CN102657091A - Taxus chinensis var. maire embryo in-vitro seedling forming technique and culture medium employed by same - Google Patents
Taxus chinensis var. maire embryo in-vitro seedling forming technique and culture medium employed by same Download PDFInfo
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- CN102657091A CN102657091A CN2012101565876A CN201210156587A CN102657091A CN 102657091 A CN102657091 A CN 102657091A CN 2012101565876 A CN2012101565876 A CN 2012101565876A CN 201210156587 A CN201210156587 A CN 201210156587A CN 102657091 A CN102657091 A CN 102657091A
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- yew
- maire
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Abstract
The invention discloses a Taxus chinensis var. maire embryo in-vitro seedling forming technique, which comprises the following steps: removing aril from Taxus chinensis var. maire seeds, disinfecting by using alcohol and sodium hypochlorite in sequence, removing hard testa, taking embryos out, cleaning the embryos in sterile water, inoculating on a specific test-tube culture medium, performing dark culture, culturing under a lamp into seedlings after the embryos are sprouted, putting test-tube plantlets in an environment at the temperature of 25DEG C, light intensity of 1,500lx, illumination of 12h/d and relative humidity (RH) of about 80 percent for 7 days to complete in-bottle seedling hardening, transplanting the test-tube plantlets to a specific medium, controlling the temperatures at day and night to 20-25DEG C and 15-18DEG C, the light intensity at noon to 25-40klx and the RH to 70-85 percent, completing out-bottle medium seedling hardening after 25 days, and transplanting the test-tube plantlets to the field. By the Taxus chinensis var. maire embryo in-vitro seedling forming technique, the Taxus chinensis var. maire embryos are sprouted to form seedlings in the same year, and a quick method is provided for quickly propagating Taxus chinensis var. maire seedlings and protecting resources.
Description
Technical field
Southern enqlish yew embryo of the present invention exsomatizes into the seedling technology and belongs to biological technical field, relates to a kind of cultured in vitro technology and used medium thereof of southern enqlish yew embryo.
Background technology
Southern enqlish yew (
Taxus chinensis var. Maire), be China few survivors seeds in the distinctive Tertiary Period.Under nature, owing to reasons such as unisexual flower, dioecism, cross pollinatioies, the seed amount of formation is less; And the seed dormancy phase is long, needs one summer of two winters could rudiment under the natural conditions, and germination rate and seedling percent are low; Add its poor growth (one tree is become a useful person needs 60 years), in population, a little less than growth and the competitive ability, cause its natural distributed fewer and feweri.In addition,, southern enqlish yew uses and medical value because having special material, the destruction of its going to wreck property of resource since the eighties in 20th century, and existing resource seldom is in Critical Condition, and the State Administration of Forestry classified them as first class of protection seeds in 1992.Therefore, it is significant for protection and rational exploitation and utilization southern enqlish yew resource to seek fast and effectively southern enqlish yew sapling multiplication method.
Summary of the invention
The objective of the invention is provides a kind of southern enqlish yew embryo to exsomatize into the seedling technology in order to solve above-mentioned technical problem.
Technical scheme of the present invention
A kind of southern enqlish yew embryo exsomatizes into the seedling technology; Be about to embryo and directly from the southern enqlish yew seed, take out, in the stripped aseptic culture of the enterprising pedestrian worker of medium, the dormancy of breaking embryo; Impel it sprouting into seedling then, for the implant mass of southern enqlish yew provides approach efficiently.Be specifically related to cultivation, refining seedling and the field planting of the peeling off of embryo, embryo.
A kind of southern enqlish yew embryo exsomatizes into the seedling technology, specifically comprises the steps:
(1), southern enqlish yew seed embryo peels off
The southern enqlish yew seed is removed arillus, with alcohol, clorox sterilization, remove hard exosper successively, take out embryo, and embryo is rinsed well in sterile water;
(2), the cultivation of southern enqlish yew embryo
The embryo of step (1) gained is inoculated on the medium, under 20-22 ℃ of condition, secretly cultivates earlier, move to about 1500 1x of intensity of illumination after the sprouting, condition is cultured to root and grows under the light of illumination 12h/d, promptly gets the southern enqlish yew test-tube plantlet;
Used test tube culture medium prescription is by every liter of calculating, by 400mg NH
4N0
3, 556mg Ca (NO
3)
2.4H
2O, 990 mg K
2SO
4, 96 mg CaCl
2.2H
2O, 170 mg KH
2PO
4, 0.25 mg Na
2MoO
4.2H
2O, 370 mg MgSO
4.7H
2O, 22.4mg MnSO
4.H
2O, 8.6mg ZnSO
4.7H
2O, 0.25mg CuSO
4.5H
2O, 27.8 mg FeSO
4.7H
2O, 37.3 mg Na
2The water of-EDTA, 100 mg inositols, 1.0mg vitamin B1,0.5 mg nicotinic acid, 0.5 mg vitamin B6,2.0 mg glycine, 0.5mg gibberellin (GA), 0.5mg heteroauxin (IAA), 0.5mg 6-benzyl purine (6-BA), 20 g sucrose, 6g agar and 1g active carbon and surplus is formed;
(3), in-bottle seeding
The southern enqlish yew test-tube plantlet of step (2) gained is placed the environment of 25 ℃ of temperature, light intensity 1500lx, illumination 12h/d, RH about 80%, open bottle cap, make it the 7d that conforms, can accomplish in-bottle seeding;
(4), external substrate seeding
The test-tube plantlet of step (3) being accomplished in-bottle seeding takes out; Plant in matrix; Described matrix formulations is pressed mass ratio and is calculated, and promptly careless carbon: vermiculite: mountain mud is 2:2:6, and seeding room's day temperature is 20-25 ℃; Night be 15-18 ℃, noon light intensity 25-40klx, RH 70%-85%, can accomplish external substrate seeding through 25d;
(5), transplant the land for growing field crops
The plant of step (4) completion external substrate seeding is planted big Tanaka to get final product.
Beneficial effect of the present invention
A kind of southern enqlish yew embryo of the present invention exsomatizes into the seedling technology; Owing to adopted the technical scheme of embryo being separated and carrying out cultured in vitro from seed; Therefore can avoid germination inhibitor in the endosperm to the inhibitory action of seed germination, make embryo sprout into seedling then.Promptly solved and needed the restriction that to sprout naturally through one summer of two winters behind the southern enqlish yew seed collection, for the quick breeding of Chinese yew seedling provides method efficiently.
Embodiment
Through specific embodiment the present invention is further set forth below, but do not limit the present invention.
Southern enqlish yew seed in the embodiments of the invention picks up from first forest farm, mausoleum, Jingde, Anhui
Embodiment 1
A kind of southern enqlish yew embryo exsomatizes into the seedling technology, specifically comprises the steps:
(1), the sterilization of southern enqlish yew seed
The seed of gathering is then gone arillus, use distilled water immersion 24h, with 70% alcohol disinfecting 2min, 5% clorox sterilization 30min, aseptic water washing is totally subsequent use successively;
(2), the southern enqlish yew embryo is peeled off
Seed after the sterilization is placed on the aseptic filter paper, removed hard exosper, remove endosperm, and embryo is rinsed well in sterile water at microscopically;
(3), the cultivation of southern enqlish yew embryo
The embryo of step (2) gained is inoculated on the test tube medium, at first under 20-22 ℃ of condition, secretly cultivates, move to about 1500 1x of intensity of illumination after the sprouting, be cultured to root under the light of illumination 12h/d and grow, promptly get the southern enqlish yew test-tube plantlet;
Used test tube culture medium prescription is by every liter of calculating, by 0.5mgGA, 0.5mg IAA, 0.5mg6-BA, 400mg NH
4N0
3, 556mg Ca (NO
3)
2.4H
2O, 990 mg K
2SO
4, 96 mg CaCl
2.2H
2O, 170 mg KH
2PO
4, 0.25 mg Na
2MoO
4.2H
2O, 370 mg MgSO
4.7H
2O, 22.4mg MnSO
4.H
2O, 8.6mg ZnSO
4.7H
2O, 0.25mg CuSO
4.5H
2O, 27.8 mg FeSO
4.7H
2O, 37.3 mg Na
2The water of-EDTA, 100 mg inositols, 1.0mg vitamin B1,0.5 mg nicotinic acid, 0.5 mg vitamin B6,2.0 mg glycine, 20 g sucrose, 6g agar, 1g active carbon and surplus is formed, and transfers to pH5.6 with 1M NaOH or 1M HCI;
(4), in-bottle seeding
The southern enqlish yew test-tube plantlet of step (3) gained is placed the environment of 25 ℃ of temperature, light intensity 1500lx, illumination 12h/d, RH about 80%, open bottle cap, make it the 7d that conforms, can accomplish;
(5), external substrate seeding
The test-tube plantlet of step (4) being accomplished in-bottle seeding takes out, and plants in matrix, and day temperature is 20-25 ℃ in the matrix seeding room, night be 15-18 ℃, noon light intensity 25-40klx, RH 70 %-85 %, can accomplish through 25d;
Said matrix formulations is pressed mass ratio and is calculated, and promptly careless carbon: vermiculite: mountain mud is 2:2:6;
(6), transplant in the land for growing field crops
The plant of accomplishing external substrate seeding is planted big Tanaka, and final survival rate is 95%.
In sum; A kind of southern enqlish yew embryo of the present invention exsomatizes into the seedling technology and after the southern enqlish yew embryo is taken out, carry out culture in vitro from seed; Broken the sprouting restriction of endosperm to embryo, embryo can be sprouted at seed collection then, and is grown to serve as healthy seedling; After transplanted in refining seedling, land for growing field crops, survival rate reached 95%.The present invention is simple to operate, and cost is low, can be used as the method for the quick breeding of Chinese yew seedling.
Foregoing is merely the basic explanation of the present invention under conceiving, and according to any equivalent transformation that technical scheme of the present invention is done, all should belong to protection scope of the present invention.
Claims (4)
1. a southern enqlish yew embryo exsomatizes into the seedling technology, it is characterized in that specifically comprising the steps:
(1), southern enqlish yew seed embryo peels off
The southern enqlish yew seed is removed arillus, use earlier distilled water immersion, successively with alcohol, clorox sterilization, remove hard exosper then, take out embryo, and embryo is rinsed well in sterile water;
(2), the cultivation of southern enqlish yew embryo
The embryo of step (1) gained is inoculated on the test tube medium, under 20-22 ℃ of conditions, secretly cultivates earlier, move to about 1500 1x of intensity of illumination after the sprouting, be cultured to root under the condition of illumination 12h/d and grow, promptly obtain the southern enqlish yew test-tube plantlet;
Used test tube culture medium prescription is by every liter of calculating, by 400mg NH
4N0
3, 556mg Ca (NO
3)
2.4H
2O, 990 mg K
2SO
4, 96 mg CaCl
2.2H
2O, 170 mg KH
2PO
4, 0.25 mg Na
2MoO
4.2H
2O, 370 mg MgSO
4.7H
2O, 22.4mg MnSO
4.H
2O, 8.6mg ZnSO
4.7H
2O, 0.25mg CuSO
4.5H
2O, 27.8 mg FeSO
4.7H
2O, 37.3 mg Na
2The water of-EDTA, 100 mg inositols, 1.0mg vitamin B1,0.5 mg nicotinic acid, 0.5 mg vitamin B6,2.0 mg glycine, 0.5mg GA, 0.5mg IAA, 0.5mg 6-BA, 20 g sucrose, 6g agar and 1g active carbon and surplus is formed;
(3), in-bottle seeding
The southern enqlish yew test-tube plantlet of step (2) gained is placed the environment of 25 ℃ of temperature, light intensity 1500lx, illumination 12h/d, RH about 80%, open the test tube bottle cap, make it the 7d that conforms, can accomplish in-bottle seeding;
(4), external substrate seeding
The seedling of step (3) being accomplished the test tube of in-bottle seeding takes out, and plants and in the matrix of matrix seeding room, can accomplish external substrate seeding through 25d;
Described matrix formulations is pressed mass ratio and is calculated, and promptly careless carbon: vermiculite: mountain mud is 2:2:6;
Described matrix seeding room day temperature is 20-25 ℃, night be 15-18 ℃, noon light intensity 25-40klx, relative moisture be 70%-85%;
(5), transplant the land for growing field crops
The plant of step (4) completion external substrate seeding is planted big Tanaka to get final product.
2. a kind of southern enqlish yew embryo as claimed in claim 1 exsomatizes into the seedling technology; After it is characterized in that the Chinese yew seed described in the step (1) goes arillus; Use earlier distilled water immersion 24h, then successively with 70% alcohol disinfecting 2min, the 5% clorox 30min that sterilizes.
3. according to claim 1 or claim 2 a kind of southern enqlish yew embryo exsomatizes into the used Chinese yew embryo test tube medium of seedling technology, it is characterized in that used Chinese yew embryo test tube medium, by every liter of calculating, by 400mg NH
4N0
3, 556mg Ca (NO
3)
2.4H
2O, 990 mg K
2SO
4, 96 mg CaCl
2.2H
2O, 170 mg KH
2PO
4, 0.25 mg Na
2MoO
4.2H
2O, 370 mg MgSO
4.7H
2O, 22.4mg MnSO
4.H
2O, 8.6mg ZnSO
4.7H
2O, 0.25mg CuSO
4.5H
2O, 27.8 mg FeSO
4.7H
2O, 37.3 mg Na
2The water of-EDTA, 100 mg inositols, 1.0mg vitamin B1,0.5 mg nicotinic acid, 0.5 mg vitamin B6,2.0 mg glycine, 0.5mg GA, 0.5mg IAA, 0.5mg 6-BA, 20 g sucrose, 6g agar and 1g active carbon and surplus is formed.
4. Chinese yew embryo test tube medium as claimed in claim 3 is characterized in that the pH of Chinese yew embryo test tube medium is 5.6.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201210156587 CN102657091B (en) | 2012-05-21 | 2012-05-21 | Taxus chinensis var. maire embryo in-vitro seedling forming technique and culture medium employed by same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201210156587 CN102657091B (en) | 2012-05-21 | 2012-05-21 | Taxus chinensis var. maire embryo in-vitro seedling forming technique and culture medium employed by same |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102657091A true CN102657091A (en) | 2012-09-12 |
| CN102657091B CN102657091B (en) | 2013-10-16 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 201210156587 Expired - Fee Related CN102657091B (en) | 2012-05-21 | 2012-05-21 | Taxus chinensis var. maire embryo in-vitro seedling forming technique and culture medium employed by same |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103039365A (en) * | 2013-01-18 | 2013-04-17 | 通化师范学院 | Test tube one-step seedling method of taxus cuspidata |
| CN105638462A (en) * | 2015-12-30 | 2016-06-08 | 湖南农业大学 | Method for domesticating and transplanting taxus chinensis var mairei tissue culture seedling |
| CN106518350A (en) * | 2016-10-27 | 2017-03-22 | 六安市振清生态农业有限公司 | Nutritive organic matrix for taxus chinensis planting |
| CN110521602A (en) * | 2019-09-18 | 2019-12-03 | 江苏红豆杉健康科技股份有限公司 | A kind of Chinese yew Embryo Culture method for culturing seedlings |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH06296493A (en) * | 1993-04-14 | 1994-10-25 | Nippon Steel Corp | Method for producing taxol |
| JPH0751074A (en) * | 1993-08-13 | 1995-02-28 | Mitsui Petrochem Ind Ltd | Tissue culture of plant of genus taxus and production of taxane-type diterpene using the tissue culture method |
| JPH08298993A (en) * | 1995-05-02 | 1996-11-19 | Nippon Steel Corp | Production of taxane compound by tissue culture of taxus plant |
| CN1367254A (en) * | 2001-01-21 | 2002-09-04 | 沈阳泰泽植物细胞培养有限公司 | Method for producing taxol by means of cell culture |
| CN1454465A (en) * | 2003-04-04 | 2003-11-12 | 曹家骙 | Chinese yew sprout tissue culture medium and its sprout breeding method |
-
2012
- 2012-05-21 CN CN 201210156587 patent/CN102657091B/en not_active Expired - Fee Related
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH06296493A (en) * | 1993-04-14 | 1994-10-25 | Nippon Steel Corp | Method for producing taxol |
| JPH0751074A (en) * | 1993-08-13 | 1995-02-28 | Mitsui Petrochem Ind Ltd | Tissue culture of plant of genus taxus and production of taxane-type diterpene using the tissue culture method |
| JPH08298993A (en) * | 1995-05-02 | 1996-11-19 | Nippon Steel Corp | Production of taxane compound by tissue culture of taxus plant |
| CN1367254A (en) * | 2001-01-21 | 2002-09-04 | 沈阳泰泽植物细胞培养有限公司 | Method for producing taxol by means of cell culture |
| CN1454465A (en) * | 2003-04-04 | 2003-11-12 | 曹家骙 | Chinese yew sprout tissue culture medium and its sprout breeding method |
Non-Patent Citations (3)
| Title |
|---|
| 曾余力等: "南方红豆杉离体胚培养诱导不定芽研究", 《浙江林学院学报》 * |
| 汪智慧: "南方红豆杉的组织培养及其营养特性研究", 《中国优秀博硕士学位论文全文数据库 (硕士) 农业科技辑》 * |
| 臧新等: "激素对 2种红豆杉离体胚培养的影响激素对 2种红豆杉离体胚培养的影响", 《华中农业大学学报》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103039365A (en) * | 2013-01-18 | 2013-04-17 | 通化师范学院 | Test tube one-step seedling method of taxus cuspidata |
| CN103039365B (en) * | 2013-01-18 | 2014-04-16 | 通化师范学院 | Test tube one-step seedling method of taxus cuspidata |
| CN105638462A (en) * | 2015-12-30 | 2016-06-08 | 湖南农业大学 | Method for domesticating and transplanting taxus chinensis var mairei tissue culture seedling |
| CN105638462B (en) * | 2015-12-30 | 2018-05-04 | 湖南农业大学 | A kind of rooting culture method of southerm yew tissue-cultured seedling |
| CN106518350A (en) * | 2016-10-27 | 2017-03-22 | 六安市振清生态农业有限公司 | Nutritive organic matrix for taxus chinensis planting |
| CN110521602A (en) * | 2019-09-18 | 2019-12-03 | 江苏红豆杉健康科技股份有限公司 | A kind of Chinese yew Embryo Culture method for culturing seedlings |
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| CN102657091B (en) | 2013-10-16 |
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