CN105010147A - Special culture medium for improving tissue culture propagation speed of haworthia succulent plants and tissue culture method - Google Patents

Special culture medium for improving tissue culture propagation speed of haworthia succulent plants and tissue culture method Download PDF

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CN105010147A
CN105010147A CN201510502501.4A CN201510502501A CN105010147A CN 105010147 A CN105010147 A CN 105010147A CN 201510502501 A CN201510502501 A CN 201510502501A CN 105010147 A CN105010147 A CN 105010147A
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aseptic
tissue culture
culture
medium
propagation speed
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夏时云
韩伟
丁亮
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Hong Ke (tianjin) Agricultural Science And Technology Co Ltd
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Hong Ke (tianjin) Agricultural Science And Technology Co Ltd
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Abstract

The invention provides a special culture medium for improving the tissue culture propagation speed of haworthia succulent plants and a tissue culture method. The special culture medium is characterized in that the ingredients of the special culture medium include a basic culture medium and low-dosage 2,4-D. The tissue culture method comprises the following steps of 1 explant selection and sterilization, 2 inoculation and aseptic strain establishment and 3 transfer and expanding propagation. The problem of low tissue culture propagation speed of the haworthia succulent plants can be solved, the propagation speed of the haworthia plants is remarkably improved, the cost of the whole tissue culture is reduced, and the marketing speed of products is improved.

Description

A kind of special culture media and tissue culture method belonging to succulent tissue culture propagation speed for increasing 12 volumes
Technical field
The present invention relates to the numerous field of biological expansion, mainly relate to a kind of special culture media and the tissue culture method that belong to succulent tissue culture propagation speed for increasing 12 volumes.
Background technology
It is the small-sized succulent of a class perennial herb that 12 volumes belong to (Haworthia Dural), be subordinate to Liliaceae aloe race, in honor of 19 beginning of the century Britain botanist Adrian Hardn Haworth and gaining the name, this genus, by the form of plant and leaf, is divided into sclerophyll class and soft leaf class two class.12 volume platymiscium original producton locations are mostly in South Africa, and Arid Area, China desert also has wild species to distribute, and plant now extensively all over the world.This genus is about divided into 20, various more than 150, horticultural variety and Hybrid various, price is not from a few unit to unit up to ten thousand etc.12 volume platymisciums are leaf to be full of variety, and beautiful appearance, has very high ornamental value, and many types habit is strong, cultivates less demanding, is one of most popular at present decorative indoor plant of China.
12 volume platymiscium Traditional breeding processes mainly contain cuttage, plant division and sowing etc.Cuttage survival rate is extremely low, is applicable to plant fan and carries out small-scale breeding to plant, be not suitable for commercial large-scale breeding.Plant division is that 12 volumes comparatively commonly used belong to Traditional breeding processes, but annual breeding amount only 3-5 strain, famous and precious kind is less, and reproduction rate is extremely low.Seeding method is subject to 12 volumes and belongs to the few restriction of seed amount, and meanwhile, percentage of seedgermination is extremely low again, and even the longer time can grow up to need 2 years, and owing to being sexual propagation, Character instability, does not meet the requirement of commodity production.Above-mentioned factor makes 12 volume platymisciums hold at high price, and ordinary people can not consumed.
Tissue culture propagation method is Recent study and 12 conventional volume platymiscium propagation methods, can acquired character stablize, aseptic seedling, comparatively Traditional breeding processes, reproduction rate increases, but still it is slow to there is tissue culture propagation speed, survival rate is not high, the problem that value added on products is few.In order to solve the low problem of above-mentioned 12 volume platymiscium tissue culture propagation rates, the invention provides medium and tissue culture method that a kind of increase by 12 volumes belong to succulent tissue culture propagation speed, the reproductive speed of 12 volume platymisciums can be significantly improved, obtain a large amount of 12 volumes in a short time and belong to succulent plant shoots.
Summary of the invention
For solving the low problem of the tissue culture propagation speed that exists in above-mentioned background technology, the invention provides a kind of special culture media belonging to succulent tissue culture propagation speed for increasing 12 volumes, medium component is: 2,4-D of basal medium and low dosage; Described basic media components is: MS20g, carragheen 1g, 6-BA (6-benzyl aminoadenine) 0.1mg, KT (CPPU) 0.5mg., described 2,4-D is 2,4-dichlorphenoxyacetic acid, and content range is 0.1-0.25mg/L.
A kind of special culture media tissue culture method belonging to succulent tissue culture propagation speed for increasing 12 volumes realizes as follows:
(1) selection of explant and sterilization
Getting grows well waits the blade (or lateral bud) organizing the plant trained, and carries out disinfection, is cut into 1 × 1cm 2size, the blade that the back side scratches, to be seeded.
(2) foundation of inoculation and aseptic strain
Take the photograph son by the blade inoculation cut to (the direct oblique cutting of lateral bud enters, and top upwards) on aseptic culture medium with aseptic, blade lies in be cultivated on basal plane, medium is contacted under streaking that side direction of wound, after 1-2 week, external body expands rapidly, and part edge differentiates seedling, obtains aseptic strain.
(3) switching is numerous with expansion
By the aseptic strain obtained, be cut into segment, be seeded on proliferated culture medium of the present invention, carry out Multiplying culture, after 2-3 week, cut single for large plant, put on strong seedling culture base or root media, continue to cultivate 2-3 week, plant can be taken out, wash out medium, put into shady and cool place and dry 1-2 days, be transplanted in compost, 12 volume class plant shoots can be obtained.
Sterilization method in described step (1) is: rinse in the blade chosen or lateral bud clear water, then puts into the carbendazim solution that mass content is about 5% and soak 10-15 minute, then rinses well with clear water; Superclean bench is put into aseptic bottle 1% raw mercury process 3-5 minute (determining soak time length according to the tender degree of children and pollution level), pours out raw mercury, with aseptic water washing 3-6 time of cooling.Blade (or lateral bud) cutting process need be put in sterile petri dish, and adopts sterile razor blade.
In described step (2), aseptic culture medium is placed in aseptic culture medium bottle, its composition is: the condition of culture that in MS20g, carragheen 1g, 6-BA0.1mg, KT0.5mg, aseptic strain is set up is: tightened by the aseptic culture medium bottle inoculated, be put in illumination box and cultivate, the temperature controlling incubator is 25 degrees Celsius, and light application time is 16 hours/day.
Multiplying culture process in described step (3) can repeat, and can will breed the tissue that or plant is inoculated on proliferated culture medium of the present invention as aseptic strain, can obtain 12 a large amount of volume class seedling in a short time.
Compared with prior art, the invention has the advantages that:
1, in succulent tissue culture propagation process, with the addition of 2 of low dosage in the medium first, 4-D, in the Multiplying culture stage, a large amount of buds and callus can be produced simultaneously, doubly, the numerous and differentiation that is bud of the expansion for later callus provides a large amount of basic materials to 1.5-2.5 when to increase numerous amount be the medium not adding 2,4-D;
2, the present invention significantly can increase the reproduction speed that 12 volumes belong to succulent, for reducing whole group of training cost, improving launch speed, having established solid foundation.
Embodiment
Embodiment 1:
1. the selection of explant and sterilization:
Get good treating of growing to rinse in the blade clear water of tissue culture plant, then put into the carbendazim solution that mass content is about 5% and soak 10-15 minute, then rinse well with clear water.
Superclean bench is put into aseptic bottle 1% mercuric chloride solution process 3-5 minute (determining soak time length according to the tender degree of children and pollution level), pour out mercuric chloride solution, with aseptic water washing 3-6 time of cooling, be put into sterile petri dish, be cut into 1 × 1cm by sterile razor blade 2the blade of size, and vacuum side of blade is scratched, to be seeded.
2. the foundation of inoculation and aseptic strain:
With the aseptic blade taken the photograph son and will cut, lie on aseptic culture basal plane, contact medium under streaking that side direction of wound, tighten aseptic culture medium bottleneck, carry out mark, put into illumination box to cultivate, regulate temperature to be 25 degrees Celsius, light application time is 16 hours/day, cultivate 1-2 week, explant expands rapidly, produces callus, and differentiates seedling at part edge.
3. switching is numerous with expansion:
The explant of differentiation is taken out, is cut into segment, is seeded on special culture media of the present invention, wherein 2,4-D content are 0.1mg/L, after continuing to cultivate 2-3 week, can obtain a large amount of callus and aseptic Multiple Buds, proliferative amount is 1.7 times of the medium not adding 2,4-D.Repeat above process, callus and Multiple Buds quantity increase sharply.Now choose that large sprout is single cuts, put on strong seedling culture base or root media, after cultivating 2-3 week, plant is taken out, washes away medium, put into shady and cool place and dry 1-2 days, can be transplanted in compost, obtain 12 volumes and belong to succulent seedling.
Embodiment 2
1. the selection of explant and sterilization:
Get good treating of growing to rinse in the blade clear water of tissue culture plant, then put into the carbendazim solution that mass content is about 5% and soak 10-15 minute, then rinse well with clear water.
Superclean bench is put into aseptic bottle 1% mercuric chloride solution process 3-5 minute (determining soak time length according to the tender degree of children and pollution level), pour out mercuric chloride solution, with aseptic water washing 3-6 time of cooling, be put into sterile petri dish, be cut into 1 × 1cm by sterile razor blade 2the blade of size, and vacuum side of blade is scratched, to be seeded.
2. the foundation of inoculation and aseptic strain:
With the aseptic blade taken the photograph son and will cut, lie on aseptic culture basal plane, contact medium under streaking that side direction of wound, tighten aseptic culture medium bottleneck, carry out mark, put into illumination box to cultivate, regulate temperature to be 25 degrees Celsius, light application time is 16 hours/day, cultivate 1-2 week, explant expands rapidly, produces callus, and differentiates seedling at part edge.
3. switching is numerous with expansion:
Taken out by the explant of differentiation, be cut into segment, be seeded on special culture media of the present invention, wherein 2,4-D content are 0.15mg/L, after continuing to cultivate 2-3 week, can obtain a large amount of callus and aseptic Multiple Buds, and proliferative amount is 1.9 times not containing 2,4-D.Repeat above process, Multiple Buds quantity increases sharply.Now choose that large sprout is single cuts, put on strong seedling culture base or root media, after cultivating 2-3 week, plant is taken out, washes away medium, put into shady and cool place and dry 1-2 days, can be transplanted in compost, obtain 12 volumes and belong to succulent seedling.
Embodiment 3
1. the selection of explant and sterilization:
Get good treating of growing to rinse in the blade clear water of tissue culture plant, then put into the carbendazim solution that mass content is about 5% and soak 10-15 minute, then rinse well with clear water.
Superclean bench is put into aseptic bottle 1% mercuric chloride solution process 3-5 minute (determining soak time length according to the tender degree of children and pollution level), pour out mercuric chloride solution, with aseptic water washing 3-6 time of cooling, be put into sterile petri dish, be cut into 1 × 1cm by sterile razor blade 2the blade of size, and vacuum side of blade is scratched, to be seeded.
2. the foundation of inoculation and aseptic strain:
With the aseptic blade taken the photograph son and will cut, lie on aseptic culture basal plane, contact medium under streaking that side direction of wound, tighten aseptic culture medium bottleneck, carry out mark, put into illumination box to cultivate, regulate temperature to be 25 degrees Celsius, light application time is 16 hours/day, cultivate 1-2 week, explant expands rapidly, produces callus, and differentiates seedling at part edge.
3. switching is numerous with expansion:
Taken out by the explant of differentiation, be cut into segment, be seeded on special culture media of the present invention, wherein 2,4-D content are 0.25mg/L, after continuing to cultivate 2-3 week, can obtain a large amount of callus and aseptic Multiple Buds, and proliferative amount is not containing 2.3 times of 2,4-D.Repeat above process, Multiple Buds quantity increases sharply.Now choose that large sprout is single cuts, put on strong seedling culture base or root media, after cultivating 2-3 week, plant is taken out, washes away medium, put into shady and cool place and dry 1-2 days, can be transplanted in compost, obtain 12 volumes and belong to succulent seedling.
Embodiment 4:
1. the selection of explant and sterilization:
Getting grows good treat that tissue culture plant lateral bud is put in clear water and rinses, then puts into the carbendazim solution that mass content is about 5% and soak 10-15 minute, then rinses well with clear water.
Superclean bench is put into aseptic bottle and falls 1% mercuric chloride solution process 3-5 minute (determining soak time length according to the tender degree of children and pollution level), pour out mercuric chloride solution, with aseptic water washing 3-6 time of cooling, to be seeded.
2. the foundation of inoculation and aseptic strain:
Enter on aseptic culture medium with aseptic son of taking the photograph by the direct oblique cutting of lateral bud after sterilization, top upwards, is tightened aseptic culture medium bottleneck, is carried out mark, and put into illumination box and cultivate, control temperature is 25 degrees Celsius, and light application time is 16 hours/day.Cultivate 1-2 week, explant expands rapidly, obtains callus and Multiple Buds.
3. switching is numerous with expansion:
The explant of differentiation is taken out, is cut into segment, is seeded on special culture media of the present invention, wherein 2,4-D content are 0.1mg/L, after continuing to cultivate 2-3 week, can obtain a large amount of callus and aseptic Multiple Buds, value added on products is 2.2 times not containing 2,4-D medium proliferative amount.Repeat above process, Multiple Buds quantity increases sharply.Now choose that large sprout is single cuts, put on strong seedling culture base or root media, after cultivating 2-3 week, plant can be taken out, wash out medium, put into shady and cool place and dry 1-2 days, can be transplanted in compost, obtain 12 volumes and belong to succulent seedling.
Embodiment 5:
1. the selection of explant and sterilization:
Getting grows good treat that tissue culture plant lateral bud is put in clear water and rinses, then puts into the carbendazim solution that mass content is about 5% and soak 10-15 minute, then rinses well with clear water.
Superclean bench is put into aseptic bottle and falls 1% mercuric chloride solution process 3-5 minute (determining soak time length according to the tender degree of children and pollution level), pour out mercuric chloride solution, with aseptic water washing 3-6 time of cooling, to be seeded.
2. the foundation of inoculation and aseptic strain:
Enter on aseptic culture medium with aseptic son of taking the photograph by the direct oblique cutting of lateral bud after sterilization, top upwards, is tightened aseptic culture medium bottleneck, is carried out mark, and put into illumination box and cultivate, control temperature is 25 degrees Celsius, and light application time is 16 hours/day.Cultivate 1-2 week, explant expands rapidly, obtains callus and Multiple Buds.
3. switching is numerous with expansion:
The explant of differentiation is taken out, be cut into segment, be seeded on special culture media of the present invention, wherein 2,4-D content are 0.15mg/L, after continuing to cultivate 2-3 week, can obtain aseptic Multiple Buds, repeat above process, callus and Multiple Buds quantity increase sharply, proliferative amount is 2.5 times that do not add 2,4-D medium proliferative amounts.Now choose that large sprout is single cuts, put on strong seedling culture base or root media, after cultivating 2-3 week, plant can be taken out, wash out medium, put into shady and cool place and dry 1-2 days, can be transplanted in compost, obtain 12 volumes and belong to succulent seedling.
Embodiment 6:
1. the selection of explant and sterilization:
Getting grows good treat that tissue culture plant lateral bud is put in clear water and rinses, then puts into the carbendazim solution that mass content is about 5% and soak 10-15 minute, then rinses well with clear water.
Superclean bench is put into aseptic bottle and falls 1% mercuric chloride solution process 3-5 minute (determining soak time length according to the tender degree of children and pollution level), pour out mercuric chloride solution, with aseptic water washing 3-6 time of cooling, to be seeded.
2. the foundation of inoculation and aseptic strain:
Enter on aseptic culture medium with aseptic son of taking the photograph by the direct oblique cutting of lateral bud after sterilization, top upwards, is tightened aseptic culture medium bottleneck, is carried out mark, and put into illumination box and cultivate, control temperature is 25 degrees Celsius, and light application time is 16 hours/day.Cultivate 1-2 week, explant expands rapidly, obtains callus and Multiple Buds.
3. switching is numerous with expansion:
The explant of differentiation is taken out, be cut into segment, be seeded on special culture media of the present invention, wherein 2,4-D content are 0.25mg/L, after continuing to cultivate 2-3 week, can obtain aseptic Multiple Buds, repeat above process, callus and Multiple Buds quantity increase sharply, proliferative amount is 2.1 times that do not add 2,4-D medium.Now choose that large sprout is single cuts, put on strong seedling culture base or root media, after cultivating 2-3 week, plant can be taken out, wash out medium, put into shady and cool place and dry 1-2 days, can be transplanted in compost, obtain 12 volumes and belong to succulent seedling.

Claims (6)

1. belong to special culture media and the tissue culture method of succulent tissue culture propagation speed for increasing 12 volumes, it is characterized in that the composition of this special culture media is: 2,4-D of basal medium and low dosage;
Described basic media components is: MS20g, carragheen 1g, 6-BA (6-benzyl aminoadenine) 0.1mg, KT (CPPU) 0.5mg;
Described 2,4-D is 2,4-dichlorphenoxyacetic acid, and content range is 0.1-0.25mg/L.
2. a kind of special culture media and tissue culture method belonging to succulent tissue culture propagation speed for increasing 12 volumes according to claim 1, is characterized in that, be through following steps:
(1) selection of explant and sterilization
Getting grows well waits the blade (or lateral bud) organizing the plant trained, and carries out disinfection, is cut into 1 × 1cm 2size, the blade that the back side scratches, to be seeded;
(2) foundation of inoculation and aseptic strain
With aseptic take the photograph son by the blade inoculation cut, on aseptic culture medium, (the direct oblique cutting of lateral bud enters, top upwards), blade lies on aseptic culture basal plane, medium is contacted under streaking that side direction of wound, after 1-2 week, external body expands rapidly, part edge differentiates seedling, obtains aseptic strain;
(3) switching is numerous with expansion
By the aseptic strain obtained, be cut into segment, be seeded on proliferated culture medium of the present invention, containing 2,4-D of 0.1-0.25mg/L, after carrying out Multiplying culture 2-3 week, cut single for large plant, put on strong seedling culture base or root media, continue to cultivate 2-3 week, plant can be taken out, wash out medium, put into shady and cool place and dry 1-2 days, be transplanted in compost, 12 volumes can be obtained and belong to plant shoots.
3. a kind of special culture media and tissue culture method belonging to succulent tissue culture propagation speed for increasing 12 volumes according to claim 2, it is characterized in that, sterilization method in described step (1) is: rinse in the blade chosen or lateral bud clear water, put into the carbendazim solution that content is about 5% and soak 10-15 minute, then rinse well with clear water; Superclean bench being put into aseptic bottle 1% mercuric chloride solution process 3-5 minute (determining soak time length according to the tender degree of children and pollution level), pouring out mercuric chloride solution, with cooling aseptic water washing 3-6 time.Blade (or lateral bud) cutting process need be put in sterile petri dish, and adopts sterile razor blade.
4. a kind of special culture media and tissue culture method belonging to succulent tissue culture propagation speed for increasing 12 volumes according to claim 2, it is characterized in that, the aseptic culture medium in described step (2) is the basal medium after sterilizing.
5. a kind of special culture media and tissue culture method belonging to succulent tissue culture propagation speed for increasing 12 volumes according to claim 2, it is characterized in that, the condition of culture that aseptic strain in described step (2) is set up is: tightened by the aseptic culture medium bottle inoculated, be put in illumination box and cultivate, the temperature controlling incubator is 25 degrees Celsius, and light application time is 16 hours/day.
6. a kind of special culture media and tissue culture method belonging to succulent tissue culture propagation speed for increasing 12 volumes according to claim 2, it is characterized in that, Multiplying culture process in described step (3) can repeat, can will breed the tissue that or plant is inoculated on proliferated culture medium of the present invention as aseptic strain, carry out squamous subculture.
CN201510502501.4A 2015-08-14 2015-08-14 Special culture medium for improving tissue culture propagation speed of haworthia succulent plants and tissue culture method Pending CN105010147A (en)

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CN105766635A (en) * 2016-03-14 2016-07-20 龙岩市禾康生物科技有限公司 Method for tissue culture and rapid propagation of succulent plants
CN106305417A (en) * 2016-08-19 2017-01-11 天津市园林花卉示范中心 Haworthia truncata ex-vitro rooting tissue culture seedling growing method
CN107027632A (en) * 2017-06-08 2017-08-11 北京农学院 Everything rapid propagation in vitro method by explant of blade
CN107912158A (en) * 2017-11-15 2018-04-17 广西壮族自治区农业科学院花卉研究所 A kind of blade section cottage method of Bryophyllum succulent
CN108719065A (en) * 2018-05-21 2018-11-02 句容市茂润苗木有限公司 A kind of succulent rapid propagation method
CN113826553A (en) * 2021-10-28 2021-12-24 江苏农林职业技术学院 Method for culturing and rapidly propagating succulent plant Tianzhuang pancakea tissues

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105638463A (en) * 2015-12-30 2016-06-08 四川禾木本业农林科技有限公司 Tissue-culture rapid propagation method for succulents
CN105766635A (en) * 2016-03-14 2016-07-20 龙岩市禾康生物科技有限公司 Method for tissue culture and rapid propagation of succulent plants
CN105766635B (en) * 2016-03-14 2017-12-22 龙岩市禾康生物科技有限公司 A kind of method of succulent tissue-culturing rapid propagation
CN106305417A (en) * 2016-08-19 2017-01-11 天津市园林花卉示范中心 Haworthia truncata ex-vitro rooting tissue culture seedling growing method
CN107027632A (en) * 2017-06-08 2017-08-11 北京农学院 Everything rapid propagation in vitro method by explant of blade
CN107912158A (en) * 2017-11-15 2018-04-17 广西壮族自治区农业科学院花卉研究所 A kind of blade section cottage method of Bryophyllum succulent
CN108719065A (en) * 2018-05-21 2018-11-02 句容市茂润苗木有限公司 A kind of succulent rapid propagation method
CN113826553A (en) * 2021-10-28 2021-12-24 江苏农林职业技术学院 Method for culturing and rapidly propagating succulent plant Tianzhuang pancakea tissues

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Application publication date: 20151104