CN106489731B - 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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- CN106489731B CN106489731B CN201610910782.1A CN201610910782A CN106489731B CN 106489731 B CN106489731 B CN 106489731B CN 201610910782 A CN201610910782 A CN 201610910782A CN 106489731 B CN106489731 B CN 106489731B
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- dove tree
- bud
- lateral bud
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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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- 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 the tissue culture and rapid propagation method that a kind of dove tree efficiently induces Multiple Buds, the specific steps are：1）Dove tree branch when selecting excellent robust growth, no disease and pests harm, character, lateral bud formation and non-lamina is maternal plant, is added in GA3 solution and is impregnated, until dove tree lateral bud grows into 3cm or more；2）Clip lateral bud, carries out disinfection, and is then inoculated in Wpm+KT1.2mgL‑1+NAA0.3mg·L‑1+6‑BA0.5 mg·L‑1+ agar 3.5gL‑1+ sucrose 20gL‑1+AC0.2g·L‑1Culture medium in；Bud point germination and growth after culture 18-22d, forms Multiple Buds.Using dove tree inducing clumping bud technology provided by the invention, method is easy, and induction duration is short, and breeding coefficient is high, has 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 technique
Dove tree is deciduous tree.15~25 meters of height can be grown into, leaf is extensively oval, and there is sawtooth at edge.Dyeing defect is odd beautiful, is
The Relict Plant that the 10000000 years Pre-Cenozoic Tertiary Period left, in quaternary glacier period, the dove tree of most area is gone out in succession
Absolutely, it only survives in some areas of southern china, becomes " living fossil " of plant kingdom's today, be known as " Chinese
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 had been cited as, for the distinctive single platymiscium of China, belongs to Relict Plant and the whole world writes
The ornamental plant of name.
Currently, the reproduction technique of dove tree is mainly based on seeding and seedling raising, but this mode has the following disadvantages：The seedling period
Long, seedling-growing time is 3-10；Low, irregular, the easy variation of emergence rate has certain influence to the merit for saving maternal plant；Together
When need a large amount of human and material resources and land resource, high production cost；In conclusion find emergence fast, the low dove tree of production cost
Tissue-culturing rapid propagation mode pushes industrialization production to have necessity.
Southwestern Normal University's Zou Li Juan exists《Plant research》It delivers《The tissue cultures of Endangered Plant Davidia involucrate Baill and plant are again
It is raw》A kind of quick-breeding method that Multiple Buds are directly induced using dove tree lateral bud is disclosed, but there are explant acquisition times for the method
Disadvantage limited, induction time is long, and its appreciation rate and proliferation times all have a certain upgrade space；Sichuan Agricultural University's summer
What break was delivered《The research of dove tree Initial culture》It is delivered with Hubei ethnic university sieve generation honor《Dove tree sprout Study on tissue culture》In
It all mentions and carries out bud induction by explant of dove tree lateral bud, but can only all simple bud be promoted to sprout, Multiple Buds cannot be induced.
Summary of the invention
It is easy to operate it is an object of the invention to provide the tissue culture and rapid propagation method that a kind of dove tree efficiently induces Multiple Buds, induction
Speed is fast, and growth coefficient is high.
In order to solve the above technical problems, the technical scheme adopted by the invention is that：A kind of dove tree efficiently induces Multiple Buds
Tissue culture and rapid propagation method includes the following steps：
1) dove tree branch when selecting excellent robust growth, no disease and pests harm, character, lateral bud formation and non-lamina is maternal plant,
It is added in GA3 solution and is impregnated, until dove tree lateral bud grows into 3cm or more；
2) clip lateral bud carries out disinfection, and is then inoculated in Wpm+KT1.2mgL-1+NAA0.3mg·L-1+6-
BA0.5mg·L-1+ agar 3.5gL-1+ sucrose 20gL-1+AC0.2g·L-1Culture medium in；Bud point is sprouted after culture 18-22d
It grows, forms Multiple Buds.
The time that dove tree branch is acquired and is handled be current year September to next year March.
Preferably, the time for dove tree branch being acquired and being handled is March.It will sprout naturally for lateral bud at this time
When, germination rate, the long growth of bud is fastly.
Dove tree branch is that interception current year is raw from top to bottom to the branch of the 5th lateral bud in step 1).
Using the GA3 solution of 100mg/L when impregnating in step 1), a solution is changed within every 5 days.
The lateral bud of clip contains 1-2mm xylem in step 2).
In step 2) when disinfection, first flowing water is rinsed 1h and is then adopted using 0.4% detergent immersion 4-6min of mass concentration
20s is impregnated with 75% alcohol of volumetric concentration, aseptic water washing 2 times, then with mass concentration 0.1%Hgcl23-5min is impregnated, 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.
Soaking process in step 1) is handled in outdoor, GA3 solution height 4-6cm；Immersion position is branch base portion.
The Wpm is xylophyta culture medium, is wherein for 10 times of a great number of elements：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 microelement are：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 molysite are：FeSO4·7H2O 2.78g/L,Na2-EDTA
3.72g/L；Organic 100 times are：Thiamine hydrochloride VB1 0.1g/L, puridoxine hydrochloride VB6 0.05g/L, niacin VB5 0.05g/
L, glycine 0.2g/L, inositol 10g/L.
Since dove tree lateral bud is there are long brachyplast phenomenon, bud when indoor culture is caused not extend, undifferentiated, waste of manpower object
Power also influences result of study.The present invention takes outdoor Hormones pretreatment method, carries out dipping pretreatment by GA3, can simply have
Effect screens length twig problem, moreover it is possible to promote brachyplast bud to sprout elongation to a certain extent, be obviously promoted to the sprouting of dove tree lateral bud
Effect also has apparent facilitation to bud elongation.
It is all to induce sprouting to extend after taking stripping bud again, and GA3 solution pre-processes, and can reduce when existing dove tree the induction of the lateral bud
Laboratory stripping bud link, directly progress inducing clumping bud process mention to reduce the consumption of man power and material in experimentation
High working efficiency, has saved production cost.
The culture medium used when inoculated and cultured can directly allow the induction of the lateral bud Multiple Buds, and induction time foreshortens to 20 days, average
Multiplying power of rising in value is 5.2.It is while to reduce agar, cane sugar content to 3.5g, 20g using KT as major influence factors in this culture medium,
And a small amount of AC is added, dove tree the induction of the lateral bud Multiple Buds can be obviously promoted.
Due to being pre-processed using GA3, breaking dormancy promotes growth；Show KT to dove tree Growth and Differentiation by test of many times
With stronger facilitation；Nutrient Flow in reduction agar, sucrose concentration energy intensified education base, promotes its Growth and Differentiation；It is real
Now directly induce.Using dove tree inducing clumping bud technology provided by the invention, method is easy, and induction duration is short, and breeding coefficient is high,
There is certain facilitation to this country-level rare and endangered plant protection.
Detailed description of the invention
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 is inoculated with 7d in embodiment 2.
Fig. 3 is growing state when processing rear side bud is inoculated with 15d in embodiment 2.
Fig. 4 is growing state when processing rear side bud is inoculated with 30d in embodiment 2.
Specific embodiment
Combined with specific embodiments below, the present invention is furture elucidated.
Embodiment 1：
1) dove tree branch when selecting excellent robust growth, no disease and pests harm, character, lateral bud formation and non-lamina is maternal plant,
It is added in GA3 solution and is impregnated, until dove tree lateral bud grows into 3cm or more；
2) clip lateral bud carries out disinfection, and is then inoculated in Wpm+KT1.2mgL-1+NAA0.3mg·L-1+6-
BA0.5mg·L-1+ agar 3.5gL-1+ sucrose 20gL-1+AC0.2g·L-1Culture medium in；Bud point is sprouted after culture 18-22d
It grows, forms Multiple Buds.
Embodiment 2：
1) dove tree branch when selecting excellent robust growth, no disease and pests harm, character, lateral bud formation and non-lamina is maternal plant,
It is added in the GA3 solution of 100mg/L and is impregnated, every 5 days replacement soaking solutions, until dove tree lateral bud grows into 3cm or more；Such as figure
Shown in 1；
2) clip lateral bud, base portion band 1-2mm xylem, carries out disinfection when clip, and specifically first flowing water rinses 1h, and 0.4%
Detergent immersion 4-6min, after station carry out 75% alcohol impregnate 20s, aseptic water washing 2 times, 0.1%Hgcl2Impregnate 3-
5min aseptic water washing 4 times, completes disinfecting steps；Then it is inoculated in Wpm+KT1.2mgL-1+NAA0.3mg·L-1+6-
BA0.5mg·L-1+ agar 3.5gL-1+ sucrose 20gL-1+AC0.2g·L-1Culture medium in；Culture environment be temperature 25 ±
1 DEG C, humidity 75 ± 2%, the constant temperature and humidity of illumination 12-14h/d, light intensity 1800-2200lx culturing room in；After cultivating 5-7d
Lateral bud base portion starts to expand, and lateral bud continues to extend, and bastem portion generates 3-7 bud point, bud point after 18-22d on rear side of culture 10-14d
Bud point germination and growth forms Multiple Buds, and inducing clumping bud rate is up to 92%, and the multiple that averagely rises in value is 5.2, as in Figure 2-4.
The Wpm is xylophyta culture medium, is wherein for 10 times of a great number of elements：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 microelement are：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 molysite are：FeSO4·7H2O 2.78g/L,Na2-EDTA
3.72g/L；Organic 100 times are：Thiamine hydrochloride VB1 0.1g/L, puridoxine hydrochloride VB6 0.05g/L, niacin VB5 0.05g/
L, glycine 0.2g/L, inositol 10g/L.
The optimization of GA3 solution concentration：
The dove tree for selecting the characters such as robust growth, no disease and pests harm excellent is maternal plant, is intercepted when lateral bud formation and non-lamina
Current year raw is from top to bottom experimental material to the branch of the 5th lateral bud, and every 10 branches are to use rubber band banding a bundle, with clear
Tetra- kinds of water, 10mg/LGA3,100mg/L GA3,500mg/L GA3 processing are impregnated, and 5d changes a solution, unites 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 the sprouting of dove tree lateral bud, also have apparent facilitation to bud elongation,
Optimization process is the processing of 100mg/LGA3 solution.
Optimization to the operating time of the group culturation rapid propagating technology of dove tree induction Multiple Buds：
It is current year September to March next year that lateral bud, which forms, expands the time cycle of lamina, in order to find out the optimal acquisition time,
9,10,11,12,1,2, No. 10 difference in March acquire dove tree band lateral bud branch according to the above method, soaked with 100mg/LGA3 solution
Bubble processing, 5d change a solution, carry out statistics to lateral bud germination and growth situation after 10d and be shown in Table 2.
Table 2
Effectively lateral bud can be promoted to sprout using processing outside the room GA3 in 7 middle of the month that lateral bud is formed to lamina as shown in Table 2
And elongation, when wherein optimal acquisition and processing time are that lateral bud in March will sprout naturally, for germination rate up to 100%, average bud is a length of
3.9cm。
The above embodiments are only the preferred technical solution of the present invention, and are not construed as limitation of the invention, this hair
Bright protection scope should be with the technical solution of claim record, technical characteristic in the technical solution recorded including claim
Equivalents are protection scope.Equivalent replacement i.e. within this range is improved, also within protection scope of the present invention.
Claims (8)
1. the tissue culture and rapid propagation method that a kind of dove tree efficiently induces Multiple Buds, which is characterized in that include the following steps：
1）Dove tree branch when selecting excellent robust growth, no disease and pests harm, character, lateral bud formation and non-lamina is maternal plant, is added
GA3It is impregnated in solution, until dove tree lateral bud grows into 3cm or more；Soaking process is handled in outdoor, GA3Solution height 4-
6cm；Immersion position is branch base portion；
2）Clip lateral bud, carries out disinfection, and is then inoculated in Wpm+KT1.2mgL-1+NAA0.3 mg·L-1+6-BA0.5 mg·
L-1+ agar 3.5gL-1+ sucrose 20gL-1+AC0.2g·L-1Culture medium in；Bud point germination and growth after culture 18-22d, shape
At Multiple Buds；
The Wpm is xylophyta culture medium, is wherein for 10 times of a great number of elements：Ca（NO3）2·4H2O 5.56g/L、NH4NO3
3.50g/L、K2SO3 9.9g/L、MgSO4·7H2O 3.20g/L、KH2PO4 1.70g/L、CaCl20.725g/L；Microelement
100 times are：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 molysite are：FeSO4·7H2O 2.78g/L、Na2-EDTA 3.72g/
L；Organic 100 times are：Thiamine hydrochloride VB1 0.1g/L, puridoxine hydrochloride VB6 0.05g/L, niacin VB5 0.05g/L, sweet ammonia
Sour 0.2g/L, inositol 10g/L.
2. according to the method described in claim 1, it is characterized in that：The time that dove tree branch is acquired and is handled is current year
September to next year March.
3. according to the method described in claim 1, it is characterized in that：The time that dove tree branch is acquired and is handled is March
Part.
4. according to the method described in claim 1, it is characterized in that：Step 1）Middle dove tree branch is that interception current year is raw from top to bottom
To the branch of the 5th lateral bud.
5. according to the method described in claim 1, it is characterized in that：Step 1）Using the GA of 100mg/L when middle immersion3Solution,
Change a solution within every 5 days.
6. according to the method described in claim 1, it is characterized in that：Step 2）The lateral bud of middle clip contains 1-2mm xylem.
7. according to the method described in claim 1, it is characterized in that：Step 2）When middle disinfection, first flowing water rinses 1h, using quality
Concentration is 0.4% detergent immersion 4-6min, then uses volumetric concentration for 75% alcohol immersion 20s, aseptic water washing 2 times, then
It is 0.1%HgCl with mass concentration2Impregnate 3-5min, last aseptic water washing 4 times.
8. according to the method described in claim 1, it is characterized in that：The environment of culture be 25 ± 1 DEG C of temperature, humidity 75 ± 2%,
Illumination 12-14h/d, light intensity 1800-2200lx.
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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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