CN102388807A - Rapid propagation method for tissue culture of wild Thymus Mongolicus Ronn and application thereof - Google Patents
Rapid propagation method for tissue culture of wild Thymus Mongolicus Ronn and application thereof Download PDFInfo
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- CN102388807A CN102388807A CN2011103324672A CN201110332467A CN102388807A CN 102388807 A CN102388807 A CN 102388807A CN 2011103324672 A CN2011103324672 A CN 2011103324672A CN 201110332467 A CN201110332467 A CN 201110332467A CN 102388807 A CN102388807 A CN 102388807A
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Abstract
The invention discloses a rapid propagation method for tissue culture of wild Thymus Mongolicus Ronn and the application thereof. The method comprises five technical links including obtaining of material, differentiation of adventive buds, rapid proliferation of cluster buds, rooting culture, and seeding exercising and transplant. Based on strict multiple comparative experiment researches, and through the unique innovation of culture conditions, components and proportions of transplant substrates, formulation of culture mediums for different culture stages, and the like, the survival rate can reach about 80%. The invention can be applied to establish a rapid propagation technique system of wild Thymus Mongolicus Ronn for artificial propagation, so that propagation of the wild Thymus Mongolicus Ronn can be accelerated, and the wild Thymus Mongolicus Ronn can be used as an excellent water-saving type landscaping ground cover plant.
Description
Technical field
The present invention relates to a kind ofly, relate in particular to a kind of wild Mongolian thyme tissue cultivation rapid breeding method and application wildly by the Plant Tissue Breeding fast breeding technique.
Background technology
The wild thyme aboundresources of China mainly is distributed in arids such as many tors ground about " three Norths " regional height above sea level 1100-3600m, the chiltern ground environment that faces south in China.Mongolia's thyme (Thymus mongolicus Ronn.) is Labiatae Thymus (Thymus) plant, perennial short and small fruticuli, and the field often is carpet-like and distributes, and branch, leaf, flower have strong fragrance.Drought resistance is very strong, can be used as good water-saving afforestation material and in the city trees and shrubs greenery patches, uses.
At present, the method that does not also have a kind of thymic quick breeding of wild Mongolia of practicality.
Summary of the invention
The purpose of this invention is to provide a kind of method and application of wild Mongolian thyme tissue culture quick breeding.
The objective of the invention is to realize through following technical scheme:
The method of wild Mongolian thyme tissue culture quick breeding of the present invention comprises step:
Condition of culture: 24 ± 2 ℃ of cultivation temperature, illuminance 3000 ± 100Lux, illumination 12 ± 1h.d
-1
The acquisition of sterilizable material: choose the wild Mongolian thyme plant of robust growth, no damage by disease and insect, the stem section of clip band axillalry bud is also carried out aseptic process, processes aseptic stem section;
The differentiation of indefinite bud: with the above-mentioned aseptic stem section of handling well, on superclean bench, cut into the long stem section that has 2～3 axillalry buds of 2～3cm, be inoculated on clump bud inducing culture, be cultured to the axillalry bud director and go out budlet, and differentiate a plurality of buds of growing thickly;
The bud fast breeding of growing thickly: when the bud length of growing thickly that differentiates is above to 1cm; The bud of will growing thickly is cut into every clump of 2 to 3 buds, is inoculated on the proliferated culture medium, forms the new bud of growing thickly; Again the new bud of growing thickly is inoculated into successive transfer culture on the identical proliferated culture medium, breeds the bud seedling of growing thickly;
Culture of rootage: the bud seedling of will growing thickly is inoculated into root induction on the bottled root media that film seals;
Refining seedling and transplanting: on root media, cultivated 26-27 days, and long to more than the 3cm, remove and sealed the spacious bottle refining of film seedling 3-4 days; Take out tissue cultivating seedling; With every clump of 5-7 strain is unit, after cleaning on the root medium and carrying out disinfection, is transplanted in the nutritive cube of the nutrition soil that fills sterilization and cultivates.
The application of the method for above-mentioned wild Mongolian thyme tissue culture quick breeding of the present invention, the wild Mongolian thyme that carries out artificial propagation through this method is as water-saving garden plants.
Technical scheme by the invention described above provides can be found out; The method and the application thereof of wild Mongolian thyme provided by the invention (Thymus mongolicus Ronn.) tissue culture quick breeding are owing to comprise the acquisition of sterilizable material, the differentiation of indefinite bud, the bud fast breeding of growing thickly, culture of rootage, refining seedling and transplant five sport technique segments.Multiple ratio through strictness is than experimental study, from the composition of prescription, condition of culture and the transplanting medium of the medium of different cultivation stages and proportioning etc. original innovation arranged, and survival rate can be about 80%.Use the present invention and set up wild Mongolian thyme fast breeding technique system, carry out artificial propagation, it is numerous to quicken the thymic expansion of wild Mongolia, can be used as good water-saving afforestation ground cover plant and uses.
Embodiment
To do to describe in detail further to the embodiment of the invention below.
The method of wild Mongolian thyme tissue culture quick breeding of the present invention, its preferable embodiment comprises step:
Condition of culture: 24 ± 2 ℃ of cultivation temperature, illuminance 3000 ± 100Lux, illumination 12 ± 1h.d
-1
The acquisition of sterilizable material: choose the wild Mongolian thyme plant of robust growth, no damage by disease and insect, the stem section of clip band axillalry bud is also carried out aseptic process, processes aseptic stem section;
The differentiation of indefinite bud: with the above-mentioned explant material of handling well; On superclean bench, cut into the long stem section that has 2～3 axillalry buds of 2～3cm, be inoculated on clump bud inducing culture, during the 4d of inoculation back; Axillalry bud obviously expands; When cultivating 15d, the axillalry bud director goes out budlet, and differentiates a plurality of buds of growing thickly;
The bud fast breeding of growing thickly: grow thickly bud length more than more than the 1cm when what differentiate; The bud of will growing thickly is cut into every clump of 2 to 3 buds, is inoculated on the proliferated culture medium, forms the new bud of growing thickly; Again the new bud of growing thickly is inoculated into successive transfer culture on the identical proliferated culture medium, breeds the bud seedling of growing thickly; Growth coefficient is 22.5.
Culture of rootage: the bud seedling of will growing thickly is inoculated into root induction on the bottled root media that film seals;
Refining seedling and transplanting: on root media, cultivated 26-27 days, and long to more than the 3cm, remove and sealed the spacious bottle refining of film seedling 3-4 days; Take out tissue cultivating seedling; With every clump of 5-7 strain is unit, after cleaning on the root medium and carrying out disinfection, is transplanted in the nutritive cube of the nutrition soil that fills sterilization and cultivates.
Said condition of culture specifically can for: 24 ± 2 ℃ of cultivation temperature, illuminance 3000Lux, light quality are fluorescent lamp, illumination 12h.d
-1
Specifically can comprise the stem section and the step of carrying out aseptic process in the procurement process of said sterilizable material: to the stem section flushing surface of band axillalry bud, use dish detergent solution soaking 10min then, behind the flowing water flushing 30min; On superclean bench; With 75% alcohol-pickled 30s sterilization, aseptic water washing 3 times soaks 8min with 0.1% mercuric chloride again; With aseptic water washing 5 times, cut away and the soup contact site.
In the step of the differentiation of said indefinite bud: the prescription of said clump bud inducing culture can be MS+ sucrose 30g/L+ agar 7g/L+6-BA 0.5mg/L+IAA0.1mg/L, pH5.8-6.0;
Cultivate in the said aseptic stem Duan Zaicong bud inducing culture after 21 days, each stem section can induce grows thickly bud quantity 5-10.
In the said bud fast breeding step of growing thickly: the prescription of said proliferated culture medium can be MS+ sucrose 30g/L+ agar 7g/L+6-BA 1mg/L+IAA0.5mg/L, pH5.8-6.0.
In the said bud fast breeding step of growing thickly: when growing thickly bud length more than more than the 1cm, the bud of will growing thickly is cut into every clump of 2 to 3 buds, is inoculated on the proliferated culture medium, can form the new bud of growing thickly.Inoculate successive transfer culture on the identical medium, can breed a large amount of buds of growing thickly, growth coefficient is 22.5.
In the said culture of rootage step: the prescription of said root media can be MS+ sucrose 30g/L+ agar 6g/L+IBA 0.7mg/L, pH5.8-6.0.
In said refining seedling and the transplant step:
To the sterilization method of the root of tissue cultivating seedling can for: in 0.5% liquor potassic permanganate, dip 5-10s;
The prescription of said nutrition soil can be the peat composed of rotten mosses: perlite: vermiculite=2: 1: 1, and said nutrition soil uses 0.5% disinfecting solution of potassium permanganate;
Cultivation condition can for: keep about 25 ± 2 ℃ of temperature, air humidity about 80 ± 2%, spray water 3-4 every day, became work in 13 to 15 days.
The application of the method for above-mentioned wild Mongolian thyme tissue culture quick breeding of the present invention, its preferable embodiment be, the wild Mongolian thyme that carries out artificial propagation through this method is as water-saving garden plants.
Specific embodiment:
Condition of culture: 24 ± 2 ℃ of cultivation temperature, illuminance 3000Lux, light quality are fluorescent lamp, illumination 12h.d
-1
The acquisition of sterilizable material: choose the plant of robust growth, no damage by disease and insect, the stem section of clip band axillalry bud, flushing surface; Dish detergent solution soaking 10min is behind the flowing water flushing 30min, on superclean bench; With 75% alcohol-pickled 30s sterilization, aseptic water washing 3 times soaks 8min with 0.1% mercuric chloride again; With aseptic water washing 5 times, cut away and the soup contact site, subsequent use.
The differentiation of indefinite bud: aseptic stem section is inoculated on clump bud inducing culture; The prescription of medium is: MS+ sucrose 30g/L+ agar 7g/L+6-BA 0.5mg/L+IAA0.1mg/L; PH5.8-6.0 cultivated after 14 days, and the axillalry bud director goes out budlet; Differentiated the more bud of growing thickly in 21 days, each stem section can induce grows thickly bud quantity 5-10.
The bud fast breeding of growing thickly: when growing thickly bud length more than more than the 1cm; The bud of will growing thickly is cut into every clump of 2 to 3 buds, is inoculated on the proliferated culture medium, and the prescription of medium is: MS+ sucrose 30g/L+ agar 7g/L+6-BA 1mg/L+IAA0.5mg/L; PH5.8-6.0 can form the new bud of growing thickly.Inoculate successive transfer culture on the identical medium, can breed a large amount of buds of growing thickly, growth coefficient is 22.5.
Culture of rootage: with the seedling of high 1-2cm, be inoculated into root induction on the root media, the prescription of medium is: MS+ sucrose 30g/L+ agar 6g/L+IBA 0.7mg/L, and pH5.8-6.0 began to take root after 4 days, and rooting rate reaches 100%.
Refining seedling and transplanting: on root media, cultivated 26-27 days, long to more than the 3cm, remove and sealed the spacious bottle refining of film seedling 3-4 days; Taking out tissue cultivating seedling, is unit (whenever from the 5-7 strain) with the clump, cleans medium on the root; In 0.5% liquor potassic permanganate, dip 5-10s, be transplanted to the nutrition soil (peat composed of rotten mosses: perlite: in nutritive cube vermiculite=2: 1: 1), keep about 25 ℃ of temperature that fills with 0.5% disinfecting solution of potassium permanganate; About air humidity 80%; Spray water 3-4 every day, becomes to live survival rate about 80% about 14 days.
The method and the application thereof of wild Mongolian thyme of the present invention (Thymus mongolicus Ronn.) tissue culture quick breeding comprise the acquisition of sterilizable material, the differentiation of indefinite bud, the bud fast breeding of growing thickly, culture of rootage, refining seedling and transplant five sport technique segments.Multiple ratio through strictness is than experimental study, from the composition of prescription, condition of culture and the transplanting medium of the medium of different cultivation stages and proportioning etc. original innovation arranged, and survival rate can be about 80%.Use the present invention and set up wild Mongolian thyme fast breeding technique system, carry out artificial propagation, it is numerous to quicken the thymic expansion of wild Mongolia, can be used as good water-saving afforestation ground cover plant and uses.
The above; Be merely the preferable embodiment of the present invention, but protection scope of the present invention is not limited thereto, any technical staff who is familiar with the present technique field is in the technical scope that the present invention discloses; The variation that can expect easily or replacement all should be encompassed within protection scope of the present invention.Therefore, protection scope of the present invention should be as the criterion with the protection domain of claims.
Claims (8)
1. the method for a wild Mongolian thyme tissue culture quick breeding is characterized in that, comprises step:
Condition of culture: 24 ± 2 ℃ of cultivation temperature, illuminance 3000 ± 100Lux, illumination 12 ± 1h.d
-1
The acquisition of sterilizable material: choose the wild Mongolian thyme plant of robust growth, no damage by disease and insect, the stem section of clip band axillalry bud is also carried out aseptic process, processes aseptic stem section;
The differentiation of indefinite bud: with the above-mentioned aseptic stem section of handling well, on superclean bench, cut into the long stem section that has 2～3 axillalry buds of 2～3cm, be inoculated on clump bud inducing culture, be cultured to the axillalry bud director and go out budlet, and differentiate a plurality of buds of growing thickly;
The bud fast breeding of growing thickly: when the bud length of growing thickly that differentiates is above to 1cm; The bud of will growing thickly is cut into every clump of 2 to 3 buds, is inoculated on the proliferated culture medium, forms the new bud of growing thickly; Again the new bud of growing thickly is inoculated into successive transfer culture on the identical proliferated culture medium, breeds the bud seedling of growing thickly;
Culture of rootage: the bud seedling of will growing thickly is inoculated into root induction on the bottled root media that film seals;
Refining seedling and transplanting: on root media, cultivated 26-27 days, and long to more than the 3cm, remove and sealed the spacious bottle refining of film seedling 3-4 days; Take out tissue cultivating seedling; Being unit whenever, after cleaning on the root medium and carrying out disinfection, being transplanted in the nutritive cube of the nutrition soil that fills sterilization and cultivating from the 5-7 strain.
2. the method for wild Mongolian thyme tissue culture quick breeding according to claim 1 is characterized in that, said condition of culture is specially: 24 ± 2 ℃ of cultivation temperature, illuminance 3000Lux, light quality are fluorescent lamp, illumination 12h.d
-1
3. the method for wild Mongolian thyme tissue culture quick breeding according to claim 1 is characterized in that, in the procurement process of said sterilizable material the stem section and the step of carrying out aseptic process is specifically comprised: to the stem section flushing surface of band axillalry bud; Use dish detergent solution soaking 10min then, behind the flowing water flushing 30min, on superclean bench; With 75% alcohol-pickled 30s sterilization, aseptic water washing 3 times soaks 8min with 0.1% mercuric chloride again; With aseptic water washing 5 times, cut away and the soup contact site.
4. the method for wild Mongolian thyme tissue culture quick breeding according to claim 1; It is characterized in that; In the step of the differentiation of said indefinite bud: the prescription of said clump bud inducing culture is: MS+ sucrose 30g/L+ agar 7g/L+6-BA 0.5mg/L+IAA0.1mg/L, pH5.8-6.0;
Cultivate in the said aseptic stem Duan Zaicong bud inducing culture after 21 days, each stem section induces grows thickly bud quantity 5-10.
5. the method for wild Mongolian thyme tissue culture quick breeding according to claim 1; It is characterized in that; In the said bud fast breeding step of growing thickly: the prescription of said proliferated culture medium is: MS+ sucrose 30g/L+ agar 7g/L+6-BA1mg/L+IAA0.5mg/L, pH5.8-6.0.
6. the method for wild Mongolian thyme tissue culture quick breeding according to claim 1 is characterized in that, in the said culture of rootage step: the prescription of said root media is: MS+ sucrose 30g/L+ agar 6g/L+IBA0.7mg/L, pH5.8-6.0.
7. the method for wild Mongolian thyme tissue culture quick breeding according to claim 1 is characterized in that, in said refining seedling and the transplant step:
Sterilization method to the root of tissue cultivating seedling is: in 0.5% liquor potassic permanganate, dip 5-10s;
The prescription of said nutrition soil is: the peat composed of rotten mosses: perlite: vermiculite=2: 1: 1, and said nutrition soil is used 0.5% disinfecting solution of potassium permanganate;
Cultivation condition is: keep about 25 ± 2 ℃ of temperature, and air humidity about 80 ± 2%, spray water 3-4 every day, became to live in 13 to 15 days.
8. the application of the method for each described wild Mongolian thyme tissue culture quick breeding of claim 1 to 7 is characterized in that, the wild Mongolian thyme that carries out artificial propagation through this method is as water-saving garden plants.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104381134A (en) * | 2014-12-08 | 2015-03-04 | 邱林 | Rapid mongollian thyme herb proliferation method |
| CN104782494A (en) * | 2015-05-02 | 2015-07-22 | 冯文杰 | Thymus mongolicus Ronn. tissue culture rapid propagation method |
| CN105532451A (en) * | 2015-12-03 | 2016-05-04 | 深圳职业技术学院 | Method for inducing plant regeneration with thymus serpyllum aseptic seedlings |
| CN109197596A (en) * | 2018-11-21 | 2019-01-15 | 黑龙江省科学院火山与矿泉研究所 | Tissue culture and rapid propagation method of wild thymus serpyllum |
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2011
- 2011-10-27 CN CN2011103324672A patent/CN102388807A/en active Pending
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| 《内蒙古林业科技》 20090930 赵颖等 百里香的组织培养及快繁技术 第63页左栏第1行至第64页右栏第1行 1－6 第35卷, 第3期 * |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104381134A (en) * | 2014-12-08 | 2015-03-04 | 邱林 | Rapid mongollian thyme herb proliferation method |
| CN104381134B (en) * | 2014-12-08 | 2016-11-30 | 湖北云天花海旅游股份有限公司 | A kind of method of Herba Thymi fast breeding |
| CN104782494A (en) * | 2015-05-02 | 2015-07-22 | 冯文杰 | Thymus mongolicus Ronn. tissue culture rapid propagation method |
| CN105532451A (en) * | 2015-12-03 | 2016-05-04 | 深圳职业技术学院 | Method for inducing plant regeneration with thymus serpyllum aseptic seedlings |
| CN109197596A (en) * | 2018-11-21 | 2019-01-15 | 黑龙江省科学院火山与矿泉研究所 | Tissue culture and rapid propagation method of wild thymus serpyllum |
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Application publication date: 20120328 |