CN103444540A - Method for quickly breeding plumeria rubra by tissue culture - Google Patents
Method for quickly breeding plumeria rubra by tissue culture Download PDFInfo
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- CN103444540A CN103444540A CN2013104117753A CN201310411775A CN103444540A CN 103444540 A CN103444540 A CN 103444540A CN 2013104117753 A CN2013104117753 A CN 2013104117753A CN 201310411775 A CN201310411775 A CN 201310411775A CN 103444540 A CN103444540 A CN 103444540A
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Abstract
The invention provides a method for quickly breeding plumeria rubra by tissue culture. The method comprises the following steps of carrying out tissue culture and rapid propagation with MS (Murashige and Skoog) as a minimal medium and plumeria rubra leaves as explants under different hormone components and concentration levels; carrying out sterilization treatment and inoculation induction to form a callus tissue, and then carrying out callus tissue differentiation, bud multiplication and rooting culture to form a complete plant seedling; and finally carrying out acclimatization and transplantation, wherein the minimal medium is based on the MS culture medium, and supplemented with components including 6-benzyl aminoadenine, 2,4-dichlorphenoxyacetic acid, naphthylacetic acid, sucrose, active carbon and the like. According to the invention, tissue culture is applied to overcome the problem that the plumeria rubra is too long in seedling-raising period and low in propagation coefficient, so that factory-like quick seedling raising can be carried out for meeting the demands of the market on the plumeria rubra.
Description
Technical field
The present invention relates to a kind of tissue culture rapid speed and breed the method for safflower frangipanis.
Background technology
Frangipanis, the defoliation small arbor of Apocynaceae, have another name called remote cape jasmine, originates in the american torrid zone areas such as Mexico, imports China into from Burma etc. at first.Its petal is pure white, fickle in love yellowish, and the utmost point yolk like protein encapsulation, hence obtains one's name.Between every year 4, May, dignified graceful frangipanis just bursts forth successively, gives off a strong fragrance, and moves one deeply, and 5 film clips lobes are folded and given birth to, and the spy resembles the pinwheel of children's record book in accordion form.In fact, frangipanis, except white, has red, yellow two kinds in addition, all can extract essence for manufacture superior cosmetics, perfumed soap and food additives, and price is quite high, has the business development potentiality; Also fresh flower can be dried afterwards for what make tea, be commonly called as egg jasmine tea, the effect of controlling hot diarrhea, moistening lung removing toxic substances is arranged.Frangipanis is tree-like attractive in appearance, and the stem multi-branched is extraordinary-looking, in different poses and with different expressions; Leaf, like loquat, after falling winter, just stays semicircular leaf scar on end of the branch, quite resemble the deer horn that is embroidered with beautiful spot, is that torrid areas afforestation, garden are arranged, the potted plant first-selected dungarunga good merchantable brand of viewing and admiring.Bark is thin and be celadon, is rich in poisonous white juice, can be used to external application, cures the diseases such as scabies, redness.Timber white, light weight and soft, can musical instrument processed, tableware or furniture.Frangipanis is loved by the people in a lot of areas and country.In Laos, frangipanis is decided to be national flower and enjoys worships; In Hawaii, tropical tourist attraction, people like conspiring to create garland as the ornament of wearing using adopting the frangipanis got off, so frangipanis is again to signify Hawaiian red-letter day; Frangipanis is decided to be one of " five tree six flowers " and extensively plants by Buddhist temple, therefore have another name called " mausoleum tree " or " tower tree ".In China, frangipanis is not only the city flower of Zhaoqing City of Guangdong Province, and Xishuangbanna people from Dai Nationality of enthusiasm receives the best speciality of guests especially.
Simultaneously, the kind of frangipanis has three kinds: chrysanthemum frangipanis, pink blossom frangipanis and safflower frangipanis.Chrysanthemum frangipanis and pink blossom frangipanis are more common at home, and the safflower frangipanis is quite rare, also very famous and precious! Breeding of rose Plumeria rubra be take cuttage as main now, easily survives and transplants; Also can breed with planting seed; Carry out grafting with domestic common chrysanthemum frangipanis as large tree stock, can accelerate the Landscape Application in the safflower frangipanis area such as in south China.But such reproduction speed is but quite slow, the cycle is also long, can not meet the demand of domestic market to safflower frangipanis seedling far away.Therefore, the method that adopts tissue to cultivate is to accelerate the effective way of its breeding.
Summary of the invention
The objective of the invention is to cultivate the method for carrying out safflower frangipanis Fast-propagation seedling by tissue.
Technical scheme of the present invention is:
A kind of tissue culture rapid speed is bred the method for safflower frangipanis, it is characterized in that: take MS as minimal medium, safflower frangipanis blade is that explant carries out tissue-culturing rapid propagation, after sterile-processed, inoculation is induced and is formed callus, again through Calli Differentiation, bud propagation and culture of rootage, form the whole plant seedling, finally carry out hardening and transplanting.
Described tissue culture rapid speed is bred the method for safflower frangipanis, comprises the following steps:
The selection of A, explant and disinfecting: after choosing the blade on the safflower frangipanis maternal plant of well-grown stalwartness, be put in temporarily the place two days in indoor cool place, after the juice discharged in blade is dry, after cleaning 10 minutes with running water, on aseptic super-clean bench, dry, then, blade is cut into to each fritter of 2 centimetres of length and width, fritter is put into to 75% alcohol rinsing once (in 10 seconds), proceed in 0.1% mercuric chloride solution (being added with five or six of Tween-20s) and sterilize 10 minutes, with aseptic water washing 3 ~ 4 times, cut limb standby;
B, explant induction: will be seeded on callus inducing medium through fritter standby in steps A, incubation time is 40 ~ 50 days, in the incision of blade, expands and forms jade-green callus.
C, indefinite bud induce differentiation: the callus obtained in step B is inoculated on the medium of adventitious bud inducing, cultivates 32 ~ 35 days, crowd shoots occurs;
D, culture of rootage: the crowd shoots plant division by obtaining in step C, proceed in root media, to cultivate 30 ~ 33 days, the seedling base portion short of white occurs and grows complete root system;
E, hardening and transplanting: will obtain the bottle seedling of complete plantlet in step D, be put in the inside, greenhouse hardening 5 days, the bottle seedling is opened to hardening 2 days, then from bottle, remove plant, carefully wash away the agar medium adhered to, carry out at ambient temperature the Nutrition Soil cultivation.
Described explant is disinfected to adopt in 75% alcohol-pickled sterilizing 5 ~ 10 seconds and 0.1 ~ 0.2% mercuric chloride (being added with several of Tween-80s) and is soaked sterilizing dual twice sterilization method of 5 ~ 10 minutes.
Described medium of inducing with Callus of Leaf is MS, 6-BA 0 ~ 3 mg/L, 2.4-D 0 ~ 0.5 mg/L, NAA 0 ~ 1mg/L, wherein: MS is conventional minimal medium Murashige&Skoog 1962,6-BA is 6-benzyl aminoadenine, 2,4-D is 2, the 4-dichlorphenoxyacetic acid, NAA is methyl α-naphthyl acetate.
Described adventitious bud inducing differential medium is: MS, 6-BA 1 ~ 3 mg/L, NAA 0.1 ~ 0.5 mg/L, and wherein: MS is conventional minimal medium Murashige&Skoog 1962, and 6-BA is 6-benzyl aminoadenine, and NAA is methyl α-naphthyl acetate.
Described root media is: 1/2MS, NAA 0.01 ~ 0.1mg/L, 2 ~ 3% sucrose, 0.8 ~ 1.0% agar, wherein: MS is conventional minimal medium Murashige&Skoog 1962, macroelement in this routine minimal medium is reduced by half and obtains 1/2MS, and NAA is methyl α-naphthyl acetate.
Described explant induction medium and adventitious bud inducing differential medium all are incorporated as 1.0% agar, 2 ~ 3% sucrose, 0.5% active carbon and natural organic additive.
Described natural organic additive is 0.2% coconut milk, 0.1% apple juice, 0.1% banana puree mixture
The invention has the advantages that: the present invention utilizes tissue to cultivate to carry out Vitro Quick Reproduction significant at following three aspects:: (1) reproduction speed is fast, and climate factor impact, all can breed indoor throughout the year, starts fast; (2) be convenient to keep good strains of seeds, synchronism is good; (3) very important meaning is arranged preserving and breed aspect good mutant or the good first generation of hybrid; (4) demand of satisfying the market to safflower frangipanis seedling rapidly, but make safflower frangipanis spread after tissue is cultivated.
The present invention's medium Induction time used is short, it is fast to emerge, and the propagation frequency is high, especially emerges neat, and easy operating, carry out large-scale production.
The present invention is conceived to organize the practical production of training company carry out, what aspect the drawing materials of explant, use is blade, the petiole of wide material sources, the medium used is all minimal medium MS, the hormone of use be all on market, easily buy obtain, cheap (as 6-BA, NAA, 2.4-D etc.).
The present invention adopts method for tissue culture first, carry out the work of safflower frangipanis seedling Fast-propagation, this provides a new way for safflower frangipanis factorial seedling growth, and the artificial propagation that can carry out the safflower frangipanis, the method for tissue culture that germ plasm resource is preserved in vitro and utilized are provided.
Specific implementation method
Embodiment 1
A kind of tissue culture rapid speed is bred the method for safflower frangipanis, comprises the following steps:
The selection of A, explant and disinfecting: after choosing the blade on the safflower frangipanis maternal plant of well-grown stalwartness, be put in temporarily the place two days in indoor cool place, after the juice discharged in blade is dry, after cleaning 10 minutes with running water, on aseptic super-clean bench, dry, then, blade is cut into to each fritter of 2 centimetres of length and width, fritter is put into to 75% alcohol rinsing once (5 second), proceed in 0.1% mercuric chloride solution (being added with five of Tween-20s) and sterilize 10 minutes, with aseptic water washing 3 times, cut limb standby;
B, explant induction: will be through fritter standby in steps A, be seeded on callus inducing medium, this medium is: MS+6-BA 3.0 mg/L+2,4-D 0.1 mg/L+NAA 1 mg/L, add 1.0% agar, 3% sucrose, 0.5% active carbon and natural organic additive in medium.Incubation time is 44 days, in the incision of blade, expands and forms jade-green callus.
Described superclean bench, model is HS-1300, by SuZhou Antai Air Tech Co., Ltd. of Su Jing group, is produced;
C, indefinite bud induce differentiation: the callus obtained in step B is inoculated on the medium of adventitious bud inducing, this medium is: MS+6-BA 2.0 mg/L+NAA 0.5mg/L adds 1.0% agar, 3% sucrose, 0.5% active carbon and natural organic additive in medium.Cultivate 32 days, crowd shoots occurs;
D, culture of rootage: by the crowd shoots plant division obtained in step C, proceed in root media, this medium is: MS+NAA 0.02 mg/L adds 1.0% agar, 2% sucrose and 0.5% active carbon in medium, cultivate 30 days, the seedling base portion short of white occurs and grows complete root system;
E, hardening and transplanting: will obtain the bottle seedling of complete plantlet in step D, be put in the inside, greenhouse hardening 5 days, the bottle seedling is opened to hardening 2 days, then from bottle, remove plant, carefully wash away the agar medium adhered to, carry out at ambient temperature the Nutrition Soil cultivation.
Embodiment 2
A kind of tissue culture rapid speed is bred the method for safflower frangipanis, comprises the following steps:
The selection of A, explant and disinfecting: after choosing the blade on the safflower frangipanis maternal plant of well-grown stalwartness, be put in temporarily the place two days in indoor cool place, after the juice discharged in blade is dry, after cleaning 10 minutes with running water, on aseptic super-clean bench, dry, then, blade is cut into to each fritter of 3 centimetres of length and width, fritter is put into to 75% alcohol rinsing once (10 second), proceed in 0.2% mercuric chloride solution (being added with six of Tween-20s) and sterilize 5 minutes, with aseptic water washing 4 times, cut limb standby;
B, explant induction: will be through fritter standby in steps A, be seeded on callus inducing medium, this medium is: MS+6-BA 2.0 mg/L+2,4-D 0.5 mg/L+NAA 0.5 mg/L, add 1.0% agar, 3% sucrose, 0.5% active carbon and natural organic additive in medium.Incubation time is 40 days, in the incision of blade, expands and forms jade-green callus.
Described superclean bench, model is HS-1300, by SuZhou Antai Air Tech Co., Ltd. of Su Jing group, is produced;
C, indefinite bud induce differentiation: the callus obtained in step B is inoculated on the medium of adventitious bud inducing, this medium is: MS+6-BA 2.0 mg/L+NAA 0.2mg/L adds 1.0% agar, 3% sucrose, 0.5% active carbon and natural organic additive in medium.Cultivate 30 days, crowd shoots occurs;
D, culture of rootage: by the crowd shoots plant division obtained in step C, proceed in root media, this medium is: MS+NAA 0.01 mg/L, the agar, 3% sucrose and 0.5% the active carbon that add 0.8 % in medium, cultivate 34 days, the seedling base portion short of white occurs and grows complete root system;
E, hardening and transplanting: will obtain the bottle seedling of complete plantlet in step D, be put in the inside, greenhouse hardening 5 days, the bottle seedling is opened to hardening 2 days, then from bottle, remove plant, carefully wash away the agar medium adhered to, carry out at ambient temperature the Nutrition Soil cultivation.
Embodiment 3
A kind of tissue culture rapid speed is bred the method for safflower frangipanis, comprises the following steps:
The selection of A, explant and disinfecting: after choosing the blade on the safflower frangipanis maternal plant of well-grown stalwartness, be put in temporarily the place two days in indoor cool place, after the juice discharged in blade is dry, after cleaning 10 minutes with running water, on aseptic super-clean bench, dry, then, blade is cut into to each fritter of 3 centimetres of length and width, fritter is put into to 75% alcohol rinsing once (8 second), proceed in 0.2% mercuric chloride solution (being added with five of Tween-20s) and sterilize 8 minutes, with aseptic water washing 4 times, cut limb standby;
B, explant induction: will be through fritter standby in steps A, be seeded on callus inducing medium, this medium is: MS+6-BA 2 mg/L+2,4-D 0.1 mg/L+NAA 0.5 mg/L, add 1.0% agar, 3% sucrose, 0.5% active carbon and natural organic additive in medium.Incubation time is 48 days, in the incision of blade, expands and forms jade-green callus.
Described superclean bench, model is HS-1300, by SuZhou Antai Air Tech Co., Ltd. of Su Jing group, is produced;
C, indefinite bud induce differentiation: the callus obtained in step B is inoculated on the medium of adventitious bud inducing, this medium is: MS+6-BA 1.0 mg/L+NAA 0.2mg/L adds 1.0% agar, 3% sucrose, 0.5% active carbon and natural organic additive in medium.Cultivate 35 days, crowd shoots occurs;
D, culture of rootage: by the crowd shoots plant division obtained in step C, proceed in root media, this medium is: MS, do not add any hormone, add 0.8% agar, 2% sucrose and 0.5% active carbon in medium, cultivate 33 days, the seedling base portion short of white occurs and grows complete root system;
E, hardening and transplanting: will obtain the bottle seedling of complete plantlet in step D, be put in the inside, greenhouse hardening 5 days, the bottle seedling is opened to hardening 2 days, then from bottle, remove plant, carefully wash away the agar medium adhered to, carry out at ambient temperature the Nutrition Soil cultivation.
Embodiment 4
A kind of tissue culture rapid speed is bred the method for safflower frangipanis, comprises the following steps:
The selection of A, explant and disinfecting: after choosing the blade on the safflower frangipanis maternal plant of well-grown stalwartness, be put in temporarily the place two days in indoor cool place, after the juice discharged in blade is dry, after cleaning 10 minutes with running water, on aseptic super-clean bench, dry, then, blade is cut into to each fritter of 2 centimetres of length and width, fritter is put into to 75% alcohol rinsing once (6 second), proceed in 0.1% mercuric chloride solution (being added with five of Tween-20s) and sterilize 8 minutes, with aseptic water washing 3 times, cut limb standby;
B, explant induction: will be through fritter standby in steps A, be seeded on callus inducing medium, this medium is: MS+6-BA 3.0 mg/L+2,4-D 0.5 mg/L+NAA 0.5 mg/L, add 1.0% agar, 3% sucrose, 0.5% active carbon and natural organic additive in medium.Incubation time is 42 days, in the incision of blade, expands and forms jade-green callus.
Described superclean bench, model is HS-1300, by SuZhou Antai Air Tech Co., Ltd. of Su Jing group, is produced;
C, indefinite bud induce differentiation: the callus obtained in step B is inoculated on the medium of adventitious bud inducing, this medium is: MS+6-BA 2.0 mg/L+NAA 0.4 mg/L adds 1.0% agar, 3% sucrose, 0.5% active carbon and natural organic additive in medium.Cultivate 34 days, crowd shoots occurs;
D, culture of rootage: by the crowd shoots plant division obtained in step C, proceed in root media, this medium is: MS+NAA 0.1 mg/L adds 1.0% agar, 3% sucrose and 0.5% active carbon in medium, cultivate 30 days, the seedling base portion short of white occurs and grows complete root system;
E, hardening and transplanting: will obtain the bottle seedling of complete plantlet in step D, be put in the inside, greenhouse hardening 5 days, the bottle seedling is opened to hardening 2 days, then from bottle, remove plant, carefully wash away the agar medium adhered to, carry out at ambient temperature the Nutrition Soil cultivation.
Embodiment 5
A kind of tissue culture rapid speed is bred the method for safflower frangipanis, comprises the following steps:
The selection of A, explant and disinfecting: after choosing the blade on the safflower frangipanis maternal plant of well-grown stalwartness, be put in temporarily the place two days in indoor cool place, after the juice discharged in blade is dry, after cleaning 10 minutes with running water, on aseptic super-clean bench, dry, then, blade is cut into to each fritter of 3 centimetres of length and width, fritter is put into to 75% alcohol rinsing once (10 second), proceed in 0.2% mercuric chloride solution (being added with five of Tween-20s) and sterilize 10 minutes, with aseptic water washing 4 times, cut limb standby;
B, explant induction: will be through fritter standby in steps A, be seeded on callus inducing medium, this medium is: MS+6-BA 2.0 mg/L+2,4-D 0.2 mg/L+NAA 1.0 mg/L, add 1.0% agar, 3% sucrose, 0.5% active carbon and natural organic additive in medium.Incubation time is 45 days, in the incision of blade, expands and forms jade-green callus.
Described superclean bench, model is HS-1300, by SuZhou Antai Air Tech Co., Ltd. of Su Jing group, is produced;
C, indefinite bud induce differentiation: the callus obtained in step B is inoculated on the medium of adventitious bud inducing, this medium is: MS+6-BA 3.0 mg/L+NAA 0.5mg/L adds 1.0% agar, 3% sucrose, 0.5% active carbon and natural organic additive in medium.Cultivate 30 days, crowd shoots occurs;
D, culture of rootage: by the crowd shoots plant division obtained in step C, proceed in root media, this medium is: MS+NAA 0.05 mg/L adds 1.0% agar, 3% sucrose and 0.5% active carbon in medium, cultivate 30 days, the seedling base portion short of white occurs and grows complete root system;
E, hardening and transplanting: will obtain the bottle seedling of complete plantlet in step D, be put in the inside, greenhouse hardening 5 days, the bottle seedling is opened to hardening 2 days, then from bottle, remove plant, carefully wash away the agar medium adhered to, carry out at ambient temperature the Nutrition Soil cultivation.
Claims (8)
1. a tissue culture rapid speed is bred the method for safflower frangipanis, it is characterized in that: take MS as minimal medium, safflower frangipanis blade is that explant carries out tissue-culturing rapid propagation, after sterile-processed, inoculation is induced and is formed callus, again through Calli Differentiation, bud propagation and culture of rootage, form the whole plant seedling, finally carry out hardening and transplanting.
2. tissue culture rapid speed according to claim 1 is bred the method for safflower frangipanis, it is characterized in that: tissue culture rapid speed is bred the method for safflower frangipanis, comprises the following steps:
The selection of explant and disinfecting: after choosing the blade on the safflower frangipanis maternal plant of well-grown stalwartness, be put in temporarily the place two days in indoor cool place, after the juice discharged in blade is dry, after cleaning 10 minutes with running water, on aseptic super-clean bench, dry, then, blade is cut into to each fritter of 2 centimetres of length and width, fritter is put into to 75% alcohol rinsing once (in 10 seconds), proceed in 0.1% mercuric chloride solution (being added with five or six of Tween-20s) and sterilize 10 minutes, with aseptic water washing 3 ~ 4 times, cut limb standby;
Explant induction: will be seeded on callus inducing medium through fritter standby in steps A, incubation time is 40 ~ 50 days, in the incision of blade, expands and forms jade-green callus.
Indefinite bud induce differentiation: the callus obtained in step B is inoculated on the medium of adventitious bud inducing, cultivates 32 ~ 35 days, crowd shoots occurs;
Culture of rootage: the crowd shoots plant division by obtaining in step C, proceed in root media, to cultivate 30 ~ 33 days, the seedling base portion short of white occurs and grows complete root system;
Hardening and transplanting: will obtain the bottle seedling of complete plantlet in step D, be put in the inside, greenhouse hardening 5 days, the bottle seedling is opened to hardening 2 days, then from bottle, remove plant, carefully wash away the agar medium adhered to, carry out at ambient temperature the Nutrition Soil cultivation.
4. tissue culture rapid speed according to claim 2 is bred the method for safflower frangipanis, it is characterized in that: described medium of inducing with Callus of Leaf is MS, 6-BA 0 ~ 3 mg/L, 2.4-D 0 ~ 0.5mg/L, NAA 0 ~ 1mg/L, wherein: MS is conventional minimal medium Murashige&Skoog 1962,6-BA is 6-benzyl aminoadenine, 2,4-D is 2,4-dichlorphenoxyacetic acid, and NAA is methyl α-naphthyl acetate.
5. tissue culture rapid speed according to claim 2 is bred the method for safflower frangipanis, it is characterized in that: described adventitious bud inducing differential medium is: MS, 6-BA 1 ~ 3 mg/L, NAA 0.1 ~ 0.5 mg/L, wherein: MS is conventional minimal medium Murashige&Skoog 1962,6-BA is 6-benzyl aminoadenine, and NAA is methyl α-naphthyl acetate.
6. tissue culture rapid speed according to claim 2 is bred the method for safflower frangipanis, it is characterized in that: described root media is: the sucrose of 1/2MS, NAA 0.01 ~ 0.1mg/L, 2-3%, 0.8 ~ 1.0% agar, wherein: MS is conventional minimal medium Murashige&Skoog 1962, macroelement in this routine minimal medium is reduced by half and obtains 1/2MS, and NAA is methyl α-naphthyl acetate.
7. tissue culture rapid speed according to claim 2 is bred the method for safflower frangipanis, it is characterized in that: described explant induction medium and adventitious bud inducing differential medium all are incorporated as 1.0% agar, 2 ~ 3% sucrose, 0.5% active carbon and natural organic additive.
8. tissue culture rapid speed according to claim 1 and 2 is bred the method for safflower frangipanis, it is characterized in that: described natural organic additive is 0.2% coconut milk, 0.1% apple juice, 0.1% banana puree mixture.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103858768A (en) * | 2014-04-01 | 2014-06-18 | 中国热带农业科学院海口实验站 | Tissue culture method of plumeria rubra L.cv.Acutifolia |
| CN107494268A (en) * | 2017-09-29 | 2017-12-22 | 江苏农林职业技术学院 | The tissue culture and rapid propagation method of multicolored trachelospermum jasminoide |
| CN108812314A (en) * | 2018-06-22 | 2018-11-16 | 东北林业大学 | Ground fruit tissue cultures and Regeneration System |
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| WO2005012507A1 (en) * | 2003-07-25 | 2005-02-10 | The University Of Melbourne | Production of plant secondary metabolites using adsorption and elicitation in cell suspension culture |
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| WO2005012507A1 (en) * | 2003-07-25 | 2005-02-10 | The University Of Melbourne | Production of plant secondary metabolites using adsorption and elicitation in cell suspension culture |
| CN101406157A (en) * | 2008-11-27 | 2009-04-15 | 江苏农林职业技术学院绿苑实业总公司 | Tissue culture method of Nerium indicum |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103858768A (en) * | 2014-04-01 | 2014-06-18 | 中国热带农业科学院海口实验站 | Tissue culture method of plumeria rubra L.cv.Acutifolia |
| CN107494268A (en) * | 2017-09-29 | 2017-12-22 | 江苏农林职业技术学院 | The tissue culture and rapid propagation method of multicolored trachelospermum jasminoide |
| CN108812314A (en) * | 2018-06-22 | 2018-11-16 | 东北林业大学 | Ground fruit tissue cultures and Regeneration System |
| CN108812314B (en) * | 2018-06-22 | 2021-05-18 | 东北林业大学 | Establishment of ficus tikoua tissue culture and regeneration system |
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Application publication date: 20131218 |