CN101869055B - Tissue culture method of red flower polygonum - Google Patents
Tissue culture method of red flower polygonum Download PDFInfo
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- CN101869055B CN101869055B CN2009100498567A CN200910049856A CN101869055B CN 101869055 B CN101869055 B CN 101869055B CN 2009100498567 A CN2009100498567 A CN 2009100498567A CN 200910049856 A CN200910049856 A CN 200910049856A CN 101869055 B CN101869055 B CN 101869055B
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Abstract
The invention relates to a tissue culture method of red flower polygonum, 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 red flower polygonum 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 red flower polygonum.
Background technology
Red flower polygonum is the polygonaceae arsesmart, is perennial plant, and leaf, stem are spent also to take on a red color slightly redly, is good sight leaf, sight floral material.Have good annidation and ornamental value, can in presbyopic glasses and garden, use.But as the external new varieties of introducing, at the initial stage of introducing a fine variety, red flower polygonum introduce a fine variety negligible amounts, its cuttage, plant division are slow, so 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 red flower polygonum of property stability.
The object of the invention can be realized through following technical scheme:
The method for tissue culture of red flower polygonum is characterized in that, this method may further comprise the steps:
(1) acquisition of sterilizable material
Get the young tender branch of sprouting spring, remove branches and leaves 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 bud of 0.5-2cm again, sections is inoculated on the bud inducing culture;
(2) differentiation of bud and propagation
Sections is inoculated in that 1-3 is after week on the bud inducing culture, and the axillalry bud position begins to expand, and green projection occurs; 2-4 is visible bud meristematic tissue after week, cultivates 1-2 month again, and little indefinite bud can be grown 2-3cm; 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 90-95%.
Described axillalry bud inducing culture comprises MS+6-BA1.0-3.0mg/L+NAA0.1-0.3mg/L.
The preferred MS+6-BA3.0mg/L+NAA0.3mg/L of described axillalry bud inducing culture.
Described adventitious bud proliferation medium comprises MS+6-BA1.0-3.0mg/L+NAA0.1-0.3mg/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-BA0.5-2.0mg/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+NAA0.1mg/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 red flower polygonum 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 branch of sprouting spring, remove branches and leaves 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 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 1 week, and the axillalry bud position begins to expand, and green projection occurs; The visible bud meristematic tissue in 2 week backs was cultivated 1 month again, and little indefinite bud can be grown 2cm; Indefinite bud downcut to change in the adventitious bud proliferation medium that comprises MS+6-BA1.0mg/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-BA0.5mg/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+NAA0.1mg/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 branch of sprouting spring, remove branches and leaves 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 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 2 weeks, 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-BA2.0mg/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-BA1.0mg/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 95%.
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 branch of sprouting spring, remove branches and leaves 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 bud of 2cm 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, and the axillalry bud position begins to expand, and green projection occurs; The visible bud meristematic tissue in 4 week backs was cultivated 2 months again, and little indefinite bud can be grown 3cm; Indefinite bud downcut to change in the adventitious bud proliferation medium that comprises MS+6-BA3.0mg/L+NAA0.3mg/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-BA2.0mg/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+NAA0.3mg/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 95%;
(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 92%.
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 red flower polygonum is characterized in that, this method may further comprise the steps:
(1) acquisition of sterilizable material
Get the young tender branch of sprouting spring, remove branches and leaves 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 bud of 0.5-2cm again, sections is inoculated on the bud inducing culture;
(2) differentiation of bud and propagation
Sections is inoculated in that 1-3 is after week on the bud inducing culture, and the axillalry bud position begins to expand, and green projection occurs; 2-4 sees the bud meristematic tissue after week, cultivated 1-month again, and little indefinite bud is long to 2-3cm; 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 90-95%;
Described bud inducing culture comprises MS+6-BA1.0-3.0mg/L+NAA0.1-0.3mg/L;
Described adventitious bud proliferation medium 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 red flower polygonum according to claim 1 is characterized in that, described bud inducing culture comprises MS+6-BA3.0mg/L+NAA0.3mg/L.
3. the method for tissue culture of red flower polygonum according to claim 1 is characterized in that, described adventitious bud proliferation medium comprises MS+6-BA2.0mg/L+NAA0.2mg/L.
4. the method for tissue culture of red flower polygonum 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 red flower polygonum according to claim 1 is characterized in that, described root media comprises MS+NAA0.1mg/L.
6. the method for tissue culture of red flower polygonum 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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| CN2009100498567A CN101869055B (en) | 2009-04-23 | 2009-04-23 | Tissue culture method of red flower polygonum |
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| CN104686357A (en) * | 2015-03-12 | 2015-06-10 | 朱炳贵 | Establishment method of rapid polygonum capitatum in-vitro propagation system |
| CN109220812A (en) * | 2018-11-25 | 2019-01-18 | 林登淞 | A kind of tissue culture technique of Chinese knotweed |
| CN116058281B (en) * | 2022-12-02 | 2023-11-21 | 甘肃农业大学 | Method for rapid propagation of polygonum mongolicum tissue |
| CN116439131A (en) * | 2023-04-03 | 2023-07-18 | 内蒙古库布其沙漠技术研究院 | Tissue culture seedling raising method for sand-fixing plant polygonum sapidum |
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Non-Patent Citations (5)
| 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. * |
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| 地里拜尔•对山拜,等.观赏植物组织培养技术的探讨.《新疆农业科学》.2002,第39卷(第2期),106-108. * |
| 对山拜,等.观赏植物组织培养技术的探讨.《新疆农业科学》.2002,第39卷(第2期),106-108. |
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Owner name: SHANGHAI SHANGFANG LANDSCAPE PLANT RESEARCH INSTIT Free format text: FORMER OWNER: SHANGHAI SHANGFANG LANDSCAPE PLANT INSTITUTE Effective date: 20140102 Owner name: STATE GRID SHANGHAI ELECTRIC POWER COMPANY POWER S Effective date: 20140102 |
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