CN101869071B - Tissue culture method of marsh grass - Google Patents
Tissue culture method of marsh grass Download PDFInfo
- Publication number
- CN101869071B CN101869071B CN2009100499790A CN200910049979A CN101869071B CN 101869071 B CN101869071 B CN 101869071B CN 2009100499790 A CN2009100499790 A CN 2009100499790A CN 200910049979 A CN200910049979 A CN 200910049979A CN 101869071 B CN101869071 B CN 101869071B
- Authority
- CN
- China
- Prior art keywords
- bud
- culture
- root
- days
- seedling
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Landscapes
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention relates to a tissue culture method of marsh grass. The method comprises the steps of acquisition of aseptic materials, differentiation and proliferation of buds, healthy seedling culture of adventitious buds, rooting culture, seedling hardening, transplanting and the like. Compared with the prior art, the method has the advantages of improving the reproductive rate of the marsh grass and the uniformity of the seedlings and realizing industrialized mass production of the 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 marsh grass.
Background technology
Marsh grass is a grass family marsh grass platymiscium, and compactness is grown thickly, Gao Keda 90cm, hat width of cloth 45cm.Leaf adularescent striped, long 17-25cm, scape is compact upright, is with arch slightly, and brown conical inflorescence and green blade are with distinct contrast, and when blooming, the overall appearance of grass is more straight and upright.Sunny or the half cloudy environment of this grass happiness can anti-slight arid or slight wetland, and diseases and insect pests resistance is strong.Autumn, cursive script lower blade meeting yellowing or light red, sight is stronger.Can be used for cultivating in afforestation, bank or the garden, have good annidation and ornamental value.But as the external new varieties of introducing, this plant introduce a fine variety negligible amounts, the seedling supply is restricted.
Summary of the invention
The object of the invention is exactly the method for tissue culture that a kind of marsh grass of the regularity that improves reproduction speed and seedling is provided in order to overcome the defective that above-mentioned prior art exists.
The object of the invention can be realized through following technical scheme:
The method for tissue culture of marsh grass is characterized in that, this method may further comprise the steps:
(1) acquisition of sterilizable material
The budlet that get the marsh grass of sprouting spring, remove blade, wash 1-3h with running water; On superclean bench, utilize the alcohol immersion 10-50s of mass concentration for 70-75% successively again, 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 bottom set surface, bottom set is cut into the sections of the band axillalry bud of 0.5-2cm, 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; 3-4 can see the bud meristematic tissue on the sections after week, cultivates 1-2 month again, and it is long that bud can be grown 3-4cm; With carrying out enrichment culture in the indefinite bud cutting-out immigration adventitious bud proliferation medium, the callus of indefinite bud base portion is more, but indefinite bud is still bred comparatively fast; It is good to grow, and does not have bad phenomenon such as vitrifying;
(3) indefinite bud strong seedling culture
The indefinite bud that in the adventitious bud proliferation medium, induces is in the dwarfing state, and indefinite bud is divided into Xiao Cong, will divide subcaespitose indefinite bud to go on the strong seedling culture base then, and indefinite bud extends rapidly, can grow 3-4cm after 20-30 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 base section of plantlet seedling dissolves the root original hase of white after 8-15 days, can grow to 4-6cm after 25-35 days, and root system is sturdy, and fibrous root is numerous, and rooting rate is 90-100%;
(5) refining seedling and transplanting
At culture of rootage 15-25 days, when root system grows to 0.5-2cm, select the aseptic seedlings of well developed root system, robust growth; Refine seedling 1-3 days in indoor uncork, then seedling is taken out, clean the agar of its root; Plant in the seedbed in the greenhouse and tamed 25-40 days; Can transplant outdoorly, and give rich water quality management, transplanting survival rate is 90-100%.
Described axillalry bud inducing culture comprises MS+6-BA1.0-3.0mg/L+NAA0.05-0.2mg/L.
Described adventitious bud proliferation medium comprises MS+6-BA0.5-2.0mg/L+NAA0.05-0.2mg/L.
The preferred MS+6-BA2.0mg/L+NAA0.2mg/L of described adventitious bud proliferation medium.
Described strong seedling culture base comprises MS+6-BA0.05-2.0mg/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+NAA0.1mg/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 marsh grass and the regularity of seedling through tissue culture technique, 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
The budlet that get the marsh grass of sprouting spring, remove blade, wash 1h with running water; Again on superclean bench, utilizing mass concentration successively is 70% alcohol immersion 10s, and volumetric concentration is that 0.5 ‰ mercury soaks 10min; Use aseptic water washing again 4 times; After utilizing aseptic filter paper to blot the moisture on bottom set surface, bottom set is cut into the sections of the band axillalry bud of 0.5cm, sections is inoculated on the axillalry bud inducing culture that comprises MS+6-BA1.0mg/L+NAA0.05mg/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; Can see the bud meristematic tissue on the sections after 3 weeks, cultivate again 1 month that it is long that bud can be grown 3cm; Indefinite bud downcut moved in the adventitious bud proliferation medium that comprises MS+6-BA0.5mg/L+NAA0.05mg/L carry out enrichment culture, the callus of indefinite bud base portion is more, but indefinite bud is still bred comparatively fast; It is good to grow, and does not have bad phenomenon such as vitrifying;
(3) indefinite bud strong seedling culture
The indefinite bud that in the adventitious bud proliferation medium, induces is in the dwarfing state; Indefinite bud is divided into Xiao Cong; To divide subcaespitose indefinite bud to go on the strong seedling culture base that comprises MS+6-BA0.05mg/L+NAA0.1mg/L then, indefinite bud extends rapidly, can grow 3cm after 20 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 base section of plantlet seedling dissolves the root original hase of white after 8 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, select the aseptic seedlings of well developed root system, robust growth; Refine seedling 1 day in indoor uncork, then seedling is taken out, clean the agar of its root; Plant in the seedbed in the greenhouse and tamed 25 days; Can transplant outdoorly, and give rich water quality management, transplanting survival rate is 90%.
The medium of above-mentioned various situation also comprises sucrose 20g/L, agar powder 4g/L, medium pH 5.5,24 ℃ of cultivation temperature, illumination 1500lx.
Embodiment 2
(1) acquisition of sterilizable material
The budlet that get the marsh grass of sprouting spring, remove blade, wash 2h with running water; Again on superclean bench, utilizing mass concentration successively is 75% alcohol immersion 30s, and volumetric concentration is that 1 ‰ mercury soaks 15min; Use aseptic water washing again 5 times; After utilizing aseptic filter paper to blot the moisture on bottom set surface, bottom set is cut into the sections of the band axillalry bud of 1cm, sections is inoculated on the axillalry bud inducing culture that comprises MS+6-BA2.0mg/L+NAA0.1mg/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; Can see the bud meristematic tissue on the sections after 4 weeks, cultivate 1 first quarter moon again, it is long that bud can be grown 4cm; Indefinite bud downcut moved in the adventitious bud proliferation medium that comprises MS+6-BA2.0mg/L+NAA0.2mg/L carry out enrichment culture, the callus of indefinite bud base portion is more, but indefinite bud is still bred comparatively fast; It is good to grow, and does not have bad phenomenon such as vitrifying;
(3) indefinite bud strong seedling culture
The indefinite bud that in the adventitious bud proliferation medium, induces is in the dwarfing state; Indefinite bud is divided into Xiao Cong; To divide subcaespitose indefinite bud to go on the strong seedling culture base that comprises MS+6-BA1.0mg/L+NAA0.1mg/L then, 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 base section of plantlet seedling 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, select the aseptic seedlings of well developed root system, robust growth; Refine seedling 3 days in indoor uncork, then seedling is taken out, clean the agar of its root; Plant in the seedbed in the greenhouse and tamed 30 days; Can transplant outdoorly, and give rich water quality management, transplanting survival rate is 100%.
The medium of above-mentioned various situation also comprises sucrose 30g/L, agar powder 6g/L, medium pH 5.8,25 ℃ of cultivation temperature, illumination 2000lx.
Embodiment 3
(1) acquisition of sterilizable material
The budlet that get the marsh grass of sprouting spring, remove blade, wash 3h with running water; Again on superclean bench, utilizing mass concentration successively is 75% alcohol immersion 50s, and volumetric concentration is that 2 ‰ mercury soaks 30min; Use aseptic water washing again 6 times; After utilizing aseptic filter paper to blot the moisture on bottom set surface, bottom set is cut into the sections of the band axillalry bud of 2cm, sections is inoculated on the axillalry bud inducing culture that comprises MS+6-BA3.0mg/L+NAA0.2mg/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; Can see the bud meristematic tissue on the sections after 4 weeks, cultivate again 2 months that it is long that bud can be grown 4cm; Indefinite bud downcut moved in the adventitious bud proliferation medium that comprises MS+6-BA2.0mg/L+NAA0.2mg/L carry out enrichment culture, the callus of indefinite bud base portion is more, but indefinite bud is still bred comparatively fast; It is good to grow, and does not have bad phenomenon such as vitrifying;
(3) indefinite bud strong seedling culture
The indefinite bud that in the adventitious bud proliferation medium, induces is in the dwarfing state; Indefinite bud is divided into Xiao Cong; To divide subcaespitose indefinite bud to go on the strong seedling culture base that comprises MS+6-BA2.0mg/L+NAA0.2mg/L then, indefinite bud extends rapidly, can grow 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 base section of plantlet seedling dissolves the root original hase of white after 15 days, can grow to 6cm after 35 days, and root system is sturdy, and fibrous root is numerous, and rooting rate is 95%;
(5) refining seedling and transplanting
Culture of rootage 25 days, when root system grows to 2cm, select the aseptic seedlings of well developed root system, robust growth; Refine seedling 3 days in indoor uncork, then seedling is taken out, clean the agar of its root; Plant in the seedbed in the greenhouse and tamed 40 days; Can transplant outdoorly, and give rich water quality management, transplanting survival rate is 98%.
The medium of above-mentioned various situation also comprises sucrose 40g/L, agar powder 8g/L, medium pH 6.0,26 ℃ of cultivation temperature, illumination 2500lx.
Claims (5)
1. the method for tissue culture of marsh grass is characterized in that, this method may further comprise the steps:
(1) acquisition of sterilizable material
The budlet that get the marsh grass of sprouting spring, remove blade, wash 1-3h with running water; On superclean bench, utilize the alcohol immersion 10-50s of mass concentration for 70-75% successively again, 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 bottom set surface, bottom set is cut into the sections of the band axillalry bud of 0.5-2cm, 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; 3-4 sees the bud meristematic tissue on the sections after week, cultivated 1-2 month again, and bud is long long to 3-4cm; With carrying out enrichment culture in the indefinite bud cutting-out immigration adventitious bud proliferation medium, the callus of indefinite bud base portion is more, but indefinite bud is still bred comparatively fast; It is good to grow, and does not have the vitrifying bad phenomenon;
(3) indefinite bud strong seedling culture
The indefinite bud that in the adventitious bud proliferation medium, induces is in the dwarfing state, and indefinite bud is divided into Xiao Cong, will divide subcaespitose indefinite bud to go on the strong seedling culture base then, and indefinite bud extends rapidly, and is long to 3-4cm after 20-30 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 base section of plantlet seedling dissolves the root original hase of white after 8-15 days, grows to 4-6cm after 25-35 days, and root system is sturdy, and fibrous root is numerous, and rooting rate is 90-100%;
(5) refining seedling and transplanting
At culture of rootage 15-25 days, when root system grows to 0.5-2cm, select the aseptic seedlings of well developed root system, robust growth; Refine seedling 1-3 days in indoor uncork, then seedling is taken out, clean the agar of its root; Plant in the seedbed in the greenhouse and tamed 25-40 days; Promptly transplant outdoorly, and give rich water quality management, transplanting survival rate is 90-100%;
Described axillalry bud inducing culture comprises MS+6-BA1.0-3.0mg/L+NAA0.05-0.2mg/L;
Described adventitious bud proliferation medium comprises MS+6-BA0.5-2.0mg/L+NAA0.05-0.2mg/L;
Described strong seedling culture base comprises MS+6-BA0.05-2.0mg/L+NAA0.1-0.2mg/L;
Described root media comprises MS+NAA0.1-0.3mg/L.
2. the method for tissue culture of marsh grass according to claim 1 is characterized in that, described adventitious bud proliferation medium comprises MS+6-BA2.0mg/L+NAA0.2mg/L.
3. the method for tissue culture of marsh grass according to claim 1 is characterized in that, described strong seedling culture base comprises MS+6-BA1.0mg/L+NAA0.1mg/L.
4. the method for tissue culture of marsh grass according to claim 1 is characterized in that, described root media comprises MS+NAA0.1mg/L.
5. the method for tissue culture of marsh grass 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.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009100499790A CN101869071B (en) | 2009-04-24 | 2009-04-24 | Tissue culture method of marsh grass |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009100499790A CN101869071B (en) | 2009-04-24 | 2009-04-24 | Tissue culture method of marsh grass |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101869071A CN101869071A (en) | 2010-10-27 |
| CN101869071B true CN101869071B (en) | 2012-01-04 |
Family
ID=42994304
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2009100499790A Expired - Fee Related CN101869071B (en) | 2009-04-24 | 2009-04-24 | Tissue culture method of marsh grass |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101869071B (en) |
-
2009
- 2009-04-24 CN CN2009100499790A patent/CN101869071B/en not_active Expired - Fee Related
Non-Patent Citations (3)
| Title |
|---|
| 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),41-43，46. * |
| 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),5-9. * |
| 宋希强，等.浅析观赏草在园林中的运用.《中国园林》.2004,(第3期),32-36. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101869071A (en) | 2010-10-27 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102265785B (en) | Tissue culturing method of hemerocallis middendorfii poinsettia | |
| CN101869062B (en) | Tissue culture method of Hemerocallis dumortieri | |
| CN101869070A (en) | Tissue culture method of pink champagne clematis | |
| CN101869061B (en) | Tissue culture method of ophiopogon planiscapus | |
| CN103734020A (en) | Tissue culture method for German iris tectorum | |
| CN101869054B (en) | Tissue culture method for pennisetum setaceum 'Rubrum' | |
| CN101869066B (en) | Tissue culture method of African agapanthus | |
| CN101869072B (en) | Tissue culture method of golden-heart Iris pseudacorus | |
| CN102273403A (en) | Tissue culture method of nandina domestica firepower | |
| CN101869055B (en) | Tissue culture method of red flower polygonum | |
| CN101869053B (en) | Tissue culture method of floral leaf pampasgrass | |
| CN101869060B (en) | Tissue culture method for heuchera micrantha 'Palace Purple' | |
| CN105340752A (en) | Method for in vitro culture of hemerocallis middendorfii Trautv. et Mey. tissue | |
| CN101869067B (en) | Tissue culture method for anemone | |
| CN101869059B (en) | Tissue culture method of floral leaf myrtle | |
| CN101869068B (en) | Tissue culture method for tulbaghia violacea | |
| CN102293150B (en) | Method for culturing tissues of Buxus sempervives | |
| CN101869071B (en) | Tissue culture method of marsh grass | |
| CN101869063B (en) | Tissue culture method of golden-leaf yucca | |
| CN101869058B (en) | Tissue culture method of short pampas grass | |
| CN101869065B (en) | Tissue culture method for purple prairie clover | |
| CN102293151B (en) | Method for cultivating gold-leaf sedge | |
| CN101869069B (en) | Tissue culturing method of luosifu crocosmia crocosmiflora | |
| CN101869057B (en) | Tissue culture method of floral leaf fatshedera lizei | |
| CN101869056B (en) | Tissue culture method for humid euphorbia ball |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| ASS | Succession or assignment of patent right |
Owner name: SHANGHAI SHANGFANG LANDSCAPE PLANT RESEARCH INSTIT Free format text: FORMER OWNER: SHANGHAI SHANGFANG LANDSCAPE PLANT INSTITUTE Effective date: 20140107 Owner name: STATE GRID SHANGHAI ELECTRIC POWER COMPANY POWER S Effective date: 20140107 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| TR01 | Transfer of patent right |
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 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20120104 Termination date: 20150424 |
|
| EXPY | Termination of patent right or utility model |