CN106508678A - Tamarix taklamakanensis tissue culture and rooting method - Google Patents
Tamarix taklamakanensis tissue culture and rooting method Download PDFInfo
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- CN106508678A CN106508678A CN201610958954.2A CN201610958954A CN106508678A CN 106508678 A CN106508678 A CN 106508678A CN 201610958954 A CN201610958954 A CN 201610958954A CN 106508678 A CN106508678 A CN 106508678A
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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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- 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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Abstract
The invention provides a Tamarix taklamakanensis tissue culture and rooting method. The method comprises the following steps: selecting an explant; pretreating the explant; sterilizing and scissoring the explant, and performing rooting culture, wherein the formula of a culture medium for rooting culture is 1/2 MS basal culture medium+0.2g/L IBA+15-30g/L sucrose, and the pH value is 5.8-6.0; and optimizing the culture conditions. According to the method provided by the invention, good characters of the original variety can be kept; the Tamarix taklamakanensis explant can root early with the effects of regular rooting time and large root amount; and after rooting, seedlings are robust, leaves are light green, and the growing state is good. Besides, the method provided by the invention has the characteristics and advantages of high propagation speed, large propagation coefficient, convenient management, low production cost, high planting percent and short culture period, can realize annual production, is beneficial to implementing industrialized production and is not restricted by seasons, thereby providing favorable conditions for large-scale industrialized production.
Description
Technical field
The invention belongs to technical field of tissue culture, and in particular to the raw Ramulus et Folium Tamariciss rooting method for tissue culture of a planting sand.
Background technology
Husky raw Ramulus et Folium Tamariciss（Tama Tamarix jintaenia）Category Salicaceae Salix, is China's Desert Regions endemic species, day
So distribution is little, and the Kumtag for being only distributed in the dry desert of Xinjiang of China Tarim Basin carat agate and Basin Eastern is husky
Desert, extends to the west edge of Dunhuang, Gansu Province, is born in away from inside the drift sand around having water riverbed and lake basin, while husky raw Ramulus et Folium Tamariciss
Ecological environment of distribution is harsh, is faced with threat in imminent danger, is listed in country and treasures endangered plants register.Husky raw Ramulus et Folium Tamariciss are in China arid desert
The annidation in area is extremely strong, and ecologic niche and its ecological significance are great.But, the raw Ramulus et Folium Tamariciss of sand of current China's natural distributed by
Husky table soil salt is gradually reduced and between moving dunes in level of ground water constantly to increase, natural regeneration is extremely restricted, and grows directly from seeds
Seedling is just reduced increasingly, implements husky raw Ramulus et Folium Tamariciss introducing and planting, is that Desert Regions are defended and controlled sand the recovery weight of plant selection-breeding and Ramulus et Folium Tamariciss shrubbery
Offer new material is built, the phylogeny to studying Central Asia ermophyte Flora and this category has scientific meaning.
Plant tissue culture is the nothing for isolating the tissue, organ or cell, the protoplast that suit the requirements etc. from plant
Sexual reproduction method, i.e., by sterile working, cultivated to obtain complete regenerated plant or production 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 its fast numerous purpose of taking root, but
The original merit of malleable pistillate parent in reproductive process so as to which offspring morphs.
Used 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, the series of problems occurred in solving sexual propagation, so that obtain a large amount of sterile rootage Seedlings.Cause
This, the method can make husky raw Ramulus et Folium Tamariciss reach the purpose of quick, efficient breeding within a short period of time, create higher economic effect
Benefit.
Mainly based on asexual propagation, it is premise and the basis of cutting propagation that cutting is survived, preserved to husky raw Ramulus et Folium Tamariciss, understands the palm
Husky raw Ramulus et Folium Tamariciss asexual propagation characteristic is held, asexual propagation shoot survival percent and storage rate is improved, the life to the raw Ramulus et Folium Tamariciss apogamic seedling of sand
It is important that root study mechanism seems that pole has.Method for tissue culture is the important method that asexual propagation field carries out that species are quickly bred,
Have been widely used and achieved with remarkable result in nursery stock, flowers, the quick proliferation of medical material entering year, and utilize tissue culture rapid
The method of fast-growing root wood in the raw Ramulus et Folium Tamariciss vegetative propagation of sand is related to.
The content of the invention
In order to solve problems of the prior art, the invention provides the side of taking root of the raw Ramulus et Folium Tamariciss tissue culture of a planting sand
Method, promotes its method taken root to reach the purpose that husky raw Ramulus et Folium Tamariciss quickly breed using tissue culture.
The present invention provides a planting sand raw Ramulus et Folium Tamariciss rooting method for tissue culture, comprises the following steps：
（1）The selection of explant：Husky raw Ramulus et Folium Tamariciss are selected to give birth to twig then as the explant of root culture；
（2）Explant pre-treatment；
（3）The sterilizing of explant and clip；
（4）Root culture：Root culture culture medium prescription is：1/2MS basal medium+IBA 0.2g/L+ sucrose 15-30
g/L；PH value is 5.8-6.0；Root culture condition of culture is：24 ± 2 DEG C of temperature, intensity of illumination 1500-2000lx, light application time
10-14 h/d；Sucrose is not contained in the MS basal mediums, and agar concentration is 5g/L.
Preferably, step（2）Described in explant pre-treatment be to select the husky raw Ramulus et Folium Tamariciss of clip in afternoon of fair weather ought
Raw branch was first cut the explant of collection to 10-15cm indoors, was rushed with tap water afterwards as the explant of root culture year
Wash the silt on surface；Then branch is cleaned with detergent, dip a drop tween with Glass rod after foams of washing powder is rinsed well
80 carry out deep layer cleaning, and explant is placed in flushing 2-6 hours under flowing water afterwards.
Preferably, step（3）Described in the sterilizing of explant be that the explant that will be rinsed well is placed in superclean bench
On, first with 75% ethanol immersion 10-15s, with aseptic water washing 3-4 time, then with 0.1% mercuric chloride immersion 3-6 min, according to old
Tender degree is different, and old branch bar soak time is longer, is 5-6 min；Shoot soak time is short, is 3-4min, afterwards with aseptic
Water is rinsed 5-7 time.
Preferably, step（3）Described in the clip of explant be that sterilized explant base portion otch is wiped out, with
The filter paper of sterilizing blots the moisture on explant surface, is cut into long 1-2cm length the segment with bud, is inoculated in root media,
One explant of per bottle of inoculation.
Preferably, step（4）Described in root culture culture medium prescription be：1/2MS basal medium+IBA 0.2g/
L+ sucrose 15-20 g/L.
Preferably, step（4）Described in root culture culture medium prescription be：1/2MS basal medium+IBA 0.2g/
L+ sucrose 15g/L.
The present invention also provides a planting sand raw Ramulus et Folium Tamariciss culture medium for tissue culture, and the formula of the culture medium is：The training of 1/2MS bases
Foster base+IBA 0.2g/L+ sucrose 15-30g/L.
Preferably, the formula of the culture medium is：1/2MS basal medium+IBA 0.2g/L+ sucrose 15-20g/
L。
Preferably, the formula of the culture medium is：1/2MS basal medium+IBA 0.2g/L+ sucrose 15g/L.
Method for tissue culture of the present invention by using asexual propagation field, solves the problems, such as that the raw Ramulus et Folium Tamariciss of sand take root hardly possible, and
Proposition is best suitable for the culture medium prescription that explant is taken root.Concrete steps mainly include the selection of explant, the front place of explant inoculation
Take root after reason, the sterilizing of explant and clip, the configuration of root media and dispensing information, inoculation and root growth situation observation.
Culture of the present invention by three different phases, screens and discloses the concrete training for being adapted to that husky raw Ramulus et Folium Tamariciss tissue culture is taken root
Minimal medium and species that the culture medium prescription of foster base and its different hormone combinations, i.e. different phase are used, culture medium are added
Plant growth regulator, the concentration of sucrose, culture medium acid-base value（PH）, the external environment condition of explant culture, such as illumination,
Humiture etc..
Method of the present invention using tissue culture, determines suitable husky raw Ramulus et Folium Tamariciss tissue culture by the screening of three phases and takes root
Minimal medium formula and hormone combinations, breach the side for individually adopting cutting propagation during the raw Ramulus et Folium Tamariciss asexual propagation of sand
Method, substantially increases the reproduction speed and effect of the seeds, is difficult to thing of taking root to other kinds of the platymiscium and its cutting propagation
Plant and play reference and reference function well.
The asexual reproduction method that the present invention is adopted can keep the merit of original kind, and what is proposed is best suitable for what is taken root
Culture medium, husky raw Ramulus et Folium Tamariciss explant can be made compared with early taking root, and go out that the root time is neat, go out root amount greatly, take root after seedling is sturdy, leaf
Light green, growing way is good；In addition, the present invention carries out the asexual propagation of seeds difficult to take root using method for tissue culture, with reproduction speed
Hurry up, breeding coefficient is big, convenient management, low production cost, planting percent high, cultivation cycle is short, whole year production and can be advantageously implemented
Factorial praluction, the characteristics of do not limited by season and advantage, advantage can be provided for large-scale industrialized production.
Further, since husky raw Ramulus et Folium Tamariciss well developed root system, and with good weather-proof, the husky ability of burying and Saline alkali tolerance, therefore,
The invention is applied to the seedling that production practices can provide sufficient, high-quality for desertification integrated control and the Desert Regions betterment of land to come
Source.
Description of the drawings
Accompanying drawing is used for providing a further understanding of the present invention, and constitutes a part for description, the reality with the present invention
Applying example is used for explaining the present invention together, is not construed as limiting the invention.In the accompanying drawings：
Fig. 1-Fig. 8 is respectively using the experimental result of the husky raw Ramulus et Folium Tamariciss of different tissue culture medium (TCM) cultures.
Specific embodiment
Below example facilitates a better understanding of the present invention, but does not limit the present invention.Experiment in following embodiments
Method, if no special instructions, is conventional method.In following embodiments, test material used, if no special instructions, is city
Sell.
First, the experimentation of the raw Ramulus et Folium Tamariciss method for tissue culture rooting method of a planting sand of the invention is as follows：
1st, the selection of explant：
The selection of suitable explant is the basis of tissue culture, and the husky raw Ramulus et Folium Tamariciss current-year branch of present invention selection is used as root culture
Explant, study its tissue culture and take root situation.
2nd, explant pre-treatment：
The husky raw Ramulus et Folium Tamariciss current-year branch of clip in afternoon of fair weather is selected as the explant of root culture；
It is indoor first to cut the explant of collection to 10-15cm, rinse the silt on surface afterwards with tap water；
First branch is cleaned with detergent, dipping a drop Tween 80 after foams of washing powder is rinsed well with Glass rod carries out depth
Explant is placed in flushing 2-6 hours under flowing water by layer cleaning afterwards.
3rd, the sterilizing of explant and clip：
The sterilizing of explant：The explant rinsed well is placed on superclean bench, first 15s is soaked with 75% ethanol,
With aseptic water washing 3-4 time, then with 0.1% mercuric chloride immersion 3-6 min（It is different according to old tender degree, old branch bar soak time compared with
It is long, generally 5-6 min；Shoot soak time is short, generally 3-4min）, afterwards with aseptic water washing 5-7 time；
The clip of explant after sterilizing：Sterilized explant base portion otch is wiped out, is blotted with sterilized filter paper outer
The moisture on implant surface, is cut into long 1-2cm length the segment with bud, is inoculated in root media, one explant of per bottle of inoculation
Body.
4th, root culture：
The root culture of husky raw Ramulus et Folium Tamariciss culture medium based on MS in the present invention（Wherein a great number of elements, trace element, iron salt, have
Machine mother liquor content is not changed in, and without sucrose, agar concentration is 5g/L）, and it is equipped with the indolebutyric acid of variable concentrations（IBA）、
Sucrose, activated carbon（AC）, specific experiment process is divided into three phases, and each stage culture medium and its formula are as follows：
First stage：Minimal medium and optimum growh hormone concentration of the screening suitable for husky raw Ramulus et Folium Tamariciss root culture：
1：MS basal mediums；
2：0.1 g/L+ sucrose of MS basal mediums+IBA, 30 g/L；
3：0.2 g/L+ sucrose of MS basal mediums+IBA, 30 g/L；
4：0.3 g/L+ sucrose of MS basal mediums+IBA, 30 g/L；
5：0.4 g/L+ sucrose of MS basal mediums+IBA, 30 g/L；
6：0.5 g/L+ sucrose of MS basal mediums+IBA, 30 g/L；
7：1/2MS basal mediums；
8：0.1 g/L+ sucrose of 1/2MS basal mediums+IBA, 30 g/L；
9：0.2 g/L+ sucrose of 1/2MS basal mediums+IBA, 30 g/L；
10：0.3 g/L+ sucrose of 1/2MS basal mediums+IBA, 30 g/L；
11：0.4 g/L+ sucrose of 1/2MS basal mediums+IBA, 30 g/L；
12：30 g/L of 1/2MS basal medium+IBA0.5 g/L+ sucrose；
Second stage：The concentration that sucrose is adjusted on the basis of the optimal hormone concentration for filtering out in the first stage is further screened most
The culture medium prescription of good hormone combinations：
S1：0.2 g/L+ sucrose of 1/2MS basal mediums+IBA, 0 g/L；
S2：0.2 g/L+ sucrose of 1/2MS basal mediums+IBA, 5 g/L；
S3：0.2 g/L+ sucrose of 1/2MS basal mediums+IBA, 10 g/L；
S4：0.2 g/L+ sucrose of 1/2MS basal mediums+IBA, 15 g/L；
S5：0.2 g/L+ sucrose of 1/2MS basal mediums+IBA, 20 g/L；
S6：0.2 g/L+ sucrose of 1/2MS basal mediums+IBA, 25 g/L；
S7：0.2 g/L+ sucrose of 1/2MS basal mediums+IBA, 30 g/L；
S8：0.2 g/L+ sucrose of 1/2MS basal mediums+IBA, 35 g/L.
Phase III：The activated carbon of variable concentrations is added in the optimal medium that second stage is filtered out（AC）.
5th, take root situation after being inoculated with：
Investigate the situation of taking root for being inoculated with explant after inoculation respectively, mainly include root of hair time first time, root length, the number of root
Amount, root system and plant strain growth situation.
6th, condition of culture is arranged：24 ± 2 DEG C of temperature, intensity of illumination 1500-2000lx, light application time 10-14 h/d.
Each of the above cultivation stage using minimal medium be MS basal mediums, it is additional variety classes, different dense
The plant growth regulator of degree, sucrose, activated carbon, medium pH are 5.8-6.0.
2nd, the experimental result of the raw Ramulus et Folium Tamariciss method for tissue culture rooting method of a planting sand of the invention is：
The present invention selects husky raw Ramulus et Folium Tamariciss branch to take root explant as tissue culture first, is cut into and is about 1.5-2cm's after being sterilized
Segment, carries out the screening of tissue culture's root media in being inoculated in the root media of first stage, culture medium is with MS as base
Basal culture medium, and IBA, the sucrose of additional variable concentrations, as a result as shown in table 1.
1 first stage of table cultivates
As can be seen from Table 1, the first stage filters out husky raw Ramulus et Folium Tamariciss rootage duration earliest and the best culture medium of plant growing way is 9
Number culture medium：30 g of 1/2MS basal medium+IBA 0.2+ sucrose, may thereby determine that suitable husky raw Ramulus et Folium Tamariciss explant is external
The minimal medium of implant root culture is 1/2MS basal mediums, and hormone concentration is IBA 0.2mg/l, and based on this
Carry out the culture of second stage.The experimental result of second stage is as shown in table 2.
2 second stage culture of table
Research shows, sucrose in addition to the energy is played a part of, also with the effect for adjusting the differentiation of cultured tissue woody part, and
With the change of the sucrose concentration combined with growth hormone, in calluss, the relative amount of xylem and phloem there occurs very
Big change；Cell elongation and root cell growth, promotion that growth hormone IBA has to be stimulated cambial cell division, stimulate branch
Xylem, phloem cell differentiation, promote cutting root of hair, the effect of the morphogenesis of regulation calluss, are mainly used in mostly
The cuttage of number xylophyta rises rooting propagation, and the IBA of low concentration is preferable to explant rooting effect in tissue culture.Therefore
Still with MS as minimal medium in second stage culture, good No. 9 culture medium of being taken root with first stage incubation is
The minimal medium of two-stage root culture, i.e., based on IBA 0.2mg/l, adjust the content of sucrose in culture medium.As a result
It was found that when sucrose concentration is 15mg/l, husky raw Ramulus et Folium Tamariciss rooting efficiency preferably, is mainly shown as that first time rootage duration is more early, main
Root is flourishing, and root hair is more, and root top plant grows fine, and color is light green, and its rooting rate is 88.3%.
In the phase III, after adding variable concentrations activated carbon in the medium, culture medium is presented black, and rooting efficiency is not
Substantially, thus final choice does not add activated carbon in the medium.I.e. S4 culture medium prescriptions are optimum culture medium prescription.
After the raw Ramulus et Folium Tamariciss root media of above-mentioned sand and cultural method, the rootage duration of husky raw Ramulus et Folium Tamariciss foreshortens to 15-21
My god, after January, seedling growth is accelerated.
Embodiment 1
The raw Ramulus et Folium Tamariciss rooting method for tissue culture of sand of the present invention is comprised the following steps that：
1st, the selection of explant：
The selection of suitable explant is the basis of tissue culture, of the invention to select husky raw Ramulus et Folium Tamariciss to give birth to twig then as root culture
Explant, study its tissue culture and take root situation.
2nd, explant pre-treatment：
The husky raw Ramulus et Folium Tamariciss current-year branch of clip in afternoon of fair weather is selected as the explant of root culture；
The silt on surface first with the explant of collection is cut to 12cm, is rinsed afterwards in interior with tap water；
First branch is cleaned with detergent, dipping a drop Tween 80 after foams of washing powder is rinsed well with Glass rod carries out depth
Layer cleaning, is placed in explant under flowing water afterwards and rinses 4 hours.
3rd, the sterilizing of explant and clip：
The sterilizing of explant：The explant rinsed well is placed on superclean bench, first 12s is soaked with 75% ethanol,
With aseptic water washing 3 times, then with 0.1% mercuric chloride immersion 4min, afterwards with aseptic water washing 6 times；
The clip of explant after sterilizing：Sterilized explant base portion otch is wiped out, is blotted with sterilized filter paper outer
The moisture on implant surface, is cut into long 1.cm length the segment with bud, is inoculated in root media, one explant of per bottle of inoculation
Body.
4th, root culture：
Prescription of rooting medium is：0.2 g/L+ sucrose of 1/2MS basal mediums+IBA, 15 g/L, pH value is 5.9；Culture
Condition is：24 DEG C of temperature, intensity of illumination 1800lx, light application time 12h/d；Sucrose, agar is not contained wherein in MS basal mediums
Concentration is 5g/L.
Embodiment 2
The raw Ramulus et Folium Tamariciss rooting method for tissue culture of sand of the present invention is comprised the following steps that：
1st, the selection of explant：
The selection of suitable explant is the basis of tissue culture, of the invention to select husky raw Ramulus et Folium Tamariciss to give birth to twig then as root culture
Explant, study its tissue culture and take root situation.
2nd, explant pre-treatment：
The husky raw Ramulus et Folium Tamariciss of clip in afternoon of fair weather are selected to give birth to twig then as the explant of root culture；
The silt on surface first with the explant of collection is cut to 10cm, is rinsed afterwards in interior with tap water；
First branch is cleaned with detergent, dipping a drop Tween 80 after foams of washing powder is rinsed well with Glass rod carries out depth
Layer cleaning, is placed in explant under flowing water afterwards and rinses 6 hours.
3rd, the sterilizing of explant and clip：
The sterilizing of explant：The explant rinsed well is placed on superclean bench, first 10s is soaked with 75% ethanol,
With aseptic water washing 3 times, then 6 min are soaked with 0.1% mercuric chloride, afterwards with aseptic water washing 7 times；
The clip of explant after sterilizing：Sterilized explant base portion otch is wiped out, is blotted with sterilized filter paper outer
The moisture on implant surface, is cut into long 2cm length the segment with bud, is inoculated in root media, one explant of per bottle of inoculation.
4th, root culture：
Prescription of rooting medium is：0.2 g/L+ sucrose of 1/2MS basal mediums+IBA, 20 g/L, pH value is 5.8；Culture
Condition is：22 DEG C of temperature, intensity of illumination 1500lx, light application time 14h/d；Sucrose, agar is not contained wherein in MS basal mediums
Concentration is 5g/L.
Embodiment 3
The raw Ramulus et Folium Tamariciss rooting method for tissue culture of sand of the present invention is comprised the following steps that：
1st, the selection of explant：
The selection of suitable explant is the basis of tissue culture, of the invention to select husky raw Ramulus et Folium Tamariciss to give birth to twig then as root culture
Explant, study its tissue culture and take root situation.
2nd, explant pre-treatment：
The husky raw Ramulus et Folium Tamariciss of clip in afternoon of fair weather are selected to give birth to twig then as the explant of root culture；
The silt on surface first with the explant of collection is cut to 15cm, is rinsed afterwards in interior with tap water；
First branch is cleaned with detergent, dipping a drop Tween 80 after foams of washing powder is rinsed well with Glass rod carries out depth
Layer cleaning, is placed in explant under flowing water afterwards and rinses 2 hours.
3rd, the sterilizing of explant and clip：
The sterilizing of explant：The explant rinsed well is placed on superclean bench, first 15s is soaked with 75% ethanol,
With aseptic water washing 4 times, then with 0.1% mercuric chloride immersion 3min, afterwards with aseptic water washing 5 times；
The clip of explant after sterilizing：Sterilized explant base portion otch is wiped out, is blotted with sterilized filter paper outer
The moisture on implant surface, is cut into long 1cm length the segment with bud, is inoculated in root media, one explant of per bottle of inoculation.
4th, root culture：
Prescription of rooting medium is：0.2 g/L+ sucrose of 1/2MS basal mediums+IBA, 25 g/L, pH value is 6.0；Culture
Condition is：26 DEG C of temperature, intensity of illumination 2000lx, light application time 10h/d；Sucrose, agar is not contained wherein in MS basal mediums
Concentration is 5g/L.
Embodiment 4
The raw Ramulus et Folium Tamariciss rooting method for tissue culture of sand of the present invention is comprised the following steps that：
1st, the selection of explant：
The selection of suitable explant is the basis of tissue culture, of the invention to select husky raw Ramulus et Folium Tamariciss to give birth to twig then as root culture
Explant, study its tissue culture and take root situation.
The husky raw Ramulus et Folium Tamariciss current-year branch of clip in afternoon of fair weather is selected as the explant of root culture；
The silt on surface first with the explant of collection is cut to 12cm, is rinsed afterwards in interior with tap water；
First branch is cleaned with detergent, dipping a drop Tween 80 after foams of washing powder is rinsed well with Glass rod carries out depth
Layer cleaning, is placed in explant under flowing water afterwards and rinses 4 hours.
3rd, the sterilizing of explant and clip：
The sterilizing of explant：The explant rinsed well is placed on superclean bench, first 12s is soaked with 75% ethanol,
With aseptic water washing 3 times, then with 0.1% mercuric chloride immersion 4min, afterwards with aseptic water washing 6 times；
The clip of explant after sterilizing：Sterilized explant base portion otch is wiped out, is blotted with sterilized filter paper outer
The moisture on implant surface, is cut into long 1.cm length the segment with bud, is inoculated in root media, one explant of per bottle of inoculation
Body.
4th, root culture：
Prescription of rooting medium is：0.2 g/L+ sucrose of 1/2MS basal mediums+IBA, 30 g/L, pH value is 5.9；Culture
Condition is：23 DEG C of temperature, intensity of illumination 1900lx, light application time 11h/d；Sucrose, agar is not contained wherein in MS basal mediums
Concentration is 5g/L.
Finally it should be noted that：The preferred embodiments of the present invention are the foregoing is only, the present invention is not limited to,
Although being described in detail to the present invention with reference to the foregoing embodiments, for a person skilled in the art, which still may be used
To modify to the technical scheme described in foregoing embodiments, or equivalent is carried out to which part technical characteristic.
All any modification, equivalent substitution and improvements within the spirit and principles in the present invention, made etc., should be included in the present invention's
Within protection domain.
Claims (9)
1. the raw Ramulus et Folium Tamariciss rooting method for tissue culture of a planting sand, it is characterised in that：Comprise the following steps：
（1）The selection of explant：Husky raw Ramulus et Folium Tamariciss are selected to give birth to twig then as the explant of root culture；
（2）Explant pre-treatment；
（3）The sterilizing of explant and clip；
（4）Root culture：Root culture culture medium prescription is：1/2MS basal medium+IBA 0.2g/L+ sucrose 15-30
g/L；PH value is 5.8-6.0；Root culture condition of culture is：24 ± 2 DEG C of temperature, intensity of illumination 1500-2000lx, light application time
10-14 h/d；Sucrose is not contained in the MS basal mediums, and agar concentration is 5g/L.
2. method according to claim 1, it is characterised in that：Step（2）Described in explant pre-treatment be select it is sunny
The explant of collection is first cut indoors by the husky raw Ramulus et Folium Tamariciss current-year branch of clip in afternoon of weather as the explant of root culture
To 10-15cm, the silt on surface is rinsed afterwards with tap water；Then branch is cleaned with detergent, foams of washing powder is rinsed into dry
A drop Tween 80 is dipped with Glass rod after net carrying out deep layer and clean, explant is placed in into flushing 2-6 hours under flowing water afterwards.
3. method according to claim 1, it is characterised in that：Step（3）Described in the sterilizing of explant be to rinse dry
Net explant is placed on superclean bench, first with 75% ethanol immersion 10-15s, with aseptic water washing 3-4 time, then with 0.1%
Mercuric chloride immersion 3-6 min, different according to old tender degree, old branch bar soak time is longer, is 5-6 min；When shoot soaks
Between it is short, be 3-4min, afterwards with aseptic water washing 5-7 time.
4. method according to claim 1, it is characterised in that：Step（3）Described in the clip of explant be to have been sterilized
Explant base portion otch wipe out, blot the moisture on explant surface with sterilized filter paper, be cut into long 1-2cm length and with bud
Segment, is inoculated in root media, one explant of per bottle of inoculation.
5. method according to claim 1, it is characterised in that：Step（4）Described in root culture culture medium prescription be：1/
2MS basal medium+IBA 0.2g/L+ sucrose 15-20 g/L.
6. method according to claim 5, it is characterised in that：Step（4）Described in root culture culture medium prescription be：1/
2MS basal medium+IBA 0.2g/L+ sucrose 15g/L.
7. the raw Ramulus et Folium Tamariciss culture medium for tissue culture of a planting sand, it is characterised in that：The formula of the culture medium is：The culture of 1/2MS bases
Base+IBA 0.2g/L+ sucrose 15-30g/L.
8. culture medium according to claim 7, it is characterised in that：The formula of the culture medium is：1/2MS basal mediums
+ IBA 0.2g/L+ sucrose 15-20g/L.
9. culture medium according to claim 8, it is characterised in that：The formula of the culture medium is：1/2MS basal mediums
+ IBA 0.2g/L+ sucrose 15g/L.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107810850A (en) * | 2017-08-22 | 2018-03-20 | 甘肃省治沙研究所 | A kind of populus simonii and diversiform-leaved poplar（Small × diversiform-leaved poplar）The breeding method of cenospecies regrowth |
| CN116034873A (en) * | 2022-12-02 | 2023-05-02 | 武威市林业科学研究院 | Tamarix chinensis tissue rapid propagation method |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104542294A (en) * | 2015-01-14 | 2015-04-29 | 上海杉一植物科技有限公司 | Rooting culture method for tamarix chinensis tissue culture seedlings |
| 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 |
| CN105850740A (en) * | 2016-04-19 | 2016-08-17 | 青岛农业大学 | Screening method for pear homologous transgene functional verification material |
| CN106538395A (en) * | 2017-01-18 | 2017-03-29 | 西藏农牧学院 | A kind of method for improving endangered plants Tibet Paeonia ludlowii planting percent and seedling percent |
-
2016
- 2016-10-27 CN CN201610958954.2A patent/CN106508678B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104542294A (en) * | 2015-01-14 | 2015-04-29 | 上海杉一植物科技有限公司 | Rooting culture method for tamarix chinensis tissue culture seedlings |
| 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 |
| CN105850740A (en) * | 2016-04-19 | 2016-08-17 | 青岛农业大学 | Screening method for pear homologous transgene functional verification material |
| CN106538395A (en) * | 2017-01-18 | 2017-03-29 | 西藏农牧学院 | A kind of method for improving endangered plants Tibet Paeonia ludlowii planting percent and seedling percent |
Non-Patent Citations (1)
| Title |
|---|
| 张锦春等: "沙生柽柳扦插苗生根条件优化研究", 《中国农学通报》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107810850A (en) * | 2017-08-22 | 2018-03-20 | 甘肃省治沙研究所 | A kind of populus simonii and diversiform-leaved poplar（Small × diversiform-leaved poplar）The breeding method of cenospecies regrowth |
| CN107810850B (en) * | 2017-08-22 | 2020-06-16 | 甘肃省治沙研究所 | Cultivation method of regenerated seedlings of hybrid seeds of populus microphylla and populus diversifolia |
| CN116034873A (en) * | 2022-12-02 | 2023-05-02 | 武威市林业科学研究院 | Tamarix chinensis tissue rapid propagation method |
| CN116034873B (en) * | 2022-12-02 | 2023-11-17 | 武威市林业科学研究院 | Tamarix chinensis tissue rapid propagation method |
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