CN104542308B - Rapid callus reproduction method for dioscorea bulbifera - Google Patents
Rapid callus reproduction method for dioscorea bulbifera Download PDFInfo
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Abstract
The invention provides a rapid callus reproduction method for dioscorea bulbifera. The rapid callus reproduction method comprises the following steps: performing induction culture on dioscorea bulbifera callus by using a callus induction culture medium, thereby obtaining callus; performing callus differentiation culture on the callus by using an MS reproduction culture medium, thereby obtaining cluster shoots; performing cluster shoot strengthening culture on the obtained cluster shoots by using an MS strengthening culture medium, thereby obtaining healthy and strong plants; performing rooting culture on the healthy and strong plants by using an MS rooting culture medium, thereby finally obtaining complete plants, and achieving seedling hardening and transplanting of dioscorea bulbifera seedlings. By adopting the rapid callus reproduction method, the callus differentiation speed is increased, the growth speed of the cluster shoots is increased, and the rooting percentage is increased. By adopting the rapid callus reproduction method provided by the invention, the single shoot reproduction coefficient is up to 30-50 times, the rooting rate of the obtained tissue culture seedlings is 95% or greater, each plant has 5-8 roots of 3-5cm on average, and the survival rate is 98% or greater after the plants are transplanted to sand beds.
Description
Technical field
The present invention relates to technical field of plant propagation, it is more particularly related to a kind of air potato callus is quickly bred
Method.
Background technology
Air potato (Dioscoreaceae), Dioscoreaceae, yam.General medicinal its stem tuber, is mainly used in each section's first shape
Gland disease and kinds cancer.Main in production to carry out vegetative propagation using bulbil, bulbil winter dormancy (Miao Cunming) is long-term
Vegetative propagation causes kind of a sexual involution, yield. and quality declines year after year.It is in short supply air potato seedling can be solved currently with callus seedling
Problem.
Current either air potato or other plant species have quickly is bred using callus.It is this traditional
Callus quickly breeds action principle：After plant tissue is sterilized, carry out Fiber differentiation and obtain callus or bud
Body, then to callus or bud proliferation, then carry out culture of rootage, last hardening and transplanting.The different phase pair of breeding
Culture medium has different requirements, so needing the usage and consumption for targetedly specifying each reproductive stage hormone.But, tradition
Method whole reproductive process all use same culture medium, these culture mediums cannot meet callus breeding each rank
The specificity of section needs, so callus cannot be promoted more preferably to grow up faster.
6-BA, IAA, IBA and NAA are typically added in traditional culture medium, the different meeting of amount that these materials are added
Different effects are produced, and these materials are also different for the effect that different plants produces.Beef extract belongs to animal groups
The additive of culture medium is knitted, it provides nutrient for animal tissue's culture.Beef extract is applied to plant tissue culture media can not
Good effect can be produced, beef extract is particularly applied to the training of air potato callus by presently relevant experiment or fewer
Support, it is at present or blank.
In order to solve this defect, a kind of method for quickly breeding of semen momordicae tissue cultures is disclosed in prior art, its
By using different culture medium and condition of culture in callus induction, callus differentiation and root induction stage, to promote
Enter the rooting rate of callus.
But by this method, it is propagation method for semen momordicae, due to having otherness between plant species, this
The method of kind is not particularly suited for air potato, and the technical scheme uses beef extract.Therefore invent a kind of new air potato callus into
Seedling method for quickly breeding becomes the problem of a urgent need to resolve.
The content of the invention
As the result of various extensive and careful research and experiment, it has been found by the inventor that healing in air potato
When the different phase of injured tissue seedling breeding uses the culture medium of NAA, KT, 6-BA and IAA for adding different content, these add
The compound for entering is favorably improved callus differentiation and the speed bred, and improves rooting rate, present inventor have further discovered that
Appropriate beef extract is added in each culture medium, the growth of callus can be promoted, make the more preferable of Multiple Buds growth.Base
In this discovery, the present invention is completed.
It is an object of the invention to solve at least the above, and provide the advantage that at least will be described later.
A further object of the invention is to obtain Multiple Buds in callus tissue culture, callus differentiation quickly to breed training
Support, Multiple Buds strong seedling culture and plant culture of rootage stage use respectively calli induction media, MS propagating culture mediums, MS strong sprouts
Culture medium and MS root medias, improve callus differentiation speed, improve the speed of growth of Multiple Buds, improve rooting rate.
A further object of the invention is that appropriate beef extract is added in each culture medium, promotes the life of callus
It is long, grow Multiple Buds more preferably.
A further object of the invention is that air potato callus tissue culture is quickly bred by biotechnology, improves air potato kind
The breeding coefficient and seedling quality of seedling, accomplishes scale production, and meets the needs in production.
In order to realize these purposes of the invention and further advantage, there is provided a kind of air potato callus seedling is quickly bred
Method, including：
Air potato induction of callus is carried out using calli induction media, callus is obtained；Trained using MS breedings
Foster base carries out callus differentiation culture to the callus of gained, obtains Multiple Buds；Using in MS strong seedling culture bases to gained
Multiple Buds carry out Multiple Buds strong seedling culture, obtain healthy and strong plant；The healthy and strong plant of gained is carried out using MS root medias
Plant culture of rootage finally gives whole plant, realizes hardening and the transplanting of air potato seedling；
Wherein, contain in the calli induction media：The beef extract of 4-6g/L, TDZ, 0.5- of 1.0-2.0mg/L
The sucrose of NAA, 25-30g/L of 1.0mg/L and the agar of 4.5-5.0g/L；
Contain in MS propagating culture mediums：The 6- of the beef extract of 4-6g/L, KT, 1.0-2.0mg/L of 0.1-0.4mg/L
The sucrose of GA3,25-30g/L of IBA, 1-5g/L of BA, 0.2-1.0mg/L and the agar of 4.5-5.0g/L；
Contain in MS strong seedling culture bases：The beef extract of 4-6g/L, 6-BA, 0.1-0.4mg/L of 0.5-1.5mg/L
The sucrose of IAA, 25-30g/L of KT, 0.2-0.6mg/L and the agar of 4.5-5.0g/L；
Contain in MS root medias：The beef extract of 4-6g/L, the IBA of NAA, 0.5-1.0mg/L of 1.0-2.0mg/L,
The sucrose of 15-30g/L and the agar of 4.5-5.0g/L.
Preferably, in described air potato callus seedling forming quick propagating method, the method for the callus tissue culture is：Will
In vitro cuttings cut into section and are inoculated in calli induction media, are 23-26 DEG C in cultivation temperature, intensity of illumination 1400-
2000lux, light application time is that culture obtains callus in 24-25 days under conditions of 10-12 hours/day.
Preferably, in described air potato callus seedling forming quick propagating method, callus differentiation and cultivation process is：By institute
The callus for obtaining is inoculated in MS propagating culture mediums, is 23-26 DEG C in cultivation temperature, intensity of illumination 1400-2000lux, light
It is to cultivate 29-30 days under conditions of 10-12 hours/day according to the time, to lateral bud redifferentiation Multiple Buds is obtained.
Preferably, in described air potato callus seedling forming quick propagating method, the Multiple Buds strong seedling culture method is：Will
The Multiple Buds of gained are placed in MS strong seedling culture bases, in cultivation temperature 23-26 DEG C, intensity of illumination 1400-2000lux, during illumination
Between obtain within 19-20 days healthy and strong plant for culture under conditions of 10-12 hours/day.
Preferably, in described air potato callus seedling forming quick propagating method, the plant culture of rootage step includes：Will
The healthy and strong plant of gained is placed in MS root medias, in cultivation temperature 23-26 DEG C, intensity of illumination 1400-2000lux, illumination
Time is that culture obtains the whole plant with root for 39-40 days under conditions of 10-12 hours/day.
Preferably, in described air potato callus seedling forming quick propagating method, the hardening and transplanting are comprised the following steps：
Step one, by the whole plant of gained, be under the conditions of 22-25 DEG C, using water hardening 2-4 days, surface angle in temperature
Matter is taken out after being formed, and obtains seedling strain；
Step 2, gained seedling strain is transplanted to well-ventilated and intensity of illumination for the sandy soil earth of 500-2000lux, in sand
Little tree is obtained after cultivating 30-50 days in soil；
Step 3, gained little tree is transplanted into land for growing field crops, transplanted in latter week, daily spraying 3-5 time, each 10min；One
Spraying 1-2 time daily, each 10min after week；Temperature conditionss during transplanting are 20-28 DEG C, relative humidity 75-80%, sunshade
Rate is 65-75%, finally obtains transplanting seedling.
Preferably, in described air potato callus seedling forming quick propagating method, the initial pH of each culture medium is 5-
6。
The present invention at least includes following beneficial effect：First, the 6-BA of 1.0-2.0mg/L is added in MS propagating culture mediums
With the quick differentiation that the kinetin KT of 0.1-0.4mg/L promotes Multiple Buds；Addition concentration promotes for the IBA of 0.2-1.0mg/L
The growth of Multiple Buds；
Add concentration in MS strong seedling culture bases and promote clump for the IAA of the 6-BA and 0.2-0.6mg/L of 0.5-1.5mg/L
Sprout the rapid deployment of blade；
The NAA of IBA and 1.0-2.0mg/L that concentration is 0.5-1.0mg/L is applied in combination on MS root medias, is promoted
Multiple Buds fast-growth becomes the whole plant with root, and these plant can directly transplant sand bed Jing after hardening.
Secondly, the present invention adds the beef extract of 4-6g/L in each culture medium, can promote air potato callus
Growth, make the more preferable of Multiple Buds length.
Again, air potato callus tissue culture is quickly bred by biotechnology, cultivates be available in a large number at short notice
The air potato seedling of field production, improves the breeding coefficient and seedling quality of air potato seedling, accomplishes scale production, and meets production
On needs.
Finally, method for quickly breeding of the present invention, the simple bud growth coefficient for obtaining reaches 30-50 times, the group of acquisition
Training seedling rooting rate more than 95%, every plant of average band 5-8 root, root is a length of 3-5 centimetre, transplanting sand bed survival rate 98% with
On.
The further advantage of the present invention, target and feature embody part by description below, and part will also be by this
The research of invention and practice and be understood by the person skilled in the art.
Specific embodiment
With reference to example, the present invention is described in further detail, to make those skilled in the art with reference to specification text
Word can be implemented according to this.
A kind of air potato callus seedling forming quick propagating method, including：
Air potato induction of callus is carried out using calli induction media, callus is obtained；Trained using MS breedings
Foster base carries out callus differentiation culture to the callus of gained, obtains Multiple Buds；Using in MS strong seedling culture bases to gained
Multiple Buds carry out Multiple Buds strong seedling culture, obtain healthy and strong plant；The healthy and strong plant of gained is carried out using MS root medias
Plant culture of rootage finally gives whole plant, realizes hardening and the transplanting of air potato seedling；
Wherein, contain in the calli induction media：The beef extract of 4-6g/L, TDZ, 0.5- of 1.0-2.0mg/L
The sucrose of NAA, 25-30g/L of 1.0mg/L and the agar of 4.5-5.0g/L；
Contain in MS propagating culture mediums：The 6- of the beef extract of 4-6g/L, KT, 1.0-2.0mg/L of 0.1-0.4mg/L
The sucrose of GA3,25-30g/L of IBA, 1-5g/L of BA, 0.2-1.0mg/L and the agar of 4.5-5.0g/L；
Contain in MS strong seedling culture bases：The beef extract of 4-6g/L, 6-BA, 0.1-0.4mg/L of 0.5-1.5mg/L
The sucrose of IAA, 25-30g/L of KT, 0.2-0.6mg/L and the agar of 4.5-5.0g/L；
Contain in MS root medias：The beef extract of 4-6g/L, the IBA of NAA, 0.5-1.0mg/L of 1.0-2.0mg/L,
The sucrose of 15-30g/L and the agar of 4.5-5.0g/L.
Beef extract directly can not only provide nutrition and the energy for air potato, and be conducive to the life of probio in culture medium
Long, probio can promote the growth and development of air potato, and beef extract decomposition is obtained carbon source, nitrogen source, phosphate by probio
And vitamin, it is connected in air potato and nutrient is provided.
In described air potato callus seedling forming quick propagating method, the method for the callus tissue culture is：By sterile test tube
Seedling cuts into section and is inoculated in calli induction media, is 23-26 DEG C in cultivation temperature, intensity of illumination 1400-2000lux, light
It is that culture obtains callus in 24-25 days under conditions of 10-12 hours/day according to the time.
In described air potato callus seedling forming quick propagating method, callus differentiation and cultivation process is：By the callus of gained
Tissue is inoculated in MS propagating culture mediums, is 23-26 DEG C in cultivation temperature, intensity of illumination 1400-2000lux, and light application time is
Cultivate 29-30 days under conditions of 10-12 hours/day, to lateral bud redifferentiation Multiple Buds are obtained.
In described air potato callus seedling forming quick propagating method, the Multiple Buds strong seedling culture method is：By the clump of gained
Sprout and be placed in MS strong seedling culture bases, in cultivation temperature 23-26 DEG C, intensity of illumination 1400-2000lux, light application time is 10-12
Culture under conditions of hour/day obtains healthy and strong plant for 19-20 days.
In described air potato callus seedling forming quick propagating method, the plant culture of rootage step includes：By the strong of gained
Strong plant is placed in MS root medias, and in cultivation temperature 23-26 DEG C, intensity of illumination 1400-2000lux, light application time is 10-
Culture under conditions of 12 hours/day obtains the whole plant with root for 39-40 days.
In described air potato callus seedling forming quick propagating method, the hardening and transplanting are comprised the following steps：
Step one, by the whole plant of gained, be under the conditions of 22-25 DEG C, using water hardening 2-4 days, surface angle in temperature
Matter is taken out after being formed, and obtains seedling strain；
Step 2, gained seedling strain is transplanted to well-ventilated and intensity of illumination for the sandy soil earth of 500-2000lux, in sand
Little tree is obtained after cultivating 30-50 days in soil；
Step 3, gained little tree is transplanted into land for growing field crops, transplanted in latter week, daily spraying 3-5 time, each 10min；One
Spraying 1-2 time daily, each 10min after week；Temperature conditionss during transplanting are 20-28 DEG C, relative humidity 75-80%, sunshade
Rate is 65-75%, finally obtains transplanting seedling.
In described air potato callus seedling forming quick propagating method, the initial pH of each culture medium is 5-6.
Example 1
A kind of air potato callus tissue culture method for quickly breeding, comprises the following steps：
(1) induction of callus：The stem section without bud that in vitro cuttings cut into 1cm length is inoculated into into callus induction
It it is 23 DEG C in cultivation temperature in culture medium, intensity of illumination 1400lux, light application time is to cultivate to obtain for 25 days under conditions of 10h/d
Callus.Add 5g/L beef extracts, 1.0mg/L TDZ, 0.5mg/LNAA, 30g/L sugarcane wherein in calli induction media
The agar of sugar and 5.0g/L, the initial pH value of culture medium is 5.8.
(2) callus differentiation culture：The callus that step (1) is obtained is inoculated in MS propagating culture mediums, in training
Foster temperature is 23 DEG C, intensity of illumination 1400lux, and light application time is to cultivate 30 days under conditions of 10h/d, is obtained after callus differentiation
A large amount of Multiple Buds are obtained, 5g/L beef extracts, 0.2mg/L KT, 1mg/L 6-BA, 0.2mg/L are added wherein in MS propagating culture mediums
The agar of the GA3 of IBA, 1g/L, 30g/L sucrose and 5.0g/L, the initial pH value of culture medium is 5.8.
(3) Multiple Buds strong seedling culture：The Multiple Buds obtained in step (2) are placed in MS strong seedling culture bases and are cultivated 30 days
To healthy and strong plant, 23 DEG C of cultivation temperature, intensity of illumination 1400lux, light application time is 12 hours/day；Wherein MS strong seedling cultures base
In beef extract containing 5g/L, 0.5mg/L 6-BA, 0.2mg/L IAA, the agar of the KT of 0.2mg/L, 30g/L sucrose and 5g/L,
The initial pH value of culture medium is 5.8.
(4) rooting of vitro seedling culture：The Multiple Buds obtained in step (3) are placed in MS root medias and are cultivated 20 days
To the whole plant with root, 23 DEG C of cultivation temperature, intensity of illumination 1400lux, light application time is 12 hours/day；Wherein MS is taken root
The agar of 5g/L containing beef extract in culture medium, 1mg/L NAA, the IBA of 0.5mg/L, 15g/L sucrose and 5g/L, culture medium
Initial pH value is 5.8.
(5) hardening and transplanting：After obtaining the whole plant with root, in the indoor opening bottle cap that room temperature is 25 DEG C or so,
A small amount of running water is added in bottle, hardening 2 days, surface horny takes out seedling after being formed, and cleans root culture medium, is transplanted to immediately logical
Wind grows after one and a half months well and in the sandy soil earth of the low light level, in sandy soil earth transplants land for growing field crops.Transplant in latter week, daily
Early 8 points to late 6 points are sprayed 3 times, each 8-10min, hereafter early 8 points daily, late 6 points respectively spraying 1 times, each 10min；During transplanting
Temperature conditionss are 20 DEG C, relative humidity 75-80%, sunshade rate 70%.
Example 2
A kind of air potato callus tissue culture method for quickly breeding, comprises the following steps：
(1) induction of callus：The stem section without bud that in vitro cuttings cut into 1cm length is inoculated into into callus induction
It it is 24 DEG C in cultivation temperature in culture medium, intensity of illumination 1500lux, light application time is to cultivate to obtain for 25 days under conditions of 11h/d
Callus.Add 5.5g/L beef extracts, 1.2mg/L TDZ, 0.6mg/LNAA, 30g/L wherein in calli induction media
The agar of sucrose and 5.0g/L, the initial pH value of culture medium is 5.8.
(2) callus differentiation culture：The callus that step (1) is obtained is inoculated in MS propagating culture mediums, in training
Foster temperature is 24 DEG C, intensity of illumination 1500lux, and light application time is to cultivate 30 days under conditions of 11h/d, is obtained after callus differentiation
A large amount of Multiple Buds are obtained, addition 5.5g/L beef extracts wherein in MS propagating culture mediums, 0.4mg/L KT, 1.5mg/L 6-BA,
The agar of the GA3 of 0.5mg/L IBA, 1.5g/L, 30g/L sucrose and 5.0g/L, the initial pH value of culture medium is 5.8.
(3) Multiple Buds strong seedling culture：The Multiple Buds obtained in step (2) are placed in MS strong seedling culture bases and are cultivated 30 days
To healthy and strong plant, 24 DEG C of cultivation temperature, intensity of illumination 1500lux, light application time is 12 hours/day；Wherein MS strong seedling cultures base
In beef extract containing 5.5g/L, 1mg/L 6-BA, 0.4mg/L IAA, the agar of the KT of 0.3mg/L, 30g/L sucrose and 5g/L,
The initial pH value of culture medium is 5.8.
(4) rooting of vitro seedling culture：The Multiple Buds obtained in step (3) are placed in MS root medias and are cultivated 20 days
To the whole plant with root, 24 DEG C of cultivation temperature, intensity of illumination 1500lux, light application time is 12 hours/day；Wherein MS is taken root
The agar of the IBA of 5.5g/L containing beef extract in culture medium, 1.5mg/LNAA, 0.8mg/L, 15g/L sucrose and 5g/L, culture medium
Initial pH value be 5.8.
(5) hardening and transplanting：After obtaining the whole plant with root, in the indoor opening bottle cap that room temperature is 25 DEG C or so,
A small amount of running water is added in bottle, hardening 3 days, surface horny takes out seedling after being formed, and cleans root culture medium, is transplanted to immediately logical
Wind grows after one and a half months well and in the sandy soil earth of the low light level, in sandy soil earth transplants land for growing field crops.Transplant in latter week, daily
Early 8 points to late 6 points are sprayed 4 times, each 8-10min, hereafter early 8 points daily, late 6 points respectively spraying 2 times, each 10min；During transplanting
Temperature conditionss are 25 DEG C, relative humidity 75-80%, sunshade rate 70%.
Example 3
A kind of air potato callus tissue culture method for quickly breeding, comprises the following steps：
(1) induction of callus：The stem section without bud that in vitro cuttings cut into 1cm length is inoculated into into callus induction
It it is 25 DEG C in cultivation temperature in culture medium, intensity of illumination 2000lux, light application time is to cultivate to obtain for 25 days under conditions of 12h/d
Callus.Wherein in calli induction media add 6g/L beef extracts, 2mg/L TDZ, 1mg/L NAA, 30g/L sucrose and
The agar of 5.0g/L, the initial pH value of culture medium is 5.8.
(2) callus differentiation culture：The callus that step (1) is obtained is inoculated in MS propagating culture mediums, in training
Foster temperature is 25 DEG C, intensity of illumination 2000lux, and light application time is to cultivate 30 days under conditions of 12h/d, is obtained after callus differentiation
A large amount of Multiple Buds are obtained, 6g/L beef extracts, 0.1mg/L KT, 2mg/L 6-BA, 1mg/L are added wherein in MS propagating culture mediums
The agar of the GA3 of IBA, 5g/L, 30g/L sucrose and 5.0g/L, the initial pH value of culture medium is 5.8.
(3) Multiple Buds strong seedling culture：The Multiple Buds obtained in step (2) are placed in MS strong seedling culture bases and are cultivated 30 days
To healthy and strong plant, 25 DEG C of cultivation temperature, intensity of illumination 2000lux, light application time is 12 hours/day；Wherein MS strong seedling cultures base
In beef extract containing 6g/L, 1.5mg/L 6-BA, 0.6mg/L IAA, the agar of the KT of 0.4mg/L, 30g/L sucrose and 5g/L,
The initial pH value of culture medium is 5.8.
(4) rooting of vitro seedling culture：The Multiple Buds obtained in step (3) are placed in MS root medias and are cultivated 20 days
To the whole plant with root, 25 DEG C of cultivation temperature, intensity of illumination 2000lux, light application time is 12 hours/day；Wherein MS is taken root
At the beginning of the agar of 6g/L containing beef extract in culture medium, 2mg/L NAA, the IBA of 1mg/L, 15g/L sucrose and 5g/L, culture medium
Beginning pH value is 5.8.
(5) hardening and transplanting：After obtaining the whole plant with root, in the indoor opening bottle cap that room temperature is 25 DEG C or so,
A small amount of running water is added in bottle, hardening 4 days, surface horny takes out seedling after being formed, and cleans root culture medium, is transplanted to immediately logical
Wind grows after one and a half months well and in the sandy soil earth of the low light level, in sandy soil earth transplants land for growing field crops.Transplant in latter week, daily
Early 8 points to late 6 points are sprayed 4 times, each 8-10min, hereafter early 8 points daily, late 6 points respectively spraying 3 times, each 10min；During transplanting
Temperature conditionss are 25 DEG C, relative humidity 75-80%, sunshade rate 70%.
Although embodiment of the present invention is disclosed as above, it is not restricted to listed in specification and embodiment
With.It can be applied to completely various suitable the field of the invention.For those skilled in the art, can be easily
Realize other modification.Therefore under the universal limited without departing substantially from claim and equivalency range, the present invention is not limited
In specific details and shown here as the example with description.
Claims (7)
1. a kind of air potato callus seedling forming quick propagating method, it is characterised in that：
Air potato induction of callus is carried out using calli induction media, callus is obtained；Using MS propagating culture mediums
Callus differentiation culture is carried out to the callus of gained, Multiple Buds are obtained；Using the clump in MS strong seedling culture bases to gained
Sprouting carries out Multiple Buds strong seedling culture, obtains healthy and strong plant；Plant is carried out to the healthy and strong plant of gained using MS root medias
Culture of rootage finally gives whole plant, realizes hardening and the transplanting of air potato seedling；
Wherein, the calli induction media is：The beef extract of 4-6g/L, TDZ, 0.5-1.0mg/L of 1.0-2.0mg/L
The sucrose of NAA, 25-30g/L and the agar of 4.5-5.0g/L；
MS propagating culture mediums are：6-BA, 0.2- of the beef extract of MS, 4-6g/L, KT, 1.0-2.0mg/L of 0.1-0.4mg/L
The sucrose of GA3,25-30g/L of IBA, 1-5g/L of 1.0mg/L and the agar of 4.5-5.0g/L；
MS strong seedling culture bases are：KT, 0.2- of the beef extract of MS, 4-6g/L, 6-BA, 0.1-0.4mg/L of 0.5-1.5mg/L
The sucrose of IAA, 25-30g/L of 0.6mg/L and the agar of 4.5-5.0g/L；
MS root medias are：IBA, 15- of the beef extract of MS, 4-6g/L, NAA, 0.5-1.0mg/L of 1.0-2.0mg/L
The sucrose of 30g/L and the agar of 4.5-5.0g/L.
2. air potato callus seedling forming quick propagating method as claimed in claim 1, it is characterised in that the callus induction training
Foster method is：In vitro cuttings are cut into into section to be inoculated in calli induction media, is 23-26 DEG C in cultivation temperature, light
According to intensity 1400-2000lux, light application time is that culture obtains callus in 24-25 days under conditions of 10-12 hours/day.
3. air potato callus seedling forming quick propagating method as claimed in claim 1, it is characterised in that callus breaks up culture side
Method is：The callus of gained is inoculated in MS propagating culture mediums, is 23-26 DEG C in cultivation temperature, intensity of illumination 1400-
2000lux, light application time is to cultivate 29-30 days under conditions of 10-12 hours/day, and to lateral bud redifferentiation Multiple Buds are obtained.
4. air potato callus seedling forming quick propagating method as claimed in claim 1, it is characterised in that the Multiple Buds strong seedling culture
Method is：The Multiple Buds of gained are placed in MS strong seedling culture bases, in cultivation temperature 23-26 DEG C, intensity of illumination 1400-
2000lux, light application time is that culture obtains healthy and strong plant for 19-20 days under conditions of 10-12 hours/day.
5. air potato callus seedling forming quick propagating method as claimed in claim 1, it is characterised in that the plant culture of rootage step
Suddenly include：The healthy and strong plant of gained is placed in MS root medias, in cultivation temperature 23-26 DEG C, intensity of illumination 1400-
2000lux, light application time is that culture obtains the whole plant with root for 39-40 days under conditions of 10-12 hours/day.
6. the air potato callus seedling forming quick propagating method as described in claim 1 or 5, it is characterised in that the hardening and transplanting
Comprise the following steps：
Step one, by the whole plant of gained, be under the conditions of 22-25 DEG C, using water hardening 2-4 days, surface horny shape in temperature
Into rear taking-up, seedling strain is obtained；
Step 2, gained seedling strain is transplanted to well-ventilated and intensity of illumination for the sandy soil earth of 500-2000lux, in sandy soil earth
Middle culture obtains little tree after 30-50 days；
Step 3, gained little tree is transplanted into land for growing field crops, transplanted in latter week, daily spraying 3-5 time, each 10min；One week
Spray 1-2 time daily afterwards, each 10min；Temperature conditionss during transplanting are 20-28 DEG C, relative humidity 75-80%, and sunshade rate is
65-75%, finally obtains transplanting seedling.
7. air potato callus seedling forming quick propagating method as claimed in claim 1, it is characterised in that each culture medium it is initial
PH is 5-6.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510056427.8A CN104542308B (en) | 2015-02-03 | 2015-02-03 | Rapid callus reproduction method for dioscorea bulbifera |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510056427.8A CN104542308B (en) | 2015-02-03 | 2015-02-03 | Rapid callus reproduction method for dioscorea bulbifera |
Publications (2)
| Publication Number | Publication Date |
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