CN105165614A - Brucea javanica seedling culture method based on tissue culture - Google Patents
Brucea javanica seedling culture method based on tissue culture Download PDFInfo
- Publication number
- CN105165614A CN105165614A CN201510620594.0A CN201510620594A CN105165614A CN 105165614 A CN105165614 A CN 105165614A CN 201510620594 A CN201510620594 A CN 201510620594A CN 105165614 A CN105165614 A CN 105165614A
- Authority
- CN
- China
- Prior art keywords
- seedling
- medium
- seed
- culture
- induction
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Cultivation Of Plants (AREA)
Abstract
The invention discloses a brucea javanica seedling culture method based on tissue culture and belongs to the technical field of Chinese medicinal herb breeding. The method specifically comprises the steps of seed collection, aseptic seedling culture, medium induction, multiple-shoot induction, multiple-shoot subculture multiplication, root induction and test-tube plantlet transplantation. By the adoption of the method, the problems of the prior art that time for artificial breeding of brucea javanica is long, yield is low, and large-area development can not be achieved are effectively solved. The method has the advantages that materials can be obtained conveniently, multiple-shoot induction rate can reach 65-80%, seedling development rate is over 85%, a large number of high-quality tissue culture seedlings can be bred in a short period of time so as to meet the requirement for mass artificial culture, growth is fast after transplantation and field planting, yield is increased by 30-35% compared with seed seedling transplantation, and the effective medicinal components of obtained brucea javanica are identical with those of wild brucea javanica.
Description
Technical field
The present invention relates to Chinese medicinal herbs raising technology, be specifically related to a kind of method of Java brucea nursery, especially the method for Java brucea tissue cultivating and seedling, belong to Chinese medicinal herbs raising technology field.
Background technology
Java brucea is quassia, formal name used at school: Bruceajavanica, another name: duck's egg, crow courage, crow egg-shaped thing, khosem, little China tree etc., Chang Wei great shrub or dungarunga, plant height 1 ~ 3 meter, complete stool is by yellow pubescence, bark has bitter taste, imparipinnate leaf alternate, medicinal part is dry mature fruit, and its root, Ye Yike are used as medicine.
Fruit of khosam is mainly containing alkaloid, glucoside and Flavonoid substances; Its seed is containing the bitter principle of the multiple guassins such as Brusatol.The kind benevolence of Java brucea is containing the fat oil such as oleic acid, linoleic acid, their content is up to 56.23%, wherein oleic acid is anticancer effective active composition, the brucea fruit oil of gained has certain antitumaous effect through clinical verification for many years, make 10% Emulsion of Seed Oils From Brucea Javanica clinically and be used for the treatment of digestive system tumor as cancer of the stomach, the cancer of the esophagus etc., particularly along with the development of interventional radiology, bruceolic oil emulsion has larger value to treatment primary carcinoma of liver aspect, and conventional bruceolic oil emulsion, mitomycin, lipiodol get involved Be very effective.Java brucea is cold in nature, bitter, slightly poisonous; Have clearing heat and detoxicating, desinsection, only dysentery, corrosion sting wart function, are used for the treatment of amoebic dysentery, malaria, the early stage disease such as blood fluke, bacillary dysentery, external curing corn, skin wart etc.In addition, Java brucea is utilized can also to manufacture some daily chemicals, as medicated soap, bathing lotion, hair-cream, hair oil, skin cream, agricultural missible oil, Java brucea cooling bath also can be used as mosquitocide.
Raising along with people's living standard and the demand to health, the purposes of Java brucea gets more and more, and people make again wild Java brucea endangered to the indiscriminate mining and serious waste of wild resource, and for satisfying the demand, people have to develop artificial planting technique.
In prior art, carry out Java brucea artificial cultivation, main employing planting seed seedling-cultivating method, this method has the defect of two aspects: one is that the cultivation time is long, yields poorly, and can not meet the demand of market to Java brucea; Two is that germination rate is low because Java brucea provenance is limited, large area can not develop Java brucea artificial planting, cannot form industrialization.
Summary of the invention
goal of the invention: the object of the invention is for the Java brucea artificial breeding time in prior art long, yield poorly, can not large area development problem, a kind of tissue cultivating and seedling method is provided, shortens Java brucea and breed the time, improve nursery planting percent, and Java brucea medicinal ingredient can be kept, after transplanting, growth is fast, large area can carry out Java brucea artificial cultivation, forms industrialization.
technical scheme: a kind of Java brucea tissue cultivating and seedling method of the present invention, specifically comprises the following steps:
1. collect seed: choosing is healthy and strong, do seed collecting maternal plant without the wild Java brucea plant of damage by disease and insect, after annual Frost's Descent solar term, seed maturity, can collect seed.
2. the cultivation of aseptic seedling, Java brucea seed is placed in 0.3% mercuric chloride solution soaking disinfection 15 ~ 18 minutes, superclean bench embathes 3 ~ 5 times with sterile water, embathes 5 ~ 8 minutes at every turn, then by planting seed in medium, cultivate under Indoor Natural light scattering.
3. inducing culture: take MS as minimal medium, add plant hormone and carry out Fiber differentiation, plant hormone is with benzyl purine (BA), concentration is 2.0 ~ 2.5mg/L, add the sucrose of 2.8% ~ 3% and the agar of 0.8% simultaneously, medium temperature keeps 25 DEG C ~ 28 DEG C, regulates pH value to be 6.0 ~ 6.5.
4. inducing clumping bud: after the aseptic seedling of step 2 cultivation grows to 5 ~ 6 centimetres, cut the top leaf stem section of top 2.5 ~ 3.0 centimeter length, namely access the medium culture of inducing has Multiple Buds to be formed after 28 ~ 30 days.
5. Multiple Buds shoot proliferation: inducing clumping bud to average up to 3.5 centimetres after (need 48 ~ 52 days), Multiple Buds is cut into 3 bud clusters, goes to the medium that another induce, after 25 days ~ 30 days, breed to 45 ~ 50 Multiple Buds for every bunch.
6. root induction: when the Multiple Buds of shoot proliferation has 70% to grow to 3 ~ 4 centimetres, cut by the bud list of stalwartness, and root induction on the medium forwarding new induction to, after 18 days, base portion otch starts to differentiate root.
7. the transplanting of test-tube plantlet: will physically well develop, has 2 ~ 3 test-tube plantlet bottle caps launching blade and opens, be placed in room temperature lower refining seedling and take out seedling after 3 ~ 5 days, the medium of root is washed away with clear water, in the booth having 65% ~ 75% degree of shading, be transplanted in seedbed or nutrition cup, water permeable, and with covered rearing with plastic film, open film every day to take a breath 45 minutes, atomized water spray keeps air humidity after more than 85%, 10 ~ 12 days, throw off film, can land for growing field crops be transplanted.
beneficial effect: compared with prior art, its beneficial effect is in the present invention:
Java brucea of the present invention organizes seedling-cultivating method, draw materials conveniently, inducing clumping bud rate reaches 65% ~ 80%, seedling reaches more than 85%, can breed a large amount of high-quality plantlet in vitro at short notice, to meet the needs of artificial a large amount of cultivation, after transplant planting, rapidly, output transplants increase by 30% ~ 35% than seed seedling-raising, and consistent with effective medicinal ingredient of wild Java brucea in growth.
Embodiment
Below in conjunction with concrete example of implementing, technical solution of the present invention is described in detail.
embodiment 1: a kind of Java brucea tissue cultivating and seedling, concrete steps are as follows:
1. collect seed: choosing is healthy and strong, do seed collecting maternal plant without the wild Java brucea plant of damage by disease and insect, after annual Frost's Descent solar term, seed maturity, can collect seed.
2. the cultivation of aseptic seedling, is placed in 0.3% mercuric chloride solution soaking disinfection 15 minutes by Java brucea seed, superclean bench embathes 3 times with sterile water, embathes 8 minutes at every turn, then by planting seed in medium, cultivate under Indoor Natural light scattering.
3. inducing culture: be minimal medium with MS, adds plant hormone and carries out Fiber differentiation, and plant hormone is with benzyl purine (BA), concentration is 2.0mg/L, add the sucrose of 2.8% and the agar of 0.8%, medium temperature keeps 25 DEG C simultaneously, regulates pH value to be 6.0.
4. inducing clumping bud: after the aseptic seedling of step 2 cultivation grows to 5 centimetres, cut the top leaf stem section of top 2.5 centimeter length, namely access the medium culture of inducing has Multiple Buds to be formed after 28 days.
5. Multiple Buds shoot proliferation: inducing clumping bud to average up to 3.5 centimetres after, Multiple Buds is cut into 3 bud clusters, goes to the medium that another induce, after 25 days, every bunch breed to 45 Multiple Buds.
6. root induction: when the Multiple Buds of shoot proliferation has 70% to grow to 3 centimetres, cut by the bud list of stalwartness, and root induction on the medium forwarding new induction to, after 18 days, base portion otch starts to differentiate root.
7. the transplanting of test-tube plantlet: will physically well develop, has 2 test-tube plantlet bottle caps launching blade and opens, be placed in room temperature lower refining seedling and take out seedling after 3 days, the medium of root is washed away with clear water, in the booth having 65% degree of shading, be transplanted in seedbed or nutrition cup, water permeable, and with covered rearing with plastic film, open film every day to take a breath 45 minutes, atomized water spray keeps air humidity after more than 85%, 10 days, throw off film, can land for growing field crops be transplanted.
embodiment 2: a kind of Java brucea tissue cultivating and seedling, comprises following concrete steps:
1. collect seed: choosing is healthy and strong, do seed collecting maternal plant without the wild Java brucea plant of damage by disease and insect, after annual Frost's Descent solar term, seed maturity, can collect seed.
2. the cultivation of aseptic seedling, is placed in 0.3% mercuric chloride solution soaking disinfection 16 minutes by Java brucea seed, superclean bench embathes 4 times with sterile water, embathes 6 minutes at every turn, then by planting seed in medium, cultivate under Indoor Natural light scattering.
3. inducing culture: be minimal medium with MS, adds plant hormone and carries out Fiber differentiation, and plant hormone is with benzyl purine (BA), concentration is 2.2mg/L, add the sucrose of 2.9% and the agar of 0.8%, medium temperature keeps 26 DEG C simultaneously, regulates pH value to be 6.3.
4. inducing clumping bud: after the aseptic seedling of step 2 cultivation grows to 5.5 centimetres, cut the top leaf stem section of top 2.8 centimeter length, namely access the medium culture of inducing has Multiple Buds to be formed after 29 days.
5. Multiple Buds shoot proliferation: inducing clumping bud to average up to 3.5 centimetres after (need 48 ~ 52 days), Multiple Buds is cut into 3 bud clusters, goes to the medium that another induce, after 28 days, breed to 45 ~ 50 Multiple Buds for every bunch.
6. root induction: when the Multiple Buds of shoot proliferation has 70% to grow to 3.5 centimetres, cut by the bud list of stalwartness, and root induction on the medium forwarding new induction to, after 20 days, base portion otch starts to differentiate root.
7. the transplanting of test-tube plantlet: will physically well develop, has 2 test-tube plantlet bottle caps launching blade and opens, be placed in room temperature lower refining seedling and take out seedling after 4 days, the medium of root is washed away with clear water, in the booth having 70% degree of shading, be transplanted in seedbed or nutrition cup, water permeable, and with covered rearing with plastic film, open film every day to take a breath 45 minutes, atomized water spray keeps air humidity after more than 85%, 11 days, throw off film, can land for growing field crops be transplanted.
embodiment 3: a kind of Java brucea tissue cultivating and seedling, its concrete steps are as follows:
1. collect seed: choosing is healthy and strong, do seed collecting maternal plant without the wild Java brucea plant of damage by disease and insect, after annual Frost's Descent solar term, seed maturity, can collect seed.
2. the cultivation of aseptic seedling, is placed in 0.3% mercuric chloride solution soaking disinfection 18 minutes by Java brucea seed, superclean bench embathes 5 times with sterile water, embathes 5 minutes at every turn, then by planting seed in medium, cultivate under Indoor Natural light scattering.
3. inducing culture: be minimal medium with MS, add plant hormone and carry out Fiber differentiation, plant hormone is 2.5mg/L by benzyl purine (BA), concentration, and add the sucrose of 3% and the agar of 0.8%, medium temperature keeps 28 DEG C simultaneously, regulates pH value to be 6.5.
4. inducing clumping bud: after the aseptic seedling of step 2 cultivation grows to 6 centimetres, cut the top leaf stem section of top 3.0 centimeter length, namely access the medium culture of inducing has Multiple Buds to be formed after 30 days.
5. Multiple Buds shoot proliferation: inducing clumping bud to average up to 3.5 centimetres after (need 48 ~ 52 days), Multiple Buds is cut into 3 bud clusters, goes to the medium that another induce, after 30 days, breed to 50 Multiple Buds for every bunch.
6. root induction: when the Multiple Buds of shoot proliferation has 70% to grow to 4 centimetres, cut by the bud list of stalwartness, and root induction on the medium forwarding new induction to, after 20 days, base portion otch starts to differentiate root.
7. the transplanting of test-tube plantlet: will physically well develop, has 3 test-tube plantlet bottle caps launching blade and opens, be placed in room temperature lower refining seedling and take out seedling after 5 days, the medium of root is washed away with clear water, in the booth having 75% degree of shading, be transplanted in seedbed or nutrition cup, water permeable, and with covered rearing with plastic film, open film every day to take a breath 45 minutes, atomized water spray keeps air humidity after more than 85%, 12 days, throw off film, can land for growing field crops be transplanted.
Claims (2)
1. a Java brucea tissue cultivating and seedling method, is characterized in that comprising the following steps:
(1) collect seed: choosing is healthy and strong, do seed collecting maternal plant without the wild Java brucea plant of damage by disease and insect, after annual Frost's Descent solar term, seed maturity, can collect seed;
(2) cultivation of aseptic seedling, Java brucea seed is placed in 0.3% mercuric chloride solution soaking disinfection 15 ~ 18 minutes, superclean bench embathes 3 ~ 5 times with sterile water, embathes 5 ~ 8 minutes at every turn, then by planting seed in medium, cultivate under Indoor Natural light scattering;
(3) inducing culture: take MS as minimal medium, add plant hormone and carry out Fiber differentiation, plant hormone is with benzyl purine (BA), concentration is 2.0 ~ 2.5mg/L, add the sucrose of 2.8% ~ 3% and the agar of 0.8% simultaneously, medium temperature keeps 25 DEG C ~ 28 DEG C, regulates pH value to be 6.0 ~ 6.5;
(4) inducing clumping bud: after the aseptic seedling of cultivating when step (2) grows to 5 ~ 6 centimetres, cut the top leaf stem section of top 2.5 ~ 3.0 centimeter length, accesses the medium culture Multiple Buds of inducing;
(5) Multiple Buds shoot proliferation: inducing clumping bud to average up to 3.5 centimetres after, Multiple Buds is cut into 3 bud clusters, goes to the medium that another was induced, allow adventitious buds proliferation;
(6) root induction: when the Multiple Buds of shoot proliferation has 70% to grow to 3 ~ 4 centimetres, the bud list of stalwartness is cut, and root induction on the medium forwarding new induction to.
2. Java brucea tissue cultivating and seedling method according to claim 1, characterized by further comprising the transplanting of step (7) test-tube plantlet: after inducing clumping bud is taken root, to physically well develop, there are 2 ~ 3 test-tube plantlet bottle caps launching blade open, be placed in room temperature lower refining seedling and take out seedling after 3 ~ 5 days, the medium of root is washed away with clear water, in the booth having 65% ~ 75% degree of shading, be transplanted in seedbed or nutrition cup, water permeable, and with covered rearing with plastic film, open film every day to take a breath 45 minutes, atomized water spray keeps air humidity more than 85%, after 10 ~ 12 days, throw off film, land for growing field crops can be transplanted.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510620594.0A CN105165614A (en) | 2015-09-27 | 2015-09-27 | Brucea javanica seedling culture method based on tissue culture |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510620594.0A CN105165614A (en) | 2015-09-27 | 2015-09-27 | Brucea javanica seedling culture method based on tissue culture |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105165614A true CN105165614A (en) | 2015-12-23 |
Family
ID=54888502
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510620594.0A Pending CN105165614A (en) | 2015-09-27 | 2015-09-27 | Brucea javanica seedling culture method based on tissue culture |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105165614A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107853177A (en) * | 2017-11-14 | 2018-03-30 | 广西壮族自治区药用植物园 | The method of Java brucea tissue-culturing quick-propagation |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102972194A (en) * | 2012-12-14 | 2013-03-20 | 茂名市天一农业科技发展有限公司 | Manual cultivation method of khosam |
| CN104206278A (en) * | 2014-09-12 | 2014-12-17 | 南京通泽农业科技有限公司 | Rapid propagation method for cultivating brucea javanica suspension cells |
| CN104904547A (en) * | 2015-06-06 | 2015-09-16 | 黄振忠 | Nutrition cup seedling raising method for brucea fruits |
-
2015
- 2015-09-27 CN CN201510620594.0A patent/CN105165614A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102972194A (en) * | 2012-12-14 | 2013-03-20 | 茂名市天一农业科技发展有限公司 | Manual cultivation method of khosam |
| CN104206278A (en) * | 2014-09-12 | 2014-12-17 | 南京通泽农业科技有限公司 | Rapid propagation method for cultivating brucea javanica suspension cells |
| CN104904547A (en) * | 2015-06-06 | 2015-09-16 | 黄振忠 | Nutrition cup seedling raising method for brucea fruits |
Non-Patent Citations (2)
| Title |
|---|
| ME.WAGIH ET.AL.,: "Improved production of the indole alkaloid canthin-6-one from cell suspension culture of Brucea javanica (L.) Merr.", 《INDIAN JOURNAL OF SCIENCE AND TECHNOLOGY》 * |
| 夏静等: "鸦胆子茎节组培快繁体系的建立", 《中国现代中药》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107853177A (en) * | 2017-11-14 | 2018-03-30 | 广西壮族自治区药用植物园 | The method of Java brucea tissue-culturing quick-propagation |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103141260B (en) | Method for ecologically interplanting sarcandra glabra under phyllostachys edulis forest | |
| Dharani et al. | Common antimalarial trees and shrubs of East Africa | |
| CN102440165A (en) | Method of cultivating amygdalus persica A.N. trees | |
| KR101466474B1 (en) | The culturing method of medicinal herbs | |
| CN103283428A (en) | Greenhouse artificial cultivation method of dendrobium | |
| CN104429497A (en) | Planting method for lilies | |
| CN106688893A (en) | Beautiful millettia root tissue culture seedling raising method | |
| CN105284314A (en) | Cultivation method for passion fruit virus-free seedling | |
| CN104041417A (en) | Morinda officinalis tissue culture breeding method | |
| CN105684844A (en) | Dendrobium officinale planting method | |
| CN109258460A (en) | Micro-stem tip culture combines the breeding method of heat treatment acquisition Zengcheng honey chrysanthemum detoxic seedling | |
| CN104904547A (en) | Nutrition cup seedling raising method for brucea fruits | |
| CN104160849A (en) | Raspberry planting method | |
| CN103503776A (en) | Radix sophorae tonkinensis tissue culture seedling raising method | |
| CN103891495A (en) | Multivariate space ecological blueberry planting pattern | |
| CN104663176A (en) | Planting method for honeysuckle | |
| CN105393900A (en) | Built-on-stilt seedbed standardized organic planting method under dendrobium huoshanense forest | |
| CN103229642A (en) | Breeding method for improved variety of Lonicera japonica Thunb No.1 | |
| CN107548921A (en) | The cultural method of caper in a kind of landscape greenhouse | |
| CN104813938A (en) | Panax notoginseng tissue culture seedling raising method | |
| CN105875166A (en) | Amomum tsao-ko biomimetic seedling culturing method | |
| CN103229716A (en) | Method for seed selection of excellent honeysuckle flower Lonicera japonica Thunb II variety | |
| CN112655482A (en) | High-efficiency high-yield cultivation method for interplanting dragon fruits and pineapples in greenhouse | |
| CN107360934A (en) | A kind of method for planting fig fruit tree | |
| CN106258372A (en) | A kind of implantation methods of Caulis et folium pavettae hongkongensis Passifolra edulis |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20151223 |
|
| WD01 | Invention patent application deemed withdrawn after publication |