CN106489731A - A kind of dove tree efficiently induces the tissue culture and rapid propagation method of Multiple Buds - Google Patents
A kind of dove tree efficiently induces the tissue culture and rapid propagation method of Multiple Buds Download PDFInfo
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- CN106489731A CN106489731A CN201610910782.1A CN201610910782A CN106489731A CN 106489731 A CN106489731 A CN 106489731A CN 201610910782 A CN201610910782 A CN 201610910782A CN 106489731 A CN106489731 A CN 106489731A
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- Prior art keywords
- dove tree
- bud
- lateral bud
- dove
- multiple buds
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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
-
- 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
Abstract
The invention discloses a kind of dove tree efficiently induces the tissue culture and rapid propagation method of Multiple Buds, concretely comprise the following steps：1）Growth selection stalwartness, no disease and pests harm, character are excellent, lateral bud is formed and dove tree branch during non-lamina is maternal plant, add and are soaked in GA3 solution, grow into more than 3cm to dove tree lateral bud；2）Clip lateral bud, carries out disinfection, and is then inoculated in Wpm+KT1.2mg L‑1+NAA0.3mg·L‑1+6‑BA0.5 mg·L‑1+ agar 3.5g L‑1+ sucrose 20g L‑1+AC0.2g·L‑1Culture medium in；Bud point germination and growth after 18 22d is cultivated, Multiple Buds are formed.The dove tree inducing clumping bud technology provided using the present invention, method are easy, and induction duration is short, and breeding coefficient is high, and there is certain facilitation to this country-level rare and endangered plant protection.
Description
Technical field
The present invention relates to a kind of plant tissue culture reproduction technique, specially a kind of dove tree efficiently induces the tissue-culturing rapid propagation of Multiple Buds
Method.
Background technology
Dove tree is deciduous tree.15～25 meters high can be grown into, leaf is extensively avette, and there is sawtooth at edge.Dyeing defect is very beautiful, is
The Relict Plant that the 10000000 years Pre-Cenozoic Tertiary Period stayed, in quaternary glacier period, the dove tree of most area is gone out in succession
Absolutely, only survive in some areas of southern china, become " living fossil " of plant kingdom's today, be described as " China
Pigeon tree ", also known as " pigeon flower tree ", " water pear ", wild species are only grown in Sichuan Province China and save and Hubei Province surrounding area.Dove tree
Country-level Top-rated protected wild plants are listed in, have been the distinctive single platymiscium of China, belonged to Relict Plant, and the whole world has write
The ornamental plant of name.
At present, the reproduction technique of dove tree is mainly based on seeding and seedling raising, but this mode has the following disadvantages：The seedling cycle
Long, seedling-growing time is 3-10；Emergence rate is low, irregular, easy variation, has certain impact on the merit for preserving maternal plant；With
When need a large amount of human and material resources and land resource, production cost is high；In sum, quick, the dove tree of low production cost that emerges is found
Tissue-culturing rapid propagation mode, promotes industrialization production that there is necessity.
Southwestern Normal University Zou Li beautiful《Plant research》Deliver《The tissue culture of Endangered Plant Davidia involucrate Baill is with plant again
Raw》The quick-breeding method that a kind of employing dove tree lateral bud directly induces Multiple Buds is disclosed, but the method has explant acquisition time
The limited, shortcoming of induction time length, and its appreciation rate and proliferation times all have a certain upgrade space；Sichuan Agricultural University's summer
Break is delivered《Dove tree initial culture is studied》Deliver with Hubei ethnic university sieve generation honor《Dove tree sprout Study on tissue culture》In
All mentioning with dove tree lateral bud carries out bud induction as explant, but can only all promote simple bud to sprout, it is impossible to induce Multiple Buds.
Content of the invention
It is an object of the invention to provide a kind of dove tree efficiently induces the tissue culture and rapid propagation method of Multiple Buds, which is easy to operate, induction
Speed is fast, and growth coefficient is high.
For solving above-mentioned technical problem, the technical solution adopted in the present invention is：A kind of dove tree efficiently induces Multiple Buds
Tissue culture and rapid propagation method, comprises the following steps：
1) growth selection stalwartness, no disease and pests harm, character be excellent, lateral bud is formed and dove tree branch during non-lamina is maternal plant,
Add and soaked in GA3 solution, more than 3cm is grown into dove tree lateral bud；
2) clip lateral bud, carries out disinfection, and is then inoculated in Wpm+KT1.2mg L-1+NAA0.3mg·L-1+6-
BA0.5mg·L-1+ agar 3.5g L-1+ sucrose 20g L-1+AC0.2g·L-1Culture medium in；After culture 18-22d, bud point is sprouted
Grow, form Multiple Buds.
The time for dove tree branch being acquired and being processed is the March of September then to next year.
Preferably, the time for dove tree branch being acquired and being processed is March.Now will sprout naturally for lateral bud
When, germination rate, the growth of bud length are fast.
Step 1) in dove tree branch be intercept raw then from top to bottom to the 5th lateral bud branch.
Step 1) in immersion when using 100mg/L GA3 solution, changed a solution per 5 days.
Step 2) in the lateral bud of clip contain 1-2mm xylem.
Step 2) in sterilization when, first flowing water rinses 1h, using 0.4% detergent immersion 4-6min of mass concentration, then adopts
With 75% alcohol-pickled 20s of volumetric concentration, aseptic water washing 2 times, then use mass concentration 0.1%Hgcl2Immersion 3-5min, finally
Aseptic water washing 4 times.
The environment of culture is 25 ± 1 DEG C of temperature, humidity 75 ± 2%, illumination 12-14h/d, light intensity 1800-2200lx.
Step 1) in immersion process processed in outdoor, GA3 solution heights 4-6cm；Immersion position is branch base portion.
Described Wpm is xylophyta culture medium, and wherein 10 times of a great number of elements is：Ca(NO3)·4H2O 5.56g/L、
NH4NO3 3.50g/L、K2SO3 9.9g/L、MgSO4·7H2O 3.20g/L、KH2PO4 1.70g/L、CaCl20.725g/L；
100 times of trace element is：MnSO4·4H2O 2.23g/L、ZnSO4·7H20 0.86g/L、H3BO3 0.62g/L、CuSO4·
5H2O 0.025g/L、Na2MoO4·2H2O 0.025g/L；100 times of iron salt is：FeSO4·7H2O 2.78g/L、Na2-EDTA
3.72g/L；Organic 100 times are：Thiamine hydrochloride VB1 0.1g/L, pyridoxine hydrochloride VB6 0.05g/L, nicotinic acid VB5 0.05g/
L, glycine 0.2g/L, inositol 10g/L.
As dove tree lateral bud has long brachyplast phenomenon, when causing indoor cultivation, bud does not extend, undifferentiated, waste of manpower thing
Power, also influence research result.The present invention takes outdoor Hormones pretreatment method, carries out dipping pretreatment by GA3, simply can have
Effect screens length twig problem, moreover it is possible to promote brachyplast bud to sprout elongation to a certain extent, dove tree sprouting of lateral bud is obviously promoted
Effect, also has obvious facilitation to bud elongation.
During existing dove tree the induction of the lateral bud be all take stripping bud after induce sprouting elongation, and GA3 solution pretreatment reduce again
Laboratory stripping bud link, directly carries out inducing clumping bud operation, so as to reduce the consumption of man power and material in experimentation, carries
High work efficiency, has saved production cost.
The culture medium adopted during inoculated and cultured can directly allow the induction of the lateral bud Multiple Buds, and induction time foreshortens to 20 days, averagely
Increment multiplying power is 5.2.In this culture medium be with KT as major influence factors, while reduce agar, cane sugar content to 3.5g, 20g,
And add a small amount of AC, dove tree the induction of the lateral bud Multiple Buds can be obviously promoted.
Due to adopting GA3 pretreatment, breaking dormancy to promote growth；Show through test of many times, KT is to dove tree Growth and Differentiation
There is stronger facilitation；Reduce Nutrient Flow in agar, sucrose concentration energy intensified education base, promote its Growth and Differentiation；Real
Now directly induce.The dove tree inducing clumping bud technology provided using the present invention, method are easy, and induction duration is short, and breeding coefficient is high,
There is certain facilitation to this country-level rare and endangered plant protection.
Description of the drawings
Fig. 1 is photo of the dove tree with hormone immersion treatment outside lateral bud branch room in embodiment 2.
Fig. 2 is growing state when processing rear side bud inoculation 7d in embodiment 2.
Fig. 3 is growing state when processing rear side bud inoculation 15d in embodiment 2.
Fig. 4 is growing state when processing rear side bud inoculation 30d in embodiment 2.
Specific embodiment
With reference to specific embodiment, the present invention is further elucidated with.
Embodiment 1：
1) growth selection stalwartness, no disease and pests harm, character be excellent, lateral bud is formed and dove tree branch during non-lamina is maternal plant,
Add and soaked in GA3 solution, more than 3cm is grown into dove tree lateral bud；
2) clip lateral bud, carries out disinfection, and is then inoculated in Wpm+KT1.2mg L-1+NAA0.3mg·L-1+6-
BA0.5mg·L-1+ agar 3.5g L-1+ sucrose 20g L-1+AC0.2g·L-1Culture medium in；After culture 18-22d, bud point is sprouted
Grow, form Multiple Buds.
Embodiment 2：
1) growth selection stalwartness, no disease and pests harm, character be excellent, lateral bud is formed and dove tree branch during non-lamina is maternal plant,
Soaked in the GA3 solution for adding 100mg/L, soaking solution was changed per 5 days, grow into more than 3cm to dove tree lateral bud；As schemed
Shown in 1；
2) clip lateral bud, during clip, base portion band 1-2mm xylem, carries out disinfection, specifically first flowing water flushing 1h, and 0.4%
Detergent immersion 4-6min, after carry out 75% alcohol-pickled 20s, aseptic water washing 2 times, 0.1%Hgcl in operating board2Immersion 3-
5min, aseptic water washing 4 times, completes disinfecting steps；Then Wpm+KT1.2mg L are inoculated in-1+NAA0.3mg·L-1+6-
BA0.5mg·L-1+ agar 3.5g L-1+ sucrose 20g L-1+AC0.2g·L-1Culture medium in；Culture environment be temperature 25 ±
1 DEG C, humidity 75 ± 2%, illumination 12-14h/d, the constant humiture of light intensity 1800-2200lx culture indoor；After culture 5-7d
Lateral bud base portion starts to expand, and lateral bud continues elongation, and on rear side of culture 10-14d, bastem portion produces 3-7 bud point, bud point after 18-22d
Bud point germination and growth, forms Multiple Buds, and up to 92%, average increment multiple is 5.2 to inducing clumping bud rate, as in Figure 2-4.
Described Wpm is xylophyta culture medium, and wherein 10 times of a great number of elements is：Ca(NO3)·4H2O 5.56g/L、
NH4NO3 3.50g/L、K2SO3 9.9g/L、MgSO4·7H2O 3.20g/L、KH2PO4 1.70g/L、CaCl20.725g/L；
100 times of trace element is：MnSO4·4H2O 2.23g/L、ZnSO4·7H20 0.86g/L、H3BO3 0.62g/L、CuSO4·
5H2O 0.025g/L、Na2MoO4·2H2O 0.025g/L；100 times of iron salt is：FeSO4·7H2O 2.78g/L、Na2-EDTA
3.72g/L；Organic 100 times are：Thiamine hydrochloride VB1 0.1g/L, pyridoxine hydrochloride VB6 0.05g/L, nicotinic acid VB5 0.05g/
L, glycine 0.2g/L, inositol 10g/L.
The optimization of GA3 solution concentrations：
The excellent dove tree of the character such as growth selection stalwartness, no disease and pests harm is maternal plant, intercepts when lateral bud formation and non-lamina
The raw then branch from top to bottom to the 5th lateral bud is experiment material, is a bundle rubber band banding per 10 branches, with clear
Water, tetra- kinds of process of 10mg/LGA3,100mg/L GA3,500mg/L GA3 are soaked, and 5d changes a solution, is united after 10d
Meter germination and growth situation is shown in Table 1.
Table 1
GA3 can be obtained by table one effect is obviously promoted to dove tree sprouting of lateral bud, also have obvious facilitation to bud elongation,
Optimization process is the process of 100mg/LGA3 solution.
The optimization of the operating time of the group culturation rapid propagating technology of Multiple Buds is induced to dove tree：
It is September then to March next year that lateral bud formed, expand time cycle of lamina, in order to find out the optimal acquisition time,
9,10,11,12,1,2, No. 10 difference in March gather dove tree band lateral bud branch as stated above, soaked with 100mg/LGA3 solution
Bubble process, 5d are changed a solution, carry out statistics to lateral bud germination and growth situation and be shown in Table 2 after 10d.
Table 2
Formed to process to outside GA3 rooms used in 7 months of lamina in lateral bud as shown in Table 2 and can effectively promote sprouting of lateral bud
And elongation, when wherein optimal acquisition and process time will be sprouted naturally for March lateral bud, up to 100%, average bud is a length of for germination rate
3.9cm.
The above embodiments are only the preferred technical solution of the present invention, and are not construed as the restriction of the present invention, this
The technical scheme that bright protection domain should be recorded with claim, technical characteristic in the technical scheme that records including claim
Equivalents are protection domain.Equivalent i.e. within this range is improved, also within protection scope of the present invention.
Claims (10)
1. a kind of dove tree efficiently induces the tissue culture and rapid propagation method of Multiple Buds, it is characterised in that comprise the following steps：
1）Growth selection stalwartness, no disease and pests harm, character are excellent, lateral bud is formed and dove tree branch during non-lamina is maternal plant, add
Soaked in GA3 solution, more than 3cm is grown into dove tree lateral bud；
2）Clip lateral bud, carries out disinfection, and is then inoculated in Wpm+KT1.2mg L-1+NAA0.3 mg·L-1+6-BA0.5 mg·
L-1+ agar 3.5g L-1+ sucrose 20g L-1+AC0.2g·L-1Culture medium in；Bud point germination and growth after culture 18-22d, shape
Into Multiple Buds.
2. method according to claim 1, it is characterised in that：The time for dove tree branch being acquired and being processed is for then
September to next year March.
3. method according to claim 1, it is characterised in that：The time for dove tree branch being acquired and being processed is March
Part.
4. method according to claim 1, it is characterised in that：Step 1）Middle dove tree branch is to intercept life then from top to bottom
Branch to the 5th lateral bud.
5. method according to claim 1, it is characterised in that：Step 1）The GA3 solution of 100mg/L is adopted during middle immersion,
A solution was changed per 5 days.
6. method according to claim 1, it is characterised in that：Step 2）The lateral bud of middle clip contains 1-2mm xylem.
7. method according to claim 1, it is characterised in that：Step 2）During middle sterilization, first flowing water rinses 1h, using quality
Concentration is 0.4% detergent immersion 4-6min, then adopts volumetric concentration for 75% alcohol-pickled 20s, aseptic water washing 2 times, then
It is 0.1%Hgcl with mass concentration2Immersion 3-5min, last aseptic water washing 4 times.
8. method according to claim 1, it is characterised in that：The environment of culture be 25 ± 1 DEG C of temperature, humidity 75 ± 2%,
Illumination 12-14h/d, light intensity 1800-2200lx.
9. method according to claim 1, it is characterised in that：Step 1）In immersion process processed in outdoor, GA3
Solution height 4-6cm；Immersion position is branch base portion.
10. method according to claim 1, it is characterised in that：Described Wpm is xylophyta culture medium, wherein in a large number
10 times of element is：Ca（NO3）·4H2O 5.56g/L、NH4NO3 3.50g/L、K2SO3 9.9g/L、MgSO4·7H2O
3.20g/L、KH2PO4 1.70g/L、CaCl20.725g/L；100 times of trace element is：MnSO4·4H2O 2.23g/L、
ZnSO4·7H20 0.86g/L、H3BO3 0.62g/L、CuSO4·5H2O 0.025g/L、Na2MoO4·2H2O 0.025g/
L；100 times of iron salt is：FeSO4·7H2O 2.78g/L、Na2-EDTA 3.72g/L；Organic 100 times are：Thiamine hydrochloride VB1
0.1g/L, pyridoxine hydrochloride VB6 0.05g/L, nicotinic acid VB5 0.05g/L, glycine 0.2g/L, inositol 10g/L.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107324910A (en) * | 2017-08-11 | 2017-11-07 | 仲恺农业工程学院 | A kind of blueberry Foliage nutrient solution and its preparation and spraying method |
| CN107347651A (en) * | 2017-08-30 | 2017-11-17 | 绵阳师范学院 | The cultural method that dove tree hibernaculum quickly breeds |
| CN107494283A (en) * | 2017-10-18 | 2017-12-22 | 贵州省金黔草农业有限公司 | A kind of davidia involucrata var. vilmoriniana tissue culture and rapid propagation method |
| CN111406655A (en) * | 2020-05-26 | 2020-07-14 | 陕西理工大学 | Davidia involucrata seedling and Davidia involucrata tissue culture method |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107324910A (en) * | 2017-08-11 | 2017-11-07 | 仲恺农业工程学院 | A kind of blueberry Foliage nutrient solution and its preparation and spraying method |
| CN107347651A (en) * | 2017-08-30 | 2017-11-17 | 绵阳师范学院 | The cultural method that dove tree hibernaculum quickly breeds |
| CN107494283A (en) * | 2017-10-18 | 2017-12-22 | 贵州省金黔草农业有限公司 | A kind of davidia involucrata var. vilmoriniana tissue culture and rapid propagation method |
| CN111406655A (en) * | 2020-05-26 | 2020-07-14 | 陕西理工大学 | Davidia involucrata seedling and Davidia involucrata tissue culture method |
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| CN106489731B (en) | 2018-11-30 |
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