CN101120655B - Method for making potamogeton cripus artificial seed - Google Patents
Method for making potamogeton cripus artificial seed Download PDFInfo
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- CN101120655B CN101120655B CN2007100532499A CN200710053249A CN101120655B CN 101120655 B CN101120655 B CN 101120655B CN 2007100532499 A CN2007100532499 A CN 2007100532499A CN 200710053249 A CN200710053249 A CN 200710053249A CN 101120655 B CN101120655 B CN 101120655B
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Abstract
The present invention discloses a production method for the artificial seed of the crispus, the production procedures of which is: firstly performing the introduced gemmulation on a solid cultivating medium containing the hormone; when the sprout is grown in a certain length, the sprout is adequately mixed with an embedding solution made with the sodium alginate as the embedding agent; and then drop the embedding solution containing the sprout with a dropper into the calcium chloride to perform the ion exchange reaction, and finally the sprout is rinsed with the sterile water to be made into the artificial seed of the crispus. The present invention resolves the problems the seed seedling source of the submerged plants in the rebuilding process of the water ecological system, which avoids damaging the plant resources at the seed-source place; moreover, the materials are easy to acquire, the process flowing procedures are simple, the present invention is not influenced by the season, the breeding speed is fast, the survival rate is high and the cost is low.
Description
Technical field
The invention belongs to the artificial plant seed production technical field, the key technology of potamogeton cripus artificial seed making mainly is provided, be specifically related to the propagation of inducing of water caltrop seedling bud, the preparation of potamogeton cripus artificial seed embedding liquid.The preparation method that more specifically relates to a kind of potamogeton cripus artificial seed is applicable to aquatic ecosystem reconstruction engineering and the landscape water body engineering demand to the submerged plant seedling.
Background technology
Artificial seed (artificial seeds) is meant that meristematic tissue (bud, callus, embryoid etc.) that the somatic embryo that will produce in the plant cultured in vitro maybe can develop into whole plant is embedded in and forms in the shell that contains nutriment and have defencive function, granule that under optimum conditions can germination and emergence.Artificial seed belongs to vegetative propagation in itself, compares with natural seed, and it is fast, with short production cycle that it has reproduction speed, but batch production mass preparation, storage, promotes rapidly, produces and advantage such as be not subject to seasonal restrictions.Up to now, the research range of artificial seed mainly concentrates on some important farm-forestry crops, medicinal plant and ornamental plants etc.
Submerged plant is the main member of Freshwater ecosystems, not only plays an important role aspect the aquatic ecosystem 26S Proteasome Structure and Function keeping, and also has the function of holding limpid water quality, can also increase the attractive in appearance of landscape water body simultaneously.
The recovery of submerged plant becomes the key link that water pollutes the comprehensive regulation.Because the submerged plant seed is difficult to obtain mostly, when the plant species word bank of water body was subjected to heavy damage, the recovery engineering of aquatic vegetation unavoidably had wilderness demand to the water plants seedling.The seedling of the engineering of aquatic vegetation recovery at present all is to fish for from natural water, and this has just caused destruction to the plant population of provenance.In addition, the acquisition of seedling also can be subjected to limitting season, and is subjected to the transportation range restriction.Utilize the artificial seed technology, can avoid destruction, and operate simple and easyly, not limited by season and transportation range the provenance plant resources.
Although quick propagating technology makes that many plants that are difficult to breed are bred at present, but and year-round provision test-tube plantlet.But property demand season of water plants seedling is very strong, because the restriction of culturing room space and manpower generally is difficult in a large amount of test-tube plantlets of centralizedly supply in a short time; Though present also someone studies the problems such as storage of test-tube plantlet, but storage in bulk and transportation are quite difficult, the restriction of more power consumption and manpower and materials when being subjected to culturing room space, low tempertaure storage, and still will be when needing these test-tube plantlets of plantation through the process of bottle outlet and transplanting, this process is followed or high or low death rate inevitably, and consumes more manpower and materials in the hardening process.Thereby make that the test-tube plantlet cost is too high.Artificial seed can conveniently store, and takes up an area of for a short time, and convenient transportation can remedy this type of deficiency.
Water caltrop (Potamogeton crispus L.) is the common submerged plant of China, and durability against pollution and dirty removal capacities are preferably arranged.Up to now, by retrieval, do not see the report of potamogeton cripus artificial seed as yet.
Summary of the invention
The objective of the invention is to be to provide a kind of preparation method of potamogeton cripus artificial seed, effectively solve the source problem that comes that aquatic vegetation recovers water caltrop seed and seedling in the engineering, material is easy to obtain, technological process simply, is not subjected to seasonal effect, reproduction speed is fast, survival rate is high, with low cost.
In order to achieve the above object, the present invention has adopted following technical scheme:
1, inducing of water caltrop bud: get aseptic water caltrop seedling, cut the stem section of the band joint of 4-6mm with scalpel,, be inoculated into to induce in the medium of sprouting and cultivate as group training germination explant, its composition is: MS or 1/5MS or 1/10MS+1.5-3% sucrose+0.8% agar+hormone, specific as follows:
(1) MS (Murashige and Skoog) composition:
Macroelement: KNO
3Be 1900mg/L, NH
4NO
3Be 1650mg/L, MgSO
4.7H
2O is 370mg/L, KH
2PO
4Be 170mg/L, CaCl
2.2H
2O is 440mg/L;
Trace element: MnSO
4.4H
2O is 22.3mg/L, ZnSO
4.7H
2O is 8.6mg/L, H
3BO
3Be 6.2mg/L, KI is 0.83mg/L, NaMoO
4.2H
2O is 0.25mg/L, CuSO
4.5H
2O is 0.025mg/L, CoCl
2.6H
2O is 0.025mg/L;
Molysite: Na
2-EDTA is 37.3mg/L, FeSO
4.4H
2O is 27.8mg/L;
Organic principle: glycine is 2.0mg/L, and thiamine hydrochloride is 0.1mg/L, and puridoxine hydrochloride is 0.5mg/L for 0.5mg/L nicotinic acid, and inositol is 100mg/L;
(2) contained hormone kind: 6-BA, IBA, NAA
Induce the hormone combination of sprouting in the medium:
6-BA 1.0-5.0mgL
-1+ IBA 0.5mgL
-1Or NAA 0.5mgL
-1
(3) medium proportioning mode is:
MS+1.5-3% sucrose+0.8% agar+hormone or
MS/5+1.5-3% sucrose+0.8% agar+hormone or
MS/10+1.5-3% sucrose+0.8% agar+hormone or
Medium pH is 5.8-6.0.
2, after 4-6 days, eustipes part grows sprouting.The bud of the long 3-5mm that induces placed with tweezers contain 2%-5% sodium alginate+MS or 1/5MS or 1/10MS+1.5-3% sucrose+IBA 1.0mgL
-1+ 6-BA0.5mgL
-1Compound embedding liquid in, stir with glass bar bud and embedding liquid fully mixed;
3, after 10-15 minute, in superclean bench, the embedding drop that will contain bud with the dropper of internal diameter 3-5mm is gone into the calcium chloride (CaCl of 1-4%
2) in, promptly form the artificial seed of the similar capsule ball of white, translucent and tool certain degree of hardness through 15-30 minute ion-exchange reactions;
4, promptly get potamogeton cripus artificial seed with rinsed with sterile water 4-5 time with the termination ion-exchange reactions.In low temperature 3-5 ℃ of following lucifuge stored seeds.
The present invention compared with prior art, have the following advantages and effect: 1. can realize the large-scale breeding fast of water caltrop, avoided in the hydrophyte restoration engineering when a large amount of seedling of needs by fishing for from natural water, and the destruction that the plant population of provenance is caused; 2. produce and be not subjected to the restriction in season, fish for plant and then be subjected to the influence in season bigger from natural water, the kind shoot survival percent that different time is gathered is also different; 3. material source is easy, operates simple and easyly, 4. compares with test-tube plantlet, and volume is little, transportation and preserve convenient, used cultivation base unit weight is few, with low cost, has reduced transplanting domestication process.
Description of drawings
Fig. 1: the potamogeton cripus artificial seed that is made into, outside are white, translucent, people's work post skin of similar capsule ball, and inside is the bud of the water caltrop that induces;
Fig. 2: potamogeton cripus artificial seed is sprouted in unpasteurized distilled water and is changeed strain, and artificial seed promptly began to sprout in after planting 3-5 days, and growth rate is very fast, and green is the cauline leaf of water caltrop plant among the figure, and white, elongated is the root of water caltrop plant.
Embodiment
The present invention is a kind of preparation method of potamogeton cripus artificial seed, specifically implements as follows:
Embodiment 1:
1, inducing of bud: the experiment outfit needs through 121 ℃, 0.1MPa high pressure steam sterilization as flat board, scalpel, tweezers and medium etc., before experiment, above experimental tool will be positioned in the superclean bench and sterilize 30 minutes, remove Ye Hegen with scalpel and tweezers then, cut the stem section of the band joint of 4-6mm, as group training germination explant, be inoculated into to induce in the medium of sprouting and cultivate, its composition is: MS or 1/5MS or 1/10MS+1.5% or 1.8% or 2.1% or 2.4% or 2.7% or 3% sucrose+0.8% agar+hormone, specific as follows:
(1) MS (Murashige and Skoog) composition:
Macroelement: KNO
3Be 1900mg/L, NH
4NO
3Be 1650mg/L, MgSO
4.7H
2O is 370mg/L, KH
2PO
4Be 170mg/L, CaCl
2.2H
2O is 440mg/L;
Trace element: MnSO
4.4H
2O is 22.3mg/L, ZnSO
4.7H
2O is 8.6mg/L, H
3BO
3Be 6.2mg/L, KI is 0.83mg/L, NaMoO
4.2H
2O is 0.25mg/L, CuSO
4.5H
2O is 0.025mg/L, CoCl
2.6H
2O is 0.025mg/L;
Molysite: Na
2-EDTA is 37.3mg/L, FeSO
4.4H
2O is 27.8mg/L;
Organic principle: glycine is 2.0mg/L, and thiamine hydrochloride is 0.1mg/L, and puridoxine hydrochloride is 0.5mg/L, and nicotinic acid is 0.5mg/L, and inositol is 100mg/L;
Described 1/5MS refers to that macroelement and trace element are 1/5 of MS medium full dose, and molysite and organic principle use the full dose of MS medium;
Described 1/10MS refers to that macroelement and trace element are 1/10 of MS medium full dose, and molysite and organic principle use the full dose of MS medium;
(2) contained hormone kind: 6-BA, IBA, NAA
Induce the hormone combination of sprouting to be in the medium:
6-BA 1.0-5.0mgL
-1+ IBA 0.5mgL
-1Or NAA 0.5mgL
-1
(3) medium proportioning mode is:
MS+1.5-3% sucrose+0.8% agar+hormone or
MS/5+1.5-3% sucrose+0.8% agar+hormone or
MS/10+1.5-3% sucrose+0.8% agar+hormone or
Medium pH is 5.8-6.0.
2, about after 4-6 days, eustipes part grows sprouting.The bud of the long 3-5mm that will induce in superclean bench places with tweezers and contains 2% or 2.5% or 3% or 3.5% or 4% or 4.5% or 5% sodium alginate+MS or 1/5MS or 1/10MS+1.5% or 1.8% or 2.1% or 2.4% or 2.7% or 3% sucrose+IBA 1.0mgL
-1+ 6-BA 0.5mgL
-1Embedding liquid in, stir with glass bar bud and embedding liquid fully mixed.
3, behind the 10-15min, in superclean bench, the embedding drop that will contain bud with the dropper of internal diameter 4mm is gone into 1% or 2% or 3% or 4% CaCl
2In, promptly form the artificial seed of the similar capsule ball of white, translucent and tool certain degree of hardness through the ion-exchange reactions of 20min.
4, promptly get potamogeton cripus artificial seed with rinsed with sterile water 4-5 time with the termination ion-exchange reactions.
Embodiment 2:
Induce the hormone combination of sprouting to be in the medium: or
6-BA 3.0mgL
-1+ IBA0.5mgL
-1Or NAA0.5mgL
-1
Implementation step is identical with embodiment 1.
Embodiment 3:
Induce the hormone combination of sprouting to be in the medium:
6-BA 5.0mgL
-1+ IBA0.5mgL
-1Or NAA0.5mgL
-1
Implementation step is identical with embodiment 1.
Embodiment 4:
Embedding medium sodium alginate concentration 2%, hormone combination:
IBA?1.0mg·L
-1+6-BA?0.5mg·L
-1。
Implementation step is identical with embodiment 1.
Embodiment 5:
Embedding medium sodium alginate concentration 5%, hormone combination:
IBA?1.0mg·L
-1+6-BA?0.5mg·L
-1。
Implementation step is identical with embodiment 1.
Embodiment 6:
After bud and embedding liquid fully mixed, in superclean bench, the embedding drop that will contain bud with the dropper of internal diameter 3mm was gone into 3% CaCl
2In, carry out the ion-exchange reactions of 30min.
Implementation step is identical with embodiment 1.
Embodiment 7:
After bud and embedding liquid fully mixed, in superclean bench, the embedding drop that will contain bud with the dropper of internal diameter 4mm was gone into 1% CaCl
2In, carry out the ion-exchange reactions of 20min.
Implementation step is identical with embodiment 1.
Embodiment 8:
After bud and embedding liquid fully mixed, in superclean bench, the embedding drop that will contain bud with the dropper of internal diameter 5mm was gone into 4% CaCl
2In, carry out the ion-exchange reactions of 15min.
Implementation step is identical with embodiment 1.
Embodiment 9:
The potamogeton cripus artificial seed of making is seeded in the distilled water, and sprouting condition is 20 ℃, intensity of illumination 36 μ molm
-2s
-1, light dark period is 12h:12h, the statistics germination rate is 70% after one month.
Implementation step is identical with embodiment 1.
Claims (1)
1. method for making potamogeton cripus artificial seed, it comprises the following steps:
Inducing of A, water caltrop bud: get aseptic water caltrop seedling, cut the stem section of the band joint of 4-6mm with cutter, as group training germination explant, be inoculated into to induce in the medium of sprouting and cultivate, its composition is: MS or 1/5MS or 1/10MS+1.5-3% sucrose+0.8% agar+hormone;
After B, 4-6 days, eustipes part grows sprouting, and the bud of the long 3-5mm that induces is placed embedding liquid with tweezers, stirs with glass bar bud and embedding liquid are mixed;
After C, 10-15 minute, in superclean bench, the embedding drop that will contain bud with the dropper of internal diameter 4mm is gone into 1% CaCl
2In, promptly form white, translucent artificial seed through 20 minutes ion-exchange reactions;
D, promptly get potamogeton cripus artificial seed with the termination ion-exchange reactions with rinsed with sterile water 4-5 time;
Described MS medium is:
Macroelement: KNO
3Be 1900mg/L, NH
4NO
3Be 1650mg/L, MgSO
4.7H
2O is 370mg/L, KH
2PO
4Be 170mg/L, CaCl
2.2H
2O is 440mg/L;
Trace element: MnSO
4.4H
2O is 22.3mg/L, ZnSO
4.7H
2O is 8.6mg/L, H
3BO
3Be 6.2mg/L, KI is 0.83mg/L, NaMoO
4.2H
2O is 0.25mg/L, CuSO
4.5H
2O is 0.025mg/L, CoCl
2.6H
2O is 0.025mg/L;
Molysite: Na
2-EDTA is 37.3mg/L, FeSO
4.4H
2O is 27.8mg/L;
Organic principle: glycine is 2.0mg/L, and thiamine hydrochloride is 0.1mg/L, and puridoxine hydrochloride is 0.5mg/L for 0.5mg/L nicotinic acid, and inositol is 100mg/L;
The hormone combination of inducing of described bud is:
6-BA?1.0-5.0mg·L
-1+IBA?0.5mg·L
-1；
Described embedding liquid compositing formula is:
2-5% sodium alginate+MS or 1/5MS or 1/10MS+1.5-3% sucrose+IBA 1.0mgL
-1+ 6-BA 0.5mgL
-1
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| CN101120655B true CN101120655B (en) | 2010-11-17 |
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| CN103477979B (en) * | 2013-09-06 | 2016-08-31 | 华南师范大学 | A kind of artificial seed preparation method of the black algae of submerged plant |
| CN103875527B (en) * | 2013-12-13 | 2016-04-13 | 岭南园林股份有限公司 | The artificial seed preparation method of a kind of submerged plant eel grass |
| CN103947547B (en) * | 2014-01-22 | 2016-07-13 | 华南师范大学 | A kind of improvement manufacture method of submerged plant artificial endosperm |
| CN103959958B (en) * | 2014-05-21 | 2016-03-30 | 四川农业大学 | A kind of preparation method of Hemarthria compressa artificial seed |
| CN104604681B (en) * | 2015-01-23 | 2017-12-01 | 福建省亚热带植物研究所 | A kind of method for improving plant tissue culture explant survival rate |
| CN104521762B (en) * | 2015-01-23 | 2017-02-22 | 福建省亚热带植物研究所 | Method for lightening microbiological contamination of plant tissue culture |
| CN104521763B (en) * | 2015-01-23 | 2017-03-15 | 福建省亚热带植物研究所 | A kind of method for mitigating plant tissue culture Brown |
| CN111316788A (en) * | 2020-02-27 | 2020-06-23 | 清上(苏州)环境科技有限公司 | Aquatic plant artificial seed and preparation method and application thereof |
| CN113317203A (en) * | 2021-06-28 | 2021-08-31 | 大连大学 | Pogostemon cablin artificial seed culture method |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1148462A (en) * | 1996-03-15 | 1997-04-30 | 中国科学院化学研究所 | Artificial seed and preparation method thereof |
| CN1742563A (en) * | 2005-10-13 | 2006-03-08 | 华中师范大学 | Method for fast breeding water caltrop seedlings |
| CN1943324A (en) * | 2005-10-13 | 2007-04-11 | 华中师范大学 | Method for fast breeding water caltrop seed and seedling |
-
2007
- 2007-09-18 CN CN2007100532499A patent/CN101120655B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1148462A (en) * | 1996-03-15 | 1997-04-30 | 中国科学院化学研究所 | Artificial seed and preparation method thereof |
| CN1742563A (en) * | 2005-10-13 | 2006-03-08 | 华中师范大学 | Method for fast breeding water caltrop seedlings |
| CN1943324A (en) * | 2005-10-13 | 2007-04-11 | 华中师范大学 | Method for fast breeding water caltrop seed and seedling |
Non-Patent Citations (4)
| Title |
|---|
| H Kunii.Life Cycle and Growth of Potamogeton crispus L.in a Shallow Pond, Ojaga-ike.《Journal of plant research》.1982,第95卷(第2期),109-124. * |
| HKunii.LifeCycleandGrowthofPotamogetoncrispusL.inaShallowPond Ojaga-ike.《Journal of plant research》.1982 |
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