CN101869068B - Tissue culture method for tulbaghia violacea - Google Patents
Tissue culture method for tulbaghia violacea Download PDFInfo
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- CN101869068B CN101869068B CN2009100499767A CN200910049976A CN101869068B CN 101869068 B CN101869068 B CN 101869068B CN 2009100499767 A CN2009100499767 A CN 2009100499767A CN 200910049976 A CN200910049976 A CN 200910049976A CN 101869068 B CN101869068 B CN 101869068B
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Abstract
The invention relates to a tissue culture method for tulbaghia violacea. The method comprises the following steps of: obtaining sterile materials; differentiating and proliferating buds; culturing strong seedlings of adventitious buds; rooting; and hardening off seedlings and transplanting the seedlings and the like. Compared with the prior art, the method greatly improves the reproductive speed of the tulbaghia violacea and the trimming of the seedlings, and improves the stability of shape and properties of the tulbaghia violacea, so that industrial mass production of the seedlings can be realized.
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 tulbaghia violacea.
Background technology
Tulbaghia violacea is the Liliaceae allium, and the original producton location is South Africa, belongs to flowering bulb, plant height 30-50cm, and the cylindrical clove of tool becomes the strain shape of growing thickly.Cauline leaf all contains the fragrant-flowered garlic flavor, and the sympodium inflorescence is arranged at the top, opens purple pink colour Xiao Hua, and the florescence is long, is that good garden is planted, potted plant or cut-flower material.But as the new varieties of external introduction, tulbaghia violacea is introduced a fine variety negligible amounts, and the plantation of its plant division or bulb is slow, so the seedling supply receives certain restriction, can't satisfy the demand in present market.
Summary of the invention
The object of the invention is exactly for the defective that overcomes above-mentioned prior art existence a kind of regularity that improves reproduction speed and seedling to be provided, and improves the method for tissue culture of the tulbaghia violacea of property stability.
The object of the invention can be realized through following technical scheme:
The method for tissue culture of tulbaghia violacea is characterized in that, this method may further comprise the steps:
(1) acquisition of sterilizable material
Get the young tender budlet and the petal of sprouting spring; After removing blade; 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 and the petal of the long band axillalry bud of 0.5-2cm again, sections and petal are inoculated on the bud inducing culture;
(2) differentiation of bud and propagation
Sections and petal are inoculated on the bud inducing culture 2-4 after week; The axillalry bud position begins to expand; The yellow green projection occurs, 4-6 is visible bud separate living tissue after week, cultivates 1-3 month again; Little adventitious bud can be grown 2-3cm; Adventitious bud downcut in the proliferated culture medium change bud over to breed cultivation, the callus of adventitious bud base portion is more, but does not influence propagation; Adventitious bud growth rapidly and do not have bad phenomenon such as vitrifying, it is good in culture medium, to grow;
(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, can grow to 3-4cm after 15-30 days;
(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 10-20 days, can grow to 3-4cm after 25-40 days, and root system is sturdy, and fibrous root is numerous, and rooting rate is 90-100%;
(5) refining seedling and transplanting
Culture of rootage 20-40 days, when root system grows to 0.5-1cm, select the aseptic seedling of well developed root system, robust growth; Refine seedling 1-4 days in indoor uncork, 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 90-100%.
Described bud inducing culture comprises MS+6-BA2.0-5.0mg/L+NAA0.2-0.5mg/L.
The preferred MS+6-BA5.0mg/L+NAA0.5mg/L of described bud inducing culture.
The proliferated culture medium of described bud comprises MS+6-BA1.0-3.0mg/L+NAA0.1-0.3mg/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-BA1.0mg/L+NAA0.1mg/L of described strong seedling culture base.
Described root media comprises MS+NAA0.1-0.3mg/L.
The preferred MS+NAA0.2mg/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 tulbaghia violacea and the regularity of seedling, and has improved the stability of its proterties through tissue culture technology, can realize the batch production production in enormous quantities of growing seedlings.
Embodiment
Below in conjunction with specific embodiment the present invention is elaborated.
Embodiment 1
(1) acquisition of sterilizable material
Get the young tender budlet and the petal of sprouting spring; After removing blade; 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 and the petal of the long band axillalry bud of 0.5cm again, sections and petal are inoculated on the bud inducing culture that comprises MS+6-BA2.0mg/L+NAA0.2mg/L;
(2) differentiation of bud and propagation
Sections and petal were inoculated on the bud inducing culture after 2 weeks; The axillalry bud position begins to expand; The yellow green projection occurs, the visible bud separate living tissue in 4 week backs was cultivated 1 month again; Little adventitious bud can be grown 2cm; Adventitious bud downcut in the proliferated culture medium change the bud that comprises MS+6-BA1.0mg/L+NAA0.1mg/L over to breed cultivation, the callus of adventitious bud base portion is more, but does not influence propagation; Adventitious bud growth rapidly and do not have bad phenomenon such as vitrifying, it is good in culture medium, to grow;
(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; Go on the strong seedling culture base that comprises MS+6-BA0.5mg/L+NAA0.1mg/L and grow, indefinite bud extends rapidly, can grow to 3cm after 15 days;
(4) culture of rootage
Get the indefinite bud plantlet of 3cm, root induction in the root media of going into to comprise MS+NAA0.1mg/L of transferring, the seedling base section dissolves the root original hase of white after 10 days, can grow to 3cm after 25 days, and root system is sturdy, and fibrous root is numerous, and rooting rate is 90%;
(5) refining seedling and transplanting
Culture of rootage 20 days when root system grows to 0.5cm, is selected the aseptic seedling of well developed root system, robust growth; Refine seedling 1 day in indoor uncork, 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 situation 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 budlet and the petal of sprouting spring; After removing blade; 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 and the petal of the long band axillalry bud of 1cm again, sections and petal are inoculated on the bud inducing culture that comprises MS+6-BA5.0mg/L+NAA0.5mg/L;
(2) differentiation of bud and propagation
Sections and petal were inoculated on the bud inducing culture after 3 weeks; The axillalry bud position begins to expand; The yellow green projection occurs, the visible bud separate living tissue in 5 week backs was cultivated 2 months again; Little adventitious bud can be grown 3cm; Adventitious bud downcut in the proliferated culture medium change the bud that comprises MS+6-BA2.0mg/L+NAA0.2mg/L over to breed cultivation, the callus of adventitious bud base portion is more, but does not influence propagation; Adventitious bud growth rapidly and do not have bad phenomenon such as vitrifying, it is good in culture medium, to grow;
(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; Go on the strong seedling culture base that comprises MS+6-BA1.0mg/L+NAA0.1mg/L and grow, indefinite bud extends rapidly, can grow to 4cm after 20 days;
(4) culture of rootage
Get the indefinite bud plantlet of 4cm, 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 4cm after 30 days, and root system is sturdy, and fibrous root is numerous, and rooting rate is 100%;
(5) refining seedling 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; Refine seedling 3 days in indoor uncork, 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 situation 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 budlet and the petal of sprouting spring; After removing blade; 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 and the petal of the long band axillalry bud of 2cm again, sections and petal are inoculated on the bud inducing culture that comprises MS+6-BA5.0mg/L+NAA0.5mg/L;
(2) differentiation of bud and propagation
Sections and petal were inoculated on the bud inducing culture after 4 weeks; The axillalry bud position begins to expand; The yellow green projection occurs, the visible bud separate living tissue in 6 week backs was cultivated 2 months again; Little adventitious bud can be grown 3cm; Adventitious bud downcut in the proliferated culture medium change the bud that comprises MS+6-BA3.0mg/L+NAA0.3mg/L over to breed cultivation, the callus of adventitious bud base portion is more, but does not influence propagation; Adventitious bud growth rapidly and do not have bad phenomenon such as vitrifying, it is good in culture medium, to grow;
(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; Go on the strong seedling culture base that comprises MS+6-BA2.0mg/L+NAA0.2mg/L and grow, indefinite bud extends rapidly, can grow to 4cm after 30 days;
(4) culture of rootage
Get the indefinite bud plantlet of 4cm, root induction in the root media of going into to comprise MS+NAA0.3mg/L of transferring, the seedling base section dissolves the root original hase of white after 20 days, can grow to 4cm after 40 days, and root system is sturdy, and fibrous root is numerous, and rooting rate is 94%;
(5) refining seedling and transplanting
Culture of rootage 40 days when root system grows to 1cm, is selected the aseptic seedling of well developed root system, robust growth; Refine seedling 4 days in indoor uncork, 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 95%.
The medium of above-mentioned various situation also comprises sucrose 40g/L, agar powder 8g/L, pH=6.0,26 ℃ of cultivation temperature, illumination 2500lx.
Claims (6)
1. the method for tissue culture of tulbaghia violacea is characterized in that, this method may further comprise the steps:
(1) acquisition of sterilizable material
Get the young tender budlet and the petal of sprouting spring; After removing blade; 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 and the petal of the long band axillalry bud of 0.5-2cm again, sections and petal are inoculated on the bud inducing culture;
(2) differentiation of bud and propagation
Sections and petal are inoculated on the bud inducing culture 2-4 after week; The axillalry bud position begins to expand; The yellow green projection occurs, 4-6 sees the bud separate living tissue after week, cultivates 1-3 month again; Little adventitious bud is long to 2-3cm; Adventitious bud downcut in the proliferated culture medium change bud over to breed cultivation, the callus of adventitious bud base portion is more, but does not influence propagation; The adventitious bud growth is rapid and do not have the vitrifying bad phenomenon, and it is good in culture medium, to grow;
(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, grows to 3-4cm after 15-30 days;
(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 10-20 days, grows to 3-4cm after 25-40 days, and root system is sturdy, and fibrous root is numerous, and rooting rate is 90-100%;
(5) refining seedling and transplanting
Culture of rootage 20-40 days; When root system grows to 0.5-1cm; Select the aseptic seedling of well developed root system, robust growth; Refine seedling 1-4 days in indoor uncork, then seedling is taken out, clean the agar of root; Plant in the seedbed in the greenhouse and tamed 20-40 days; Promptly transplant outdoorly, give rich water quality management, final transplanting survival rate is 90-100%;
Described bud inducing culture comprises MS+6-BA2.0-5.0mg/L+NAA0.2-0.5mg/L;
The proliferated culture medium of described bud comprises MS+6-BA1.0-3.0mg/L+NAA0.1-0.3mg/L;
Described strong seedling culture base comprises MS+6-BA0.5-2.0mg/L+NAA0.1-0.2mg/L;
Described root media comprises MS+NAA0.1-0.3mg/L.
2. the method for tissue culture of tulbaghia violacea according to claim 1 is characterized in that, described bud inducing culture comprises MS+6-BA5.0mg/L+NAA0.5mg/L.
3. the method for tissue culture of tulbaghia violacea according to claim 1 is characterized in that, the proliferated culture medium of described bud comprises MS+6-BA2.0mg/L+NAA0.2mg/L.
4. the method for tissue culture of tulbaghia violacea according to claim 1 is characterized in that, described strong seedling culture base comprises MS+6-BA1.0mg/L+NAA0.1mg/L.
5. the method for tissue culture of tulbaghia violacea according to claim 1 is characterized in that, described root media comprises MS+NAA0.2mg/L.
6. the method for tissue culture of tulbaghia violacea according to claim 1 is characterized in that, each 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 respectively.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104488709A (en) * | 2014-12-04 | 2015-04-08 | 浙江省农业科学院 | Method for culturing bulb tissues of tulbaghia violacea floral leaf |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102144549B (en) * | 2011-01-19 | 2013-12-25 | 宁波城市职业技术学院 | Tissue culture method of tulbaghia violacea harvey bud |
| CN103749304B (en) * | 2014-01-20 | 2016-06-01 | 上海上房园艺有限公司 | The tissue culture method of floral leaf mountain villous themeda orchid |
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Non-Patent Citations (2)
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| 张连全.夏日的清凉花紫娇花.《园林》.2006,(第10期),41. * |
| 杨勇,等.薤白愈伤组织诱导及其对草甘膦抗性鉴定.《湖南农业大学学报(自然科学版)》.2007,第33卷(第3期),277-280. * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104488709A (en) * | 2014-12-04 | 2015-04-08 | 浙江省农业科学院 | Method for culturing bulb tissues of tulbaghia violacea floral leaf |
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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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