CN102273405B - Cultivating method of Hosta plantaginea Gold Standard - Google Patents
Cultivating method of Hosta plantaginea Gold Standard Download PDFInfo
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Abstract
The invention relates to a cultivating method of Hosta plantaginea Gold Standard. The method comprises the following steps of: first carrying out sterile treatment on Hosta plantaginea Gold Standard buds excavated in the early spring; and then inoculating the buds on a bud inducing culture medium to differentiate the buds; culturing the differentiated buds for 1 month; and then cutting off a budding callus and putting the budding callus into a proliferation medium to be proliferated so as to obtain cluster buds; after dividing the cluster buds into shoots or single buds, putting the shoots or the single buds into a seedling culture medium to culture the seedling with adventitious buds; obtaining 2-3 cm small plants after 20 days; transferring the small plants into a rooting culture medium to be induced for rooting; selecting strongly growing sterile seedlings with developed roots; carrying out seedling adaptation indoors in an open bottle for three days; washing off agars at the roots of the seedlings after taking out the seedlings; transplanting the seedlings outdoors after naturalizing the seedlings in a greenhouse for 40 days; and carrying out fertilizer and water management on the seedlings. Compared with the prior art, the cultivating method of the Hosta plantaginea Gold Standard provided by the invention has the following advantages that: the reproductive speed and the evenness of the nursery-grown plants can be largely increased; the original maternal character is preferably kept; and the survival rate can be increased so that the yield is increased and the transplanting survival rate can reach 95%.
Description
Technical field
The present invention relates to a kind of herbal method for tissue culture, especially relate to the breeding method of a kind of jade hairpin Jin Qi.
Background technology
Jade hairpin perennial root draft.Plant height 30cm-50cm.Phyllopodium generates clump, and is avette avette to heart shape; Give birth on the raceme top, is higher than leafage, and flower is white, tubulose infundibulate, giving off a strong fragrance.June at florescence-August.Jade hairpin ' Jin Qi ' is the garden-variety that jade hairpin obtains through artificially breeding, the leaf yellow green, and the edge is blackish green, presents elegant leaf line, and sight is good; Property is strong, cold-resistant, happiness is cloudy, avoids the sunlight direct projection, do not select soil, but with good, the fertile moistening place of draining grow prolifically, be shade plant preferably, in gardens, can be used for doing ground cover plant under the tree, or plant in rock garden or building north side, also can potted plantly view and admire or do cut-flower usefulness, can do the sparse woods ground cover plant again, be good presbyopic glasses and garden material.Because of the kind as external introduction, sight is special, negligible amounts, and plant division speed is slow, but the huge market demand, so the seedling supply is restricted.By tissue culture technology, greatly improve the regularity of reproduction speed and seedling, keep original maternal character better.Being reported in of jade hairpin ' Jin Qi ' group training domesticly do not seen as yet.
Summary of the invention
Purpose of the present invention is exactly to provide a kind of regularity that can greatly improve reproduction speed and seedling for the defective that overcomes above-mentioned prior art existence, keeps the breeding method of a kind of jade hairpin Jin Qi of original maternal character better.
Purpose of the present invention can be achieved through the following technical solutions:
The breeding method of a kind of jade hairpin Jin Qi, it is characterized in that, this method is that jade hairpin " Jin Qi " budlet that will take the early spring at first carries out the asepticize processing, be inoculated in the differentiation of carrying out bud on the budlet inducing culture then, cultivated 1 month, downcutting band bud callus then puts into proliferated culture medium and breeds, obtain the bud of growing thickly, after the bud of will growing thickly is divided into Xiao Cong or simple bud, puts into the strong seedling culture base and carry out the indefinite bud strong seedling culture, grow up to the plantlet of 2-3cm after 20 days, with the root induction in the root media of transferring of this plantlet, when root system grows to the 1-2cm left and right sides, select the aseptic seedling of well developed root system robust growth, indoor uncork hardening 3 days, take out afterwash root agar, domestication can be transplanted outdoorly after 40 days in the greenhouse, gave rich water quality management and got final product.
It is that jade hairpin " Jin Qi " budlet that will take the early spring is placed on the superclean bench with running water flushing 2h that described asepticize is handled, with concentration is the alcohol immersion 20-40s of 75wt%, the mercuric chloride solution of 1wt ‰ soaks 10-20min, blots surperficial moisture content with aseptic filter paper behind aseptic water washing 5-6 time.
The differentiation of described bud is the budlet base portion after the asepticize processing to be cut into 1cm length be inoculated on the budlet inducing culture, and after 3 weeks, the budlet base portion begins to expand, and the yellow green projection occurs, the visible tangible budlet callus in 2 week backs.
The composition of described budlet inducing culture is MS+6-BA1.0mg/L+NAA0.1mg/L, MS+6-BA3.0mg/L+NAA0.3mg/L or MS+6-BA5.0mg/L+NAA0.5mg/L, preferred MS+6-BA5.0mg/L+NAA0.5mg/L.
The composition of described proliferated culture medium is MS+6-BA1.0mg/L+NAA0.1mg/L, MS+6-BA2.0mg/L+NAA0.2mg/L or MS+6-BA3.0mg/L+NAA0.3mg/L, preferred MS+6-BA2.0mg/L+NAA0.2mg/L.
The composition of described strong seedling culture base is MS+6-BA0.5mg/L+NAA0.1mg/L, MS+6-BA1.0mg/L+NAA0.1mg/L or MS+6-BA2.0mg/L+NAA0.2mg/L.
The preferred MS+6-BA1.0mg/L+NAA0.1mg/L of the composition of described strong seedling culture base.
The composition of described root media is MS+NAA0.05mg/L, MS+NAA0.1mg/L or MS+NAA0.2mg/L.
The preferred MS+NAA0.1mg/L of the composition of described root media.
Described budlet inducing culture, proliferated culture medium, strong seedling culture base and culture of rootage based component also comprise sucrose 30g/L, agar 6g/L, and the cultivation temperature of this medium is 24-26 ℃, and illumination is 70-90 μ mol/ms, and the pH value is 5.5-6.0.
Compared with prior art, the present invention can improve the regularity of reproduction speed and nursery stock greatly by tissue culture technology, keeps original maternal character better, and can improve survival rate, thereby improve output.
Embodiment
The present invention is described in detail below in conjunction with specific embodiment.
Embodiment 1
The breeding method of a kind of jade hairpin Jin Qi, this method may further comprise the steps:
(1) asepticize is handled
The jade hairpin that takes the early spring " Jin Qi " budlet, with behind the running water flushing 2h on superclean bench, with concentration is the alcohol immersion 30s of 75wt%, the mercuric chloride solution of 1wt ‰ soaks 15min, behind aseptic water washing 5-6 time, blot surperficial moisture content, get the budlet base portion and be cut into that 1cm is long to be inoculated on the budlet inducing culture MS+6-BA5.0mg/L+NAA0.5mg/L with aseptic filter paper;
(2) differentiation of bud and propagation
Budlet was inoculated on the budlet inducing culture after 3 weeks, the budlet base portion begins to expand, the yellow green projection appears, the visible significantly callus in 2 week backs was cultivated after 1 month, and cutting-out band bud callus is put into proliferated culture medium MS+6-BA2.0mg/L+NAA0.2mg/L and cultivated, the base portion callus is many, but do not influence the propagation of indefinite bud, the indefinite bud growth there is no bad phenomenon such as vitrifying rapidly, and it is good to grow in this cultivation;
(3) indefinite bud strong seedling culture
Induce the bud of growing thickly on proliferated culture medium, every clump of the bud of growing thickly has only the 2-3 strain to extend, and all the other are in the dwarfing state, after the Da Cong bud is divided into Xiao Cong or simple bud, put on the strong seedling culture base MS+6-BA1.0mg/L+NAA0.1mg/L, indefinite bud extends rapidly, can grow 2-3cm after 20 days;
(4) culture of rootage
Get and highly be the plantlet of 2-3cm, root induction among the root media MS+NAA0.1mg/L is gone in switching, and the seedling base section dissolves the root original hase of many whites after 10 days, can grow to 4-6cm after 30 days, and root system is sturdy, and fibrous root is numerous, and rooting rate is 100%;
(5) hardening and transplanting
Culture of rootage 25 days when root system grows to 1-2cm, is selected the aseptic seedling of well developed root system robust growth, indoor uncork hardening 3 days is taken out afterwash root agar, and domestication is after 40 days in the greenhouse, can transplant outdoorly, give rich water quality management and get final product, transplanting survival rate is 95%.
Embodiment 2
The breeding method of a kind of jade hairpin Jin Qi, this method may further comprise the steps:
(1) asepticize is handled
The jade hairpin that takes the early spring " Jin Qi " budlet, with behind the running water flushing 2h on superclean bench, with concentration is the alcohol immersion 20s of 75wt%, the mercuric chloride solution of 1wt ‰ soaks 10min, behind aseptic water washing 5-6 time, blot surperficial moisture content, get the budlet base portion and be cut into that 1cm is long to be inoculated on the budlet inducing culture MS+6-BA1.0mg/L+NAA0.1mg/L with aseptic filter paper;
(2) differentiation of bud and propagation
Budlet was inoculated on the budlet inducing culture after 3 weeks, the budlet base portion begins to expand, the yellow green projection appears, the visible significantly callus in 2 week backs was cultivated after 1 month, and cutting-out band bud callus is put into proliferated culture medium MS+6-BA1.0mg/L+NAA0.1mg/L and cultivated, the base portion callus is many, but do not influence the propagation of indefinite bud, the indefinite bud growth there is no bad phenomenon such as vitrifying rapidly, and it is good to grow in this cultivation;
(3) indefinite bud strong seedling culture
Induce the bud of growing thickly on proliferated culture medium, every clump of the bud of growing thickly has only the 2-3 strain to extend, and all the other are in the dwarfing state, after the Da Cong bud is divided into Xiao Cong or simple bud, put on the strong seedling culture base MS+6-BA0.5mg/L+NAA0.1mg/L, indefinite bud extends rapidly, can grow 2-3cm after 20 days;
(4) culture of rootage
Get and highly be the plantlet of 2-3cm, root induction among the root media MS+NAA0.05mg/L is gone in switching, and the seedling base section dissolves the root original hase of many whites after 10 days, can grow to 2-3cm after 30 days;
(5) hardening and transplanting
Continue culture of rootage 20 days, and when root system grows to 1-2cm, selected the aseptic seedling of well developed root system robust growth, indoor uncork hardening 3 days is taken out afterwash root agar, and domestication is after 40 days in the greenhouse, can transplant outdoorly, give rich water quality management and get final product, transplanting survival rate is 95%.
Embodiment 3
The breeding method of a kind of jade hairpin Jin Qi, this method may further comprise the steps:
(1) asepticize is handled
The jade hairpin that takes the early spring " Jin Qi " budlet, with behind the running water flushing 2h on superclean bench, with concentration is the alcohol immersion 40s of 75wt%, the mercuric chloride solution of 1wt ‰ soaks 20min, behind aseptic water washing 5-6 time, blot surperficial moisture content, get the budlet base portion and be cut into that 1cm is long to be inoculated on the budlet inducing culture MS+6-BA3.0mg/L+NAA0.3mg/L with aseptic filter paper;
(2) differentiation of bud and propagation
Budlet was inoculated on the budlet inducing culture after 3 weeks, the budlet base portion begins to expand, the yellow green projection appears, the visible significantly callus in 2 week backs was cultivated after 1 month, and cutting-out band bud callus is put into proliferated culture medium MS+6-BA3.0mg/L+NAA0.3mg/L and cultivated, the base portion callus is many, but do not influence the propagation of indefinite bud, the indefinite bud growth there is no bad phenomenon such as vitrifying rapidly, and it is good to grow in this cultivation;
(3) indefinite bud strong seedling culture
Induce the bud of growing thickly on proliferated culture medium, every clump of the bud of growing thickly has only the 2-3 strain to extend, and all the other are in the dwarfing state, after the Da Cong bud is divided into Xiao Cong or simple bud, put on the strong seedling culture base MS+6-BA2.0mg/L+NAA0.2mg/L, indefinite bud extends rapidly, can grow 2-3cm after 20 days;
(4) culture of rootage
Get and highly be the plantlet of 2-3cm, root induction among the root media MS+NAA0.2mg/L is gone in switching, and the seedling base section dissolves the root original hase of many whites after 10 days, can grow to 2-3cm after 30 days;
(5) hardening and transplanting
Continue culture of rootage 30 days, and when root system grows to 1-2cm, selected the aseptic seedling of well developed root system robust growth, indoor uncork hardening 3 days is taken out afterwash root agar, and domestication is after 40 days in the greenhouse, can transplant outdoorly, give rich water quality management and get final product, transplanting survival rate is 95%.
Embodiment 4
The breeding method of a kind of jade hairpin Jin Qi, this method may further comprise the steps:
(1) asepticize is handled
The jade hairpin that takes the early spring " Jin Qi " budlet, with behind the running water flushing 2h on superclean bench, with concentration is the alcohol immersion 30s of 75wt%, the mercuric chloride solution of 1wt ‰ soaks 15min, behind aseptic water washing 5-6 time, blot surperficial moisture content, get the budlet base portion and be cut into that 1cm is long to be inoculated on the budlet inducing culture MS+6-BA3.0mg/L+NAA0.3mg/L with aseptic filter paper;
(2) differentiation of bud and propagation
Budlet was inoculated on the budlet inducing culture after 3 weeks, the budlet base portion begins to expand, the yellow green projection appears, the visible significantly callus in 2 week backs was cultivated after 1 month, and cutting-out band bud callus is put into proliferated culture medium MS+6-BA1.0mg/L+NAA0.1mg/L and cultivated, the base portion callus is many, but do not influence the propagation of indefinite bud, the indefinite bud growth there is no bad phenomenon such as vitrifying rapidly, and it is good to grow in this cultivation;
(3) indefinite bud strong seedling culture
Induce the bud of growing thickly on proliferated culture medium, every clump of the bud of growing thickly has only the 2-3 strain to extend, and all the other are in the dwarfing state, after the Da Cong bud is divided into Xiao Cong or simple bud, put on the strong seedling culture base MS+6-BA0.5mg/L+NAA0.1mg/L, indefinite bud extends rapidly, can grow 2-3cm after 20 days;
(4) culture of rootage
Get and highly be the plantlet of 2-3cm, root induction among the root media MS+NAA0.1mg/L is gone in switching, and the seedling base section dissolves the root original hase of many whites after 10 days, can grow to 2-3cm after 30 days, and root system is sturdy, and fibrous root is numerous, and rooting rate is 100%;
(5) hardening and transplanting
Continue culture of rootage 25 days, and when root system grows to 1-2cm, selected the aseptic seedling of well developed root system robust growth, indoor uncork hardening 3 days is taken out afterwash root agar, and domestication is after 40 days in the greenhouse, can transplant outdoorly, give rich water quality management and get final product, transplanting survival rate is 95%.
Described budlet inducing culture, proliferated culture medium, strong seedling culture base and culture of rootage based component also comprise sucrose 30g/L, agar 6g/L, and the cultivation temperature of this medium is 24 ℃, and illumination is 70 μ mol/ms, and the pH value is 5.5.
Embodiment 5
The breeding method of a kind of jade hairpin Jin Qi, this method may further comprise the steps:
(1) asepticize is handled
The jade hairpin that takes the early spring " Jin Qi " budlet, with behind the running water flushing 2h on superclean bench, with concentration is the alcohol immersion 20s of 75wt%, the mercuric chloride solution of 1wt ‰ soaks 10min, behind aseptic water washing 5-6 time, blot surperficial moisture content, get the budlet base portion and be cut into that 1cm is long to be inoculated on the budlet inducing culture MS+6-BA1.0mg/L+NAA0.1mg/L with aseptic filter paper;
(2) differentiation of bud and propagation
Budlet was inoculated on the budlet inducing culture after 3 weeks, the budlet base portion begins to expand, the yellow green projection appears, the visible significantly callus in 2 week backs was cultivated after 1 month, and cutting-out band bud callus is put into proliferated culture medium MS+6-BA2.0mg/L+NAA0.2mg/L and cultivated, the base portion callus is many, but do not influence the propagation of indefinite bud, the indefinite bud growth there is no bad phenomenon such as vitrifying rapidly, and it is good to grow in this cultivation;
(3) indefinite bud strong seedling culture
Induce the bud of growing thickly on proliferated culture medium, every clump of the bud of growing thickly has only the 2-3 strain to extend, and all the other are in the dwarfing state, after the Da Cong bud is divided into Xiao Cong or simple bud, put on the strong seedling culture base MS+6-BA1.0mg/L+NAA0.1mg/L, indefinite bud extends rapidly, can grow 2-3cm after 20 days;
(4) culture of rootage
Get and highly be the plantlet of 2-3cm, root induction among the root media MS+NAA0.05mg/L is gone in switching, and the seedling base section dissolves the root original hase of many whites after 10 days, can grow to 2-3cm after 30 days;
(5) hardening and transplanting
Continue culture of rootage 20 days, and when root system grows to 1-2cm, selected the aseptic seedling of well developed root system robust growth, indoor uncork hardening 3 days is taken out afterwash root agar, and domestication is after 40 days in the greenhouse, can transplant outdoorly, give rich water quality management and get final product, transplanting survival rate is 95%.
Described budlet inducing culture, proliferated culture medium, strong seedling culture base and culture of rootage based component also comprise sucrose 30g/L, agar 6g/L, and the cultivation temperature of this medium is 25 ℃, and illumination is 80 μ mol/ms, and the pH value is 5.8.
Embodiment 6
The breeding method of a kind of jade hairpin Jin Qi, this method may further comprise the steps:
(1) asepticize is handled
The jade hairpin that takes the early spring " Jin Qi " budlet, with behind the running water flushing 2h on superclean bench, with concentration is the alcohol immersion 40s of 75wt%, the mercuric chloride solution of 1wt ‰ soaks 20min, behind aseptic water washing 5-6 time, blot surperficial moisture content, get the budlet base portion and be cut into that 1cm is long to be inoculated on the budlet inducing culture MS+6-BA5.0mg/L+NAA0.5mg/L with aseptic filter paper;
(2) differentiation of bud and propagation
Budlet was inoculated on the budlet inducing culture after 3 weeks, the budlet base portion begins to expand, the yellow green projection appears, the visible significantly callus in 2 week backs was cultivated after 1 month, and cutting-out band bud callus is put into proliferated culture medium MS+6-BA3.0mg/L+NAA0.3mg/L and cultivated, the base portion callus is many, but do not influence the propagation of indefinite bud, the indefinite bud growth there is no bad phenomenon such as vitrifying rapidly, and it is good to grow in this cultivation;
(3) indefinite bud strong seedling culture
Induce the bud of growing thickly on proliferated culture medium, every clump of the bud of growing thickly has only the 2-3 strain to extend, and all the other are in the dwarfing state, after the Da Cong bud is divided into Xiao Cong or simple bud, put on the strong seedling culture base MS+6-BA2.0mg/L+NAA0.2mg/L, indefinite bud extends rapidly, can grow 2-3cm after 20 days;
(4) culture of rootage
Get and highly be the plantlet of 2-3cm, root induction among the root media MS+NAA0.2mg/L is gone in switching, and the seedling base section dissolves the root original hase of many whites after 10 days, can grow to 2-3cm after 30 days;
(5) hardening and transplanting
Continue culture of rootage 30 days, and when root system grows to 1-2cm, selected the aseptic seedling of well developed root system robust growth, indoor uncork hardening 3 days is taken out afterwash root agar, and domestication is after 40 days in the greenhouse, can transplant outdoorly, give rich water quality management and get final product, transplanting survival rate is 95%.
Described budlet inducing culture, proliferated culture medium, strong seedling culture base and culture of rootage based component also comprise sucrose 30g/L, agar 6g/L, and the cultivation temperature of this medium is 26 ℃, and illumination is 90 μ mol/ms, and the pH value is 6.0.
Claims (6)
1. the breeding method of a jade hairpin Jin Qi, it is characterized in that, this method is that jade hairpin " Jin Qi " budlet that will take the early spring at first carries out the asepticize processing, be inoculated in the differentiation of carrying out bud on the budlet inducing culture then, cultivated 1 month, downcutting band bud callus then puts into proliferated culture medium and breeds, obtain the bud of growing thickly, after the bud of will growing thickly is divided into Xiao Cong or simple bud, puts into the strong seedling culture base and carry out the indefinite bud strong seedling culture, grow up to the plantlet of 2-3cm after 20 days, with the root induction in the root media of transferring of this plantlet, when root system grows to the 1-2cm left and right sides, select the aseptic seedling of well developed root system robust growth, indoor uncork hardening 3 days, take out afterwash root agar, domestication can be transplanted outdoorly after 40 days in the greenhouse, gave rich water quality management and got final product;
The differentiation of described bud is the budlet base portion after the asepticize processing to be cut into 1cm length be inoculated on the budlet inducing culture, and after 3 weeks, the budlet base portion begins to expand, and the yellow green projection occurs, the visible tangible budlet callus in 2 week backs;
The composition of described budlet inducing culture is MS+6-BA1.0mg/L+NAA0.1mg/L, MS+6-BA3.0 mg/L+NAA0.3mg/L or MS+6-BA5.0mg/L+NAA0.5mg/L;
The composition of described proliferated culture medium is MS+6-BA1.0mg/L+NAA0.1mg/L, MS+6-BA2.0mg/L+NAA0.2mg/L or MS+6-BA3.0mg/L+NAA0.3mg/L;
The composition of described strong seedling culture base is MS+6-BA0.5mg/L+NAA0.1mg/L, MS+6-BA1.0 mg/L+NAA0.1mg/L or MS+6-BA2.0mg/L+NAA0.2mg/L;
The composition of described root media is MS+NAA0.05mg/L, MS+NAA0.1mg/L or MS+NAA0.2mg/L.
2. the breeding method of a kind of jade hairpin Jin Qi according to claim 1, it is characterized in that, it is that jade hairpin " Jin Qi " budlet that will take the early spring is placed on the superclean bench with running water flushing 2h that described asepticize is handled, with concentration is the alcohol immersion 20-40s of 75wt%, the mercuric chloride solution of 1wt ‰ soaks 10-20min, blots surperficial moisture content with aseptic filter paper behind aseptic water washing 5-6 time.
3. the breeding method of a kind of jade hairpin Jin Qi according to claim 1 is characterized in that, the composition of described budlet inducing culture is MS+6-BA5.0mg/L+NAA0.5mg/L.
4. the breeding method of a kind of jade hairpin Jin Qi according to claim 1 is characterized in that, the composition of described proliferated culture medium is MS+6-BA2.0mg/L+NAA0.2mg/L.
5. the breeding method of a kind of jade hairpin Jin Qi according to claim 1 is characterized in that, the preferred MS+6-BA1.0mg/L+NAA0.1mg/L of the composition of described strong seedling culture base.
6. the breeding method of a kind of jade hairpin Jin Qi according to claim 1 is characterized in that, the preferred MS+NAA0.1mg/L of the composition of described root media.
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| CN103444536B (en) * | 2013-09-07 | 2015-07-01 | 河北省林业科学研究院 | Method for reducing tissue culture production cost of hosta plantagineu |
| 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 |
| CN103766219B (en) * | 2014-01-20 | 2016-08-17 | 上海上房园艺有限公司 | A kind of method of tissue culture long fringe torch flower |
| CN104303745B (en) * | 2014-09-24 | 2016-04-13 | 江苏省林业科学研究院 | A kind of method of jade hairpin plantlet in vitro outside sprout-cultivating-bottle |
| CN104304035B (en) * | 2014-11-14 | 2016-06-15 | 黑龙江省科学院自然与生态研究所 | The tissue cultivation rapid breeding method of golden-rimmed jade hairpin |
| CN110972945A (en) * | 2019-12-19 | 2020-04-10 | 美尚生态景观股份有限公司 | Culture medium and culture method for tissue culture of plantain plantaginea |
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| 徐刚等.玉簪的组培快繁.《中国花卉园艺》.2008,(第22期),15-17. |
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| 高志慧.引种玉簪(Hosta)抗寒性及组培快繁研究.《东北林业大学硕士学位论文》.2009,正文第35至37页. * |
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