CN106937600B - A kind of Pizhou City's white garlic method for tissue culture and culture medium combination - Google Patents
A kind of Pizhou City's white garlic method for tissue culture and culture medium combination Download PDFInfo
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Abstract
The invention discloses a kind of Pizhou City's white garlic method for tissue culture and culture medium to combine, and is related to technology for garlic tissue culture field.White garlic method for tissue culture in Pizhou City's disclosed by the invention carries out Stem tip induction culture by explant of the stem apex through Pizhou City's white garlic bulb, the blade of formation is subjected to blade Fiber differentiation again, form callus, callus carries out callus differentiation culture and culture of rootage again, obtains complete Pizhou City's white garlic tissue culturing seedling.The method for tissue culture effectively improves stem apex survival rate, reduces the pollution rate in operating process, and growth coefficient with higher is conducive to garlic seedling such as Pizhou City's white garlic the factorial production up to 42.5.
Description
Technical field
The present invention relates to technology for garlic tissue culture fields, in particular to a kind of Pizhou City's white garlic method for tissue culture
And culture medium combination.
Background technique
Garlic is the pillar industry of Pizhou City, Jiangsu Province agricultural, there is more than 2000 years cultivation histories.Pizhou City's white garlic is " a with it
Greatly, color is white, smooth, circle is real, do not dissipate valve, pungent degree is moderate " and enjoy great prestige be at home and abroad local farmers main breed.In recent years
Investigation discovery, Pizhou City's white garlic (Allium sativum L.) disease than more serious, brings very huge economic loss to garlic farmers,
Its reason causes breediness to degenerate mainly due to the long-term vegetative propagation of Pizhou City's white garlic, and resistance is deteriorated, yielding ability and anti-
Sick ability reduces.In vitro callus inductive technology can be effectively prevented garlic cultivar degeneration, improve breeding coefficient, keep and improve and is big
The good strains of seeds of garlic.
Currently, Garlic Tissue culture studies have been reported that both at home and abroad, but mostly directly carried out with explants such as stem apex, blades
Evoked callus, this method for tissue culture is low and phenomena such as blade pollution rate is high there are stem apex survival rate, further results in
The problems such as breeding coefficient is low, reproduction speed is slow.
Summary of the invention
The purpose of the present invention is to provide a kind of Pizhou City's white garlic method for tissue culture, which can not only be mentioned
Pollution rate in high stem apex survival rate, reduction operating process, additionally it is possible to improve the breeding coefficient and detoxification efficiency of Pizhou City's white garlic.
Another object of the present invention is to provide a kind of culture medium combination for Pizhou City's white garlic tissue cultures, the culture mediums
It combines suitable for that can not only improve stem apex survival rate to Pizhou City's white garlic progress tissue cultures, reduce the pollution operating process
Rate, additionally it is possible to improve Pizhou City's white garlic breeding coefficient and reproduction speed.
The present invention is implemented as follows:
A kind of Pizhou City's white garlic method for tissue culture comprising:
It takes the stem apex of Pizhou City's white garlic bulb to be placed in progress Stem tip induction culture on Stem tip induction differential medium, forms leaf
Piece；
Blade is placed in progress blade Fiber differentiation on blade inductive differentiation medium, forms callus；
Callus is subjected to callus differentiation culture and culture of rootage, obtains complete Pizhou City's white garlic tissue culturing seedling.
A kind of culture medium combination for Pizhou City's white garlic tissue cultures comprising at least two in following culture medium:
Stem apex for Pizhou City's white garlic bulb carries out Stem tip induction differentiation culture to form the Stem tip induction differentiation training of blade
Support base；
Blade Fiber differentiation is carried out for the blade to form the blade inductive differentiation medium of callus；
Callus differentiation culture is carried out for the callus to form the callus differential medium of Multiple Buds；
And the root media of culture of rootage is carried out for the Multiple Buds.
Compared with prior art, the beneficial effects of the present invention are:
White garlic method for tissue culture in Pizhou City's provided by the invention, using the stem apex of Pizhou City's white garlic bulb as explant, in stem apex
Stem tip induction culture is carried out on inductive differentiation medium, forms blade, blade is subjected to leaf on blade inductive differentiation medium
Piece Fiber differentiation forms callus；Through differentiation culture and culture of rootage, it is white to obtain complete Pizhou City again for the callus of formation
Garlic tissue culturing seedling.Relative to existing method for tissue culture, the present invention using Fiber differentiation, that is, Stem tip induction culture twice and
Blade Fiber differentiation after Stem tip induction culture effectively improves stem apex survival rate, reduces the pollution in operating process
Rate also improves the breeding coefficient (up to 42.5%) and reproduction speed of Pizhou City's white garlic.White garlic tissue in Pizhou City's provided by the invention
Cultural method has many advantages, such as that breeding coefficient is high, reproduction speed is fast, seedling detoxification efficiency is good；Be conducive to the seedling industrialized life of garlic kind
It produces.
In addition, culture medium provided by the invention combines each culture rank suitable for the tissue culture procedures of Pizhou City's white garlic
Section can not only improve stem apex survival rate, reduce the pollution rate in operating process, additionally it is possible to improve the breeding of Pizhou City's white garlic seedling
Coefficient, reproduction speed and detoxification efficiency.
Detailed description of the invention
In order to illustrate the technical solution of the embodiments of the present invention more clearly, below will be to needed in the embodiment attached
Figure is briefly described, it should be understood that the following drawings illustrates only certain embodiments of the present invention, therefore is not construed as pair
The restriction of range for those of ordinary skill in the art without creative efforts, can also be according to this
A little attached drawings obtain other relevant attached drawings.
Fig. 1 is the photo of the differentiated result for Pizhou City's white garlic bulb stem apex that the embodiment of the present invention 1 provides；
Fig. 2 is the photo for the result that the blade that the embodiment of the present invention 1 provides is induced to differentiate to form callus；
Fig. 3 is the photo of the result for the callus form Multiple Buds that the embodiment of the present invention 1 provides；
Fig. 4 is the photo for complete Pizhou City's white garlic tissue culturing seedling that the embodiment of the present invention 1 provides.
Specific embodiment
It in order to make the object, technical scheme and advantages of the embodiment of the invention clearer, below will be in the embodiment of the present invention
Technical solution be clearly and completely described.The person that is not specified actual conditions in embodiment, according to normal conditions or manufacturer builds
The condition of view carries out.Reagents or instruments used without specified manufacturer is the conventional production that can be obtained by commercially available purchase
Product.
A kind of Pizhou City's white garlic method for tissue culture of the invention and culture medium combination are specifically described below.
On the one hand, the present invention provides a kind of Pizhou City's white garlic method for tissue culture comprising following steps:
S1 aseptic process step:
The above-mentioned Pizhou City's white garlic bulb for choosing full no disease and pests harm, peels off the scale of above-mentioned Pizhou City's white garlic bulb, takes Pizhou City
The stem apex of white garlic bulb, is placed in flowing water and rinses 10-12min, impregnates 30-35s with the alcohol of 70-80%, then gone out with thimerosal
Bacterium 15-20min, with aseptic water washing 3-5 times.
S2 Stem tip induction incubation step:
The stem apex of Pizhou City's white garlic bulb after aseptic process of learning from else's experience is placed in progress stem apex on Stem tip induction differential medium and lures
Culture is led, Pizhou City's white garlic seedling is formed, takes the blade of Pizhou City's white garlic seedling for subsequent step.
Further, the length of stem apex is 1.0~1.5cm.
Wherein, the ingredient of Stem tip induction differential medium are as follows: MS+1~2mg/L 6-BA+0.18~0.22mg/L NAA+
6.8~7.2g/L agar+28~32g/L sucrose, pH value 5.8.
Wherein, the condition of Stem tip induction culture are as follows: 23-27 DEG C of temperature, intensity of illumination 1200-1500lx, illumination 11-13h/
d.Incubation time is 40d or so.
Above-mentioned Stem tip induction differential medium is conducive to the stem apex differentiation of Pizhou City's white garlic bulb, and the differentiation rate of stem apex is reachable
60% or more, culture 42d can differentiate Pizhou City's white garlic seedling, and plant height is up to 6.8cm.
S3 blade Fiber differentiation step:
Above-mentioned blade is placed in progress blade Fiber differentiation on blade inductive differentiation medium, forms callus.
Wherein, the ingredient of blade inductive differentiation medium are as follows: MS+1~2mg/L 6-BA+0.18~0.22mg/L NAA+
6.8~7.2g/L agar+28~32g/L sucrose, pH value 5.8.
Wherein, the condition of blade Fiber differentiation are as follows: 23-27 DEG C of temperature, intensity of illumination 1200-1500lx, illumination 11-13h/
d.Incubation time is 60d or so.
S4 callus breaks up incubation step:
After above-mentioned callus is cut into the block of 0.5cm or so, it is inoculated in callus differential medium and cultivates, grown thickly
Bud.
Wherein, the ingredient of callus differential medium are as follows: MS+0.4~0.8mg/L 6-BA+0.08~0.12mg/L NAA+
6.8~7.2g/L agar+28~32g/L sucrose, pH value 5.8.
Wherein, the condition of callus differentiation culture are as follows: 23-27 DEG C of temperature, intensity of illumination 1200-1500lx, illumination 11-13h/
d。
Above-mentioned callus differential medium is conducive to the differentiation of callus, improves differentiation rate, culture 30d or so differentiation rate
Up to 69.2%, the period is short, average bud number 2.55, and average height 4.8cm continues to cultivate 60d or so, and growth coefficient reaches
42.5, the form of Multiple Buds is also relatively thick, is conducive to late growth.
S5 culture of rootage step:
Will be above 3cm, the Multiple Buds of growing way stalwartness are cut, be placed in root media and cultivate, so that Multiple Buds send out roots
System, culture 60d or so, forms complete Pizhou City's white garlic tissue culturing seedling.
Wherein, the ingredient of root media are as follows: MS+0.48~0.52mg/L 6-BA+0.8~1.1mg/L NAA+0.018
~0.022% active carbon+6.8~7.2g/L agar+28~32g/L sucrose, pH value 5.8.
Wherein, the condition of culture of rootage are as follows: 23-27 DEG C of temperature, intensity of illumination 1200-1500lx, illumination 11-13h/d.
White garlic method for tissue culture in Pizhou City's provided by the invention, using the stem apex of Pizhou City's white garlic bulb as explant, through sterile
After processing, Stem tip induction culture is carried out on Stem tip induction differential medium, forms blade, and blade is induced into differentiation training in blade
It supports and carries out blade Fiber differentiation on base, form callus；The callus of formation through differentiation culture and culture of rootage, obtains again
White garlic tissue culturing seedling, complete Pizhou City.Relative to existing method for tissue culture, the present invention is using Fiber differentiation, that is, stem twice
Sharp Fiber differentiation and the blade Fiber differentiation after Stem tip induction culture effectively improve stem apex survival rate, reduce operation
Pollution rate in the process also improves breeding coefficient (up to 42.5), reproduction speed and the detoxification efficiency of Pizhou City's white garlic.
In short, white garlic method for tissue culture in Pizhou City's provided by the invention has, breeding coefficient is high, reproduction speed is fast, seedling is de-
The advantages that toxic effect fruit is good.
On the other hand, the present invention provides a kind of culture medium combinations for Pizhou City's white garlic tissue cultures comprising as follows
At least two in culture medium:
Stem apex for Pizhou City's white garlic bulb carries out Stem tip induction differentiation culture to form the Stem tip induction differentiation training of blade
Support base；
Blade Fiber differentiation is carried out for the blade to form the blade inductive differentiation medium of callus；
Callus differentiation culture is carried out for the callus to form the callus differential medium of Multiple Buds；
And the root media of culture of rootage is carried out for the Multiple Buds.
Further, the ingredient of above-mentioned Stem tip induction differential medium and above-mentioned blade inductive differentiation medium is equal are as follows: MS+
1~2mg/L 6-BA+0.18~0.22mg/L NAA+6.8~7.2g/L agar+28~32g/L sucrose, pH value 5.8；
The ingredient of above-mentioned callus differential medium are as follows: MS+0.4~0.8mg/L 6-BA+0.08~0.12mg/L NAA+
6.8~7.2g/L agar+28~32g/L sucrose；
The ingredient of above-mentioned root media are as follows: MS+0.48~0.52mg/L 6-BA+0.8~1.1mg/L NAA+0.018
~0.022% active carbon+6.8~7.2g/L agar+28~32g/L sucrose.
Culture medium provided by the invention combines each cultivation stage suitable for the tissue culture procedures of Pizhou City's white garlic:
Cultivation stage is broken up in the induction that Stem tip induction differential medium is suitable for stem apex, and stem apex is made to be differentiated to form Pizhou City's white garlic
Seedling improves the differentiation rate (61% or more) and survival rate of stem apex.
The ingredient of blade inductive differentiation medium is consistent with Stem tip induction differential medium, suitable for above-mentioned formation
The blade of Pizhou City's white garlic seedling carries out blade induction differentiation culture, and blade is made to be differentiated to form callus.
The callus that callus differential medium is suitable for callus breaks up cultivation stage, is differentiated to form callus largely
Multiple Buds, improve callus differentiation rate (61% or more) and budding number.
Root media is suitable for the culture of rootage stage of Multiple Buds, so that Multiple Buds is taken root, forms complete Pizhou City's white garlic
Tissue culturing seedling.
Above-mentioned culture medium combination is carried out for the nutritional need of each cultivation stage in the tissue culture procedures of Pizhou City's white garlic
Specifically it is arranged, improves stem apex survival rate, breeding coefficient and reproduction speed on the whole, be effectively reduced in operating process
Pollution rate.
Feature and performance of the invention are described in further detail with reference to embodiments.
Embodiment 1
White garlic method for tissue culture in Pizhou City's provided in this embodiment, includes the following steps:
1.1 aseptic process steps:
Pizhou City's white garlic (the Allium sativum L.) bulb for choosing full no disease and pests harm, peels off Pizhou City's white garlic bulb
Scale takes the stem apex of Pizhou City's white garlic bulb, is placed in flowing water and rinses 10min or so, is placed on superclean bench, with 75% wine
Essence impregnate 30s, then with thimerosal (84 thimerosal) sterilizing 15min, with aseptic water washing 5 times.
1.2 Stem tip induction incubation steps:
Stem apex that take 1-1.5cm size, Pizhou City's white garlic bulb after aseptic process is inoculated in Stem tip induction differentiation training
It supports and carries out Stem tip induction culture on base to form blade.
The condition of Stem tip induction culture are as follows: 25 DEG C of temperature, intensity of illumination 1200-1500lx, illumination 12h/d.Incubation time
For 40d or so.
Wherein, the ingredient of Stem tip induction differential medium are as follows: MS+2mg/L 6-BA+0.2mg/L NAA+7g/L agar+
30g/L sucrose, pH5.8.
It observes stem apex and breaks up situation, as a result as shown in Figure 1, culture 42d stem apex can differentiate Pizhou City's white garlic seedling.
And using ingredient be MS+0.5mg/L 6-BA+0.2mg/L NAA+7g/L agar+30g/L sucrose (as control 1
Group), MS+2.0mg/L 2,4-D+0mg/L NAA+7g/L agar+30g/L sucrose (as compareing 2 groups), MS+0.5mg/L TDZ
Three kinds of stem apexs containing hormon type and concentration of+0mg/L NAA+7g/L agar+30g/L sucrose (as compareing 3 groups)
Inductive differentiation medium detects the indexs such as differentiation rate and the plant height of stem apex, the results are shown in Table 1 as control.
The influence that 1 hormon concentration of table and hormone kind break up stem apex
Table 1 the results show that the ingredient of stem apex in the present embodiment be MS+2mg/L 6-BA+0.2mg/L NAA+7g/L
Agar+30g/L sucrose, the differentiation rate on the Stem tip induction differential medium of pH5.8 is up to 61.5%, hence it is evident that be higher than 1 group of control,
2 groups and 3 groups of control are compareed, average plant height reaches 6.8cm, and plant growing way is thick.Illustrate Stem tip induction provided in an embodiment of the present invention
Differential medium effectively improves the differentiation rate and survival rate of stem apex.
1.3 blade Fiber differentiation steps:
The blade that above-mentioned steps obtain Pizhou City's white garlic seedling is placed on blade inductive differentiation medium and carries out blade induction training
It supports, to form callus.
Wherein, the ingredient of blade inductive differentiation medium are as follows: MS+2mg/L 6-BA+0.2mg/L NAA+7g/L agar+
30g/L sucrose, pH5.8.
Wherein, the condition of blade Fiber differentiation are as follows: 25 DEG C of temperature, intensity of illumination 1200-1500lx, illumination 12h/d.Culture
Time is 60d or so, as a result as shown in Fig. 2, blade differentiates a large amount of callus lines.
1.4 callus break up incubation step:
The callus that 60d or so is formed will be cultivated to be cut into small pieces after (0.5cm or so size), be inoculated in callus differentiation training
It supports in base and carries out Multiplying culture, obtain Multiple Buds.As a result as shown in figure 3, callus islands hyperplasia goes out a large amount of Multiple Buds.
Wherein, the ingredient of callus differential medium are as follows: MS+0.8mg/L 6-BA+0.1mg/L NAA+7g/L agar+30g/
L sucrose, pH5.8.
Wherein, the condition of callus differentiation culture are as follows: 25 DEG C of temperature, intensity of illumination 1200-1500lx, illumination 12h/d.
And with the 6-BA containing concentration 0.2mg/L (1 group of control), 1.0mg/L (2 groups of control), 2.0mg/L (3 groups of control)
Three kinds of callus differential mediums (remaining ingredient of control group culture medium and the callus differential medium of the present embodiment are identical) make
For control, differentiation rate (the bottle number of differentiated callus accounts for the percentage for being inoculated with total bottle number), (every bottle of number of the budding of callus are counted
The bud number of generation) and average height, the results are shown in Table 2.
Influence of the 2 hormon concentration of table to the induction differentiation of blade callus
Table 2 the results show that callus tissue culture 30d in the present embodiment or so differentiation rate up to 69.2%, average bud number
2.55, average height 4.8cm, the form of obtained Multiple Buds is also relatively thick, and indices data are above 1 group of control, control 2
3 groups of group and control；Continue to cultivate 60d or so, the growth coefficient of the present embodiment (is expanded by the bulb stem apex of single Pizhou City's white garlic
Tissue-cultured seedling quantity) up to 42.5.Thus illustrate that callus differential medium provided in an embodiment of the present invention effectively improves more
The differentiation rate and budding number of injured tissue.
1.5 culture of rootage steps:
Will be above 3cm, the Multiple Buds of growing way stalwartness are cut, be placed in root media and cultivate, so that Multiple Buds send out roots
System, culture 60d or so, obtains complete Pizhou City's white garlic tissue culturing seedling (as shown in Figure 4).
Wherein, the ingredient of root media are as follows: MS+0.5mg/L 6-BA+1.0mg/L NAA+0.02% active carbon+7.g/
L agar+30g/L sucrose, pH5.8.
Wherein, the condition of culture of rootage are as follows: 25 DEG C of temperature, intensity of illumination 1200-1500lx, illumination 12h/d.
Embodiment 2
White garlic method for tissue culture in Pizhou City's provided in this embodiment, includes the following steps:
2.1 aseptic process steps:
With step 1.1
2.2 Stem tip induction incubation steps:
It is essentially identical with step 1.2, unlike: the ingredient of Stem tip induction differential medium are as follows: MS+1mg/L 6-BA+
0.2mg/L NAA+7g/L agar+30g/L sucrose, pH5.8.
It should be noted that in other examples, the concentration of the 6-BA in Stem tip induction differential medium can be
1.2,1.4,1.6, any one in 1.8mg/L, as long as in the range of 1-2mg/L；The concentration of NAA can be
0.18,0.19,0.21, any one in 0.22mg/L, if in the range of 0.18~0.22mg/L, agar it is dense
Degree can be 6.8,6.9,7.1, any one in 7.2g/L, as long as within the scope of 6.8~7.2g/L；The concentration of sucrose
It can be 28,29,31, any one in 32g/L, as long as within the scope of 28~32g/L.
It observes stem apex and breaks up situation, and count differentiation rate and plant height.The results are shown in Table 1.
In the present embodiment, Shoot Tip Culture 42d or so differentiates Pizhou City's white garlic seedling, and the differentiation rate of stem apex is average up to 62.5%
Plant height reaches 4cm, and plant growing way is thin.
2.3 blade Fiber differentiation steps:
It is essentially identical with step 1.3, unlike: the ingredient of blade inductive differentiation medium are as follows: MS+1mg/L 6-BA+
0.2mg/L NAA+7g/L agar+30g/L sucrose, pH5.8.
It should be noted that in other examples, the concentration of the 6-BA in blade inductive differentiation medium can be
1.2,1.4,1.6, any one in 1.8mg/L, as long as in the range of 1-2mg/L；The concentration of NAA can be
0.18,0.19,0.21, any one in 0.22mg/L, if in the range of 0.18~0.22mg/L, agar it is dense
Degree can be 6.8,6.9,7.1, any one in 7.2g/L, as long as within the scope of 6.8~7.2g/L；The concentration of sucrose
It can be 28,29,31, any one in 32g/L, as long as within the scope of 28~32g/L.
2.4 callus break up incubation step:
It is essentially identical with step 1.4, unlike:
The ingredient of callus differential medium are as follows: MS+0.4mg/L 6-BA+0.1mg/L NAA+7g/L agar+30g/L sugarcane
Sugar, pH5.8.
It should be noted that in other examples, the concentration of the 6-BA in callus differential medium can be 0.45,
0.5,0.6, any one in 0.75mg/L, as long as in the range of 0.4~0.8mg/L；The concentration of NAA can be
0.08,0.09,0.11, any one in 0.12mg/L, if in the range of 0.08~0.12mg/L, agar it is dense
Degree can be 6.8,6.9,7.1, any one in 7.2g/L, as long as within the scope of 6.8~7.2g/L；The concentration of sucrose
It can be 28,29,31, any one in 32g/L, as long as within the scope of 28~32g/L.
Differentiation rate, budding number and the average height of callus are observed and count, the results are shown in Table 2.
In the present embodiment, 30d is cultivated, the differentiation rate of callus is up to 61.1%, averagely bud number 1.91, average height
Form for 3.8cm, a small amount of cultivating seedling is thicker.
2.5 culture of rootage steps:
With step 1.5.
It should be noted that in other examples, the concentration of the 6-BA in root media can be 0.48,
0.49,0.51, any one in 0.52mg/L, as long as in the range of 0.48~0.52mg/L；The concentration of NAA can be with
It is 0.8,0.9, any one in 1.1mg/L, as long as in the range of 0.8~0.11mg/L；The concentration of active carbon can
To be any one in 0.018,0.019,0.021,0.022%, as long as in 0.018~0.022% range；Fine jade
The concentration of rouge can be 6.8,6.9,7.1, any one in 7.2g/L, as long as within the scope of 6.8~7.2g/L；Sucrose
Concentration can be 28,29,31, any one in 32g/L, as long as within the scope of 28~32g/L.
It should also be noted that, in other examples, Stem tip induction culture, blade Fiber differentiation, callus differentiation training
Support and the condition of culture of culture of rootage can be following situation: temperature can be any one in 23,24,26,27 DEG C, as long as
It is in the range of 23-27 DEG C；As long as intensity of illumination can be 1200,1300,1400, in 1500lx any one,
In the range of 1200-1500lx；Illumination can be any one in 11,12,13 (h/d), as long as daily in illumination
In the range of 11-13h.
To sum up, white garlic method for tissue culture in Pizhou City's provided by the invention, using the stem apex of Pizhou City's white garlic bulb as explant, warp
After aseptic process, Stem tip induction culture is carried out on Stem tip induction differential medium, forms blade, by blade in blade induction point
Change and carry out blade Fiber differentiation on culture medium, forms callus；The callus of formation is cultivated again through differentiation and culture of rootage,
Obtain complete Pizhou City's white garlic tissue culturing seedling.Relative to existing method for tissue culture, the present invention is using Fiber differentiation twice
That is Stem tip induction culture and the blade Fiber differentiation after Stem tip induction culture effectively improves stem apex survival rate, reduces
Pollution rate in operating process also improves the breeding coefficient (up to 42.5) and reproduction speed of Pizhou City's white garlic, is conducive to garlic
The factorial production of seedling especially Pizhou City's white garlic.
In short, white garlic method for tissue culture in Pizhou City's provided by the invention has, breeding coefficient is high, reproduction speed is fast, seedling is de-
The advantages that toxic effect fruit is good.
The foregoing is only a preferred embodiment of the present invention, is not intended to restrict the invention, for the skill of this field
For art personnel, the invention may be variously modified and varied.All within the spirits and principles of the present invention, made any to repair
Change, equivalent replacement, improvement etc., should all be included in the protection scope of the present invention.
Claims (4)
1. a kind of Pizhou City's white garlic method for tissue culture, characterized in that it comprises:
It takes the stem apex of Pizhou City's white garlic bulb to be placed in progress Stem tip induction culture on Stem tip induction differential medium, forms blade；
The blade is placed in progress blade Fiber differentiation on blade inductive differentiation medium, forms callus；
The callus is subjected to callus differentiation culture and culture of rootage, obtains complete Pizhou City's white garlic tissue culturing seedling；Institute
The ingredient for stating Stem tip induction differential medium and the blade inductive differentiation medium is equal are as follows: MS+1~2mg/L 6-BA+0.18
~0.22mg/L NAA+6.8~7.2g/L agar+28~32g/L sucrose；
The callus differentiation culture includes: after the callus is cut into block, to be inoculated in callus differential medium and cultivate, obtain
To Multiple Buds；
The ingredient of the callus differential medium are as follows: MS+0.4~0.8mg/L 6-BA+0.08~0.12mg/L NAA+6.8~
7.2g/L agar+28~32g/L sucrose；
The culture of rootage includes: that will be above the Multiple Buds of 3cm to be placed in root media and cultivated；
The ingredient of the root media are as follows: MS+0.48~0.52mg/L 6-BA+0.8~1.1mg/L NAA+0.018~
0.022% active carbon+6.8~7.2g/L agar+28~32g/L sucrose.
2. white garlic method for tissue culture in Pizhou City's according to claim 1, which is characterized in that the Stem tip induction culture, institute
State the condition of culture of blade Fiber differentiation, the callus differentiation culture and the culture of rootage are as follows: 23-27 DEG C of temperature, illumination are strong
Spend 1200-1500lx, illumination 11-13h/d.
3. white garlic method for tissue culture in Pizhou City's described in any one of -2 according to claim 1, which is characterized in that described in progress
It further include aseptic process before Stem tip induction culture, the aseptic process includes: to take the stem apex, is placed in flowing water and rinses 10-
12min impregnates 30-35s with the alcohol of 70-80%, then is sterilized 15-20min with thimerosal, with aseptic water washing 3-5 times.
4. a kind of culture medium for Pizhou City's white garlic tissue cultures combines, which is characterized in that it includes following culture medium:
Stem apex for Pizhou City's white garlic bulb carries out Stem tip induction differentiation culture to form the Stem tip induction differential medium of blade；
Blade Fiber differentiation is carried out for the blade to form the blade inductive differentiation medium of callus；
Callus differentiation culture is carried out for the callus to form the callus differential medium of Multiple Buds；
And the root media of culture of rootage is carried out for the Multiple Buds；
The ingredient of the Stem tip induction differential medium and the blade inductive differentiation medium is equal are as follows: MS+1~2mg/L 6-BA
+ 0.18~0.22mg/L NAA+6.8~7.2g/L agar+28~32g/L sucrose；
The ingredient of the callus differential medium are as follows: MS+0.4~0.8mg/L 6-BA+0.08~0.12mg/L NAA+6.8~
7.2g/L agar+28~32g/L sucrose；
The ingredient of the root media are as follows: MS+0.48~0.52mg/L 6-BA+0.8~1.1mg/L NAA+0.018~
0.022% active carbon+6.8~7.2g/L agar+28~32g/L sucrose.
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