CN105191790A - In-vitro culturing method for rhodiola dumulosa - Google Patents
In-vitro culturing method for rhodiola dumulosa Download PDFInfo
- Publication number
- CN105191790A CN105191790A CN201510514981.6A CN201510514981A CN105191790A CN 105191790 A CN105191790 A CN 105191790A CN 201510514981 A CN201510514981 A CN 201510514981A CN 105191790 A CN105191790 A CN 105191790A
- Authority
- CN
- China
- Prior art keywords
- medium
- final concentration
- adventitious bud
- naa
- solid culture
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Landscapes
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses an in-vitro culturing method for rhodiola dumulosa. The in-vitro culturing method for the rhodiola dumulosa comprises the following steps that 1, stem segments of the rhodiola dumulosa are inoculated onto a callus tissue inducing medium to be cultured to obtain callus tissue; 2, the callus tissue obtained in the step 1 is inoculated onto an adventitious bud inducing medium to be cultured to obtain adventitious buds; 3, the adventitious buds obtained in the step 2 are inoculated onto an adventitious bud multiplication medium to be cultured to obtain multiple buds; 4, the multiple buds obtained in the step 3 are inoculated onto a rooting medium to be cultured to obtain regeneration seedlings. In-vitro culturing of the rhodiola dumulosa is performed by adopting the method, the rooting rate can reach 100 percent, the average rooting number is about 10-15, and the root length is 2-3 cm; regeneration plants are transplanted after rooting is achieved, and the survival rate reaches 90 percent. According to the in-vitro culturing method for the rhodiola dumulosa, an in-vitro culturing regeneration system of the rhodiola dumulosa is built, and the important significance is achieved for effectively protecting and utilizing wild rhodiola dumulosa resources.
Description
Technical field
The invention belongs to field of plant tissue culture, relate to a kind of in-vitro culture method of Hericiumerinaceus Pers.
Background technology
Hericiumerinaceus Pers Rhodioladumulosa (Franch.) S.H.Fu, has another name called Radix Rhodiolae Dumulosae, is Crassulaceae herbaceos perennial.Originate in Northwest Sichuan, Qinghai, Gansu, Shaanxi, Hubei, Shanxi, Hebei, the Inner Mongol, Jilin, be born on the hillside stone of height above sea level 1600-3900 rice.
Hericiumerinaceus Pers is Tibetan's rare medicinal herbs, have another name called " sweeping Luo Maerbu ", modern medicine confirms: Hericiumerinaceus Pers has " adaptation former state " effect that the similar traditional Chinese medical science " is strengthened the body resistance to consolidate the constitution ", namely impels the special regulatory function that the ill-conditioned index caused by different pathogeny changes to normal condition.It nourishes, keep fit, diseases prevention and anti-aging effects are rare in known tonic.The effect of the enhancing immunologic function of Hericiumerinaceus Pers is better than ginseng, is the pure green medicinal plant also more precious than ginseng.Collar is medicinal, has the effect of kidney tonifying, mental-tranquilization, regulating menstruation and activating blood, improving eyesight.
There is not been reported for the current tissue cultures about Hericiumerinaceus Pers.
Summary of the invention
An object of the present invention is to provide a kind of in-vitro culture method of Hericiumerinaceus Pers.
The in-vitro culture method of Hericiumerinaceus Pers provided by the present invention, specifically can comprise the steps:
(1) the stem section of Hericiumerinaceus Pers is inoculated on callus inducing medium, carries out induction of callus, obtain callus;
(2) step (1) gained callus is inoculated on adventitious bud induction culture base, carries out adventitious bud induction culture, obtain indefinite bud;
(3) step (2) gained indefinite bud is inoculated on adventitious bud proliferation medium, carries out adventitious bud proliferation cultivation, obtain Multiple Buds;
(4) step (3) gained Multiple Buds is inoculated on root media, carries out culture of rootage, obtain regrowth.
Described callus inducing medium is the medium obtained add 6-BA, NAA and 2,4-D in MS solid culture medium after; The final concentration of 6-BA to be the final concentration of 1.0-1.5mg/L, NAA be 0.2-0.5mg/L in described callus inducing medium, 2,4-D final concentration be 0.2-0.5mg/L; PH5.8.
Described adventitious bud induction culture base is medium shown in following (a1) or (a2):
(a1) medium obtained after adding 6-BA and NAA in MS solid culture medium; In described adventitious bud induction culture base, the final concentration of 6-BA is the final concentration of 0.5-1.0mg/L, NAA is 0.1-0.5mg/L; PH5.8;
(a2) medium obtained after adding TDZ and NAA in MS solid culture medium; In described adventitious bud induction culture base, the final concentration of TDZ is the final concentration of 0.5-1.0mg/L, NAA is 0.5-1.0mg/L; PH5.8.
Described adventitious bud proliferation medium is the medium obtained add TDZ in MS solid culture medium after; In described adventitious bud proliferation medium, the final concentration of TDZ is 0.5-2.0mg/L; PH5.8.
Described root media is the medium obtained add IBA in MS solid culture medium after; In described root media, the final concentration of IBA is 1.0-2.0mg/L; PH5.8.
Further, the final concentration that in described callus inducing medium, the final concentration of 6-BA is specially 0.85mg/L, NAA be specially 0.34mg/L, 2,4-D final concentration be specially 0.33mg/L; In described adventitious bud proliferation medium, the final concentration of TDZ is specially 0.5-1.0mg/L, as 0.5mg/L; In described root media, the final concentration of IBA is specially 1.0mg/L.
Wherein, the carbon source in described MS solid culture medium is sucrose; Gel in described MS solid culture medium is agar.The final concentration of described sucrose in described MS solid culture medium is specially 30g/L; The final concentration of described agar in described MS solid culture medium is specially 7g/L.
In the process, carry out carrying out in the condition of described induction of callus, step (2) condition of carrying out described adventitious bud proliferation cultivation in the condition of described adventitious bud induction culture, step (3) in step (1), and in step (4), carry out the condition of described culture of rootage, all can be: temperature 25 ± 2 DEG C, light application time 12h/ days, intensity of illumination 30 ~ 40mmolm
-2s
-1.
In addition, carry out carrying out in the cycle of described induction of callus, step (2) cycle of carrying out described adventitious bud proliferation cultivation in the cycle of described adventitious bud induction culture, step (3) in step (1), and the cycle of carrying out described culture of rootage in step (4) is 35 days.
In the step (1) of said method, the stem section of described Hericiumerinaceus Pers is the stem section after sterilization; Described sterilization specifically can be: by the stem section running water 20 minutes of Hericiumerinaceus Pers to be sterilized, by the alcohol disinfecting 10s that flushed stem section volume fraction is 75%, with aseptic water washing (as rinsed 3 times), being the mercuric chloride aqueous solution sterilization 2min of 0.1% again with mass fraction, finally using aseptic water washing (as rinsed 3-5 time).
In the above-mentioned methods, the step of described regrowth being carried out hardening and transplanting can also be comprised.Be specially: first by described regrowth in desinfection chamber hardening after 1 week, clean the medium of root, then transplant to peat soil: perlite: in the mixed-matrix of sand=4:3:2 (volume ratio), be placed in the environment that relative air humidity is 80% – 90%, temperature is 20 DEG C, water every day 2 times.
Another object of the present invention is to provide a kind of medium of the cultured in vitro for Hericiumerinaceus Pers.
The medium of the cultured in vitro for Hericiumerinaceus Pers provided by the present invention, is made up of all or part of in following medium:
(I) callus inducing medium:
Described callus inducing medium is the medium obtained add 6-BA, NAA and 2,4-D in MS solid culture medium after; The final concentration of 6-BA to be the final concentration of 1.0-1.5mg/L, NAA be 0.2-0.5mg/L in described callus inducing medium, 2,4-D final concentration be 0.2-0.5mg/L; PH5.8;
(II) adventitious bud induction culture base:
Described adventitious bud induction culture base is medium shown in following (a1) or (a2):
(a1) medium obtained after adding 6-BA and NAA in MS solid culture medium; In described adventitious bud induction culture base, the final concentration of 6-BA is the final concentration of 0.5-1.0mg/L, NAA is 0.1-0.5mg/L; PH5.8;
(a2) medium obtained after adding TDZ and NAA in MS solid culture medium; In described adventitious bud induction culture base, the final concentration of TDZ is the final concentration of 0.5-1.0mg/L, NAA is 0.5-1.0mg/L; PH5.8;
(III) adventitious bud proliferation medium:
Described adventitious bud proliferation medium is the medium obtained add TDZ in MS solid culture medium after; In described adventitious bud proliferation medium, the final concentration of TDZ is 0.5-2.0mg/L; PH5.8;
(IV) root media:
Described root media is the medium obtained add IBA in MS solid culture medium after; In described root media, the final concentration of IBA is 1.0-2.0mg/L; PH5.8.
Further, the final concentration that in described callus inducing medium, the final concentration of 6-BA is specially 0.85mg/L, NAA be specially 0.34mg/L, 2,4-D final concentration be specially 0.33mg/L; In described adventitious bud proliferation medium, the final concentration of TDZ is specially 0.5-1.0mg/L, as 0.5mg/L; In described root media, the final concentration of IBA is specially 1.0mg/L.
Wherein, the carbon source in described MS solid culture medium is sucrose; Gel in described MS solid culture medium is agar.
The final concentration of described sucrose in described MS solid culture medium is 30g/L; The final concentration of described agar in described MS solid culture medium is 7g/L.
In the in-vitro culture method of above Hericiumerinaceus Pers provided by the present invention, and in the medium for Hericiumerinaceus Pers cultured in vitro provided by the present invention, the pH5.8 of described MS solid culture medium, solvent is water, solute and concentration as follows:
A. a large amount of: NH
4nO
31.65g/L, KNO
31.9g/L, MgSO
4 .7H
2o0.37g/L, KH
2pO
40.17g/L, CaCl
22H
2o0.44g/L;
B. trace: KI0.83mg/L, H
3bO
36.2mg/L, MnSO
4 .4H
2o22.3mg/L, ZnSO
4 .7H
2o8.6mg/L, Na
2moO
4 .2H
2o0.25mg/L, CuSO
4 .5H
2o0.025mg/L, CoCl
2 .6H
2o0.025mg/L;
C. vitamin: nicotinic acid 0.5mg/L, pyridoxine hydrochloride 0.5mg/L, thiamine hydrochloride 0.1mg/L, glycine 2mg/L;
D. molysite: FeSO
4 .7H
2o27.8mg/L, Na
2eDTA37.3mg/L;
E. inositol 0.1g/L;
F. sucrose 30g/L;
G. agar 7g/L.
Each concentration is the final concentration of respective components in described MS solid culture medium above.
Described medium is using stem section as explant, the application carried out in Hericiumerinaceus Pers cultured in vitro also belongs to protection scope of the present invention.
In the present invention, the length of described stem section can be 0.5cm.
The present invention carries out tissue cultures and plant regeneration research using the stem section of Hericiumerinaceus Pers as explant, establish the in-vitro propagate system of Hericiumerinaceus Pers.Result of study shows, the optimal medium of Hericiumerinaceus Pers callus induction is MS+6-BA0.85mg/L+NAA0.34mg/L+2,4-D0.33mg/L, and inductivity can reach 90%.The inducing culture of indefinite bud is MS+6-BA0.5-1.0mg/L+NAA0.1-0.5mg/L or MS+TDZ0.5-1.0mg/L+NAA0.5-1.0mg/L, but the inductivity of indefinite bud is all lower.Transfer on the MS medium containing TDZ0.5-2.0mg/L by the callus of band indefinite bud, indefinite bud can the propagation of rapid, high volume, grows fine.On the MS medium adding IBA1.0-2.0mg/L, adventitious bud rooting rate can reach 100%, number of on average taking root about 10 ~ 15, the long 2 ~ 3cm of root.After about 30d, regeneration plant starts to bloom.After taking root, regeneration plant is transplanted, survival rate 90%.The present invention is for available protecting and utilize wild Hericiumerinaceus Pers resource significant, is beneficial to the large-scale production promoting Hericiumerinaceus Pers simultaneously.
Embodiment
The experimental technique used in following embodiment if no special instructions, is conventional method.
Material used in following embodiment, reagent etc., if no special instructions, all can obtain from commercial channels.
The pH of MS solid culture medium involved in following embodiment is 5.8, and concrete formula is as shown in table 1:
Table 1MS solid culture medium composition (pH5.8)
| Compound title | Concentration (mg/L) in medium |
| CaCl 2·2H 2O | 440 |
| KH 2PO 4 | 170 |
| MgSO 4·7H 2O | 370 |
| NH 4NO 3 | 1650 |
| KNO 3 | 1900 |
| KI | 0.83 |
| CoCl 2·6H 2O | 0.025 |
| H 3BO 3 | 6.2 |
| Na 2MoO 4·2H 2O | 0.25 |
| MnSO 4·4H 2O | 22.3 |
| CuSO 4·5H 2O | 0.025 |
| ZnSO 4·7H 2O | 8.6 |
| Na 2EDTA | 37.3 |
| FeSO 4·7H 2O | 27.8 |
| Thiamine hydrochloride | 0.1 |
| Pyridoxine hydrochloride | 0.5 |
| Nicotinic acid | 0.5 |
| Inositol | 100 |
| Glycine | 2.0 |
| Sucrose | 30000 |
| Agar | 7000 |
The cultured in vitro of embodiment 1, Hericiumerinaceus Pers
One, explant selection with disinfect
Hericiumerinaceus Pers [Rhodioladumulosa (Franch.) S.H.Fu] picks up from the Dongling Mountain of height above sea level about 2000m in July, 2012, and confirm according to " Chinese Plants will " described Hericiumerinaceus Pers morphological feature standard, material object conforms to title.Choose the stem section that Hericiumerinaceus Pers children is tender, use tap water 20mins, superclean bench is first used the alcohol surface sterilization 10s of 75% (volume fraction), with aseptic water washing 3 times, 0.1% (mass fraction) mercuric chloride is used to sterilize 2mins again, with aseptic water washing 3 ~ 5 times.
Two, the Fiber differentiation of callus
The aseptic stem section of disinfecting through step one is cut into the segment that length is 0.5cm respectively, is inoculated on callus inducing medium (MS+6-BA1.0mg/L+NAA0.2mg/L+2,4-D0.1mg/L), carries out induction of callus.Wherein, callus inducing medium (MS+6-BA1.0mg/L+NAA0.2mg/L+2,4-D0.1mg/L) for add 6-BA, NAA and 2 in MS solid culture medium, the medium obtained after 4-D; The final concentration of 6-BA to be the final concentration of 1.0mg/L, NAA be 0.2mg/L in described callus inducing medium, 2,4-D final concentration be 0.1mg/L; PH5.8.Condition of culture is temperature 25 ± 2 DEG C, light application time 12h/ days, intensity of illumination 30 ~ 40mmolm
-2s
-1; Cultivation cycle is 35 days.
Result shows: callus inducing medium (MS+6-BA1.0mg/L+NAA0.2mg/L+2,4-D0.1mg/L) can induce callus, but out of order, produce callus amount less, callus induction rate is only 63%.
The present inventor is further at callus inducing medium (MS+6-BA1.0mg/L+NAA0.2mg/L+2, the basis of 4-D0.1mg/L) filling a prescription use response surface optimization method seek the best concentration ratio of 6-BA, NAA and this three plant growth regulators of 2,4-D adjustment evoked callus.
By analyzing the condition obtaining the optimization of Hericiumerinaceus Pers callus induction be: 6-BA0.85mg/L, NAA0.34mg/L, 2,4-D0.33mg/L, its theoretical callus induction rate is 91.61%, 3 demonstration tests (condition of culture is the same) are carried out according to optimal conditions, actual average callus induction rate is 90.03%, substantially conforms to theoretical value.The optimal medium of Hericiumerinaceus Pers callus induction is MS+6-BA0.85mg/L+NAA0.34mg/L+2,4-D0.33mg/L, is the medium obtained add 6-BA, NAA and 2,4-D in MS solid culture medium after; The final concentration of 6-BA to be the final concentration of 0.85mg/L, NAA be 0.34mg/L in described callus inducing medium, 2,4-D final concentration be 0.33mg/L; PH5.8.
Three, the Fiber differentiation of indefinite bud
The callus of Hericiumerinaceus Pers that step 2 obtains is inoculated in containing variable concentrations 6-BA (0,0.5,1.0,2.0,4.0mg/L) and NAA (0,0.1,0.5,1.0,1.5mg/L) MS solid culture medium in, or be inoculated in containing variable concentrations TDZ (0,0.5,1.0,2.0,4.0mg/L) and NAA (0,0.1,0.5,1.0,1.5mg/L) MS solid culture medium in cultivate.Condition of culture is temperature 25 ± 2 DEG C, light application time 12h/ days, intensity of illumination 30 ~ 40mmolm
-2s
-1; The adventitious bud induction culture cycle is 35 days.
Result shows: when " 6-BA is 0.5-1.0mg/L, NAA is 0.1-0.5mg/L " or " TDZ is 0.5-1.0mg/L, NAA is 0.5-1.0mg/L ", can induce and produce indefinite bud, but the bud quantity produced is little, and can not continue to grow tall.And all can not produce indefinite bud under other concentration.
Namely adventitious bud induction culture base can be medium shown in (a1) or (a2) as follows:
(a1) medium obtained after adding 6-BA and NAA in MS solid culture medium; In described adventitious bud induction culture base, the final concentration of 6-BA is the final concentration of 0.5-1.0mg/L, NAA is 0.1-0.5mg/L; PH5.8.The inductivity of its indefinite bud is 12.04%-40%.
(a2) medium obtained after adding TDZ and NAA in MS solid culture medium; In described adventitious bud induction culture base, the final concentration of TDZ is the final concentration of 0.5-1.0mg/L, NAA is 0.5-1.0mg/L; PH5.8.The inductivity of its indefinite bud is 10%-20%.
Four, the Multiplying culture of indefinite bud
The MS solid culture medium that indefinite bud step 3 induced together forwards to only containing TDZ together with callus carries out Multiplying culture, and TDZ concentration is arranged as Gradient 0mg/L, 0.5mg/L, 1.0mg/L, 2.0mg/L, 4.0mg/L.Condition of culture is temperature 25 ± 2 DEG C, light application time 12h/ days, intensity of illumination 30 ~ 40mmolm
-2s
-1; Adventitious bud proliferation cultivation cycle is 35 days.
Add up the cultivation effect (amplification rate) of different TDZ concentration cultures to indefinite bud, as shown in table 2.Visible MS+TDZ0.5-2.0mg/L is the appropriate media of adventitious bud proliferation.The medium that adventitious bud proliferation medium (MS+TDZ0.5-2.0mg/L) obtains after being and adding TDZ in MS solid culture medium; In described adventitious bud proliferation medium, the final concentration of TDZ is 0.5-2.0mg/L; PH5.8.
Table 2TDZ is to the multiplication effect of indefinite bud
Note: amplification rate (%)=(forming the block number/inoculation block number of Multiple Buds) × 100% " amplification rate " row, represent significant difference (P<0.05) between different lowercase.
Five, culture of rootage obtains regrowth
Root media (MS+IBA1.0-2.0mg/L): the medium obtained add IBA in MS solid culture medium after; In described root media, the final concentration of IBA is 1.0mg/L; PH5.8.
The Multiple Buds that step 4 obtains is inoculated in root media, carries out culture of rootage.Condition of culture is temperature 25 ± 2 DEG C, light application time 12h/ days, intensity of illumination 30 ~ 40mmolm
-2s
-1; The culture of rootage cycle is 35 days.
Result shows: when cultivating about two weeks, Multiple Buds starts to occur white little hair root, and distribute radially, root hair is many, and rooting rate reaches 100%.Number of on average taking root is 12, and average root is long is 3.15cm.Culture of rootage obtained regrowth after 35 days.
Six, hardening and transplanting
The bottleneck of the triangular flask that step 5 gained regrowth is housed is opened, first by regrowth in desinfection chamber hardening after 1 week, careful taking-up regrowth cleans the medium of root, reduce the damage to root system as far as possible, then transplant to peat soil: perlite: in the mixed-matrix of sand=4:3:2 (volume ratio), be placed in the environment that relative air humidity is 80% – 90%, temperature is 20 DEG C, water every day 2 times, survival rate 90% after 30 days.
Claims (10)
1. an in-vitro culture method for Hericiumerinaceus Pers, comprises the steps:
(1) the stem section of Hericiumerinaceus Pers is inoculated on callus inducing medium, carries out induction of callus, obtain callus;
(2) step (1) gained callus is inoculated on adventitious bud induction culture base, carries out adventitious bud induction culture, obtain indefinite bud;
(3) step (2) gained indefinite bud is inoculated on adventitious bud proliferation medium, carries out adventitious bud proliferation cultivation, obtain Multiple Buds;
(4) step (3) gained Multiple Buds is inoculated on root media, carries out culture of rootage, obtain regrowth.
2. method according to claim 1, is characterized in that: described callus inducing medium is the medium obtained add 6-BA, NAA and 2,4-D in MS solid culture medium after; The final concentration of 6-BA to be the final concentration of 1.0-1.5mg/L, NAA be 0.2-0.5mg/L in described callus inducing medium, 2,4-D final concentration be 0.2-0.5mg/L;
Or
Described adventitious bud induction culture base is medium shown in following (a1) or (a2):
(a1) medium obtained after adding 6-BA and NAA in MS solid culture medium; In described adventitious bud induction culture base, the final concentration of 6-BA is the final concentration of 0.5-1.0mg/L, NAA is 0.1-0.5mg/L;
(a2) medium obtained after adding TDZ and NAA in MS solid culture medium; In described adventitious bud induction culture base, the final concentration of TDZ is the final concentration of 0.5-1.0mg/L, NAA is 0.5-1.0mg/L;
Or
Described adventitious bud proliferation medium is the medium obtained add TDZ in MS solid culture medium after; In described adventitious bud proliferation medium, the final concentration of TDZ is 0.5-2.0mg/L;
Or
Described root media is the medium obtained add IBA in MS solid culture medium after; In described root media, the final concentration of IBA is 1.0-2.0mg/L.
3. method according to claim 2, is characterized in that: the final concentration of 6-BA to be the final concentration of 0.85mg/L, NAA be 0.34mg/L in described callus inducing medium, 2,4-D final concentration be 0.33mg/L; Or
In described adventitious bud proliferation medium, the final concentration of TDZ is 0.5-1.0mg/L; Or
In described root media, the final concentration of IBA is 1.0mg/L.
4., according to described method arbitrary in claim 1-3, it is characterized in that: the carbon source in described MS solid culture medium is sucrose; Gel in described MS solid culture medium is agar;
Concrete, the final concentration of described sucrose in described MS solid culture medium is 30g/L; The final concentration of described agar in described MS solid culture medium is 7g/L.
5. according to described method arbitrary in claim 1-4, it is characterized in that: the condition of carrying out carrying out in the condition of described induction of callus, step (2) carrying out in the condition of described adventitious bud induction culture, step (3) described adventitious bud proliferation cultivation in step (1), and in step (4), carry out the condition of described culture of rootage, be: temperature 25 ± 2 DEG C, light application time 12h/ days, intensity of illumination 30 ~ 40mmolm
-2s
-1.
6., for the medium of the cultured in vitro of Hericiumerinaceus Pers, be made up of all or part of in following medium:
(I) callus inducing medium:
Described callus inducing medium is the medium obtained add 6-BA, NAA and 2,4-D in MS solid culture medium after; The final concentration of 6-BA to be the final concentration of 1.0-1.5mg/L, NAA be 0.2-0.5mg/L in described callus inducing medium, 2,4-D final concentration be 0.2-0.5mg/L;
(II) adventitious bud induction culture base:
Described adventitious bud induction culture base is medium shown in following (a1) or (a2):
(a1) medium obtained after adding 6-BA and NAA in MS solid culture medium; In described adventitious bud induction culture base, the final concentration of 6-BA is the final concentration of 0.5-1.0mg/L, NAA is 0.1-0.5mg/L;
(a2) medium obtained after adding TDZ and NAA in MS solid culture medium; In described adventitious bud induction culture base, the final concentration of TDZ is the final concentration of 0.5-1.0mg/L, NAA is 0.5-1.0mg/L;
(III) adventitious bud proliferation medium:
Described adventitious bud proliferation medium is the medium obtained add TDZ in MS solid culture medium after; In described adventitious bud proliferation medium, the final concentration of TDZ is 0.5-2.0mg/L;
(IV) root media:
Described root media is the medium obtained add IBA in MS solid culture medium after; In described root media, the final concentration of IBA is 1.0-2.0mg/L.
7. medium according to claim 6, is characterized in that: the final concentration of 6-BA to be the final concentration of 0.85mg/L, NAA be 0.34mg/L in described callus inducing medium, 2,4-D final concentration be 0.33mg/L; Or
In described adventitious bud proliferation medium, the final concentration of TDZ is 0.5-1.0mg/L; Or
In described root media, the final concentration of IBA is 1.0mg/L.
8. the medium according to claim 6 or 7, is characterized in that: the carbon source in described MS solid culture medium is sucrose; Gel in described MS solid culture medium is agar;
Concrete, the final concentration of described sucrose in described MS solid culture medium is 30g/L; The final concentration of described agar in described MS solid culture medium is 7g/L.
9. in claim 6-8, arbitrary described medium is carrying out the application in Hericiumerinaceus Pers cultured in vitro using stem section as explant.
10., according to described method arbitrary in claim 1-9 or medium or application, it is characterized in that: the length of described stem section is 0.5cm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510514981.6A CN105191790B (en) | 2015-08-20 | 2015-08-20 | In-vitro culturing method for rhodiola dumulosa |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510514981.6A CN105191790B (en) | 2015-08-20 | 2015-08-20 | In-vitro culturing method for rhodiola dumulosa |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105191790A true CN105191790A (en) | 2015-12-30 |
| CN105191790B CN105191790B (en) | 2017-05-24 |
Family
ID=54940083
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510514981.6A Active CN105191790B (en) | 2015-08-20 | 2015-08-20 | In-vitro culturing method for rhodiola dumulosa |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105191790B (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105850743A (en) * | 2016-05-04 | 2016-08-17 | 沈阳农业大学 | Winged euonymus stem adventitious bud inducing method |
| CN106520665A (en) * | 2016-11-15 | 2017-03-22 | 天津市博爱生物药业有限公司 | Culture medium for rhodiola rosea cell |
| CN107347650A (en) * | 2017-08-28 | 2017-11-17 | 江苏丰收大地种业发展有限公司 | A kind of Radix Rhodiolae tissue culture propagation method |
| CN108040872A (en) * | 2017-11-29 | 2018-05-18 | 云南省农业科学院花卉研究所 | A kind of Vitro Quick Reproduction cultural method of feverfew |
| CN109452174A (en) * | 2018-12-21 | 2019-03-12 | 西藏诺迪康药业股份有限公司 | The fast propagating culture medium and propagation method of rhodiola |
| CN109819895A (en) * | 2019-03-25 | 2019-05-31 | 刘艳涛 | A kind of method for tissue culture of kalanchoe daigremontiana |
| CN112616674A (en) * | 2021-01-08 | 2021-04-09 | 南京农业大学 | Induced proliferation culture medium for tissue culture and rapid propagation of rhodiola rosea and tissue culture and rapid propagation method |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101773070A (en) * | 2010-01-19 | 2010-07-14 | 烟台汇鹏生物科技有限公司 | Rhodiola rosea callus culture method |
| CN101836585A (en) * | 2009-03-19 | 2010-09-22 | 中国科学院过程工程研究所 | Tissue-culture seedling raising method of rhodiola crenulata |
-
2015
- 2015-08-20 CN CN201510514981.6A patent/CN105191790B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101836585A (en) * | 2009-03-19 | 2010-09-22 | 中国科学院过程工程研究所 | Tissue-culture seedling raising method of rhodiola crenulata |
| CN101773070A (en) * | 2010-01-19 | 2010-07-14 | 烟台汇鹏生物科技有限公司 | Rhodiola rosea callus culture method |
Non-Patent Citations (3)
| Title |
|---|
| 张晓艳等: "八宝景天的组织培养与快速繁殖", 《吉林师范大学学报》 * |
| 张瑜等: "高山红景天的组织培养与快速繁殖", 《黑龙江医药》 * |
| 沈梦圆等: "激素对四裂红景天愈伤组织诱导及其继代培养的影响", 《生物加工过程》 * |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105850743A (en) * | 2016-05-04 | 2016-08-17 | 沈阳农业大学 | Winged euonymus stem adventitious bud inducing method |
| CN105850743B (en) * | 2016-05-04 | 2017-10-20 | 沈阳农业大学 | A kind of winged euonymus stem section adventitious bud inducing method |
| CN106520665A (en) * | 2016-11-15 | 2017-03-22 | 天津市博爱生物药业有限公司 | Culture medium for rhodiola rosea cell |
| CN107347650A (en) * | 2017-08-28 | 2017-11-17 | 江苏丰收大地种业发展有限公司 | A kind of Radix Rhodiolae tissue culture propagation method |
| CN108040872A (en) * | 2017-11-29 | 2018-05-18 | 云南省农业科学院花卉研究所 | A kind of Vitro Quick Reproduction cultural method of feverfew |
| CN108040872B (en) * | 2017-11-29 | 2021-03-30 | 云南省农业科学院花卉研究所 | In-vitro rapid propagation culture method for white chrysanthemum |
| CN109452174A (en) * | 2018-12-21 | 2019-03-12 | 西藏诺迪康药业股份有限公司 | The fast propagating culture medium and propagation method of rhodiola |
| CN109819895A (en) * | 2019-03-25 | 2019-05-31 | 刘艳涛 | A kind of method for tissue culture of kalanchoe daigremontiana |
| CN112616674A (en) * | 2021-01-08 | 2021-04-09 | 南京农业大学 | Induced proliferation culture medium for tissue culture and rapid propagation of rhodiola rosea and tissue culture and rapid propagation method |
| CN112616674B (en) * | 2021-01-08 | 2022-04-29 | 南京农业大学 | Induced proliferation culture medium for tissue culture and rapid propagation of rhodiola rosea and tissue culture and rapid propagation method |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105191790B (en) | 2017-05-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105191790B (en) | In-vitro culturing method for rhodiola dumulosa | |
| CN104686345A (en) | Tissue culture rapid propagation method of liquorice | |
| CN104126506B (en) | A kind of method for tissue culture of American Red rocket Lagerstroemia indica L. | |
| CN104686338A (en) | In-vitro culture technique of anther of angelica dahurica | |
| CN104206270A (en) | Tissue culture and rhizome expansion method of Curcuma hybrida 'Red Torch' | |
| CN103270950B (en) | Chrysanthemum simplified tissue culturing method | |
| CN105145365B (en) | A kind of tissue culture propagation method of China Caulis cyatheae spinulosae green spheroids approach | |
| CN103828716B (en) | The method for tissue culture of maiden China pink | |
| CN104429941A (en) | In-vitro rapid propagation technique of melaleuca alternifolia | |
| KR100883188B1 (en) | Induction Method of Protocorm like Body from Phalaenopsis Adult Plant | |
| CN104488723A (en) | Tissue-culture and rapid-propagation method of epimedium koreanum nakai | |
| CN105532463A (en) | Proliferation medium for increasing saffron crocus bud proliferation rate | |
| CN103548695B (en) | A kind of meadowrueleaf corydalis root quick breeding method for tissue culture | |
| CN102630569A (en) | Tissue culture method of Gentiana dahurica Fisch for rapid in vitro reproduction | |
| CN102342248A (en) | Potato stem apex detoxification breeding method and double-layer culture medium | |
| CN110663555B (en) | Rapid propagation method of odorous jasminum grandiflorum | |
| CN104304034A (en) | Induction culture method and special induction culture medium for somatic embryo calluses of picea schrenkiana | |
| CN104686346A (en) | Tissue culture and rapid propagation method for divaricate saposhnikovia root | |
| CN104782494A (en) | Thymus mongolicus Ronn. tissue culture rapid propagation method | |
| CN104904594B (en) | A kind of red palm pollen cultures method | |
| CN101843219B (en) | Use of young scape as explant for quick propagation and transgenosis of Chinese sacred lily | |
| CN108094200A (en) | A kind of be heat-treated combines the breeding method that stem apex stripping acquisition peace ancestral spends detoxic seedling | |
| CN105145364A (en) | Tissue culture propagation method for obtaining cibotium barometz green globular body | |
| CN106605596A (en) | Method for mass propagation of lycoris aurea through somatic embryogenesis | |
| CN103081805B (en) | Method for efficiently culturing tissues and industrially propagating robinia idaho |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |