CN102613083A - North American redwood tissue cultivation method - Google Patents
North American redwood tissue cultivation method Download PDFInfo
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- CN102613083A CN102613083A CN2012100992722A CN201210099272A CN102613083A CN 102613083 A CN102613083 A CN 102613083A CN 2012100992722 A CN2012100992722 A CN 2012100992722A CN 201210099272 A CN201210099272 A CN 201210099272A CN 102613083 A CN102613083 A CN 102613083A
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Abstract
The invention discloses a North American redwood tissue cultivation method, and belongs to a rapid vegetative propagation technology of agroforestrial seedlings. The North American redwood tissue cultivation method comprises the following steps of: disinfecting North American redwood twigs; and subjecting ordinary stem tips to induction, proliferation and rooting cultivation so as to obtain regenerated seedlings. Experiments show that: through induction and cultivation, differentiation rate of the stem tips can reach 100%, rooting rate of the stem tips is 75%, and transplanting survival rate of the stem tips is over 95%. The North American redwood tissue cultivation method has the beneficial effects that: as a resistant plant is taken as a parent plant, and the ordinary stem tips are taken as explant materials, genetic stability of excellent traits is ensured; sterilizing process is completed; varieties and concentration of suitable hormones are screened; the problems of long production cycle, high pollution rate, low rooting rate and low transplanting survival rate in the current studies are solved; and a North American redwood tissue cultivation system is successfully established.
Description
Technical field
The present invention relates to a kind of sequoia sempervirens tree method for tissue culture, is the asexual quick propagating technology of a kind of agricultural nursery stock.
Background technology
The sequoia sempervirens tree is that a kind of big footpath level also is important afforestation seeds with assortment.The breeding of present domestic sequoia sempervirens tree mainly is seminal propagation, and seed is mainly derived from U.S.'s import, and the kind amount is limited.Sequoia sempervirens tree cottage propagation rooting rate is low, and the side shoot cuttage seeding is because of the forfeiture apical dominance, and losing does not have value.Therefore through tissue culture technique, obtain a large amount of high-quality sequoia sempervirens tree seedling, promote significant the sequoia sempervirens tree.Domestic many scholars have carried out the research of sequoia sempervirens tree tissue culture now, but because rooting rate is low, breeding cycle length or explant are drawn materials and limited to its application.
Summary of the invention
In order to overcome the deficiency that exists in the present sequoia sempervirens tree tissue culture; The present invention provides a kind of sequoia sempervirens tree method for tissue culture, and not only the explant pollution rate is low for this method for tissue culture, and inductivity is high, appreciation rate is high, rooting rate is high; Breeding cycle is short, and transplanting survival rate is high.
The theoretical foundation of Plant Tissue Breeding is the plant cell Almightiness type; Be whole hereditary information that each cell of plant is all comprising these species; Thereby possess the hereditary potency that develops into whole plant, under optimum conditions, any one cell can develop into a new individuality.Select suitable explant, sterilization is inoculated in all kinds of cultivations under aseptic condition, sets up the aseptic culture system, and suitable temperature, illumination and humidity is provided, and cultivates into new individuality.
The technical solution adopted for the present invention to solve the technical problems is: a kind of sequoia sempervirens tree method for tissue culture; Utilize sequoia sempervirens tree spray; After sequoia sempervirens set spray and clean; Under aseptic condition, behind the alcohol disinfecting 30s of sequoia sempervirens tree spray after the cleaning with mass concentration 70%, mass concentration 0.1% HgCl
2Handle 15min, aseptic water washing 4-5 times is got the long common stem apex of 1cm and is done explant, the inoculation inducing culture, and pollution rate is lower than 5%, 26 ± 1 ℃ of cultivation temperature, humidity 70%, illumination 12h, intensity of illumination 2000Lx, incubation time are 40 days; The stem section of intercepting 1.5cm switching proliferated culture medium is cultivated 26 ± 1 ℃ of cultivation temperature, humidity 70%, illumination 12h, intensity of illumination 2000Lx then; Incubation time is 30 days; 1.5cm stem apex switching root media: 24 ± 1 ℃ of cultivation temperature, humidity 70%, illumination 12h, intensity of illumination 2000Lx; Incubation time is 50 days, and getting rooting rate is the tissue cultivating seedling more than 75%; Tissue cultivating seedling is cleaned the root medium, transplants in matrix, and matrix formulations is: turfy soil: perlite 3: 1, sterilized 1 hour down for 80 ℃; 24 ± 1 ℃ of acclimation temperatures, humidity 90%, illumination 12h, intensity of illumination 2000Lx; Reduce humidity after 10 days gradually, increase illumination, transplant the land for growing field crops after 20 days;
Described inducing culture is basal medium with MS, adds NAA 0.1mg/L, KT 0.5 mg/L, and sucrose 3%, agar 0.6%, pH 6.8;
Described proliferated culture medium is basal medium with MS, adds NAA 0.3mg/L, KT 0.5 mg/L, and BA 1.0 mg/L, sucrose 3%, agar 0.6%, pH 6.8;
Described root media is basal medium with 1/2MS, adds NAA 0.3mg/L, KT 0.5 mg/L, and BA 1.0 mg/L, sucrose 2%, agar 0.6%, pH 6.8.
The invention has the beneficial effects as follows: the present invention selects the resistance maternal plant to draw materials with the stem apex explant material, has guaranteed the genetic stability of merit; The perfect bacterium processing method that disappears; Filtered out suitable hormone kind and concentration simultaneously, it is long to have solved the production cycle that exists in the present research, and pollution rate is high, and rooting rate is low, and the problem that transplanting survival rate is low has successfully been set up the tissue culturing system of sequoia sempervirens tree.The stem apex differentiation rate reaches 100%, and rooting rate is 75%, and transplanting survival rate is more than 95%.
Embodiment
1, get sequoia sempervirens tree spray and wash 30min with 0.1% washing powder water logging after, it is subsequent use to wash 1h with running water.
2, under the aseptic condition, sequoia sempervirens tree spray with 70% alcohol disinfecting 30s after, 0.1% HgCl
2Handle 15min, aseptic water washing 4-5 times is got the long common stem apex of 1cm and is done explant, the inoculation inducing culture, and pollution rate is lower than 5%.Inducing culture is basal medium with MS, adds NAA0.1mg/L, KT0.5 mg/L, and sucrose 3%, agar 0.6%, pH 6.8,26 ± 1 ℃ of cultivation temperature, humidity 70%, illumination 12h, intensity of illumination 2000Lx.Incubation time is 40 days, the stem section switching proliferated culture medium of intercepting 1.5cm.The stem apex differentiation rate reaches 100%.
3, proliferated culture medium is basal medium with MS, adds NAA0.3mg/L, KT0.5 mg/L, and BA1.0 mg/L, sucrose 3%, agar 0.6%, pH 6.8,26 ± 1 ℃ of cultivation temperature, humidity 70%, illumination 12h, intensity of illumination 2000Lx.Incubation time is 30 days, gets the stem section switching proliferated culture medium of 1.5cm, the stem apex switching root media of 1.5cm.
4, root media is basal medium with 1/2MS, adds NAA0.3mg/L, KT0.5 mg/L, and BA1.0 mg/L, sucrose 2%, agar 0.6%, pH 6.8,24 ± 1 ℃ of cultivation temperature, humidity 70%, illumination 12h, intensity of illumination 2000Lx.Incubation time is 50 days, and rooting rate is more than 75%.
5, tissue cultivating seedling is cleaned the root medium, transplants in matrix the matrix formulations turfy soil: perlite 3: 1, sterilized 1 hour for 80 ℃; 24 ± 1 ℃ of acclimation temperatures, humidity 90%, illumination 12h, intensity of illumination 2000Lx; Reduce humidity after 10 days gradually, increase illumination, transplant the land for growing field crops after 20 days.Transplanting survival rate is more than 95%.
Claims (2)
1. sequoia sempervirens tree method for tissue culture is characterized in that: utilize sequoia sempervirens tree spray, and after sequoia sempervirens is set spray and cleans, under aseptic condition, behind the alcohol disinfecting 30s with mass concentration 70%, mass concentration 0.1% HgCl
2Handle 15min, aseptic water washing 4-5 times is got the long common stem apex of 1cm and is done explant, the inoculation inducing culture, and pollution rate is lower than 5%, 26 ± 1 ℃ of cultivation temperature, humidity 70%, illumination 12h, intensity of illumination 2000Lx, incubation time are 40 days; The stem section of intercepting 1.5cm switching proliferated culture medium is cultivated 26 ± 1 ℃ of cultivation temperature, humidity 70%, illumination 12h, intensity of illumination 2000Lx then; Incubation time is 30 days; 1.5cm stem apex switching root media: 24 ± 1 ℃ of cultivation temperature, humidity 70%, illumination 12h, intensity of illumination 2000Lx; Incubation time is 50 days, and getting rooting rate is the tissue cultivating seedling more than 75%; Tissue cultivating seedling is cleaned the root medium, transplants in matrix, and matrix formulations is: turfy soil: perlite 3: 1, sterilized 0.5 hour down for 80 ℃; 24 ± 1 ℃ of acclimation temperatures, humidity 90%, illumination 12h, intensity of illumination 2000Lx; Reduce humidity after 10 days gradually, increase illumination, transplant the land for growing field crops after 20 days;
Described inducing culture is basal medium with MS, adds NAA 0.1mg/L, KT 0.5 mg/L, and sucrose 3%, agar 0.6%, pH 6.8;
Described proliferated culture medium is basal medium with MS, adds NAA 0.3mg/L, KT 0.5 mg/L, and BA 1.0 mg/L, sucrose 3%, agar 0.6%, pH 6.8;
Described root media is basal medium with 1/2MS, adds NAA 0.3mg/L, KT 0.5 mg/L, and BA 1.0 mg/L, sucrose 2%, agar 0.6%, pH 6.8.
2. sequoia sempervirens according to claim 1 tree method for tissue culture is characterized in that: it is after sequoia sempervirens is set spray and washes 30min with 0.1% washing powder, to wash 1h with running water that described sequoia sempervirens tree spray cleans.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103798143A (en) * | 2014-02-25 | 2014-05-21 | 云南省农业科学院花卉研究所 | Out-bottle cutting and rooting method for sequoia sempervirens tissue culture seedling |
| CN104126492A (en) * | 2014-08-06 | 2014-11-05 | 江苏省中国科学院植物研究所 | In-vitro propagation method of taxodium hybrids 'zhongshansha136' |
| CN104304000A (en) * | 2014-08-29 | 2015-01-28 | 福建省林业科学研究院 | China fir clone tissue cultured seedling rooting induction method and rooting culture medium |
| CN105379627A (en) * | 2015-12-29 | 2016-03-09 | 江苏美尚生态景观股份有限公司 | Sequoia sempervirens in vitro rooting culture method |
| CN106359093A (en) * | 2016-08-31 | 2017-02-01 | 李军 | Establishing method for redwood tissue culture rapid propagation system |
| CN107996409A (en) * | 2018-01-25 | 2018-05-08 | 上海为绿景观建设有限公司 | A kind of stem apex sterilization method and sequoia sempervirens cultural method |
| CN108184669A (en) * | 2018-01-25 | 2018-06-22 | 上海为绿景观建设有限公司 | A kind of sequoia sempervirens culture medium and its cultural method |
| CN117617124A (en) * | 2024-01-23 | 2024-03-01 | 云南林业职业技术学院 | North American sequoia tissue culture rapid propagation method and application |
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103798143A (en) * | 2014-02-25 | 2014-05-21 | 云南省农业科学院花卉研究所 | Out-bottle cutting and rooting method for sequoia sempervirens tissue culture seedling |
| CN104126492A (en) * | 2014-08-06 | 2014-11-05 | 江苏省中国科学院植物研究所 | In-vitro propagation method of taxodium hybrids 'zhongshansha136' |
| CN104126492B (en) * | 2014-08-06 | 2016-04-13 | 江苏省中国科学院植物研究所 | The in-vitro propagation method of a kind of middle mountain China fir kind 136 |
| CN104304000A (en) * | 2014-08-29 | 2015-01-28 | 福建省林业科学研究院 | China fir clone tissue cultured seedling rooting induction method and rooting culture medium |
| CN104304000B (en) * | 2014-08-29 | 2016-08-31 | 福建省林业科学研究院 | A kind of Clones of Cunninghamia Lanceolata tissue cultured seedling root induction method and root media |
| CN105379627A (en) * | 2015-12-29 | 2016-03-09 | 江苏美尚生态景观股份有限公司 | Sequoia sempervirens in vitro rooting culture method |
| CN106359093A (en) * | 2016-08-31 | 2017-02-01 | 李军 | Establishing method for redwood tissue culture rapid propagation system |
| CN107996409A (en) * | 2018-01-25 | 2018-05-08 | 上海为绿景观建设有限公司 | A kind of stem apex sterilization method and sequoia sempervirens cultural method |
| CN108184669A (en) * | 2018-01-25 | 2018-06-22 | 上海为绿景观建设有限公司 | A kind of sequoia sempervirens culture medium and its cultural method |
| CN117617124A (en) * | 2024-01-23 | 2024-03-01 | 云南林业职业技术学院 | North American sequoia tissue culture rapid propagation method and application |
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Application publication date: 20120801 |