CN102265787A - Tissue culture method of president clematis - Google Patents
Tissue culture method of president clematis Download PDFInfo
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- CN102265787A CN102265787A CN2010101931233A CN201010193123A CN102265787A CN 102265787 A CN102265787 A CN 102265787A CN 2010101931233 A CN2010101931233 A CN 2010101931233A CN 201010193123 A CN201010193123 A CN 201010193123A CN 102265787 A CN102265787 A CN 102265787A
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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 ' clematis.
Background technology
Clematis is a Ranunculaceae Clematis plant, originates in China, is widely distributed in the Northern Hemisphere, kind surplus the original seed 200, and a large amount of garden-varieties appear in the artificial hybridization through for many years, and pattern is clearly simple and elegant, spend big look aquatic foods, and the florescence is long, can reach November from May successively because of the kind difference.The scene of blooming magnificence is welcome by common people, is present domestic very fast-selling garden liana material.
Clematis ' president ' is a garden-variety, and woody climber is about 1-2 rice.Brown or the aubergine of stem, single leaf is often to life; Flower Dan Sheng, purple.The florescence 6-9 month.Be the excellent new varieties that obtain by artificially breeding, because its plant division speed is slow, the cottage propagation rate is low, and kind is easily degenerated, and can't satisfy a large amount of demand in market.Some enterprises directly sell from external import bareroot seedling, the cost height, 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 ' 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 clematis is characterized in that, this method may further comprise the steps:
(1) obtains aseptic raw material: wash 2h with running water after winning clematis ' president ' tender shoots, be the alcohol immersion 25-35s of 75wt% successively with concentration on superclean bench, 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 of bud and propagation: tender shoots is inoculated in the bud inducing culture after 5 weeks, bastem portion begins to expand and the yellow green projection occurs, continue to cultivate the visible significantly callus in 3 week backs, cultivated 1 month again, the callus that downcuts the band bud is inoculated on the adventitious bud proliferation medium;
(3) strong seedling culture of indefinite bud: the bud of growing thickly that on the adventitious bud proliferation medium, induces, 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) culture of rootage: get the indefinite bud plant of 2-3cm, with root induction in its root media of transferring, the seedling base section dissolves the root original hase of many whites after 10 days, and the root original hase grows to 4-6cm after 30 days;
(5) plant hardening and transplanting: the root original hase that step (4) obtains continued culture of rootage 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 and is given rich water quality management after 40 days in the greenhouse, can obtain 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 medium 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 medium is 5.6-6.0, and cultivation 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 medium.
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 regularity of seedling propagation speed and seedling, and keeps original maternal character better by tissue culture technology, 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 ' clematis, this method may further comprise the steps:
(1) obtains aseptic raw material: wash 2h with running water after winning clematis ' president ' tender shoots, be the alcohol immersion 25s of 75wt% successively with concentration on superclean bench, 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 of bud and propagation: tender shoots is inoculated in the bud inducing culture after 5 weeks, bastem portion begins to expand and the yellow green projection occurs, continue to cultivate the visible significantly callus in 3 week backs, cultivated again 1 month, the callus that downcuts the band bud is inoculated on the adventitious bud proliferation medium, the composition of adventitious bud proliferation medium is MS+6-BA2.0mg/L+NAA0.2mg/L, though the base portion callus is many, but do not influence the propagation of indefinite bud, the indefinite bud growth there is no bad phenomenon such as vitrifying rapidly, and it is good to grow in this cultivation;
(3) strong seedling culture of indefinite bud: the bud of growing thickly that on the adventitious bud proliferation medium, induces, 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) culture of rootage: the indefinite bud plant 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 root original hase of many whites after 10 days, the root original hase grows to 4cm after 30 days, and fibrous root is numerous;
(5) plant hardening and transplanting: the root original hase that step (4) obtains continued culture of rootage 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 and is given rich water quality management after 40 days in the greenhouse, can obtain clematis ' president ' plant.
Above medium component also comprises sucrose 30g/L, agar 6g/L, and the pH value of this medium is 5.6, and the control cultivation temperature is 24 ℃ during use, and illumination condition adopts 70 μ mol/ms.
Embodiment 2
The method of a kind of tissue culture ' president ' clematis, this method may further comprise the steps:
(1) obtains aseptic raw material: wash 2h with running water after winning clematis ' president ' tender shoots, be the alcohol immersion 35s of 75wt% successively with concentration on superclean bench, 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 of bud and propagation: tender shoots is inoculated in the bud inducing culture after 5 weeks, bastem portion begins to expand and the yellow green projection occurs, continue to cultivate the visible significantly callus in 3 week backs, cultivated again 1 month, the callus that downcuts the band bud is inoculated on the adventitious bud proliferation medium, the composition of adventitious bud proliferation medium is MS+6-BA3.0mg/L+NAA0.3mg/L, though the base portion callus is many, but do not influence the propagation of indefinite bud, the indefinite bud growth there is no bad phenomenon such as vitrifying rapidly, and it is good to grow in this cultivation;
(3) strong seedling culture of indefinite bud: the bud of growing thickly that on the adventitious bud proliferation medium, induces, 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) culture of rootage: the indefinite bud plant 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 root original hase of many whites after 10 days, the root original hase grows to 6cm after 30 days, and fibrous root is numerous;
(5) plant hardening and transplanting: the root original hase that step (4) obtains continued culture of rootage 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 and is given rich water quality management after 40 days in the greenhouse, can obtain clematis ' president ' plant.
Above medium component also comprises sucrose 30g/L, agar 6g/L, and the pH value of this medium is 5.6, and the control cultivation temperature is 26 ℃ during use, and illumination condition adopts 90 μ mol/ms.
Embodiment 3
The method of a kind of tissue culture ' president ' clematis, this method may further comprise the steps:
(1) obtains aseptic raw material: wash 2h with running water after winning clematis ' president ' tender shoots, be the alcohol immersion 30s of 75wt% successively with concentration on superclean bench, 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 of bud and propagation: tender shoots is inoculated in the bud inducing culture after 5 weeks, bastem portion begins to expand and the yellow green projection occurs, continue to cultivate the visible significantly callus in 3 week backs, cultivated again 1 month, the callus that downcuts the band bud is inoculated on the adventitious bud proliferation medium, the composition of adventitious bud proliferation medium is MS+6-BA1.0mg/L+NAA0.1mg/L, though the base portion callus is many, but do not influence the propagation of indefinite bud, the indefinite bud growth there is no bad phenomenon such as vitrifying rapidly, and it is good to grow in this cultivation;
(3) strong seedling culture of indefinite bud: the bud of growing thickly that on the adventitious bud proliferation medium, induces, 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) culture of rootage: the indefinite bud plant 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 root original hase of many whites after 10 days, the root original hase grows to 6cm after 30 days, and fibrous root is numerous;
(5) plant hardening and transplanting: the root original hase that step (4) obtains continued culture of rootage 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 and is given rich water quality management after 40 days in the greenhouse, can obtain clematis ' president ' plant.
Above medium component also comprises sucrose 30g/L, agar 6g/L, and the pH value of this medium is 5.68, and the control cultivation temperature is 25 ℃ during use, and illumination condition adopts 80 μ mol/ms.
Claims (6)
1. the method for a tissue culture president clematis is characterized in that, this method may further comprise the steps:
(1) obtains aseptic raw material: wash 2h with running water after winning clematis ' president ' tender shoots, be the alcohol immersion 25-35s of 75wt% successively with concentration on superclean bench, 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 of bud and propagation: tender shoots is inoculated in the bud inducing culture after 5 weeks, bastem portion begins to expand and the yellow green projection occurs, continue to cultivate the visible significantly callus in 3 week backs, cultivated 1 month again, the callus that downcuts the band bud is inoculated on the adventitious bud proliferation medium;
(3) strong seedling culture of indefinite bud: the bud of growing thickly that on the adventitious bud proliferation medium, induces, 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) culture of rootage: get the indefinite bud plant of 2-3cm, with root induction in its root media of transferring, the seedling base section dissolves the root original hase of many whites after 10 days, and the root original hase grows to 4-6cm after 30 days;
(5) plant hardening and transplanting: the root original hase that step (4) obtains continued culture of rootage 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 and is given rich water quality management after 40 days in the greenhouse, can obtain clematis ' president ' plant.
2. the method for a kind of tissue culture president clematis according to claim 1, it is characterized in that 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.
3. the method for a kind of tissue culture president clematis according to claim 1, it is characterized in that the composition of described adventitious bud proliferation medium 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.
4. the method for a kind of tissue culture president clematis according to claim 1, it is characterized in that 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.
5. the method for a kind of tissue culture president clematis according to claim 1, it is characterized in that 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.
6. according to claim 2 or 3 or the methods of 4 or 5 described a kind of tissue culture president clematis, it is characterized in that, described medium component also comprises sucrose 30g/L, agar 6g/L, the pH value of this medium is 5.6-6.0, cultivation temperature is 24-26 ℃, and illumination condition adopts 70-90 μ mol/ms.
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Cited By (11)
| 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 |
| CN103430854A (en) * | 2013-09-22 | 2013-12-11 | 南京林业大学 | Tissue culturing method of clematis guernsey cream |
| CN103461131A (en) * | 2013-09-22 | 2013-12-25 | 南京林业大学 | Tissue culture method for clematis Betty Risdon |
| CN103461130A (en) * | 2013-09-22 | 2013-12-25 | 江苏农林职业技术学院 | Tissue culture method for changeable protea of clematis cultivated variety |
| 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 |
| 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 |
| 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 |
| 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 |
| CN108184662A (en) * | 2016-12-08 | 2018-06-22 | 上海植物园 | The high-efficiency in-vitro quick-breeding method and its culture medium of a kind of clematis |
| CN113207687A (en) * | 2021-05-18 | 2021-08-06 | 西南林业大学 | Tissue culture and rapid propagation method for clematis |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| 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 |
| CN103461131B (en) * | 2013-09-22 | 2015-06-03 | 南京林业大学 | Tissue culture method for clematis Betty Risdon |
| CN103461130A (en) * | 2013-09-22 | 2013-12-25 | 江苏农林职业技术学院 | Tissue culture method for changeable protea of clematis cultivated variety |
| CN103461131A (en) * | 2013-09-22 | 2013-12-25 | 南京林业大学 | Tissue culture method for clematis Betty Risdon |
| 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 |
| 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 |
| CN108184662A (en) * | 2016-12-08 | 2018-06-22 | 上海植物园 | The high-efficiency in-vitro quick-breeding method and its culture medium of a kind of clematis |
| 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 |
| CN113207687A (en) * | 2021-05-18 | 2021-08-06 | 西南林业大学 | Tissue culture and rapid propagation method for clematis |
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