CN110521602A - A kind of Chinese yew Embryo Culture method for culturing seedlings - Google Patents
A kind of Chinese yew Embryo Culture method for culturing seedlings Download PDFInfo
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- CN110521602A CN110521602A CN201910880118.0A CN201910880118A CN110521602A CN 110521602 A CN110521602 A CN 110521602A CN 201910880118 A CN201910880118 A CN 201910880118A CN 110521602 A CN110521602 A CN 110521602A
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- days
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- chinese yew
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/10—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material
- A01G24/12—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material containing soil minerals
- A01G24/15—Calcined rock, e.g. perlite, vermiculite or clay aggregates
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G31/00—Soilless cultivation, e.g. hydroponics
-
- 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
-
- 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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- Life Sciences & Earth Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Environmental Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Soil Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Inorganic Chemistry (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The present invention relates to a kind of method for propagation, and in particular to a kind of Chinese yew Embryo Culture method for culturing seedlings, comprising the following steps: explant processing, MS culture medium culture, Tube propagation, acclimatization and transplants.Of the invention separates from seed by embryo and carries out in vitro culture, avoids the influence that natural environment sprouts it；Cultivation stage uses different culture medium, and culture medium is added different material, improves germination rate and planting percent, provides efficiently method for Chinese yew breeding.
Description
Technical field
The present invention relates to a kind of method for propagation, and in particular to a kind of Chinese yew Embryo Culture method for culturing seedlings.
Background technique
Taxus is in shallow rooted plant, and main root is unobvious, lateral root is flourishing, is generally acknowledged endangered natural treasure in the world
Dilute plants for anticancer, since growth of taxol speed is slow under field conditions (factors), power of regeneration is poor, so since some time, generation
It is formed not yet within the scope of boundary and advises the Chinese yew raw material forestbase touched greatly.
There are two types of the common propagation methods of Chinese yew, seminal propagation and cutting propagation.Wherein seminal propagation need through
A pregermination procedure is crossed, gets wet sand storage Stratificated treatment seed naturally frequently with outdoor to improve percentage of seedgermination.However natural ring
Border is difficult artificially to control, such as does not rain that will lead to humidity lower for a long time, meet rainy weather again long-term rainfall cause humidity compared with
Greatly.And the cutting propagation of Chinese yew light, temperature, in terms of requirement it is relatively high, natural environment is unable to satisfy its breeding and wants
It asks, more human and material resources need to be put into, and survival rate is lower with respect to seminal propagation.
Summary of the invention
Present invention seek to address that the above problem, provides a kind of Chinese yew Embryo Culture method for culturing seedlings, avoids nature ring
The influence that it is bred in border substantially increases the germination rate and planting percent of Chinese yew.
Technical solution according to the invention, the Chinese yew Embryo Culture method for culturing seedlings, comprising the following steps:
(1) mature Chinese yew seed removes aril, impregnates 1-2 h in redistilled water after linear tap water flushing 5-10min,
Again successively with 70% alcohol disinfecting 1-2min, 0.1% mercuric chloride disinfection 20-30min, aseptic water washing is clean, on aseptic filter paper
It regard embryo removing as explant；
(2) explant obtained by step (1) is inoculated into MS culture medium, dark culture 15-20 days is to outer under conditions of 24-26 DEG C
Implant is sprouted；0.8-1.2g caseinhydrolysate, 0.8-1.2g yeast extract, 28-32g sucrose and 6-8g are added in every liter of MS culture medium
Agar；
(3) explant that will be sprouted obtained by step (2), moves under illumination condition and cultivates 20-25 days to the long 2cm of bud, cultivation temperature
It is 24-26 DEG C, daily illumination 10-12h, illuminance 2000lx；
(4) seedling obtained by step (3) is transferred in test tube and is individually cultivated 20-30 days, the WPM training of 1/4-1/2 is added in test tube
Support base；In the WPM culture medium of every liter of 1/4-1/2 add 0.1-0.5mg 6-BA, 0.05-0.1mg NAA, 4-6g activated carbon,
0.06-0.10g vitamin B1,28-32g sucrose and 6-8g agar；
(5) by into sterilization matrix, growth is transplanted to big after 25-40 days for transplanting after test tube plantlet hardening 4-7 days obtained by step (4)
Field.
Further, the sterilization matrix in the step (5) is vermiculite matrix.
The beneficial effects of the present invention are: embryo is separated to from seed and carried out in vitro culture, avoids natural environment pair
Its influence sprouted；Cultivation stage uses different culture medium, and culture medium is added different material, improves germination rate and planting percent,
Efficiently method is provided for Chinese yew breeding.
Specific embodiment
The present invention is further explained in the light of specific embodiments.
Embodiment one
A kind of Chinese yew Embryo Culture method for culturing seedlings, comprising the following steps:
(1) mature taxus chinensis in northeast seed removes aril, impregnates 1 h in redistilled water after rinsing 5min with linear tap water,
Again successively with 70% alcohol disinfecting 1min, 0.1% mercuric chloride disinfection 20min, aseptic water washing is clean, shells embryo on aseptic filter paper
From as explant；
(2) explant obtained by step (1) is inoculated into MS culture medium, dark culture 150 days is to explant under conditions of 24 DEG C
It sprouts；Lg caseinhydrolysate, 1g yeast extract, 30g sucrose and 7g agar are added in every liter of MS culture medium；
(3) explant that will be sprouted obtained by step (2), moves under illumination condition and cultivates 20 days to the long 2cm of bud, cultivation temperature is
24- DEG C, daily illumination 10h, illuminance 2000lx；
(4) seedling obtained by step (3) is transferred in test tube and is individually cultivated 200 days, 1/4 WPM culture medium is added in test tube；Often
It rises and adds 0.15mg 6-BA, 0 .05mg NAA, 5g activated carbon, 0.08g vitamin B1,30g sucrose in 1/4 WPM culture medium
With 7g agar；
(5) by into the vermiculite matrix of sterilizing, growth is transplanted to after 25 days for transplanting after test tube plantlet hardening 4 days obtained by step (4)
Crop field.
The germination rate of taxus chinensis in northeast seed is 99.5% in the present embodiment, planting percent 56.2%.
Embodiment two
A kind of Chinese yew Embryo Culture method for culturing seedlings, comprising the following steps:
(1) mature taxus chinensis in northeast seed removes aril, impregnates 2 h in redistilled water after rinsing 10min with linear tap water,
Again successively with 70% alcohol disinfecting 2min, 0.1% mercuric chloride disinfection 30min, aseptic water washing is clean, shells embryo on aseptic filter paper
From as explant；
(2) explant obtained by step (1) is inoculated into MS culture medium, dark culture 20 days to explant is sprouted under conditions of 26 DEG C
Hair；0.8g caseinhydrolysate, 0.8g yeast extract, 28g sucrose and 6g agar are added in every liter of MS culture medium；
(3) explant that will be sprouted obtained by step (2), moves under illumination condition and cultivates 25 days to the long 2cm of bud, cultivation temperature 26
DEG C, daily illumination 12h, illuminance 2000lx；
(4) seedling obtained by step (3) is transferred in test tube and is individually cultivated 20-30 days, 1/2 WPM culture medium is added in test tube；
0 .5mg 6-BA, 0 .1mg NAA, 4g activated carbon, 0.06g vitamin B1,28g sucrose are added in every liter 1/2 of WPM culture medium
With 6g agar；
(5) by into the vermiculite matrix of sterilizing, growth is transplanted to after 40 days for transplanting after test tube plantlet hardening 7 days obtained by step (4)
Crop field.
The germination rate of taxus chinensis in northeast seed is 99.3% in the present embodiment, planting percent 64.9%.
Embodiment three
A kind of Chinese yew Embryo Culture method for culturing seedlings, comprising the following steps:
(1) mature taxusyunnanensis seed removes aril, impregnates 1.5 in redistilled water after rinsing 70min with linear tap water
H, then successively with 70% alcohol disinfecting 1.5min, 0.1% mercuric chloride disinfection 25min, aseptic water washing is clean, will on aseptic filter paper
Embryo removing is used as explant；
(2) explant obtained by step (1) is inoculated into MS culture medium, dark culture 17 days to explant is sprouted under conditions of 25 DEG C
Hair；1.2g caseinhydrolysate, 1.2g yeast extract, 32g sucrose and 8g agar are added in every liter of MS culture medium；
(3) explant that will be sprouted obtained by step (2), moves under illumination condition and cultivates 23 days to the long 2cm of bud, cultivation temperature 25
DEG C, daily illumination 11h, illuminance 2000lx；
(4) seedling obtained by step (3) is transferred in test tube and is individually cultivated 23 days, 1/4 WPM culture medium is added in test tube；Often
Rise 1/4 WPM culture medium in add 0.3mg 6-BA, 0.08mg NAA, 6g activated carbon, 0.10g vitamin B1,32g sucrose and
8g agar；
(5) by into the vermiculite matrix of sterilizing, growth is transplanted to after 32 days for transplanting after test tube plantlet hardening 6 days obtained by step (4)
Crop field.
The germination rate of taxusyunnanensis seed is 99.7% in the present embodiment, planting percent 67.3%.
Claims (2)
1. a kind of Chinese yew Embryo Culture method for culturing seedlings, which comprises the following steps:
(1) mature Chinese yew seed removes aril, impregnates 1-2 h in redistilled water after linear tap water flushing 5-10min,
Again successively with 70% alcohol disinfecting 1-2min, 0.1% mercuric chloride disinfection 20-30min, aseptic water washing is clean, on aseptic filter paper
It regard embryo removing as explant；
(2) explant obtained by step (1) is inoculated into MS culture medium, dark culture 15-20 days is to outer under conditions of 24-26 DEG C
Implant is sprouted；0.8-1.2g caseinhydrolysate, 0.8-1.2g yeast extract, 28-32g sucrose and 6-8g are added in every liter of MS culture medium
Agar；
(3) explant that will be sprouted obtained by step (2), moves under illumination condition and cultivates 20-25 days to the long 2cm of bud, cultivation temperature
It is 24-26 DEG C, daily illumination 10-12h, illuminance 2000lx；
(4) seedling obtained by step (3) is transferred in test tube and is individually cultivated 20-30 days, the WPM training of 1/4-1/2 is added in test tube
Support base；In the WPM culture medium of every liter of 1/4-1/2 add 0.1-0.5mg 6-BA, 0.05-0.1mg NAA, 4-6g activated carbon,
0.06-0.10g vitamin B1,28-32g sucrose and 6-8g agar；
(5) by into sterilization matrix, growth is transplanted to big after 25-40 days for transplanting after test tube plantlet hardening 4-7 days obtained by step (4)
Field.
2. if the Chinese yew Embryo Culture method for culturing seedlings as described in right 1, which is characterized in that the sterilizing in the step (5)
Matrix is vermiculite matrix.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110972952A (en) * | 2019-12-26 | 2020-04-10 | 上海帝巨智能科技有限公司 | Tissue rapid propagation culture method of Chinese yew |
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| CN102657091A (en) * | 2012-05-21 | 2012-09-12 | 上海应用技术学院 | Taxus chinensis var. maire embryo in-vitro seedling forming technique and culture medium employed by same |
| CN102726292A (en) * | 2012-06-19 | 2012-10-17 | 湖南农业大学 | Method for rapidly breaking dormancy of taxales chinensis var.mairei seeds |
| CN104620985A (en) * | 2015-02-03 | 2015-05-20 | 北华大学 | Tissue culture and quick propagation method of taxus cuspidata |
| CN108703073A (en) * | 2018-05-29 | 2018-10-26 | 江苏红豆杉健康科技股份有限公司 | Southerm yew tissue cultivating and seedling method |
-
2019
- 2019-09-18 CN CN201910880118.0A patent/CN110521602A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102657091A (en) * | 2012-05-21 | 2012-09-12 | 上海应用技术学院 | Taxus chinensis var. maire embryo in-vitro seedling forming technique and culture medium employed by same |
| CN102726292A (en) * | 2012-06-19 | 2012-10-17 | 湖南农业大学 | Method for rapidly breaking dormancy of taxales chinensis var.mairei seeds |
| CN104620985A (en) * | 2015-02-03 | 2015-05-20 | 北华大学 | Tissue culture and quick propagation method of taxus cuspidata |
| CN108703073A (en) * | 2018-05-29 | 2018-10-26 | 江苏红豆杉健康科技股份有限公司 | Southerm yew tissue cultivating and seedling method |
Non-Patent Citations (5)
| Title |
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| JIN KANG等: "Plants as highly diverse sources of construction wood, handicrafts and fibre in the Heihe valley (Qinling Mountains, Shaanxi, China): the importance of minor forest products", 《JOURNAL OF ETHNOBIOLOGY AND ETHNOMEDICINE》 * |
| 李志良: "中国红豆杉和短叶红豆杉的胚胎培养", 《植物资源与环境学报》 * |
| 田启建等: "《湘西主要特色药用植物栽培与利用》", 31 May 2015, 西南交通大学出版社 * |
| 臧新等: "2种红豆杉的离体胚培养", 《郑州大学学报（理学版）》 * |
| 马均: "曼地亚红豆杉（Taxus Media cv.Hicksii）快繁技术体系研究", 《中国优秀硕士学位论文全文数据库农业科技辑》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110972952A (en) * | 2019-12-26 | 2020-04-10 | 上海帝巨智能科技有限公司 | Tissue rapid propagation culture method of Chinese yew |
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Application publication date: 20191203 |