CN102144543A - Tissue culture and rapid propagation technology for Clematis 'Arabella' - Google Patents
Tissue culture and rapid propagation technology for Clematis 'Arabella' Download PDFInfo
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- CN102144543A CN102144543A CN 201110000713 CN201110000713A CN102144543A CN 102144543 A CN102144543 A CN 102144543A CN 201110000713 CN201110000713 CN 201110000713 CN 201110000713 A CN201110000713 A CN 201110000713A CN 102144543 A CN102144543 A CN 102144543A
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Abstract
The invention discloses a tissue culture and rapid propagation technology for Clematis 'Arabella', which belongs to the technical field of tissue culture and rapid propagation of the Clematis 'Arabella'. In the technology, a regeneration system of the Clematis 'Arabella' is established by taking a stem segment, which is provided with buds, of a new branch in the current year as an explant. The research discovers that: the optimal culture medium for axillary bud induction comprises a murashige and skoog (MS) culture medium, 2,4-dichlorophenoxyacetic acid at the concentration of 1.0mg/L, indolebutyric acid at the concentration of 1.0mg/L and sucrose at the concentration of 30g/L, and the differentiation rate reaches 100 percent; the optimal culture medium for adventitious bud proliferation comprises an MS culture medium, 6-benzyladenine at the concentration of 0.5mg/L, indolebutyric acid at the concentration of 0.1mg/L and sucrose at the concentration of 30g/L, and the proliferation coefficient is 3.98; and an appropriate rooting culture medium comprises a 1/2MS culture medium and naphthylacetic acid at the concentration of 0.04mg/L, and the rooting rate reaches 89.3 percent. Through 5 to 7 days of bottle-opening seedling hardening, pearlite and peat in a ratio of 1:3 are taken as matrices, and after 20 days, the investigation discovers that the transplant survival rate of seedlings reaches 90 percent. The technology can be applied to the efficient and rapid propagation of the Clematis 'Arabella', and can realize the industrial breeding of the Clematis 'Arabella'.
Description
One, technical field
The present invention relates to clematis ' Arabela ' tissue culture rapid propagating technology.
Two, background technology
Clematis ' Arabela ' (Clematis ' Arabella ') is under the jurisdiction of Ranunculaceae (Ranunculaceae) Clematis (Clematis L.), be the Hybrid of clematis Clematis ' Integrifolia ' and Clematis ' Lanujinosa ', the former Britain of originating from.This kind plant flowers type, inflorescence are various, and pattern is abundant, and the florescence is long, and horticultural use is extensive, and ornamental value is quite high.' Arabela ' not only is applied to the field, gardens as the vertical greening material, and be widely used in potted plant, cut-flower etc.
Clematis ' Arabela ' ornamental value is higher, and application prospect is wide, but because of its seminal propagation difficulty, promotion rate is slow.China does not also work out its fast breeding technique at present, and this kind still needs to have limited ' Arabela ' promotion and application in China from external a large amount of imports, also consumes a large amount of foreign exchanges of China simultaneously.So research ' Arabela ' fast breeding technique is extremely necessary.
Clematis is wide in variety, but also few for the Study on tissue culture of clematis so far, the seedling that the prosperous nurse of damp benevolence utilizes the seed of Herba Clematidis tanguticae to grow has carried out the generation of indefinite bud and adventive root; The best explant that Zhang Qixiang has found out ' Multi-Blue ' is a stem apex; And Zhang Tao induces the callus of polyphyll clematis, but does not have evoking adventive bud.Present technique is utilized clematis ' Arabela ' innovation bar stem with bud then, carries out tissue culture, induces the generation of axillalry bud, promotes the generation of indefinite bud, induces it to take root again.By artificial preparation Different Nutrition composition and hormone regulating and controlling, ' Arabela ' parent pattern is gorgeous keeping, on the basis of characteristics such as the flower type is big, form thousands of plantlets, obtain a large amount of test-tube plantlets in short time, the seedling that obtains is deposited and can be placed in the manually operated culturing room, can realize that batch production clone throughout the year grows seedlings.Therefore utilize tissue culture technique breeding clematis ' Arabela ', not only provide an approach fast for promoting clematis ' Arabela ', also produce for its batch production and offer help, application prospect is long-range.
Three, summary of the invention
The purpose of this invention is to provide a kind of can the realization fast and the tissue culture technique of a large amount of breeding clematis ' Arabela '.
Clematis provided by the invention ' Arabela ' tissue culture rapid propagating technology comprises following step:
1, axillalry bud is induced:
Innovation bar stem with bud is inoculated into inducing culture then, and this medium contains 1/4MS, 1/2MS, the conventional minimal medium of MS, 2,4 dichlorophenoxyacetic acid 1.0 mg/litre, indolebutyric acid 0.1 mg/litre, sucrose 30 grams per liters.Cultivate after 20 days, treat that culture is induced the original hase that sprouts after, enter following the 2nd the step.
2, adventitious bud proliferation:
The culture of the 1st step gained is transferred on the proliferated culture medium, and proliferated culture medium contains the conventional minimal medium of MS, 6-benzyl purine 0.5-1.5 mg/litre, indolebutyric acid 0.05-0.15 mg/litre, sucrose 20-40 grams per liter.Cultivated about 20 days, and treated that seedling was long when routine is taken root the program desired height, enter the following the 3rd and go on foot.
3, take root:
The neat root of unrooted seedling of the 2nd step gained is taken off, go in the conventional root media, the minimal medium of taking root contains 1/4MS, 1/2MS, the conventional minimal medium of MS, methyl 0.05 mg/litre were cultivated about 20 days, treated that the seedling root grows short root, can carry out the elongation of root, the root elongation medium contains the conventional minimal medium of 1/2MS, methyl 0-1.0 mg/litre or indolebutyric acid 0.03-1.0 mg/litre, sucrose 30 grams per liters.About 35 days, when seedling grows 4-5 root main root, enter following the 4th step.
4, transplanting and hardening:
Open sealing film earlier, hardening is taken out seedling about 1 week in culturing room, cleans the agar of seedling root, is transplanted to perlite and peat with in the matrix of mixing at 1: 3, puts into the further hardening 10-15 of green house days.Need to spray 1/10MS macro-element nutrients liquid therebetween and be beneficial to alleviate rising dehydration, and additional plant nutrition.
The present invention utilizes the stem section of band axillalry bud to do the generation that explant carries out direct organ, has improved fast numerous speed of tissue culture greatly, adds micro-hormone with minimal medium simultaneously and carries out inducing of axillalry bud, can the rapid induction axillalry bud, and impel its elongation.After axillalry bud is induced, should not be on former medium long-term cultivation, otherwise therefore the easy elongation growth of the bud that induces will transfer on MS+6-benzyl purine 0.5 mg/litre+indolebutyric acid 0.1 mg/litre+sucrose 30 grams per liter medium, is beneficial to the differentiation and the elongation of indefinite bud.Take root the stage, the present invention has adopted and has induced the method that promotes elongation more earlier, at first, be fit to the take root seedling of height of routine being elongated to, transfer in the 1/2MS+ methyl 0.03 mg/litre medium, the generation of inducing adventitious root is transferred in the promotion root elongation 1/2MS+ methyl 0.04 mg/litre medium again, and the method has improved the speed of root induction and elongation greatly.
Therefore adopt the medium of this invention that following advantage is arranged: axillalry bud is induced and is started soon, bud coefficient of differentiation height, and the seedling elongation is fast, growth is consistent and healthy and strong, rooting rate height, root length and strong, growth cycle is short, does not have lopsided seedling substantially.
Four, description of drawings
Figure 12,4-D and IBA induce the axillalry bud of generation
Propagation bud after Figure 26-BA and the IBA HORMONE TREATMENT
Fig. 3 NAA induces the root of generation
Five, embodiment
The present invention will be further described below in conjunction with embodiment.
Embodiment
1, draw materials: give birth to spray then with clematis ' Arabela ' and cultivate as explant, require the innovation robust growth, blade has maternal merit, and has full and axillalry bud that do not sprout.
2, materials disinfection method: remove unnecessary blade, only keep the stem section, clean the fine hair of removing on the explant gently, flowing water flushing 1 hour with alcohol absorbent cotton.Under aseptic condition, with 75% alcohol-pickled 30 seconds, 0.1% mercuric chloride was handled 3-5 minute, aseptic water washing 5～6 times.
3, inoculation: in superclean bench, after aseptic filter paper is drawn residual moisture, stem section two is cut a bit of, to reduce the murder by poisoning of mercuric chloride.Be cut into the long stem section of 1～1.5cm then, be inoculated into the explant induction medium then.Inducing culture consists of: MS+2,4-dichlorphenoxyacetic acid 1.0 mg/litre+indolebutyric acid 0.1 mg/litre+sucrose 30 grams per liters.
4, indefinite bud clump propagation: stem segment with axillary bud is inoculated into inducing culture just can sees after last 20 day that axillary bud sprouting grows tall (Fig. 1), then it is transferred on the proliferated culture medium.Proliferated culture medium consists of: MS+6-benzyl purine 0.5 mg/litre+indolebutyric acid 0.1 mg/litre+sucrose 30 grams per liters.
5, take root: long when routine is taken root desired height when the bud of growing thickly (Fig. 2) that differentiates, cut with scissors is single, be inoculated in the root media.Consisting of of root media: 1/2MS+ methyl 0.03 mg/litre.
For making the root that induces can better elongation growth, the seedling of will taking root be transferred in the root elongation medium, the consisting of of root elongation medium: 1/2MS+ methyl 0.04 mg/litre (Fig. 3).
Cultivation temperature and illumination condition: temperature is 25 ± 1 ℃; Illumination is 2000 luxs, 16 hours/day.
6, transplanting and hardening: the seedling of taking root growth is after about 35 days, and root system is relatively more flourishing, in the time of generally can growing 4-5 bar main root, can transplant.
Open sealing film earlier, hardening is taken out seedling about 1 week in culturing room, cleans the agar of seedling root, is transplanted to perlite and peat with in the matrix of mixing at 1: 3, puts into the further hardening 10-15 of green house days.Need to spray 1/10MS macro-element nutrients liquid therebetween and be beneficial to alleviate rising dehydration, and additional plant nutrition, in case the jaundice of uppermost leaf sheet.
Claims (5)
1. a clematis ' Arabela ' tissue culture rapid propagating technology is characterized in that following four parts form:
(1) axillalry bud is induced: incite somebody to action the aseptic stem section of innovation band bud then, be inoculated into inducing culture, this medium contains 1/4MS, 1/2MS, the conventional minimal medium, 2 of MS, 4-dichlorphenoxyacetic acid 1.0 mg/litre, indolebutyric acid 0.1 mg/litre, sucrose 30 grams per liters, cultivate after 20 days, treat that culture induces the original hase that sprouts, can begin the cultivation of next step;
(2) adventitious bud proliferation: the culture of the 1st step gained is transferred on the proliferated culture medium, proliferated culture medium contains the conventional minimal medium of MS, 6-benzyl purine 0.5-1.5 mg/litre, indolebutyric acid 0.05-0.15 mg/litre, sucrose 20-40 grams per liter, cultivated about 20 days, and entered following the 3rd step;
(3) take root: the neat root of unrooted seedling of the 2nd step gained is taken off, go in the conventional root media, the minimal medium of taking root contains 1/4MS, 1/2MS, the conventional minimal medium of MS, methyl 0.03 mg/litre, cultivated about 20 days, treat that the seedling root grows short root, can carry out the elongation of root, the root elongation medium is: the conventional minimal medium of 1/2MS, methyl 0-1.0 mg/litre or indolebutyric acid 0.03-1.0 mg/litre, sucrose 30 grams per liters, about 35 days, can transplant and grow up to normal clematis plant;
(4) transplanting and hardening: when treating that the 3rd step, seedling grew 4-5 root main root, open sealing film earlier, hardening is about 1 week in culturing room, take out seedling, clean the agar of seedling root, be transplanted to perlite and peat, put into the further hardening 10-15 of green house days with in the matrix of mixing at 1: 3, need to spray 1/10MS macro-element nutrients liquid therebetween and be beneficial to alleviate rising dehydration, and additional plant nutrition.
2. induce by the axillalry bud in the described clematis of claim 1 ' Arabela ' the tissue culture quick breeding process, it is characterized in that described inducing culture is: 1/4MS, 1/2MS, the conventional minimal medium of MS, 2,4 dichlorophenoxyacetic acid 1.0 mg/litre, indolebutyric acid 0.1 mg/litre, sucrose 30 grams per liters.
3. by the adventitious bud proliferation in the described clematis of claim 1 ' Arabela ' the tissue culture quick breeding process, it is characterized in that described proliferated culture medium is: the conventional minimal medium of MS, 6-benzyl purine 0.5-1.5 mg/litre, indolebutyric acid 0.05-0.15 mg/litre, sucrose 20-40 grams per liter.
4. by taking root in the described clematis of claim 1 ' Arabela ' the tissue culture quick breeding process, it is characterized in that described root media is: the conventional minimal medium of 1/2MS, methyl 0-1.0 mg/litre or indolebutyric acid 0.03-1.0 mg/litre, sucrose 30 grams per liters.
5. by transplanting and hardening in the described clematis of claim 1 ' Arabela ' the tissue culture quick breeding process, it is characterized in that described grafting matrix is: perlite: peat=1: 3.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103202229A (en) * | 2013-04-11 | 2013-07-17 | 福建农林大学 | Tissue culturing and rapid propagating method for chloranthy florida var. plena |
| CN109601388A (en) * | 2019-01-29 | 2019-04-12 | 中国林业科学研究院林业研究所 | A kind of quick breeding method for tissue culture hybridizing clematis |
| CN109618927A (en) * | 2018-12-21 | 2019-04-16 | 滁州学院 | A kind of method for tissue culture of sweetautumn clematis |
| CN110235781A (en) * | 2018-03-09 | 2019-09-17 | 东北林业大学 | Clematis Blekitny Aniol tissue cultures and Regeneration System |
| CN112005886A (en) * | 2020-10-11 | 2020-12-01 | 江苏东郁植物科技有限公司 | Breeding method for clematis seedling tissue culture |
| CN114391475A (en) * | 2022-01-26 | 2022-04-26 | 江苏农林职业技术学院 | Tissue culture method of clematis krispipe angel |
| CN114946659A (en) * | 2022-06-09 | 2022-08-30 | 江苏省中国科学院植物研究所 | In-vitro rapid propagation method of clematis brachypodium |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101869070A (en) * | 2009-04-24 | 2010-10-27 | 上海上房园林植物研究所 | Tissue culture method of pink champagne clematis |
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2011
- 2011-01-05 CN CN 201110000713 patent/CN102144543A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101869070A (en) * | 2009-04-24 | 2010-10-27 | 上海上房园林植物研究所 | Tissue culture method of pink champagne clematis |
Non-Patent Citations (1)
| Title |
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| 《中国博士学位论文全文数据库 农业科技辑》 20080215 张启香 观赏型铁线莲的引种及生物学研究 第34页最后一段；第35页第1-3行；第36页第3行；第50页第1.2.2.6.4节第2行 1-5 , 第2期 * |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103202229A (en) * | 2013-04-11 | 2013-07-17 | 福建农林大学 | Tissue culturing and rapid propagating method for chloranthy florida var. plena |
| CN103202229B (en) * | 2013-04-11 | 2014-04-09 | 福建农林大学 | Tissue culturing and rapid propagating method for chloranthy florida var. plena |
| CN110235781A (en) * | 2018-03-09 | 2019-09-17 | 东北林业大学 | Clematis Blekitny Aniol tissue cultures and Regeneration System |
| CN109618927A (en) * | 2018-12-21 | 2019-04-16 | 滁州学院 | A kind of method for tissue culture of sweetautumn clematis |
| CN109601388A (en) * | 2019-01-29 | 2019-04-12 | 中国林业科学研究院林业研究所 | A kind of quick breeding method for tissue culture hybridizing clematis |
| CN109601388B (en) * | 2019-01-29 | 2022-03-22 | 中国林业科学研究院林业研究所 | Tissue culture rapid propagation method of hybrid clematis |
| CN112005886A (en) * | 2020-10-11 | 2020-12-01 | 江苏东郁植物科技有限公司 | Breeding method for clematis seedling tissue culture |
| CN114391475A (en) * | 2022-01-26 | 2022-04-26 | 江苏农林职业技术学院 | Tissue culture method of clematis krispipe angel |
| CN114946659A (en) * | 2022-06-09 | 2022-08-30 | 江苏省中国科学院植物研究所 | In-vitro rapid propagation method of clematis brachypodium |
| CN114946659B (en) * | 2022-06-09 | 2023-09-08 | 江苏省中国科学院植物研究所 | In-vitro rapid propagation method of clematis with short columns |
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Application publication date: 20110810 |