CN104705186A - Tissue culture and quick propagation method of S. versicolor - Google Patents
Tissue culture and quick propagation method of S. versicolor Download PDFInfo
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- CN104705186A CN104705186A CN201510103910.7A CN201510103910A CN104705186A CN 104705186 A CN104705186 A CN 104705186A CN 201510103910 A CN201510103910 A CN 201510103910A CN 104705186 A CN104705186 A CN 104705186A
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Abstract
The invention discloses a tissue culture and quick propagation method of S. versicolor. The S. versicolor is an excellent ornamental bamboo species in the grass family. Massive and quick propagation can be carried out on a plant by using a plant tissue culture technology, the plant better adapts to an existing environment, and an ornamental property of the plant can be changed at the same time. The method takes a stem, with a bud, of the S. versicolor as an explant; an in-vitro replant of the S. versicolor is obtained by processes such as explant disinfection, induction culture, subculture, rooting culture, acclimatization and transplanting; a tissue culture and quick propagation technical system of the S. versicolor is established; and the method has an important significance to promote large-scale development of the S. versicolor.
Description
Technical field
The present invention relates to the method for Plant Tissue Breeding in agricultural biotechnologies, specifically, relate to a kind of daybreak Ce bamboo tissue culture rapid breeding method.
Background technology
Eighties of last century sixties plant tissue culture technique is day by day ripe, and starts to obtain large-scale application in production.For the rare bamboo kind having higher ornamental value, Sterile culture is difficult to meet society need, and employing tube rapid propagation can large-scale production bamboo seedling.The tissue cultures of bamboo plant is started in 1968, and the 80's abroad started to compare systematic research.Ce Sinobambusa, Phyllostachys, Dendrocalamus, cold Sinobambusa, Indocalamus Nakai etc. more than 20 are belonged to more than 70 bamboo kind and carried out the research of tissue cultures, and mainly concentrate on Cluster Bamboo kind, about the reported success of Scattered bamboos kind tissue cultures and tube rapid propagation is less so far.
Daybreak Ce bamboo (
s. versicolor) be grass family bamboo subgenus, originate in Japan.For excellent ornament bamboo, do not resist cold.Onset young leaves white, transfers floral leaf to subsequently, finally becomes entirely green, have very high ornamental value.Bamboo due to routine breed not only efficiency low, expend large, the problem of bamboo industry seedling demand cannot be solved.Utilize plant tissue culture technique, can carry out a large amount of to plant and breed fast, the foundation of the regenerating system of bamboo has important basic role to its Study on Genetic Transformation, the genetic character of plant is changed by genetic transformation, make it better adapt to the environment of existence, can change it views and admires character simultaneously.Therefore; the present invention with daybreak Ce bamboo stem with bud for explant; the in vitro plant again of daybreak Ce bamboo is obtained by processes such as explant sterilization, Fiber differentiation, squamous subculture, culture of rootage, acclimatization and transplantses; set up daybreak Ce bamboo tissue culture rapid propagation technique system, significant to promotion daybreak Ce bamboo large-scale development.
Summary of the invention
The object of the present invention is to provide out a kind of daybreak Ce bamboo tissue culture rapid breeding method, the present invention with daybreak Ce bamboo stem with bud for explant, the in vitro plant again of daybreak Ce bamboo is obtained by processes such as explant sterilization, Fiber differentiation, squamous subculture, culture of rootage, acclimatization and transplantses, set up daybreak Ce bamboo tissue culture rapid propagation technique system, thus achieve object of the present invention.
A kind of daybreak Ce bamboo tissue culture rapid breeding method of the present invention, comprises the following steps:
(1) explant sterilization: choose daybreak Ce bamboo green tape leaf stem section then, rinses 1 ~ 3h under flowing water, with banister brush 3% ~ 5% washing powder solution washing material gently, then rinses 60 ~ 90min with running water with the form of dripping.With 70% ~ 80% alcohol solution dipping 20 ~ 60s in superclean bench, aseptic water washing 3 ~ 5 times, then with 0.1% ~ 0.5% mercuric chloride solution sterilization 5 ~ 15min, blots the moisture on surface with aseptic filter paper after aseptic water washing 3 ~ 5 times for subsequent use.
(2) Fiber differentiation: be inoculated into inducing culture after the stem section after step (1) process is peelled off stalk sheaths of bamboo shoots and carry out inducing clumping bud cultivation.Inoculation is placed on illumination every day 12 ~ 14 hours, and intensity of illumination is 2000 ~ 3000lx, and being placed in cultivation temperature is 25 ~ 28 DEG C, and relative air humidity is cultivate under the condition of 75% ~ 80% to add up induction situation afterwards in 30 days.
(3) Multiplying culture: step (2) Fiber differentiation being obtained length is that the aseptic bud of about 1.5 ~ 2.5cm growing way is inoculated into proliferated culture medium and carries out adventitious buds proliferation cultivation.Inoculation is placed on illumination every day 12 ~ 14 hours, and intensity of illumination is 2000 ~ 3000lx, and being placed in cultivation temperature is 25 ~ 28 DEG C, and relative air humidity is cultivate under the condition of 75% ~ 80% after 30 days to add up germinative number.
(4) culture of rootage: cut in step (3) propagation and obtain the seedling of high about 3.5 ~ 4.5cm and be seeded in root media and carry out root induction.Inoculation is placed on illumination every day 12 ~ 14 hours, and intensity of illumination is 2000 ~ 3000lx, and being placed in cultivation temperature is 25 ~ 28 DEG C, and relative air humidity is cultivate under the condition of 75% ~ 80% to add up situation of taking root afterwards in 30 days.
(5) acclimatization and transplants: the bottle seedling possessing transplanting condition that root media obtains is carried out hardening, bottle seedling to move under normal temperature after 4 ~ 5 days, open bottle cap 2 ~ 3 days, then washing away the medium being attached to shoot root and fastening, transplanting to filling by peat soil: perlite: cultivate in the container bag of the transplanting culture matrix of vermiculite=1:1:1, each container bag is transplanted 1 strain and to be taken root seedling, need by matrix compacting during transplanting, and carry out individual plant bagging, spraying sooner or later, ensure that growing environment is moist.Transplant and cut off plastic sack two jiaos after 7 days, after 14 days, completely de-bag, transplants and adds up survival rate after 30 days.
Inducing culture described in above-mentioned steps (2) is: MS+3.0 ~ 8.0mg/L6-BA+0.1 ~ 1.0mg/L ZT+1.0 ~ 3.0g/L AC+ 15 ~ 30g/L sucrose+3.5 ~ 6.0g/L agar, pH is 5.4 ~ 5.8.
Proliferated culture medium described in above-mentioned steps (3) is: MS+0.001 ~ 0.01mg/L TDZ+1.0 ~ 3.0mg/L ZT+1.0 ~ 4.0mg/L 6-BA+15 ~ 30g/L sucrose+3.5 ~ 6.0g/L agar, pH is 5.4 ~ 5.8.
Root media described in above-mentioned steps (4) is: MS+0.1 ~ 1.0mg/L IBA+1.0 ~ 3.0mg/L IAA+15 ~ 30g/L sucrose+3.5 ~ 6.0g/L agar, pH is 5.4 ~ 5.8.
Compared with prior art advantage of the present invention is: daybreak Ce bamboo (
s. versicolor) be excellent ornament bamboo gramineous.Bamboo due to routine breed not only efficiency low, expend large, the problem of bamboo industry seedling demand cannot be solved.Utilize plant tissue culture technique, can carry out a large amount of to plant and breed fast, the foundation of the regenerating system of bamboo has important basic role to its Study on Genetic Transformation, the genetic character of plant is changed by genetic transformation, make it better adapt to the environment of existence, can change it views and admires character simultaneously.The present invention with daybreak Ce bamboo stem with bud for explant; the in vitro plant again of daybreak Ce bamboo is obtained by processes such as explant sterilization, Fiber differentiation, squamous subculture, culture of rootage, acclimatization and transplantses; set up daybreak Ce bamboo tissue culture rapid propagation technique system, significant to promotion daybreak Ce bamboo large-scale development.
Embodiment
Following examples further illustrate of the present invention, is not limitation of the present invention.
embodiment 1
(1) explant sterilization: choose daybreak Ce bamboo green tape leaf stem section then, rinse 1h under flowing water, with banister brush 3% washing powder solution washing material gently, then rinses 60min with running water with the form of dripping.With 75% alcohol solution dipping 30s in superclean bench, aseptic water washing 3 ~ 5 times, then with 0.1% mercuric chloride solution sterilization 5min, blots the moisture on surface with aseptic filter paper after aseptic water washing 3 times for subsequent use.
(2) Fiber differentiation: be inoculated into inducing culture after the stem section after step (1) process is peelled off stalk sheaths of bamboo shoots and carry out inducing clumping bud cultivation.Inoculation is placed on illumination every day 12 hours, and intensity of illumination is 2000lx, and being placed in cultivation temperature is 25 DEG C, and relative air humidity is that to cultivate inductivity after 30 days under the condition of 75% be 78.92%.Described inducing culture is: MS+5.0mg/L6-BA+0.5mg/L ZT+1.5g/L AC+ 18g/L sucrose+4.5g/L agar, pH is 5.5.
(3) Multiplying culture: step (2) Fiber differentiation being obtained length is that the aseptic bud of about 1.5 ~ 2.5cm growing way is inoculated into proliferated culture medium and carries out adventitious buds proliferation cultivation.Inoculation is placed on illumination every day 13 hours, and intensity of illumination is 2000lx, and being placed in cultivation temperature is 25 DEG C, and relative air humidity is that under the condition of 75% ~ 80%, cultivation 30 days growth coefficients are 3.62.Described proliferated culture medium is: MS+0.005mg/L TDZ+2.0mg/L ZT+2.5mg/L 6-BA+18g/L sucrose+4.5g/L agar, pH is 5.5.
(4) culture of rootage: cut in step (3) propagation and obtain the seedling of high about 3.5 ~ 4.5cm and be seeded in root media and carry out root induction.Inoculation is placed on illumination every day 12 hours, and intensity of illumination is 2000lx, and being placed in cultivation temperature is 25 DEG C, and relative air humidity is that to cultivate rooting rate after 30 days under the condition of 75% be 73.9%.Described root media is: MS+0.3mg/L IBA+1.5mg/L IAA+18g/L sucrose+4.5g/L agar, pH is 5.5.
(5) acclimatization and transplants: the bottle seedling possessing transplanting condition that root media obtains is carried out hardening, bottle seedling moves to normal temperature after lower 4 days, open bottle cap 2 days, then washing away the medium being attached to shoot root and fastening, transplanting to filling by peat soil: perlite: cultivate in the container bag of the transplanting culture matrix of vermiculite=1:1:1, each container bag is transplanted 1 strain and to be taken root seedling, need by matrix compacting during transplanting, and carry out individual plant bagging, spraying sooner or later, ensure that growing environment is moist.Transplant and cut off plastic sack two jiaos after 7 days, completely de-bag after 14 days, transplanting survival rate after 30 days is 93.48%.
Embodiment 2
(1) explant sterilization: choose daybreak Ce bamboo green tape leaf stem section then, rinse 2h under flowing water, with banister brush 3% washing powder solution washing material gently, then rinses 75min with running water with the form of dripping.With 75% alcohol solution dipping 40s in superclean bench, aseptic water washing 4 times, then with 0.1% mercuric chloride solution sterilization 12min, blots the moisture on surface with aseptic filter paper after aseptic water washing 5 times for subsequent use.
(2) Fiber differentiation: be inoculated into inducing culture after the stem section after step (1) process is peelled off stalk sheaths of bamboo shoots and carry out inducing clumping bud cultivation.Inoculation is placed on illumination every day 14 hours, and intensity of illumination is 2500lx, and being placed in cultivation temperature is 28 DEG C, and relative air humidity is that to cultivate inductivity after 30 days under the condition of 75% be 81.15%.Described inducing culture is: MS+7.0mg/L6-BA+0.7mg/L ZT+1.0g/L AC+ 23g/L sucrose+4.8g/L agar, pH is 5.8.
(3) Multiplying culture: step (2) Fiber differentiation being obtained length is that the aseptic bud of about 1.5 ~ 2.5cm growing way is inoculated into proliferated culture medium and carries out adventitious buds proliferation cultivation.Inoculation is placed on illumination every day 14 hours, and intensity of illumination is 2500lx, and being placed in cultivation temperature is 28 DEG C, and relative air humidity is that under the condition of 75% ~ 80%, cultivation 30 days growth coefficients are 3.49.Described proliferated culture medium is: MS+0.01mg/L TDZ+3.0mg/L ZT+3.0mg/L 6-BA+18g/L sucrose+4.5g/L agar, pH is 5.8.
(4) culture of rootage: cut in step (3) propagation and obtain the seedling of high about 3.5 ~ 4.5cm and be seeded in root media and carry out root induction.Inoculation is placed on illumination every day 13 hours, and intensity of illumination is 2500lx, and being placed in cultivation temperature is 25 DEG C, and relative air humidity is that to cultivate rooting rate after 30 days under the condition of 75% be 84.7%.Described root media is: MS+0.5mg/L IBA+2.0mg/L IAA+20g/L sucrose+4.8g/L agar, pH is 5.8.
(5) acclimatization and transplants: the bottle seedling possessing transplanting condition that root media obtains is carried out hardening, bottle seedling moves to normal temperature after lower 5 days, open bottle cap 3 days, then washing away the medium being attached to shoot root and fastening, transplanting to filling by peat soil: perlite: cultivate in the container bag of the transplanting culture matrix of vermiculite=1:1:1, each container bag is transplanted 1 strain and to be taken root seedling, need by matrix compacting during transplanting, and carry out individual plant bagging, spraying sooner or later, ensure that growing environment is moist.Transplant and cut off plastic sack two jiaos after 7 days, completely de-bag after 14 days, transplanting survival rate after 30 days is 91.97%.
Claims (4)
1. a daybreak Ce bamboo tissue culture rapid breeding method, is characterized in that comprising the following steps:
(1) explant sterilization: choose daybreak Ce bamboo green tape leaf stem section then; 1 ~ 3h is rinsed under flowing water; with banister brush 3% ~ 5% washing powder solution washing material gently; 60 ~ 90min is rinsed with the form of dripping again with running water; with 70% ~ 80% alcohol solution dipping 20 ~ 60s in superclean bench; aseptic water washing 3 ~ 5 times, then with 0.1% ~ 0.5% mercuric chloride solution sterilization 5 ~ 15min, blots the moisture on surface with aseptic filter paper after aseptic water washing 3 ~ 5 times for subsequent use;
(2) Fiber differentiation: be inoculated into inducing culture after the stem section after step (1) process is peelled off stalk sheaths of bamboo shoots and carry out inducing clumping bud cultivation; inoculation is placed on illumination every day 12 ~ 14 hours; intensity of illumination is 2000 ~ 3000lx; being placed in cultivation temperature is 25 ~ 28 DEG C, and relative air humidity is cultivate under the condition of 75% ~ 80% to add up induction situation afterwards in 30 days;
(3) Multiplying culture: step (2) Fiber differentiation being obtained length is that the aseptic bud of about 1.5 ~ 2.5cm growing way is inoculated into proliferated culture medium and carries out adventitious buds proliferation cultivation; inoculation is placed on illumination every day 12 ~ 14 hours; intensity of illumination is 2000 ~ 3000lx; being placed in cultivation temperature is 25 ~ 28 DEG C, and relative air humidity is cultivate under the condition of 75% ~ 80% after 30 days to add up germinative number;
(4) culture of rootage: cut in step (3) propagation and obtain the seedling of high about 3.5 ~ 4.5cm and be seeded in root media and carry out root induction; inoculation is placed on illumination every day 12 ~ 14 hours; intensity of illumination is 2000 ~ 3000lx; being placed in cultivation temperature is 25 ~ 28 DEG C, and relative air humidity is cultivate under the condition of 75% ~ 80% to add up situation of taking root afterwards in 30 days;
(5) acclimatization and transplants: the bottle seedling possessing transplanting condition that root media obtains is carried out hardening, bottle seedling to move under normal temperature after 4 ~ 5 days, open bottle cap 2 ~ 3 days, then the medium being attached to shoot root and fastening is washed away, transplant to filling by peat soil: perlite: cultivate in the container bag of the transplanting culture matrix of vermiculite=1:1:1, each container bag is transplanted 1 strain and to be taken root seedling, need by matrix compacting during transplanting, and carry out individual plant bagging, sooner or later spraying, guarantee growing environment is moist, transplant and cut off plastic sack two jiaos after 7 days, completely de-bag after 14 days, transplant and add up survival rate after 30 days.
2. a kind of daybreak Ce bamboo tissue culture rapid breeding method according to claim 1, it is characterized in that the inducing culture described in step (2) is: MS+3.0 ~ 8.0mg/L6-BA+0.1 ~ 1.0mg/L ZT+1.0 ~ 3.0g/L AC+ 15 ~ 30g/L sucrose+3.5 ~ 6.0g/L agar, pH is 5.4 ~ 5.8.
3. a kind of daybreak Ce bamboo tissue culture rapid breeding method according to claim 1, it is characterized in that the proliferated culture medium described in step (3) is: MS+0.001 ~ 0.01mg/L TDZ+1.0 ~ 3.0mg/L ZT+1.0 ~ 4.0mg/L 6-BA+15 ~ 30g/L sucrose+3.5 ~ 6.0g/L agar, pH is 5.4 ~ 5.8.
4. a kind of daybreak Ce bamboo tissue culture rapid breeding method according to claim 1, it is characterized in that the root media described in step (4) is: MS+0.1 ~ 1.0mg/L IBA+1.0 ~ 3.0mg/L IAA+15 ~ 30g/L sucrose+3.5 ~ 6.0g/L agar, pH is 5.4 ~ 5.8.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108174786A (en) * | 2018-01-18 | 2018-06-19 | 广西正匠农业科技有限公司 | A kind of yushania method for tissue culture |
| CN109329029A (en) * | 2018-12-06 | 2019-02-15 | 四川农业大学 | It is suitble to the ground of laboratory scientific research by bamboo water planting breeding method and its breeding apparatus |
| CN114009342A (en) * | 2021-12-17 | 2022-02-08 | 宜宾林竹产业研究院 | Tissue culture and rapid propagation medium and tissue culture and rapid propagation method for viola tinctoria bamboo |
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| CN103314848A (en) * | 2013-05-29 | 2013-09-25 | 南京林业大学 | Rapidly breeding method for ampelocalamus luodianensis |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108174786A (en) * | 2018-01-18 | 2018-06-19 | 广西正匠农业科技有限公司 | A kind of yushania method for tissue culture |
| CN109329029A (en) * | 2018-12-06 | 2019-02-15 | 四川农业大学 | It is suitble to the ground of laboratory scientific research by bamboo water planting breeding method and its breeding apparatus |
| CN109329029B (en) * | 2018-12-06 | 2021-06-08 | 四川农业大学 | Ground cover bamboo water culture cultivation method and cultivation device suitable for laboratory research |
| CN114009342A (en) * | 2021-12-17 | 2022-02-08 | 宜宾林竹产业研究院 | Tissue culture and rapid propagation medium and tissue culture and rapid propagation method for viola tinctoria bamboo |
| CN114009342B (en) * | 2021-12-17 | 2022-07-01 | 宜宾林竹产业研究院 | Tissue culture and rapid propagation medium and tissue culture and rapid propagation method for viola tinctoria bamboo |
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Application publication date: 20150617 |