CN107683768A - A kind of acclimatization and transplantses method of the syringa reticulata var mandshurica tissue-cultured seedling of callus induction - Google Patents
A kind of acclimatization and transplantses method of the syringa reticulata var mandshurica tissue-cultured seedling of callus induction 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
- A01H4/008—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
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Abstract
The invention provides a kind of acclimatization and transplantses method of the syringa reticulata var mandshurica tissue-cultured seedling of callus induction, belong to plant tissue culture technical field.The acclimatization and transplantses method of the syringa reticulata var mandshurica tissue-cultured seedling of callus induction provided by the invention includes：Take the syringa reticulata var mandshurica tissue-cultured seedling of callus induction to carry out 5~10d of hardening, obtain seedling after hardening, the temperature of the hardening is 15~18 DEG C, and intensity of illumination is 800~1000lx, and daily 5~8h of illumination, hardening humidity is 70~80%；By small transplantation of seedlings after the obtained hardening into transplanting medium, nutrient solution is added into the matrix of seedling root daily.By the acclimatization and transplantses process of the present invention, the transplanting survival rate of the syringa reticulata var mandshurica tissue-cultured seedling of callus induction is up to more than 91%, acclimatization and transplantses method provided by the invention is simple to operate, and cost of labor is low, and survival rate is low after solving the problems, such as the tissue culture transplantation of seedlings of syringa reticulata var mandshurica callus induction.
Description
Technical field
The invention belongs to a kind of biological tissue culture technology field, a kind of syringa reticulata var mandshurica group of callus induction is more particularly to
Train the acclimatization and transplantses method of seedling.
Background technology
Syringa reticulata var mandshurica Syringareticulata (Blume), also known as Syringa amurensis, white lilac, fallen leaves for Oleaceae cloves tree
Dungarunga.About 35 kinds of the cloves tree whole world, is distributed mainly on the Temperate Region in China in Asia and Europe, and China produces more than 25 kinds.Sudden and violent horse
Cloves, because its is with luxuriant foliage and spreading branches in leafy profusion, inflorescence is very large, giving off a strong fragrance attacks people, is using more universal in northern China gardens as a kind of ornamental plant
One of seeds, in addition, the solid densification of its material, structure are homogeneous；Essential oil content enriches, and has extensive bioactivity；Sudden and violent horse
The landscape value and economic value of cloves are increasingly subject to the attention of people.Syringa reticulata var mandshurica mainly uses conventional propagation method, with kind
Son breeding and the asexual process such as cuttage, grafting.But the seed used in seminal propagation needs complicated pregermination procedure, and emerges not
Uniformly, primary growth is slow and solid the features such as being influenceed by biennial bearing so that the more sight flower type of this veriety of cloves, due to
Itself it is mostly heterozygosis genotype, therefore seminal propagation can not keep the typical character of its kind.
At present, there is certain research to syringa reticulata var mandshurica cultured in vitro, it is true using bud propagation and two approach of callus
The syringa reticulata var mandshurica Regeneration of a set of more system is found, but tissue-cultured seedling is in the tissue culture ambient condition of sterile moistening
Under, root system generates adaptability to the environment that moisture around is sufficient, nutrient is abundant, and secondly, test tube seedling is smaller, to poor environment
Buffer capacity is extremely limited, and resistance is poor, is directly transplanted in matrix to be put into natural environment and is very easy to cause water shortage to wither,
Reduce survival rate；In addition, for syringa reticulata var mandshurica, the tissue-cultured seedling that the tissue-cultured seedling of callus induction is induced than general stem section is more
It is weak, the transplanting death rate is exacerbated, makes survival rate lower.
The content of the invention
In view of this, it is an object of the invention to provide a kind of shifting of the hardening of the syringa reticulata var mandshurica tissue-cultured seedling of callus induction
Cultivating method, the transplanting death rate is reduced, improves survival rate.
In order to realize foregoing invention purpose, the present invention provides following technical scheme：
The invention provides a kind of acclimatization and transplantses method of the syringa reticulata var mandshurica tissue-cultured seedling of callus induction, including：
1) take the syringa reticulata var mandshurica tissue-cultured seedling of callus induction to carry out 5~10d of hardening, obtain seedling after hardening, the hardening
Temperature be 15~18 DEG C, intensity of illumination be 800~1000lx, daily 5~8h of illumination, and hardening humidity is 70~80%；
2) small transplantation of seedlings is into transplanting medium after the hardening for obtaining step 1), daily into the matrix of seedling root
Add nutrient solution；
The transplanting medium includes a kind of and mixture of turfy soil in river sand and vermiculite；
The nutrient solution includes 2mg/L's using 1/3MS fluid nutrient mediums and 1/3WPM fluid nutrient mediums as minimal medium
IBA and 1mg/L NAA.
Preferably, the tissue-cultured seedling of the callus induction, height of seedling are not less than 3cm, and radical is no less than 4.
Preferably, step 2) the nutrient solution addition is in the surface of the transplanting medium, the positional distance institute of the addition
State 0.8~1.5cm of seedling.
Preferably, the step 2) nutrient solution adds 13~16d altogether, and the amount added daily is 15~25ml.
Preferably, the breeding method of the tissue-cultured seedling, comprises the following steps：
A) take syringa reticulata var mandshurica development hypocotyl be seeded on callus inducing medium carry out Fiber differentiation 40~
60d, obtain callus；
B) the obtained callus of the step a) is inoculated on adventitious bud induction culture base, cultivates 40~55d, obtain more
The syringa reticulata var mandshurica tissue-cultured seedling of injured tissue induction.
Preferably, the culture medium of the step a) Fiber differentiations is using MS solid mediums as minimal medium, in addition to
5mg/L BA and 0.5mg/L IBA.
Preferably, the condition of the step a) Fiber differentiations is：20~24 DEG C, intensity of illumination 2000lx of temperature, during illumination
Between daily 10~12h.
Preferably, the acquisition methods of the development hypocotyl comprise the following steps：Seed after sterilization is inoculated in into MS to consolidate
After carrying out cultured in vitro 10d on body culture medium, cut to obtain development hypocotyl.
Preferably, to clean up the seed, clear water rinses 1~2h, is then with volumn concentration for the sterilization
75% ethanol immersion 30s, aseptic water washing 3 times, after air-drying again with the mercuric chloride of weight/mass percentage composition 0.1% sterilization 5~
10min, aseptic water washing 3~5 times.
Preferably, the syringa reticulata var mandshurica tissue-cultured seedling of the callus induction moves before hardening, in addition to by the tissue-cultured seedling
Plant and strong seedling culture is carried out on strong seedling culture base, the time of the strong seedling culture is 7~12d, and the strong seedling culture base is with 1/6MS
Solid medium and 1/6WPM solid mediums are minimal medium, and pH value is 5.8~6.5.
The invention provides a kind of acclimatization and transplantses method of the syringa reticulata var mandshurica tissue-cultured seedling of callus induction.The present invention provides
Acclimatization and transplantses method will obtain tissue-cultured seedling in sterile moist environment, by hardening so that tissue-cultured seedling has one to external environment condition
Fixed buffer capacity, improve resistance；During hardening, nutrient solution is added to seedling root, one kind is formed and is advantageous to plant growth
External nutrition environment, ensure the root absorption of test tube seedling enough nutrient and moisture, not only saved cost, also reduce transplanting
The death rate, survival rate is improved to more than 91%.
Embodiment
The invention provides a kind of acclimatization and transplantses method of the syringa reticulata var mandshurica tissue-cultured seedling of callus induction, including：
1) take the syringa reticulata var mandshurica tissue-cultured seedling of callus induction to carry out 5~10d of hardening, obtain seedling after hardening, the hardening
Temperature be 15~18 DEG C, intensity of illumination be 800~1000lx, daily 5~8h of illumination, and hardening humidity is 70~80%；
2) small transplantation of seedlings is into transplanting medium after the hardening for obtaining step 1), daily into the matrix of seedling root
Add nutrient solution；
The transplanting medium includes a kind of and mixture of turfy soil in river sand and vermiculite；
The nutrient solution includes 2mg/L's using 1/3MS fluid nutrient mediums and 1/3WPM fluid nutrient mediums as minimal medium
IBA and 1mg/L NAA.
The present invention preferably carries out the cultivation of tissue-cultured seedling before acclimatization and transplantses are carried out.In the present invention, the syringa reticulata var mandshurica is cured
The breeding method of the tissue-cultured seedling of injured tissue induction, comprises the following steps：
A) take syringa reticulata var mandshurica development hypocotyl be seeded on callus inducing medium carry out Fiber differentiation 40~
60d, obtain callus；
B) the obtained callus of the step a) is inoculated on adventitious bud induction culture base, cultivates 40~55d, obtain more
The syringa reticulata var mandshurica tissue-cultured seedling of injured tissue induction.
The present invention takes the development hypocotyl of syringa reticulata var mandshurica to be seeded to progress Fiber differentiation 40 on callus inducing medium
~60d, obtains callus.In the present invention, the time of the Fiber differentiation is preferably 50d.In the present invention, under the development
The acquisition methods of plumular axis preferably include：Syringa reticulata var mandshurica seed after sterilization is inoculated on MS solid mediums and carries out cultured in vitro
Cut to obtain development hypocotyl after 10d.The present invention does not have special restriction to the source of the syringa reticulata var mandshurica seed, using ability
Syringa reticulata var mandshurica seed conventional commercial product known to field technique personnel.
In the present invention, the sterilization method of syringa reticulata var mandshurica seed is preferably：Cleaning up the seed, clear water rinses 1~
2h, 30s then is soaked with the ethanol that volumn concentration is 75%, aseptic water washing 3 times, weight/mass percentage composition is used again after air-drying
0.1% mercuric chloride sterilizes 5~10min, aseptic water washing 3~5 times.
After the seed being sterilized, the present invention cuts the seed of sterilization, cuts hypocotyl, after obtaining hypocotyl, this hair
It is bright that hypocotyl is preferably placed in progress hypocotyl cultured in vitro in cultured in vitro base before induction of callus is carried out,
In the present invention, the culture medium of the cultured in vitro is preferably MS solid mediums, and the condition of the hypocotyl cultured in vitro is preferred
For：20~24 DEG C, intensity of illumination 2000lx of cultivation temperature, the daily 12h of light application time；The time of the cultured in vitro is preferred
10d；After the hypocotyl developed.The hypocotyl of development is preferably cut into 5~7mm section by the present invention, and access callus lures
Lead and induction of callus is carried out on culture medium, the callus inducing medium is basic preferably using MS solid mediums
Culture medium, include 5mg/L BA and 0.5mg/L IBA；The condition of the Fiber differentiation is preferably：20~24 DEG C of temperature, light
According to intensity 2000lx, the daily 10~12h of light application time, light application time is preferably daily 12h.
After obtaining callus, obtained callus is inoculated on adventitious bud induction culture base by the present invention, culture 40
~55d, obtain the syringa reticulata var mandshurica tissue-cultured seedling of callus induction.In the present invention, the incubation time is preferably 46~52d, more
Preferably 50d.In the present invention, the callus, which is preferably chosen, grows fine and close, emerald green, loose callus.In this hair
In bright, the adventitious bud induction culture base is preferably using 1/2MS solid mediums as minimal medium, and 2,4- comprising 0.1mg/L
D, 3mg/L BA and 0.5mg/L NAA；The condition of the culture is：20~24 DEG C, intensity of illumination 2000lx of temperature, during illumination
Between daily 12h.
After obtaining the syringa reticulata var mandshurica tissue-cultured seedling of callus induction, in the syringa reticulata var mandshurica tissue-cultured seedling of the callus induction
Before hardening, tissue-cultured seedling is preferably transplanted on strong seedling culture base and carries out strong seedling culture progress strong seedling culture by the present invention.In this hair
In bright, the time of the strong seedling culture is preferably 7~12d, more preferably 10d.In the present invention, the strong seedling culture base is with 1/
6MS solid mediums and 1/6WPM solid mediums are minimal medium, in the present invention, the 1/6MS solid mediums and
The volume ratio of 1/6WPM solid mediums is preferably (1~2):(1~3), more preferably 1:1；The pH value of the strong seedling culture base
Preferably 5.8~6.5, more preferably 5.8.In the present invention, sugar and hormone are not added in the strong seedling culture base, inorganic
The ion concentration of salt reduces, and can promote rooting of vitro seedling, the survival rate of seedling after increase transplanting.
The present invention takes the syringa reticulata var mandshurica tissue-cultured seedling of callus induction to carry out 5~10d of hardening, obtains seedling after hardening, described
The temperature of hardening is 15~18 DEG C, and intensity of illumination is 800~1000lx, and daily 5~8h of illumination, hardening humidity is 70~80%.
In the present invention, the temperature of the hardening is preferably 16 DEG C；In the present invention, the intensity of illumination is preferably 900lx, per the daylight
It is preferably 6h according to the time；In the present invention, the hardening humidity is preferably 75%.In the present invention, the callus induction
Tissue-cultured seedling, height of seedling is preferably not less than 3cm, and radical is preferably no less than 4, and the tissue-cultured seedling preferably chooses well-grown tissue culture
Seedling carries out hardening.The present invention preferably carries out hardening in Sterile culture room.
Obtain small after seedling after hardening, the present invention by small transplantation of seedlings after the hardening into transplanting medium, daily to seedling root
Nutrient solution is added in the matrix in portion；The nutrient solution is using 1/3MS fluid nutrient mediums and 1/3WPM fluid nutrient mediums as basic culture
The volume ratio of base, described/3MS fluid nutrient mediums and 1/3WPM fluid nutrient mediums is preferably (1~3)：(1~2), more preferably
1:1；2mg/L IBA and 1mg/L NAA are preferably comprised in the nutrient solution.In the present invention, the transplanting is preferably transplanted to
In diameter 12cm nutritive cube, seedling after one transplanting of preferable transplanting in each nutritive cube.In the present invention, the shifting
Planting matrix includes a kind of and mixture of turfy soil in river sand and vermiculite, i.e., described transplanting medium includes river sand and turfy soil
Mixture, or the mixture of vermiculite and turfy soil, when the transplanting medium is the mixture of river sand and turfy soil, the river sand
Volume ratio with turfy soil is preferably (1~3)：(3~8), more preferably (2~3)：(4~5), most preferably 2:5；When described
When transplanting medium is the mixture of vermiculite and turfy soil, the volume ratio of the vermiculite and turfy soil is preferably (1~3)：(3~8),
More preferably (2~3)：(4~5), most preferably 2:5.In the present invention, river sand, vermiculite, turfy soil price are relatively low, purchaser
Just, and turfy soil good water-retaining property, but if matrix is all turfy soil, easily cause rotten of tissue-cultured seedling, when add river sand or
After vermiculite, the water penetration of turfy soil is improved, is advantageous to improve the transplanting survival rate of tissue-cultured seedling.In the present invention, the transplanting
Matrix, preferably through 200 DEG C of sterilization 30min of high temperature, then naturally cools to room temperature before use；Matrix of the present invention makes
Preferably matrix is irrigated with water with preceding, specifically reaches matrix and drips；
Heretofore described nutrient solution addition is in the surface of the transplanting medium, seedling described in the positional distance of the addition
0.8~1.5cm.In the present invention, the addition manner of the nutrient solution is preferably to inject, and the position of the injection is transplanting medium
Surface, preferably apart from the 0.8~1.5cm of seedling, more preferably in the matrix at the seedling 1cm, the note
The amount penetrated is preferably every seedling 15~25ml/ times, more preferably 20ml/ times；In the present invention, described inject is preferably
Every morning injects, preferred every morning 7:30 injections；In the present invention, the injection is preferably that continuity is injected, institute
The time for stating injection is preferably 13~16d, more preferably 15d.The mode of injection of the present invention can be formed seedling root
One relative high nutrition, the environment of high humidity, are advantageous to plant growth, and 20ml injection rate is unlikely to form too high high humidity environment
So as to cause rotten, the nutrient solution is locally being used, it is ensured that the root absorption of test tube seedling enough nutrient and moisture, greatly
The death rate is reduced greatly, makes survival rate to more than 91%.
A kind of hardening of the syringa reticulata var mandshurica tissue-cultured seedling of callus induction provided by the invention is moved with reference to embodiment
Cultivating method is described in detail, but they can not be interpreted as limiting the scope of the present invention.
Embodiment 1
(1) seed of the ripe syringa reticulata var mandshurica of selection, is cleaned up using washing powder, then rinses 1h with clear water again；Put
In using the ethanol that volumn concentration is 75% to soak 30s on superclean bench, with aseptic water washing 3 times, aseptic filter paper is placed on
It is upper to air-dry, then with weight/mass percentage composition be that 0.1% mercuric chloride sterilizes 8min, then with aseptic water washing 3 times, seed after must sterilizing；
(2) seed sterilized is cut, hypocotyl is accessed in MS culture mediums and cultivated, condition of culture is：Cultivation temperature
For 22 DEG C, intensity of illumination 2000lx, the daily 12h of light application time, cultivate 10 days, hypocotyl after must developing；
(3) take 5mm hypocotyl to be seeded in using MS solid mediums as minimal medium, include 5mg/LBA, 0.5mg/
It is 20 DEG C, intensity of illumination 2000lx, daily illumination 12h in temperature on LIBA callus inducing medium, culture 50
My god, obtain callus；
(4) it is inoculated into fine and close, emerald green, loose callus is grown using 1/2MS culture mediums as minimal medium, comprising
Have the 2 of 0.1mg/L, be 20 DEG C in temperature on 4-D, 3mg/L BA and 0.5mg/L NAA adventitious bud induction culture base, light
It is 2000lx according to intensity, Fiber differentiation 50d under the conditions of daily illumination 12h, obtains the syringa reticulata var mandshurica tissue-cultured seedling of callus induction；
(5) tissue-cultured seedling is transferred to using 1/6MS solid mediums and 1/6WPM solid mediums as minimal medium again
It is 20 DEG C in temperature in mixed culture medium, intensity of illumination 2000lx, daily illumination 12h, strong seedling culture 10d；
(6) in sterile, 15 DEG C of temperature, intensity of illumination 800lx, daily illumination 5h, under the environment of indoor humidity 70%, by seedling
It is transplanted into river sand and turfy soil volume ratio 2:In 5 transplanting culture medium, hardening 5d is transplanted, obtains seedling after hardening, during hardening,
Daily to the stromal surface injection apart from seedling 1cm using 1/3MS fluid nutrient mediums and 1/3WPM fluid nutrient mediums as basic culture
Base, stop injection after the NAA of IBA and 1mg/L comprising 2mg/L nutrient solution 20ml, 15d, transplanting survival rate is up to 91%.
Embodiment 2
(1) seed of the ripe syringa reticulata var mandshurica of selection, is cleaned up using washing powder, then rinses 2h with clear water again；Put
In using the ethanol that volumn concentration is 75% to soak 30s on superclean bench, with aseptic water washing 3 times, aseptic filter paper is placed on
It is upper to air-dry, then with weight/mass percentage composition be that 0.1% mercuric chloride sterilizes 8min, then with aseptic water washing 3 times, seed after must sterilizing；
(2) seed sterilized is cut, hypocotyl is accessed in MS culture mediums and cultivated, condition of culture is：Cultivation temperature
For 20 DEG C, intensity of illumination 2000lx, the daily 12h of light application time, cultivate 10 days, hypocotyl after must developing；
(3) take 6mm hypocotyl to be seeded in using MS solid mediums as minimal medium, include 5mg/LBA, 0.5mg/
It is 20 DEG C, intensity of illumination 2000lx, daily illumination 12h in temperature on LIBA callus inducing medium, culture 40
My god, obtain callus；
(4) it is inoculated into fine and close, emerald green, loose callus is grown using 1/2MS culture mediums as minimal medium, comprising
Have the 2 of 0.1mg/L, be 22 DEG C in temperature on 4-D, 3mg/L BA and 0.5mg/L NAA adventitious bud induction culture base, light
It is 2000lx according to intensity, Fiber differentiation 40d under the conditions of daily illumination 12h, obtains the syringa reticulata var mandshurica tissue-cultured seedling of callus induction；
(5) tissue-cultured seedling is transferred to using 1/6MS solid mediums and 1/6WPM solid mediums as minimal medium again
It is 22 DEG C in temperature in mixed culture medium, intensity of illumination 2000lx, daily illumination 12h, strong seedling culture 7d；
(6) in sterile, 16 DEG C of temperature, intensity of illumination 900lx, daily illumination 5h, under the environment of indoor humidity 70%, by seedling
It is transplanted into river sand and turfy soil volume ratio 3:In 7 transplanting culture medium, hardening 7d is transplanted, obtains seedling after hardening, during hardening,
Daily to the stromal surface injection apart from seedling 1cm using 1/3MS fluid nutrient mediums and 1/3WPM fluid nutrient mediums as basic culture
Base, stop injection after the NAA of IBA and 1mg/L comprising 2mg/L nutrient solution 15ml, 13d, transplanting survival rate is up to 93%.
Embodiment 3
(1) seed of the ripe syringa reticulata var mandshurica of selection, is cleaned up using washing powder, then rinses 1.5h with clear water again；
It is placed on superclean bench and soaks 30s with the ethanol that volumn concentration is 75%, with aseptic water washing 5 times, is placed on sterile filter
Air-dried on paper, then with weight/mass percentage composition be that 0.1% mercuric chloride sterilizes 8min, then with aseptic water washing 3 times, planted after must sterilizing
Son；
(2) seed sterilized is cut, hypocotyl is accessed in MS culture mediums and cultivated, condition of culture is：Cultivation temperature
For 24 DEG C, intensity of illumination 2000lx, the daily 12h of light application time, cultivate 10 days, hypocotyl after must developing；
(3) take 7mm hypocotyl to be seeded in using MS solid mediums as minimal medium, include 5mg/LBA, 0.5mg/
It is 20 DEG C, intensity of illumination 2000lx, daily illumination 12h in temperature on LIBA callus inducing medium, culture 60
My god, obtain callus；
(4) it is inoculated into fine and close, emerald green, loose callus is grown using 1/2MS culture mediums as minimal medium, comprising
Have the 2 of 0.1mg/L, be 24 DEG C in temperature on 4-D, 3mg/L BA and 0.5mg/L NAA adventitious bud induction culture base, light
It is 2000lx according to intensity, Fiber differentiation 55d under the conditions of daily illumination 12h, obtains the syringa reticulata var mandshurica tissue-cultured seedling of callus induction；
(5) tissue-cultured seedling is transferred to using 1/6MS solid mediums and 1/6WPM solid mediums as minimal medium again
It is 20 DEG C in temperature in mixed culture medium, intensity of illumination 2000lx, daily illumination 12h, strong seedling culture 12d；
(6) in sterile, 15 DEG C of temperature, intensity of illumination 1000lx, daily illumination 8h, under the environment of indoor humidity 80%, by children
Transplantation of seedlings enters river sand and turfy soil volume ratio 2:In 5 transplanting culture medium, hardening 10d is transplanted, obtains seedling after hardening, hardening process
In, daily to the stromal surface injection apart from seedling 1cm using 1/3MS fluid nutrient mediums and 1/3WPM fluid nutrient mediums as basic training
Base is supported, stops injection after the NAA of IBA and 1mg/L comprising 2mg/L nutrient solution 25ml, 16d, transplanting survival rate is reachable
92.6%.
Reference examples
Selection carries out hardening with the identical tissue-cultured seedling of embodiment 1~3, and the sealed membrane of tissue culture bottle is removed, seedling is taken out
Clean up root system；Avoiding direct sunlight, 16 ± 2 DEG C of temperature, in the environment of humidity 80%, be transplanted into turfy soil matrix,
After 15 days, plant strain growth gesture is weaker, leaf some yellowing, beats listless, overall growing state is weaker, and the survival rate for recording seedling is
45.6%.
As seen from the above embodiment, moved using the hardening of the syringa reticulata var mandshurica tissue-cultured seedling of callus induction provided by the invention
Cultivating method, it is significantly higher than comparative example, transplanting survival rate average out to more than 91% in survival rate.
Described above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, under the premise without departing from the principles of the invention, some improvements and modifications can also be made, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (10)
1. a kind of acclimatization and transplantses method of the syringa reticulata var mandshurica tissue-cultured seedling of callus induction, including：
1) take the syringa reticulata var mandshurica tissue-cultured seedling of callus induction to carry out 5~10d of hardening, obtain seedling after hardening, the temperature of the hardening
Spend for 15~18 DEG C, intensity of illumination is 800~1000lx, and daily 5~8h of illumination, hardening humidity is 70~80%；
2) small transplantation of seedlings into transplanting medium, is added into the matrix of seedling root daily after the hardening for obtaining step 1)
Nutrient solution；
The transplanting medium includes a kind of and mixture of turfy soil in river sand and vermiculite；
The nutrient solution is using 1/3MS fluid nutrient mediums and 1/3WPM fluid nutrient mediums as minimal medium, the IBA comprising 2mg/L
With 1mg/L NAA.
2. acclimatization and transplantses method according to claim 1, it is characterised in that the tissue-cultured seedling of the callus induction, seedling
Height is not less than 3cm, and radical is no less than 4.
3. acclimatization and transplantses method according to claim 1, it is characterised in that step 2) the nutrient solution addition is in the shifting
Plant the surface of matrix, 0.8~1.5cm of seedling described in the positional distance of the addition.
4. acclimatization and transplantses method according to claim 1, it is characterised in that the step 2) nutrient solution adds 13 altogether~
16d, the amount added daily are 15~25ml.
5. acclimatization and transplantses method according to claim 1, it is characterised in that the breeding method of the tissue-cultured seedling, including with
Lower step：
A) take the development hypocotyl of syringa reticulata var mandshurica to be seeded to progress 40~60d of Fiber differentiation on callus inducing medium, obtain
Callus；
B) the obtained callus of the step a) is inoculated on adventitious bud induction culture base, cultivates 40~55d, obtain callus group
Knit the syringa reticulata var mandshurica tissue-cultured seedling of induction.
6. acclimatization and transplantses method according to claim 5, it is characterised in that the culture medium of the step a) Fiber differentiations with
MS solid mediums are minimal medium, include 5mg/L BA and 0.5mg/L IBA.
7. acclimatization and transplantses method according to claim 5, it is characterised in that the condition of the step a) Fiber differentiations is：
20~24 DEG C, intensity of illumination 2000lx of temperature, the daily 10~12h of light application time.
8. acclimatization and transplantses method according to claim 5, it is characterised in that the acquisition side of step a) the development hypocotyls
Method comprises the following steps：Seed after sterilization is inoculated in after carrying out cultured in vitro 10d on MS solid mediums, cuts and sent out
Educate hypocotyl.
9. acclimatization and transplantses method according to claim 8, it is characterised in that the sterilization is to clean up the seed,
Clear water rinses 1~2h, then soaks 30s with the ethanol that volumn concentration is 75%, aseptic water washing 3 times, is used again after air-drying
The mercuric chloride of weight/mass percentage composition 0.1% sterilizes 5~10min, aseptic water washing 3~5 times.
10. acclimatization and transplantses method according to claim 1 or 5, it is characterised in that the sudden and violent Martin of the callus induction
Fragrant tissue-cultured seedling before hardening, in addition to：The tissue-cultured seedling is transplanted on strong seedling culture base and carries out strong seedling culture, the strong sprout training
The foster time is 7~12d, and the strong seedling culture base is using 1/6MS solid mediums and 1/6WPM solid mediums as basic culture
Base, pH value are 5.8~6.5.
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| CN108668901A (en) * | 2018-06-20 | 2018-10-19 | 红艳 | Numerous method is expanded in the cloves stem apex regeneration of anaesthetic Helan Mountain |
| CN110771510A (en) * | 2019-11-26 | 2020-02-11 | 大连大学 | Method for preparing artificial clove seeds |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108668901A (en) * | 2018-06-20 | 2018-10-19 | 红艳 | Numerous method is expanded in the cloves stem apex regeneration of anaesthetic Helan Mountain |
| CN110771510A (en) * | 2019-11-26 | 2020-02-11 | 大连大学 | Method for preparing artificial clove seeds |
| CN110771510B (en) * | 2019-11-26 | 2022-08-05 | 大连大学 | Method for preparing artificial clove seeds |
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