CN104663450A - Tissue culture and rapid propagation method for Acer rubrum 'Brandywine' - Google Patents
Tissue culture and rapid propagation method for Acer rubrum 'Brandywine' Download PDFInfo
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- CN104663450A CN104663450A CN201510097698.8A CN201510097698A CN104663450A CN 104663450 A CN104663450 A CN 104663450A CN 201510097698 A CN201510097698 A CN 201510097698A CN 104663450 A CN104663450 A CN 104663450A
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Abstract
The invention discloses a tissue culture and rapid propagation method for Acer rubrum 'Brandywine'. The Acer rubrum 'Brandywine' is an Aceraceae Acer tall deciduous tree, has the characteristics of high adaptability, drought resistance, temperature resistance, growth fastness and the like and is a fine tree species for landscaping. At present, the Acer rubrum 'Brandywine' mainly propagates seedlings with a seed sowing propagation method, but the seed fruiting rate is lower, so that supply of a large number of seedlings of the Acer rubrum 'Brandywine' is limited. According to the method, stems with buds are used as explants, tissue-cultured regenerated plants of the Acer rubrum 'Brandywine' are successfully obtained through steps of induction, multiplication, rooting, acclimatization, transplantation and the like, a systematic tissue-cultured plant regeneration system for the Acer rubrum 'Brandywine' is established, and a reference is provided for mass factory culture of the seedlings of the Acer rubrum 'Brandywine'.
Description
Technical field
The present invention relates to the method for Plant Tissue Breeding in agricultural biotechnologies, specifically, relate to a kind of American red-maple tissue culture and rapid propagation method.
Background technology
American red-maple (Acer rubrum '
brandywine'), have another name called red maple, the red maple in North America, Canada Red maple, northern red maple, scarlet maple, scarlet maple, marsh maple etc., be the tall and big deciduous tree of Aceraceae Acer L, having the features such as strong adaptability, drought-enduring, heatproof, growing way are swift and violent, is the fine tree species of afforestation.Spring, young leaves was general red, mutually shone with the red flower of bunchiness; Summer, branches and leaves were lined: autumn, leaf was that azarin turns over yellow, to finally presenting cerise, heard long when continuing.Tree-like upright and on, tree crown is circular, grey bark.Annual various colors, spring blooms, monoecism, premium look, and fruit has wing, red.The transplanting survival rate of American red-maple is very high, and growth potential is strong, and have resistance, can adapt to the particular surroundings in street, cultivation management workload is little, and American red-maple damage by disease and insect is little, and after finding, process can not impact trees in time.American red-maple has many advantages of plane tree, very popular, and American red-maple gardening kind has become the outstanding Afforestation Landscape seeds of alternative fragrant camphor tree and Chinese parasol tree, is " king of street tree " of new generation.
Current American red-maple mainly carries out seedling breeding with planting seed modes of reproduction, but because of seed-setting rate lower, because which limit the supply of a large amount of seedling of American red-maple.Plant Tissue Breeding has that breeding is fast, cost less, growth coefficient high; the present invention take stem with bud as explant; through inducing, breed, to take root and the step such as acclimatization and transplants successfully obtains American red-maple Regenerated Plantlets; establishing the red mangrove tissue culture plants regenerating system of system, providing reference for realizing the nursery of American red-maple seedling scale factory.
Summary of the invention
The object of the present invention is to provide out a kind of American red-maple tissue culture and rapid propagation method, the present invention take stem with bud as explant, through inducing, breed, to take root and the step such as acclimatization and transplants successfully obtains American red-maple Regenerated Plantlets, establish the red mangrove tissue culture plants regenerating system of system, thus achieve object of the present invention.
A kind of American red-maple tissue culture and rapid propagation method of the present invention, comprises the following steps:
(1) explant sterilization: gather the stem Duan Yi Cheongju water of returning and to rinse after 10 ~ 25min and to brush away impurity above gently with hairbrush, with the 10 ~ 30s that sterilizes in 70% ~ 80% ethanolic solution in superclean bench, afterwards with aseptic washing 3 ~ 5 times, again with 0.1% ~ 0.3% mercuric chloride solution sterilization 10 ~ 25min, with for subsequent use after sucking moisture with aseptic filter paper after aseptic water washing 4 ~ 6 times.
(2) Fiber differentiation: the stem section that step (1) disinfects is cut into band 1 stipes and is about the long stem section of 1.5cm and is inoculated in inducing culture and carry out bud inducement cultivation.First full light culture 5 ~ 10 days under 25 ~ 28 DEG C of conditions after inoculation, be then placed in illumination every day 10 ~ 14 hours, intensity of illumination is 2000 ~ 3000lx, and being placed in cultivation temperature is cultivate under the condition of 25 ~ 28 DEG C until formation indefinite bud.
(3) Multiplying culture: step (2) is cultivated the bud seedling obtained, cut bud from base portion and proceed to proliferated culture medium and carry out squamous subculture, first full light culture 2 ~ 3 days under 25 ~ 28 DEG C of conditions after inoculation, then illumination every day is placed in 11 ~ 15 hours, intensity of illumination is 2000 ~ 3000lx, being placed in cultivation temperature is cultivate under the condition of 25 ~ 28 DEG C, and 30 days subcultures once.
(4) culture of rootage: when the bud seedling of step (2) or (3) is grown to 1.5 ~ 2.5cm height, cut into simple bud and be inoculated on root media and carry out culture of rootage, first full light culture 2 ~ 3 days under 25 ~ 28 DEG C of conditions after inoculation, then illumination every day is placed in 14 ~ 16 hours, intensity of illumination is 3000 ~ 5000lx, and cultivation temperature is be cultured under the condition of 25 ~ 28 DEG C to take root.
(5) acclimatization and transplants: the blake bottle bottle cap growing to the rooting tube plantlet of 3 ~ 4cm is opened and is placed in natural lighting lower refining seedling after 5 ~ 7 days, test-tube plantlet is taken out from blake bottle, wash root medium off, do not injure root as far as possible, plant in the matrix be mixed into by yellow sand and fiery carbon mud (1:1) in illumination every day 10 ~ 12 hours, intensity of illumination is 3000 ~ 5000lx, cultivation temperature is 25 ~ 28 DEG C, air humidity is cultivate 5 ~ 7 days in the incubator of 80% ~ 90%, and then transplant planting is in large Tanaka.
Inducing culture described in above-mentioned steps (2) is: WPM+1.0 ~ 5.0mg/L 6-BA+0.1 ~ 1.0mg/LIBA+15 ~ 30g/L sucrose+3.5 ~ 6.0g/L agar, pH is 5.4 ~ 5.8.
Proliferated culture medium described in above-mentioned steps (3) is: WPM+0.001 ~ 0.01mg/L TDZ+0.1 ~ 1.0mg/L NAA+15 ~ 30g/L sucrose+3.5 ~ 6.0g/L agar, pH is 5.4 ~ 5.8.
Root media described in above-mentioned steps (4) is: MS+1.0 ~ 5.0mg/L IBA+1.0 ~ 2.0mg/L GA
3+ 0.1 ~ 0.5mg/L 6-BA+15 ~ 30g/L sucrose+3.5 ~ 6.0g/L agar, pH is 5.4 ~ 5.8.
Advantage of the present invention is: American red-maple (Acer rubrum '
brandywine') be the tall and big deciduous tree of Aceraceae Acer L, having the features such as strong adaptability, drought-enduring, heatproof, growing way are swift and violent, is the fine tree species of afforestation.Current American red-maple mainly carries out seedling breeding with planting seed modes of reproduction, but because of seed-setting rate lower, because which limit the supply of a large amount of seedling of American red-maple.The present invention take stem with bud as explant; through inducing, breed, to take root and the step such as acclimatization and transplants successfully obtains American red-maple Regenerated Plantlets; establishing the American red-maple tissue culture plants regenerating system of system, providing reference for realizing the nursery of American red-maple seedling scale factory.
Embodiment
Following examples further illustrate of the present invention, is not limitation of the present invention.
embodiment 1
(1) explant sterilization: gather the stem Duan Yi Cheongju water of returning and to rinse after 10min and to brush away impurity above gently with hairbrush, with the 10s that sterilizes in 75% ethanolic solution in superclean bench, afterwards with aseptic washing 3 times, again with 0.1% mercuric chloride solution sterilization 10min, with for subsequent use after sucking moisture with aseptic filter paper after aseptic water washing 4 times.
(2) Fiber differentiation: the stem section that step (1) disinfects is cut into band 1 stipes and is about the long stem section of 1.5cm and is inoculated in inducing culture and carry out bud inducement cultivation.First full light culture 5 days under 25 DEG C of conditions after inoculation, be then placed in illumination every day 10 hours, intensity of illumination is 2000lx, being placed in cultivation temperature is cultivate until form indefinite bud under the condition of 25 DEG C, average pollution rate is for being only 4.5%, and average survival is 89.7%, and inductivity is 85.3%.Described inducing culture is: WPM+2.0mg/L 6-BA+0.1mg/LIBA+25g/L sucrose+4.5g/L agar, pH is 5.5.
(3) Multiplying culture: step (2) is cultivated the bud seedling obtained, cut bud from base portion and proceed to proliferated culture medium and carry out squamous subculture, first full light culture 2 days under 25 DEG C of conditions after inoculation, then illumination every day is placed in 11 hours, intensity of illumination is 2000lx, being placed in cultivation temperature is cultivate under the condition of 25 DEG C within 10 days, namely to can be observed the elongation growth of axillalry bud, growth coefficient be 3.8,30 days subcultures once.Described proliferated culture medium is: WPM+0.005mg/L TDZ+0.5mg/L NAA+15g/L sucrose+3.7g/L agar, pH is 5.5.
(4) culture of rootage: when the bud seedling of step (2) or (3) is grown to 1.5 ~ 2.5cm height, cut into simple bud and be inoculated on root media and carry out culture of rootage, first full light culture 2 days under 25 DEG C of conditions after inoculation, then illumination every day is placed in 14 hours, intensity of illumination is 3000lx, cultivation temperature is be cultured under the condition of 25 DEG C to take root, and rooting rate is 92.8%.Described root media is: MS+3.0mg/L IBA+1.2mg/L GA
3+ 0.1mg/L 6-BA+25g/L sucrose+4.0g/L agar, pH is 5.5.
(5) acclimatization and transplants: the blake bottle bottle cap growing to the rooting tube plantlet of 3 ~ 4cm is opened and is placed in natural lighting lower refining seedling after 5 days, test-tube plantlet is taken out from blake bottle, wash root medium off, do not injure root, plant in illumination every day 10 hours in the matrix be mixed into by yellow sand and fiery carbon mud (1:1), intensity of illumination is 3000lx as far as possible, cultivation temperature is 25 DEG C, air humidity is cultivate 5 days in the incubator of 80%, and then transplant planting is in large Tanaka, and transplanting survival rate is 91.7%.
Embodiment 2
(1) explant sterilization: gather the stem Duan Yi Cheongju water of returning and to rinse after 15min and to brush away impurity above gently with hairbrush, with the 20s that sterilizes in 75% ethanolic solution in superclean bench, afterwards with aseptic washing 5 times, again with 0.1% mercuric chloride solution sterilization 16min, with for subsequent use after sucking moisture with aseptic filter paper after aseptic water washing 6 times.
(2) Fiber differentiation: the stem section that step (1) disinfects is cut into band 1 stipes and is about the long stem section of 1.5cm and is inoculated in inducing culture and carry out bud inducement cultivation.First full light culture 7 days under 27 DEG C of conditions after inoculation, be then placed in illumination every day 14 hours, intensity of illumination is 3000lx, being placed in cultivation temperature is cultivate until form indefinite bud under the condition of 27 DEG C, average pollution rate is for being only 5.3%, and average survival is 90.4%, and inductivity is 86.7%.Described inducing culture is: WPM+4.0mg/L 6-BA+0.5mg/LIBA+28g/L sucrose+4.0g/L agar, pH is 5.7.
(3) Multiplying culture: step (2) is cultivated the bud seedling obtained, cut bud from base portion and proceed to proliferated culture medium and carry out squamous subculture, first full light culture 4 days under 27 DEG C of conditions after inoculation, then illumination every day is placed in 14 hours, intensity of illumination is 3000lx, being placed in cultivation temperature is cultivate under the condition of 27 DEG C within 10 days, namely to can be observed the elongation growth of axillalry bud, growth coefficient be 4.3,30 days subcultures once.Described proliferated culture medium is: WPM+0.01mg/L TDZ+1.0mg/L NAA+18g/L sucrose+4.0g/L agar, pH is 5.7.
(4) culture of rootage: when the bud seedling of step (2) or (3) is grown to 1.5 ~ 2.5cm height, cut into simple bud and be inoculated on root media and carry out culture of rootage, first full light culture 4 days under 27 DEG C of conditions after inoculation, then illumination every day is placed in 14 hours, intensity of illumination is 4000lx, cultivation temperature is be cultured under the condition of 27 DEG C to take root, and rooting rate is 95.1%.Described root media is: MS+4.0mg/L IBA+1.0mg/L GA
3+ 0.5mg/L 6-BA+28g/L sucrose+4.0g/L agar, pH is 5.5.
(5) acclimatization and transplants: the blake bottle bottle cap growing to the rooting tube plantlet of 3 ~ 4cm is opened and is placed in natural lighting lower refining seedling after 7 days, test-tube plantlet is taken out from blake bottle, wash root medium off, do not injure root, plant in illumination every day 10 hours in the matrix be mixed into by yellow sand and fiery carbon mud (1:1), intensity of illumination is 4000lx as far as possible, cultivation temperature is 27 DEG C, air humidity is cultivate 7 days in the incubator of 90%, and then transplant planting is in large Tanaka, and transplanting survival rate is 94.2%.
Claims (4)
1. an American red-maple tissue culture and rapid propagation method, is characterized in that comprising the following steps:
(1) explant sterilization: gather the stem section of returning and to rinse after 10 ~ 25min with clear water and brush away impurity above gently with hairbrush; with the 10 ~ 30s that sterilizes in 70% ~ 80% ethanolic solution in superclean bench; afterwards with aseptic washing 3 ~ 5 times; again with 0.1% ~ 0.3% mercuric chloride solution sterilization 10 ~ 25min, with for subsequent use after sucking moisture with aseptic filter paper after aseptic water washing 4 ~ 6 times;
(2) Fiber differentiation: the stem section that step (1) disinfects is cut into band 1 stipes and is about the long stem section of 1.5cm and is inoculated in inducing culture and carry out bud inducement cultivation; first full light culture 5 ~ 10 days under 25 ~ 28 DEG C of conditions after inoculation; then illumination every day is placed in 10 ~ 14 hours; intensity of illumination is 2000 ~ 3000lx, and being placed in cultivation temperature is cultivate until form indefinite bud under the condition of 25 ~ 28 DEG C;
(3) Multiplying culture: step (2) is cultivated the bud seedling obtained; cut bud from base portion and proceed to proliferated culture medium and carry out squamous subculture; first full light culture 2 ~ 3 days under 25 ~ 28 DEG C of conditions after inoculation; then illumination every day is placed in 11 ~ 15 hours; intensity of illumination is 2000 ~ 3000lx; being placed in cultivation temperature is cultivate under the condition of 25 ~ 28 DEG C, and 30 days subcultures once;
(4) culture of rootage: when the bud seedling of step (2) or (3) is grown to 1.5 ~ 2.5cm height; cut into simple bud and be inoculated on root media and carry out culture of rootage; first full light culture 2 ~ 3 days under 25 ~ 28 DEG C of conditions after inoculation; then illumination every day is placed in 14 ~ 16 hours; intensity of illumination is 3000 ~ 5000lx, and cultivation temperature is be cultured under the condition of 25 ~ 28 DEG C to take root;
(5) acclimatization and transplants: the blake bottle bottle cap growing to the rooting tube plantlet of 3 ~ 4cm is opened and is placed in natural lighting lower refining seedling after 5 ~ 7 days, test-tube plantlet is taken out from blake bottle, wash root medium off, do not injure root as far as possible, plant in the matrix be mixed into by yellow sand and fiery carbon mud (1:1) in illumination every day 10 ~ 12 hours, intensity of illumination is 3000 ~ 5000lx, cultivation temperature is 25 ~ 28 DEG C, air humidity is cultivate 5 ~ 7 days in the incubator of 80% ~ 90%, and then transplant planting is in large Tanaka.
2. a kind of American red-maple tissue culture and rapid propagation method according to claim 1, it is characterized in that the inducing culture described in step (2) is: WPM+1.0 ~ 5.0mg/L 6-BA+0.1 ~ 1.0mg/LIBA+15 ~ 30g/L sucrose+3.5 ~ 6.0g/L agar, pH is 5.4 ~ 5.8.
3. a kind of American red-maple tissue culture and rapid propagation method according to claim 1, it is characterized in that the proliferated culture medium described in step (3) is: WPM+0.001 ~ 0.01mg/L TDZ+0.1 ~ 1.0mg/L NAA+15 ~ 30g/L sucrose+3.5 ~ 6.0g/L agar, pH is 5.4 ~ 5.8.
4. a kind of American red-maple tissue culture and rapid propagation method according to claim 1, is characterized in that the root media described in step (4) is: MS+1.0 ~ 5.0mg/L IBA+1.0 ~ 2.0mg/L GA
3+ 0.1 ~ 0.5mg/L 6-BA+15 ~ 30g/L sucrose+3.5 ~ 6.0g/L agar, pH is 5.4 ~ 5.8.
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Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105454048A (en) * | 2015-12-30 | 2016-04-06 | 四川禾木本业农林科技有限公司 | Tissue culture rapid propagation method for acer rubrum |
| CN105918128A (en) * | 2016-05-20 | 2016-09-07 | 芜湖欧标农业发展有限公司 | Method for quickly reproducing acer rubrum |
| CN106069748A (en) * | 2016-06-07 | 2016-11-09 | 江苏东郁园林科技有限公司 | The commercial tissue culture of sprout mating system of American red-maple splendidness in October |
| CN106305427A (en) * | 2016-08-30 | 2017-01-11 | 长江三峡生态园林有限公司 | Tissue culture and rapid propagation method for Acer ginnala |
| CN106472310A (en) * | 2016-10-13 | 2017-03-08 | 李志峰 | A kind of Canada Red maple fast breeding technique and its application |
| CN107278686A (en) * | 2017-06-06 | 2017-10-24 | 山东大丰园农业有限公司 | A kind of American red-maple tissue culture outside sprout-cultivating-bottle radication agent and method for culturing seedlings |
| CN109169282A (en) * | 2018-09-20 | 2019-01-11 | 潍坊职业学院 | A kind of U.S.'s autumn flame Acer palmatum ' Atropurpureum' rapid propagation method |
| CN109247238A (en) * | 2018-11-30 | 2019-01-22 | 四川七彩林业开发有限公司 | A kind of five leaflet maple tissue culture outside sprout-cultivating-bottle radication methods |
| CN109526742A (en) * | 2018-12-25 | 2019-03-29 | 福建农林大学 | A kind of Japanese red maple Green Dragon tissue-cultured seedling subculture multiplication medium formula |
| CN110574688A (en) * | 2019-10-29 | 2019-12-17 | 安徽东方金桥农林科技股份有限公司 | Seedling raising method for North American red maples |
| CN110771506A (en) * | 2019-11-26 | 2020-02-11 | 大连大学 | Method for preparing artificial seeds of red maple |
| CN115885844A (en) * | 2021-08-19 | 2023-04-04 | 大庆石油管理局有限公司 | Autumn flame rapid propagation method after northern domestication |
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Cited By (16)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105454048A (en) * | 2015-12-30 | 2016-04-06 | 四川禾木本业农林科技有限公司 | Tissue culture rapid propagation method for acer rubrum |
| CN105454048B (en) * | 2015-12-30 | 2018-12-21 | 四川七彩林业开发有限公司 | A kind of tissue culture and rapid propagation method of American red-maple |
| CN105918128A (en) * | 2016-05-20 | 2016-09-07 | 芜湖欧标农业发展有限公司 | Method for quickly reproducing acer rubrum |
| CN106069748A (en) * | 2016-06-07 | 2016-11-09 | 江苏东郁园林科技有限公司 | The commercial tissue culture of sprout mating system of American red-maple splendidness in October |
| CN106305427A (en) * | 2016-08-30 | 2017-01-11 | 长江三峡生态园林有限公司 | Tissue culture and rapid propagation method for Acer ginnala |
| CN106305427B (en) * | 2016-08-30 | 2018-04-10 | 长江三峡生态园林有限公司 | A kind of method of Amur maple tissue-culturing quick-propagation |
| CN106472310A (en) * | 2016-10-13 | 2017-03-08 | 李志峰 | A kind of Canada Red maple fast breeding technique and its application |
| CN107278686A (en) * | 2017-06-06 | 2017-10-24 | 山东大丰园农业有限公司 | A kind of American red-maple tissue culture outside sprout-cultivating-bottle radication agent and method for culturing seedlings |
| CN109169282A (en) * | 2018-09-20 | 2019-01-11 | 潍坊职业学院 | A kind of U.S.'s autumn flame Acer palmatum ' Atropurpureum' rapid propagation method |
| CN109247238A (en) * | 2018-11-30 | 2019-01-22 | 四川七彩林业开发有限公司 | A kind of five leaflet maple tissue culture outside sprout-cultivating-bottle radication methods |
| CN109247238B (en) * | 2018-11-30 | 2021-09-24 | 四川七彩林科股份有限公司 | Acer negundo L tissue culture seedling ex-vitro rooting method |
| CN109526742A (en) * | 2018-12-25 | 2019-03-29 | 福建农林大学 | A kind of Japanese red maple Green Dragon tissue-cultured seedling subculture multiplication medium formula |
| CN110574688A (en) * | 2019-10-29 | 2019-12-17 | 安徽东方金桥农林科技股份有限公司 | Seedling raising method for North American red maples |
| CN110771506A (en) * | 2019-11-26 | 2020-02-11 | 大连大学 | Method for preparing artificial seeds of red maple |
| CN110771506B (en) * | 2019-11-26 | 2022-08-05 | 大连大学 | Method for preparing artificial seeds of red maple |
| CN115885844A (en) * | 2021-08-19 | 2023-04-04 | 大庆石油管理局有限公司 | Autumn flame rapid propagation method after northern domestication |
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Application publication date: 20150603 |