CN107683768B - A kind of acclimatization and transplants method of the syringa reticulata var mandshurica tissue-cultured seedling of callus induction - Google Patents
A kind of acclimatization and transplants method of the syringa reticulata var mandshurica tissue-cultured seedling of callus induction Download PDFInfo
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- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
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- A—HUMAN NECESSITIES
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- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
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Abstract
The present invention provides a kind of acclimatization and transplants methods of the syringa reticulata var mandshurica tissue-cultured seedling of callus induction, belong to plant tissue culture technical field.The acclimatization and transplants method of the syringa reticulata var mandshurica tissue-cultured seedling of callus induction provided by the invention includes: that the syringa reticulata var mandshurica tissue-cultured seedling of callus induction is taken to carry out 5~10d of hardening, seedling after hardening, the temperature of the hardening is 15~18 DEG C, intensity of illumination is 800~1000lx, 5~8h of daily illumination, hardening humidity are 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 acclimatization and transplants process of the invention, the transplanting survival rate of the syringa reticulata var mandshurica tissue-cultured seedling of callus induction is up to 91% or more, acclimatization and transplants method provided by the invention is easy 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 fields, are more particularly to a kind of syringa reticulata var mandshurica group of callus induction
Train the acclimatization and transplants method of seedling.
Background technique
Syringa reticulata var mandshurica Syringareticulata (Blume), also known as Syringa amurensis, white lilac fall 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 25 kinds or more.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 in northern China gardens using relatively universal as a kind of ornamental plant
One of tree species, in addition, the solid densification of its material, structure are uniform；Essential oil content is abundant, 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 cuttage, the asexual process such as grafting.But seed used in seminal propagation needs complicated pregermination procedure, and emerges not
Uniformly, primary growth is slow and solid the features such as being influenced by biennial bearing, so that the sight flower type that this veriety of cloves is more, due to
It itself is mostly heterozygosis genotype, therefore seminal propagation cannot keep the typical character of its kind.
Currently, have certain research to syringa reticulata var mandshurica in vitro culture, it is true using bud proliferation 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 sterile wet tissue culture ambient condition
Under, to moisture around, sufficient, nutrient environment abundant produces adaptability to root system, 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 withered,
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, keeps survival rate lower.
Summary of the invention
In view of this, the purpose of the present invention is to provide a kind of shiftings of the hardening of the syringa reticulata var mandshurica tissue-cultured seedling of callus induction
Cultivating method reduces the transplanting death rate, improves survival rate.
In order to achieve the above-mentioned object of the invention, the present invention the following technical schemes are provided:
The present invention provides a kind of acclimatization and transplants methods of the syringa reticulata var mandshurica tissue-cultured seedling of callus induction, comprising:
1) it takes the syringa reticulata var mandshurica tissue-cultured seedling of callus induction to carry out 5~10d of hardening, obtains seedling after hardening, the hardening
Temperature be 15~18 DEG C, intensity of illumination be 800~1000lx, daily 5~8h of illumination, hardening humidity be 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 the mixture of one of river sand and vermiculite He turfy soil；
The nutrient solution includes 2mg/L's using 1/3MS fluid nutrient medium and 1/3WPM fluid nutrient medium as minimal medium
The NAA of IBA and 1mg/L.
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, comprising the following steps:
A) take the development hypocotyl of syringa reticulata var mandshurica be seeded on callus inducing medium carry out Fiber differentiation 40~
60d obtains callus；
B) the obtained callus of the step a) is inoculated on adventitious bud induction culture base, cultivates 40~55d, obtains more
The syringa reticulata var mandshurica tissue-cultured seedling of injured tissue induction.
Preferably, the culture medium of the step a) Fiber differentiation further includes using MS solid medium as minimal medium
The IBA of the BA and 0.5mg/L of 5mg/L.
Preferably, the condition of the step a) Fiber differentiation are as follows: 20~24 DEG C of temperature, intensity of illumination 2000lx, when illumination
Between daily 10~12h.
Preferably, the acquisition methods of the development hypocotyl are consolidated the following steps are included: the seed after disinfection is inoculated in MS
After carrying out in vitro culture 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 disinfection
75% ethyl alcohol impregnates 30s, aseptic water washing 3 times, after air-drying again with the mercuric chloride disinfection 5 of mass percentage 0.1%~
10min, aseptic water washing 3~5 times.
Preferably, the syringa reticulata var mandshurica tissue-cultured seedling of the callus induction further includes moving the tissue-cultured seedling before hardening
It plants and carries out strong seedling culture 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 medium are minimal medium, and pH value is 5.8~6.5.
The present invention provides a kind of acclimatization and transplants methods of the syringa reticulata var mandshurica tissue-cultured seedling of callus induction.The present invention provides
Acclimatization and transplants method will obtain tissue-cultured seedling in sterile moist environment, by hardening, so that tissue-cultured seedling has one to external environment
Fixed buffer capacity improves resistance；During hardening, nutrient solution is added in the root Xiang Youmiao, forms one kind and is conducive to plant growth
External nutrition environment, guarantee the enough nutrients of the root absorption of test tube seedling and moisture, not only saved cost, also reduce transplanting
The death rate improves survival rate to 91% or more.
Specific embodiment
The present invention provides a kind of acclimatization and transplants methods of the syringa reticulata var mandshurica tissue-cultured seedling of callus induction, comprising:
1) it takes the syringa reticulata var mandshurica tissue-cultured seedling of callus induction to carry out 5~10d of hardening, obtains seedling after hardening, the hardening
Temperature be 15~18 DEG C, intensity of illumination be 800~1000lx, daily 5~8h of illumination, hardening humidity be 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 the mixture of one of river sand and vermiculite He turfy soil；
The nutrient solution includes 2mg/L's using 1/3MS fluid nutrient medium and 1/3WPM fluid nutrient medium as minimal medium
The NAA of IBA and 1mg/L.
The present invention preferably carries out the cultivation of tissue-cultured seedling before carrying out acclimatization and transplants.In the present invention, the syringa reticulata var mandshurica is cured
The breeding method of the tissue-cultured seedling of injured tissue induction, comprising the following steps:
A) take the development hypocotyl of syringa reticulata var mandshurica be seeded on callus inducing medium carry out Fiber differentiation 40~
60d obtains callus；
B) the obtained callus of the step a) is inoculated on adventitious bud induction culture base, cultivates 40~55d, obtains 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: the syringa reticulata var mandshurica seed after disinfection being inoculated on MS solid medium and carries out in vitro culture
It cuts 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 preferred are as follows: clean up the seed, clear water rinses 1~
Then 2h impregnates 30s with the ethyl alcohol that volumn concentration is 75%, aseptic water washing 3 times, uses mass percentage again after air-drying
0.1% mercuric chloride sterilizes 5~10min, and aseptic water washing 3~5 times.
After the seed sterilized, the present invention cuts the seed of disinfection, cuts hypocotyl, after obtaining hypocotyl, this hair
It is bright that preferably hypocotyl is placed in in vitro culture base and carries out hypocotyl in vitro culture before carrying out induction of callus, In
In the present invention, the culture medium of the in vitro culture is preferably MS solid medium, and the condition of the hypocotyl in vitro culture is preferred
Are as follows: 20~24 DEG C of cultivation temperature, intensity of illumination 2000lx, the daily 12h of light application time；The time of the in vitro culture is preferred
10d；After the hypocotyl developed.The hypocotyl of development is preferably cut into the section of 5~7mm by the present invention, and access callus lures
It leads and carries out induction of callus on culture medium, the callus inducing medium is preferably basic with MS solid medium
Culture medium further includes the IBA of the BA and 0.5mg/L of 5mg/L；The condition of the Fiber differentiation is preferred are as follows: and 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 obtains 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 preferably using 1/2MS solid medium as minimal medium, comprising 0.1mg/L 2,4-
The NAA of the BA and 0.5mg/L of D, 3mg/L；The condition of the culture are as follows: 20~24 DEG C of temperature, intensity of illumination 2000lx, when 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 medium and 1/6WPM solid medium are minimal medium, in the present invention, the 1/6MS solid medium and
The volume ratio of 1/6WPM solid medium 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, increases the survival rate of seedling after 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, every 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 after hardening after seedling, 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 basic culture with 1/3MS fluid nutrient medium and 1/3WPM fluid nutrient medium
The volume ratio of base, described/3MS fluid nutrient medium and 1/3WPM fluid nutrient medium is preferably (1~3): (1~2), more preferably
1:1；The NAA of the IBA and 1mg/L of 2mg/L are preferably comprised in the nutrient solution.In the present invention, the transplanting is preferably transplanted to
Seedling after the transplanting is preferably transplanted in the nutritive cube of diameter 12cm, in each nutritive cube.In the present invention, the shifting
The mixture that matrix includes one of river sand and vermiculite He turfy soil is planted, i.e., the described transplanting medium includes river sand and turfy soil
The mixture of mixture or 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 lower, purchaser
Just, and the good water-retaining property of turfy soil, but if matrix is all turfy soil, be easy to cause tissue-cultured seedling rotten, when be added river sand or
After vermiculite, the water penetration of turfy soil is improved, is conducive to the transplanting survival rate for improving tissue-cultured seedling.In the present invention, the transplanting
Matrix using preceding preferably through 200 DEG C of disinfection 30min of high temperature, then cooled to room temperature；Matrix of the present invention makes
Preferably matrix is irrigated with water with preceding, specifically reaches matrix and drips；
The addition of heretofore described nutrient solution is on 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 injected, 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, and more preferably 20ml/ times；In the present invention, described inject is preferably
Injection every morning, preferred 7:30 every morning injection；In the present invention, the injection is preferably continuity injection, institute
The time for stating injection is preferably 13~16d, more preferably 15d.The mode of injection of the present invention can be such that seedling root is formed
The environment of one opposite high nutrition, high humidity, is conducive to plant growth, and the injection rate of 20ml is unlikely to form excessively high high humidity environment
So as to cause rotten, the nutrient solution described in local use, it is ensured that the enough nutrients of the root absorption of test tube seedling and moisture, greatly
The death rate is reduced greatly, survival rate is made to reach 91% or more.
Below with reference to embodiment to a kind of hardening shifting of the syringa reticulata var mandshurica tissue-cultured seedling of callus induction provided by the invention
Cultivating method is described in detail, but they cannot be interpreted as limiting the scope of the present invention.
Embodiment 1
(1) seed of the mature syringa reticulata var mandshurica of selection, is cleaned up using washing powder, then rinses 1h with clear water again；It sets
Aseptic filter paper is placed on aseptic water washing 3 times in using the ethyl alcohol that volumn concentration is 75% to impregnate 30s on superclean bench
It is upper to air-dry, then 8min is sterilized for 0.1% mercuric chloride with mass percentage, it then uses aseptic water washing 3 times, seed after must sterilizing；
(2) seed sterilized is cut, hypocotyl is accessed in MS culture medium and is cultivated, condition of culture are as follows: cultivation temperature
It is 22 DEG C, intensity of illumination 2000lx, the daily 12h of light application time is cultivated 10 days, hypocotyl after must developing；
(3) it takes the hypocotyl of 5mm to be seeded in using MS solid medium as minimal medium, includes 5mg/LBA, 0.5mg/
It is 20 DEG C, intensity of illumination 2000lx, daily illumination 12h in temperature on the callus inducing medium of LIBA, culture 50
It, obtains callus；
(4) it is inoculated into fine and close, emerald green, loose callus is grown using 1/2MS culture medium as minimal medium, includes
There are 2 of 0.1mg/L, is 20 DEG C in temperature on the adventitious bud induction culture base of the NAA of the BA and 0.5mg/L of 4-D, 3mg/L, 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 medium and 1/6WPM solid medium 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) sterile, 15 DEG C of temperature, intensity of illumination 800lx, daily illumination 5h, under 70% environment of indoor humidity, by seedling
It is transplanted into the transplanting culture medium of river sand and turfy soil volume ratio 2:5, transplants hardening 5d, obtain seedling after hardening, during hardening,
It is daily basic culture with 1/3MS fluid nutrient medium and 1/3WPM fluid nutrient medium to the stromal surface injection apart from seedling 1cm
Base stops injection after the nutrient solution 20ml, 15d of the NAA of IBA and 1mg/L comprising 2mg/L, and transplanting survival rate is up to 91%.
Embodiment 2
(1) seed of the mature syringa reticulata var mandshurica of selection, is cleaned up using washing powder, then rinses 2h with clear water again；It sets
Aseptic filter paper is placed on aseptic water washing 3 times in using the ethyl alcohol that volumn concentration is 75% to impregnate 30s on superclean bench
It is upper to air-dry, then 8min is sterilized for 0.1% mercuric chloride with mass percentage, it then uses aseptic water washing 3 times, seed after must sterilizing；
(2) seed sterilized is cut, hypocotyl is accessed in MS culture medium and is cultivated, condition of culture are as follows: cultivation temperature
It is 20 DEG C, intensity of illumination 2000lx, the daily 12h of light application time is cultivated 10 days, hypocotyl after must developing；
(3) it takes the hypocotyl of 6mm to be seeded in using MS solid medium as minimal medium, includes 5mg/LBA, 0.5mg/
It is 20 DEG C, intensity of illumination 2000lx, daily illumination 12h in temperature on the callus inducing medium of LIBA, culture 40
It, obtains callus；
(4) it is inoculated into fine and close, emerald green, loose callus is grown using 1/2MS culture medium as minimal medium, includes
There are 2 of 0.1mg/L, is 22 DEG C in temperature on the adventitious bud induction culture base of the NAA of the BA and 0.5mg/L of 4-D, 3mg/L, 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 medium and 1/6WPM solid medium 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) sterile, 16 DEG C of temperature, intensity of illumination 900lx, daily illumination 5h, under 70% environment of indoor humidity, by seedling
It is transplanted into the transplanting culture medium of river sand and turfy soil volume ratio 3:7, transplants hardening 7d, obtain seedling after hardening, during hardening,
It is daily basic culture with 1/3MS fluid nutrient medium and 1/3WPM fluid nutrient medium to the stromal surface injection apart from seedling 1cm
Base stops injection after the nutrient solution 15ml, 13d of the NAA of IBA and 1mg/L comprising 2mg/L, and transplanting survival rate is up to 93%.
Embodiment 3
(1) seed of the mature syringa reticulata var mandshurica of selection, is cleaned up using washing powder, then rinses 1.5h with clear water again；
It is placed in the ethyl alcohol immersion 30s for being 75% with volumn concentration on superclean bench and is placed on sterile filter with aseptic water washing 5 times
It is air-dried on paper, then sterilizes 8min with mass percentage for 0.1% mercuric chloride, then used aseptic water washing 3 times, planted after must sterilizing
Son；
(2) seed sterilized is cut, hypocotyl is accessed in MS culture medium and is cultivated, condition of culture are as follows: cultivation temperature
It is 24 DEG C, intensity of illumination 2000lx, the daily 12h of light application time is cultivated 10 days, hypocotyl after must developing；
(3) it takes the hypocotyl of 7mm to be seeded in using MS solid medium as minimal medium, includes 5mg/LBA, 0.5mg/
It is 20 DEG C, intensity of illumination 2000lx, daily illumination 12h in temperature on the callus inducing medium of LIBA, culture 60
It, obtains callus；
(4) it is inoculated into fine and close, emerald green, loose callus is grown using 1/2MS culture medium as minimal medium, includes
There are 2 of 0.1mg/L, is 24 DEG C in temperature on the adventitious bud induction culture base of the NAA of the BA and 0.5mg/L of 4-D, 3mg/L, 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 medium and 1/6WPM solid medium 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；
It (6), 15 DEG C of temperature, intensity of illumination 1000lx, daily illumination 8h, will be young under 80% environment of indoor humidity sterile
Transplantation of seedlings enters in the transplanting culture medium of river sand and turfy soil volume ratio 2:5, transplants hardening 10d, obtains seedling after hardening, hardening process
In, it is daily basic training with 1/3MS fluid nutrient medium and 1/3WPM fluid nutrient medium to the stromal surface injection apart from seedling 1cm
Base is supported, stops injection after the nutrient solution 25ml, 16d of the NAA of IBA and 1mg/L comprising 2mg/L, transplanting survival rate is reachable
92.6%.
Reference examples
It selects tissue-cultured seedling identical with Examples 1 to 3 to carry out hardening, the sealed membrane of tissue culture bottle is removed, seedling is taken out
Clean up root system；It is avoiding direct sunlight, 16 ± 2 DEG C of temperature, in the environment of humidity 80%, is being transplanted into turfy soil matrix,
After 15 days, plant strain growth gesture is weaker, and leaf some yellowing are beaten listless, and whole growing state is weaker, and the survival rate for recording seedling is
45.6%.
As seen from the above embodiment, it is moved using the hardening of the syringa reticulata var mandshurica tissue-cultured seedling of callus induction provided by the invention
Cultivating method is significantly higher than comparative example, 91% or more transplanting survival rate average out in survival rate.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (7)
1. a kind of acclimatization and transplants method of the syringa reticulata var mandshurica tissue-cultured seedling of callus induction, comprising:
1) it takes the syringa reticulata var mandshurica tissue-cultured seedling of callus induction to carry out 5~10d of hardening, obtains seedling after hardening, the temperature of the hardening
Degree is 15~18 DEG C, and intensity of illumination is 800~1000lx, and daily 5~8h of illumination, hardening humidity is 70~80%；
2) small transplantation of seedlings is added into the matrix of seedling root daily into transplanting medium after the hardening for obtaining step 1)
Nutrient solution；
The transplanting medium includes the mixture of one of river sand and vermiculite He turfy soil；
The nutrient solution is using 1/3MS fluid nutrient medium and 1/3WPM fluid nutrient medium as minimal medium, the IBA comprising 2mg/L
With the NAA of 1mg/L；
The breeding method of the tissue-cultured seedling, comprising the following steps:
A) it takes the development hypocotyl of syringa reticulata var mandshurica to be seeded to progress 40~60d of Fiber differentiation on callus inducing medium, obtains
Callus；The culture medium of the step a) Fiber differentiation further includes the BA of 5mg/L using MS solid medium as minimal medium
With the IBA of 0.5mg/L；
B) the obtained callus of the step a) is inoculated on adventitious bud induction culture base, cultivates 40~55d, obtains callus group
Knit the syringa reticulata var mandshurica tissue-cultured seedling of induction；The adventitious bud induction culture base includes using 1/2MS solid medium as minimal medium
The NAA of the BA and 0.5mg/L of 2,4-D, the 3mg/L of 0.1mg/L；
The syringa reticulata var mandshurica tissue-cultured seedling of the callus induction is before hardening, further includes: the tissue-cultured seedling is transplanted to strong sprout training
It supports and carries out strong seedling culture on base, the time of the strong seedling culture is 7~12d, and the strong seedling culture base is with 1/6MS solid medium
It is minimal medium with 1/6WPM solid medium, pH value is 5.8~6.5.
2. acclimatization and transplants method according to claim 1, which is characterized 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 transplants method according to claim 1, which is characterized 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 transplants method according to claim 1, which is characterized in that the step 2) nutrient solution adds 13 altogether~
16d, the amount added daily are 15~25ml.
5. acclimatization and transplants method according to claim 1, which is characterized in that the condition of the step a) Fiber differentiation are as follows:
20~24 DEG C of temperature, intensity of illumination 2000lx, the daily 10~12h of light application time.
6. acclimatization and transplants method according to claim 1, which is characterized in that the acquisition side of step a) the development hypocotyl
Method the following steps are included: by the seed after disinfection be inoculated on MS solid medium carry out in vitro culture 10d after, cut and sent out
Educate hypocotyl.
7. acclimatization and transplants method according to claim 6, which is characterized in that the disinfection is to clean up the seed,
Clear water rinses 1~2h, then impregnates 30s with the ethyl alcohol that volumn concentration is 75%, aseptic water washing 3 times, uses again after air-drying
The mercuric chloride of mass percentage 0.1% sterilizes 5~10min, and aseptic water washing 3~5 times.
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