CN105941150A - Method for germinating and propagating cinnamomum camphora 'Yongjin' clusters - Google Patents
Method for germinating and propagating cinnamomum camphora 'Yongjin' clusters Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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Abstract
The invention discloses a method for germinating and propagating cinnamomum camphora 'Yongjin' clusters. The method includes steps of carrying out sterile treatment on young cinnamomum camphora 'Yongjin' sprouts with auxiliary buds yet to be germinated, and then inoculating the young cinnamomum camphora 'Yongjin' sprouts in induction culture media to obtain cluster buds; transferring the young cinnamomum camphora 'Yongjin' sprouts with the cluster buds into propagation culture media, cultivating the young cinnamomum camphora 'Yongjin' sprouts in the propagation culture media for 25 d, then transferring the young cinnamomum camphora 'Yongjin' sprouts in second culture media, cultivating the young cinnamomum camphora 'Yongjin' sprouts in the second culture media for 25 d, transferring the young cinnamomum camphora 'Yongjin' sprouts in propagation culture media and cultivating the young cinnamomum camphora 'Yongjin' sprouts in the propagation culture media for 25 d to completely propagate the cluster buds. The method has the advantages that factors such as basic culture media, hormone, additional substances and culture conditions are inspected, accordingly, problems of weak growth vigor, yellow leaves and browning of 'Yongjin' cluster buds in secondary culture rooms can be effectively solved, and the propagation efficiency can be improved.
Description
Technical field
The invention belongs to biological self reproducing technical field, specifically, relate to a kind of Radix Cinnamomi porrecti and gush gold adventitious buds proliferation method.
Background technology
Lignum cinnamomi camphorae (Cinnamomum caphora) has another name called Radix Cinnamomi porrecti, for the subtropical evergreen broad-leaved arbor of Lauraceae Cinnamomum.Rapidly, flourishing, the four seasons are evergreen in its growth, leaf beautiful and distribute giving off a strong fragrance, are precious fragrant oils seeds and view that city is important and green tree species.Radix Cinnamomi porrecti ' gushing gold ' (Cinnamomum camphora ' Yongjin ') is Radix Cinnamomi porrecti seed variation kind, branch cerise, young leaves, flower are in golden yellow, peel is yellow, and branch, leaf, pericarp color have Seasonal dynamics change, as tall and big color leaf arbor, there is the highest ornamental plantation and popularization and application potentiality and value.But ' gushing gold ' seed hereditary material is unstable, nursery offspring's degradation ratio is up to more than 99%, and kind preserves and breeding difficulty is bigger.At present for obtaining superior clone, carry out Grafting experiments and kept the filial generation of maternal merit to acquisition, yet with elite stand branch limited amount, even if cottage propagation also is difficult to meet the demand of reality.
Plant tissue culture is the important channel of Fast-propagation elite germplasm, tissue culture technique can keep maternal excellent inherited character, the breeding nursery stock of neat and consistent can be provided again in a short time, and can also lay the first stone for seed improvement utilization and genetic transformation, the most in many botanical seedling culturings, obtain application.
Currently acquired Radix Cinnamomi porrecti ' gushing gold ' although tissue culture regeneration technology the most at home and abroad possess some special knowledge, but plant is along with the increase of subculture number, Multiple Buds breeding coefficient reduces, the phenomenons such as growth potential weak, Huang Ye, brownization occur, propagation efficiency is substantially reduced, cannot the acquisition Multiple Buds of stability and high efficiency, and then be difficult to the purpose of Fast-propagation.For realizing the factorial praluction of Radix Cinnamomi porrecti ' gushing gold ' test tube Seedling, the propagation of systematic study Multiple Buds is the most necessary.
Summary of the invention
In view of this, the present invention to solve existing Radix Cinnamomi porrecti and gush the tissue culture regeneration plant of gold along with the increase of subculture number, the problem that propagation efficiency is substantially reduced, it is provided that a kind of Radix Cinnamomi porrecti gushes gold adventitious buds proliferation method.
In order to solve above-mentioned technical problem, the invention discloses a kind of Radix Cinnamomi porrecti and gush gold adventitious buds proliferation method, comprise the following steps: take the Radix Cinnamomi porrecti that axillalry bud not yet sprouts and gush gold young sprout, after aseptic process, be inoculated in inducing culture formation Multiple Buds；The Radix Cinnamomi porrecti forming Multiple Buds is gushed gold young sprout and transfers to proliferated culture medium is cultivated 25d, be then transferred to the second culture medium culturing 25d, finally transfer to proliferated culture medium and cultivate 25d, complete adventitious buds proliferation.
Further, described proliferated culture medium includes MS culture medium, potassium dihydrogen phosphate (KH2PO4), 6-benzyl aminoadenine (6-BA) and heteroauxing (IAA).
Further, KH in described proliferated culture medium2PO4Concentration be 50mg/L, 6-BA concentration be 3mg/L, IAA concentration be 0.02mg/L.
Further, described proliferated culture medium also includes that sucrose, described sucrose concentration are 20-25g/L.
Further, described proliferated culture medium pH value is 5.8-6.0.
Further, described second culture medium includes MS culture medium, KH2PO4, 6-BA and IAA.
Further, KH in described second culture medium2PO4Concentration be 50mg/L, 6-BA concentration be 1mg/L, IAA concentration be 0.02mg/L.
Further, described inducing culture is for including MS culture medium, 6-BA and indolebutyric acid (IBA).
Further, in described inducing culture 6-BA concentration be 1.0mg/L, IBA concentration be 0.02mg/L.
Compared with prior art, the present invention can obtain and include techniques below effect:
1) method of the present invention is by minimal medium, the investigation of the factors such as hormone, added substance, condition of culture, effectively improves Radix Cinnamomi porrecti and gushes that gold successive transfer culture room Multiple Buds growing way is weak, the problem of Huang Ye, brownization, improves propagation efficiency.
Certainly, the arbitrary product implementing the present invention must be not necessarily required to reach all the above technique effect simultaneously.
Accompanying drawing explanation
Accompanying drawing described herein is used for providing further understanding of the present application, constitutes the part of the application, and the schematic description and description of the application is used for explaining the application, is not intended that the improper restriction to the application.In the accompanying drawings:
Fig. 1 is the adventitious buds proliferation figure of culture medium A 1 in the embodiment of the present application；
Fig. 2 is the adventitious buds proliferation figure of culture medium A 2 in the embodiment of the present application；
Fig. 3 is the adventitious buds proliferation figure of culture medium A 3 in the embodiment of the present application；
Fig. 4 is the adventitious buds proliferation figure of culture medium A 4 in the embodiment of the present application；
Fig. 5 is the adventitious buds proliferation figure of culture medium A 5 in the embodiment of the present application；
Fig. 6 is the adventitious buds proliferation figure of culture medium A 6 in the embodiment of the present application；
Fig. 7 is the adventitious buds proliferation figure of culture medium A 7 in the embodiment of the present application；
Fig. 8 is the adventitious buds proliferation figure of culture medium A 8 in the embodiment of the present application；
Fig. 9 is the adventitious buds proliferation figure of culture medium A 9 in the embodiment of the present application；
Figure 10 is the adventitious buds proliferation figure of culture medium B1 in the embodiment of the present application；
Figure 11 is the adventitious buds proliferation figure of culture medium B2 in the embodiment of the present application；
Figure 12 is the adventitious buds proliferation figure of culture medium B3 in the embodiment of the present application；
Figure 13 is the adventitious buds proliferation figure of culture medium B4 in the embodiment of the present application；
Figure 14 is the adventitious buds proliferation figure of culture medium B5 in the embodiment of the present application；
Figure 15 is the adventitious buds proliferation figure of culture medium B6 in the embodiment of the present application.
Detailed description of the invention
Describe embodiments of the present invention in detail below in conjunction with drawings and Examples, thereby the present invention how application technology means are solved technical problem and the technology effect of reaching realizes process and can fully understand and implement according to this.
Embodiment
One, material
Radix Cinnamomi porrecti ' gushing gold ' stem with bud is collected on age period Radix Cinnamomi porrecti ' gushing gold ' tree of Qiu Ai base 14, Ningbo Forest Bureau's seedling center, the young sprout that clip axillalry bud is not yet sprouted takes back laboratory, is seeded in inducing culture (MS+6-BA 1.0mg/L+IBA 0.02mg/L) be formed Multiple Buds after aseptic process.
Two, data process
Proliferation times=25d total bud number/inoculation bud number (height be more than 1cm be effective bud)；Data acquisition meansigma methods ± standard error (SE) represents, use SPSS17.0 statistical software that data are added up, average ratio relatively uses the one-factor analysis of variance, the Different treatments significance difference opposite sex uses Duncan ' s multiple range test method (SSR, P ＜ 0.05).
Three, the propagation of Radix Cinnamomi porrecti ' gushing gold ' Multiple Buds
3.1 different minimal mediums, plant growth regulator kind and the concentration impact on adventitious buds proliferation
For filtering out the culture medium that applicable Radix Cinnamomi porrecti ' gushing gold ' Multiple Buds is constantly bred, the selected three kinds of different minimal mediums of research: the KH of MS, MS and 50mg/L2PO4、1/2MS；0.5,1,3mg/L 6-BA concentration is designed as:；Auxin be IAA:0,0.02,0.04mg/L；GA30,0.05,0.1mg/L (gibberellins):, uses orthogonal scheme to be optimized experiment, and culture medium design is shown in Table 1.
The design of table 1 Radix Cinnamomi porrecti ' gushing gold ' adventitious buds proliferation culture medium
| Numbering | Minimal medium | 6-BA mg/L | IAA mg/L | GA3 mg/L |
| A1 | MS | 0.5 | 0 | 0 |
| A2 | MS | 1 | 0.02 | 0.05 |
| A3 | MS | 3 | 0.04 | 0.1 |
| A4 | The KH of MS+50mg/L2PO4 | 0.5 | 0.02 | 0.1 |
| A5 | The KH of MS+50mg/L2PO4 | 1 | 0.04 | 0 |
| A6 | The KH of MS+50mg/L2PO4 | 3 | 0 | 0.05 |
| A7 | 1/2MS | 0.5 | 0.04 | 0.05 |
| A8 | 1/2MS | 1 | 0 | 0.1 |
| A9 | 1/2MS | 3 | 0.02 | 0 |
Taking Radix Cinnamomi porrecti ' gushing gold ' tissue forming Multiple Buds in inducing culture, put enrichment culture in the different proliferated culture mediums of above-mentioned A1-A9, cultivation results is shown in Fig. 1-Fig. 9 and Biao 2.
The impact on Radix Cinnamomi porrecti ' gushing gold ' adventitious buds proliferation of table 2 different culture media
Note: represent significant difference under 0.05 level with lower cases different after column data.
As shown in figs 1-9, found by the research of Radix Cinnamomi porrecti ' gushing gold ' adventitious buds proliferation, Multiple Buds has a small amount of bud point to be formed in the culture medium (A1, A6, A8) of only 6-BA, but growth difficulty, and in A1 culture medium the yellow leaf of Multiple Buds, base portion brownization is serious, and along with the prolongation leaf abscission of incubation time, plant is dead (such as Fig. 1).And add appropriate auxin and Multiple Buds growth can be made vigorous, leaf color is light green, and its multiplication capacity strengthens along with the rising of auxin.If the basic element of cell division is suitable with the ratio of auxin in culture medium, then the proliferation times of Radix Cinnamomi porrecti ' gushing gold ' Multiple Buds increases rapidly (A3, A4, A5).
The IAA adding suitable concentration in proliferated culture medium is good to the cultivation effect of Radix Cinnamomi porrecti ' gushing gold ' Multiple Buds, and Seedling growth is vigorous, and leaf color is light green.When IAA concentration is 0.04mg/L, propagation being produced inhibitory action, Seedling base portion forms bigger callus, produces a circle brown inhibiting substances and causes base portion leaf to redden, and dried and withered hairs then affects the growth (A3, A5, A7) of whole strain.
GA3Growth and cultivation effect to Radix Cinnamomi porrecti ' gushing gold ' Multiple Buds are without obvious facilitation.GA3Concentration is followed successively by 0,0.05,0.1mg/L, the upgrowth situation of Multiple Buds is not because GA3The rising of concentration and change, and along with GA3Leaf burnt hair, the bud point of formation can not be grown, almost without propagation.
Experimental result also shows, the growth of Radix Cinnamomi porrecti ' gushing gold ' Multiple Buds is had a major impact by different minimal mediums: from culture medium result, with the addition of 50mg/L KH2PO4MS minimal medium and 1/2MS in, during adventitious buds proliferation, brownization has clear improvement (A4, A5, A6).
The impact on Radix Cinnamomi porrecti ' gushing gold ' adventitious buds proliferation effect of 3.2 different sucrose
On the basis of above-mentioned experiment, study the different sucrose impact on Radix Cinnamomi porrecti ' gushing gold ' adventitious buds proliferation effect.Sucrose concentration level is respectively as follows: 10,20,30,40,50,60g/L.Culture medium is the KH of MS, 50mg/L2PO4, the IAA of 6-BA and 0.02mg/L of 2mg/L, agar concentration is 7g/L, pH value 5.8.Experimental result is shown in Figure 10-Figure 15 and Biao 3.
The impact on Radix Cinnamomi porrecti ' gushing gold ' adventitious buds proliferation of table 3 sucrose concentration
Note: represent significant difference under 0.05 level with lower cases different after column data.
As shown in fig. 10-15, when plant tissue culture, owing to the photosynthetic capacity of culture is more weak, so needing to add some saccharides in the medium, on the one hand as the carbon source needed for growth promoter and the energy, certain osmotic pressure can also be maintained simultaneously.In Radix Cinnamomi porrecti ' gushing gold ' adventitious buds proliferation is cultivated, adding of different sucrose amounts all produces certain impact to propagation and growth conditions.It is thin and delicate that Multiple Buds shows as leaf pale green, bud in sucrose medium B1 of low concentration, and also occurs withered and yellow near the blade of base portion；In B3 culture medium, Seedling growth is the most vigorous, and leaf color is light green, does not substantially have withered Seedling to occur；And on the relatively B4 of high-sucrose concentration, B5 medium, Seedling poor growth, differentiation is few, and leaf is yellowish green, thicker and aging, and base portion assembles brown material, has withered Seedling to be entrained in the middle of bud clump.As can be seen from Table 3, along with the increasing of sucrose concentration, growth coefficient the most first raises and then reduces.Simultaneously by table 3 it is recognised that in B2 and B3 culture medium sucrose concentration be 20,30g/L, proliferation times at this moment is maximum, reach 2.80 and 3.38, but because B3 growth coefficient is higher, bud is the thinnest and the most delicate, general production considers more effectively Seedling, so sucrose concentration selects 20～25g/L to be advisable in culture medium.
The 3.3 different pH value impacts on Radix Cinnamomi porrecti ' gushing gold ' adventitious buds proliferation
PH value affects the enrichment culture of Multiple Buds, therefore the following pH value of this experimental design, and to select optimal condition of culture, culture medium includes the KH of MS, 50mg/L2PO4, the IAA of 6-BA and 0.02mg/L of 2mg/L, agar concentration is 7g/L.Experimental result is shown in Table 4.
The different pH value impact on Radix Cinnamomi porrecti ' gushing gold ' adventitious buds proliferation of table 4
Note: represent significant difference under 0.05 level with lower cases different after column data.
PH in culture medium value is also the key factor affecting plant tissue culture.The change of pH value has influence on the dissolubility of some ions and has influence on plant and the absorption of each element is occurred nutritional deficiency symptom.The most too high or too low pH value influences whether the solidification of culture medium, and pH value is bigger than normal, culture medium can be made to harden, otherwise then culture medium is too soft.PH value is when 5.5, and sterilizing wild Oryza species is softer, and bud-leaf of growing thickly is grayish green, and the Multiple Buds of differentiation has vitrification trend and bud poor growth；PH value is in the culture medium of 5.8～6.5, and Radix Cinnamomi porrecti ' gushing gold ' Multiple Buds all can grow more normally, and proliferation times is high, and effective bud is many；When pH value is higher than 7.0, bud-leaf of growing thickly shows yellowish green situation, and bud poor growth, and proliferation times reduces.As shown in Table 4, Radix Cinnamomi porrecti ' gushing gold ' Multiple Buds when pH value is between 5.8～6.0, sprout can vigorous growth, and higher proliferation times can be maintained.
The 3.4 different subculture mode impacts on Radix Cinnamomi porrecti ' gushing gold ' adventitious buds proliferation
Following 5 kinds of culture medium are carried out alternate culture, and cultivation results is shown in Table 5.
1. the KH of MS, 50mg/L2PO4, the IAA of 6-BA and 0.02mg/L of 3mg/L；
2. the KH of MS, 50mg/L2PO4, the IAA of 6-BA and 0.02mg/L of 2mg/L；
3. the KH of MS, 50mg/L2PO4, the IAA of 6-BA and 0.02mg/L of 1mg/L；
4. the KH of MS, 50mg/L2PO4, the IAA of 6-BA and 0.02mg/L of 0.5mg/L；
5. the KH of MS, 50mg/L2PO4IAA with 0.02mg/L.
The impact on Radix Cinnamomi porrecti ' gushing gold ' adventitious buds proliferation of the table 5 not subculture mode
Note: represent significant difference under 0.05 level with lower cases different after column data.
The tissue cultured seedling propagation typically continuous subculture of outer implant is in containing in 6-BA proliferated culture medium, and this mode frequently results in vitrification or the growth deformity of tissue cultured seedling, therefore compares different subculture mode to Radix Cinnamomi porrecti ' gushing gold ' adventitious buds proliferation and the impact of growth.First outer implant is inoculated in the KH of the proliferated culture medium MS+50mg/L of filtered out Multiple Buds2PO4In the IAA (1.) of the 6-BA+0.02mg/L of+3mg/L, it is transferred to (2., 3., 4.) that concentration is gradually lowered after 25d respectively or removes in 6-BA culture medium (5.), transferring to after 25d in proliferated culture medium in 1..Result shows (table 5), and in five kinds of subculture modes, D3 mode is most beneficial for the propagation of Radix Cinnamomi porrecti ' gushing gold ' Multiple Buds, and average proliferation coefficient is the highest, and blastogenesis length is quick；Growth coefficient can be caused substantially to reduce in culture medium 1. middle cultivation continuously, and vitrification easily occurs in Multiple Buds.
Radix Cinnamomi porrecti ' gushing gold ' is grown thickly the impact of propagation by 3.5 different subculture cycles
Experiment finding, subculture cycle is relatively big on the impact of Radix Cinnamomi porrecti ' gushing gold ' adventitious buds proliferation, so designing following experimental program, to filter out suitable Subculture Time, the KH of culture medium MS+50mg/L2PO4The IAA of the 6-BA+0.02mg/L of+2mg/L, agar concentration is 7g/L, and pH value 5.8, experimental result is shown in Table 6.
Table 6 is with continuing for the cycle impact on Radix Cinnamomi porrecti ' gushing gold ' adventitious buds proliferation
Note: represent significant difference under 0.05 level with lower cases different after column data.
In incubation, material does not shifts, and can cause culture materials brown stain, and final material is the most dead.The culture medium that upgrades in time is one of effectively important measures reducing brownization.For the proliferative properties of Radix Cinnamomi porrecti ' gushing gold ' Multiple Buds, compare the impact that it is grown by different subculture cycle.Result shows, it is minimum that cultivation 10d i.e. shifts proliferation times, and its proliferation times occurs first increasing the trend reduced afterwards with extending of subculture cycle.Subculture cycle compares in significant difference (p < 0.05) at 25d～30d proliferation times with other process, and subculture cycle its proliferation times of 25d averagely up to.Although the proliferation times of 30d subculture Multiple Buds is the highest, but Multiple Buds growing state starts to deteriorate, and yellow leaf and the phenomenon of dead Seedling occurs, is unfavorable for continuing to cultivate.Therefore 25～30d are selected to carry out subculture and be preferred, beneficially the growth of Multiple Buds and propagation.
Four, Radix Cinnamomi porrecti ' gushing gold ' the adventitious buds proliferation method finally determined
Take Radix Cinnamomi porrecti ' gushing gold ' young sprout that axillalry bud is not yet sprouted, be inoculated in inducing culture (MS+6-BA 1.0mg/L+IBA 0.02mg/L) after aseptic process and form Multiple Buds；Radix Cinnamomi porrecti ' gushing gold ' young sprout forming Multiple Buds is transferred to proliferated culture medium (MS+KH2PO450mg/L+6-BA 3.0mg/L+IAA 0.02mg/L+ sucrose 20-25g/L, pH=5.8-6.0) cultivate 25d, it is then transferred to the second culture medium (MS+KH2PO450mg/L+6-BA 1.0mg/L+IAA 0.02mg/L) cultivate 25d, finally transfer to proliferated culture medium (MS+KH2PO450mg/L+6-BA 3.0mg/L+IAA 0.02mg/L+ sucrose 20-25g/L, pH=5.8-6.0) cultivate 25d, complete adventitious buds proliferation.
The method of the present invention by minimal medium, the investigation of the factors such as hormone, added substance, condition of culture, effectively improves that Radix Cinnamomi porrecti ' gushing gold ' successive transfer culture room Multiple Buds growing way is weak, the problem of Huang Ye, brownization, improves propagation efficiency.
As employed some vocabulary in the middle of description and claim to censure special component or method.Those skilled in the art are it is to be appreciated that same composition may be called with different nouns in different regions.In the way of this specification and claims not difference by title is used as distinguishing composition." comprising " as mentioned by the middle of description and claim in the whole text is an open language, therefore should be construed to " comprise but be not limited to "." substantially " referring in receivable range of error, those skilled in the art can solve described technical problem in the range of certain error, basically reaches described technique effect.Description subsequent descriptions is to implement the better embodiment of the present invention, for the purpose of right described description is the rule so that the present invention to be described, is not limited to the scope of the present invention.Protection scope of the present invention is when being as the criterion depending on the defined person of claims.
It can further be stated that, term " includes ", " comprising " or its any other variant are intended to comprising of nonexcludability, so that include that the commodity of a series of key element or system not only include those key elements, but also include other key elements being not expressly set out, or also include the key element intrinsic for this commodity or system.In the case of there is no more restriction, statement " including ... " key element limited, it is not excluded that there is also other identical element in the commodity including described key element or system.
Described above illustrate and describes some preferred embodiments of the present invention, but as previously mentioned, it is to be understood that the present invention is not limited to form disclosed herein, it is not to be taken as the eliminating to other embodiments, and can be used for other combinations various, amendment and environment, and can be modified by above-mentioned teaching or the technology of association area or knowledge in invention contemplated scope described herein.And the change that those skilled in the art are carried out and change are without departing from the spirit and scope of the present invention, the most all should be in the protection domain of claims of the present invention.
Claims (9)
1. Radix Cinnamomi porrecti gushes gold adventitious buds proliferation method, it is characterised in that comprise the following steps: takes the Radix Cinnamomi porrecti that axillalry bud not yet sprouts and gushes gold young sprout, is inoculated in inducing culture formation Multiple Buds after aseptic process；The Radix Cinnamomi porrecti forming Multiple Buds gushes gold young sprout transfer to cultivate 25d during proliferated culture medium is cultivated, be then transferred to the second culture medium culturing 25d, finally transfer to proliferated culture medium and cultivate 25d, complete adventitious buds proliferation.
2. Radix Cinnamomi porrecti as claimed in claim 1 gushes gold adventitious buds proliferation method, it is characterised in that described proliferated culture medium includes MS culture medium, KH2PO4, 6-BA and IAA.
3. Radix Cinnamomi porrecti as claimed in claim 2 gushes gold adventitious buds proliferation method, it is characterised in that KH in described proliferated culture medium2PO4Concentration be 50mg/L, 6-BA concentration be 3mg/L, IAA concentration be 0.02mg/L.
4. Radix Cinnamomi porrecti as claimed in claim 3 gushes gold adventitious buds proliferation method, it is characterised in that described proliferated culture medium also includes that sucrose, described sucrose concentration are 20-25g/L.
5. Radix Cinnamomi porrecti as claimed in claim 4 gushes gold adventitious buds proliferation method, it is characterised in that described proliferated culture medium pH value is 5.8-6.0.
6. Radix Cinnamomi porrecti as claimed in claim 5 gushes gold adventitious buds proliferation method, it is characterised in that described second culture medium includes MS culture medium, KH2PO4, 6-BA and IAA.
7. Radix Cinnamomi porrecti as claimed in claim 6 gushes gold adventitious buds proliferation method, it is characterised in that KH in described second culture medium2PO4Concentration be 50mg/L, 6-BA concentration be 1mg/L, IAA concentration be 0.02mg/L.
8. Radix Cinnamomi porrecti as claimed in claim 7 gushes gold adventitious buds proliferation method, it is characterised in that inducing culture described in described inducing culture is for including MS culture medium, 6-BA and IBA.
9. Radix Cinnamomi porrecti as claimed in claim 8 gushes gold adventitious buds proliferation method, it is characterised in that in described inducing culture 6-BA concentration be 1.0mg/L, IBA concentration be 0.02mg/L.
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