CN106900550A - A kind of abductive approach of water shield Multiple Buds - Google Patents
A kind of abductive approach of water shield Multiple Buds Download PDFInfo
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- CN106900550A CN106900550A CN201710109598.1A CN201710109598A CN106900550A CN 106900550 A CN106900550 A CN 106900550A CN 201710109598 A CN201710109598 A CN 201710109598A CN 106900550 A CN106900550 A CN 106900550A
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- water
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
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- Botany (AREA)
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- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a kind of abductive approach of water shield Multiple Buds, belong to the field of tissue culture of water shield.Abductive approach step includes：Explant is selected and pretreatment, explant sterilizing, bud induction, bud Multiplying culture.Culture medium constituent is including a great number of elements, trace element, molysite and complexing agent, organic principle, inorganic additive, plant growth regulator, physiological activator, carbon source, coagulator and other additives etc..The invention provides the culture medium and cultural method of water shield inducing clumping bud, water shield bud induction rate can be effectively improved, and inductivity is high, the later stage can in a short time form a large amount of excellent test tube seedlings, cultivated for the extraordinary aquatic vegetable of China and provide seedling.The present invention can be not only used for the breeding of water shield, can also be applied to the fields such as water shield rapid propagation in vitro, preserving seed.
Description
Technical field
The present invention provides a kind of abductive approach of water shield Multiple Buds, belongs to field of plant tissue culture technique.
Background technology
Water shield, belongs to a kind of pasture and water of Nymphaeceae, lists I grade of Top-rated protected wild plants of China national in（State Council 1999 8
The moon 4 was ratified）, it is precious wild water vegetables, contain acidic polysaccharose, protein, amino acid, vitamin, histamine and micro
Element etc., its value is mainly reflected in medical value and edibility.Water shield contains abundant colloid albumen, carbohydrate,
Fat, multivitamin and mineral matter, often to eat water shield have the health-care effect of medicine-food two-purpose, are one of precious aquatic vegetables.
But due to not doing effective variety rejuvenation and kind protection for many years, water shield kind sexual involution has compared sternly
Weight, quality and yield are all declining.
Still lack mature technology at present, can be with vast propagation water shield high grade project seedling, while overcoming the species in nature
The problem that introduces a collection is limited in environment and kind is gradually degenerated.
The content of the invention
The invention provides a kind of culture medium and abductive approach of water shield inducing clumping bud, the method is in aseptic condition
Under, temperature control measure is taken, fixed-illumination is given, water shield Multiple Buds are induced, progressively achievement aseptic seedling system, the later stage can provide throughout the year
A large amount of aseptic seedlings.Easy to implement the method, easy to operate, yield is high.
In order to achieve the above object, the present invention uses following technical measures：
A kind of abductive approach of water shield Multiple Buds, comprises the following steps：
A, explant selection and pretreatment：Selection green in color, mud cover the water shield stem section of few axillary bud sprouted as outer
Implant, 10min is soaked with alkaline detergent, and surface mud is gently brushed off with banister brush to stem section in green, and 1 is rinsed with running water
Hour, rinsed 4-5 times repeatedly with distilled water, cleaned 4-5 times repeatedly again with sterilized water, it is stand-by；
B, explant sterilizing：Soaked 30 seconds with 75% ethanol first on aseptic operating platform, with aseptic water washing 4-5 times, then used
0.1% mercury chloride (adding a few drop Tween 80s) carries out surface sterilization 10 minutes, aseptic water washing 4-5 times, by water on aseptic filter paper
Blot, it is stand-by；
C, bud induction：The MS solid mediums of 1.0 mg/L 6-BA and 0.15 mg/L NAA are selected, sucrose mass concentration is 3%,
Quality of activated carbon concentration is adjusted to 5.8-6.0 for 0.4%, pH, and solid medium is loaded in tissue culture bottle；Solid medium passes through
120 DEG C of sterilization treatment 20min, after cooling, access the water shield explant of the 1-1.5cm after sterilizing, add sterilized water to exceed solid
Culture basal plane 3-4cm, is placed in illumination box and cultivates, and treating that young shoot is long carries out Multiplying culture to 2.0-3.0cm；
D, bud Multiplying culture：Selection sucrose mass concentration 3%, quality of activated carbon concentration is 0.4% MS solid mediums, addition
0.5-2.00 mg/L 6-BA, 2-4 mg/L 2,4-D, the hormone of 0.1-0.50 mg/L NAA；Culture medium packing, culture medium go out
Bacterium mode is identical with bud induction, adds sterilized water to exceed solid culture basal plane 3-4cm, accesses the young shoot of 2.0-3.0cm long in light
According to incubator, Multiplying culture is carried out；
Described step C, D is gnotobasis, illumination 60-70 μ E/（m2·s）, periodicity of illumination is 12h/d, indoor temperature（25
±2）℃.
Preferably, in step B, 0.1% mercury chloride is used（Add a few drop Tween 80s）Surface sterilization 10min is carried out, in step C,
The concentration of 6-BA is 1.0mg/L, and the concentration of NAA is 0.15mg/L.
The invention has the advantages that：
The current technology on water shield tissue cultures has no report, also and systematic research is not made to water shield.The technology can be straight
The growth of induction water shield young shoot is connect, Multiple Buds is constantly produced, for the foundation of water shield sterile propagation system provides research of technique.Water shield
Can transplant throughout the year, especially with transplanting survival rate highest before green sprouting, can harvest then again, can not be received using the present invention
Season, temperature, regional impact, obtain substantial amounts of Multiple Buds, and the later stage carries out culture of rootage etc., can quickly set up the aseptic of water shield again
Breeding system, for the kind for keeping water shield excellent provides technical support, can provide continuously aseptic for vast raiser
Seedling.One can reach 8 times for the explant of 1.5cm long by the culture bud growth coefficient of month.With a bud 30d
Once, each bud growth coefficient is 8 calculating to subculture, it is contemplated that result is as shown in table 1 below.
Table 1
| Number of days/d | 30 | 90 | 150 | 210 | 270 | 330 | 360 |
| Seedling number/ | 8 |
Brief description of the drawings
Fig. 1 is water shield 30d proliferative conditions in embodiment one,
Fig. 2 is water shield 30d proliferative conditions in embodiment two.
Specific embodiment
The reagent that the embodiment of the present invention is used, if not otherwise specified, commercially available is capable of achieving the present invention, involved skill
Art scheme, if not otherwise specified, can use the ordinary skill in the art.
The MS culture medium prescriptions used in embodiment are as follows：Unit mg/L
KNO3 1900
NH4NO3 1650
MgSO4•7H2O 370
KH2PO4 170
CaCl2•2H2O 440
MnSO4•4H2O 22.3
ZnSO4•7H2O 8.6
H3BO3 6.2
KI 0.83
Na2MoO4•7H2O 0.25
CuSO4.5H2O 0.025
CoCL2.6H2O 0.025
Na2-EDTA 37.3
FeSO4•7 H2O 27.8
Glycine 2.0
Puridoxine hydrochloride 0.5
Tyiamine Hd element 0.1
Nicotinic acid 0.5
Creatine 100.
MS solid mediums are addition 7g/L agar in above-mentioned culture formula.
Embodiment one
A, explant selection and pretreatment：Selection green in color, mud cover the water shield stem section of few axillary bud sprouted as outer
Implant, 10min is soaked with alkaline detergent, and surface mud is gently brushed off with banister brush to stem section in green, and 1 is rinsed with running water
Hour, rinsed 4-5 times repeatedly with distilled water, cleaned 4-5 times repeatedly again with sterilized water, it is stand-by；
B, explant sterilizing：Soaked 30 seconds with 75% ethanol first on aseptic operating platform, with aseptic water washing 4-5 times, then used
0.1% mercury chloride (adding a few drop Tween 80s) carries out surface sterilization 10 minutes, aseptic water washing 4-5 times, by water on aseptic filter paper
Blot, it is stand-by；
C, bud induction：The MS solid mediums of 1.0 mg/L 6-BA and 0.15 mg/L NAA are selected, sucrose mass concentration is 3%,
Quality of activated carbon concentration is adjusted to 5.8-6.0 for 0.4%, pH, and solid medium is loaded in tissue culture bottle；Solid medium passes through
120 DEG C of sterilization treatment 20min, after cooling, access the water shield explant of the 1-1.5cm after sterilizing, add sterilized water to exceed solid
Culture basal plane 3-4cm, is placed in illumination box and cultivates, and treating that young shoot is long carries out Multiplying culture to 2.0-3.0cm；
D, bud Multiplying culture：Selection sucrose mass concentration 3%, quality of activated carbon concentration is 0.4% MS solid mediums, addition
0.5mg/L 6-BA, 4 mg/L 2,4-D, the hormone of 0.1 mg/L NAA；Culture medium packing, medium sterilization mode and bud are lured
Lead identical, add sterilized water to exceed solid culture basal plane 3-4cm, access the young shoot of 2.0-3.0cm long in illumination box, lure
Lead a large amount of Multiple Buds；
Described step C, D, use gnotobasis, illumination 60-70 μ E/（m2·s）, periodicity of illumination is 12h/d, indoor temperature
（25±2）℃.
This method can directly induce young shoot by an explant, and breeding of or else breaking obtains more clump buds, 30 d's
Growth coefficient reaches 8, and survival rate is more than 90%.Fig. 1 is water shield 30d proliferative conditions in embodiment one.
Embodiment two
A, explant selection and pretreatment：Selection green in color, mud cover the water shield stem section of few axillary bud sprouted as outer
Implant, 10min is soaked with alkaline detergent, and surface mud is gently brushed off with banister brush to stem section in green, and 1 is rinsed with running water
Hour, rinsed 4-5 times repeatedly with distilled water, cleaned 4-5 times repeatedly again with sterilized water, it is stand-by；
B, explant sterilizing：Soaked 30 seconds with 75% ethanol first on aseptic operating platform, with aseptic water washing 4-5 times, then used
0.1% mercury chloride (adding a few drop Tween 80s) carries out surface sterilization 10 minutes, aseptic water washing 4-5 times, by water on aseptic filter paper
Blot, it is stand-by；
C, bud induction：The MS solid mediums of 1.0 mg/L 6-BA and 0.15 mg/L NAA are selected, sucrose mass concentration is 3%,
Quality of activated carbon concentration is adjusted to 5.8-6.0 for 0.4%, pH, and solid medium is loaded in tissue culture bottle；Solid medium passes through
120 DEG C of sterilization treatment 20min, after cooling, access the water shield explant of the 1-1.5cm after sterilizing, add sterilized water to exceed solid
Culture basal plane 3-4cm, is placed in illumination box and cultivates, and treating that young shoot is long carries out Multiplying culture to 2.0-3.0cm；
D, bud Multiplying culture：Selection sucrose mass concentration 3%, quality of activated carbon concentration is 0.4% MS solid mediums, addition
2.00 mg/L 6-BA, 2 mg/L 2,4-D, the hormone of 0.50 mg/L NAA；Culture medium packing, medium sterilization mode and bud
Induction is identical, adds sterilized water to exceed solid culture basal plane 3-4cm, accesses the young shoot of 2.0-3.0cm long in illumination box,
Carry out Multiplying culture；
Described step C, D uses gnotobasis, illumination 60-70 μ E/（m2·s）, periodicity of illumination is 12h/d, indoor temperature
（25±2）℃.
This method directly induces seedling by an explant, and the growth coefficient of 30 d can reach 6 times, and survival rate is more than
85%.Fig. 2 is water shield 30d proliferative conditions in embodiment two.
The present invention uses modern biotechnology means, and tissue cultures induction Multiple Buds are carried out to water shield, progressively sets up quick
Breeding sterile system, enables the rapid popularizing planting of improved seeds, and low production cost, cultivation cycle is short, realizes factorial praluction
Purpose, has broad application prospects and promotional value higher, remarkable in economical benefits.
Claims (3)
1. a kind of abductive approach of water shield Multiple Buds, it is characterised in that comprise the following steps：
A, explant selection and pretreatment：Selection green in color, mud covers the water shield stem section of few axillary bud sprouted as outer
Implant, 10min is soaked with alkaline detergent, removes surface mud, then is rinsed 1 hour with running water, is rinsed repeatedly with distilled water
4-5 times, cleaned 4-5 times repeatedly again with sterilized water, be placed in aseptic operating platform；
B, explant sterilizing：Soaked 30 seconds with 75% ethanol first on aseptic operating platform, with aseptic water washing 4-5 times, then carried out
Surface sterilization, aseptic water washing 4-5 times blots water on aseptic filter paper, stand-by；
C, bud induction：Select 1.0 mg/L 6-BA（6-benzyl aminopurine）With 0.15 mg/L NAA（Methyl α-naphthyl acetate）MS solids training
Base is supported, sucrose mass concentration is 3%, and quality of activated carbon concentration is adjusted to 5.8-6.0 for 0.4%, pH, and solid medium is loaded on tissue culture bottle
In；Solid medium after cooling, accesses the water shield explant of the 1-1.5cm after sterilizing by 120 DEG C of sterilization treatment 20min, plus
Enter sterilized water more than solid culture basal plane 3-4cm, be placed in illumination box and cultivate, treat that young shoot is long and increased to 2.0-3.0cm
Grow culture；
D, bud Multiplying culture：Selection sucrose mass concentration 3%, quality of activated carbon concentration is 0.4% MS solid mediums, addition
0.5-2.0 mg/L 6-BA、2-4 mg/L 2,4-D（2,4 dichlorophenoxyacetic acid）, 0.1-0.5 mg/L NAA hormone；Training
Foster base packing, medium sterilization mode are identical with bud induction, add sterilized water to exceed solid culture basal plane 3-4cm, access and grow
The young shoot of 2.0-3.0cm carries out Multiplying culture in illumination box；
Described step C, D uses gnotobasis, illumination 60-70 μ E/（m2·s）, periodicity of illumination is 12h/d, indoor temperature 25
±2℃。
2. method according to claim 1, it is characterised in that preferred, in step B, described surface sterilization is used
0.1% mercury chloride（Add a few drop Tween 80s）Carry out surface sterilization 10min.
3. method according to claim 1, it is characterised in that preferred, in step C, the growth regulating of culture medium addition
The concentration of material 6-BA is 1.0mg/L, and the concentration of NAA is 0.15mg/L.
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| CN201710109598.1A CN106900550B (en) | 2017-02-27 | 2017-02-27 | A kind of abductive approach of water shield Multiple Buds |
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| CN106900550B CN106900550B (en) | 2019-04-05 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109122311A (en) * | 2018-07-31 | 2019-01-04 | 海南大学 | A kind of culture medium and its cultural method of water lily viviparity stem tuber induction sterile bud |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102283122A (en) * | 2011-07-11 | 2011-12-21 | 镇江瑞繁农艺有限公司 | Dormancy breaking seeding raising method for water lily seeds |
| CN103461134A (en) * | 2013-09-22 | 2013-12-25 | 镇江瑞繁农艺有限公司 | Method for cultivating water lily aseptic seedling |
| CN104488716A (en) * | 2014-12-19 | 2015-04-08 | 浙江省农业科学院 | Method for tissue culture of water nymph |
| CN104996298A (en) * | 2015-07-06 | 2015-10-28 | 三明学院 | Method for cultivating polygala fallax hemsl tissue culture seedlings based on multiple internodal stem segments integration |
-
2017
- 2017-02-27 CN CN201710109598.1A patent/CN106900550B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102283122A (en) * | 2011-07-11 | 2011-12-21 | 镇江瑞繁农艺有限公司 | Dormancy breaking seeding raising method for water lily seeds |
| CN103461134A (en) * | 2013-09-22 | 2013-12-25 | 镇江瑞繁农艺有限公司 | Method for cultivating water lily aseptic seedling |
| CN104488716A (en) * | 2014-12-19 | 2015-04-08 | 浙江省农业科学院 | Method for tissue culture of water nymph |
| CN104996298A (en) * | 2015-07-06 | 2015-10-28 | 三明学院 | Method for cultivating polygala fallax hemsl tissue culture seedlings based on multiple internodal stem segments integration |
Non-Patent Citations (2)
| Title |
|---|
| 王蒂等: "《植物组织培养》", 31 August 2013 * |
| 黄国振等: "睡莲属植物的繁殖形式", 《中国花卉园艺》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109122311A (en) * | 2018-07-31 | 2019-01-04 | 海南大学 | A kind of culture medium and its cultural method of water lily viviparity stem tuber induction sterile bud |
| 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 |
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