CN103477981B - Acer palmatum Green Dragon tissue culture propagation technique - Google Patents
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- 241000931515 Acer palmatum Species 0.000 title claims abstract description 30
- 238000000034 method Methods 0.000 title claims abstract description 15
- 230000001954 sterilising effect Effects 0.000 claims abstract description 18
- 230000006698 induction Effects 0.000 claims abstract description 13
- 239000002609 medium Substances 0.000 claims abstract description 11
- 239000012879 subculture medium Substances 0.000 claims abstract description 11
- 238000005520 cutting process Methods 0.000 claims abstract description 7
- 230000035755 proliferation Effects 0.000 claims abstract description 7
- 238000009395 breeding Methods 0.000 claims abstract description 6
- 230000001488 breeding effect Effects 0.000 claims abstract description 6
- 201000010099 disease Diseases 0.000 claims abstract description 6
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims abstract description 6
- 238000012258 culturing Methods 0.000 claims description 15
- 238000005286 illumination Methods 0.000 claims description 15
- 239000000463 material Substances 0.000 claims description 15
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 5
- 229960002523 mercuric chloride Drugs 0.000 claims description 5
- LWJROJCJINYWOX-UHFFFAOYSA-L mercury dichloride Chemical compound Cl[Hg]Cl LWJROJCJINYWOX-UHFFFAOYSA-L 0.000 claims description 5
- 239000008223 sterile water Substances 0.000 claims description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 5
- 238000011081 inoculation Methods 0.000 claims 1
- 238000004659 sterilization and disinfection Methods 0.000 claims 1
- 241000700605 Viruses Species 0.000 abstract description 2
- 238000009825 accumulation Methods 0.000 abstract description 2
- 238000000338 in vitro Methods 0.000 abstract description 2
- 238000004519 manufacturing process Methods 0.000 abstract description 2
- 230000009286 beneficial effect Effects 0.000 abstract 1
- 241000208140 Acer Species 0.000 description 10
- 241001143500 Aceraceae Species 0.000 description 5
- 238000010586 diagram Methods 0.000 description 4
- 241000196324 Embryophyta Species 0.000 description 3
- 241000219227 Acer japonicum Species 0.000 description 2
- 244000046151 Acer negundo Species 0.000 description 2
- 235000004422 Acer negundo Nutrition 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- UUTKICFRNVKFRG-WDSKDSINSA-N (4R)-3-[oxo-[(2S)-5-oxo-2-pyrrolidinyl]methyl]-4-thiazolidinecarboxylic acid Chemical compound OC(=O)[C@@H]1CSCN1C(=O)[C@H]1NC(=O)CC1 UUTKICFRNVKFRG-WDSKDSINSA-N 0.000 description 1
- 235000015988 Acer ginnala Nutrition 0.000 description 1
- 240000004731 Acer pseudoplatanus Species 0.000 description 1
- 235000002754 Acer pseudoplatanus Nutrition 0.000 description 1
- 241000317410 Arisaema dracontium Species 0.000 description 1
- 235000002239 Dracunculus vulgaris Nutrition 0.000 description 1
- NNBFNNNWANBMTI-UHFFFAOYSA-M brilliant green Chemical group OS([O-])(=O)=O.C1=CC(N(CC)CC)=CC=C1C(C=1C=CC=CC=1)=C1C=CC(=[N+](CC)CC)C=C1 NNBFNNNWANBMTI-UHFFFAOYSA-M 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000035613 defoliation Effects 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 238000004161 plant tissue culture Methods 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000001172 regenerating effect Effects 0.000 description 1
- 241000894007 species Species 0.000 description 1
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Abstract
The invention discloses Acer palmatum Green Dragon tissue culture propagation technique, it comprises the following steps: S1, gather disease-free Acer palmatum Green Dragon current-year branch, removes unnecessary blade, is cut into stem with bud, sterilizing; S2, stem with bud after sterilizing is inoculated on medium grows, induction axillary bud sprouting; S3, shoot proliferation are cultivated, and are cut by the axillalry bud sprouted, are seeded on subculture medium, and induced bundle is sprouted generation; S4, culture of rootage, the indefinite bud on cutting Multiple Buds, is inoculated on root media, and induction root produces.The invention has the beneficial effects as follows: the tissue culture propagation system establishing the Green Dragon, achieve the breakthrough of Green Dragon breeding system; Low production cost, the plantlet in vitro obtained heredity shape is consistent, and reproduction coefficient is high, and the breeding cycle is short, is the numerous soon effective way of the Acer palmatum Green Dragon, and avoids virus accumulation.
Description
Technical field
The present invention relates to field of plant tissue culture technique, particularly Acer palmatum Green Dragon tissue culture propagation technique.
Background technology
Aceraceae (Aceraceae) Acer L (Acer) is general term maple usually, is accustomed to also known as " maple " or " maple ".Whole world maple has kind more than 200, is distributed in Asia, Europe, North America and Africa northern.China has more than 160 to plant, and account for more than 75% of whole world sum, all there is distribution all parts of the country, the main product Yangtze river basin and on the south each provinces and regions.Maple can be used as construction timber, paper making raw material, medicine extraction, iundustrial oil and nectariferous plant.Tree-like abundant, the attitude of some maple graceful, leaf abundant, leaf look changeable, is described as world-renowned ornamental foliage plant.Acer palmatum (Acer palmatum) has another name called Japanese maple, belongs to Aceraceae, Acer L defoliation small arbor or arbor.Tree performance grace is beautiful, leaf attractive in appearance, and leaf look changeable, is famous ornamental tree species.Countries in the world are introducing and planting already, mutation and modification a lot.Japan's Acer palmatum cultivates more than 450 reward leaf kinds at present.
The exploitation that China maple views and admires resource have very big-difference compared with abroad.Maple is as important foliage tree kind, and supply falls short of demand for cultivation nursery stock.Its breeding is main at present adopts cuttage and seed propagation method.Although Aceraceae plant has very large capacity in the growth in large field, carry out at present organize training breed of less types.
In sum, less to the Study on tissue culture of Aceraceae plant both at home and abroad, only there is a few kind to establish comparatively perfect axil proliferating tissues and cultivate regenerating system, as Acer pseudoplatanus, America Acer negundo, fine stern maple, Amur maple, Fructus Aceris davidii, Acer negundo. L.And report is rarely had for the Acer palmatum of cuttage root-taking difficulty and kind thereof.
The Acer palmatum Green Dragon (Acer palmatum ' Seiryu') is one of numerous Acer palmatum planting variation kind, its blade pinniform, and summer is emerald green, turns autumn red, flourishing, and strain shape is graceful, has much ornamental value.Be fancy breed comparatively general and precious on market today, but also report is had no precedent for the tissue culture propagation method of the Green Dragon.
Summary of the invention
The object of the invention is to the shortcoming overcoming prior art, the Acer palmatum Green Dragon tissue culture propagation technique that a kind of reproduction rate is high, reproduction speed is fast, easy and simple to handle is provided.
Object of the present invention is achieved through the following technical solutions: Acer palmatum Green Dragon tissue culture propagation technique, and it comprises the following steps:
S1, gather disease-free Acer palmatum Green Dragon current-year branch, remove unnecessary blade, be cut into stem with bud, stem with bud first uses alcohol sterilizing 30 ~ 60s, then uses mercuric chloride sterilizing 3 ~ 5min, then uses sterile water wash 4 ~ 6 times;
S2, stem with bud after sterilizing is inoculated on medium grows, described medium is MS+NAA 0.3mg/L, and postvaccinal culture materials is placed in intensity of illumination is 2000 ~ 3000LX, light application time is 16h/d, temperature is cultivate in the culturing room of 24 ~ 26 DEG C, cultivate 2 ~ 3 weeks, induction axillary bud sprouting;
S3, shoot proliferation are cultivated, the axillalry bud sprouted is cut, be seeded on subculture medium, subculture medium is MS+6-BA 1.5mg/L+NAA0.3mg/L, and postvaccinal culture materials is placed in intensity of illumination is 2000 ~ 3000LX, light application time is 16h/d, temperature is cultivate in the culturing room of 24 ~ 26 DEG C, cultivate 3 ~ 5 weeks, induced bundle is sprouted generation;
S4, culture of rootage, indefinite bud on cutting Multiple Buds, be inoculated on root media, root media is 1/2MS+NAA0.2mg/L, and postvaccinal culture materials is placed in intensity of illumination is 2000 ~ 3000LX, light application time is 16h/d, temperature is cultivate in the culturing room of 24 ~ 26 DEG C, cultivate 2 ~ 3 weeks, induction root produces.
The present invention has the following advantages:
The present invention, for gathering Green Dragon stem explants, Initial culture base induces axillalry bud to occur, then induced bundle is sprouted generation on proliferated culture medium, on cutting Multiple Buds, indefinite bud enters root media, root induction, establishes the tissue culture propagation system of the Green Dragon, achieves the breakthrough of Green Dragon breeding system.
Low production cost of the present invention, the plantlet in vitro obtained heredity shape is consistent, and reproduction coefficient is high, and the breeding cycle is short, is the numerous soon effective way of the Acer palmatum Green Dragon, and avoids virus accumulation.
Culture medium prescription of the present invention is simple, and cultivation flow process is simple and direct, easy and simple to handle, and culture efficiency is high, can be relatively easy to obtain Acer palmatum Green Dragon aseptic seedling, and hardening easily survives, time saving and energy saving.
Accompanying drawing explanation
Fig. 1 is the schematic diagram of Acer palmatum Green Dragon Initial culture of the present invention
Fig. 2 is the schematic diagram of Acer palmatum Green Dragon squamous subculture of the present invention
Fig. 3 is the schematic diagram of Acer palmatum Green Dragon culture of rootage of the present invention
Fig. 4 is the schematic diagram of Acer palmatum Green Dragon root system of the present invention.
Embodiment
Below in conjunction with drawings and Examples, the present invention will be further described, and protection scope of the present invention is not limited to the following stated:
embodiment 1:
Acer palmatum Green Dragon tissue culture propagation technique, it comprises the following steps:
S1, gather disease-free Acer palmatum Green Dragon current-year branch at fine day noon or afternoon, remove unnecessary blade, be cut into stem with bud, stem with bud first uses alcohol sterilizing 30s, then uses mercuric chloride sterilizing 5min, then uses sterile water wash 4 ~ 6 times;
S2, stem with bud after sterilizing is inoculated on medium grows, described medium is MS+NAA 0.3mg/L, and postvaccinal culture materials is placed in intensity of illumination is 2000 ~ 3000LX, light application time is 16h/d, temperature is cultivate in the culturing room of 24 ~ 26 DEG C, cultivate 15 days, induction axillary bud sprouting, as shown in Figure 1;
S3, shoot proliferation are cultivated, the axillalry bud sprouted is cut, be seeded on subculture medium, subculture medium is MS+6-BA 1.5mg/L+NAA0.3mg/L, and postvaccinal culture materials is placed in intensity of illumination is 2000 ~ 3000LX, light application time is 16h/d, temperature is cultivate in the culturing room of 24 ~ 26 DEG C, cultivate 30 days, induced bundle is sprouted generation, as shown in Figure 2;
S4, culture of rootage, indefinite bud on cutting Multiple Buds, be inoculated on root media, as shown in Figure 3, root media is 1/2MS+NAA0.2mg/L, and postvaccinal culture materials is placed in intensity of illumination is 2000 ~ 3000LX, light application time is 16h/d, temperature is cultivate in the culturing room of 24 ~ 26 DEG C, cultivates 15 days weeks, induction root produces, as shown in Figure 4.
embodiment 2:
Acer palmatum Green Dragon tissue culture propagation technique, it comprises the following steps:
S1, gather disease-free Acer palmatum Green Dragon current-year branch at fine day noon or afternoon, remove unnecessary blade, be cut into stem with bud, stem with bud first uses alcohol sterilizing 50s, then uses mercuric chloride sterilizing 4min, then uses sterile water wash 4 ~ 6 times;
S2, stem with bud after sterilizing is inoculated on medium grows, described medium is MS+NAA 0.3mg/L, and postvaccinal culture materials is placed in intensity of illumination is 2000 ~ 3000LX, light application time is 16h/d, temperature is cultivate in the culturing room of 24 ~ 26 DEG C, cultivate 2 weeks, induction axillary bud sprouting, as shown in Figure 1;
S3, shoot proliferation are cultivated, the axillalry bud sprouted is cut, be seeded on subculture medium, subculture medium is MS+6-BA 1.5mg/L+NAA0.3mg/L, and postvaccinal culture materials is placed in intensity of illumination is 2000 ~ 3000LX, light application time is 16h/d, temperature is cultivate in the culturing room of 24 ~ 26 DEG C, cultivate 3 weeks, induced bundle is sprouted generation, as shown in Figure 2;
S4, culture of rootage, indefinite bud on cutting Multiple Buds, be inoculated on root media, as shown in Figure 3, root media is 1/2MS+NAA0.2mg/L, and postvaccinal culture materials is placed in intensity of illumination is 2000 ~ 3000LX, light application time is 16h/d, temperature is cultivate in the culturing room of 24 ~ 26 DEG C, cultivate 2 weeks, induction root produces, as shown in Figure 4.
embodiment 3:
Acer palmatum Green Dragon tissue culture propagation technique, it comprises the following steps:
S1, gather disease-free Acer palmatum Green Dragon current-year branch at fine day noon or afternoon, remove unnecessary blade, be cut into stem with bud, stem with bud first uses alcohol sterilizing 60s, then uses mercuric chloride sterilizing 3min, then uses sterile water wash 4 ~ 6 times;
S2, stem with bud after sterilizing is inoculated on medium grows, described medium is MS+NAA 0.3mg/L, and postvaccinal culture materials is placed in intensity of illumination is 2000 ~ 3000LX, light application time is 16h/d, temperature is cultivate in the culturing room of 24 ~ 26 DEG C, cultivate 3 weeks, induction axillary bud sprouting, as shown in Figure 1;
S3, shoot proliferation are cultivated, the axillalry bud sprouted is cut, be seeded on subculture medium, subculture medium is MS+6-BA 1.5mg/L+NAA0.3mg/L, and postvaccinal culture materials is placed in intensity of illumination is 2000 ~ 3000LX, light application time is 16h/d, temperature is cultivate in the culturing room of 24 ~ 26 DEG C, cultivate 5 weeks, induced bundle is sprouted generation, as shown in Figure 2;
S4, culture of rootage, indefinite bud on cutting Multiple Buds, be inoculated on root media, as shown in Figure 3, root media is 1/2MS+NAA0.2mg/L, and postvaccinal culture materials is placed in intensity of illumination is 2000 ~ 3000LX, light application time is 16h/d, temperature is cultivate in the culturing room of 24 ~ 26 DEG C, cultivate 3 weeks, induction root produces, as shown in Figure 4.
Claims (1)
1. Acer palmatum Green Dragon tissue culture propagation method, it comprises: explant sterilization, inoculation and breeding, shoot proliferation cultivation and culture of rootage, and it is characterized in that, the concrete operations of described method are:
S1, gather disease-free Acer palmatum Green Dragon current-year branch, remove unnecessary blade, be cut into stem with bud, stem with bud first uses alcohol sterilizing 30 ~ 60s, then uses mercuric chloride sterilizing 3 ~ 5min, then uses sterile water wash 4 ~ 6 times;
S2, stem with bud after sterilizing is inoculated on medium grows, described medium is MS+NAA 0.3mg/L, and postvaccinal culture materials is placed in intensity of illumination is 2000 ~ 3000LX, light application time is 16h/d, temperature is cultivate in the culturing room of 24 ~ 26 DEG C, cultivate 2 ~ 3 weeks, induction axillary bud sprouting;
S3, shoot proliferation are cultivated, the axillalry bud sprouted is cut, be seeded on subculture medium, subculture medium is MS+6-BA 1.5mg/L+NAA0.3mg/L, and postvaccinal culture materials is placed in intensity of illumination is 2000 ~ 3000LX, light application time is 16h/d, temperature is cultivate in the culturing room of 24 ~ 26 DEG C, cultivate 3 ~ 5 weeks, induced bundle is sprouted generation;
S4, culture of rootage, indefinite bud on cutting Multiple Buds, be inoculated on root media, root media is 1/2MS+NAA0.2mg/L, and postvaccinal culture materials is placed in intensity of illumination is 2000 ~ 3000LX, light application time is 16h/d, temperature is cultivate in the culturing room of 24 ~ 26 DEG C, cultivate 2 ~ 3 weeks, induction root produces.
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| CN104285818A (en) * | 2014-10-30 | 2015-01-21 | 江苏省农业科学院 | Tissue culture rapid propagation method of acer palmatum |
| CN104663435B (en) * | 2015-02-09 | 2016-08-24 | 巴中七彩林业科技有限公司 | A kind of tissue culture and rapid propagation method of precious jade |
| CN104663436B (en) * | 2015-02-09 | 2016-06-15 | 巴中七彩林业科技有限公司 | A kind of tissue culture and rapid propagation method of red mirror |
| CN105638460B (en) * | 2015-12-30 | 2017-11-28 | 四川七彩林业开发有限公司 | A kind of tissue culture and rapid propagation method of Japanese maple butterfly |
| CN105409782B (en) * | 2015-12-30 | 2017-10-20 | 四川七彩林业开发有限公司 | A kind of yellow eight zhang tissue culture and rapid propagation method of mountain red autumnal leaves |
| CN109662032A (en) * | 2019-03-05 | 2019-04-23 | 延边大学 | A kind of culture medium and method of acer pseudo-sieboldianum tissue cultures |
| CN111011220A (en) * | 2020-01-10 | 2020-04-17 | 江苏农林职业技术学院 | Tissue culture rapid propagation method of beautiful maple |
| CN112544441A (en) * | 2020-11-10 | 2021-03-26 | 江苏省农业科学院 | Method for establishing tissue culture regeneration system of new acer palmatum variety |
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| CN101578963A (en) * | 2009-06-10 | 2009-11-18 | 重庆科技学院 | Tissue culture rapid propagation method for Japanese red maple |
| CN101846506A (en) * | 2010-05-07 | 2010-09-29 | 浙江大学 | Roll angle measurement method based on common path parallel beams |
| CN102668858A (en) * | 2012-06-05 | 2012-09-19 | 江苏省林业科学研究院 | Seedling cultivating method for cutting hard woods of acer palmatum |
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| USPP25302P3 (en) * | 2012-01-14 | 2015-02-24 | James David Cavett | Acer rubrum named ‘JSC Kingstwo’ |
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| CN101578963A (en) * | 2009-06-10 | 2009-11-18 | 重庆科技学院 | Tissue culture rapid propagation method for Japanese red maple |
| CN101846506A (en) * | 2010-05-07 | 2010-09-29 | 浙江大学 | Roll angle measurement method based on common path parallel beams |
| CN102668858A (en) * | 2012-06-05 | 2012-09-19 | 江苏省林业科学研究院 | Seedling cultivating method for cutting hard woods of acer palmatum |
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