CN101869060B - Tissue culture method for heuchera micrantha 'Palace Purple' - Google Patents
Tissue culture method for heuchera micrantha 'Palace Purple' Download PDFInfo
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- CN101869060B CN101869060B CN2009100499682A CN200910049968A CN101869060B CN 101869060 B CN101869060 B CN 101869060B CN 2009100499682 A CN2009100499682 A CN 2009100499682A CN 200910049968 A CN200910049968 A CN 200910049968A CN 101869060 B CN101869060 B CN 101869060B
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Abstract
The invention relates to a tissue culture method for heuchera micrantha 'Palace Purple'. 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 heuchera micrantha 'Palace Purple' and the trimming of the seedlings, and improves the stability of shape and properties of the heuchera micrantha 'Palace Purple', so that the 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 heuchera micrantha ' Palace Purple '.
Background technology
Heuchera micrantha ' Palace Purple ' is the Saxifragaceae heuchera.Originate in the North America, be perennial perennial root flowers, leaf is aubergine, and happiness half is shady, anti-full light; Be used for sylvan life flower border in the gardens more, the ground quilt, courtyard greening or the like, the florescence is the 4-10 month, is desirable look leaf perennial root flower border material.But as the external new varieties of introducing, this plant introduction negligible amounts, division propagation is slow, and the demand on the market receives certain restriction because of the seedling supply.
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 keeps the method for tissue culture of the heuchera micrantha ' Palace Purple ' of original maternal character.
The object of the invention can be realized through following technical scheme:
The method for tissue culture of heuchera micrantha ' Palace Purple ' is characterized in that, this method may further comprise the steps:
(1) acquisition of sterilizable material
Get the young tender bud of sprouting, 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 the surperficial moisture of bud after; Bud is cut into the sections of the long band axillalry bud of 0.5-2cm, 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; 3-5 is visible bud meristematic tissue after week, cultivates budlet Cheng Cong again 1-2 month; Indefinite bud downcut to change over to carry out enrichment culture in the adventitious bud proliferation medium,, do not influence the propagation of indefinite bud though the base portion of the bud of differentiation has more callus; 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 3-5 strain to extend in the bud of growing thickly that induces, and remaining bud of growing thickly is in the dwarfing state; After the bud of will growing thickly is divided into Xiao Cong; Go on the strong seedling culture base and grow, the indefinite bud elongation can be grown to 1-2cm after 15-30 days rapidly;
(4) culture of rootage
Get the indefinite bud plantlet of 1-2cm, 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 1-2cm 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
Culture of rootage 30-35 days, when root system grows to 2-3cm, select the aseptic seedling of well developed root system, robust growth; Refine seedling 1-3 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 bud inducing culture comprises MS+6-BA1.0-3.0mg/L+NAA0.1-0.3mg/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-BA0.2-2.0mg/L+NAA0.05-0.2mg/L.
The preferred MS+6-BA0.2mg/L+NAA0.05mg/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 heuchera micrantha ' Palace Purple ' and the regularity of seedling, and has kept original maternal character better 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, 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 bud surface after; Bud is cut into the sections of the long band axillalry bud of 0.5cm, 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 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 budlet Cheng Cong again 1 month; Indefinite bud downcut to change in the adventitious bud proliferation medium that comprises MS+6-BA0.5mg/L+NAA0.05mg/L carry out enrichment culture,, do not influence the propagation of indefinite bud though the base portion of the bud of differentiation has more callus; 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 bud of growing thickly 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-BA0.2mg/L+NAA0.05mg/L and grows; The indefinite bud elongation can be grown to 1cm after 15 days rapidly;
(4) culture of rootage
Get the indefinite bud plantlet of 1cm, 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 1cm 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 30 days when root system grows to 2cm, 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 2g/L, pH=5.5,22 ℃ of cultivation temperature, illumination 1500lx.
Embodiment 2
(1) acquisition of sterilizable material
Get the young tender bud of sprouting, 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 bud surface after; Bud is cut into the sections of the long band axillalry bud of 1cm, is inoculated on the 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 3 week backs was cultivated budlet Cheng Cong again 1 month; 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,, do not influence the propagation of indefinite bud though the base portion of the bud of differentiation has more callus; 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 5 strains to extend in the bud of growing thickly that induces; Remaining bud of growing thickly 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-BA0.2mg/L+NAA0.05mg/L and grows; The indefinite bud elongation can be grown to 1.5cm after 20 days rapidly;
(4) culture of rootage
Get the indefinite bud plantlet of 1.5cm, 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 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 3cm, 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 bud of sprouting, 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 bud surface after; Bud is cut into the sections of the long band axillalry bud of 2cm, 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 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 budlet Cheng Cong again 2 months; 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,, do not influence the propagation of indefinite bud though the base portion of the bud of differentiation has more callus; 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 4 strains to extend in the bud of growing thickly that induces; Remaining bud of growing thickly 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; The indefinite bud elongation can be grown to 2cm after 30 days rapidly;
(4) culture of rootage
Get the indefinite bud plantlet of 2cm, 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 2cm after 35 days, and root system is sturdy, and fibrous root is numerous, and rooting rate is 96%;
(5) refining seedling and transplanting
Culture of rootage 35 days when root system grows to 3cm, 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 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 (5)
1. the method for tissue culture of heuchera micrantha ' Palace Purple ' is characterized in that, this method may further comprise the steps:
(1) acquisition of sterilizable material
Get the young tender bud of sprouting, 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 the surperficial moisture of bud after; Bud is cut into the sections of the long band axillalry bud of 0.5-2cm, 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; 3-5 sees the bud meristematic tissue after week, cultivated budlet Cheng Cong again 1-2 month; Indefinite bud downcut to change over to carry out enrichment culture in the adventitious bud proliferation medium,, do not influence the propagation of indefinite bud though the base portion of the bud of differentiation has more callus; 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 3-5 strain to extend in the bud of growing thickly that induces, and remaining bud of growing thickly 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 the indefinite bud elongation is grown to 1-2cm after 15-30 days rapidly;
(4) culture of rootage
Get the indefinite bud plantlet of 1-2cm, 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 1-2cm 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
Culture of rootage 30-35 days, when root system grows to 2-3cm, select the aseptic seedling of well developed root system, robust growth; Refine seedling 1-3 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-BA0.5-2.0mg/L+NAA0.05-0.2mg/L;
Described strong seedling culture base comprises MS+6-BA0.2-2.0mg/L+NAA0.05-0.2mg/L;
Described root media comprises MS+NAA0.1-0.3mg/L.
2. the method for tissue culture of heuchera micrantha ' Palace Purple ' 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 heuchera micrantha ' Palace Purple ' according to claim 1 is characterized in that, described strong seedling culture base comprises MS+6-BA0.2mg/L+NAA0.05mg/L.
4. the method for tissue culture of heuchera micrantha ' Palace Purple ' according to claim 1 is characterized in that, described root media comprises MS+NAA0.1mg/L.
5. the method for tissue culture of heuchera micrantha ' Palace Purple ' 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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| CN2009100499682A CN101869060B (en) | 2009-04-24 | 2009-04-24 | Tissue culture method for heuchera micrantha 'Palace Purple' |
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| CN106942051B (en) * | 2017-03-02 | 2019-03-01 | 浙江省萧山棉麻研究所 | A kind of culture medium and propagation method of the tissue-culturing quick-propagation of alum root blade |
| CN108513911A (en) * | 2018-07-02 | 2018-09-11 | 杭州市园林绿化股份有限公司 | A kind of regenerated tissue cultures abductive approach of alum root petiole adventitious bud high frequency |
| CN111513061B (en) * | 2020-05-22 | 2022-05-03 | 上海市农业生物基因中心 | Ultralow-temperature preservation and recovery culture method for alum root clump buds |
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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. * |
| 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. * |
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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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