CN104082152A - Tissue culture and rapid propagation method for clematis ranunculoides - Google Patents
Tissue culture and rapid propagation method for clematis ranunculoides Download PDFInfo
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- CN104082152A CN104082152A CN201410379137.2A CN201410379137A CN104082152A CN 104082152 A CN104082152 A CN 104082152A CN 201410379137 A CN201410379137 A CN 201410379137A CN 104082152 A CN104082152 A CN 104082152A
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Abstract
The invention provides a tissue culture and rapid propagation method for clematis ranunculoides. A clematis ranunculoides plant is finally obtained by acquisition of sterile raw materials, induction of buds, multiplication culture of adventitious buds, rooting culture and seedling exercising and transplantation of the plant. According to the method, the clematis ranunculoides can be rapidly propagated, the seedling culture period is short, the clematis ranunculoides plant can be rapidly obtained, the survival rate is higher than 90 percent, the propagation coefficients are high, propagation is not limited by external conditions and can be carried out in all seasons, the occupied area of seedling culture is saved, a great number of high-quality test-tube plantlets can be formed within a short time, and the clematis ranunculoides can be produced on a large scale.
Description
Technical field
The present invention relates to a kind of fast numerous method of plant tissue culture, especially a kind of method of buttercup clematis tissue-culturing rapid propagation, belongs to field of plant tissue culture technique.
Background technology
Buttercup clematis (
clematis ranunculoidesfranch
.) be Ranunculaceae herbaceous species plant, be distributed in Southwestern China area.Flower is intensive, aubergine, and September at florescence is to October, open on the occasion of the mid-autumn on National Day, has higher sight, can be used as pot flowers or climbers planting plant.Meanwhile, all herbal medicine, can treat various diseases.Therefore, buttercup clematis is to have the medicinal ornamental plants that exploitation is worth.
Yet in prior art, buttercup clematis mostly is self-sow and breeds, there is not yet the method for the relevant buttercup clematis of open report tissue-culturing rapid propagation.If will obtain efficiently, fast the buttercup clematis that this has medical value, be necessary to research and develop the technology of buttercup clematis tissue-culturing rapid propagation, short to realize buttercup clematis growing-seedling period, obtain fast buttercup clematis plant, keep the advantages such as maternal merit simultaneously.
Summary of the invention
Long for solving natural breed cycle of buttercup clematis, be difficult to adapt to medicinal demand, the invention provides a kind of method of buttercup clematis tissue-culturing rapid propagation, short to realize the growing-seedling period of buttercup clematis, make people can obtain fast buttercup clematis plant.
The present invention completes by following technical proposal: a kind of method of buttercup clematis tissue-culturing rapid propagation, is characterized in that through following each step:
(1) cut the tender shoots of buttercup clematis, water rinses 1h, alcohol immersion 25～the 35s that is 75% by volumetric concentration successively again, the mercuric chloride that is 1 ‰ by concentration soak 5～10min, then with after aseptic water washing 3～5 times, blot surface moisture, get tender shoots base portion and be cut into 1cm, be inoculated on bud inducing culture: MS+6-BA 1.5～2.5mg/L+NAA 0.4～0.6mg/L+AgNO
32.5～3.5mg/L is that 22～26 ℃, intensity of illumination are to cultivate under 1500～2000Lx, the relative moisture condition that is 65～75% in temperature;
(2) when the bastem portion of step (1) tender shoots starts to expand and occurs yellow green projection, then continue to cultivate after 20～25 days and occur obvious callus, then continue to cultivate 30～35 days, until callus grows budlet;
(3) cutting step (2) is inoculated on adventitious bud proliferation medium with the callus of bud: MS+BA 1.5～2.5mg/L+NAA 0.04～0.06mg/L, in temperature, be that 22～26 ℃, intensity of illumination are to cultivate under 1500～2000Lx, the relative moisture condition that is 65～75%, make adventitious bud proliferation, until indefinite bud grows to 2～3cm;
(4) getting step (3) height is the adventitious bud plants of 2～3cm, robust growth, transferred in root media: 1/2MS+IBA 0.4～0.6mg/L+NAA 0.9～1.1mg/L+AC 0.1～0.25g/L, in temperature, be that 22～26 ℃, intensity of illumination are to carry out root induction under 1500～2000Lx, the relative moisture condition that is 65～75%, after 30 days, the base section of plant seedling dissolves the former base of white root, until the former base of root grows to 3～5cm, obtain the seedling of taking root;
(5) bottle seedling of taking root of getting step (4) robust growth is opened sealed membrane transition 3～8 days in indoor, takes out afterwash root, transplants and in booth, tames 25～35 days, and then transplant extremely outdoor and give suitable rich water quality management, obtains buttercup clematis plant.
The present invention compared with prior art has following advantages and effect: this method energy Fast-propagation buttercup clematis; growing-seedling period is short; can obtain fast buttercup clematis plant; survival rate is up to more than 90%, and reproduction coefficient is high, and is not subject to the restriction of external condition; the four seasons all can carry out; the saving floor space of growing seedlings, can form a large amount of good test-tube plantlets in a short time, can carry out large-scale production.
Embodiment
Below in conjunction with embodiment, the present invention is described further.
Embodiment 1
(1) cut the tender shoots of buttercup clematis, water rinses 1h, the alcohol immersion 25s that is 75% by volumetric concentration successively on superclean bench, the mercuric chloride that is 1 ‰ by concentration soak 5min, then with after aseptic water washing 3 times, blot surface moisture, get tender shoots base portion and be cut into 1cm, pollution rate is 92%, is inoculated on bud inducing culture: MS+6-BA 1.5mg/L+NAA 0.5mg/L+AgNO
33mg/L is that 24 ℃, intensity of illumination are to cultivate under 1500Lx, the relative moisture condition that is 70% in temperature;
(2) when the bastem portion of step (1) tender shoots starts to expand and occurs yellow green projection, then continue to cultivate after 20 days and occur obvious callus, then continue to cultivate 30 days, until callus grows budlet, inductivity 64%;
(3) cutting step (2) is inoculated on adventitious bud proliferation medium with the callus of bud: MS+BA 1.5mg/L+NAA 0.05mg/L, in temperature, be that 26 ℃, intensity of illumination are to cultivate under 1800Lx, the relative moisture condition that is 75%, make adventitious bud proliferation, until indefinite bud grows to 2～3cm, the rate of increase of 20d is 6.8 times;
(4) getting step (3) height is the adventitious bud plants of 2～3cm, robust growth, transferred in root media: 1/2MS+IBA 0.5mg/L+NAA 1mg/L+AC 0.1g/L, in temperature, be that 26 ℃, intensity of illumination are to carry out root induction under 1500x, the relative moisture condition that is 65%, after 30 days, the base section of plant seedling dissolves the former base of white root, until the former base of root grows to 3～5cm, obtain the seedling of taking root, rooting rate 35%;
(5) bottle seedling of taking root of getting step (4) robust growth is opened sealed membrane transition 5 days in indoor, take out afterwash root, transplant and in plastic tunnel, tame 30 days, and then transplant to outdoor and give suitable rich water quality management, obtain buttercup clematis plant, survival rate 92%.
Embodiment 2
(1) cut the tender shoots of buttercup clematis, water rinses 1h, the alcohol immersion 30s that is 75% by volumetric concentration successively again, the mercuric chloride that is 1 ‰ by concentration soak 8min, then with after aseptic water washing 4 times, blot surface moisture, get tender shoots base portion and be cut into 1cm, pollution rate is 32.5%, is inoculated on bud inducing culture: MS+6-BA 2mg/L+NAA 0.4mg/L+AgNO
33.5mg/L is that 22 ℃, intensity of illumination are to cultivate under 1700Lx, the relative moisture condition that is 65% in temperature;
(2) when the bastem portion of step (1) tender shoots starts to expand and occurs yellow green projection, then continue to cultivate after 22 days and occur obvious callus, then continue to cultivate 33 days, until callus grows budlet, inductivity 83%;
(3) cutting step (2) is inoculated on adventitious bud proliferation medium with the callus of bud: MS+BA 2mg/L+NAA 0.04mg/L, in temperature, be that 24 ℃, intensity of illumination are to cultivate under 1500Lx, the relative moisture condition that is 70%, make adventitious bud proliferation, until indefinite bud grows to 2～3cm, the rate of increase of 20d is 11.5 times;
(4) getting step (3) height is the adventitious bud plants of 2～3cm, robust growth, transferred in root media: 1/2MS+IBA 0.4mg/L+NAA 1.1mg/L+AC 0.15g/L, in temperature, be that 24 ℃, intensity of illumination are to carry out root induction under 1700Lx, the relative moisture condition that is 70%, after 30 days, the base section of plant seedling dissolves the former base of white root, until the former base of root grows to 3～5cm, obtain the seedling of taking root, rooting rate 60%;
(5) bottle seedling of taking root of getting step (4) robust growth is opened sealed membrane transition 3 days in indoor, take out afterwash root, transplant and in plastic tunnel, tame 35 days, and then transplant to outdoor and give suitable rich water quality management, obtain buttercup clematis plant, survival rate 90.2%.
Embodiment 3
(1) cut the tender shoots of buttercup clematis, water rinses 1h, the alcohol immersion 35s that is 75% by volumetric concentration successively again, the mercuric chloride that is 1 ‰ by concentration soak 10min, then with after aseptic water washing 5 times, blot surface moisture, get tender shoots base portion and be cut into 1cm, pollution rate is 0, is inoculated on bud inducing culture: MS+6-BA 2.5mg/L+NAA 0.6mg/L+AgNO
32.5mg/L is that 26 ℃, intensity of illumination are to cultivate under 2000Lx, the relative moisture condition that is 75% in temperature;
(2) when the bastem portion of step (1) tender shoots starts to expand and occurs yellow green projection, then continue to cultivate after 25 days and occur obvious callus, then continue to cultivate 35 days, until callus grows budlet, inductivity 53%;
(3) cutting step (2) is inoculated on adventitious bud proliferation medium with the callus of bud: MS+BA 2.5mg/L+NAA 0.06mg/L, in temperature, be that 22 ℃, intensity of illumination are to cultivate under 2000Lx, the relative moisture condition that is 65%, make adventitious bud proliferation, until indefinite bud grows to 2～3cm, the rate of increase of 20 days is 5.1 times;
(4) getting step (3) height is the adventitious bud plants of 2～3cm, robust growth, transferred in root media: 1/2MS+IBA 0.6mg/L+NAA 0.9mg/L+AC 0.25g/L, in temperature, be that 22 ℃, intensity of illumination are to carry out root induction under 2000Lx, the relative moisture condition that is 75%, after 30 days, the base section of plant seedling dissolves the former base of white root, until the former base of root grows to 3～5cm, obtain the seedling of taking root, rooting rate 78.3%;
(5) bottle seedling of taking root of getting step (4) robust growth is opened sealed membrane transition 8 days in indoor, take out afterwash root, transplant and in plastic tunnel, tame 25 days, and then transplant to outdoor and give suitable rich water quality management, obtain buttercup clematis plant, survival rate 95.2%.
Claims (1)
1. a method for buttercup clematis tissue-culturing rapid propagation, is characterized in that through following each step:
(1) cut the tender shoots of buttercup clematis, water rinses 1h, alcohol immersion 25～the 35s that is 75% by volumetric concentration successively again, the mercuric chloride that is 1 ‰ by concentration soak 5～10min, then with after aseptic water washing 3～5 times, blot surface moisture, get tender shoots base portion and be cut into 1cm, be inoculated on bud inducing culture: MS+6-BA 1.5～2.5mg/L+NAA 0.4～0.6mg/L+AgNO
32.5～3.5mg/L is that 22～26 ℃, intensity of illumination are to cultivate under 1500～2000Lx, the relative moisture condition that is 65～75% in temperature;
(2) when the bastem portion of step (1) tender shoots starts to expand and occurs yellow green projection, then continue to cultivate after 20～25 days and occur obvious callus, then continue to cultivate 30～35 days, until callus grows budlet;
(3) cutting step (2) is inoculated on adventitious bud proliferation medium with the callus of bud: MS+BA 1.5～2.5mg/L+NAA 0.04～0.06mg/L, in temperature, be that 22～26 ℃, intensity of illumination are to cultivate under 1500～2000Lx, the relative moisture condition that is 65～75%, make adventitious bud proliferation, until indefinite bud grows to 2～3cm;
(4) getting step (3) height is the adventitious bud plants of 2～3cm, robust growth, transferred in root media: 1/2MS+IBA 0.4～0.6mg/L+NAA 0.9～1.1mg/L+AC 0.1～0.25g/L, in temperature, be that 22～26 ℃, intensity of illumination are to carry out root induction under 1500～2000Lx, the relative moisture condition that is 65～75%, after 30 days, the base section of plant seedling dissolves the former base of white root, until the former base of root grows to 3～5cm, obtain the seedling of taking root;
(5) bottle seedling of taking root of getting step (4) robust growth is opened sealed membrane transition 3～8 days in indoor, takes out afterwash root, transplants and in booth, tames 25～35 days, and then transplant extremely outdoor and give suitable rich water quality management, obtains buttercup clematis plant.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106134994A (en) * | 2016-06-29 | 2016-11-23 | 无锡南理工科技发展有限公司 | A kind of artificial cultivation method of radix ranunculi ternati |
| CN107567760A (en) * | 2017-09-29 | 2018-01-12 | 江苏农林职业技术学院 | A kind of method for improving long flower clematis percentage of seedgermination and emergence rate |
| CN107580822A (en) * | 2017-09-29 | 2018-01-16 | 江苏农林职业技术学院 | A kind of method for improving bastard clematis percentage of seedgermination and emergence rate |
| CN108124749A (en) * | 2017-12-12 | 2018-06-08 | 浙江省亚热带作物研究所 | A kind of method that clematis Water culture is taken root |
| CN108184662A (en) * | 2016-12-08 | 2018-06-22 | 上海植物园 | The high-efficiency in-vitro quick-breeding method and its culture medium of a kind of clematis |
| CN109601388A (en) * | 2019-01-29 | 2019-04-12 | 中国林业科学研究院林业研究所 | A kind of quick breeding method for tissue culture hybridizing clematis |
| CN113207687A (en) * | 2021-05-18 | 2021-08-06 | 西南林业大学 | Tissue culture and rapid propagation method for clematis |
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| NL194191B (en) * | 1994-01-13 | 2001-05-01 | Jan Hendrik Fondse | Method for growing a plant from a plant part. |
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106134994A (en) * | 2016-06-29 | 2016-11-23 | 无锡南理工科技发展有限公司 | A kind of artificial cultivation method of radix ranunculi ternati |
| CN108184662A (en) * | 2016-12-08 | 2018-06-22 | 上海植物园 | The high-efficiency in-vitro quick-breeding method and its culture medium of a kind of clematis |
| CN108184662B (en) * | 2016-12-08 | 2021-09-03 | 上海植物园 | Efficient in-vitro rapid propagation method and culture medium of clematis |
| CN107567760A (en) * | 2017-09-29 | 2018-01-12 | 江苏农林职业技术学院 | A kind of method for improving long flower clematis percentage of seedgermination and emergence rate |
| CN107580822A (en) * | 2017-09-29 | 2018-01-16 | 江苏农林职业技术学院 | A kind of method for improving bastard clematis percentage of seedgermination and emergence rate |
| CN108124749A (en) * | 2017-12-12 | 2018-06-08 | 浙江省亚热带作物研究所 | A kind of method that clematis Water culture is taken root |
| CN108124749B (en) * | 2017-12-12 | 2019-11-05 | 浙江省亚热带作物研究所 | A kind of method that clematis Water culture is taken root |
| CN109601388A (en) * | 2019-01-29 | 2019-04-12 | 中国林业科学研究院林业研究所 | A kind of quick breeding method for tissue culture hybridizing clematis |
| CN109601388B (en) * | 2019-01-29 | 2022-03-22 | 中国林业科学研究院林业研究所 | Tissue culture rapid propagation method of hybrid clematis |
| CN113207687A (en) * | 2021-05-18 | 2021-08-06 | 西南林业大学 | Tissue culture and rapid propagation method for clematis |
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Application publication date: 20141008 |