CN108719062A - A kind of quick breeding method for tissue culture of brussels sprout - Google Patents
A kind of quick breeding method for tissue culture of brussels sprout Download PDFInfo
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- CN108719062A CN108719062A CN201810445577.1A CN201810445577A CN108719062A CN 108719062 A CN108719062 A CN 108719062A CN 201810445577 A CN201810445577 A CN 201810445577A CN 108719062 A CN108719062 A CN 108719062A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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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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Abstract
The present invention configures the culture mediums for adventitious bud inducing of suitable brussels sprout tissue cultures, the culture medium for shoot proliferation and for the culture medium of sterile rootage according to the growth characteristics of brussels sprout on the basis of MS culture mediums.Breeding coefficient can be effectively improved using the culture medium of the present invention for adventitious bud inducing, the culture medium for shoot proliferation and the culture medium for sterile rootage, reduces production cost；With short production cycle, planting benefit greatly improves；Also technical support is provided for the molecular breeding of brussels sprout.
Description
Technical field
The present invention relates to a kind of quick breeding method for tissue culture of brussels sprout, belong to micropropagation of plants technology neck
Domain.
Background technology
Brussels sprout is also referred to as Brussels sprouts and son holds wild cabbage, originates in Mediterranean, belongs to Cruciferae Brassica genus wild cabbage
Kind biennial herb plant is one of the important vegetables of Europe and North American countries.Brussels sprout is the mutation of brassica specie, because of it
Middle axillary bud can form small leaf-head and gain the name for edible.The flavor of brussels sprout is like cabbage but also has itself unique mouth
Taste, small leaf-head nutritive value is very high, and the content of protein occupies first of cabbage vegetable, and rich in vitamin C, selenium, carrot
Element, vitamin B1 and vitamin B2, wherein protein content is highest in cabbage vegetable.Brussels sprout is introduced at the end of the 20th century
To China and start to plant, makes popular vegetables.Although introducing the time not grow, the approval in market is obtained, very
Prevalence, often supply falls short of demand in the market.Mainly there are 2 kinds of brussels sprouts in China market, one is purples, another is green
Color.
The research of wild cabbage is more, has been successfully established tissue culture technique system.But brussels sprout is relevant at present
Study it is less, it is very weak.The tissue culture technique of brussels sprout is not yet reported that, also without carrying out brussels sprout factory metaplasia
Production.Tissue culture technique can largely provide the seedling of brussels sprout in a short time, carry out the quick breeding of good seed.
Brassica oleracea var gemmifera belongs to cenospecies, and breeding cycle is long, and characters of progenies can be easily separated, the fixation and tissue training of merit
It supports compared to very difficult, the character of the elite plant strain of filial generation selection and breeding can be carried out by tissue cultures vegetative propagation quick
It is fixed.
For above present situation, the present invention provides a kind of quick breeding method for tissue culture of brussels sprout, can be fast
Speed breeding good seed, improves plantation and the income of sellers, promotes the development of brussels sprout industry.
Invention content
It is weak it is an object of the invention to solve current brussels sprout seedling research, quickly to breed brussels sprout seedling,
A kind of quick, low cost, efficient quick breeding method for tissue culture are provided.
To achieve the above object, the technical method that the present invention uses is as follows：
A kind of quick breeding method for tissue culture of brussels sprout, includes the following steps：(1) explant is chosen and is sterilized,
(2) preparation of culture medium, (3) adventitious bud inducing, (4) shoot proliferation, (5) sterile rootage and (6) hardening and transplanting.This method
It is as follows：
(1) explant is chosen and is sterilized：
1. choosing the tender branch of brussels sprout of healthy growth as explant；
2. washing branch with neutral detergent, to remove the dirt on branch surface, extra blade is then removed, totally
Paper handkerchief blots surface moisture；
3. on superclean bench, first 10-30s is impregnated with 70% alcohol, then with aseptic water washing 3 times, each 3-5min；
5-8min is sterilized 4. being transferred to immediately in the mercuric chloride solution of 0.1-0.3%, finally with sterile water washing 3-5 times, every time
3-5min, be sterilized explant.
(2) preparation of culture medium：
1. the preparation of modified MS medium, is formulated and is：The CaCl2 in original MS culture mediums is replaced with Ca (NO3) 24H2O
2H2O simultaneously adds riboflavin, and other components are unchanged；A concentration of 50- of Ca (NO3) 24H2O in modified MS medium
100mg/L, a concentration of 0.3-5mg/L of riboflavin；
2. preparing the culture medium for adventitious bud inducing, it is formulated and is：In the modified MS medium add 6-BA, KT with
And IAA；A concentration of 0.5-0.8mg/L of 6-BA in adventitious bud induction culture base, a concentration of 0.5-1mg/L of KT, IAA's is dense
Degree is 0.05-0.5mg/L；
3. preparing the culture medium for shoot proliferation, it is formulated and is：On the basis of the modified MS medium add 6-BA,
KT and IAA, a concentration of 0.5-2mg/L of 6-BA in subculture multiplication medium, a concentration of 0.1-2mg/L of KT, IAA's is dense
Degree is 0.05-5mg/L；
4. preparing the culture medium for sterile rootage, it is formulated and is：On the basis of the modified MS medium add IBA and
IAA, a concentration of 0.1-1mg/L of IBA, a concentration of 0.5-5mg/L of IAA in root media.
(3) adventitious bud inducing：
The disinfection explant that step (1) obtains is cut into the simple bud of 0.4-0.5cm long or double leaf stem sections, is then inoculated in
For the culture medium of adventitious bud inducing, Fiber differentiation is carried out 18-22 days, obtain brussels sprout adventitious bud；Inducing culturing condition is such as
Under：Temperature is 26 ± 3 DEG C, humidity 80-90%, and light application time is 14 hours/day, intensity of illumination 800-12001x.
(4) shoot proliferation：
Culture medium of the brussels sprout adventitious bud access that step (3) is obtained for shoot proliferation, carries out squamous subculture 20-
25 days, obtain brussels sprout aseptic seedling；Shoot proliferation condition of culture is as follows：Temperature is 26 ± 3 DEG C, humidity 50-65%, illumination
Time is 14 hours/day, intensity of illumination 800-12001x；
(5) sterile rootage：
The brussels sprout aseptic seedling that step (4) obtains cut numerous, and is kept for every plant there are one adventitious bud, access is used
Culture of rootage is carried out in the culture medium of sterile rootage 15-20 days, obtain brussels sprout seedling；Culture of rootage condition is as follows：Training
It is 26 ± 3 DEG C, humidity 50-65% to support temperature, and light application time is 14 hours/day, intensity of illumination 800-12001x；
In step (5) after brussels sprout growth of seedling to root long 0.5cm, height of seedling 3cm, culture bottle cap is opened, it is indoor
Then root culture medium is cleaned in hardening culture 5 days, move into the hole tray using 64 holes containing the peat soil after disinfection, it is big to be placed in plastics
Canopy keeps 85% or more humidity.
Specifically, Ca (NO in modified MS medium3)2·4H2A concentration of 60mg/L of O, a concentration of 2.5mg/ of riboflavin
L。
Specifically, in the culture medium of adventitious bud inducing 6-BA a concentration of 1mg/L, a concentration of 0.5mg/L of KT, IAA's
A concentration of 0.2mg/L.
Specifically, in the culture medium of shoot proliferation 6-BA a concentration of 2mg/L, a concentration of 0.5mg/L of KT.
Specifically, in the culture medium of sterile rootage IBA a concentration of 0.5mg/L, a concentration of 1mg/L of IAA.
The beneficial effects of the invention are as follows：
Tissue-culturing quick-propagation is carried out to the axillary bud of brussels sprout stem section using culture medium of the present invention, is effectively improved
Breeding coefficient, fixed merit, also can effectively produce brassica oleracea var gemmifera seedling, also provide good economic effect for grower
Benefit.
Description of the drawings
Fig. 1 is the schematic diagram of brussels sprout of the present invention (purple) tissue-culturing quick-propagation, wherein a is purple
Brussels sprout adventitious bud inducing, b are that purple brussels sprout takes root, and c plants for purple brussels sprout tissue-cultured seedling.
Fig. 2 is the schematic diagram of brussels sprout of the present invention (green) tissue-culturing quick-propagation, wherein a is green
Brussels sprout is proliferated, and b is that green brussels sprout takes root, and c is green brussels sprout tissue-cultured seedling plantation.
Specific implementation mode
Embodiment 1
A kind of quick breeding method for tissue culture of purple brussels sprout, includes the following steps：
(1) explant is chosen and is sterilized：
1. choosing the tender branch of brussels sprout of healthy growth as explant；
2. washing branch with neutral detergent, to remove the dirt on branch surface, extra blade is then removed, totally
Paper handkerchief blots surface moisture；
3. on superclean bench, first 10-30s is impregnated with 70% alcohol, then with aseptic water washing 3 times, each 3-5min；
5-8min is sterilized 4. being transferred to immediately in the mercuric chloride solution of 0.1-0.3%, finally with sterile water washing 3-5 times, every time
3-5min, be sterilized explant.
(2) preparation of modified MS medium：
The formula of modified MS medium is：With Ca (NO3)2·4H2O replaces the CaCl in original MS culture mediums2·2H2O simultaneously adds
Add riboflavin, other components unchanged；Wherein Ca (NO3)2·4H2A concentration of 60mg/L of O, a concentration of 5mg/ of riboflavin
L。
(3) adventitious bud inducing：
The disinfection explant that step (1) obtains is cut into the simple bud of 0.4-0.5cm long or double leaf stem sections, is then inoculated in
For the culture medium of adventitious bud inducing, Fiber differentiation is carried out 18-22 days, obtain brussels sprout adventitious bud；Inducing culturing condition is such as
Under：Temperature is 26 ± 3 DEG C, humidity 80-90%, and light application time is 14 hours/day, intensity of illumination 800-12001x；
The formula of the culture medium of adventitious bud inducing is：6-BA, KT and IAA are added in the modified MS medium；Its
A concentration of 0.2mg/L of a concentration of 0.6mg/L of a concentration of 0.8mg/L of middle 6-BA, KT, IAA.
(4) shoot proliferation：
Culture medium of the brussels sprout adventitious bud access that step (3) is obtained for shoot proliferation, carries out squamous subculture 20-
25 days, obtain brussels sprout aseptic seedling；Shoot proliferation condition of culture is as follows：Temperature is 26 ± 3 DEG C, humidity 50-65%, illumination
Time is 14 hours/day, intensity of illumination 800-12001x；
The formula of culture medium for shoot proliferation is：On the basis of the modified MS medium add 6-BA, KT with
And a concentration of 0.5mg/L of a concentration of 1mg/L of IAA, wherein 6-BA, KT, a concentration of IAA0.2mg/L.
(5) sterile rootage：
The brussels sprout aseptic seedling that step (4) obtains cut numerous, and is kept for every plant there are one adventitious bud, access is used
Culture of rootage is carried out in the culture medium of sterile rootage 15-20 days, obtain brussels sprout seedling；Culture of rootage condition is as follows：Training
It is 26 ± 3 DEG C, humidity 50-65% to support temperature, and light application time is 14 hours/day, intensity of illumination 800-12001x.
The formula of culture medium for sterile rootage is：IBA and IAA is added on the basis of the modified MS medium,
A concentration of 5mg/L of a concentration of 0.1mg/L of wherein IBA, IAA.
(6) hardening and transplanting：
In step (5) after brussels sprout growth of seedling to root long 0.5cm, height of seedling 3cm, culture bottle cap is opened, it is indoor
Then root culture medium is cleaned in hardening culture 5 days, move into the hole tray using 64 holes containing the peat soil after disinfection, it is big to be placed in plastics
Canopy keeps 85% or more humidity.
Embodiment 2
A kind of quick breeding method for tissue culture of purple brussels sprout, includes the following steps：
(1) explant is chosen and is sterilized：
1. choosing the tender stem apex of brussels sprout of robust growth as explant；
2. removing extra blade, stem apex is washed with neutral detergent, to remove the dirt on branch surface, is then used clean
Paper handkerchief blots surface moisture；
3. on superclean bench, first 10-30s is impregnated with 65% alcohol, then with aseptic water washing 3-5 times, each 3-
5min；
5-7min is sterilized 4. being transferred to immediately in 0.1% mercuric chloride solution, finally with sterile water washing 3-5 times, each 3-
5min, be sterilized explant.
(2) preparation of modified MS medium：
The formula of modified MS medium is：With Ca (NO3)2·4H2O replaces the CaCl in original MS culture mediums2·2H2O simultaneously adds
Add riboflavin, other components unchanged；Wherein Ca (NO3)2·4H2A concentration of 50mg/L of O, a concentration of 5mg/ of riboflavin
L。
(3) adventitious bud inducing：
The disinfection explant that step (1) is obtained, cuts away the lower end of stem apex, is then inoculated in the training for adventitious bud inducing
Base is supported, carries out Fiber differentiation 18 days, obtains brussels sprout adventitious bud；Inducing culturing condition is as follows：Temperature is 26 ± 3 DEG C, humidity
It is 50%, light application time is 12 hours/day, intensity of illumination 800-10001x；
The formula of culture medium for adventitious bud inducing is：In the modified MS medium add 6-BA, KT and
IAA；A concentration of 0.1mg/L of a concentration of 0.8mg/L of a concentration of 0.5mg/L of wherein 6-BA, KT, IAA.
(4) shoot proliferation：
Culture medium of the brussels sprout adventitious bud access that step (3) is obtained for shoot proliferation, carries out squamous subculture 20
It, obtains brussels sprout aseptic seedling；Shoot proliferation condition of culture is as follows：Temperature is 26 ± 3 DEG C, humidity 50%, light application time
For 12 hours/day, intensity of illumination 800-10001x；
The formula of culture medium for shoot proliferation is：On the basis of the modified MS medium add 6-BA, KT with
And a concentration of 0.05mg/L of a concentration of 0.5mg/L of a concentration of 0.5mg/L of IAA, wherein 6-BA, KT, IAA.
(5) sterile rootage：
The brussels sprout aseptic seedling that step (4) obtains cut numerous, and is kept for every plant there are one adventitious bud, access is used
Culture of rootage is carried out in the culture medium of sterile rootage 15 days, obtain brussels sprout seedling；Culture of rootage condition is as follows：Culture temperature
Degree is 26 ± 3 DEG C, humidity 50%, and light application time is 12 hours/day, intensity of illumination 800-10001x；
The formula of culture medium for sterile rootage is：IBA and IAA is added on the basis of the modified MS medium,
A concentration of 0.5mg/L of a concentration of 1mg/L of wherein IBA, IAA.
(6) hardening and transplanting：
In step (5) after brussels sprout growth of seedling to root long 0.5cm, height of seedling 3cm, culture bottle cap is opened, it is indoor
Then root culture medium is cleaned in hardening culture 5 days, move into the hole tray using 64 holes containing the peat soil after disinfection, it is big to be placed in plastics
Canopy keeps 85% or more humidity.
Embodiment 3
A kind of quick breeding method for tissue culture of green brussels sprout, includes the following steps：
(1) explant is chosen and is sterilized：
1. choosing the tender branch of brussels sprout of robust growth as explant；
2. washing branch with neutral detergent, to remove the dirt on branch surface, extra blade is then removed, totally
Paper handkerchief blots surface moisture；
3. on superclean bench, first 10-30s is impregnated with 70% alcohol, then with aseptic water washing 3-5 times, each 3-
5min；
8-12min is sterilized 4. being transferred to immediately in 0.3% mercuric chloride solution, finally with sterile water washing 3-5 times, each 3-
5min, be sterilized explant.
(2) preparation of modified MS medium：
The formula of modified MS medium is：With Ca (NO3)2·4H2O replaces the CaCl in original MS culture mediums2·2H2O simultaneously adds
Add riboflavin, other components unchanged；Wherein Ca (NO3)2·4H2A concentration of 70mg/L of O, a concentration of 5mg/ of riboflavin
L。
(3) adventitious bud inducing：
The disinfection explant that step (1) obtains is cut into the simple bud of 0.4-0.5cm long or double leaf stem sections, is then inoculated in
For the culture medium of adventitious bud inducing, Fiber differentiation is carried out 22 days, obtain brussels sprout adventitious bud；Inducing culturing condition is as follows：
Temperature is 26 ± 3 DEG C, humidity 80%, and light application time is 15 hours/day, intensity of illumination 1000-12001x；
The formula of culture medium for adventitious bud inducing is：In the modified MS medium add 6-BA, KT and
IAA；A concentration of 0.5mg/L of a concentration of 0.5mg/L of a concentration of 0.8mg/L of wherein 6-BA, KT, IAA.
(4) shoot proliferation：
Culture medium of the brussels sprout adventitious bud access that step (3) is obtained for shoot proliferation, carries out squamous subculture 20-
25 days, obtain brussels sprout aseptic seedling；Shoot proliferation condition of culture is as follows：Temperature is 26 ± 3 DEG C, humidity 65%, when illumination
Between be 15 hours/day, intensity of illumination 800-12001x；
The formula of culture medium for shoot proliferation is：On the basis of the modified MS medium add 6-BA, KT with
And a concentration of 5mg/L of a concentration of 0.1mg/L of a concentration of 2mg/L of IAA, wherein 6-BA, KT, IAA.
(5) sterile rootage：
The brussels sprout aseptic seedling that step (4) obtains cut numerous, and is kept for every plant there are one adventitious bud, access is used
Culture of rootage is carried out in the culture medium of sterile rootage 20 days, obtain brussels sprout seedling；Culture of rootage condition is as follows：Culture temperature
Degree is 26 ± 3 DEG C, humidity 65%, and light application time is 15 hours/day, intensity of illumination 1000-12001x；
The formula of culture medium for sterile rootage is：IBA and IAA is added on the basis of the modified MS medium,
A concentration of 5mg/L of a concentration of 1mg/L of wherein IBA, IAA.
(6) hardening and transplanting：
In step (5) after brussels sprout growth of seedling to root long 0.5cm, height of seedling 3cm, culture bottle cap is opened, it is indoor
Then root culture medium is cleaned in hardening culture 5 days, move into the hole tray using 64 holes containing the peat soil after disinfection, it is big to be placed in plastics
Canopy keeps 85% or more humidity.
Embodiment 4
A kind of quick breeding method for tissue culture of green brussels sprout, includes the following steps：
(1) explant is chosen and is sterilized：
1. choosing the tender stem apex of brussels sprout of robust growth as explant；
2. removing extra blade, stem apex is washed with neutral detergent, to remove the dirt on branch surface, is then used clean
Paper handkerchief blots surface moisture；
3. on superclean bench, first 10-30s is impregnated with 65% alcohol, then with aseptic water washing 3-5 times, each 3-
5min；
5-6min is sterilized 4. being transferred to immediately in 0.1% mercuric chloride solution, finally with sterile water washing 3-5 times, each 3-
5min, be sterilized explant.
(2) preparation of modified MS medium：
The formula of modified MS medium is：With Ca (NO3)2·4H2O replaces the CaCl in original MS culture mediums2·2H2O simultaneously adds
Add riboflavin, other components unchanged；Wherein Ca (NO3)2·4H2A concentration of 100mg/L of O, riboflavin it is a concentration of
0.3mg/L。
(3) adventitious bud inducing：
The disinfection explant that step (1) is obtained, cuts away the lower end of stem apex, is then inoculated in the training for adventitious bud inducing
Base is supported, carries out Fiber differentiation 18 days, obtains brussels sprout adventitious bud；Inducing culturing condition is as follows：Temperature is 26 ± 3 DEG C, humidity
It is 50%, light application time is 12 hours/day, intensity of illumination 800-10001x；
The formula of culture medium for adventitious bud inducing is：In the modified MS medium add 6-BA, KT and
IAA；A concentration of 0.05mg/L of a concentration of 1mg/L of a concentration of 0.6mg/L of wherein 6-BA, KT, IAA.
(4) shoot proliferation：
Culture medium of the brussels sprout adventitious bud access that step (3) is obtained for shoot proliferation, carries out squamous subculture 20
It, obtains brussels sprout aseptic seedling；Shoot proliferation condition of culture is as follows：Temperature is 26 ± 3 DEG C, humidity 50%, light application time
For 14 hours/day, intensity of illumination 800-10001x；
The formula of culture medium for shoot proliferation is：On the basis of the modified MS medium add 6-BA, KT with
And a concentration of 2.4mg/L of a concentration of 2mg/L of a concentration of 0.5mg/L of IAA, wherein 6-BA, KT, IAA.
(5) sterile rootage：
The brussels sprout aseptic seedling that step (4) obtains cut numerous, and is kept for every plant there are one adventitious bud, access is used
Culture of rootage is carried out in the culture medium of sterile rootage 15 days, obtain brussels sprout seedling；Culture of rootage condition is as follows：Culture temperature
Degree is 26 ± 3 DEG C, humidity 50%, and light application time is 12 hours/day, intensity of illumination 800-10001x；
The formula of culture medium for sterile rootage is：IBA and IAA is added on the basis of the modified MS medium,
A concentration of 4.5mg/L of a concentration of 0.5mg/L of wherein IBA, IAA.
(6) hardening and transplanting：
In step (5) after brussels sprout growth of seedling to root long 0.5cm, height of seedling 3cm, culture bottle cap is opened, it is indoor
Then root culture medium is cleaned in hardening culture 5 days, move into the hole tray using 64 holes containing the peat soil after disinfection, it is big to be placed in plastics
Canopy keeps 85% or more humidity.
Finally, it should be noted that the above embodiments are merely illustrative of the technical solutions of the present invention rather than is protected to the present invention
The limitation of range is protected, although being explained in detail to the present invention with reference to preferred embodiment, those skilled in the art should
Understand, technical scheme of the present invention can be modified or replaced equivalently, without departing from the essence of technical solution of the present invention
And range.
Claims (5)
1. a kind of quick breeding method for tissue culture of brussels sprout, includes the following steps：(1) explant is chosen and is sterilized, (2)
The preparation of culture medium, (3) adventitious bud inducing, (4) shoot proliferation, (5) sterile rootage and (6) hardening exist with transplanting, feature
In the preparation method of the culture medium is as follows：
(1) preparation of modified MS medium, is formulated and is：With Ca (NO3)2·4H2O replaces the CaCl in original MS culture mediums2·2H2O
And riboflavin is added, other components are unchanged；Ca (the NO in modified MS medium3)2·4H2A concentration of 50-100mg/ of O
L, a concentration of 0.3-5mg/L of riboflavin；
(2) culture medium for adventitious bud inducing is prepared, is formulated and is：In the modified MS medium add 6-BA, KT and
IAA；A concentration of 0.5-0.8mg/L of 6-BA in adventitious bud induction culture base, a concentration of 0.5-1mg/L of KT, the concentration of IAA
For 0.05-0.5mg/L；
(3) culture medium for shoot proliferation is prepared, is formulated and is：6-BA, KT are added on the basis of the modified MS medium
And IAA, a concentration of 0.5-2mg/L of 6-BA in subculture multiplication medium, a concentration of 0.1-2mg/L of KT, the concentration of IAA
For 0.05-5mg/L；
(4) culture medium for sterile rootage is prepared, is formulated and is：On the basis of the modified MS medium add IBA and
IAA, a concentration of 0.1-1mg/L of IBA, a concentration of 0.5-5mg/L of IAA in root media.
2. the quick breeding method for tissue culture of brussels sprout as described in claim 1, which is characterized in that in improvement MS cultures
Ca (NO in base3)2·4H2A concentration of 60mg/L of O, a concentration of 2.5mg/L of riboflavin.
3. the quick breeding method for tissue culture of brussels sprout as described in claim 1, which is characterized in that described for not
A concentration of 1mg/L, a concentration of 0.2mg/L of a concentration of 0.5mg/L of KT, IAA of 6-BA in the culture medium of normal bud induction.
4. the quick breeding method for tissue culture of brussels sprout as described in claim 1, which is characterized in that it is described for after
A concentration of 2mg/L, a concentration of 0.5mg/L of KT of 6-BA in the culture medium of generation proliferation.
5. the quick breeding method for tissue culture of brussels sprout as described in claim 1, which is characterized in that be used for nothing described
A concentration of 0.5mg/L of IBA in the culture medium that bacterium takes root, a concentration of 1mg/L of IAA.
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