CN104885942B - Chemical disinfection tissue culture method for rhododendron simsii planch - Google Patents
Chemical disinfection tissue culture method for rhododendron simsii planch Download PDFInfo
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Abstract
The invention relates to a chemical disinfection tissue culture method for rhododendron simsii planch. The chemical disinfection tissue culture method for rhododendron simsii planch comprises the steps of culture vessel disinfection, culture medium preparation, induction culture, enrichment culture, rooting culture and bottle seedling transplanting. According to the chemical disinfection tissue culture method for rhododendron simsii planch, the purpose of disinfection or bacteriostasis can be achieved by a chemical reagent; in the culture medium preparation process, a culture vessel and a culture medium do not need to be disinfected at high temperature and high pressure, the workload and the energy consumption can be reduced, and the rhododendron simsii planch tissue culture link can be simplified; the operation is simple, and the preparation is implemented according to different culture medium formulae; the practicability is high, the popularization is good, the tissue culture cost of rhododendron simsii planch can be effectively lowered, and the cost can be generally reduced by no less than 10%.
Description
Technical field
The present invention relates to a kind of method for plant tissue culture, be specifically related to a kind of Folium Rhododendri Simsii chemical disinfection tissue culture method, belong to biological skill
Art field.
Background technology
Rhododendron (RhododendronLinn.) plant is world-renowned ornamental plant, is one of big famous flower of China ten, is also
World-renowned ornamental flower. its gorgeous colorful pattern and proper potted flower moulding are always in occupation of China's flowers in Spring Festival fast sale kind
One of.Folium Rhododendri Simsii generally uses seed, cuttage, press strip and engrafting method breeding, but by kind or kind, season, maternal plant material
Limit etc. factor, it is difficult to carry out substantial amounts of commercially producing.Ghent azalea it has been successfully set up first from Anderon in 1975
The culture in vitro system of (Rhododendron xx), tissue culture has become the important means of Folium Rhododendri Simsii breeding, is widely used in planting
In the commercially producing of matter protection of resources, genetic breeding and clone nursery stock.For specific Folium Rhododendri Simsii kind, kind, its from
The selection that it is critical only that minimal medium of body reproductive success, to added hormone kind in the induction of implant outside and enrichment culture
And the optimization of proportioning, and to the regulation and control of envirment factor in rooting culture.But, due to Folium Rhododendri Simsii kind and interracial difference,
In tissue culture procedures, the foundation of the rooting culture system of subculture enrichment culture, root induction, tissue cultured seedling, is still tissue culture
The problem that middle urgent problem, particularly Multiple Buds and Differentiation ration of adventitious buds are low, is the bottleneck of tissue-culturing rapid propagation.
Cuculus polioephalus tissue culture, on the one hand can expand its breeding coefficient, thus realize biological control；On the other hand, Ke Yi
On the basis of preserving its original merit, it is achieved the Germ-plasma resources protection of Cuculus polioephalus and transgenic research etc..Cuculus polioephalus tissue culture
Cost mainly include tissue cultured seedling produce needed for culture medium, facilities and equipment, power supply cost, consumptive material and cost of labor.Conventional
The outer implant of Cuculus polioephalus method for tissue culture requirement is seeded in ability normal growth in the aseptic culture medium after autoclave sterilization, and power consumption is big,
Manual operation cost is high.If culture medium can be transformed by the method for chemical disinfection, culture medium is made to be made without high temperature height
Pressure sterilizing, it becomes possible to obtain the normal growth of Cuculus polioephalus.So, on the one hand can reduce Cuculus polioephalus tissue culture facilities and equipment is wanted
Asking, especially need not the pressure cooker equipment of the required configuration of autoclave sterilization, reduce cost of investment, power consumption also will significantly
Degree reduces；On the other hand, using this culture medium to carry out Cuculus polioephalus tissue culture, group training link is greatly simplified, manually-operated speed
Degree is greatly improved, thus reduces cost of labor.Additionally, due to improved culture medium self has sterilization, antibacterial or antibacterial
Effect, can effectively control the pollution problem in tissue culture procedures, will not produce Cuculus polioephalus growth again and poison or suppression, the most not shadow
Ring the normal growth of Cuculus polioephalus, radically simplify Cuculus polioephalus tissue culture.
Summary of the invention
It is an object of the invention to provide a kind of Folium Rhododendri Simsii chemical disinfection tissue culture method.
It is an object of the invention to be achieved through the following technical solutions.
A kind of Folium Rhododendri Simsii chemical disinfection tissue culture method of the present invention, including culture vessel sterilization, culture medium preparation, inducing culture,
Enrichment culture, root culture and bottle transplantation of seedlings, it is characterised in that:
1. culture vessel sterilization: the stainless steel disc of culture bottle bottle cap and inoculation is blue or green at 100mg/L sodium hypochlorite+2mg/L
The aqueous solution of mycin+100mg/L calcium propionate soaks no less than 10h, saves backup；
2. culture medium preparation: inducing culture is ER+6～15mg/L ZT+5～15mg/L IAA+40～100mg/L hypochlorous acid
Sodium+2～5mg/L penicillin+50～100mg/L calcium propionate+20g/L sucrose+3.0g/L agar powder, pH5.8；Enrichment culture
Base is: ER+1～3mg/L ZT+0.5～1.0mg/L IAA+40～100mg/L sodium hypochlorite+2～5mg/L penicillin
+ 50～100mg/L calcium propionate+20g/L sucrose+3.0g/L agar powders, pH5.8；Root media is 1/2ER+0.5～1.5mg/L
NAA+0.1～0.5IBA mg/L+40～100mg/L sodium hypochlorite+2～5mg/L penicillin+50～100mg/L calcium propionate
+ 20g/L sucrose+3.0g/L agar powder, pH5.8；Described ER culture medium is nineteen sixty-five, ER culture medium disclosed in Eriksson；
Described ZT is zeatin；Described NAA is-naphthalene acetic acid；Described IAA is heteroauxing；Described IBA is indolebutyric acid；
During preparation culture medium, first claim sucrose and agar powder in each culture medium prescription, add the tap water of culture medium cumulative volume 1/2, be heated to
Agar powder is completely dissolved, then after assorting other raw materials by each culture medium prescription, constant volume, then with NaOH or 1mol/L of 1mol/L
HCl adjust pH value to 5.8, be dispensed in the culture vessel sterilized, sealing, standby after cooled and solidified；
3. inducing culture: take Cuculus polioephalus terminal bud or the alabastrum of stalwartness, first tap water rinses surface dirt, peels off outer bract, use volume
Ratio is to soak 2～3s in 75% ethanol, then is 0.1%HgCl by weight ratio2Sterilization 6～10min, aseptic water washing 3～5 times；
On superclean bench, peel off the bract of 2～3 layers, excise base portion brownization part, be inoculated on inducing culture, shape after 40d
Become some budlets；Culturing room's temperature is 25～28 DEG C, illumination 12h, and intensity of illumination is 1200～1500lx；
4. enrichment culture: proceed in proliferated culture medium by the budlet of inducing culture, progressively induces Multiple Buds and develops into after 30d
Do not has the crowd shoots of root；Crowd shoots is cut into the stem section of band bud, is transferred in new proliferated culture medium, every 30d switching one
Secondary；Culturing room's temperature is 25～28 DEG C, illumination 12h, and intensity of illumination is 1200～1500lx；
5. root culture: when the crowd shoots of enrichment culture grows to 4～6cm, crowd shoots is cut into individual plant, is inoculated into life
In root culture medium, after 25d, form whole plant；Culturing room's temperature is 25～28 DEG C, illumination 12h, intensity of illumination be 1200～
1500lx；
6. bottle transplantation of seedlings: the whole plant obtained by root culture takes out, and cleans the culture medium of root, with the carbendazim of 800 times
After soaking 30min, transplanting to cultivation matrix, described cultivation matrix is made up of peat soil and perlite, and its weight ratio is 2: 1；
Booth upper strata hides with 75% shading net, and the ambient temperature of transplanting is 22～28 DEG C.
The application effect of the present invention:
Due to the chemical disinfection tissue culture method of the present invention, be to utilize chemical reagent to add in various culture medium, reach sterilizing or
Antibacterial effect, so, the culture medium of preparation is without autoclave sterilization.Therefore, in Folium Rhododendri Simsii inducing culture, propagation training
Foster, the impact in each stage of root culture, in addition to considering the evaluation index commonly used, the problem that it is also contemplated that pollution rate.
1. inducing culture: experiments prove that, the chemical disinfection tissue culture method of a kind of Folium Rhododendri Simsii of the present invention and conventional Folium Rhododendri Simsii group
Culture method is compared, and result is as shown in table 1, survival rate, pollution rate and the mortality rate that the outer implant in the inducing culture stage is cultivated
All there is no too big difference.
The impact on Folium Rhododendri Simsii inducing culture of the chemical disinfection tissue culture method of table 1 present invention
| Tissue culture mode | Average survival (%) | Average pollution rate (%) | Average mortality (%) |
| Conventional tissue culture method | 45 | 36 | 19 |
| Chemical disinfection tissue culture method | 50 | 33 | 17 |
2. enrichment culture: the chemical disinfection tissue culture method of the present invention trains proliferation times and the comparison of pollution rate of Folium Rhododendri Simsii with conventional group,
Result is as shown in table 2, and proliferation times and two indexs of pollution rate all do not have too big difference.Upgrowth situation at bottle Seedling is seen, this
The culture medium of the chemical disinfection tissue culture method of invention owing to without autoclave sterilization, hormone and nutrient loss are less,
Its Folium Rhododendri Simsii tissue cultured seedling is greener, more strong.
The chemical disinfection tissue culture method of table 2 present invention is on Folium Rhododendri Simsii proliferation times and the impact of pollution rate
| Tissue culture mode | Average proliferation multiple | Average pollution rate (%) | Upgrowth situation |
| Conventional tissue culture method | 5.6 | 0.5 | Seedling is light green, weak |
| Chemical disinfection tissue culture method | 6.0 | 0 | Miao Lv, strong |
3. the chemical disinfection tissue culture method of the present invention is on Folium Rhododendri Simsii rooting rate and the impact of pollution rate: in rooting process, the present invention
Chemical disinfection tissue culture method compared with conventional Folium Rhododendri Simsii tissue culture method, result is as shown in table 3, and rooting rate and pollution rate all do not have
There is too big difference；In terms of the upgrowth situation of bottle Seedling, by the method for the present invention, Folium Rhododendri Simsii tissue cultured seedling stem is thicker, and blade is bigger.
The Folium Rhododendri Simsii rooting situation of the chemical disinfection tissue culture method of table 3 present invention
4. cost analysis: the chemical disinfection tissue culture method of the present invention is shown in Table 4 with the cost analysis of conventional group training.
Of the present invention group of training eliminates autoclaving link, simplifies group training program, improves work efficiency.From can directly calculate
Expense calculate, can save every year about 50,000 yuan (with prepare every day 50L culture medium calculate).It addition, also have some not allow
The project easily calculated, as pressure cooker keeps in repair, safety, saving space etc.:
The Folium Rhododendri Simsii simplicity tissue culture method of table 4 present invention is cultivated and the cost analysis table of conventional group training
The invention have the advantages that and beneficial effect:
1., in the Folium Rhododendri Simsii chemical disinfection tissue culture method of the present invention, culture vessel and culture medium, all without autoclave sterilization, reduce
Workload and energy resource consumption, simplify Folium Rhododendri Simsii group training link, reduce Folium Rhododendri Simsii group and train cost.
2. the Folium Rhododendri Simsii chemical disinfection tissue culture method of the present invention, simple to operate, as long as after by different culture medium prescription preparations,
Practical, generalization is good.
3. the Folium Rhododendri Simsii chemical disinfection tissue culture method of the present invention compares with conventional Folium Rhododendri Simsii tissue culture method, can reduce cost 10%
Above.
Detailed description of the invention
In order to be further elucidated with the present invention rather than limit the present invention, it is illustrated below in conjunction with embodiment.
Embodiment one: a kind of Folium Rhododendri Simsii chemical disinfection tissue culture method
A kind of Folium Rhododendri Simsii chemical disinfection tissue culture method, comprises the following steps:
1. culture vessel sterilization: the stainless steel disc of culture bottle bottle cap and inoculation is blue or green at 100mg/L sodium hypochlorite+2mg/L
The aqueous solution of mycin+100mg/L calcium propionate soaks 10h, saves backup.
2. culture medium preparation: inducing culture is ER+10mg/L ZT+10mg/L IAA+80mg/L sodium hypochlorite+3mg/L
Penicillin+50mg/L calcium propionate+20g/L sucrose+3.0g/L agar powder, pH5.8；Proliferated culture medium is: ER+1mg/L ZT+1.0
Mg/L IAA+80mg/L sodium hypochlorite+2mg/L penicillin+50mg/L calcium propionate+20g/L sucrose+3.0g/L agar powder,
pH5.8；Root media is 1/2ER+1.0mg/L NAA+0.5IBA mg/L+80mg/L sodium hypochlorite+2mg/L penicillium sp
Element+80mg/L calcium propionate+20g/L sucrose+3.0g/L agar powder, pH5.8；During preparation culture medium, first claim each culture medium prescription
Middle sucrose and agar powder, add the tap water of culture medium cumulative volume 1/2, is heated to agar powder and is completely dissolved, then by each cultivation basigamy
After other raw materials of Fang Peiqi, constant volume, then with the HCl of NaOH or 1mol/L of 1mol/L adjust pH value to 5.8, subpackage
In the culture vessel sterilized, sealing, standby after cooled and solidified.
3. inducing culture: take Cuculus polioephalus alabastrum or the terminal bud of stalwartness, first tap water rinses surface dirt, peels off outer bract, use volume
Ratio is to soak 2～3s in 75% ethanol, then is 0.1%HgCl by weight ratio2Sterilization 8min, aseptic water washing 3 times.In ultra-clean work
In station, peel off the bract of 2～3 layers, excise base portion brownization part, be inoculated on inducing culture, can be formed a large amount of after 40d
Budlet.Culturing room's temperature is 25～28 DEG C, illumination 12h, and intensity of illumination is 1200～1500lx.
4. enrichment culture: proceed in proliferated culture medium by the budlet of inducing culture, progressively induces Multiple Buds and develops into after 30d
Do not has the crowd shoots of root.Crowd shoots is cut into the stem section of band bud, is transferred in new proliferated culture medium, every 30d switching one
Secondary, the effect bred in a large number can be reached.Culturing room's temperature is 25～28 DEG C, illumination 12h, and intensity of illumination is 1200～1500
lx。
5. root culture: when the crowd shoots of enrichment culture grows to 4～6cm, crowd shoots is cut into individual plant, is inoculated into life
In root culture medium, after 25d, form whole plant；Culturing room's temperature is 25～28 DEG C, illumination 12h, intensity of illumination be 1200～
1500lx。
6. bottle transplantation of seedlings: the whole plant obtained by root culture takes out, and cleans the culture medium of root, with the carbendazim of 800 times
Soaking and transplant after 30min to peat soil and substrate that perlite ratio is 2: 1, booth upper strata hides with 75% shading net,
The ambient temperature transplanted is 22～28 DEG C.
Claims (4)
1. a Folium Rhododendri Simsii chemical disinfection tissue culture method, including culture vessel sterilization, culture medium preparation, inducing culture, propagation
Cultivation, root culture and bottle transplantation of seedlings, it is characterised in that:
(1) culture vessel sterilization: by the stainless steel disc of culture bottle bottle cap and inoculation at 100mg/L sodium hypochlorite+2mg/L
The aqueous solution of penicillin+100mg/L calcium propionate soaks no less than 10h, saves backup；
(2) culture medium preparation: inducing culture is ER+6～15mg/L ZT+5～15mg/L IAA+40～100mg/L chlorine
Acid sodium+2～5mg/L penicillin+50～100mg/L calcium propionate+20g/L sucrose+3.0g/L agar powder, pH5.8；Propagation training
Foster base is: ER+1～3mg/L ZT+0.5～1.0mg/L IAA+40～100mg/L sodium hypochlorite+2～5mg/L penicillin
+ 50～100mg/L calcium propionate+20g/L sucrose+3.0g/L agar powders, pH5.8；Root media is 1/2ER+0.5～1.5mg/L
NAA+0.1～0.5IBA mg/L+40～100mg/L sodium hypochlorite+2～5mg/L penicillin+50～100mg/L calcium propionate
+ 20g/L sucrose+3.0g/L agar powder, pH5.8；Described ER is nineteen sixty-five, ER culture medium disclosed in Eriksson；Described
ZT is zeatin；Described NAA is-naphthalene acetic acid；Described IAA is heteroauxing；Described IBA is indolebutyric acid；Preparation
During culture medium, first claim sucrose and agar powder in each culture medium prescription, add the tap water of culture medium cumulative volume 1/2, be heated to agar
Powder is completely dissolved, then after assorting other raw materials by each culture medium prescription, constant volume, then with NaOH or 1mol/L of 1mol/L
HCl adjust pH value to 5.8, be dispensed in the culture vessel sterilized, sealing, standby after cooled and solidified；
(3) inducing culture: take the Cuculus polioephalus alabastrum of stalwartness, first tap water rinses surface dirt, peels off outer bract, by volume ratio be
75% ethanol soaks 2～3s, then is 0.1%HgCl by weight ratio2Sterilization 6～10min, aseptic water washing 3～5 times；Ultra-clean
On workbench, peel off the bract of 2～3 layers, excise base portion brownization part, be inoculated on inducing culture, formed some after 40d
Budlet；Culturing room's temperature is 25～28 DEG C, illumination 12h, and intensity of illumination is 1200～1500lx；
(4) enrichment culture: proceed in proliferated culture medium by the budlet of inducing culture, progressively induces Multiple Buds and grows after 30d
Become not have the crowd shoots of root；Crowd shoots being cut into the stem section of band bud, is transferred in new proliferated culture medium, every 30d transfers
Once；Culturing room's temperature is 25～28 DEG C, illumination 12h, and intensity of illumination is 1200～1500lx；
(5) root culture: when the crowd shoots of enrichment culture grows to 4～6cm, crowd shoots is cut into individual plant, is inoculated into
On root media, after 25d, form whole plant；Culturing room's temperature is 25～28 DEG C, illumination 12h, intensity of illumination be 1200～
1500lx；
(6) bottle transplantation of seedlings: the whole plant obtained by root culture takes out, and cleans the culture medium of root, with many bacterium of 800 times
After 30min is soaked in spirit, transplanting to cultivation matrix, described cultivation matrix is made up of peat soil and perlite, and its weight ratio is 2:
1；Booth upper strata hides with 75% shading net, and the ambient temperature of transplanting is 22～28 DEG C.
A kind of Folium Rhododendri Simsii chemical disinfection tissue culture method the most according to claim 1, it is characterised in that described inducing culture is
ER+10mg/L ZT+10mg/L IAA+80mg/L sodium hypochlorite+3mg/L penicillin+50mg/L calcium propionate+20g/L sucrose
+ 3.0g/L agar powder, pH5.8.
A kind of Folium Rhododendri Simsii chemical disinfection tissue culture method the most according to claim 1, it is characterised in that described proliferated culture medium is:
ER+1mg/L ZT+1.0mg/L IAA+80mg/L sodium hypochlorite+2mg/L penicillin+50mg/L calcium propionate+20g/L sucrose
+ 3.0g/L agar powder, pH5.8.
A kind of Folium Rhododendri Simsii chemical disinfection tissue culture method the most according to claim 1, it is characterised in that described root media is
1/2ER+1.0mg/L NAA+0.5IBA mg/L+80mg/L sodium hypochlorite+2mg/L penicillin+80mg/L calcium propionate+20
G/L sucrose+3.0g/L agar powder, pH5.8.
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| CN105248281A (en) * | 2015-11-03 | 2016-01-20 | 钟煜 | Preparation method of raspberry induction culture medium |
| CN106613995A (en) * | 2017-01-18 | 2017-05-10 | 福建农林大学 | Tissue culture method for culturing anoectochilus roxburghii by bacteriostatic culture medium |
| CN114847162A (en) * | 2022-05-19 | 2022-08-05 | 福建农林大学 | Azalea in vitro culture and micro-bud grafting method |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101433185A (en) * | 2007-11-13 | 2009-05-20 | 刘磊 | Inducement substrate of regenerated plant from anther of berry |
| CN102668991A (en) * | 2012-06-14 | 2012-09-19 | 宁夏农垦西夏王实业有限公司 | Application of penicillin to simple test-tube breeding of grapes and novel technology for test-tube breeding of grapes |
| CN103952439A (en) * | 2014-05-04 | 2014-07-30 | 福建农林大学 | Method for carrying out bacteriostatic culture after genetic transformation of arabidopsis thaliana |
| CN104106466A (en) * | 2014-06-12 | 2014-10-22 | 南京工业大学大丰海洋产业研究院 | Explant disinfection method in plant tissue culture |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6140708A (en) * | 1984-07-31 | 1986-02-27 | ライオン株式会社 | Artificial seed |
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2015
- 2015-05-26 CN CN201510274041.4A patent/CN104885942B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101433185A (en) * | 2007-11-13 | 2009-05-20 | 刘磊 | Inducement substrate of regenerated plant from anther of berry |
| CN102668991A (en) * | 2012-06-14 | 2012-09-19 | 宁夏农垦西夏王实业有限公司 | Application of penicillin to simple test-tube breeding of grapes and novel technology for test-tube breeding of grapes |
| CN103952439A (en) * | 2014-05-04 | 2014-07-30 | 福建农林大学 | Method for carrying out bacteriostatic culture after genetic transformation of arabidopsis thaliana |
| CN104106466A (en) * | 2014-06-12 | 2014-10-22 | 南京工业大学大丰海洋产业研究院 | Explant disinfection method in plant tissue culture |
Non-Patent Citations (3)
| Title |
|---|
| Growth regulator effect on in vitro regeneration of rhododendron cultivars;H. Vejsadová;《Hort. Sci. (Prague)》;20081231(第2期);第90-94页 * |
| 杜鹃花组织培养技术研究;陈新平等;《江苏林业科技》;20031031;第30卷(第5期);第10-12页 * |
| 杜鹃花组织培养研究概况;郭秀莲等;《时珍国医国药》;20091231;第20卷(第9期);第2129-2131页 * |
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