CN102265787B - Tissue culture method of president clematis - Google Patents
Tissue culture method of president clematis Download PDFInfo
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- CN102265787B CN102265787B CN 201010193123 CN201010193123A CN102265787B CN 102265787 B CN102265787 B CN 102265787B CN 201010193123 CN201010193123 CN 201010193123 CN 201010193123 A CN201010193123 A CN 201010193123A CN 102265787 B CN102265787 B CN 102265787B
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Abstract
The invention relates to a tissue culture method of president clematis. Tender shoots of 'president' clematis are selected as a raw material; and a clematis 'president' plant is acquired through steps of an aseptic processing, a differentiation and propagation of tender shoots, a strong seedling culture of adventitious buds, a rooting culture of a root, a plant hardening seedling and transplanting, etc. Compared with a prior art, the invention employs tissue culture to increase a breeding speed of nursery stock and regularity of the seedlings, to better maintain original female parent properties, and to realize factory seedling growing, so as to guarantee market supply demand.
Description
Technical field
The present invention relates to a kind of plant tissue culture method, especially relate to the method for a kind of tissue culture ' president ' Root of Cream Clematis.
Background technology
Root of Cream Clematis is the Ranunculaceae Actions of Clematis Species, originates in China, is widely distributed in the Northern Hemisphere, and original seed more than 200 is planted, and a large amount of garden-varieties appear in the artificial hybridization through for many years, pattern clearly simple and elegant, spend large look bright, the florescence is long, can reach successively from May November because of the kind difference.The scene of blooming magnificence is welcome by common people, is present domestic very fast-selling garden liana material.
Root of Cream Clematis ' president ' is garden-variety, and woody climber is about 1-2 rice.Brown or the red-purple of stem, single leaf is often to life; Flower Dan Sheng, purple.The florescence 6-9 month.Be the excellent new variety that obtain by artificially breeding, because its plant division speed is slow, the cottage propagation rate is low, and kind is easily degenerated, can't a large amount of demand of satisfying the market.Some enterprises directly sell from external import bare-root seeding, and cost is high, and profit is little, and therefore, how improving output and reducing cultivation is a difficult problem that needs to be resolved hurrily now with cost.
Summary of the invention
Purpose of the present invention is exactly to provide a kind of original maternal character of holding in order to overcome the defective that above-mentioned prior art exists, the method for tissue culture ' president ' Root of Cream Clematis of the batch production that realizes growing seedlings.
Purpose of the present invention can be achieved through the following technical solutions:
The method of a kind of tissue culture president Root of Cream Clematis is characterized in that, the method may further comprise the steps:
(1) obtains aseptic starting material: wash 2h with tap water after winning Root of Cream Clematis ' president ' tender shoots, be the alcohol immersion 25-35s of 75wt% successively with concentration on Bechtop, the mercuric chloride of 1w ‰ soaks 10-15min, blot surperficial moisture content with behind aseptic water washing 5-6 time, get the tender shoots base portion, be cut into 1cm and be inoculated on the bud inducing culture after long;
(2) Differentiation and proliferation of bud: tender shoots is inoculated in the bud inducing culture after 5 weeks, bastem section begins to expand and the yellow-green colour projection occurs, continue to cultivate rear visible significantly callus of 3 weeks, cultivated 1 month again, the callus that downcuts with bud is inoculated on the adventitious bud proliferation substratum;
(3) strong seedling culture of indefinite bud: the Multiple Buds that induces at the adventitious bud proliferation substratum, every clump has 2-3 strain elongation, is inoculated on the strong seedling culture base after being divided into Xiao Cong or simple bud, and indefinite bud can be grown 2-3cm after 20 days;
(4) root culture: get the adventitious bud plants of 2-3cm, with root induction in its root media of transferring, the seedling base section dissolves the former base of root of many whites after 10 days, and the former base of root grows to 4-6cm after 30 days;
(5) plant hardening and transplanting: the former base of the root that step (4) obtains continued root culture 20-30 days, the aseptic seedling of selecting the well developed root system robust growth was in indoor uncork hardening 5 days, take out afterwash root agar, domestication is transplanted outdoor after 40 days and is given rich water quality management in the greenhouse, can obtain Root of Cream Clematis ' president ' plant.
The composition of described bud inducing culture is MS+6-BA1.0mg/L+NAA0.1mg/L, MS+6-BA3mg/L+NAA0.3mg/L or MS+6-BA5.0mg/L+NAA0.5mg/L.
The composition of described adventitious bud proliferation substratum is MS+6-BA1.0mg/L+NAA0.1mg/L, MS+6-BA2.0mg/L+NAA0.2mg/L or MS+6-BA3.0mg/L+NAA0.3mg/L.
The composition of described strong seedling culture base is MS+6-BA0.5mg/L+NAA0.1mg/L, MS+6-BA1.0mg/L+NAA0.1mg/L or MS+6-BA2.0mg/L+NAA0.2mg/L.
The composition of described root media is MS+NAA0.1mg/L+IBA1.0mg/L, MS+NAA0.3mg/L+IBA3.0mg/L or MS+NAA0.5mg/L+IBA5.0mg/L.
Described medium component also comprises sucrose 30g/L, agar 6g/L, and the pH value of this substratum is 5.6-6.0, and culture temperature is 24-26 ℃, and illumination condition adopts 70-90 μ mol/ms.
In addition, the preferred MS+6-BA3.0mg/L+NAA0.3mg/L of the composition of described bud inducing culture.
The preferred MS+6-BA1.0mg/L+NAA0.1mg/L of the composition of described adventitious bud proliferation substratum.
The preferred MS+6-BA0.5mg/L+NAA0.1mg/L of the composition of described strong seedling culture base.
The preferred MS+NAA0.3mg/L+IBA3.0mg/L of the composition of described root media.
Compared with prior art, the present invention improves the reguarity of seedling propagation speed and seedling, and keeps better original maternal character by tissue culture technique, realizes growing seedlings batch production, thereby can guarantee the market supply demand.
Embodiment
The present invention is described in detail below in conjunction with specific embodiment.
Embodiment 1
The method of a kind of tissue culture ' president ' Root of Cream Clematis, the method may further comprise the steps:
(1) obtains aseptic starting material: wash 2h with tap water after winning Root of Cream Clematis ' president ' tender shoots, be the alcohol immersion 25s of 75wt% successively with concentration on Bechtop, the mercuric chloride of 1w ‰ soaks 10min, blot surperficial moisture content with behind the aseptic water washing 5 times, get the tender shoots base portion, be inoculated on the bud inducing culture after being cut into 1cm length, the composition of bud inducing culture is MS+6-BA1.0mg/L+NAA0.1mg/L;
(2) Differentiation and proliferation of bud: tender shoots is inoculated in the bud inducing culture after 5 weeks, bastem section begins to expand and the yellow-green colour projection occurs, continue to cultivate rear visible significantly callus of 3 weeks, cultivated again 1 month, cutting-out is inoculated on the adventitious bud proliferation substratum with the callus of bud, the composition of adventitious bud proliferation substratum is MS+6-BA2.0mg/L+NAA0.2mg/L, although the base portion callus is many, but do not affect the propagation of indefinite bud, the indefinite bud growth there is no rapidly the bad phenomenon such as vitrifying, grows in this cultivation good;
(3) strong seedling culture of indefinite bud: the Multiple Buds that induces at the adventitious bud proliferation substratum, every clump has 2 strains elongation, all the other are in the dwarfing state, be inoculated on the strong seedling culture base after being divided into Xiao Cong or simple bud, indefinite bud can be grown 2cm after 20 days, the strong seedling culture base consist of MS+6-BA1.0mg/L+NAA0.1mg/L;
(4) root culture: the adventitious bud plants of getting 2cm, with root induction in its root media of transferring, the composition of this root media is MS+NAA0.1mg/L+IBA1.0mg/L, and the seedling base section dissolves the former base of root of many whites after 10 days, the former base of root grows to 4cm after 30 days, and fibrous root is numerous;
(5) plant hardening and transplanting: the former base of the root that step (4) obtains continued root culture 20 days, the aseptic seedling of selecting the well developed root system robust growth was in indoor uncork hardening 5 days, take out afterwash root agar, domestication is transplanted outdoor after 40 days and is given rich water quality management in the greenhouse, can obtain Root of Cream Clematis ' president ' plant.
Above medium component also comprises sucrose 30g/L, agar 6g/L, and the pH value of this substratum is 5.6, and the control culture temperature is 24 ℃ during use, and illumination condition adopts 70 μ mol/ms.
Embodiment 2
The method of a kind of tissue culture ' president ' Root of Cream Clematis, the method may further comprise the steps:
(1) obtains aseptic starting material: wash 2h with tap water after winning Root of Cream Clematis ' president ' tender shoots, be the alcohol immersion 35s of 75wt% successively with concentration on Bechtop, the mercuric chloride of 1w ‰ soaks 15min, blot surperficial moisture content with behind the aseptic water washing 6 times, get the tender shoots base portion, be inoculated on the bud inducing culture after being cut into 1cm length, the composition of bud inducing culture is MS+6-BA5.0mg/L+NAA0.5mg/L;
(2) Differentiation and proliferation of bud: tender shoots is inoculated in the bud inducing culture after 5 weeks, bastem section begins to expand and the yellow-green colour projection occurs, continue to cultivate rear visible significantly callus of 3 weeks, cultivated again 1 month, cutting-out is inoculated on the adventitious bud proliferation substratum with the callus of bud, the composition of adventitious bud proliferation substratum is MS+6-BA3.0mg/L+NAA0.3mg/L, although the base portion callus is many, but do not affect the propagation of indefinite bud, the indefinite bud growth there is no rapidly the bad phenomenon such as vitrifying, grows in this cultivation good;
(3) strong seedling culture of indefinite bud: the Multiple Buds that induces at the adventitious bud proliferation substratum, every clump has 3 strains elongation, all the other are in the dwarfing state, be inoculated on the strong seedling culture base after being divided into Xiao Cong or simple bud, indefinite bud can be grown 3cm after 20 days, the strong seedling culture base consist of MS+6-BA2.0mg/L+NAA0.2mg/L;
(4) root culture: the adventitious bud plants of getting 3cm, with root induction in its root media of transferring, the composition of this root media is MS+NAA0.5mg/L+IBA5.0mg/L, and the seedling base section dissolves the former base of root of many whites after 10 days, the former base of root grows to 6cm after 30 days, and fibrous root is numerous;
(5) plant hardening and transplanting: the former base of the root that step (4) obtains continued root culture 30 days, the aseptic seedling of selecting the well developed root system robust growth was in indoor uncork hardening 5 days, take out afterwash root agar, domestication is transplanted outdoor after 40 days and is given rich water quality management in the greenhouse, can obtain Root of Cream Clematis ' president ' plant.
Above medium component also comprises sucrose 30g/L, agar 6g/L, and the pH value of this substratum is 5.6, and the control culture temperature is 26 ℃ during use, and illumination condition adopts 90 μ mol/ms.
Embodiment 3
The method of a kind of tissue culture ' president ' Root of Cream Clematis, the method may further comprise the steps:
(1) obtains aseptic starting material: wash 2h with tap water after winning Root of Cream Clematis ' president ' tender shoots, be the alcohol immersion 30s of 75wt% successively with concentration on Bechtop, the mercuric chloride of 1w ‰ soaks 15min, blot surperficial moisture content with behind the aseptic water washing 6 times, get the tender shoots base portion, be inoculated on the bud inducing culture after being cut into 1cm length, the composition of bud inducing culture is MS+6-BA3.0mg/L+NAA0.3mg/L;
(2) Differentiation and proliferation of bud: tender shoots is inoculated in the bud inducing culture after 5 weeks, bastem section begins to expand and the yellow-green colour projection occurs, continue to cultivate rear visible significantly callus of 3 weeks, cultivated again 1 month, cutting-out is inoculated on the adventitious bud proliferation substratum with the callus of bud, the composition of adventitious bud proliferation substratum is MS+6-BA1.0mg/L+NAA0.1mg/L, although the base portion callus is many, but do not affect the propagation of indefinite bud, the indefinite bud growth there is no rapidly the bad phenomenon such as vitrifying, grows in this cultivation good;
(3) strong seedling culture of indefinite bud: the Multiple Buds that induces at the adventitious bud proliferation substratum, every clump has 3 strains elongation, all the other are in the dwarfing state, be inoculated on the strong seedling culture base after being divided into Xiao Cong or simple bud, indefinite bud can be grown 3cm after 20 days, the strong seedling culture base consist of MS+6-BA0.5mg/L+NAA0.1mg/L;
(4) root culture: the adventitious bud plants of getting 3cm, with root induction in its root media of transferring, the composition of this root media is MS+NAA0.3mg/L+IBA3.0mg/L, and the seedling base section dissolves the former base of root of many whites after 10 days, the former base of root grows to 6cm after 30 days, and fibrous root is numerous;
(5) plant hardening and transplanting: the former base of the root that step (4) obtains continued root culture 25 days, the aseptic seedling of selecting the well developed root system robust growth was in indoor uncork hardening 5 days, take out afterwash root agar, domestication is transplanted outdoor after 40 days and is given rich water quality management in the greenhouse, can obtain Root of Cream Clematis ' president ' plant.
Above medium component also comprises sucrose 30g/L, agar 6g/L, and the pH value of this substratum is 5.68, and the control culture temperature is 25 ℃ during use, and illumination condition adopts 80 μ mol/ms.
Claims (1)
1. one kind is used for the substratum that presidential Root of Cream Clematis tissue is cultivated, it is characterized in that comprising bud inducing culture, adventitious bud proliferation substratum, strong seedling culture base and root media, wherein, the composition of described bud inducing culture is MS+6-BA5.0mg/L+NAA0.5mg/L+ sucrose 30g/L+ agar 6g/L; The composition of described adventitious bud proliferation substratum is MS+6-BA3.0mg/L+NAA0.3mg/L+ sucrose 30g/L+ agar 6g/L; The composition of described strong seedling culture base is MS+6-BA2.0mg/L+NAA0.2mg/L+ sucrose 30g/L+ agar 6g/L; The composition of described root media is MS+NAA0.5mg/L+IBA5.0mg/L+ sucrose 30g/L+ agar 6g/L.
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Families Citing this family (11)
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| 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 |
| CN103461130B (en) * | 2013-09-22 | 2015-05-20 | 江苏农林职业技术学院 | Tissue culture method for changeable protea of clematis cultivated variety |
| CN103461131B (en) * | 2013-09-22 | 2015-06-03 | 南京林业大学 | Tissue culture method for clematis Betty Risdon |
| CN103430854A (en) * | 2013-09-22 | 2013-12-11 | 南京林业大学 | Tissue culturing method of clematis guernsey cream |
| CN104082137A (en) * | 2014-06-26 | 2014-10-08 | 江苏农林职业技术学院 | Tissue culture method of clematis cultivar Violet Elizabeth |
| CN104082152A (en) * | 2014-08-04 | 2014-10-08 | 云南农业大学 | Tissue culture and rapid propagation method for clematis ranunculoides |
| CN104082151A (en) * | 2014-08-04 | 2014-10-08 | 云南农业大学 | Cultivation method for polyploid clematis ranunculoides |
| CN107864855A (en) * | 2016-09-27 | 2018-04-03 | 上海上房园艺有限公司 | A kind of method of the fast breeding iron chopsticks based on tissue cultures |
| CN108184662B (en) * | 2016-12-08 | 2021-09-03 | 上海植物园 | Efficient in-vitro rapid propagation method and culture medium of clematis |
| CN107864865A (en) * | 2017-12-26 | 2018-04-03 | 丽江市古城区秋成种养殖有限公司 | A kind of efficient hardening technology and domesticating cultivation method of haw ginseng tissue-cultured seedling |
| CN113207687B (en) * | 2021-05-18 | 2022-06-10 | 西南林业大学 | Tissue culture and rapid propagation method for clematis |
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| CN1868263A (en) * | 2006-06-29 | 2006-11-29 | 南京林业大学 | Tissue culture method for Clematis Multi Blue |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1868263A (en) * | 2006-06-29 | 2006-11-29 | 南京林业大学 | Tissue culture method for Clematis Multi Blue |
Non-Patent Citations (2)
| Title |
|---|
| 张启香.观赏型铁线莲的引种及生物学研究.《中国博士学位论文全文数据库（农业科技辑）》.2008,(第2期), * |
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Effective date of registration: 20140108 Address after: 201114 Pujiang village, Pujiang Town, Shanghai, Minhang District Patentee after: Gardening Co., Ltd. Shanghai never ending Patentee after: State Grid Shanghai Municipal Electric Power Company Patentee after: Shanghai Urban Power Supply Design Co., Ltd. Address before: 201114 Pujiang village, Pujiang Town, Shanghai, Minhang District Patentee before: Gardening Co., Ltd. Shanghai never ending |