CN106165648B - A kind of cercis tissue culture culture medium and cultural method - Google Patents
A kind of cercis tissue culture culture medium and cultural method Download PDFInfo
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- CN106165648B CN106165648B CN201610719546.1A CN201610719546A CN106165648B CN 106165648 B CN106165648 B CN 106165648B CN 201610719546 A CN201610719546 A CN 201610719546A CN 106165648 B CN106165648 B CN 106165648B
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- cercis
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- culture medium
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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
-
- 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
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
-
- 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
Abstract
The present invention relates to technical field of tissue culture, specifically discloses a kind of cercis tissue culture culture medium and cercis quickly tissue culture propagation method.The nutrient media components includes：The D of wpm, 0.4~0.6mg/L 2.4,0.2~0.4mg/L IBA, 0.4~0.6mg/L KT；Preferably：The D+0.3mg/L IBA+0.5mg/L KT+30g/L sucrose+5g/L agar of wpm+0.5mg/L 2.4.During tissue culture propagation, cercis stem with bud is taken, the breeding of cercis, strong sprout can be carried out simultaneously with taking root using culture medium of the present invention, you can quick to obtain cercis tissue-cultured seedling, the cercis tissue-cultured seedling of gained is not required to special hardening, robust growth, and survival rate is high.The tissue culture propagation of cercis is carried out by the method for the invention, can not only keep the genetic stability of fine tree species, and its breeding coefficient can be effectively improved, breeding cycle is short, and hardening process easily survives, and reduces domestication cost, it is easy to operate, laid the foundation further to carry out breed improvement.
Description
Technical field
The present invention relates to technical field of tissue culture, specifically, is related to a kind of cercis tissue culture culture medium and cultural method.
Background technology
Cercis (Cercis chinensis Bunge), pulse family Redbud, deciduous tree or shrub.Originate in China.Happiness
Illumination, there is certain cold resistance.Manurial-philous fertile, well-drained soil, intolerant to flooding.Sprout tillers is strong, resistance to trimming.Follicarpium wood flower all may be used
It is used as medicine, its seed is poisonous.It is family and the beautiful, symbol of bone and flesh deep love.Its morphological feature has at a relatively high Landscape Application value,
It is more rare and excellent decorative flower, there is very high economic value.
At present, cercis breeding is concentrated mainly on seed propagation and cutting propagation, but both breeding practices are by season
Limitation, and seed propagation germination percentage is low, the seed dormancy time is longer, and cutting propagation efficiency is low.
The content of the invention
In order to solve problems of the prior art, it is an object of the invention to provide a kind of cercis tissue culture culture medium and training
The method of supporting.
In order to realize the object of the invention, technical scheme is as follows：
In a first aspect, the invention provides a kind of cercis tissue culture culture medium, the nutrient media components includes：Wpm, 0.4~
0.6mg/L 2.4-D, 0.2~0.4mg/L IBA (indolebutyric acid), 0.4~0.6mg/L KT (kinetin).
The culture medium can directly induce cercis stem section to take root, while by inducing shoot proliferation, only pass through the culture
Base can a step realize the quick breeding of cercis stem section.
Preferably, the nutrient media components includes：wpm+0.5mg/L 2.4-D+0.3mg/L IBA+0.5mg/L KT.
Further, the nutrient media components also includes sucrose and agar.
More preferably, the culture medium is：wpm+0.5mg/L 2.4-D+0.3mg/L IBA+0.5mg/L KT+30g/L
Sucrose+5g/L agar.
The present invention has found that the culture medium of the formula can more preferably realize the quick of cercis stem section faster by experimental study
Breeding.It is embodied in：Subculture cycle is 30d, rootage duration 7d；The adventitious buds proliferation cycle is 30d, and Multiple Buds simple bud is high by 1
~3cm, growth coefficient are 2~6.
Further, screened through assay optimization, pH=5.4~6.0 of the culture medium are advisable, preferably pH=6.0.
Second aspect, cercis quickly tissue culture propagation method is carried out using prior culture media the invention provides one kind.
Specially：Cercis stem with bud is taken, propagation culture of rootage is carried out using prior culture media of the present invention, you can obtain purple
Chaste tree tissue-cultured seedling.
The cercis tissue-cultured seedling of gained is not required to special hardening, robust growth.By the cercis tissue-cultured seedling of stalwartness of taking root, remnants are washed off
Culture medium after, directly transplant into warmhouse booth seedbed, pay attention to shading cooling moisture-retaining, survival rate is up to 100%.It is preferred that transplant
Matrix is humus mixing perlite, and mixed proportion is preferably 2:1.
In the method for the invention, propagation and the rooting process of cercis stem with bud can the same steppings in the culture medium
OK, need to prepare culture medium respectively and carry out propagation and culture of rootage respectively that there is significant difference with prior art.
Further, the condition of culture for carrying out breeding culture of rootage in prior culture media is intensity of illumination 2800LX, illumination
Time 16h, cultivate in the culturing room that 24~28 DEG C of temperature, as intensity of illumination is less than 2800LX, takes root slowly or do not take root, be higher than
There is yellow leaf in 2800LX, blade, and temperature is slow-growing less than 24 DEG C or higher than 28 DEG C, or even the plant phenomena of mortality occurs.
Further, it is foregoing to take cercis stem with bud, preferably using the sterile tissue-cultured seedling of cercis of stalwartness as female parent, take stem segment with bud
Section, carries out the propagation culture of rootage.
The sterile tissue-cultured seedling of healthy and strong cercis can be obtained by conventional culture methods, it also may be preferable for be obtained by the following method
：
The preparation method of the sterile tissue-cultured seedling of cercis comprises the following steps：
S1, by the cercis seed after sterilizing, be inoculated in germination medium, in 2700~2900lx of intensity of illumination (preferably
2800lx), 15~17h of light application time (preferably 16h), cultivate 2-3 weeks under conditions of 22~25 DEG C of temperature (preferably 24 DEG C), obtain
Seedling；
S2, seedling is cut be transferred in Initial culture base, in 2700~2900lx of illumination (preferably 2800lx), illumination
15~17h of time (preferably 16h), cultivate 2 weeks under conditions of 22~25 DEG C of temperature (preferably 24 DEG C), acquisition is taken root sterile group of cercis
Train seedling.
Wherein, preferably, the germination medium is：MS+ sucrose 20g/L+ agar 5g/L, pH5.5~6.0；
The Initial culture base is：1/2MS+0.5mg/L NAA+ sucrose 30g/L+ agar 5g/L, pH5.5~6.0.
Further, the cercis seed after the sterilizing can obtain by the following method：
8~October cercis seed maturity, fine day is afternoon, first gather cercis no disease and pests harm healthy and strong branch on full kind
Son, dry in the shade naturally；Cercis pericarp is removed with hand rubbing afterwards, it is standby to choose full seed.Seed is invaded into 1~3h of bubble with sterile warm water,
Blot excessive moisture with filter paper, be put on superclean bench, with 75% alcohol sterilize 30s, twice of aseptic water washing, 0.1% liter
Mercury sterilizes 10min, then with sterile water wash six times.It is standby on superclean bench after blotting excessive moisture with filter paper.
The beneficial effects of the present invention are：
It can make cercis stem with bud the invention provides one kind while be bred, strong sprout, the training for the special formulation taken root
Support base.Merely with the culture medium stem section can be induced to take root, while by sprouting shoot proliferation, so as to realize the quickly tissue culture of cercis
Breeding.
The tissue culture propagation of cercis is carried out by the method for the invention, can not only keep the genetic stability of fine tree species,
And its breeding coefficient can be effectively improved, the breeding cycle is short, and hardening process easily survives, and reduces domestication cost, easy to operate, can
It is relatively easy to and obtains cercis aseptic seedling, be the effective way of cercis tissue-culturing rapid propagation, base is established further to carry out breed improvement
Plinth, strong technical support is provided for industrial seedling rearing.
Brief description of the drawings
Fig. 1 is that the cercis primary seed in experimental example 1 of the present invention is sprouted；
Fig. 2 is the cercis one-step method subculture (breed-take root) in experimental example 1 of the present invention；
Fig. 3 is the cercis rooted seedling in experimental example 1 of the present invention；
Fig. 4 is the cercis root system in experimental example 1 of the present invention；
Fig. 5 is 2 years after the cercis hardening in experimental example 1 of the present invention survives.
Embodiment
The preferred embodiment of the present invention is described in detail below in conjunction with embodiment.It is it will be appreciated that following real
Providing merely to play the purpose of explanation for example is applied, is not used to limit the scope of the present invention.The skill of this area
Art personnel can carry out various modifications and replacement in the case of without departing substantially from spirit of the invention and spirit to the present invention.
Experimental method used in following embodiments is conventional method unless otherwise specified.
Material used, reagent etc., unless otherwise specified, are commercially obtained in following embodiments.
The culture medium of embodiment 1
The specific formula of culture medium described in the present embodiment is：wpm+0.5mg/L 2.4-D+0.3mg/L IBA+0.5mg/L
KT+30g/L sucrose+5g/L agar.
The culture medium of embodiment 2
The present embodiment and the difference of embodiment 1 are：Specific formula is：wpm+0.4mg/L2.4-D+0.3mg/L IBA+
0.5mg/L KT+30g/L sucrose+5g/L agar.
The culture medium of embodiment 3
The present embodiment and the difference of embodiment 1 are：Specific formula is：wpm+0.5mg/L2.4-D+0.4mg/L IBA+
0.5mg/L KT+30g/L sucrose+5g/L agar.
The culture medium of embodiment 4
The present embodiment and the difference of embodiment 1 are：Specific formula is：wpm+0.5mg/L2.4-D+0.3mg/L IBA+
0.4mg/L KT+30g/L sucrose+5g/L agar.
The culture medium of embodiment 5
The present embodiment and the difference of embodiment 1 are：Specific formula is：wpm+0.5mg/L2.4-D+0.3mg/L IBA+
0.5mg/L KT+10g/L sucrose+5g/L agar.
The culture medium of comparative example 1
The difference of this comparative example and embodiment 1 is：Wpm is replaced with into MS.
The culture medium of comparative example 2
The difference of this comparative example and embodiment 1 is：KT is replaced with into NAA.
Experimental example 1
This experimental example is used to illustrate cercis quickly tissue culture method of the present invention, specific as follows：
1st, by the cercis seed after sterilizing, it is inoculated in germination medium, in illumination 2800lx, light application time 16h, temperature
Cultivated 2-3 weeks under conditions of 24 DEG C, obtain seedling；
2nd, seedling is cut and be transferred in Initial culture base, in illumination 2800lx, light application time 16h, 24 DEG C of temperature
Under the conditions of cultivate 2 weeks, acquisition is taken root the sterile tissue-cultured seedling of cercis (as shown in Figure 1).
3rd, the sterile tissue-cultured seedling of cercis is cut into 3-6 section stem with bud, using the culture medium described in embodiment 1, is light
According to 2800LX, light application time 16h carries out propagation culture of rootage under conditions of 24 DEG C of temperature, and after culture 2 weeks, stem with bud grows up to
Intact plant (as shown in Figure 2), and plant takes root flourishing (as shown in Figure 3), root system is sturdy, dense (as shown in Figure 4).
More than, the breeding of cercis quickly tissue culture is completed.
Still health, growing state are good (as shown in Figure 5) after the cercis plant of tissue culture propagation survives 2 years.
Experimental example 2
This experimental example is for the culture medium described in comparing embodiment 1~5 and comparative example 1~2 on tissue culture propagation cercis
Difference, it is specific as follows：
Cercis stem with bud is taken, embodiment 1~5 and the culture medium described in comparative example 1~2 is respectively adopted, is illumination
2800LX, light application time 16h, propagation culture of rootage is carried out under conditions of 24 DEG C of temperature.
It is 22-30 DEG C that cercis tissue-cultured seedling, which is recorded, in cultivation temperature, and intensity of illumination is 2500-2800Lx, and light application time is
14—18h/d.Wherein primary culture medium uses WPM minimal mediums, supplementation with growth hormones, sucrose, agar.Trained substantially in WPM
Under conditions of supporting base, sucrose, agar all same, it is grouped according to growth hormone concentration and number of days, observes and record cultivating seedling
Situation, specific culture situation the results are shown in Table 1 each item data：
The cercis of table 1 grows statistical conditions
As shown in Table 1, culture medium described in embodiment 1 take root situation and plant growing way is substantially better than embodiment 2~5.
Rootage duration is most short in embodiment 1, and radical is most, and can germinate new leaves piece three for seven days, and plant plant height can reach within 30 days
4cm, it is sufficient to prove and growing way is preferable, several and average root long of averagely taking root is higher, is the most suitable growth of root media
Hormone Conditions.Culture medium described in comparative example 1 only has effect to subculture plant height, and yellow leaf is more, and growing way is poor, can not realize life
Root.Culture medium described in comparative example 2, which can not be realized, takes root.
As can be seen here, the formula of culture medium of the present invention has obvious advantage in terms of quickly tissue culture breeds cercis.
Although above the present invention is described in detail with a general description of the specific embodiments,
On the basis of the present invention, it can be made some modifications or improvements, this will be apparent to those skilled in the art.Cause
This, these modifications or improvements, belong to the scope of protection of present invention without departing from theon the basis of the spirit of the present invention.
Claims (3)
1. a kind of cercis quickly tissue culture propagation method, it is characterised in that using the sterile tissue-cultured seedling of cercis of stalwartness as female parent, take its band
Leaf stem section, propagation culture of rootage is carried out using cercis tissue culture culture medium, produces cercis tissue-cultured seedling；
The condition of culture is to be cultivated in intensity of illumination 2800LX, light application time 16h, the culturing room that 24 DEG C of temperature；
The cercis tissue culture culture medium is：Wpm+0.5mg/L 2.4-D+0.3mg/L IBA+0.5mg/L KT+30g/L sucrose+
5g/L agar, the pH value of the culture medium is 5.4~6.0.
2. according to the method for claim 1, it is characterised in that the preparation method of the sterile tissue-cultured seedling of cercis includes as follows
Step：
S1, by the cercis seed after sterilizing, be inoculated in germination medium, in 2700~2900lx of intensity of illumination, light application time
15~17h, cultivate 2-3 weeks under conditions of 22~25 DEG C of temperature, obtain seedling；
S2, seedling is cut be transferred in Initial culture base, in 2700~2900lx of illumination, 15~17h of light application time, temperature
Cultivated 2 weeks under conditions of 22~25 DEG C, obtain the sterile tissue-cultured seedling of cercis of taking root.
3. according to the method for claim 2, it is characterised in that the germination medium is：MS+ sucrose 20g/L+ agar
5g/L, pH5.5~6.0；The Initial culture base is：1/2MS+0.5mg/L NAA+ sucrose 30g/L+ agar 5g/L, pH5.5~
6.0。
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| CN106665353B (en) * | 2016-12-08 | 2019-07-19 | 上海杉一植物科技有限公司 | A kind of subculture method of huge cercis tissue-cultured seedling |
| CN114027196B (en) * | 2021-12-07 | 2022-07-12 | 钦州市林业科学研究所 | Tissue culture and rapid propagation culture medium combination and culture method of cercis fascicularis |
| CN115997684B (en) * | 2022-12-30 | 2023-10-13 | 广州市林业和园林科学研究院 | Culture medium and culture method for tissue culture and seedling raising of cercis chinensis |
Citations (3)
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| CN103355170A (en) * | 2013-07-24 | 2013-10-23 | 江苏绿苑园林建设有限公司 | Tissue culture method of cerciscanadensis forestpansy |
| CN104782490A (en) * | 2015-05-02 | 2015-07-22 | 冯文杰 | Establishment method of Cercis Canadensis 'Forest Pansy' tissue culture rapid propagation technical system |
| CN105638461A (en) * | 2015-12-30 | 2016-06-08 | 四川禾木本业农林科技有限公司 | Method for preventing browning of callus of toxicodendron succedaneum |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103355170A (en) * | 2013-07-24 | 2013-10-23 | 江苏绿苑园林建设有限公司 | Tissue culture method of cerciscanadensis forestpansy |
| CN104782490A (en) * | 2015-05-02 | 2015-07-22 | 冯文杰 | Establishment method of Cercis Canadensis 'Forest Pansy' tissue culture rapid propagation technical system |
| CN105638461A (en) * | 2015-12-30 | 2016-06-08 | 四川禾木本业农林科技有限公司 | Method for preventing browning of callus of toxicodendron succedaneum |
Non-Patent Citations (1)
| Title |
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| 紫叶加拿大紫荆组织培养研究;孟月娥等;《河南农业科学》;20101231(第12期);摘要、第1.1、1.6节 * |
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Address after: 621000 Huaishu Village Group 6, Dongyu Town, Nanjiang County, Bazhong City, Sichuan Province Patentee after: Sichuan Qicai Forestry Co., Ltd. Address before: 621000 Huaishu Village Group 6, Dongyu Town, Nanjiang County, Bazhong City, Sichuan Province Patentee before: SICHUAN QICAI FORESTRY INDUSTRY DEVELOPMENT CO., LTD. |
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Address after: 635600 Tissue Culture Workshop Building of Rare and Colorful Leaf Plant Varieties in Guangwu Mountain, Sanshe, Changtan Village, Zhengzheng Town, Nanjiang County, Bazhong City, Sichuan Province Patentee after: Sichuan Qicai Forestry Co., Ltd. Address before: 621000 Huaishu Village Group 6, Dongyu Town, Nanjiang County, Bazhong City, Sichuan Province Patentee before: Sichuan Qicai Forestry Co., Ltd. |