；
Figure 10 is that explant cultivates root and plant strain growth situation map after 28d in JS7 culture mediums；
Figure 11 is the growing state figure that explant cultivates root after 28d in JS8 culture mediums；
Figure 12 is the growing state figure that explant cultivates plant after 28d in JS8 culture mediums；
Figure 13 is the growing state figure that explant cultivates root after 28d in JS9 culture mediums；
Figure 14 is that explant cultivates root and plant strain growth situation map after 28d in JS10 culture mediums；
Figure 15 is that explant cultivates root and plant strain growth situation map after 28d in JS11 culture mediums；
Figure 16 is the growing state figure that explant cultivates root after 28d in JS12 culture mediums；
Figure 17 is the growing state figure that explant cultivates plant after 28d in JS12 culture mediums；
Figure 18 is the growing state figure that explant cultivates root after 28d in JS13 culture mediums；
Figure 19 is the growing state figure that explant cultivates plant after 28d in JS13 culture mediums.
Embodiment
Make specific introduce to the present invention below in conjunction with the drawings and specific embodiments.
First, the selection of explant
Select the basic steps that suitable explant is tissue cultures.
Present invention selection Tamarix jintaenia gives birth to explant of the spray as culture of rootage then, is cut not in the afternoon of fair weather
With the explant at the age of stand.
2nd, the cleaning of explant
1st, preliminary flushing
Indoors, the explant of collection is first cut into the segment that length is 10-15cm, rinses surface with running water afterwards
Silt.
2nd, deep layer is cleaned
Branch first is cleaned with washing powder, foams of washing powder dips a drop Tween 80 with glass bar after rinsing well and carries out deep layer
Cleaning, explant is placed in flushing 2-6 hours under flowing water afterwards.
3rd, the sterilizing of explant and clip
1st, sterilize
The explant cleaned up is placed on superclean bench, first 10-15s is soaked with 70% ethanol, then with nothing
Bacterium water rinses 3-4 times, then soaks 2min with 0.1% mercuric chloride, finally with aseptic water washing 5-7 times.
2nd, clip
Sterilized explant base portion otch is wiped out, the moisture on explant surface is blotted with sterilized filter paper, is cut into
Long 1.5-2.5cm segment, it is stand-by.
4th, culture of rootage
1st, screening and culturing medium
In the present invention, the culture of rootage of Tamarix jintaenia using MS as minimal medium (wherein a great number of elements, trace element,
Molysite, organic mother liquor content do not change), and it is equipped with the indolebutyric acid (IBA), sucrose and activated carbon (AC) of various concentrations.
Specific experiment process is divided into two stages, specific as follows：
First stage：Screen the optimal IBA concentration of suitable Tamarix jintaenia culture of rootage
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 each culture medium is in the range of 5.8-6.0 above.
The explant segment that length is 1.5-2.5cm is inoculated in above-mentioned culture medium, one explant of every bottle of inoculation.
Condition of culture is set：24 ± 2 DEG C of temperature, intensity of illumination 1500-2000lx, light application time 10-14h/d.
The situation of taking root for being inoculated with explant is investigated after inoculation respectively, it is main including root of hair time first time, root long, root
Quantity, root system and plant strain growth situation.
Investigation result is as follows：
By above table and accompanying drawing it was found that the culture medium that Tamarix jintaenia rootage duration is earliest and plant growing way is best
For JS4：1/2MS+IBA 0.4mg/l+ sucrose 30g/l.
The content of sucrose keeps 30g/l constant in this stage culture medium, simply adjusts auxin IBA content, finds
When adding 0.4mg/l IBA in 1/2MS culture mediums, Tamarix jintaenia is taken root in 6d cans, and hair root is a lot, and top is planted
Strain is grown fine.
Second stage：The optimal culture medium prescription of the concentration screening of adjustment sucrose and activated carbon
Research shows, the appropriate content for reducing sucrose and adds appropriate activated carbon (AC) in the medium and is advantageous to tissue
The culture of rootage of explant in incubation, therefore we are still using MS as minimal medium in second stage culture, first
On the basis of the optimum growh element concentration (IBA0.4mg/l) that stage filters out, the concentration of adjustment 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 each culture medium is in the range of 5.8-6.0 above.
The explant segment that length is 1.5-2.5cm is inoculated in above-mentioned culture medium, one explant of every bottle of inoculation.
Condition of culture is set：24 ± 2 DEG C of temperature, intensity of illumination 1500-2000lx, light application time 10-14h/d.
The situation of taking root for being inoculated with explant is investigated after inoculation respectively, it is main including root of hair time first time, root long, root
Quantity, root system and plant strain growth situation.
Investigation result is as follows：
By above table and accompanying drawing it was found that Tamarix jintaenia is in JS12：1/2MS+IBA0.4mg/l+ sucrose 25g/l
When being cultivated in culture medium, rooting efficiency is best, is mainly shown as that first time rootage duration is relatively early (5d), and main root is flourishing, root hair compared with
More, root top plant grows fine, and color is light green, rooting rate 100%.
By the culture of two different phases, our finishing screens have selected the culture that most suitable Tamarix jintaenia explant is taken root
Based formulas, it is specific as follows：
1/2MS+IBA 0.4mg/l+ sucrose 25g/l (JS12).
2nd, it is inoculated with
The explant segment that length is 1.5-2.5cm is inoculated in the root media finally screened, this is taken root
The formula of culture medium is：1/2MS+IBA 0.4mg/l+ sucrose 25g/l, one explant of every bottle of inoculation.
3rd, cultivate
Culture of rootage condition setting：24 ± 2 DEG C of temperature, intensity of illumination 1500-2000lx, light application time 10-14h/d.
4th, observe
Observe result：Started first time root of hair (root of hair is early) at the 5th day, it is (raw to reach 9cm or so for average root long at the 21st day
Root is fast, root is longer), the par of root reaches 5, and hair root is more, 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 most
The culture medium for being adapted to take root cause Tamarix jintaenia explant go out the root time it is early, it is neat, go out that root amount is big, rooted seedling is sturdy, leaf is tender
It is green, growing way is good.
In addition, the inventive method also has, reproduction speed is fast, breeding coefficient is big, management is convenient, production cost is low, planting percent
It is high, cultivation cycle is short, can whole year production and the characteristics of be advantageously implemented factorial praluction, do not limited by season and advantage, can
Advantage is provided for large-scale industrialized production.
Due to the seeds well developed root system, there is good Resistance to wind Erosion and Saline alkali tolerance again, tissue culture is taken root in the invention
The research of culture medium can provide sufficient, high-quality source of seedling for desertification integrated control and the Desert Regions betterment of land.
In summary, method for tissue culture is that vegetative propagation field carries out the important method that species are quickly bred, over entering year
Had been widely used in nursery stock, flowers, the quick proliferation of medicinal material and obtained remarkable result, and utilize tissue cultures Rapid Rooting
Method in Tamarix jintaenia vegetative propagation also without reference to.The present invention utilizes the tissue cultures side in vegetative propagation field
Method, the minimal medium formula and hormone combinations that suitable Tamarix jintaenia tissue culture takes root are determined by the screening in two stages, it is real
Tamarix jintaenia is showed fast and efficiently to breed within a short period of time, has breached and individually adopted during Tamarix jintaenia vegetative propagation
With the method for cutting propagation, the reproduction speed and effect of the seeds are substantially increased, Tamarix jintaenia is efficiently solved and takes root hardly possible
Problem, this species that are difficult to take root to the other kinds of the platymiscium and its cutting propagation play reference and reference function well.
It should be noted that the invention is not limited in any way for above-described embodiment, it is all to use equivalent substitution or equivalent change
The technical scheme that the mode changed is obtained, all falls within protection scope of the present invention.
Claims (6)
1. a kind of rooting method for tissue culture of Tamarix jintaenia, it is characterised in that comprise the following steps：
Step1：Tamarix jintaenia is selected to give birth to explant of the spray as culture of rootage then；
Step2：Explant is cut into the stage casing that length is 10-15cm, deep layer cleaning is done after rinsing out the silt on surface；
Step3：The explant cleaned up is placed on superclean bench, carries out sterilization treatment；
Step4：Sterilized explant is cut into long 1.5-2.5cm segment, is inoculated in root media, the training of taking root
Foster base is：1/2MS+IBA 0.4mg/l+ sucrose 25g/l, pH value 5.8-6.0；
Step5：Explant after inoculation is cultivated, condition of culture is：24 ± 2 DEG C of temperature, intensity of illumination 1500-
2000lx, light application time 10-14h/d.
2. the rooting method for tissue culture of Tamarix jintaenia according to claim 1, it is characterised in that in Step1, fine
The clip explant in afternoon of bright weather.
3. the rooting method for tissue culture of Tamarix jintaenia according to claim 1, it is characterised in that in Step2, externally
Implant carry out deep layer cleaning method be：
First branch is cleaned with washing powder, foams of washing powder with glass bar dips a drop Tween 80 after rinsing well, and to carry out deep layer clear
Wash, explant is placed in flushing 2-6 hours under flowing water afterwards.
4. the rooting method for tissue culture of Tamarix jintaenia according to claim 1, it is characterised in that in Step3, externally
Implant carry out sterilization treatment method be：
10-15s first is soaked with 70% ethanol, then with aseptic water washing 3-4 times, then with 0.1% mercuric chloride soaks 2min, most
Rinsed again 5-7 times with sterilized water afterwards.
5. the rooting method for tissue culture of Tamarix jintaenia according to claim 1, it is characterised in that in Step4, explant
Before body is cut into segment, first base portion otch is wiped out, then blots the moisture on explant surface.
6. the rooting method for tissue culture of Tamarix jintaenia according to claim 1, it is characterised in that in Step4, every bottle
It is inoculated with an explant.
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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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| "短枝型怪柳离体快繁技术研究;杨立文等;《中国观赏园艺研究进展》;20131231;第299页右栏第2段及表3，第301页右栏第2段 * |
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