CN104488716B - A kind of method of water lily tissue cultures - Google Patents
A kind of method of water lily tissue cultures Download PDFInfo
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- CN104488716B CN104488716B CN201410798205.9A CN201410798205A CN104488716B CN 104488716 B CN104488716 B CN 104488716B CN 201410798205 A CN201410798205 A CN 201410798205A CN 104488716 B CN104488716 B CN 104488716B
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Abstract
The invention discloses a kind of method of water lily tissue cultures, concrete operation step is as follows: the 1) preparation of medium, comprises the component of minimal medium and each stage medium of group training; 2) the choosing and sterilizing of explant; 3) induction of indefinite bud and propagation; 4) culture of rootage; 5) transplant.The effect that the present invention is useful: the present invention utilizes plant tissue culture technique to carry out adventitious bud inducing and Multiplying culture to water lily; the high-quality strong sprout that a large amount of genetic character is consistent can be obtained at short notice; overcome the shortcoming that Sterile culture method is slow, all there is positive effect to its large-scale production and Germ-plasma resources protection simultaneously.
Description
Technical field
The present invention relates to plant tissue culture technique, be specifically related to a kind of method of water lily tissue cultures.
Background technology
Water lily (NymphaeatetragonaGeorgi) is the perennial aquatic herbaceous plant of Nymphaeceae Nymphaea, and floral leaf is all beautiful, and pattern enriches, and flowering stage is long, and ornamental value is high; The root of water lily can absorb the noxious material such as lead, mercury, phenol in water, is rare water body purification plant; In addition water lily also has certain medical value.But water lily many dependences division propagation and seed propagation, reproduction coefficient is low, the cycle is long, is difficult to the needs meeting market.Therefore; plant tissue culture technique is utilized to carry out the Fast-propagation of water lily; a large amount of seedling consistent with maternal plant genetic character can obtained more in a short time; this for preservation fine germplasm resources, breed famous-brand and high-quality water lily kind and have great importance, also have practical using value for realizing its large-scale production with the demand meeting domestic and international market simultaneously.But, before this also not about the report of water lily Study on tissue culture, more there is no successful example.
Summary of the invention
The object of the invention is to the deficiency overcoming prior art existence, and a kind of method of water lily tissue cultures is provided.
For achieving the above object, the technical solution used in the present invention is: the method for this water lily tissue cultures mainly comprises the steps:
1), the preparation of medium:
(1) minimal medium: adopt double layer culture base, its upper strata is sterile distilled water; Lower floor is MS solid culture medium, wherein sucrose 20 ~ 30g/L, agar 5 ~ 9g/L, pH=5.8;
(2) inducing culture solid layer: MS+TDZ1.0 ~ 3.0mg/L+NAA0.05 ~ 0.5mg/L+ active carbon 0.2 ~ 0.5mg/L; (3) proliferated culture medium solid layer: MS+6-BA2.0 ~ 4.0mg/L+NAA0.1 ~ 0.5mg/L;
(4) root media solid layer: MS+NAA0.05 ~ 0.2mg/L+IBA0.05 ~ 0.2mg/L;
2) the choosing and sterilizing of explant: repair smooth by the terminal bud on water lily rhizome and lateral bud periphery, cleans and for subsequent use after sterilizing;
3) induction of indefinite bud and propagation: by step 2) disinfect after sprout be aseptically inoculated in the solid layer of inducing culture, directly can induce indefinite bud after 1 ~ 2 month, indefinite bud is transferred on proliferated culture medium and can must breed further;
4) culture of rootage: by step 3) propagation bud clump is transferred on root media after being aseptically divided into individual plant, and cultivate and can take root after 15 ~ 25 days;
5) transplant: by step 4) the water lily plant that takes root takes out and cleans, then transplanting and cultivate to muddy water groove from medium.
Further, the present invention is in described step 1) in, the component that described medium comprises minimal medium and each stage medium of group training is specially:
(1) minimal medium: adopt double layer culture base, its upper strata is sterile distilled water; Lower floor is MS solid culture medium, wherein sucrose 20 ~ 30g/L, agar 5 ~ 9g/L, pH=5.8;
(2) inducing culture solid layer: MS+TDZ2.0mg/L+NAA0.5mg/L+ active carbon 0.2mg/L;
(3) proliferated culture medium solid layer: MS+6-BA3.0mg/L+NAA0.2mg/L;
(4) strong seedling culture base solid layer: MS+NAA0.1mg/L+IBA0.1mg/L.
Further, the present invention is in described step 2) in, described disinfecting takes after water lily rhizome removes blade terminal bud, lateral bud to cut together with part rhizome tissue, with sharp blade the blade of sprout periphery and batting excised and repair smooth rear saturated washing powder solution soak in 20 ~ 60min, and then it is clean with tap water, then the alcohol, the effective chlorine density that are 70% in volume ratio be successively 1% liquor natrii hypochloritis and 0.1% mercuric chloride in soak 0.5 ~ 1min, 4 ~ 8min and 4 ~ 6min respectively, finally use aseptic water washing 3 ~ 5 times.
Further, the present invention is in described step 3) in, described in disinfect after sprout be directly inoculated in evoking adventive bud eruption in the solid layer of inducing culture, be transferred to after the indefinite bud induced is cut in proliferated culture medium carry out propagation expand numerous.
Further, the present invention is in described step 5) in, described transplanting is taken out from medium and clean root remaining medium by the water lily plant after taking root, and then transplants and cultivate to the cultivating groove that muddy water is housed.
Beneficial effect of the present invention is: utilize plant tissue culture technique to carry out cultured in vitro to water lily; the high-quality strong sprout that a large amount of genetic character is consistent can be obtained within a short period of time; overcome the shortcoming that Sterile culture method is slow, all there is positive effect to its large-scale production and Germ-plasma resources protection.
Embodiment
Below by embodiment, the present invention is further elaborated, and help is understood the present invention by embodiment better, but the present invention is not limited only to following embodiment.
The invention provides a kind of method of water lily tissue cultures, the steps include:
1), the preparation of medium
(1) minimal medium: adopt double layer culture base, its upper strata is sterile distilled water; Lower floor is MS solid culture medium, wherein sucrose 20 ~ 30g/L, agar 5 ~ 9g/L, pH=5.8;
(2) inducing culture solid layer: MS+TDZ1.0 ~ 3.0mg/L+NAA0.05 ~ 0.5mg/L+ active carbon 0.2 ~ 0.5mg/L;
(3) proliferated culture medium solid layer: MS+6-BA2.0 ~ 4.0mg/L+NAA0.1 ~ 0.5mg/L;
(4) root media solid layer: MS+NAA0.05 ~ 0.2mg/L+IBA0.05 ~ 0.2mg/L;
2), the choosing and sterilizing of explant
Take after water lily rhizome removes blade and terminal bud, lateral bud are cut together with part rhizome tissue, with sharp blade the blade of sprout periphery and batting excised and repair smooth rear saturated washing powder solution and soak 20 ~ 60min, and then it is clean with tap water, then the alcohol, the effective chlorine density that are 70% in volume ratio be successively 1% liquor natrii hypochloritis and 0.1% mercuric chloride in soak 0.5 ~ 1min, 4 ~ 8min and 4 ~ 6min respectively, finally use aseptic water washing 3 ~ 5 times;
3), the induction of indefinite bud and Multiplying culture
Sprout after disinfecting directly is inoculated on inducing culture, and cultivating after 1 ~ February can evoking adventive bud eruption, is transferred to that proliferated culture medium to carry out propagation expansion numerous after then being cut by the indefinite bud induced, and growth coefficient is 2 ~ 3; Cultivation temperature is 23 ± 2 DEG C, intensity of illumination is 80 ~ 120 μm of olm
-2s
-1, light application time is 8 ~ 12 hours/day;
4), culture of rootage
Be transferred on root media after propagation Multiple Buds is aseptically divided into individual plant, cultivate about 0.5cm of can taking root after 15 ~ 25 days; Cultivation temperature is 23 ± 2 DEG C, intensity of illumination is 40 ~ 80 μm of olm
-2s
-1, light application time is 8 ~ 12 hours/day;
5), transplant
Described transplanting is taken out from medium and clean root remaining medium by the water lily plant after taking root, and then transplants and cultivate to the cultivating groove that muddy water is housed; Transplanting survival rate about 85%.
Embodiment
The invention provides a kind of method of water lily tissue cultures, the steps include:
1), the preparation of medium
(1) minimal medium: adopt double layer culture base, its upper strata is sterile distilled water; Lower floor is MS solid culture medium, wherein sucrose 20 ~ 30g/L, agar 5 ~ 9g/L, pH=5.8;
(2) inducing culture solid layer: MS+TDZ2.0mg/L+NAA0.5mg/L+ active carbon 0.2mg/L;
(3) proliferated culture medium solid layer: MS+6-BA3.0mg/L+NAA0.2mg/L;
(4) root media solid layer: MS+NAA0.1mg/L+IBA0.1mg/L;
2), the choosing and sterilizing of explant
Take after water lily rhizome removes blade and terminal bud, lateral bud are cut together with part rhizome tissue, with sharp blade the blade of sprout periphery and batting excised and repair smooth rear saturated washing powder solution and soak 60min, and then it is clean with tap water, then the alcohol, the effective chlorine density that are 70% in volume ratio be successively 1% liquor natrii hypochloritis and 0.1% mercuric chloride in soak 1min, 5min and 5min respectively, finally use aseptic water washing 5 times;
3), the induction of indefinite bud and Multiplying culture
Sprout after disinfecting directly is inoculated on inducing culture, and cultivating after 1 ~ February can evoking adventive bud eruption, is transferred to that proliferated culture medium to carry out propagation expansion numerous after then being cut by the indefinite bud induced, and growth coefficient is about 3; Cultivation temperature is 23 ± 2 DEG C, intensity of illumination is 40 ~ 80 μm of olm
-2s
-1, light application time is 8 ~ 12 hours/day;
4), culture of rootage
Be transferred on root media after propagation Multiple Buds is aseptically divided into individual plant, cultivate about 0.8cm of can taking root after 15 ~ 25 days; Cultivation temperature is 23 ± 2 DEG C, intensity of illumination is 40 ~ 80 μm of olm
-2s
-1, light application time is 12 hours/day;
5), transplant
Described transplanting is taken out from medium and clean root remaining medium by the water lily plant after taking root, and then transplant and cultivate to the cultivating groove that muddy water is housed, transplanting survival rate can reach 90%.
Finally it should be noted that, except the present embodiment, the present invention can also have other embodiment and distortion, and what more than enumerate is only specific embodiments of the invention.All distortion that all those of ordinary skill in the art can directly derive from content disclosed by the invention or associate, all should think protection scope of the present invention.
Claims (4)
1. a method for water lily tissue cultures, is characterized in that: the method comprises the steps:
1), the preparation of medium:
(1) minimal medium: adopt double layer culture base, its upper strata is sterile distilled water; Lower floor is MS solid culture medium, wherein sucrose 20 ~ 30g/L, agar 5 ~ 9g/L, pH=5.8;
(2) inducing culture solid layer: MS+TDZ1.0 ~ 3.0mg/L+NAA0.05 ~ 0.5mg/L+ active carbon 0.2 ~ 0.5mg/L; (3) proliferated culture medium solid layer: MS+6-BA2.0 ~ 4.0mg/L+NAA0.1 ~ 0.5mg/L;
(4) root media solid layer: MS+NAA0.05 ~ 0.2mg/L+IBA0.05 ~ 0.2mg/L;
2) the choosing and sterilization of explant: repair smooth by the terminal bud on water lily rhizome and lateral bud periphery, cleans and for subsequent use after sterilizing;
3) induction of indefinite bud and propagation: by step 2) disinfect after bud be aseptically inoculated on inducing culture, directly can induce indefinite bud after 1 ~ 2 month, indefinite bud is transferred to the enterprising step of proliferated culture medium and breed;
4) culture of rootage: by step 3) propagation obtain indefinite bud aseptically cut after be transferred on root media, cultivate and take root after 15 ~ 25 days;
5) transplant: by step 4) take root after water lily plant take out from medium and clean, then transplant and cultivate to muddy water groove.
2. the method for water lily tissue cultures according to claim 1, is characterized in that: in described step 1) in, the component that described medium comprises minimal medium and each stage medium of group training is specially:
(1) minimal medium: adopt double layer culture base, its upper strata is sterile distilled water; Lower floor is MS solid culture medium, wherein sucrose 20 ~ 30g/L, agar 5 ~ 9g/L, pH=5.8;
(2) inducing culture solid layer: MS+TDZ2.0mg/L+NAA0.5mg/L+ active carbon 0.2mg/L;
(3) proliferated culture medium solid layer: MS+6-BA3.0mg/L+NAA0.2mg/L;
(4) root media solid layer: MS+NAA0.1mg/L+IBA0.1mg/L.
3. the method for water lily tissue cultures according to claim 1, it is characterized in that: in described step 2) in, described disinfecting is: take water lily rhizome and remove terminal bud after blade, lateral bud cuts together with part rhizome tissue, with sharp blade, smooth for the finishing of sprout periphery rear saturated washing powder solution is soaked 20 ~ 60min, and then it is clean with tap water, then be the alcohol of 70% at volume by volume concentration successively, effective chlorine density be 1% liquor natrii hypochloritis and 0.1% mercuric chloride solution in soak 0.5 ~ 1min respectively, 4 ~ 8min and 4 ~ 6min, finally use aseptic water washing 3 ~ 5 times.
4. the method for water lily tissue cultures according to claim 1, it is characterized in that: in described step 3) in, the induction of described indefinite bud and propagation are: the sprout after disinfecting is inoculated in evoking adventive bud eruption in the solid layer of inducing culture, be transferred in proliferated culture medium and breed after being cut by the indefinite bud induced.
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| CN106577272B (en) * | 2015-10-14 | 2019-07-05 | 西南大学 | A kind of method for tissue culture of water shield |
| CN106900550B (en) * | 2017-02-27 | 2019-04-05 | 水生藻安生物科技（武汉）有限公司 | A kind of abductive approach of water shield Multiple Buds |
| CN109122311B (en) * | 2018-07-31 | 2021-11-26 | 海南大学 | Culture medium for inducing aseptic buds of water lily embryonic tubers and culture method thereof |
| CN110839369B (en) * | 2019-11-15 | 2021-11-30 | 上海辰山植物园 | Preparation method of water lily sterile material |
| CN114303952B (en) * | 2021-12-31 | 2023-03-31 | 中国热带农业科学院海口实验站 | Water lily tuber induction tissue culture rapid propagation method |
| CN114467760B (en) * | 2022-04-15 | 2022-07-26 | 三亚南京农业大学研究院 | Water lily mature embryo callus induction and proliferation method |
| CN115644065A (en) * | 2022-12-01 | 2023-01-31 | 上海辰山植物园 | Rapid propagation method of fetal water lily |
Citations (1)
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| CN102283122B (en) * | 2011-07-11 | 2013-02-27 | 镇江瑞繁农艺有限公司 | Dormancy breaking seeding raising method for water lily seeds |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102283122B (en) * | 2011-07-11 | 2013-02-27 | 镇江瑞繁农艺有限公司 | Dormancy breaking seeding raising method for water lily seeds |
Non-Patent Citations (2)
| Title |
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| 睡莲不同杂交组合幼胚愈伤组织诱导研究;孙春青等;《江西农业学报》;20140131;第26卷(第1期);49-52 * |
| 碗莲组培快繁技术初探;孔德政等;《中国农学通报》;20080430;第24卷(第4期);76-79 * |
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