CN104206281A - Tissue cultivation method for elegant-purple clematis - Google Patents
Tissue cultivation method for elegant-purple clematis Download PDFInfo
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- CN104206281A CN104206281A CN201410520037.7A CN201410520037A CN104206281A CN 104206281 A CN104206281 A CN 104206281A CN 201410520037 A CN201410520037 A CN 201410520037A CN 104206281 A CN104206281 A CN 104206281A
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Abstract
The invention discloses a tissue cultivation method for elegant-purple clematis. The method comprises the steps of: (1) obtaining an aseptic seeding material; (2) performing primary culture; (3) performing enrichment culture; and (4) performing rooting and seedling strengthening culture; when the root length reaches 2-4cm, acclimating and transplanting can be performed. The tissue cultivation method for elegant-purple clematis provided by the invention has the advantages that, propagation cycle is short, propagation coefficient is high, roots are easy to grow, root systems of plants are thick and strong, growth vigor of new shoots is good, stalks are thick and strong, and acclimated seedling is easy to survive.
Description
Technical field
The present invention relates to a kind of ornamental plant compound formulation, especially relate to the tissue culture method of a kind of clematis, belong to propagation technique field.
Background technology
Clematis is subordinate to Ranunculaceae (clematis fusca var.violacea) Clematis (Clematis L.), is the liane that a class ornamental value is high, have multiple resistance.Actions of Clematis Species is perennial wooden or herbaceous species, and minority is upright shrub.Leaf, to life, has handle.Dan Ye, trifoliolate leaf or winglike compound leaf, Quan Yuan, have sawtooth or division.Cyme, panicle or Dan Sheng.Actions of Clematis Species flower shape is new and original, and change is large, and flower color and luster is in harmonious proportion, unique elegance; Florescence is longer, strong adaptability, and has stronger cold resistance, in international ornamental horticulture, occupying critical role.In today that pursuit flower variety is new, strange, special, the ornamental value of Actions of Clematis Species is more and more subject to the attention of domestic and international gardening circle.Clematis seminal propagation rate is very low, cottage propagation rooting rate is not high yet, tissue cultures and body embryo is utilized to be the approach having application prospect, a lot of plant (as ivyleaf cyclamen) utilizes tissue cultures to be produced on a large scale, and it is less to the tissue cultures studies in China of clematis, development speed relatively lags behind, and has had a strong impact on the industrialized development of excellent clematis kind.Therefore carrying out Study on tissue culture to clematis, explore form development ways and the isolated culture condition of high-frequency regeneration plant, be used to guide seeling industry, simultaneously also for research Actions of Clematis Species Regeneration in Vitro rule provides some data, is very necessary.
Summary of the invention
For solving the deficiencies in the prior art, the object of the present invention is to provide a kind of breeding cycle short, reproduction coefficient is high and be easy to a kind of clematis Viticella G. ' Purpurea Plena Elegans ' method for tissue culture of taking root.
In order to solve above-mentioned purpose, the technical solution adopted in the present invention is:
A kind of method for tissue culture of clematis Viticella G. ' Purpurea Plena Elegans ' (graceful purple), comprises the following steps:
(1) aseptic seedling material is obtained:
The stem with bud that clip clematis newly sprouts spring, to rinse after 30-60min under running water on superclean bench, with the alcohol immersion 15-20s of volumetric concentration 70-75%, the mercuric chloride solution of 1% (mass concentration) is used to soak after 8-15 minute again, with aseptic water washing 5-8 time, blot stem with bud surface moisture;
(2) Initial culture:
Get the stem with bud after above-mentioned aseptic process, be cut into the segment that 1cm is long, every section is at least with one piece of growth full, the healthy bud of anosis worm harm shape, be inoculated on inducing culture that pH value is 5.6-6.0, described stem with bud starts to produce Multiple Buds for 7 days after inducing culture Fiber differentiation, continues cultivation after 4 weeks, and the shoot proliferation carrying out step (3) is cultivated;
(3) Multiplying culture:
Be inoculated on adventitious bud proliferation medium that pH value is 5.8-6.2 with the clematis of Multiple Buds and carry out Multiplying culture, increment cultivation 30 days, illumination every day 16h, Multiplying culture temperature is 24-26 DEG C, and intensity of illumination is 30-40 μm of ol/m
2s;
(4) Rooting and hardening-off culture:
Induce Multiple Buds in adventitious bud proliferation medium in step (3) after, Multiple Buds is cut into the stem section of individual plant or 2.0-3.5cm length, is inoculated in the Rooting and hardening-off culture base of pH5.6-6.0, after light culture 7-10 days, carries out illumination cultivation; When root length is for 2-4cm, rooting culture can be carried out.
Further, in step (2), described Fiber differentiation based component is 1/2MS+0.2mg/L6-BA+0.5mg/LKT+0.05mg/LNAA+3% (mass concentration) sucrose+0.7% (mass concentration) agar; PH value is 5.8.
Further, in step (3), described Multiplying culture based component is 1/2MS+0.2mg/L6-BA+0.5mg/LKT+0.05mg/LNAA+3% (mass concentration) sucrose+0.7% (mass concentration) agar+10% (mass concentration) Coconut Juice+0.1mgVc.
Further, in step (4), described culture of rootage based component is 1/3MS+NAA0.1-0.5mg.L
-1+ 0.5g active carbon.
Beneficial effect: compared with prior art, clematis method for tissue culture provided by the present invention, the breeding cycle is short, and reproduction coefficient is high, easily takes root, and the root system of plant is sturdy, and newly slightly growing way is good, and stem stalk is sturdy, and domestication easily survives.
Embodiment:
Below will the present invention will be described according to specific embodiment:
In the present embodiment, the basic media components MS be applied in each medium all adopts identical formula, and wherein, the formula of MS is as follows:
Macroelement (mg/l) comprising:
Trace element (mg/l) comprising:
Molysite (mg/l) comprising:
FeSO
4·7H
2O 27.8
Na
2-EDTA·2H
2O 37.3；
Organic substance (mg/l) comprising:
6-BA:6-benzyl aminoadenine;
KT: kinetin;
NAA: methyl α-naphthyl acetate;
Vc: vitamin C.
Clematis kind used in the present invention is clematis Viticella G. ' Purpurea Plena Elegans ' (graceful purple).
Embodiment 1:
A kind of clematis method for tissue culture, is characterized in that, comprise the following steps:
(1) aseptic seedling material is obtained:
The stem with bud that clip clematis newly sprouts spring, to rinse after 30min under running water on superclean bench, with the alcohol immersion 15s of volumetric concentration 70%, after using the mercuric chloride solution of 1% (mass concentration) to soak 8 minutes again, with aseptic water washing 5 times, blot stem with bud surface moisture;
(2) Initial culture:
Get the stem with bud after above-mentioned aseptic process, be cut into the segment that 1cm is long, every section is at least with one piece of growth full, the healthy bud of anosis worm harm shape, being inoculated in pH value is on the inducing culture of 5.8, described stem with bud starts to produce Multiple Buds for 7 days after inducing culture Fiber differentiation, continues cultivation after 4 weeks, and the shoot proliferation carrying out step (3) is cultivated; Wherein, Fiber differentiation based component is 1/2MS+0.2mg/L6-BA+0.5mg/LKT+0.05mg/LNAA+3% (mass concentration) sucrose+0.7% (mass concentration) agar; PH value is 5.8.
(3) Multiplying culture:
Be inoculated on adventitious bud proliferation medium that pH value is 5.8-6.2 with the clematis of Multiple Buds and carry out Multiplying culture, increment cultivation 30 days, illumination every day 16h, Multiplying culture temperature is 24-26 DEG C, and intensity of illumination is 30-40 μm of ol/m
2s; Wherein, Multiplying culture based component is 1/2MS+0.2mg/L6-BA+0.5mg/LKT+0.05mg/LNAA+3% (mass concentration) sucrose+0.7% (mass concentration) agar+10% (volumetric concentration) Coconut Juice+0.1mgVc, and pH value is 5.8-6.2.
(4) Rooting and hardening-off culture:
Induce Multiple Buds in adventitious bud proliferation medium in step (3) after, Multiple Buds is cut into the stem section of individual plant or 2.0-3.5cm length, be inoculated in the Rooting and hardening-off culture base of pH5.6-6.0, light culture carried out illumination cultivation after 7 days; When root length is for 2-4cm, rooting culture can be carried out; Wherein, culture of rootage based component is 1/3MS+NAA0.1-0.5mg.L
-1+ 0.5g active carbon, pH5.6-6.0.
Embodiment 2: a kind of clematis method for tissue culture, is characterized in that, comprise the following steps:
(1) aseptic seedling material is obtained:
The stem with bud that clip clematis newly sprouts spring, to rinse after 60min under running water on superclean bench, with the alcohol immersion 20s of (volumetric concentration) 75%, after using the mercuric chloride solution of 1% (mass concentration) to soak 15 minutes again, with aseptic water washing 8 times, blot stem with bud surface moisture;
(2) Initial culture:
Get the stem with bud after above-mentioned aseptic process, be cut into the segment that 1cm is long, every section is at least with one piece of growth full, the healthy bud of anosis worm harm shape, being inoculated in pH value is on the inducing culture of 5.6, described stem with bud starts to produce Multiple Buds for 7 days after inducing culture Fiber differentiation, continues cultivation after 4 weeks, and the shoot proliferation carrying out step (3) is cultivated; Wherein, Fiber differentiation based component is 1/2MS+0.2mg/L6-BA+0.5mg/LKT+0.05mg/LNAA+3% (mass concentration) sucrose+0.7% (mass concentration) agar; PH value is 5.6.
(3) Multiplying culture:
Be inoculated on adventitious bud proliferation medium that pH value is 5.8-6.2 with the clematis of Multiple Buds and carry out Multiplying culture, 30d days, illumination every day 16h are cultivated in increment, and Multiplying culture temperature is 24-26 DEG C, and intensity of illumination is 30-40 μm of ol/m
2s; Wherein, Multiplying culture based component is 1/2MS+0.2mg/L6-BA+0.5mg/LKT+0.05mg/LNAA+3% (mass concentration) sucrose+0.7% (mass concentration) agar+10% (mass concentration) Coconut Juice+0.1mgVc.
(4) Rooting and hardening-off culture:
Induce Multiple Buds in adventitious bud proliferation medium in step (3) after, Multiple Buds is cut into the stem section of individual plant or 2.0-3.5cm length, be inoculated in the Rooting and hardening-off culture base of pH5.6-6.0, light culture carried out illumination cultivation after 10 days; When root length is for 2-4cm, rooting culture can be carried out; Wherein, culture of rootage based component is 1/3MS+NAA0.1-0.5mg.L
-1+ 0.5g active carbon.
The result obtained after cultivating the graceful purple of clematis according to the method for above-described embodiment 1 and embodiment 2 is as shown in the table:
| Reproduction coefficient | Rooting rate | Survival rate | |
| Embodiment 1 | 3.2 | 98.2％ | 88.2％ |
| Embodiment 2 | 3.4 | 97.8％ | 90.1％ |
The present invention is illustrated according to above-described embodiment and should be appreciated that above-described embodiment does not limit the present invention in any form, and all employings are equal to replacement or the technical scheme that obtains of equivalent transformation mode, all drop within protection scope of the present invention.
Claims (5)
1. the graceful purple method for tissue culture of clematis, is characterized in that, comprise the following steps:
(1) aseptic seedling material is obtained:
The stem with bud that clip clematis newly sprouts spring, to rinse after 30-60min under running water on superclean bench, with the alcohol immersion 15-20s of volumetric concentration 70-75%, after soaking 8-15 minute with the mercuric chloride solution of mass concentration 1% again, with aseptic water washing 5-8 time, blot stem with bud surface moisture;
(2) Initial culture:
Get the stem with bud after above-mentioned aseptic process, be cut into the segment that 1cm is long, every section is at least with one piece of growth full, the healthy bud of anosis worm harm shape, be inoculated on inducing culture that pH value is 5.6-6.0, described stem with bud starts to produce Multiple Buds for 7 days after inducing culture Fiber differentiation, continues cultivation after 4 weeks, and the shoot proliferation carrying out step (3) is cultivated;
(3) Multiplying culture:
Be inoculated on adventitious bud proliferation medium that pH value is 5.8-6.2 with the clematis of Multiple Buds and carry out Multiplying culture, increment cultivation 30 days, illumination every day 16h, Multiplying culture temperature is 24-26 DEG C, and intensity of illumination is 30-40 μm of ol/m
2s;
(4) Rooting and hardening-off culture:
Induce Multiple Buds in adventitious bud proliferation medium in step (3) after, Multiple Buds is cut into the stem section of individual plant or 2.0-3.5cm length, is inoculated in the Rooting and hardening-off culture base of pH5.6-6.0, after light culture 7-10 days, carries out illumination cultivation; When root length is for 2-4cm, rooting culture can be carried out.
2. the graceful purple method for tissue culture of clematis according to claim 1, it is characterized in that, described Fiber differentiation based component is 1/2MS+0.2mg/L6-BA+0.5mg/LKT+0.05mg/LNAA+3% (mass concentration) sucrose+0.7% (mass concentration) agar; PH value is 5.8.
3. the graceful purple method for tissue culture of clematis according to claim 1, it is characterized in that, described Multiplying culture based component is 1/2MS+0.2mg/L6-BA+0.5mg/LKT+0.05mg/LNAA+3% (mass concentration) sucrose+0.7% (mass concentration) agar+10% (mass concentration) Coconut Juice+0.1mgVc.
4. the graceful purple method for tissue culture of clematis according to claim 1, it is characterized in that, described culture of rootage based component is 1/3MS+NAA0.1-0.5mg.L
-1+ 0.5g active carbon.
5. the graceful purple method for tissue culture of the clematis according to any one of claim 2-4 claim, it is characterized in that, wherein described, the formula of MS is as follows:
Macroelement (mg/l) comprising:
Trace element (mg/l) comprising:
Molysite (mg/l) comprising:
FeSO
4·7H
2O 27.8
Na
2-EDTA·2H
2O 37.3；
Organic substance (mg/l) comprising:
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Cited By (7)
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| CN105660400A (en) * | 2016-01-25 | 2016-06-15 | 宁波易中禾生物技术有限公司 | Strengthening and weight increasing method for anoectochilus roxburghii tissue cultured seedlings |
| CN107041307A (en) * | 2017-04-26 | 2017-08-15 | 江苏农林职业技术学院 | Clematis Henry method for tissue culture |
| CN109618805A (en) * | 2019-02-27 | 2019-04-16 | 湖南春光九汇现代中药有限公司 | A kind of implantation methods of evodia rutaecarpa |
| CN109618927A (en) * | 2018-12-21 | 2019-04-16 | 滁州学院 | A kind of method for tissue culture of sweetautumn clematis |
| CN110547202A (en) * | 2019-10-09 | 2019-12-10 | 江苏农林职业技术学院 | clematis chinensis proliferation tissue culture method |
| CN110637722A (en) * | 2019-10-24 | 2020-01-03 | 江苏农林职业技术学院 | Induction method of clematis warrior dance callus |
| CN114391475A (en) * | 2022-01-26 | 2022-04-26 | 江苏农林职业技术学院 | Tissue culture method of clematis krispipe angel |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105660400A (en) * | 2016-01-25 | 2016-06-15 | 宁波易中禾生物技术有限公司 | Strengthening and weight increasing method for anoectochilus roxburghii tissue cultured seedlings |
| CN107041307A (en) * | 2017-04-26 | 2017-08-15 | 江苏农林职业技术学院 | Clematis Henry method for tissue culture |
| CN109618927A (en) * | 2018-12-21 | 2019-04-16 | 滁州学院 | A kind of method for tissue culture of sweetautumn clematis |
| CN109618805A (en) * | 2019-02-27 | 2019-04-16 | 湖南春光九汇现代中药有限公司 | A kind of implantation methods of evodia rutaecarpa |
| CN110547202A (en) * | 2019-10-09 | 2019-12-10 | 江苏农林职业技术学院 | clematis chinensis proliferation tissue culture method |
| CN110637722A (en) * | 2019-10-24 | 2020-01-03 | 江苏农林职业技术学院 | Induction method of clematis warrior dance callus |
| CN114391475A (en) * | 2022-01-26 | 2022-04-26 | 江苏农林职业技术学院 | Tissue culture method of clematis krispipe angel |
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