CN101869070A - Tissue culture method of pink champagne clematis - Google Patents
Tissue culture method of pink champagne clematis Download PDFInfo
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- CN101869070A CN101869070A CN200910049978A CN200910049978A CN101869070A CN 101869070 A CN101869070 A CN 101869070A CN 200910049978 A CN200910049978 A CN 200910049978A CN 200910049978 A CN200910049978 A CN 200910049978A CN 101869070 A CN101869070 A CN 101869070A
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Abstract
The invention relates to a tissue culture method of pink champagne clematis. The method comprises the following steps of: obtaining a sterile material; differentiating and proliferating buds; seedling and culturing adventitious buds; rooting and culturing; acclimatizing and transplanting; and the like. Compared with the prior art, by using the method, the reproduction rate and the seed uniformity of the pink champagne clematis are greatly improved, original female characters are well kept, and the modern industrialized mass production of seedlings can be realized.
Description
Technical field
The present invention relates to the method for tissue culture of a plant species, especially relate to a kind of method for tissue culture of pink champagne 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, a large amount of garden-varieties appear in the artificial hybridization through for many years, pattern refreshing clean simple and elegant, spend big look bright and numerous phase is of a specified duration, almost can reach November successively from May.The scene of blooming magnificence is welcome by common people, is good garden material.Pink champagne clematis is a garden-variety, and woody climber is about 1-2 rice, and stem is brown or aubergine, single leaf, and often to life, flower Dan Sheng, pink, florescence 6-9 month.But obtain excellent new varieties by seed selection, plant division speed is slow, the cottage propagation rate is low, kind is easily degenerated, and therefore can't satisfy people's demand.
Summary of the invention
Purpose of the present invention is exactly to provide a kind of regularity that improves reproduction speed and seedling for the defective that overcomes above-mentioned prior art existence, and keeps the method for tissue culture of the pink champagne clematis of original maternal character.
Purpose of the present invention can be achieved through the following technical solutions:
The method for tissue culture of pink champagne clematis is characterized in that, this method may further comprise the steps:
(1) acquisition of sterilizable material
Get the bud on the young shoot of sprouting spring, with behind the running water flushing 1-3h on ultra-clean work team workbench, utilize the alcohol immersion 10-50s of mass concentration successively for 60-75%, volumetric concentration is that the mercury of 0.5-2 ‰ soaks 10-30min, use aseptic water washing 4-6 time again, after utilizing aseptic filter paper to blot the moisture on bud surface, bud is cut into the sections of the long band axillalry bud of 0.5-2cm, is inoculated on the axillalry bud inducing culture;
(2) differentiation of bud and propagation
The sections of band axillalry bud is inoculated on the axillalry bud inducing culture, 1-3 began to expand in the axillalry bud position after week, the green-yellow projection appears, 3-5 is visible bud meristematic tissue after week, cultivates 1-2 month again, and axillalry bud is long to 3-5cm, with axillalry bud cutting-out carrying out enrichment culture, the bud of differentiation changes in the adventitious bud proliferation medium, though the base portion of the bud of differentiation has more callus, does not influence the propagation of indefinite bud;
(3) indefinite bud strong seedling culture
On the adventitious bud proliferation medium, every clump has the 2-4 strain to extend in the bud of growing thickly that induces, and remaining bud of growing thickly is in the dwarfing state, after the bud of will growing thickly is divided into Xiao Cong, go on the strong seedling culture base and grow, wherein the indefinite bud elongation can be grown to 3-5cm after 15-30 days rapidly;
(4) culture of rootage
Get the plantlet of 2-3cm, root induction in the root media is gone in switching, and the seedling base section dissolves the root original hase of white after 5-15 days, begins to take root after 18-25 days, can grow to 1-2cm after 30-35 days, and every strain has 3-4 root, and rooting rate is 80-90%;
(5) hardening and transplanting
Culture of rootage 30-35 days, when root system grows to 1-2cm, select the aseptic seedling of well developed root system, robust growth, in indoor uncork hardening 1-3 days, then seedling is taken out, clean the agar of root, plant in the seedbed in greenhouse and tamed 20-40 days, can transplant basin, give rich water quality management, final transplanting survival rate is 90-95%.
Described axillalry bud inducing culture comprises MS+6-BA1.0-3.0mg/L+NAA0.05-0.2mg/L.
The preferred MS+6-BA2.0mg/L+NAA0.2mg/L of described axillalry bud inducing culture.
Described adventitious bud proliferation medium comprises MS+6-BA1.0-3.0mg/L+NAA0.05-0.2mg/L.
The preferred MS+6-BA1.0mg/L+NAA0.2mg/L of described adventitious bud proliferation medium.
Described strong seedling culture base comprises MS+6-BA0.5-2.0mg/L+NAA0.1-0.2mg/L.
The preferred MS+6-BA1.0mg/L+NAA0.1mg/L of described strong seedling culture base.
Described root media comprises MS+6-IBA1.0-3.0mg/L+NAA0.05-0.2mg/L.
The preferred MS+6-IBA2.0mg/L+NAA0.2mg/L of described root media.
Described medium also comprises sucrose 20-40g/L, agar powder 2-4g/L, medium pH 5.5-6.0, cultivation temperature 22-28 ℃, illumination 1500-2500lx.
Compared with prior art, the present invention has greatly improved the reproduction speed of pink champagne clematis and the regularity of seedling, and has kept original maternal character better by tissue culture technology, can realize the batch production production in enormous quantities of growing seedlings.
Embodiment
The present invention is described in detail below in conjunction with specific embodiment.
Embodiment 1
(1) acquisition of sterilizable material
Get the bud on the young shoot of sprouting spring, with behind the running water flushing 1h on ultra-clean work team workbench, utilizing mass concentration successively is 60% alcohol immersion 10s, volumetric concentration is 0.5 ‰ mercury immersion 10min, aseptic water washing 4 times, after blotting surface moisture with aseptic filter paper, bud is cut into the sections of the long band axillalry bud of 0.5cm, is inoculated on the axillalry bud inducing culture that comprises MS+6-BA1.0mg/L+NAA0.05mg/L;
(2) differentiation of bud and propagation
The sections of band axillalry bud is inoculated on the axillalry bud inducing culture, axillalry bud position, 1 week back begins to expand, the green-yellow projection appears, the visible bud meristematic tissue in 3 week backs was cultivated 1 month again, and axillalry bud is long to 3cm, with axillalry bud cutting-out carrying out enrichment culture, the bud of differentiation changes in the adventitious bud proliferation medium that comprises 1.0mg/L MS+6-BA+NAA 0.05mg/L, though the base portion of the bud of differentiation has more callus, does not influence the propagation of indefinite bud;
(3) indefinite bud strong seedling culture
On the adventitious bud proliferation medium, in the bud of growing thickly that induces, every clump has 2 strains to extend, remaining is in the dwarfing state, after the bud of will growing thickly is divided into Xiao Cong, go on the strong seedling culture base that comprises MS+6-BA0.5mg/L+NAA0.1mg/L and grow, wherein the indefinite bud elongation can be grown to 3cm after 15 days rapidly;
(4) culture of rootage
Get the plantlet of 2cm, root induction in the root media of going into to comprise MS+6-IBA1.0mg/L+NAA0.05mg/L of transferring.The seedling base section dissolves the root original hase of white after 5 days, begins to take root after 18 days, can grow to 1cm after 30 days, and every strain has 3 roots, and rooting rate is 80%;
(5) hardening and transplanting
Culture of rootage 30 days when root system grows to 1cm, is selected the aseptic seedling of well developed root system, robust growth, indoor uncork hardening 1 day is taken out and is cleaned root agar, plants and tames after 20 days in the seedbed in greenhouse, can transplant basin, give rich water quality management, transplanting survival rate is 90%.
The medium of above-mentioned various situations also comprises sucrose 20g/L, agar powder 2g/L, pH=5.5,22 ℃ of cultivation temperature, illumination 1500lx.
Embodiment 2
(1) acquisition of sterilizable material
Get the bud on the young shoot of sprouting spring, with behind the running water flushing 2h on ultra-clean work team workbench, utilizing mass concentration successively is 75% alcohol immersion 10s, volumetric concentration is 1 ‰ mercury immersion 20min, aseptic water washing 5 times, after blotting surface moisture with aseptic filter paper, bud is cut into the sections of the long band axillalry bud of 1cm, is inoculated on the axillalry bud inducing culture that comprises MS+6-BA2.0mg/L+NAA0.2mg/L;
(2) differentiation of bud and propagation
The sections of band axillalry bud is inoculated on the axillalry bud inducing culture, axillalry bud position, 2 week back begins to expand, the green-yellow projection appears, the visible bud meristematic tissue in 3 week backs was cultivated 1 month again, and axillalry bud is long to 4cm, with axillalry bud cutting-out carrying out enrichment culture, the bud of differentiation changes in the adventitious bud proliferation medium that comprises MS+6-BA1.0mg/L+NAA0.2mg/L, though the base portion of the bud of differentiation has more callus, does not influence the propagation of indefinite bud;
(3) indefinite bud strong seedling culture
On the adventitious bud proliferation medium, in the bud of growing thickly that induces, every clump has 3 strains to extend, remaining is in the dwarfing state, after the bud of will growing thickly is divided into Xiao Cong, go on the strong seedling culture base that comprises MS+6-BA1mg/L+NAA0.1mg/L and grow, wherein the indefinite bud elongation can be grown to 4cm after 20 days rapidly;
(4) culture of rootage
Get the plantlet of 3cm, root induction in the root media of going into to comprise MS+6-IBA2.0mg/L+NAA0.2mg/L of transferring.The seedling base section dissolves the root original hase of white after 10 days, begins to take root after 20 days, can grow to 2cm after 30 days, and every strain has 4 roots, and rooting rate is 90%;
(5) hardening and transplanting
Culture of rootage 30 days when root system grows to 2cm, is selected the aseptic seedling of well developed root system, robust growth, indoor uncork hardening 2 days is taken out and is cleaned root agar, plants and tames after 30 days in the seedbed in greenhouse, can transplant basin, give rich water quality management, transplanting survival rate is 95%.
The medium of above-mentioned various situations also comprises sucrose 30g/L, agar powder 3g/L, pH=5.8,25 ℃ of cultivation temperature, illumination 2000lx.
Embodiment 3
(1) acquisition of sterilizable material
Get the bud on the young shoot of sprouting spring, with behind the running water flushing 3h on ultra-clean work team workbench, utilizing mass concentration successively is 75% alcohol immersion 50s, volumetric concentration is 2 ‰ mercury immersion 30min, aseptic water washing 6 times, after utilizing aseptic filter paper to blot surface moisture, bud is cut into the sections of the long band axillalry bud of 2cm, is inoculated on the axillalry bud inducing culture that comprises MS+6-BA3.0mg/L+NAA 0.2mg/L;
(2) differentiation of bud and propagation
The sections of band axillalry bud is inoculated on the axillalry bud inducing culture, axillalry bud position, 3 week back begins to expand, the green-yellow projection appears, the visible bud meristematic tissue in 5 week backs was cultivated 2 months again, and axillalry bud is long to 5cm, with axillalry bud cutting-out carrying out enrichment culture, the bud of differentiation changes in the adventitious bud proliferation medium that comprises MS+6-BA3.0mg/L+NAA0.2mg/L, though the base portion of the bud of differentiation has more callus, does not influence the propagation of indefinite bud;
(3) indefinite bud strong seedling culture
On the adventitious bud proliferation medium, in the bud of growing thickly that induces, every clump has 3 strains to extend, remaining is in the dwarfing state, after the bud of will growing thickly is divided into Xiao Cong, go on the strong seedling culture base that comprises MS+6-BA2mg/L+NAA0.2mg/L and grow, wherein the indefinite bud elongation can be grown to 5cm after 30 days rapidly;
(4) culture of rootage
Get the plantlet of 3cm, root induction in the root media of going into to comprise MS+6-IBA3.0mg/L+NAA0.2mg/L of transferring.The seedling base section dissolves the root original hase of white after 15 days, begins to take root after 25 days, can grow to 2cm after 35 days, and every strain has 4 roots, and rooting rate is 87%;
(5) hardening and transplanting
Culture of rootage 35 days when root system grows to 2cm, is selected the aseptic seedling of well developed root system, robust growth, indoor uncork hardening 3 days is taken out and is cleaned root agar, plants and tames after 40 days in the seedbed in greenhouse, can transplant basin, give rich water quality management, transplanting survival rate is 92%.
The medium of above-mentioned various situations also comprises sucrose 40g/L, agar powder 4g/L, pH=6.0,28 ℃ of cultivation temperature, illumination 2500lx.
Claims (10)
1. the method for tissue culture of pink champagne clematis is characterized in that, this method may further comprise the steps:
(1) acquisition of sterilizable material
Get the bud on the young shoot of sprouting spring, with behind the running water flushing 1-3h on ultra-clean work team workbench, utilize the alcohol immersion 10-50s of mass concentration successively for 60-75%, volumetric concentration is that the mercury of 0.5-2 ‰ soaks 10-30min, use aseptic water washing 4-6 time again, after utilizing aseptic filter paper to blot the moisture on bud surface, bud is cut into the sections of the long band axillalry bud of 0.5-2cm, is inoculated on the axillalry bud inducing culture;
(2) differentiation of bud and propagation
The sections of band axillalry bud is inoculated on the axillalry bud inducing culture, 1-3 began to expand in the axillalry bud position after week, the green-yellow projection appears, 3-5 is visible bud meristematic tissue after week, cultivates 1-2 month again, and axillalry bud is long to 3-5cm, with axillalry bud cutting-out carrying out enrichment culture, the bud of differentiation changes in the adventitious bud proliferation medium, though the base portion of the bud of differentiation has more callus, does not influence the propagation of indefinite bud;
(3) indefinite bud strong seedling culture
On the adventitious bud proliferation medium, every clump has the 2-4 strain to extend in the bud of growing thickly that induces, and remaining bud of growing thickly is in the dwarfing state, after the bud of will growing thickly is divided into Xiao Cong, go on the strong seedling culture base and grow, wherein the indefinite bud elongation can be grown to 3-5cm after 15-30 days rapidly;
(4) culture of rootage
Get the plantlet of 2-3cm, root induction in the root media is gone in switching, and the seedling base section dissolves the root original hase of white after 5-15 days, begins to take root after 18-25 days, can grow to 1-2cm after 30-35 days, and every strain has 3-4 root, and rooting rate is 80-90%;
(5) hardening and transplanting
Culture of rootage 30-35 days, when root system grows to 1-2cm, select the aseptic seedling of well developed root system, robust growth, in indoor uncork hardening 1-3 days, then seedling is taken out, clean the agar of root, plant in the seedbed in greenhouse and tamed 20-40 days, can transplant basin, give rich water quality management, final transplanting survival rate is 90-95%.
2. the method for tissue culture of pink champagne clematis according to claim 1 is characterized in that, described axillalry bud inducing culture comprises MS+6-BA1.0-3.0mg/L+NAA0.05-0.2mg/L.
3. the method for tissue culture of pink champagne clematis according to claim 2 is characterized in that, the preferred MS+6-BA2.0mg/L+NAA0.2mg/L of described axillalry bud inducing culture.
4. the method for tissue culture of pink champagne clematis according to claim 1 is characterized in that, described adventitious bud proliferation medium comprises MS+6-BA1.0-3.0mg/L+NAA0.05-0.2mg/L.
5. the method for tissue culture of pink champagne clematis according to claim 4 is characterized in that, the preferred MS+6-BA1.0mg/L+NAA0.2mg/L of described adventitious bud proliferation medium.
6. the method for tissue culture of pink champagne clematis according to claim 1 is characterized in that, described strong seedling culture base comprises MS+6-BA0.5-2.0mg/L+NAA0.1-0.2mg/L.
7. the method for tissue culture of pink champagne clematis according to claim 6 is characterized in that, the preferred MS+6-BA1.0mg/L+NAA0.1mg/L of described strong seedling culture base.
8. the method for tissue culture of pink champagne clematis according to claim 1 is characterized in that, described root media comprises MS+6-IBA1.0-3.0mg/L+NAA0.05-0.2mg/L.
9. the method for tissue culture of pink champagne clematis according to claim 8 is characterized in that, the preferred MS+6-IBA2.0mg/L+NAA0.2mg/L of described root media.
10. according to the method for tissue culture of claim 2 or 4 or 6 or 8 described pink champagne clematis, it is characterized in that described medium also comprises sucrose 20-40g/L, agar powder 2-4g/L, medium pH 5.5-6.0, cultivation temperature 22-28 ℃, illumination 1500-2500lx.
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102119663A (en) * | 2011-01-05 | 2011-07-13 | 南京林业大学 | Tissue culture quick propagation technology of clematis mademe Julia correvon |
| CN102144543A (en) * | 2011-01-05 | 2011-08-10 | 南京林业大学 | Tissue culture and rapid propagation technology for Clematis 'Arabella' |
| 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 |
| 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 |
| 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 |
| CN108207621A (en) * | 2016-12-21 | 2018-06-29 | 中国科学院上海生命科学研究院 | A kind of method that direct adventitious bud of Actions of Clematis Species occurs in vitro |
| CN110651713A (en) * | 2019-10-29 | 2020-01-07 | 上海植物园 | Tissue culture method of clematis' Fuji blue |
| 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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2009
- 2009-04-24 CN CN200910049978A patent/CN101869070A/en active Pending
Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102119663A (en) * | 2011-01-05 | 2011-07-13 | 南京林业大学 | Tissue culture quick propagation technology of clematis mademe Julia correvon |
| CN102144543A (en) * | 2011-01-05 | 2011-08-10 | 南京林业大学 | Tissue culture and rapid propagation technology for Clematis 'Arabella' |
| 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 |
| 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 |
| CN104082151A (en) * | 2014-08-04 | 2014-10-08 | 云南农业大学 | Cultivation method for polyploid clematis ranunculoides |
| CN104082152A (en) * | 2014-08-04 | 2014-10-08 | 云南农业大学 | Tissue culture and rapid propagation method for clematis ranunculoides |
| 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 |
| CN108207621A (en) * | 2016-12-21 | 2018-06-29 | 中国科学院上海生命科学研究院 | A kind of method that direct adventitious bud of Actions of Clematis Species occurs in vitro |
| CN110651713A (en) * | 2019-10-29 | 2020-01-07 | 上海植物园 | Tissue culture method of clematis' Fuji blue |
| CN110651713B (en) * | 2019-10-29 | 2022-09-13 | 上海植物园 | Tissue culture method of clematis' Fuji blue |
| 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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Open date: 20101027 |