CN101869067A - Tissue culture method for anemone - Google Patents
Tissue culture method for anemone Download PDFInfo
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- CN101869067A CN101869067A CN200910049975A CN200910049975A CN101869067A CN 101869067 A CN101869067 A CN 101869067A CN 200910049975 A CN200910049975 A CN 200910049975A CN 200910049975 A CN200910049975 A CN 200910049975A CN 101869067 A CN101869067 A CN 101869067A
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Abstract
The invention relates to a tissue culture method for anemone, which comprises the following steps of: acquiring a sterile material, differentiating and proliferating buds, culturing adventitious buds and strong seedlings, performing root culture, hardening and transplanting seedlings, and the like. Compared with the prior art, the method has the advantages of greatly improving the reproduction speed of the anemone and the uniformity of the seedlings, improving the stability of characters of the anemone, and realizing industrial mass production of seedling culture.
Description
Technical field
The present invention relates to the method for tissue culture of a plant species, especially relate to the method for tissue culture of anemone.
Background technology
Anemone is Ranunculaceae, thimbleweed, is widely distributed in all over the world, is most commonly in the forest land and the grassy marshland of north temperate zone, is perennial herb.Anemone basal leaf 4-8 sheet, the long 6-30cm of petiole dredges the growth pubescence.Blade is round kidney shape, and 3 totally cleave, 2-5 in flower, diameter 3.5-5cm, pink.The florescence of anemone is autumn, and the long draft hair is arranged on the achene.Because of pattern is bright-coloured, anemone extensively cultivation in the garden, be welcome fall flowering along ray flower altar flowers, still, as the new varieties of external introduction, anemone introduce a fine variety negligible amounts, the seedling supply is subjected to certain restriction.
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 improves the method for tissue culture of the anemone of property stability.
Purpose of the present invention can be achieved through the following technical solutions:
The method for tissue culture of anemone is characterized in that, this method may further comprise the steps:
(1) acquisition of sterilizable material
Get the young tender bud of sprouting, after removing the blade of coated outside, with behind the running water flushing 1-3h on superclean bench, utilize the alcohol immersion 10-50s of mass concentration successively for 70-75%, volumetric concentration is that the mercury of 0.5-2 ‰ soaks 10-30min, uses aseptic water washing 4-6 time again, utilize aseptic filter paper to blot surperficial moisture after, be cut into the sections of the long band axillalry bud of 0.5-2cm again, sections is inoculated on the bud inducing culture;
(2) differentiation of bud and propagation
Sections is inoculated on the bud inducing culture 2-4 after week, the axillalry bud position begins to expand, the yellow green projection occurs, 3-5 is visible bud meristematic tissue after week, cultivates 1-2 month again, indefinite bud can be grown 2-3cm, indefinite bud downcut in the proliferated culture medium change bud over to carry out enrichment culture, the callus of indefinite bud base portion is more, but does not influence propagation, indefinite bud growth rapidly and do not have bad phenomenon such as vitrifying, it is good to grow in medium;
(3) indefinite bud strong seedling culture
On the proliferated culture medium of bud, every clump has the 2-3 strain to extend in the bud of growing thickly that induces, and remaining is in the dwarfing state, after the bud of will growing thickly is divided into Xiao Cong, goes on the strong seedling culture base and grows, and indefinite bud extends rapidly;
(4) culture of rootage
Get the indefinite bud plantlet of 3-4cm, 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, can grow to 4-6cm after 25-40 days, and root system is sturdy, and fibrous root is numerous, and rooting rate is 90-100%;
(5) hardening and transplanting
Culture of rootage 15-30 days, when root system grows to 0.5-1cm, select the aseptic seedling of well developed root system, robust growth, in indoor uncork hardening 2-4 days, then seedling is taken out, clean the agar of root, plant in the seedbed in the greenhouse and tamed 20-40 days, can transplant outdoorly, give rich water quality management, final transplanting survival rate is 90-95%.
Described bud inducing culture comprises MS+6-BA1.0-5.0mg/L+NAA0.1-0.5mg/L.
The preferred MS+6-BA3.0mg/L+NAA0.3mg/L of described bud inducing culture.
The proliferated culture medium of described bud comprises MS+6-BA0.5-2.0mg/L+NAA0.1-0.2mg/L.
The preferred MS+6-BA2.0mg/L+NAA0.2mg/L of the proliferated culture medium of described bud.
Described strong seedling culture base comprises MS+6-BA0.5-2.0mg/L+NAA0.1-0.2mg/L.
The preferred MS+6-BA0.5mg/L+NAA0.1mg/L of described strong seedling culture base.
Described root media comprises MS+NAA0.05-0.2mg/L.
The preferred MS+NAA0.05mg/L of described root media.
Described medium also comprises sucrose 20-40g/L, agar powder 4-8g/L, medium pH 5.5-6.0, cultivation temperature 24-26 ℃, illumination 1500-2500lx.
Compared with prior art, the present invention has greatly improved the reproduction speed of anemone and the regularity of seedling, and has improved the stability of its proterties 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 young tender bud of sprouting, after removing the blade of coated outside, with behind the running water flushing 1h on superclean bench, utilizing mass concentration successively is 70% alcohol immersion 10s, volumetric concentration is that 0.5 ‰ mercury soaks 10min, uses aseptic water washing again 4 times, utilize aseptic filter paper to blot the moisture on surface after, be cut into the sections of the long band axillalry bud of 0.5cm again, sections is inoculated on the bud inducing culture that comprises MS+6-BA1.0mg/L+NAA0.1mg/L;
(2) differentiation of bud and propagation
Sections was inoculated on the bud inducing culture after 2 weeks, the axillalry bud position begins to expand, the yellow green projection occurs, the visible bud meristematic tissue in 3 week backs was cultivated 1 month again, indefinite bud can be grown 2cm, indefinite bud downcut in the proliferated culture medium change the bud that comprises MS+6-BA0.5mg/L+NAA0.1mg/L over to carry out enrichment culture, the callus of indefinite bud base portion is more, but does not influence propagation, indefinite bud growth rapidly and do not have bad phenomenon such as vitrifying, it is good to grow in medium;
(3) indefinite bud strong seedling culture
On the proliferated culture medium of bud, every clump has 2 strains to extend in the bud of growing thickly that induces, and remaining is in the dwarfing state, after the bud of will growing thickly is divided into Xiao Cong, goes on the strong seedling culture base that comprises MS+6-BA0.5mg/L+NAA0.1mg/L and grows, and indefinite bud extends rapidly;
(4) culture of rootage
Get the indefinite bud plantlet of 3cm, root induction in the root media of going into to comprise MS+NAA0.05mg/L of transferring, the seedling base section dissolves the root original hase of white after 5 days, can grow to 4cm after 25 days, and root system is sturdy, and fibrous root is numerous, and rooting rate is 90%;
(5) hardening and transplanting
Culture of rootage 15 days, when root system grows to 0.5cm, select the aseptic seedling of well developed root system, robust growth, in indoor uncork hardening 2 days, then seedling is taken out, clean the agar of root, plant in the seedbed in the greenhouse and tamed 20 days, can transplant outdoorly, give rich water quality management, final transplanting survival rate is 90%.
The medium of above-mentioned various situations also comprises sucrose 20g/L, agar powder 4g/L, pH=5.5,24 ℃ of cultivation temperature, illumination 1500lx.
Embodiment 2
(1) acquisition of sterilizable material
Get the young tender bud of sprouting, after removing the blade of coated outside, with behind the running water flushing 2h on superclean bench, utilizing mass concentration successively is 75% alcohol immersion 30s, volumetric concentration is that 1 ‰ mercury soaks 15min, uses aseptic water washing again 5 times, utilize aseptic filter paper to blot the moisture on surface after, be cut into the sections of the long band axillalry bud of 1cm again, sections is inoculated on the bud inducing culture that comprises MS+6-BA3.0mg/L+NAA0.3mg/L;
(2) differentiation of bud and propagation
Sections was inoculated on the bud inducing culture after 3 weeks, the axillalry bud position begins to expand, the yellow green projection occurs, the visible bud meristematic tissue in 4 week backs is cultivated 1 first quarter moon again, indefinite bud can be grown 3cm, indefinite bud downcut in the proliferated culture medium change the bud that comprises MS+6-BA2.0mg/L+NAA0.2mg/L over to carry out enrichment culture, the callus of indefinite bud base portion is more, but does not influence propagation, indefinite bud growth rapidly and do not have bad phenomenon such as vitrifying, it is good to grow in medium;
(3) indefinite bud strong seedling culture
On the proliferated culture medium of bud, every clump has 3 strains to extend in the bud of growing thickly that induces, and remaining is in the dwarfing state, after the bud of will growing thickly is divided into Xiao Cong, goes on the strong seedling culture base that comprises MS+6-BA2.0mg/L+NAA0.2mg/L and grows, and indefinite bud extends rapidly;
(4) culture of rootage
Get the indefinite bud plantlet of 4cm, root induction in the root media of going into to comprise MS+NAA0.05mg/L of transferring, the seedling base section dissolves the root original hase of white after 10 days, can grow to 6cm after 30 days, and root system is sturdy, and fibrous root is numerous, and rooting rate is 100%;
(5) hardening and transplanting
Culture of rootage 20 days, when root system grows to 1cm, select the aseptic seedling of well developed root system, robust growth, in indoor uncork hardening 3 days, then seedling is taken out, clean the agar of root, plant in the seedbed in the greenhouse and tamed 30 days, can transplant outdoorly, give rich water quality management, final transplanting survival rate is 100%.
The medium of above-mentioned various situations also comprises sucrose 30g/L, agar powder 6g/L, pH=5.8,25 ℃ of cultivation temperature, illumination 2000lx.
Embodiment 3
(1) acquisition of sterilizable material
Get the young tender bud of sprouting, after removing the blade of coated outside, with behind the running water flushing 3h on superclean bench, utilizing mass concentration successively is 75% alcohol immersion 50s, volumetric concentration is that 2 ‰ mercury soaks 30min, uses aseptic water washing again 6 times, utilize aseptic filter paper to blot the moisture on surface after, be cut into the sections of the long band axillalry bud of 2cm again, sections is inoculated on the bud inducing culture that comprises MS+6-BA5.0mg/L+NAA0.5mg/L;
(2) differentiation of bud and propagation
Sections was inoculated on the bud inducing culture after 4 weeks, the axillalry bud position begins to expand, the yellow green projection occurs, the visible bud meristematic tissue in 5 week backs was cultivated 2 months again, indefinite bud can be grown 2cm, indefinite bud downcut in the proliferated culture medium change the bud that comprises MS+6-BA2.0mg/L+NAA0.2mg/L over to carry out enrichment culture, the callus of indefinite bud base portion is more, but does not influence propagation, indefinite bud growth rapidly and do not have bad phenomenon such as vitrifying, it is good to grow in medium;
(3) indefinite bud strong seedling culture
On the proliferated culture medium of bud, every clump has 3 strains to extend in the bud of growing thickly that induces, and remaining is in the dwarfing state, after the bud of will growing thickly is divided into Xiao Cong, goes on the strong seedling culture base that comprises MS+6-BA2.0mg/L+NAA0.2mg/L and grows, and indefinite bud extends rapidly;
(4) culture of rootage
Get the indefinite bud plantlet of 3cm, root induction in the root media of going into to comprise MS+NAA0.2mg/L of transferring, the seedling base section dissolves the root original hase of white after 15 days, can grow to 5cm after 40 days, and root system is sturdy, and fibrous root is numerous, and rooting rate is 96%;
(5) hardening and transplanting
Culture of rootage 30 days, when root system grows to 1cm, select the aseptic seedling of well developed root system, robust growth, in indoor uncork hardening 4 days, then seedling is taken out, clean the agar of root, plant in the seedbed in the greenhouse and tamed 40 days, can transplant outdoorly, give rich water quality management, final transplanting survival rate is 96%.
The medium of above-mentioned various situations also comprises sucrose 40g/L, agar powder 8g/L, pH=6.0,26 ℃ of cultivation temperature, illumination 2500lx.
Claims (10)
1. the method for tissue culture of anemone is characterized in that, this method may further comprise the steps:
(1) acquisition of sterilizable material
Get the young tender bud of sprouting, after removing the blade of coated outside, with behind the running water flushing 1-3h on superclean bench, utilize the alcohol immersion 10-50s of mass concentration successively for 70-75%, volumetric concentration is that the mercury of 0.5-2 ‰ soaks 10-30min, uses aseptic water washing 4-6 time again, utilize aseptic filter paper to blot surperficial moisture after, be cut into the sections of the long band axillalry bud of 0.5-2cm again, sections is inoculated on the bud inducing culture;
(2) differentiation of bud and propagation
Sections is inoculated on the bud inducing culture 2-4 after week, the axillalry bud position begins to expand, the yellow green projection occurs, 3-5 is visible bud meristematic tissue after week, cultivates 1-2 month again, indefinite bud can be grown 2-3cm, indefinite bud downcut in the proliferated culture medium change bud over to carry out enrichment culture, the callus of indefinite bud base portion is more, but does not influence propagation, indefinite bud growth rapidly and do not have bad phenomenon such as vitrifying, it is good to grow in medium;
(3) indefinite bud strong seedling culture
On the proliferated culture medium of bud, every clump has the 2-3 strain to extend in the bud of growing thickly that induces, and remaining is in the dwarfing state, after the bud of will growing thickly is divided into Xiao Cong, goes on the strong seedling culture base and grows, and indefinite bud extends rapidly;
(4) culture of rootage
Get the indefinite bud plantlet of 3-4cm, 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, can grow to 4-6cm after 25-40 days, and root system is sturdy, and fibrous root is numerous, and rooting rate is 90-100%;
(5) hardening and transplanting
Culture of rootage 15-30 days, when root system grows to 0.5-1cm, select the aseptic seedling of well developed root system, robust growth, in indoor uncork hardening 2-4 days, then seedling is taken out, clean the agar of root, plant in the seedbed in the greenhouse and tamed 20-40 days, can transplant outdoorly, give rich water quality management, final transplanting survival rate is 90-95%.
2. the method for tissue culture of anemone according to claim 1 is characterized in that, described bud inducing culture comprises MS+6-BA1.0-5.0mg/L+NAA0.1-0.5mg/L.
3. the method for tissue culture of anemone according to claim 2 is characterized in that, the preferred MS+6-BA3.0mg/L+NAA0.3mg/L of described bud inducing culture.
4. the method for tissue culture of anemone according to claim 1 is characterized in that, the proliferated culture medium of described bud comprises MS+6-BA0.5-2.0mg/L+NAA0.1-0.2mg/L.
5. the method for tissue culture of anemone according to claim 4 is characterized in that, the preferred MS+6-BA2.0mg/L+NAA0.2mg/L of the proliferated culture medium of described bud.
6. the method for tissue culture of anemone 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 anemone according to claim 6 is characterized in that, the preferred MS+6-BA0.5mg/L+NAA0.1mg/L of described strong seedling culture base.
8. the method for tissue culture of anemone according to claim 1 is characterized in that, described root media comprises MS+NAA0.05-0.2mg/L.
9. the method for tissue culture of anemone according to claim 8 is characterized in that, the preferred MS+NAA0.05mg/L of described root media.
10. according to the method for tissue culture of claim 2 or 4 or 6 or 8 described anemones, it is characterized in that described medium also comprises sucrose 20-40g/L, agar powder 4-8g/L, medium pH 5.5-6.0, cultivation temperature 24-26 ℃, illumination 1500-2500lx.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009100499752A CN101869067B (en) | 2009-04-24 | 2009-04-24 | Tissue culture method for anemone |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009100499752A CN101869067B (en) | 2009-04-24 | 2009-04-24 | Tissue culture method for anemone |
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| Publication Number | Publication Date |
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| CN101869067A true CN101869067A (en) | 2010-10-27 |
| CN101869067B CN101869067B (en) | 2012-01-04 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN2009100499752A Expired - Fee Related CN101869067B (en) | 2009-04-24 | 2009-04-24 | Tissue culture method for anemone |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104396749A (en) * | 2014-11-18 | 2015-03-11 | 罗翼 | Tissue culture propagation method of Anemone vitifolia |
| CN104396748A (en) * | 2014-11-18 | 2015-03-11 | 罗翼 | Tissue culture propagation method of anemone dichotoma |
| CN113951138A (en) * | 2021-11-08 | 2022-01-21 | 河北农业大学 | Rapid propagation method of anemone macrocephala |
-
2009
- 2009-04-24 CN CN2009100499752A patent/CN101869067B/en not_active Expired - Fee Related
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104396749A (en) * | 2014-11-18 | 2015-03-11 | 罗翼 | Tissue culture propagation method of Anemone vitifolia |
| CN104396748A (en) * | 2014-11-18 | 2015-03-11 | 罗翼 | Tissue culture propagation method of anemone dichotoma |
| CN113951138A (en) * | 2021-11-08 | 2022-01-21 | 河北农业大学 | Rapid propagation method of anemone macrocephala |
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| Publication number | Publication date |
|---|---|
| CN101869067B (en) | 2012-01-04 |
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Owner name: STATE GRID SHANGHAI ELECTRIC POWER COMPANY POWER S Effective date: 20140107 Owner name: SHANGHAI SHANGFANG LANDSCAPE PLANT RESEARCH INSTIT Free format text: FORMER OWNER: SHANGHAI SHANGFANG LANDSCAPE PLANT INSTITUTE Effective date: 20140107 |
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Effective date of registration: 20140107 Address after: 201114 Pujiang village, Pujiang Town, Shanghai, Minhang District Patentee after: SHANGHAI SHANGFANG GARDEN PLANT RESEARCH INSTITUTE CO., LTD. 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: Shanghai Shangfang Landscape Plant Institute |
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