CN108174786A - A kind of yushania method for tissue culture - Google Patents
A kind of yushania method for tissue culture Download PDFInfo
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- CN108174786A CN108174786A CN201810048694.4A CN201810048694A CN108174786A CN 108174786 A CN108174786 A CN 108174786A CN 201810048694 A CN201810048694 A CN 201810048694A CN 108174786 A CN108174786 A CN 108174786A
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- culture
- yushania
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses a kind of yushania method for tissue culture, it carries out adventitious bud induction culture using being first inoculated in the first culture medium as explant into bamboo branch stem with bud, it is inoculated into the second culture medium after the completion of adventitious bud induction culture and carries out Multiplying culture, it is inoculated into third culture medium after the completion of Multiplying culture and carries out culture of rootage, the tissue-cultured seedling that will finally take root is placed at room temperature in daylight hardening 57 days, after being sterilized with 1000 times of carbendazim solutions in transplanting to fertile soil and perlite mixed-matrix, epiphragma or spraying and moisturizing after transplanting.Breeding culture is carried out to yushania using the method for the present invention, success rate is up to 78.6%, and tissue-cultured seedling is grown, and survival rate is up to 94.2%, and well developed root system, robust plant, growing way are strong.
Description
Technical field
The present invention relates to field of plant tissue culture technique, and in particular to a kind of yushania method for tissue culture.
Background technology
Yushania is also named Mount Taishan bamboo, is He undergraduate course bamboo Ya Ke le Sinobambusa plants.Bamboo plant height 8-15m, diameter 5-9cm, internode
Long 20-30cm, the interior tool of the several sections of sections of culm base, which is losed heart, takes root, and have canescence thin,tough silk hair up and down in sheaths of bamboo shoots ring.Stalk lower part starts branch, often
Joint number branch to racemosus fasciation, major branch is more slightly grown.Yushania originates in south of Yunnan, is born in low altitude area river bank or sparse woods more,
Guangxi, Guangdong, Hong Kong, Fujian, which have, draws cultivation.Leading bamboo pole is available for building, papermaking, can also make greenhouse bracket material, and bamboo shoots are delicious
Tasty, full of nutrition, bamboo clump is graceful, is usually used in Landscape.Yushania adaptability is stronger, in 120, world countries and regions
There is introducing and planting.Bamboo breeding is mainly to bury whip, bury stalk, bury section etc. based on traditional asexual reproduction method, but these methods
There are the problems such as breeding coefficient is low, labor intensity is big, seedling transport is inconvenient, can not meet needed for production.Tissue cultivating and seedling has
Have the advantages that breeding coefficient is big, quick, rejuvenation of preventing or cure a disease, and technically simple, it is of low cost, it can be received by industrial seedling rearing.
Invention content
In view of the above shortcomings of the prior art, the object of the present invention is to provide a kind of yushania method for tissue culture.
The technical solution adopted by the present invention is that：
A kind of yushania method for tissue culture, includes the following steps：
(1) explant is chosen, bamboo branch stem with bud is selected to, peels off blade and leaf sheath, cut twig, rushed with tap water
Then wash clean impregnates 10-20s again with aseptic water washing 2-3 times with 75% alcohol, then with aseptic water washing 2-3 times；
(2) explant is inoculated into the first culture medium and carries out adventitious bud induction culture, first culture medium is：Agar
10g/L, sucrose 30g/L, 6-BA1mg/L, IBA0.5mg/L, NAA0.05mg/L；
(3) it is inoculated into the second culture medium after the completion of adventitious bud induction culture and carries out Multiplying culture, second culture medium
For：Agar 10g/L, sucrose 30g/L, BA0.5mg/L, ZT0.2mg/L, NAA0.05mg/L；
(4) it is inoculated into third culture medium after the completion of Multiplying culture and carries out culture of rootage, the third culture medium is：1/
2MS、IBA 0.1mg/L、ABT0.1mg/L、NAA0.05mg/L；
(5) tissue-cultured seedling that will take root is placed at room temperature in daylight hardening 5-7 days, is moved after being sterilized with 1000 times of carbendazim solutions
It plants into fertile soil and perlite mixed-matrix, epiphragma or spraying and moisturizing after transplanting, the mass ratio of the fertile soil and perlite
It is 2:1-2.
Further, the condition of culture is：Day temperature is 25 ± 2 DEG C, 21 ± 1 DEG C of night temperatures, intensity of illumination
For 1500-2000lx, daily light application time is 12-14h.
Further, the adventitious bud induction culture time is 15-20d.
Further, the Multiplying culture time is 30-35d.
Further, the culture of rootage time is 12-15d.
It is an advantage of the invention that：Breeding culture is carried out to yushania using the method for the present invention, success rate is up to 78.6%,
Tissue-cultured seedling is grown, and survival rate is up to 94.2%, and well developed root system, robust plant, growing way are strong.
Specific embodiment
For a further understanding of the present invention, the preferred embodiment of the invention is described with reference to embodiment, still
It should be appreciated that these descriptions are only for the feature and advantage that further illustrate the present invention rather than to the claims in the present invention
Limitation.
Embodiment 1
A kind of yushania method for tissue culture, includes the following steps：
(1) explant is chosen, bamboo branch stem with bud is selected to, peels off blade and leaf sheath, cut twig, rushed with tap water
Then wash clean impregnates 10s again with aseptic water washing 2 times with 75% alcohol, then with aseptic water washing 2 times；
(2) it brings explant into laboratory to cultivate, primary condition is：Day temperature is 23 DEG C, 20 DEG C of night temperatures,
Intensity of illumination is 1500lx, and daily light application time is 12h；First, explant is inoculated into the first culture medium and carries out adventitious bud
Fiber differentiation 15d, first culture medium are：Agar 10g/L, sucrose 30g/L, 6-BA1mg/L, IBA0.5mg/L,
NAA0.05mg/L；
(3) it is inoculated into after the completion of adventitious bud induction culture in the second culture medium and carries out Multiplying culture 30d, second culture
Base is：Agar 10g/L, sucrose 30g/L, BA0.5mg/L, ZT0.2mg/L, NAA0.05mg/L；
(4) progress culture of rootage 12d, the third culture medium in third culture medium are inoculated into after the completion of Multiplying culture is：
1/2MS、IBA 0.1mg/L、ABT0.1mg/L、NAA0.05mg/L；
(5) tissue-cultured seedling that will take root is placed at room temperature in daylight hardening 5 days, is transplanted after being sterilized with 1000 times of carbendazim solutions
Into fertile soil and perlite mixed-matrix, epiphragma or spraying and moisturizing after transplanting, the mass ratio of the fertile soil and perlite is
2:1。
The present embodiment explant success rate is 75.4%, and tissue culture shoot survival percent is 92.9%.
Embodiment 2
A kind of yushania method for tissue culture, includes the following steps：
(1) explant is chosen, bamboo branch stem with bud is selected to, peels off blade and leaf sheath, cut twig, rushed with tap water
Then wash clean impregnates 15s again with aseptic water washing 3 times with 75% alcohol, then with aseptic water washing 3 times；
(2) it brings explant into laboratory to cultivate, primary condition is：Day temperature is 25 DEG C, 21 DEG C of night temperatures,
Intensity of illumination is 1800lx, and daily light application time is 13h；First, explant is inoculated into the first culture medium and carries out adventitious bud
Fiber differentiation 18d, first culture medium are：Agar 10g/L, sucrose 30g/L, 6-BA1mg/L, IBA0.5mg/L,
NAA0.05mg/L；
(3) it is inoculated into after the completion of adventitious bud induction culture in the second culture medium and carries out Multiplying culture 32d, second culture
Base is：Agar 10g/L, sucrose 30g/L, BA0.5mg/L, ZT0.2mg/L, NAA0.05mg/L；
(4) progress culture of rootage 13d, the third culture medium in third culture medium are inoculated into after the completion of Multiplying culture is：
1/2MS、IBA 0.1mg/L、ABT0.1mg/L、NAA0.05mg/L；
(5) tissue-cultured seedling that will take root is placed at room temperature in daylight hardening 6 days, is transplanted after being sterilized with 1000 times of carbendazim solutions
Into fertile soil and perlite mixed-matrix, epiphragma or spraying and moisturizing after transplanting, the mass ratio of the fertile soil and perlite is
2:1.5。
The present embodiment explant success rate is 78.6%, and tissue culture shoot survival percent is 94.2%.
Embodiment 3
A kind of yushania method for tissue culture, includes the following steps：
(1) explant is chosen, bamboo branch stem with bud is selected to, peels off blade and leaf sheath, cut twig, rushed with tap water
Then wash clean impregnates 20s again with aseptic water washing 3 times with 75% alcohol, then with aseptic water washing 3 times；
(2) it brings explant into laboratory to cultivate, primary condition is：Day temperature is 27 DEG C, 22 DEG C of night temperatures,
Intensity of illumination is 2000lx, and daily light application time is 14h；First, explant is inoculated into the first culture medium and carries out adventitious bud
Fiber differentiation 20d, first culture medium are：Agar 10g/L, sucrose 30g/L, 6-BA1mg/L, IBA0.5mg/L,
NAA0.05mg/L；
(3) it is inoculated into after the completion of adventitious bud induction culture in the second culture medium and carries out Multiplying culture 35d, second culture
Base is：Agar 10g/L, sucrose 30g/L, BA0.5mg/L, ZT0.2mg/L, NAA0.05mg/L；
(4) progress culture of rootage 15d, the third culture medium in third culture medium are inoculated into after the completion of Multiplying culture is：
1/2MS、IBA 0.1mg/L、ABT0.1mg/L、NAA0.05mg/L；
(5) tissue-cultured seedling that will take root is placed at room temperature in daylight hardening 7 days, is transplanted after being sterilized with 1000 times of carbendazim solutions
Into fertile soil and perlite mixed-matrix, epiphragma or spraying and moisturizing after transplanting, the mass ratio of the fertile soil and perlite is
2:2。
The present embodiment explant success rate is 75.3%, and tissue culture shoot survival percent is 93.5%.
Claims (5)
1. a kind of yushania method for tissue culture, it is characterised in that include the following steps：
(1) explant is chosen, bamboo branch stem with bud is selected to, peels off blade and leaf sheath, cut twig, is rinsed with tap water dry
Only again with aseptic water washing 2-3 times, then 10-20s is impregnated with 75% alcohol, then with aseptic water washing 2-3 times；
(2) explant is inoculated into the first culture medium and carries out adventitious bud induction culture, first culture medium is：Agar 10g/
L, sucrose 30g/L, 6-BA1mg/L, IBA0.5mg/L, NAA0.05mg/L；
(3) it is inoculated into the second culture medium after the completion of adventitious bud induction culture and carries out Multiplying culture, second culture medium is：Fine jade
Fat 10g/L, sucrose 30g/L, BA0.5mg/L, ZT0.2mg/L, NAA0.05mg/L；
(4) it is inoculated into third culture medium after the completion of Multiplying culture and carries out culture of rootage, second culture medium is：1/2MS、
IBA 0.1mg/L、ABT0.1mg/L、NAA0.05mg/L；
(5) tissue-cultured seedling that will take root is placed at room temperature in daylight hardening 5-7 days, transplanted after being sterilized with 1000 times of carbendazim solutions to
In fertile soil and perlite mixed-matrix, epiphragma or spraying and moisturizing after transplanting, the mass ratio of the fertile soil and perlite is 2:
1-2。
2. yushania method for tissue culture according to claim 1, it is characterised in that the condition of culture is：Daytime temperature
It is 25 ± 2 DEG C to spend, 21 ± 1 DEG C of night temperatures, intensity of illumination 1500-2000lx, and daily light application time is 12-14h.
3. yushania method for tissue culture according to claim 1, it is characterised in that the adventitious bud induction culture time is 15-
20d。
4. yushania method for tissue culture according to claim 1, it is characterised in that the Multiplying culture time is 30-35d.
5. yushania method for tissue culture according to claim 1, it is characterised in that the culture of rootage time is 12-15d.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201810048694.4A CN108174786A (en) | 2018-01-18 | 2018-01-18 | A kind of yushania method for tissue culture |
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| CN201810048694.4A CN108174786A (en) | 2018-01-18 | 2018-01-18 | A kind of yushania method for tissue culture |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP4292426A1 (en) * | 2022-06-15 | 2023-12-20 | Plant Select NV | Juvenile bamboo plant of the genus fargesia |
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|---|---|---|---|---|
| WO2000053727A1 (en) * | 1999-02-18 | 2000-09-14 | Johan Gielis | Micropropagation, synthetic seeds and germplasm storage of bamboos |
| CN101336613A (en) * | 2008-08-12 | 2009-01-07 | 浙江林学院 | Black bamboo tissue culture medium and tissue culture and rapid propagation method |
| WO2013016198A1 (en) * | 2011-07-22 | 2013-01-31 | Booshoot Llc | Compositions, methods, and systems for micropropagation of plants |
| CN102919124A (en) * | 2012-11-11 | 2013-02-13 | 云南禾起生物科技有限公司 | Rapid propagation method for dendrocalamus giganteus tissue culture seedling industrial production |
| CN103314848A (en) * | 2013-05-29 | 2013-09-25 | 南京林业大学 | Rapidly breeding method for ampelocalamus luodianensis |
| CN103907535A (en) * | 2014-04-16 | 2014-07-09 | 南京林业大学 | Method for obtaining large number of bambusa glaucophylla regeneration plants through tissue culture |
| CN104705186A (en) * | 2015-03-10 | 2015-06-17 | 朱海燕 | Tissue culture and quick propagation method of S. versicolor |
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2018
- 2018-01-18 CN CN201810048694.4A patent/CN108174786A/en not_active Withdrawn
Patent Citations (7)
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|---|---|---|---|---|
| WO2000053727A1 (en) * | 1999-02-18 | 2000-09-14 | Johan Gielis | Micropropagation, synthetic seeds and germplasm storage of bamboos |
| CN101336613A (en) * | 2008-08-12 | 2009-01-07 | 浙江林学院 | Black bamboo tissue culture medium and tissue culture and rapid propagation method |
| WO2013016198A1 (en) * | 2011-07-22 | 2013-01-31 | Booshoot Llc | Compositions, methods, and systems for micropropagation of plants |
| CN102919124A (en) * | 2012-11-11 | 2013-02-13 | 云南禾起生物科技有限公司 | Rapid propagation method for dendrocalamus giganteus tissue culture seedling industrial production |
| CN103314848A (en) * | 2013-05-29 | 2013-09-25 | 南京林业大学 | Rapidly breeding method for ampelocalamus luodianensis |
| CN103907535A (en) * | 2014-04-16 | 2014-07-09 | 南京林业大学 | Method for obtaining large number of bambusa glaucophylla regeneration plants through tissue culture |
| CN104705186A (en) * | 2015-03-10 | 2015-06-17 | 朱海燕 | Tissue culture and quick propagation method of S. versicolor |
Non-Patent Citations (5)
| Title |
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| G.R.ROUT等: "In vitro plant regeneration via callogenesis and organogenesis in Bambusa vulgaris", 《BIOLOGIA PLANTARUM》 * |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP4292426A1 (en) * | 2022-06-15 | 2023-12-20 | Plant Select NV | Juvenile bamboo plant of the genus fargesia |
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Application publication date: 20180619 |