CN101869053B - Tissue culture method of floral leaf pampasgrass - Google Patents
Tissue culture method of floral leaf pampasgrass Download PDFInfo
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- CN101869053B CN101869053B CN2009100498548A CN200910049854A CN101869053B CN 101869053 B CN101869053 B CN 101869053B CN 2009100498548 A CN2009100498548 A CN 2009100498548A CN 200910049854 A CN200910049854 A CN 200910049854A CN 101869053 B CN101869053 B CN 101869053B
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- 241000722863 Cortaderia jubata Species 0.000 title claims abstract description 17
- 238000012136 culture method Methods 0.000 title abstract 2
- 239000000463 material Substances 0.000 claims abstract description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 15
- 230000001939 inductive effect Effects 0.000 claims description 14
- 230000035755 proliferation Effects 0.000 claims description 14
- 238000000034 method Methods 0.000 claims description 13
- 229920001817 Agar Polymers 0.000 claims description 10
- 239000008272 agar Substances 0.000 claims description 10
- 241000196324 Embryophyta Species 0.000 claims description 8
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 5
- 206010020649 Hyperkeratosis Diseases 0.000 claims description 5
- 241000283986 Lepus Species 0.000 claims description 5
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 5
- 229930006000 Sucrose Natural products 0.000 claims description 5
- 230000028446 budding cell bud growth Effects 0.000 claims description 5
- 230000004069 differentiation Effects 0.000 claims description 5
- 238000011010 flushing procedure Methods 0.000 claims description 5
- 230000012010 growth Effects 0.000 claims description 5
- 238000005286 illumination Methods 0.000 claims description 5
- 238000007654 immersion Methods 0.000 claims description 5
- 230000006698 induction Effects 0.000 claims description 5
- QSHDDOUJBYECFT-UHFFFAOYSA-N mercury Chemical compound [Hg] QSHDDOUJBYECFT-UHFFFAOYSA-N 0.000 claims description 5
- 229910052753 mercury Inorganic materials 0.000 claims description 5
- 230000000442 meristematic effect Effects 0.000 claims description 5
- 239000000843 powder Substances 0.000 claims description 5
- 238000007670 refining Methods 0.000 claims description 5
- 239000005720 sucrose Substances 0.000 claims description 5
- 230000004083 survival effect Effects 0.000 claims description 5
- 238000005406 washing Methods 0.000 claims description 5
- 238000010923 batch production Methods 0.000 abstract description 2
- 238000009395 breeding Methods 0.000 abstract 2
- 230000001488 breeding effect Effects 0.000 abstract 2
- 238000005516 engineering process Methods 0.000 description 2
- 244000025254 Cannabis sativa Species 0.000 description 1
- 235000014676 Phragmites communis Nutrition 0.000 description 1
- 241001529246 Platymiscium Species 0.000 description 1
- 241000209504 Poaceae Species 0.000 description 1
- 240000001398 Typha domingensis Species 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
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- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention relates to a tissue culture method of floral leaf pampasgrass, comprising the steps of obtaining sterile materials, splitting and breeding germ, cultivating adventitious bud strong seedling, cultivating root, hardening seedling, replanting, and the like. Compared with the prior art, the invention greatly improves the breeding speed of floral leaf pampasgrass and the uniformity of the seedling, improves the stability of the character and can realize factory and batch production of grow seedlings.
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 floral leaf pampasgrass.
Background technology
Floral leaf pampasgrass is a grass family cattail and reed platymiscium, distributes extensively, and be evergreen herbaceos perennial, plant height 80-100cm, leaf adularescent striped, the conical featheriness of inflorescence is silvery white in color, and is the external famous grass of viewing and admiring, and is used for afforestation or bank more.The leaf of floral leaf pampasgrass, flower fringe are beautiful, and the garden cultivation is grand and graceful, plants in the bank, and autumn has set in can appreciate the panicle of its silvery white pinniform fringe, also can be used as dried flower or flower border to view and admire use careless garden belonging to a category in, has good annidation and ornamental value.But as the external new varieties of introducing, floral leaf pampasgrass is introduced a fine variety negligible amounts, and the seedling supply receives certain restriction.
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 floral leaf pampasgrass of property stability.
The object of the invention can be realized through following technical scheme:
The method for tissue culture of floral leaf pampasgrass is characterized in that, this method may further comprise the steps:
(1) acquisition of sterilizable material
Get the young tender bud of sprouting spring, remove the blade of coated outside after, 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 axillalry bud inducing culture;
(2) differentiation of bud and propagation
Sections is inoculated in that 1-3 is after week on the axillalry bud inducing culture, and the axillalry bud position begins to expand, and green projection occurs; 3-5 is visible bud meristematic tissue after week, cultivates 1-2 month again, and little indefinite bud can be grown 3-4cm; Indefinite bud downcut to change over to carry out enrichment culture in the adventitious bud proliferation medium, 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 in medium, to grow;
(3) indefinite bud strong seedling culture
On the adventitious bud proliferation medium, 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 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 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) refining seedling 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; Refine seedling 2-4 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 axillalry bud inducing culture comprises MS+6-BA1.0-5.0mg/L+NAA0.1-0.5mg/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-BA0.5-4.0mg/L+NAA0.1-0.2mg/L.
The preferred MS+6-BA3.0mg/L+NAA0.2mg/L of described adventitious bud proliferation medium.
Described strong seedling culture base comprises MS+6-BA0.2-1.0mg/L+NAA0.1-0.2mg/L.
The preferred MS+6-BA0.2mg/L+NAA0.1mg/L of described strong seedling culture base.
Described root media comprises MS+NAA0.05-0.2mg/L.
The preferred MS+NAA0.1mg/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 floral leaf pampasgrass 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 bud of sprouting spring, remove the blade of coated outside after, 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 axillalry bud inducing culture that comprises MS+6-BA1.0mg/L+NAA0.1mg/L;
(2) differentiation of bud and propagation
Sections was inoculated on the axillalry bud inducing culture after 1 week, and the axillalry bud position begins to expand, and green projection occurs; The visible bud meristematic tissue in 3 week backs was cultivated 1 month again, and little indefinite bud can be grown 3cm; Indefinite bud downcut to change in the adventitious bud proliferation medium that comprises MS+6-BA0.5mg/L+NAA0.1mg/L 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 in medium, to grow;
(3) indefinite bud strong seedling culture
On the adventitious bud proliferation medium, 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-BA0.2mg/L+NAA0.1mg/L and grow, indefinite bud extends rapidly, can grow 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+NAA0.05mg/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) refining seedling and transplanting
Culture of rootage 15 days when root system grows to 0.5cm, is selected the aseptic seedling of well developed root system, robust growth; Refine seedling 2 days 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 bud of sprouting spring, remove the blade of coated outside after, 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 axillalry bud inducing culture that comprises MS+6-BA2.0mg/L+NAA0.2mg/L;
(2) differentiation of bud and propagation
Sections was inoculated on the axillalry bud inducing culture after 2 weeks, and the axillalry bud position begins to expand, and green projection occurs; The visible bud meristematic tissue in 4 week backs was cultivated 1 month again, and little indefinite bud can be grown 4cm; Indefinite bud downcut to change in the adventitious bud proliferation medium that comprises MS+6-BA3.0mg/L+NAA0.2mg/L 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 in medium, to grow;
(3) indefinite bud strong seedling culture
On the adventitious bud proliferation medium, 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-BA0.2mg/L+NAA0.1mg/L and grow, indefinite bud extends rapidly, can grow 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+NAA0.1mg/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) refining seedling and transplanting
Culture of rootage 20 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 bud of sprouting spring, remove the blade of coated outside after, 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 axillalry bud inducing culture that comprises MS+6-BA5.0mg/L+NAA0.5mg/L;
(2) differentiation of bud and propagation
Sections was inoculated on the axillalry bud inducing culture after 3 weeks, and the axillalry bud position begins to expand, and green projection occurs; The visible bud meristematic tissue in 5 week backs was cultivated 2 months again, and little indefinite bud can be grown 4cm; Indefinite bud downcut to change in the adventitious bud proliferation medium that comprises MS+6-BA4.0mg/L+NAA0.2mg/L 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 in medium, to grow;
(3) indefinite bud strong seedling culture
On the adventitious bud proliferation medium; Every clump has 3 strains to extend in the bud of growing thickly that induces; 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-BA1.0mg/L+NAA0.2mg/L and grows; Indefinite bud extends rapidly, can grow 3.5cm after 30 days;
(4) culture of rootage
Get the indefinite bud plantlet of 3.5cm, 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 92%;
(5) refining seedling and transplanting
Culture of rootage 30 days when root system grows to 0.8cm, 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 96%.
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 floral leaf pampasgrass is characterized in that, this method may further comprise the steps:
(1) acquisition of sterilizable material
Get the young tender bud of sprouting spring, remove the blade of coated outside after, 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 axillalry bud inducing culture;
(2) differentiation of bud and propagation
Sections is inoculated in that 1-3 is after week on the axillalry bud inducing culture, and the axillalry bud position begins to expand, and green projection occurs; 3-5 sees the bud meristematic tissue after week, cultivated 1-2 month again, and little indefinite bud is long to 3-4cm; Indefinite bud downcut to change over to carry out enrichment culture in the adventitious bud proliferation medium, the callus of indefinite bud base portion is more, but does not influence propagation; The indefinite bud growth is rapid and do not have the vitrifying bad phenomenon, and it is good in medium, to grow;
(3) indefinite bud strong seedling culture
On the adventitious bud proliferation medium, 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, long 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 5-15 days, grows to 4-6cm 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 15-30 days, when root system grows to 0.5-1cm, select the aseptic seedling of well developed root system, robust growth; Refine seedling 2-4 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 axillalry bud inducing culture comprises MS+6-BA1.0-5.0mg/L+NAA0.1-0.5mg/L;
Described adventitious bud proliferation medium comprises MS+6-BA0.5-4.0mg/L+NAA0.1-0.2mg/L;
Described strong seedling culture base comprises MS+6-BA0.2-1.0mg/L+NAA0.1-0.2mg/L;
Described root media comprises MS+NAA0.05-0.2mg/L.
2. the method for tissue culture of floral leaf pampasgrass according to claim 1 is characterized in that, described axillalry bud inducing culture comprises MS+6-BA2.0mg/L+NAA0.2mg/L.
3. the method for tissue culture of floral leaf pampasgrass according to claim 1 is characterized in that, described adventitious bud proliferation medium comprises MS+6-BA3.0mg/L+NAA0.2mg/L.
4. the method for tissue culture of floral leaf pampasgrass according to claim 1 is characterized in that, described strong seedling culture base comprises MS+6-BA0.2mg/L+NAA0.1mg/L.
5. the method for tissue culture of floral leaf pampasgrass according to claim 1 is characterized in that, described root media comprises MS+NAA0.1mg/L.
6. the method for tissue culture of floral leaf pampasgrass 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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| CN2009100498548A CN101869053B (en) | 2009-04-23 | 2009-04-23 | Tissue culture method of floral leaf pampasgrass |
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| CN110537490A (en) * | 2019-09-20 | 2019-12-06 | 上海上房园林植物研究所有限公司 | method for rapidly breeding floral leaf illicium verum discs through tissue culture |
| CN111084108B (en) * | 2020-02-18 | 2022-05-17 | 美尚生态景观股份有限公司 | Culture method for tissue culture of floral leaf dwarf pampasgrass |
| CN115088619B (en) * | 2022-07-11 | 2023-02-24 | 海南茗卉农林科技发展有限公司 | Tissue culture method for solving stem tip meristem flattening in plant tissue culture |
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Non-Patent Citations (3)
| Title |
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| 刘忠荣, 陈屏昭.植物组织培养技术在观赏植物中的应用.《昭通师范高等专科学校学报》.2003,第25卷(第5期), * |
| 唐道城,梁顺祥.观赏植物组织培养研究进展.《青海大学学报( 自然科学版)》.2006,第24卷(第4期), * |
| 谭小勇，等.蒲苇.《园林》.2007,(第11期), * |
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Effective date of registration: 20140103 Address after: 201114 Pujiang village, Pujiang Town, Shanghai, Minhang District Patentee after: Shanghai Shangfang Landscape Plant Institute 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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Correction item: Patentee Correct: Shanghai Main Garden Plant Research Institute Co., Ltd.| 201114. Pujiang village, Pujiang Town, Shanghai, Minhang District|Shanghai Shanghai Municipal Electric Power Company power supply design Co., Ltd. False: Shanghai Main Garden Plant Research Institute| 201114. Pujiang village, Pujiang Town, Shanghai, Minhang District|Shanghai Shanghai Municipal Electric Power Company power supply design Co., Ltd. Number: 05 Volume: 30 |
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Free format text: CORRECT: PATENTEE; FROM: SHANGHAI SHANGFANG LANDSCAPE PLANT INSTITUTE:201114 MINHANG, SHANGHAI; STATE GRID SHANGHAI ELECTRIC POWER COMPANY, POWER SUPPLY DESIGN (SHANGHAI) CO., LTD. TO: SHANGHAI SHANGFANG LANDSCAPE PLANT RESEARCH INSTITUTE CO., LTD.:201114 MINHANG, SHANGHAI; STATE GRID SHANGHAI ELECTRIC POWER COMPANY, POWER SUPPLY DESIGN (SHANGHAI) CO., LTD. |
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