CN108812312A - A kind of induction of cryptomeria callus from stem segment and plant regeneration cultural method - Google Patents
A kind of induction of cryptomeria callus from stem segment and plant regeneration cultural method Download PDFInfo
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- CN108812312A CN108812312A CN201810600276.1A CN201810600276A CN108812312A CN 108812312 A CN108812312 A CN 108812312A CN 201810600276 A CN201810600276 A CN 201810600276A CN 108812312 A CN108812312 A CN 108812312A
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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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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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Abstract
The invention discloses a kind of cryptomeria stem section explant callus induction and plant regeneration methods, and this approach includes the following steps：Selection cryptomeria tissue culture seeding stem segment is tissue cultures material, and tissue culture seeding stem segment is linked into callus inducing medium；After callus induction comes out, callus is linked into the induction that adventitious bud is carried out on differential medium；Adventitious bud inducing is transferred to DCR and cultivates the growth for carrying out adventitious bud and adventitious root substantially after coming out.Compared with cryptomeria tissue culture method in the prior art, cryptomeria tissue culture seeding stem segment regenerated plant culture method advantage of the invention is：It efficiently solves the problems, such as that cryptomeria explant, by callus approach plant regeneration, realizes the tissue culture system of cryptomeria stability and high efficiency under conditions of tissue culture, provides technical support to accelerate cryptomeria tissue-cultured seedling plant regeneration.
Description
Technical field
The invention belongs to plant tissue culture technical fields, and in particular to a kind of induction of cryptomeria callus from stem segment and plant regeneration
Cultural method.
Background technique
There are two types of cryptomeria (Cryptomeria fortunei) is total in worldwide, most of plant classification scholar recognizes
It is divided into cryptomeria (C.fortunei) and Japanese cedar (C.japonica) for Cryptomeria.Cryptomeria is tree species specific to China, is produced
In the ground such as Zhejiang, the Fujian and Jiangxi area below height above sea level 1100m.In South of Jiangsu Province, Zhejiang, Anhui, Henan, Hubei, Guangxi
And there are cultivation, and well-grown in the ground such as Guangdong.The cryptomeria greening coniferous species important as south China, trunk is tall and big,
Material is light and soft, and texture is straight, and intensity is stronger, and contract with dry rate is small, and easily dry, corrosion resistance is stronger, with each side such as material and environmental protections
Face suffers from outstanding performance, and mainly has stronger resistance to acid rain in terms of environmental protection, to the purification of air, bear from
Sub- effect etc. has notable contribution, and cryptomeria essential oil also has poisoning to black chest termite.
Domestic market is larger to cryptomeria demand, however cryptomeria mainly uses conventional mating system in China, right
Cannot saving well for some merits, plays and applies.Therefore, the research for carrying out the pierre of Cryptomeria is that have very much
It is necessary.
Summary of the invention
Goal of the invention：For the deficiencies in the prior art, the object of the present invention is to provide a kind of cryptomeria stem section callus
Tissue induction and plant regeneration cultural method carry out callus using the stem section of cryptomeria choiceness as tissue cultures material
Induction and plant regeneration, provide the support of theory and technology for the proliferation of propagation of cryptomeria breeding.
Technical solution：In order to achieve the above-mentioned object of the invention, the technical solution adopted by the present invention is：
A kind of induction of cryptomeria callus from stem segment and plant regeneration cultural method, include the following steps：
1) stem section for choosing cryptomeria choiceness tissue-cultured seedling is tissue cultures material, and the needle of stem section is wiped out, take 3~
The stem section of 4cm long accesses callus inducing medium culture, makes well differentiated stem section dedifferentiation callus, wherein
Callus inducing medium formula is：DCR+2 is improved, 4-D 0.5-2.0mg/L+6-BA 0.1-1.5mg/L+ hydrolyzes junket egg
White 400-600mg/L+L- glutamine 400-600mg/L+30-40g/L sucrose；
2) callus access subculture medium is subjected to shoot proliferation culture；The subculture of cryptomeria stem section evoked callus
Proliferation culture medium formula is improvement DCR+2,4-D 0.5-2.0mg/L+6-BA 0.1-1.5mg/L+25-35g/L sucrose；
3) callus that cryptomeria stem section induces is transferred to adventitious bud induction culture base, Fiber differentiation generates adventitious bud；No
Normal bud induced medium is：Improve DCR+2,4-D 1.0mg/L+6-BA 1.2mg/L；
4) it when adventitious bud grows to 2-3cm, is cut from callus, it is indefinite that adventitious bud is transferred to growth medium progress
The grown cultures of bud and adventitious root, growth medium are DCR minimal medium.
In step 1), callus inducing medium formula is：Improve DCR+2,4-D 1.0mg/L+6-BA 0.5mg/L+
Caseinhydrolysate 500mg/L+L- glutamine 400mg/L+40g/L sucrose.
In step 1), cryptomeria choiceness is No. 011 clone, and condition of culture is 25 DEG C, dark culture.
In step 2), subculture multiplication medium formula is：Improve DCR+2,4-D 1.0mg/L+6-BA 0.5mg/L+30g/
L sucrose.
In step 2), callus is in improvement DCR+0.5mg/L 6-BA+1.0mg/L 2,4-D, improvement DCR+0.5mg/L
Two kinds of culture mediums of 6-BA+2.0mg/L 2,4-D alternately squamous subculture, callus are able to maintain preferable state and proliferation speed
Degree.
In step 2), 20d subculture is primary.
In step 3), adventitious bud induction culture base is：Improve DCR+2,4-D 1.0mg/L+6-BA 1.2mg/L+1.0mg/
LKT+0.05mg/LNAA。
In step 4), 28d successfully grows new adventitious bud and adventitious root on DCR minimal medium, after taking root, carries out normal
Rule culture, can plant regeneration and cultivate seedling.
Beneficial effect：Compared with prior art, cryptomeria callus from stem segment of the invention induction and plant regeneration culture side
Method efficiently solves the problems, such as that cryptomeria explant, by callus approach plant regeneration, realizes willow under conditions of tissue culture
The tissue culture system of China fir stability and high efficiency provides technical support to accelerate cryptomeria tissue-cultured seedling plant regeneration.
Detailed description of the invention
Fig. 1 is state diagram of the different clones on calli induction media；In figure, A：No. 59 callus induction shapes
State, B：The callus that 011 extra implant induces；
Fig. 2 is subculture state diagram of the callus in different culture medium；In figure, A：Callus is on optimal medium
The state of subculture, 20 under microscope × lower observation；B：The callus state of subculture on the induction medium for a long time, under microscope
12.5 × observation down；
Fig. 3 is tissue culture seeding stem segment callus differentiation adventitious bud procedure chart；In figure, A：There is blade in callus, micro-
16 under mirror × observation；B：Adventitious bud, 10 × observation under microscope are grown on callus；C：Complete adventitious bud, under the microscope
10 × observation；D：Adventitious bud is grown completely, 12.5 under microscope × observation；
Fig. 4 is that the adventitious bud proliferation of tissue culture seeding stem segment callus and adventitious root generate situation map；In figure, (A：Callus group
Knit the adventitious bud induced cut be transferred to DCR minimal medium carry out adventitious bud proliferation；B：By the culture of 28d, explant is not
Normal bud stretches and adventitious root is grown；
Fig. 5 is addition ABA and inositol proliferation and subculture result figure；In figure, A callus is in ABA optium concentration, B callus group
It is woven in inositol optium concentration.
Specific embodiment
The present invention is described further combined with specific embodiments below.
Embodiment 1
A kind of induction of cryptomeria callus from stem segment and plant regeneration cultural method, specific step is as follows：
1) callus from stem segment induces
The stem section for choosing cryptomeria choiceness 011 (Japan) and No. 59 (Zhejiang stone Yang) tissue-cultured seedling is tissue cultures material
Material, the needle of stem section is wiped out, and is taken the stem section of 3~4cm long to access callus inducing medium culture, is made well differentiated stem
Section dedifferentiation is callus, wherein callus inducing medium formula is：Improve DCR+2,4-D 1.0mg/L+6-BA
0.5mg/L+ caseinhydrolysate (CH) 500mg/L+L- glutamine 400mg/L+ sucrose 4%；Condition of culture is 25 DEG C, dark to train
It supports, generally cultivates stem section dedifferentiation in 15 days or so and form callus.
As shown in Figure 1, No. 011 clone (Figure 1B) Callus induction rate on callus inducing medium is 100%,
It can be seen that callus when 15d, mellow and full glossy, the white slightly transparent, close structure in callus surface, in 25d, callus group
Fabric/color becomes yellowish or pale green, and surface is mellow and full, relatively dry, close structure, graininess protrusion, and growth rate is very fast.No. 59 nothings
Property system (Figure 1A) on callus inducing medium, callus time of occurrence is slower, about 40d or so occur, callus
Inducing amount is small, and the easy browning of explant is dead, and value-added speed is slower, and callus quality is soft, white or faint yellow, surface is dry
Dry, part callus is slightly wet, inviscid.
2) callus squamous subculture
In the present embodiment Subculture, to will appear growth rate slow for long-term subculture on the induction medium for callus
Slowly, graininess protrusion and nadel and the phenomenon that deposit, in order to preferably directly induce adventitious bud, this reality from callus
It applies example and corresponding adjustment has been carried out with the concentration of 6-BA to 2,4-D, test result shows：Callus 2,4-D 2.0mg/L,
When both concentration of 6-BA 0.5mg/L and 2,4-D 1.0mg/L, 6-BA0.5mg/L hand over successive generations, callus can be protected
Preferable state and growth rate are held, callus surface is mellow and full, dry, graininess protrusion, close structure (Fig. 2A).It is inducing for a long time
Callus on culture medium will appear surface wettability, nadel, the soft state of quality (Fig. 2 B).Therefore, comprehensively consider
Afterwards, the subculture medium of the callus of No. 011 clonal tissue culture seedling stem section induction should be chosen：Improve DCR+0.5mg/L 6-BA
+ 1.0mg/L 2,4-D, improvement DCR+0.5mg/L 6-BA+2.0mg/L 2, two kinds of culture mediums of 4-D alternately squamous subculture,
Callus status can be made to keep best, growth rate is also most fast state.
Suitable subculture cycle has vital effect to the holding of callus and proliferation, while can also be effective
Inhibit the browning of callus.Result of this example indicate that：20d subculture is once optimal subculture cycle.Specific test
It the results are shown in Table 1.
Influence of the different Subculture Times of table 1 to callus
3) by callus induction adventitious bud
The callus that cryptomeria stem section induces is transferred to adventitious bud induction culture base, callus is in adventitious bud induction culture
Adventitious bud can be generated by growth in one month on base；The callus adventitious bud induction culture that cryptomeria tissue culture seeding stem segment induces
Base is：DCR+2 is improved, 4-D1.0mg/L+6-BA 1.2mg/L, callus status is best at this time；Callus gradually greening,
Dry tack free, there is graininess protrusion, and structure is compact.
On the medium base of improvement DCR culture medium+1.2mg/L6-BA+1.0mg/L2,4-D, 0.05mg/ is added
LTDZ+2.0mg/LKT+0.05mg/LNAA, it is in the best state, but callus is easy browning；Add 1.0mg/LKT+0.05mg/
LNAA, callus state is preferable, and browning degree mitigates, and successfully induces adventitious bud, and Induction Process is as shown in Figure 3.
As shown in Figure 3：1.2mg/L6-BA and 1.0mg/L2 is added on improvement DCR culture medium, when 4-D is broken up, more
Injured tissue starts the blade (Fig. 3 A) for adventitious bud occur, but can not induce complete adventitious bud always on the culture medium, therefore,
When adding KT and NAA of suitable concentration on the culture medium, adventitious bud starts (Fig. 3 B, C) occur on callus, KT
Optium concentration be 2.0mg/L, NAA optium concentration be 0.05mg/L；On differential medium after subculture 2 times, adventitious bud is complete
Grow (Fig. 3 D).
4) adventitious bud that tissue culture seeding stem segment callus induction goes out is cut, is transferred to DCR minimal medium and is grown,
New adventitious bud and adventitious root (Fig. 4 A, B) are successfully grown on 28d or so, DCR minimal medium.After taking root, conventional training is carried out
Support, can plant regeneration and cultivate seedling.
Embodiment 2
A kind of induction of cryptomeria callus from stem segment and plant regeneration cultural method, specific step is as follows：
1) callus from stem segment induces
The stem section for choosing No. 011 tissue-cultured seedling of cryptomeria choiceness is tissue cultures material, and the needle of stem section is wiped out, is taken
The stem section of 3~4cm long accesses callus inducing medium culture, makes well differentiated stem section dedifferentiation callus,
In, callus inducing medium formula is：Minimal medium+2,4-D 1.0mg/L+6-BA 0.5mg/L+ caseinhydrolysate
500mg/L+L- glutamine 400mg/L+ sucrose 4%；Condition of culture is 25 DEG C, dark culture, the every 18-20 days subcultures of culture materials
Culture is primary, generally cultivates stem section dedifferentiation in 15 days or so and forms callus.Different minimal mediums are cured cryptomeria tissue-cultured seedling
The results are shown in Table 1 for wound induction.
The influence that the different minimal mediums of table 1 induce cryptomeria tissue-cultured seedling callus
As it can be seen from table 1 the explant on WPM culture medium, when 25d, only a small amount of callus induction goes out
Come, DCR and the inductivity for improveing DCR and appreciation rate more preferably compared with other three kinds of culture medium inductivities and appreciation rate, but improve DCR
The callus speed of growth and state ratio DCR in more preferably.
2) callus proliferation
When the callus of cryptomeria tissue-cultured seedling No. 011 stem section of clone is induced out and covers explant, by callus
Tissue is transferred to proliferated culture medium culture, and proliferation culture medium formula is：Improve DCR+2,4-D1.0mg/L+6-BA 0.5mg/L+
30g/L sucrose adds ABA and inositol on this basis.
The result shows that：When ABA concentration is 4.0mg/L, new callus (figure is grown on the callus of browning
5A)；ABA concentration is too low, cannot effectively promote the proliferation of callus；ABA excessive concentration can then inhibit the increasing of callus
It grows, and callus quality is soft, starts nadel occur；But the ABA concentration of either which kind of concentration can all make callus
It organizes quality soft, nadel occurs, being unfavorable for the later period, directly induction differentiates adventitious bud from callus.Inositol concentration
When for 2.0g/L, callus dry tack free, mellow and full, graininess protrusion, close structure (Fig. 5 B)；Inositol concentration is too low,
Callus proliferation rate is lower, and the easy browning of callus；Inositol concentration is excessively high, then can inhibit the multiplication rate of callus, and
Callus hydration phenomena is serious, there is nadel.
Callus in induced medium is inoculated into the sucrose culture medium for being added to 3 kinds of various concentrations, test knot
Fruit discovery：When sucrose concentration is 30g/L, callus status is best, and multiplication rate is also most fast.Sucrose concentration is
When 20g/L, callus quality is relatively soft, and white is slightly transparent, and growth rate is slower；When sucrose concentration is 40g/L,
Callus quality is harder, easily loose, and white is partially yellow, and growth rate is slower.
3) by callus induction adventitious bud
The callus that cryptomeria stem section induces is transferred to adventitious bud induction culture base, callus is in adventitious bud induction culture
Adventitious bud can be generated by growth in one month on base；The callus adventitious bud induction culture that cryptomeria tissue culture seeding stem segment induces
Base is：Improve DCR+2,4-D 1.0mg/L+6-BA 1.2mg/L+KT 1.0mg/L+NAA 0.05mg/L+30g/L sucrose.
4) adventitious bud that tissue culture seeding stem segment callus induction goes out is cut, is transferred to DCR minimal medium and is grown, is added
Add 30g/L sucrose, 16h illumination/8h dark culture.After a squamous subculture, adventitious bud proliferation, adventitious root is grown.After taking root,
Carry out routine culture, can plant regeneration and cultivate seedling.
Claims (8)
1. a kind of cryptomeria callus from stem segment induction and plant regeneration cultural method, which is characterized in that include the following steps：
1) stem section for choosing cryptomeria choiceness tissue-cultured seedling is tissue cultures material, and the needle of stem section is wiped out, 3~4cm is taken
Long stem section accesses callus inducing medium culture, makes well differentiated stem section dedifferentiation callus, wherein callus
Tissue Fiber differentiation based formulas be：Improve DCR+2,4-D 0.5-2.0mg/L+6-BA 0.1-1.5mg/L+ caseinhydrolysate
400-600mg/L+L- glutamine 400-600mg/L+30-40g/L sucrose；
2) callus access subculture medium is subjected to shoot proliferation culture；The shoot proliferation of cryptomeria stem section evoked callus
Culture medium prescription is improvement DCR+2,4-D 0.5-2.0mg/L+6-BA 0.1-1.5mg/L+25-35g/L sucrose；
3) callus that cryptomeria stem section induces is transferred to adventitious bud induction culture base, Fiber differentiation generates adventitious bud；Adventitious bud
Induced medium is：Improve DCR+2,4-D 1.0mg/L+6-BA 1.2mg/L；
4) when adventitious bud grows to 2-3cm, cut from callus, by adventitious bud be transferred to growth medium carry out adventitious bud with
The grown cultures of adventitious root, growth medium are DCR minimal medium.
2. cryptomeria callus from stem segment induction according to claim 1 and plant regeneration cultural method, it is characterised in that：Step
It is rapid 1) in, callus inducing medium formula is：Improve DCR+2,4-D 1.0mg/L+6-BA 0.5mg/L+ caseinhydrolysate
500mg/L+L- glutamine 400mg/L+40g/L sucrose.
3. cryptomeria callus from stem segment induction according to claim 1 and plant regeneration cultural method, it is characterised in that：Step
It is rapid 1) in, cryptomeria choiceness be No. 011 clone, condition of culture be 25 DEG C, dark culture.
4. cryptomeria callus from stem segment induction according to claim 1 and plant regeneration cultural method, it is characterised in that：Step
It is rapid 2) in, subculture multiplication medium formula is：Improve DCR+2,4-D 1.0mg/L+6-BA 0.5mg/L+30g/L sucrose.
5. cryptomeria callus from stem segment induction according to claim 1 and plant regeneration cultural method, it is characterised in that：Step
It is rapid 2) in, callus improvement DCR+0.5mg/L 6-BA+1.0mg/L 2,4-D, improvement DCR+0.5mg/L 6-BA+
Two kinds of culture mediums of 2.0mg/L 2,4-D alternately squamous subculture, callus are able to maintain preferable state and growth rate.
6. cryptomeria callus from stem segment induction according to claim 1 and plant regeneration cultural method, it is characterised in that：Step
It is rapid 2) in, 20d subculture is primary.
7. cryptomeria callus from stem segment induction according to claim 1 and plant regeneration cultural method, it is characterised in that：Step
It is rapid 3) in, adventitious bud induction culture base is：Improve DCR+2,4-D 1.0mg/L+6-BA 1.2mg/L+1.0mg/LKT+
0.05mg/LNAA。
8. cryptomeria callus from stem segment induction according to claim 1 and plant regeneration cultural method, it is characterised in that：Step
It is rapid 4) in, 28d successfully grows new adventitious bud and adventitious root on DCR minimal medium, after taking root, carries out routine culture
The regeneration and cultivation seedling of plant.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109744148A (en) * | 2019-01-31 | 2019-05-14 | 山东农业大学 | A kind of forest shoot regeneration method based on apical meristem transdifferentiation |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103609442A (en) * | 2013-11-15 | 2014-03-05 | 四川农业大学 | Method for obtaining regenerated plants by tissue culture of stems of cryptomeria fortunei |
| CN104255524A (en) * | 2014-10-15 | 2015-01-07 | 广西大学 | Method for quickly producing Chinese fir seedlings through micro-cutting |
| CN106922534A (en) * | 2017-03-27 | 2017-07-07 | 河池乐康生态农业科技有限公司 | A kind of method that tissue cultures quickly breed China fir |
-
2018
- 2018-06-11 CN CN201810600276.1A patent/CN108812312B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103609442A (en) * | 2013-11-15 | 2014-03-05 | 四川农业大学 | Method for obtaining regenerated plants by tissue culture of stems of cryptomeria fortunei |
| CN104255524A (en) * | 2014-10-15 | 2015-01-07 | 广西大学 | Method for quickly producing Chinese fir seedlings through micro-cutting |
| CN106922534A (en) * | 2017-03-27 | 2017-07-07 | 河池乐康生态农业科技有限公司 | A kind of method that tissue cultures quickly breed China fir |
Non-Patent Citations (1)
| Title |
|---|
| 席梦利等: "杉木合子胚愈伤组织的诱导及植株再生", 《南京林业大学学报(自然科学版)》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109744148A (en) * | 2019-01-31 | 2019-05-14 | 山东农业大学 | A kind of forest shoot regeneration method based on apical meristem transdifferentiation |
| CN109744148B (en) * | 2019-01-31 | 2019-10-29 | 山东农业大学 | A kind of forest shoot regeneration method based on apical meristem transdifferentiation |
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Application publication date: 20181116 Assignee: Huazhilin flower garden of Huai'an Economic and Technological Development Zone Assignor: Nanjing Forestry University Contract record no.: X2019320000285 Denomination of invention: Cryptomeria fortunei stem segment callus induction and plant regeneration culture method Granted publication date: 20190927 License type: Common License Record date: 20191119 Application publication date: 20181116 Assignee: Nanjing Lvzhou Shell Division Pecan Co., Ltd Assignor: Nanjing Forestry University Contract record no.: X2019320000286 Denomination of invention: Cryptomeria fortunei stem segment callus induction and plant regeneration culture method Granted publication date: 20190927 License type: Common License Record date: 20191119 Application publication date: 20181116 Assignee: Huangshan Weiyuan Seedling Development Co., Ltd. Assignor: Nanjing Forestry University Contract record no.: X2019320000284 Denomination of invention: Cryptomeria fortunei stem segment callus induction and plant regeneration culture method Granted publication date: 20190927 License type: Common License Record date: 20191119 |