CN104472359B - A kind of ginseng adventitious root proliferative induction method - Google Patents
A kind of ginseng adventitious root proliferative induction method Download PDFInfo
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
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Abstract
The present invention relates to a kind of ginseng adventitious root proliferative induction method, comprises the following steps：Induced synthesis adventitious root in solid inducing culture is inoculated in after Radix Ginseng tissue cultured seedling is cut into tissue fritter, and being inoculated in after the adventitious root is cut into adventitious root fritter in liquid proliferated culture medium carries out the enrichment culture of adventitious root；Wherein, the solid inducing culture and liquid proliferated culture medium are to contain the indolebutyric acid that concentration is 1 10mg/L with 1/2MS (N) culture medium as minimal medium.Using method provided by the present invention, the adventitious root quantity that induction is obtained can be significantly improved, and, the adventitious root that induction is obtained is carried out after enrichment culture, the adventitious root rate of increase is higher compared with the method for root induction is carried out using calluss, and cultivation effect is more preferable.
Description
Technical field
The present invention relates to field of plant tissue culture technique, is specifically related to a kind of ginseng adventitious root proliferative induction method.
Background technology
Radix Ginseng (Panax ginseng C.A.Meyer) is the perennial dicotyledon of Araliaceae Panax, be ancient and
Famous and precious Chinese crude drug, with multiple effects such as regulation body immunity, anticancer and anti-diabetics.Ginsenoside is its main work
Property material.In recent years, both at home and abroad for the demand rapid growth of Radix Ginseng.But, excessively excavated due to long-term, Radix Ginseng wild resource
Substantially exhausted, and the culture of ginseng cycle is long, the impact of easy climate, pest and disease damage and cultivation condition, it is difficult to meet increasing
The market demand.Ginseng tissue culture technique cycle is short, is not subject to seasonal restrictions, and easily carries out large-scale industrial production, tool
There is very big development prospect.
Induce the hairy root growth that Radix Ginseng is produced rapid using Agrobacterium rhizogenes, the Radix Ginseng phase of saponin species and field cultivation
With content is higher than common cell culture, and the system using bioreactor large-scale culture ginseng poultry has also been set up
[Jeong GT,Park DH,Hwang B,et al.Comparison of growth characteristics of Panax
ginseng hairy roots in various bioreactors[J].Appl Biochem Biotechnol,2003,
107:493].However, there is potential safety issue due to the importing of exogenous gene during induction of hairy roots, the therefore technology
Not used for commercialization.
In recent years, the research of ginseng adventitious root is directly induced to achieve greater advance without Agrobacterium rhizogenes conversion.But
Adventitious root is usually to be induced [Sivakumar G, Yu KW, the Paek KY.Production of for producing by ginseng callus
biomass and ginsenosides from adventitious roots of Panax ginseng in
bioreactor cultures[J].Eng Life Sci,2005,4:333].The defect of said method is：Need first to induce
Calluss, cause experimental period long.
Therefore, it is necessary to provide a kind of directly breeding as the root induction of explant for simplicity by the use of Radix Ginseng tissue cultured seedling
Method.
Content of the invention
It is an object of the invention to provide a kind of root of utilization Radix Ginseng tissue cultured seedling, stem section or blade inducing adventitious root produce and
The method that enrichment culture is carried out to adventitious root.
For reaching object above, the invention provides a kind of root induction enrichment procedure, comprises the following steps：
Induced synthesis adventitious root in solid inducing culture is inoculated in after Radix Ginseng tissue cultured seedling is cut into tissue fritter, by institute
State and after adventitious root cuts into adventitious root fritter, be inoculated in liquid proliferated culture medium the enrichment culture for carrying out adventitious root；
Wherein, the solid inducing culture and liquid proliferated culture medium are with 1/2MS (- N) culture medium as basic culture
Base contains the indolebutyric acid (IBA) that concentration is 1-10mg/L.
Preferably in order to obtain more preferable ginseng adventitious root proliferative induction effect, the solid inducing culture and liquid increase
It is to contain indolebutyric acid of the concentration as 2-5mg/L with 1/2MS (- N) culture medium as minimal medium to grow culture medium.
Optionally, the solid inducing culture and liquid proliferated culture medium are 1.8-2.2 mass % also containing concentration
Sucrose.
Optionally, the proliferative induction method is comprised the following steps：
1) Radix Ginseng tissue cultured seedling is cut into tissue fritter, is 23-25 DEG C using solid inducing culture in cultivation temperature
Under dark condition, inducing culture 3-6 weeks obtain adventitious root；
2) by step 1) adventitious root that obtains cuts into adventitious root fritter, using liquid proliferated culture medium in cultivation temperature be
With the speed concussion and cultivate 3-9 week of 50-80rpm under 23-25 DEG C of dark condition.
Optionally, the tissue fritter is to carry out the Radix Ginseng tissue cultured seedling root fritter, right for cutting acquisition to Radix Ginseng tissue cultured seedling root
Radix Ginseng tissue culture seeding stem segment carries out cutting the Radix Ginseng tissue cultured seedling stem fritter of acquisition or Radix Ginseng tissue culture seedling leaf is carried out cutting acquisition
Radix Ginseng tissue cultured seedling leaf fritter.
Optionally, the Radix Ginseng tissue cultured seedling is through seed sprouting or the tissue cultured seedling of explant isolated culture acquisition, tissue culture
The age in days of Seedling is 28-32 days.
Optionally, the kind of the Radix Ginseng tissue cultured seedling is that Radix Ginseng or Da-maya are joined.
Using method provided by the present invention, the adventitious root quantity that induction is obtained can be significantly improved, and, induction is obtained
Adventitious root carry out enrichment culture after, the adventitious root rate of increase is more compared with the method for root induction is carried out using calluss
Height, cultivation effect are more preferable.In addition method provided by the present invention can greatly shorten the time for obtaining ginseng adventitious root, by straight
Is connect Radix Ginseng tissue cultured seedling is carried out root induction propagation avoid by tissue cultured seedling be trained after whole plant obtain explant carry out
The step of induction of callus, shorten experimental period.It is to have been established well using ginseng adventitious root production ginsenoside
Basis, also provide important material for carrying out the basic research of metabolic pathway using ginseng adventitious root.
Description of the drawings
Fig. 1 is the adventitious root that Radix Ginseng tissue cultured seedling different tissues proliferative induction is produced；
In figure, (A) it is situation using Radix Ginseng tissue cultured seedling root induction culture adventitious root；(B) it is using Radix Ginseng tissue cultured seedling stem
The situation of section inducing culture adventitious root；(C) it is situation using Radix Ginseng tissue culture seedling leaf inducing culture adventitious root.
Fig. 2 is that Radix Ginseng tissue cultured seedling different tissues induce the proliferative conditions for producing adventitious root；
In figure, (A) it is tissue cultured seedling root；(B) it is tissue culture seeding stem segment；(C) it is tissue culture seedling leaf；1/2MS(-N)+5mg/L
IBA and 1/2MS (- N)+2mg/L IBA represent type of culture medium；1st represents first time subculture, and 2nd represents second subculture,
3rd represents third time subculture.
Fig. 3 is the enrichment culture that the root-derived callus induction of 5 years stranger's ginsengs produces adventitious root；
1/2MS (- N)+5mg/L IBA and 1/2MS (- N)+2mg/L IBA represent type of culture medium；1st is represented for the first time
Subculture, 2nd represent second subculture, and 3rd represents third time subculture.
Specific embodiment
Below will the present invention is described in detail by specific embodiment.
The invention provides a kind of ginseng adventitious root proliferative induction method, comprises the following steps：
Induced synthesis adventitious root in solid inducing culture is inoculated in after Radix Ginseng tissue cultured seedling is cut into tissue fritter, by institute
State and after adventitious root cuts into adventitious root fritter, be inoculated in liquid proliferated culture medium the enrichment culture for carrying out adventitious root；
Wherein, the solid inducing culture and liquid proliferated culture medium are with 1/2MS (- N) culture medium as basic culture
Base contains the indolebutyric acid that concentration is 1-10mg/L.
In method provided by the present invention, in order to improve induction and propagation effect, the solid inducing culture and
Liquid proliferated culture medium is to contain indolebutyric acid of the concentration as 2-5mg/L with 1/2MS (- N) culture medium as minimal medium.
In the present invention, the Radix Ginseng tissue cultured seedling is to sprout the tissue cultured seedling that obtains through seed or trained through explant in vitro
Support the regeneration plant tissue cultured seedling for obtaining.In the case of preferred, when the method provided using the present invention is to obtaining through isolated culture
Radix Ginseng tissue cultured seedling when carrying out proliferative induction, be obtained in that excellent root induction cultivation effect.
Wherein, method provided by the present invention is for the preparation method of the tissue cultured seedling obtained through seed sprouting is without special
Limit, can obtain according to the conventional processing method in this area, for example, in one embodiment of the invention, through planting
Son sprouting obtains the method for tissue cultured seedling：Learn from else's experience the Radix Ginseng breach seed after Stratificated treatment, peel off shell, through 70% ethanol
After immersion 1 minute, sterilize 20 minutes in 1% sodium hypochlorite, after sterile water wash 3-4 time, complete embryo is stripped out in super-clean bench, connect
Plant in 1/2MS culture medium, after 24 ± 2 DEG C of dark culturing 3 days, proceed to the induction that can be used for adventitious root after 4 weeks are cultivated under light.
Wherein, method provided by the present invention is for by explant culture in vitro system acquisition regeneration plant tissue cultured seedling
Method has no particular limits, and can obtain according to the conventional processing method in this area, for example, real in one kind of the present invention
Apply in mode, the method for obtaining regeneration plant tissue cultured seedling by explant culture in vitro system can be：Ginseng seed is shelled
Afterwards, after with 70% ethanol surface sterilization 1 minute, 1% liquor natrii hypochloritises are proceeded to and is sterilized 20 minutes, sterile water wash 3-4 time, so
Embryo is taken out carefully in superclean bench sterile scalpel afterwards, be seeded to 1/2MS culture medium (containing 2% sucrose and 7g/L agar, pH
Be worth MS+1mg/L2, the culture medium of 4- dichlorphenoxyacetic acids is accessed for 5.8) after 24 ± 2 DEG C of dark culturing 3 days, cutting cotyledon
(containing 3% sucrose and 7g/L agar, pH value is 5.8), in 24 ± 2 DEG C, cultivates under 16 hours illumination/8 hour dark conditions.Take life
(containing 5% sucrose and 10g/L agar, pH value is the MS culture medium that the good embryo callus of long status proceed to without hormone
5.8) induction of somatic embryo is carried out.After about one month, the somatic embryo for inducing is proceeded to the 1/2MS containing 5mg/L gibberellins
(containing 2% sucrose and 2.5g/L plant gels, pH value is the sprouting for 5.8) carrying out plant to culture medium, then proceeds to regeneration plant
NH is not contained4NO31/2MS culture medium (containing 2% sucrose and 2.5g/L plant gels, pH value is 5.8) to carry out root culture, about 3 weeks
Left and right can be taken root, and obtain regeneration plant tissue cultured seedling.
In method provided by the present invention, the tissue fritter is Radix Ginseng tissue cultured seedling root fritter, Radix Ginseng tissue cultured seedling stem is little
Block or Radix Ginseng tissue cultured seedling leaf fritter.
The present invention a kind of preferred embodiment in, described tissue fritter size may be about 5mm × 5mm ×
3mm.Apparatus sterilizing is should be noted that during cutting to the tissue cultured seedling and adventitious root and keeps gnotobasiss to avoid dirt
Dye.
In method provided by the present invention, 1/2MS (- N) culture medium halves the constant MS of other compositions for a great number of elements
(- N) culture medium, wherein, MS (- N) culture medium is free from the MS culture medium of ammonium nitrate.
Wherein, the formula of MS (- N) culture medium is：Potassium nitrate 1900mg/L, potassium dihydrogen phosphate 170mg/L, magnesium sulfate
370mg/L, calcium chloride 440mg/L, potassium iodide 0.83mg/L, boric acid 6.2mg/L, manganese sulfate 22.3mg/L, zinc sulfate 8.6mg/
L, sodium molybdate 0.25mg/L, copper sulfate 0.025mg/L, cobaltous chloride 0.025mg/L, disodiumedetate 37.3mg/L, sulfur
Sour ferrous iron 27.8mg/L, inositol 100mg/L, glycine 2mg/L, thiamine hydrochloride 0.1mg/L, pyridoxine hydrochloride 0.5mg/L, cigarette
Sour 0.5mg/L.
In one embodiment of the invention, in order that Radix Ginseng tissue cultured seedling tissue fritter and induction obtain indefinite
Root obtain more fully nutrition, in the solid inducing culture also containing concentration for 1.8-2.2 mass % sucrose and
Agar of the concentration for 0.65-0.75 mass %.Containing the sugarcane that concentration is 1.8-2.2 mass % in the liquid proliferated culture medium
Sugar.
In one embodiment of the invention, the solid inducing culture, the pH of liquid proliferated culture medium are 5.8-
6.0.
Specifically, the induction and propagation the step of include：
1) Radix Ginseng tissue cultured seedling is cut into tissue fritter, is 23-25 DEG C using solid inducing culture in cultivation temperature
Under dark condition, inducing culture 3-6 weeks obtain adventitious root；
2) by step 1) adventitious root that obtains cuts into adventitious root fritter, using liquid proliferated culture medium in cultivation temperature be
With the speed concussion and cultivate 3-9 week of 50-80rpm under 23-25 DEG C of dark condition.
Wherein in step 1) in, tissue fritter starts to grow adventitious root after cultivating 14-28 days in solid inducing culture.
General, culture in solid inducing culture induces the number of the adventitious root of acquisition reach 4-13 after terminating；Specifically
, induction is generally possible to using Radix Ginseng tissue cultured seedling root fritter obtain 10-13 adventitious root；General using Radix Ginseng tissue cultured seedling stem fritter
8-12 adventitious root of acquisition can be induced；Induction is generally possible to using Radix Ginseng tissue cultured seedling leaf fritter and obtains 4-8 adventitious root.
During the inducing culture and enrichment culture, generally require every 3 time-of-week and change a solid induction training
Foster base or liquid proliferated culture medium carry out successive transfer culture, and the induction of statistical observation adventitious root and propagation feelings in each subculture
Condition.
The preferred embodiment of the present invention is described in detail below by embodiment.It will be appreciated that following implement
Being given merely to play descriptive purpose for example, is not used to limit the scope of the present invention.The technology of this area
Personnel can carry out various modifications and replacement to the present invention in the case of without departing substantially from spirit of the invention and spirit.
In following examples, using the Radix Ginseng kind in Kuandian county.In following examples, Radix Ginseng tissue cultured seedling is by planting
The mode that son is sprouted is obtained, specially：Learn from else's experience the Radix Ginseng breach seed after Stratificated treatment, peel off shell, soak through 70% ethanol
Bubble is sterilized 20 minutes after 1 minute in 1% sodium hypochlorite, after sterile water wash 3-4 time, strips out complete embryo in super-clean bench, inoculation
In 1/2MS culture medium, 24 ± 2 DEG C of dark culturing were proceeded to after 3 days, obtained the tissue cultured seedling that age in days is 30 days.
Root induction rate=(producing the explant number/inoculation explant sum of adventitious root) × 100%.
The adventitious root rate of increase=(propagation harvests the indefinite root fresh weight of indefinite root fresh weight-inoculation on the 3rd week)/inoculation is or not 3rd week
Determine root fresh weight × 100%；
The adventitious root rate of increase=(it is fresh that propagation harvests the 3rd week results adventitious root of indefinite root fresh weight-propagation on the 6th week within 6th week
Again)/propagation the 3rd week harvests indefinite root fresh weight × 100%；
The adventitious root rate of increase=(it is fresh that propagation harvests the 6th week results adventitious root of indefinite root fresh weight-propagation on the 9th week within 9th week
Again)/propagation the 6th week harvests indefinite root fresh weight × 100%；
Embodiment 1
The present embodiment is used for explanation and utilizes Radix Ginseng tissue cultured seedling root induction adventitious root.
Take the root of Radix Ginseng tissue cultured seedling, the segment for cutting into about 5mm × 5mm × 3mm with sterilizing knife, proceed to containing 1/2MS (- N)+
On the solid inducing culture of 2mg/L IBA (in solid inducing culture contain 2% sucrose, 7g/L agar, pH value be 5.8), in
24 ± 2 DEG C, the generation of inducing culture adventitious root under dark condition.Each culture dish is inoculated with 30 explants, inducing culture process
In per three weeks subcultures once, count the indefinite radical that root induction rate and each explant are produced, test is repeated 3 times, makes even
Average, is as a result listed in table 1.
As a result show, after culture in three weeks, all of can induce adventitious root, and per bar root on only induce
1 adventitious root, but through six weeks culture after (situation of inducing culture adventitious root is shown in Figure 1A), averagely per bar root on produce indefinite
Radical is 11.33 (being shown in Table 1).
Embodiment 2
The present embodiment is used for the enrichment culture of the adventitious root for illustrating that Radix Ginseng tissue cultured seedling root induction is produced.
The adventitious root that the Radix Ginseng tissue cultured seedling root induction that inducing culture in embodiment 1 was obtained after six weeks is produced takes out, and is cut into
Fritter is accessed the fluid medium containing 100ml 1/2MS (- N)+2mg/L IBA+20g/L sucrose by the fritter of 1cm sizes
In culture bottle, be placed in shaking table and (60rpm) shaken at a slow speed in 24 ± 2 DEG C of dark conditions, carry out the enrichment culture of adventitious root.Per three
Zhou Jidai once, and counts the fresh weight of adventitious root, and test is repeated 3 times, averages.The adventitious root that culture is obtained takes out, and goes out
The culture fluid of bacterium filter paper adsorption surface, is placed in balance and is weighed, as fresh weight.The rate of increase of adventitious root is calculated, is as a result shown
(see Fig. 2A), after cultivating three weeks in the 1/2MS (- N) culture fluid containing 2mg/LIBA and 20g/L sucrose, the rate of increase is
17.73%；After cultivating six weeks, the rate of increase is significantly improved, and is 27.45%；When, after culture in nine weeks, the rate of increase is declined slightly,
For 25.88%.
Embodiment 3
The present embodiment is used for the enrichment culture of the adventitious root for illustrating that Radix Ginseng tissue cultured seedling root induction is produced.
Enrichment culture is carried out to the adventitious root that Radix Ginseng tissue cultured seedling root induction is produced according to method same as Example 2.Area
It is not only that, the liquid proliferated culture medium for being used is the propagation of the 1/2MS (- N) liquid containing 5mg/L IBA and 20g/L sucrose
Culture medium.The rate of increase of adventitious root is calculated, as a result shows (Fig. 2A)：After enrichment culture three weeks, the rate of increase is 24.4%；Propagation training
After supporting six weeks, the rate of increase is 32.27%, when, after culture in nine weeks, the rate of increase is 22.25%.
Embodiment 4
The present embodiment is used for method of the explanation using Radix Ginseng tissue cultured seedling stem inducing adventitious root.
Take the stem section of Radix Ginseng tissue cultured seedling, the segment for cutting into about 5mm × 5mm × 3mm with sterilizing knife, proceed to containing 1/2MS (-
N) (contain 2% sucrose in solid inducing culture, 7g/L agar, pH value are 5.8), in 24 on the solid medium of+2mg/L IBA
± 2 DEG C, under dark condition, cultivate the generation of inducing adventitious root.Each culture dish is inoculated with 30 explants, per three weeks subcultures once,
And the indefinite radical that root induction rate and each explant are produced is counted, test is repeated 3 times, averages.As a result show, pass through
After crossing culture in three weeks, the explant physical ability induction for having 27.94% can produce adventitious root, and per bar root on only induce 1 indefinite
Root, but after culture in six weeks, all of explant all generates adventitious root (situation of inducing culture adventitious root is shown in Figure 1B), puts down
The indefinite radical produced on per bar root is 10.33 (being shown in Table 1).
Embodiment 5
The present embodiment is used for the enrichment culture method of the adventitious root for illustrating that the induction of Radix Ginseng tissue cultured seedling stem is produced.
The adventitious root that the Radix Ginseng stem induction obtained after six weeks being cultivated in embodiment 4 is produced takes out, and is cut into 1cm sizes, connects
Enter in the culture bottle of the liquid proliferated culture medium containing 100ml 1/2MS (- N)+2mg/L IBA+20g/L sucrose, be placed in shaking table,
24 ± 2 DEG C, dark condition shakes at a slow speed (60rpm), carries out the enrichment culture of adventitious root.Per three weeks subcultures once, and count not
Determine the fresh weight of root, test is repeated 3 times, and averages.The adventitious root that culture is obtained takes out, the culture of sterilizing filter paper adsorption surface
Liquid, is placed in balance and is weighed, as fresh weight.The rate of increase of adventitious root is calculated, is as a result shown (see Fig. 2 B), is being contained 2mg/L
After three weeks are cultivated in the 1/2MS (- N) culture fluid of IBA and 20g/L sucrose, the rate of increase is 270%；After cultivating six weeks, the rate of increase is
188%；When, after culture in nine weeks, the rate of increase is 142%.
Embodiment 6
The present embodiment is used for the enrichment culture of the adventitious root for illustrating that the induction of Radix Ginseng tissue cultured seedling stem is produced.
The adventitious root for producing is induced to carry out enrichment culture on Radix Ginseng tissue cultured seedling stem according to method same as Example 5.Area
It is not only that, the culture medium for being used is 1/2MS (- N) the liquid proliferated culture medium containing 5mg/L IBA and 20g/L sucrose.Meter
The rate of increase of adventitious root is calculated, is as a result shown (see Fig. 2 B), after three weeks, the rate of increase is 295% to enrichment culture；After cultivating six weeks, fresh
The weight rate of increase is 169%, when, after culture in nine weeks, the rate of increase is 126%.
Embodiment 7
The present embodiment is used for the generation that Radix Ginseng tissue cultured seedling leaf inducing adventitious root is described.
Take the blade of Radix Ginseng tissue cultured seedling, the segment for cutting into about 5mm × 5mm × 3mm with sterilizing knife, proceed to containing 1/2MS (-
N) (contain 2% sucrose in solid inducing culture, 7g/L agar, pH value are 5.8), in 24 on the solid medium of+2mg/L IBA
± 2 DEG C, under dark condition, cultivate the generation of inducing adventitious root.Each culture dish is inoculated with 30 explants, per three weeks subcultures once,
And the indefinite radical that root induction rate and each explant are produced is counted, test is repeated 3 times, averages.As a result show, pass through
After crossing culture in three weeks, generation of all of blade all without adventitious root, but (the feelings of inducing culture adventitious root after culture in six weeks
Condition is shown in Fig. 1 C), have 87.78% explant induction generate adventitious root, averagely per bar root on produce indefinite radical be 6.33
Bar (is shown in Table 1).
Embodiment 8
The present embodiment is used for the enrichment culture of the adventitious root for illustrating that the induction of Radix Ginseng tissue culture seedling leaf is produced.
The adventitious root that the leaves of panax ginseng induction obtained after six weeks being cultivated in embodiment 7 is produced takes out, and accesses and contains 100ml
In the culture bottle of the liquid proliferated culture medium of 1/2MS (- N)+2mg/L IBA+20g/L sucrose, shaking table is placed in, 24 ± 2 DEG C, dark
Condition shakes at a slow speed (60rpm), carries out the enrichment culture of adventitious root.Per three weeks subcultures once, and the fresh weight of adventitious root is counted, examination
Test and be repeated 3 times, average.The adventitious root that culture is obtained takes out, and the culture fluid of sterilizing filter paper adsorption surface is placed in balance and enters
Row is weighed, as fresh weight.The rate of increase of adventitious root is calculated, is as a result shown (see Fig. 2 C), containing 2mg/L IBA and 20g/L sugarcanes
In 1/2MS (- N) the liquid proliferated culture medium of sugar, after three weeks, the rate of increase is 713% to enrichment culture；After cultivating six weeks, the rate of increase is
331%；When, after culture in nine weeks, the rate of increase is 77.42%.
Embodiment 9
The present embodiment is used for the enrichment culture of the adventitious root for illustrating that the induction of Radix Ginseng tissue cultured seedling leaf is produced.
The adventitious root for producing is induced to carry out enrichment culture on Radix Ginseng tissue cultured seedling leaf according to method same as Example 8.Area
It is not only that, the culture medium for being used is the 1/2MS (- N) culture fluid containing 5mg/L IBA and 20g/L sucrose, calculates adventitious root
The rate of increase, as a result show (see Fig. 2 C), after three weeks, the rate of increase is 638% to enrichment culture；After cultivating six weeks, the rate of increase is
216%, when, after culture in nine weeks, the rate of increase drops to 83.44%.
Table 1
In table 1, the indefinite radical variation analyses of root induction rate and averagely every piece calluss are checked using T.Different
Letter represents significant difference.During inducing culture 6 weeks, compared with leaf, root and the averagely indefinite radical significant difference of stem, p<0.05.Its
He is p<0.01.
In fig. 2, different letters represent significant difference.With 3 phase of enrichment culture in 1/2MS (- N)+2mg/L IBA
Than, Radix Ginseng tissue cultured seedling root induction adventitious root after 9 weeks enrichment cultures rate of increase significant difference, p<0.05.With 1/2MS (-
N) in+5mg/L IBA, enrichment culture is compared for 6 weeks, the adventitious root that panax ginseng stem segment is induced rate of increase difference after 9 weeks enrichment cultures
Significantly, p<0.05.During the adventitious root enrichment culture 6 weeks of leaves of panax ginseng induction, and in 1/2MS (- N)+2mg/L IBA culture liquid phases
Than, inductivity significant difference in 1/2MS (- N)+5mg/L IBA culture fluid, p<0.05.Other p<0.01.
Comparative example 1
This comparative example is used for the induction for contrasting Radix Ginseng root-derived callus adventitious root.
The root of 5 years raw Radix Ginsengs is taken, is cleaned through clear water, after oven for drying and 75% ethanol surface sterilization 1 minute, super
Sterile scalpel being used in net workbench in the otch of standardized 1-2cm of root surface, and being cut from both sides, interior tissue is cut into
The fritter of 5mm × 5mm × 5mm is inoculated into MS+1mg/L 2, in 4- dichlorphenoxyacetic acid culture medium, carries out the induction of calluss
Culture.Condition of culture is 24 ± 2 DEG C and lucifuge.30g/L sucrose and 7g/L agar are added in culture medium, and pH value is 5.8.Per three
Zhou Jidai is once.The good calluss of growth conditions can be obtained after culture in six weeks.
The calluss of above-mentioned Radix Ginseng root induction are seeded on the solid medium containing 1/2MS (- N)+2mg/LIBA
(contain 2% sucrose in solid inducing culture, concentration is the agar of 0.7 mass %, pH value is the dark bar 5.8), in 24 ± 2 DEG C
The generation of inducing adventitious root is cultivated under part.Each culture dish is inoculated with 30 explants, and counts adventitious root per three weeks subcultures once
The indefinite radical that inductivity and each explant are produced, test are repeated 3 times, average.As a result show, cultivated through three weeks
Afterwards, the Radix Ginseng root-derived callus for having 30% generate adventitious root, averagely per bar root on produce indefinite radical be 1.67, pass through
Six weeks culture after, all of explant is all induced and generates adventitious root, averagely per bar root on produce indefinite radical be 3.67
(being shown in Table 1).
Comparative example 2
This comparative example is used for the propagation for contrasting Radix Ginseng root-derived callus adventitious root.
The adventitious root that the Radix Ginseng callus induction obtained after six weeks being cultivated in comparative example 1 is produced takes out, and access contains
Have in the culture bottle of fluid medium of 100ml 1/2MS (- N)+2mg/L IBA+20g/L sucrose, be placed in shaking table, 24 ± 2 DEG C,
Dark condition shakes at a slow speed (60rpm), carries out the enrichment culture of adventitious root.Per three weeks subcultures once, and the fresh of adventitious root is counted
Weight, test are repeated 3 times, average.The adventitious root that culture is obtained takes out, and the culture fluid of sterilizing filter paper adsorption surface is placed in
Balance is weighed, as fresh weight.As a result show (see Fig. 3), train in the 1/2MS (- N) containing 2mg/L IBA and 20g/L sucrose
After cultivating three weeks in nutrient solution, the rate of increase only about 1%；After cultivating six weeks, the rate of increase has declined, and negative propagation occurs.
Comparative example 3
The present embodiment is used for explanation carries out the propagation of adventitious root using Radix Ginseng root-derived callus.
Enrichment culture is carried out to the adventitious root that Radix Ginseng callus induction is produced according to 2 identical method of comparative example.
Differ only in, the culture medium for being used is the 1/2MS (- N) culture fluid containing 5mg/L IBA and 20g/L sucrose, fresh weight is bred
Multiple result shows (see Fig. 3) that after three weeks, the rate of increase is 3% to enrichment culture；After cultivating six weeks, the rate of increase 105%；Work as process
After culture in nine weeks, the rate of increase is 386.5%.
Although, above the present invention is described in detail with a general description of the specific embodiments,
On the basis of the present invention, it can be made some modifications or improvements, this will be apparent to those skilled in the art.Cause
This, these modifications or improvements, belong to the scope of protection of present invention without departing from theon the basis of the spirit of the present invention.
Claims (5)
1. a kind of ginseng adventitious root proliferative induction method, it is characterised in that comprise the following steps：
1) Radix Ginseng tissue cultured seedling is cut into tissue fritter, using solid inducing culture in the dark that cultivation temperature is 23-25 DEG C
Under the conditions of inducing culture 3-6 week obtain adventitious root；
2) by step 1) adventitious root that obtains cuts into adventitious root fritter, is 23- using liquid proliferated culture medium in cultivation temperature
With the speed concussion and cultivate 3-9 week of 50-80rpm under 25 DEG C of dark condition, the enrichment culture of adventitious root is carried out；
Wherein, the solid inducing culture and liquid proliferated culture medium are to be contained with 1/2MS (- N) culture medium as minimal medium
There are the indolebutyric acid that concentration is 1-10mg/L and the sucrose that concentration is 1.8-2.2 mass %；
Wherein, 1/2MS (- N) culture medium halves the constant MS (- N) culture medium of other compositions, wherein, MS (- N) for a great number of elements
Culture medium is free from the MS culture medium of ammonium nitrate；
Wherein, the Radix Ginseng tissue cultured seedling is through seed sprouting or the tissue cultured seedling of explant isolated culture acquisition, the day of tissue cultured seedling
Age is 28-32 days.
2. root induction enrichment procedure according to claim 1, it is characterised in that the solid inducing culture and liquid
Body proliferated culture medium is to contain indolebutyric acid of the concentration as 2-5mg/L and dense with 1/2MS (- N) culture medium as minimal medium
Spend the sucrose for 1.8-2.2 mass %.
3. root induction enrichment procedure according to claim 1 and 2, it is characterised in that the tissue fritter is Radix Ginseng
Tissue cultured seedling root fritter, Radix Ginseng tissue cultured seedling stem fritter or Radix Ginseng tissue cultured seedling leaf fritter.
4. root induction enrichment procedure according to claim 1 and 2, it is characterised in that the product of the Radix Ginseng tissue cultured seedling
Plant is that Radix Ginseng or Da-maya are joined.
5. root induction enrichment procedure according to claim 3, it is characterised in that the kind of the Radix Ginseng tissue cultured seedling is
Radix Ginseng or Da-maya ginseng.
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| CN105638470A (en) * | 2016-01-09 | 2016-06-08 | 佛山市金蓝领教育科技有限公司 | Ginseng seed germination culture medium |
| CN107343443A (en) * | 2017-08-14 | 2017-11-14 | 高健强 | A kind of Talinum crassifolium large-scale method for producing |
| CN108739377A (en) * | 2018-04-28 | 2018-11-06 | 刘汉石 | A kind of ginseng adventitious root inducing culture and its application |
| CN109729974A (en) * | 2019-02-12 | 2019-05-10 | 天津大学 | A kind of method for tissue culture of Low- temperature culture ginseng adventitious root |
| CN111937748B (en) * | 2020-08-24 | 2022-01-11 | 延边大学 | Culture method for improving content of ginsenoside Rg1 and Re in ginseng adventitious roots |
| CN113647326B (en) * | 2021-01-18 | 2022-11-29 | 山东安然纳米实业发展有限公司 | Method for culturing adventitious roots of ginseng |
| CN113647324B (en) * | 2021-01-18 | 2022-11-29 | 山东安然纳米实业发展有限公司 | Liquid culture medium for culturing ginseng adventitious roots and culture method |
| CN113647327B (en) * | 2021-01-18 | 2022-06-17 | 山东安然纳米实业发展有限公司 | Method for converting ginsenoside in adventitious roots of ginseng |
| CN113647325B (en) * | 2021-01-18 | 2022-11-29 | 山东安然纳米实业发展有限公司 | Liquid culture medium for culturing ginseng adventitious roots and culture method |
| CN112813018B (en) * | 2021-04-02 | 2022-04-12 | 山东安然纳米实业发展有限公司 | Culture method of ginseng stem cells |
| CN113025552B (en) * | 2021-04-02 | 2021-10-26 | 山东安然纳米实业发展有限公司 | Isolated culture method of ginseng stem cells |
| CN113151145B (en) * | 2021-04-20 | 2023-01-24 | 山东安然纳米实业发展有限公司 | Ginseng stem cell separation culture method using biological reaction device |
| CN113025554B (en) * | 2021-04-20 | 2022-06-17 | 山东安然纳米实业发展有限公司 | Method for culturing ginseng stem cells by using biological reaction device |
| CN113151146B (en) * | 2021-04-20 | 2022-07-08 | 山东安然纳米实业发展有限公司 | Ginseng stem cell separation culture method using biological reaction device |
| CN113174360B (en) * | 2021-04-20 | 2023-02-10 | 山东安然纳米实业发展有限公司 | Isolated culture method of ginseng stem cells |
| CN113046294B (en) * | 2021-04-20 | 2022-06-24 | 山东安然纳米实业发展有限公司 | Method for culturing ginseng stem cells |
| CN113025553B (en) * | 2021-04-20 | 2022-06-14 | 山东安然纳米实业发展有限公司 | Method for culturing ginseng stem cells by using biological reaction device |
| CN113151144B (en) * | 2021-04-20 | 2022-05-31 | 山东安然纳米实业发展有限公司 | Isolated culture method of ginseng stem cells |
| CN113046296B (en) * | 2021-04-20 | 2023-02-10 | 山东安然纳米实业发展有限公司 | Method for culturing ginseng stem cells by using biological reaction device |
| CN114403005A (en) * | 2022-01-27 | 2022-04-29 | 上海瑞帝安生物科技有限公司 | Ginseng adventitious root induction method and application of extract in cosmetics |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1285138A (en) * | 1999-08-23 | 2001-02-28 | 赵寿经 | Method for producing ginsent hairy root and ginsent seponin by hairy root agricillin A4 bacterial strain inducing explant of ginsent root |
| KR20040027818A (en) * | 2004-03-10 | 2004-04-01 | (주)바이오피아 | Mass Production Method of the Ginseng Saponin by the Treatment with Mevalonic Acid |
| CN101473788A (en) * | 2008-12-31 | 2009-07-08 | 上海交通大学 | Method for inducing and cultivating adventitious root of tetraploid Radix pseudostellariae |
| CN101606487A (en) * | 2009-07-13 | 2009-12-23 | 天津大学 | Tissue culture method of ginseng adventitious root |
| CN103141394A (en) * | 2013-03-28 | 2013-06-12 | 延边大学 | Method for culturing adventitious roots of oplopanax elatus Nakai by utilizing bioreactor |
| CN104026019A (en) * | 2013-02-04 | 2014-09-10 | 南京工业大学大丰海洋产业研究院 | Method for continuously obtaining secondary ginseng hairy root metabolites through tissue culture |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP3737417B2 (en) * | 2001-01-19 | 2006-01-18 | パク、キ−ヨエウ | Method for mass growth of adventitious roots of ginseng, camphor ginseng, and ginseng by tissue culture and improvement of saponin content |
-
2014
- 2014-11-26 CN CN201410698528.0A patent/CN104472359B/en not_active Expired - Fee Related
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1285138A (en) * | 1999-08-23 | 2001-02-28 | 赵寿经 | Method for producing ginsent hairy root and ginsent seponin by hairy root agricillin A4 bacterial strain inducing explant of ginsent root |
| KR20040027818A (en) * | 2004-03-10 | 2004-04-01 | (주)바이오피아 | Mass Production Method of the Ginseng Saponin by the Treatment with Mevalonic Acid |
| CN101473788A (en) * | 2008-12-31 | 2009-07-08 | 上海交通大学 | Method for inducing and cultivating adventitious root of tetraploid Radix pseudostellariae |
| CN101606487A (en) * | 2009-07-13 | 2009-12-23 | 天津大学 | Tissue culture method of ginseng adventitious root |
| CN104026019A (en) * | 2013-02-04 | 2014-09-10 | 南京工业大学大丰海洋产业研究院 | Method for continuously obtaining secondary ginseng hairy root metabolites through tissue culture |
| CN103141394A (en) * | 2013-03-28 | 2013-06-12 | 延边大学 | Method for culturing adventitious roots of oplopanax elatus Nakai by utilizing bioreactor |
Non-Patent Citations (1)
| Title |
|---|
| 药用植物人参的组织培养研究进展;左北梅等;《中国现代中药》;20120131;第14卷(第1期);第34-37页 * |
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