CN104705194A - Hormone-induced rapid propagation method of potamogeton pectinatus engineering seedlings - Google Patents
Hormone-induced rapid propagation method of potamogeton pectinatus engineering seedlings Download PDFInfo
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- 241000544047 Stuckenia pectinata Species 0.000 title claims abstract description 48
- 238000000034 method Methods 0.000 title claims abstract description 19
- 239000005556 hormone Substances 0.000 title claims abstract description 10
- 229940088597 hormone Drugs 0.000 title claims abstract description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 13
- 230000001954 sterilising effect Effects 0.000 claims abstract description 10
- 239000001963 growth medium Substances 0.000 claims abstract description 8
- 238000005406 washing Methods 0.000 claims abstract description 4
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 18
- NLKNQRATVPKPDG-UHFFFAOYSA-M potassium iodide Chemical compound [K+].[I-] NLKNQRATVPKPDG-UHFFFAOYSA-M 0.000 claims description 18
- 241000196324 Embryophyta Species 0.000 claims description 17
- NWBJYWHLCVSVIJ-UHFFFAOYSA-N N-benzyladenine Chemical compound N=1C=NC=2NC=NC=2C=1NCC1=CC=CC=C1 NWBJYWHLCVSVIJ-UHFFFAOYSA-N 0.000 claims description 17
- 230000006698 induction Effects 0.000 claims description 13
- 238000005286 illumination Methods 0.000 claims description 12
- 239000006870 ms-medium Substances 0.000 claims description 10
- 235000015097 nutrients Nutrition 0.000 claims description 10
- 238000004659 sterilization and disinfection Methods 0.000 claims description 10
- 230000004069 differentiation Effects 0.000 claims description 9
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 9
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 9
- PAWQVTBBRAZDMG-UHFFFAOYSA-N 2-(3-bromo-2-fluorophenyl)acetic acid Chemical compound OC(=O)CC1=CC=CC(Br)=C1F PAWQVTBBRAZDMG-UHFFFAOYSA-N 0.000 claims description 6
- 239000003109 Disodium ethylene diamine tetraacetate Substances 0.000 claims description 6
- ZGTMUACCHSMWAC-UHFFFAOYSA-L EDTA disodium salt (anhydrous) Chemical compound [Na+].[Na+].OC(=O)CN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O ZGTMUACCHSMWAC-UHFFFAOYSA-L 0.000 claims description 6
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims description 6
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 claims description 6
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 claims description 6
- 239000004327 boric acid Substances 0.000 claims description 6
- GVPFVAHMJGGAJG-UHFFFAOYSA-L cobalt dichloride Chemical compound [Cl-].[Cl-].[Co+2] GVPFVAHMJGGAJG-UHFFFAOYSA-L 0.000 claims description 6
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 claims description 6
- 235000019301 disodium ethylene diamine tetraacetate Nutrition 0.000 claims description 6
- SURQXAFEQWPFPV-UHFFFAOYSA-L iron(2+) sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Fe+2].[O-]S([O-])(=O)=O SURQXAFEQWPFPV-UHFFFAOYSA-L 0.000 claims description 6
- 239000007788 liquid Substances 0.000 claims description 6
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 6
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 6
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 6
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 claims description 6
- LXNHXLLTXMVWPM-UHFFFAOYSA-N pyridoxine Chemical compound CC1=NC=C(CO)C(CO)=C1O LXNHXLLTXMVWPM-UHFFFAOYSA-N 0.000 claims description 6
- 239000011684 sodium molybdate Substances 0.000 claims description 6
- 235000015393 sodium molybdate Nutrition 0.000 claims description 6
- TVXXNOYZHKPKGW-UHFFFAOYSA-N sodium molybdate (anhydrous) Chemical compound [Na+].[Na+].[O-][Mo]([O-])(=O)=O TVXXNOYZHKPKGW-UHFFFAOYSA-N 0.000 claims description 6
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 claims description 6
- 239000011686 zinc sulphate Substances 0.000 claims description 6
- 235000009529 zinc sulphate Nutrition 0.000 claims description 6
- 239000008239 natural water Substances 0.000 claims description 4
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 3
- 239000004471 Glycine Substances 0.000 claims description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 3
- 239000001110 calcium chloride Substances 0.000 claims description 3
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 3
- ZCCIPPOKBCJFDN-UHFFFAOYSA-N calcium nitrate Inorganic materials [Ca+2].[O-][N+]([O-])=O.[O-][N+]([O-])=O ZCCIPPOKBCJFDN-UHFFFAOYSA-N 0.000 claims description 3
- 239000011790 ferrous sulphate Substances 0.000 claims description 3
- 235000003891 ferrous sulphate Nutrition 0.000 claims description 3
- 239000000835 fiber Substances 0.000 claims description 3
- 229940064880 inositol 100 mg Drugs 0.000 claims description 3
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims description 3
- 239000011702 manganese sulphate Substances 0.000 claims description 3
- 235000007079 manganese sulphate Nutrition 0.000 claims description 3
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 3
- 229960003512 nicotinic acid Drugs 0.000 claims description 3
- 235000001968 nicotinic acid Nutrition 0.000 claims description 3
- 239000011664 nicotinic acid Substances 0.000 claims description 3
- 239000004323 potassium nitrate Substances 0.000 claims description 3
- 235000010333 potassium nitrate Nutrition 0.000 claims description 3
- RADKZDMFGJYCBB-UHFFFAOYSA-N pyridoxal hydrochloride Natural products CC1=NC=C(CO)C(C=O)=C1O RADKZDMFGJYCBB-UHFFFAOYSA-N 0.000 claims description 3
- DPJRMOMPQZCRJU-UHFFFAOYSA-M thiamine hydrochloride Chemical compound Cl.[Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N DPJRMOMPQZCRJU-UHFFFAOYSA-M 0.000 claims description 3
- 239000011573 trace mineral Substances 0.000 claims description 3
- 235000013619 trace mineral Nutrition 0.000 claims description 3
- 235000019158 vitamin B6 Nutrition 0.000 claims description 3
- 239000011726 vitamin B6 Substances 0.000 claims description 3
- 229940011671 vitamin b6 Drugs 0.000 claims description 3
- 238000009395 breeding Methods 0.000 claims description 2
- 230000001488 breeding effect Effects 0.000 claims description 2
- 238000011534 incubation Methods 0.000 claims description 2
- 230000008569 process Effects 0.000 claims description 2
- 238000011084 recovery Methods 0.000 claims description 2
- 238000003911 water pollution Methods 0.000 claims description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 abstract description 2
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 abstract description 2
- 230000001902 propagating effect Effects 0.000 abstract 2
- 229930192334 Auxin Natural products 0.000 abstract 1
- 239000005708 Sodium hypochlorite Substances 0.000 abstract 1
- 239000002363 auxin Substances 0.000 abstract 1
- 238000012258 culturing Methods 0.000 abstract 1
- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 abstract 1
- 239000003226 mitogen Substances 0.000 abstract 1
- 239000005972 6-Benzyladenine Substances 0.000 description 12
- JTEDVYBZBROSJT-UHFFFAOYSA-N indole-3-butyric acid Chemical compound C1=CC=C2C(CCCC(=O)O)=CNC2=C1 JTEDVYBZBROSJT-UHFFFAOYSA-N 0.000 description 7
- VGKONPUVOVVNSU-UHFFFAOYSA-N naphthalen-1-yl acetate Chemical compound C1=CC=C2C(OC(=O)C)=CC=CC2=C1 VGKONPUVOVVNSU-UHFFFAOYSA-N 0.000 description 4
- 230000001568 sexual effect Effects 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 230000007226 seed germination Effects 0.000 description 2
- 241000192700 Cyanobacteria Species 0.000 description 1
- 241000756999 Potamogetonaceae Species 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000001066 destructive effect Effects 0.000 description 1
- 238000012851 eutrophication Methods 0.000 description 1
- 239000013505 freshwater Substances 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- YGGXZTQSGNFKPJ-UHFFFAOYSA-N methyl 2-naphthalen-1-ylacetate Chemical compound C1=CC=C2C(CC(=O)OC)=CC=CC2=C1 YGGXZTQSGNFKPJ-UHFFFAOYSA-N 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 244000038651 primary producers Species 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000007634 remodeling Methods 0.000 description 1
- 239000013535 sea water Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
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- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention provides a hormone-induced rapid propagation method of potamogeton pectinatus engineering seedlings. The hormone-induced rapid propagation method mainly comprises the following steps: (1) collecting potamogeton pectinatus, washing away surface attachments with water, and sterilizing the stems by using 75% alcohol and 5% sodium hypochlorite; (2) preparing a culture medium and a culture solution, and setting culture conditions; (3) cultivating new plants: culturing the potamogeton pectinatus stems in the culture medium or the culture solution added with auxin and mitogen to induce the stems to germinate and root, and then transplanting the new potamogeton pectinatus plants with roots and buds into a seedling box for exercising the seedlings. The hormone-induced rapid propagation method provided by the invention is simple to operate, can be used for propagating a large number of potamogeton pectinatus germchits within a short period of time to solve the problem of low reproduction rate of the potamogeton pectinatus, and can be used for continuously propagating the potamogeton pectinatus germchits in the whole year.
Description
Technical field
The present invention relates to a kind of plant cultivation field, particularly relate to a kind of method of hormone induction Fast-propagation Potamogeton pectinatus L engineering seedling.
Background technology
Current lake pollution is serious, and most of water body presents eutrophication, and blue-green alga bloom breaks out, and water transparency obviously declines, and causes destructive destruction to aquatic vegetation.But submerged plant is as the primary producer of lake ecosystem, play a significant role in the circulation of Lakes in Nutrition salt, algal inhibition etc.Therefore lake aquatic vegetation recovers extremely urgent.
Potamogeton pectinatus L is cosmopolitan, and ecological amplitude is wider, can grow, and all have growth in oligotrophic to super eutrophic water body in fresh water and seawater.Therefore Potamogeton pectinatus L can tolerate higher nutritive salt, simultaneously also stronger to the tolerance of blue-green algae.Potamogeton pectinatus L can adapt to various water environment, therefore the suitable key species repaired as lake.But the seed germination of Potamogeton pectinatus L is low, its group is made to recover to there is certain drawback, therefore we have studied Potamogeton pectinatus L utilizes trophosome to carry out the method for Fast-propagation, can set up fairly large group in a short time, makes it become the important population of lake reparation fast.
Potamogeton pectinatus L is also known as fennelleaf pondweed herb, and have sexual propagation and vegetative propagation two kinds of modes, the seed germination rate of wherein sexual propagation mode is low, and less than 6%, therefore vegetative propagation becomes the main modes of reproduction of Potamogeton pectinatus L.Vegetative propagation is bred mainly through stubble, root-like stock, stem tuber, basistipe on the ground etc.Vitro Plant Fast-propagation utilizes the totipotency of plant cell to the tissue such as stem apex, tender shoots, is sprouted and takes root, thus form new plant by cultured in vitro induction.Have and can breed the whole year, be not subject to the restriction of season and weather.Therefore the stem of Potamogeton pectinatus L can be utilized to carry out Fast-propagation, generate more plant.
Summary of the invention
Goal of the invention: in order to improve the proliferative speed of Potamogeton pectinatus L, we utilize plant explants Fast-propagation mode, using the stem section of Potamogeton pectinatus L as research object, adopt the mode of hormone induction to promote that it is sprouted fast, take root.A kind of method promoting Potamogeton pectinatus L Fast-propagation is provided, is conducive to the drawback making up its sexual propagation ability.
In order to achieve the above object, by the following technical solutions, the present invention includes following steps:
(1) sterilization of plant corpus
Gather well-grown, healthy and strong Potamogeton pectinatus L plant, by clean water 3 ~ 4 times, remove surface attachments, leaf is removed.Then to the sterilization of axis section, first use alcohol-pickled 5 minutes of 75%, then soak 10 minutes with the liquor natrii hypochloritis of 5%, finally use aseptic water washing 4 ~ 5 times, each 2 ~ 3 minutes, finally intercept the stem section that 0.5 ~ 0.8cm is with internode.
(2) Fiber differentiation
Axis section after sterilization is moved into the MS medium that with the addition of the IBA of 6-BA and 0.1mg/L of 0.5 ~ 2mg/L, arrange 20 ~ 25 DEG C, intensity of illumination 2000 ~ 3000Lux, illumination 12h/d, cultivate 10 ~ 15 days, induction is sprouted.
Often liter of MS medium contains: ammonium nitrate 1650mg, potassium nitrate 1900mg, calcium chloride 440mg, magnesium sulfate 370mg, potassium dihydrogen phosphate 170mg, potassium iodide 0.83mg, boric acid 6.2mg, magnesium sulfate 22.3mg, zinc sulphate 8.6mg, sodium molybdate 0.25mg, copper sulphate 0.025mg, cobalt chloride 0.025mg, ferrous sulfate 27.8mg, disodium ethylene diamine tetraacetate 37.3mg, inositol 100mg, nicotinic acid 0.5mg, puridoxine hydrochloride (vitamin B6) 0.5mg, thiamine hydrochloride (vitamin B1) 0.1mg, glycine 2.0mg.
(3) Multiplying culture
20 ~ 25 days are cultivated, to obtain the bud of more Potamogeton pectinatus L in the culture access differentiation that will obtain through step (2) and proliferated culture medium.Differentiation and proliferated culture medium are the IBA adding 6-BA and 0.1mg/L of 1.0mg in often liter of MS medium.
(4) culture of rootage
The Potamogeton pectinatus L bud that step (3) obtains to be transferred root induction in culture of rootage liquid, cultivate and can obtain Potamogeton pectinatus L seedling in 10 ~ 15 days, culture of rootage liquid is the Huo Gelan nutrient solution of the 6-BA of NAA and 0.02mg/L that with the addition of 0.5 ~ 2mg/L.
The formula of Huo Gelan nutrient solution:
Contain in often liter of nutrient solution: four water-calcium nitrate 945mg, ammonium nitrate 80mg, potassium dihydrogen phosphate 136mg, magnesium sulfate 493mg, ferrous sulfate heptahydrate 2.78g, disodium ethylene diamine tetraacetate 3.73g; Trace element: potassium iodide 0.00415mg, boric acid 0.031mg, manganese sulphate 0.1115mg, zinc sulphate 0.043mg, sodium molybdate 0.00125mg, copper sulphate 0.000125mg, cobalt chloride 0.000125mg.
(5) hardening
By the Potamogeton pectinatus L seedling that step (4) obtains, be transferred in seedling growing box and cultivate under natural conditions, plant to be planted grows to 20 ~ 30cm can be transplanted to natural water.
Embodiment
By the method for hormone induction Potamogeton pectinatus L Fast-propagation, its concrete implementing measure is as follows:
Well-grown, healthy and strong Potamogeton pectinatus L plant is gathered from natural water.
Then with clear water soak, cleaning 3 ~ 4 times, remove surface attachments, leaf remove after, with 75% alcohol and 5% clorox to the disinfection of stem section, stem section is cut into 0.5 ~ 0.8cm band internode segment.
The MS medium above-mentioned stem section being put into the indolebutyric acid (IBA) of 6-benzyladenine (6-BA) and the 0.1mg/L that with the addition of 0.5 ~ 2mg/L is cultivated, control temperature is at 20 ~ 25 DEG C, illumination 2000 ~ 3000Lux, periodicity of illumination is 12h/d, cultivate 10 ~ 15 days under condition, grow new bud;
Treat that newborn bud grows to 2cm, be transferred in subculture multiplication medium, squamous subculture produces more bud;
The new strain of sprouting is put into the Huo Gelan nutrient solution that with the addition of 0.5 ~ 2mg/L methyl α-naphthyl acetate (NAA) and 0.02mg/L 6-benzyladenine (6-BA) to cultivate about 15 days, grow new root;
The new plant sprouted taking root moves into and is covered with hardening in the seedling growing box of sandstone, transplants to target water body when Potamogeton pectinatus L plant exceeds 30cm.
A method for hormone induction Fast-propagation Potamogeton pectinatus L engineering seedling, comprises the steps:
(1) sterilization of plant corpus
Gather well-grown, healthy and strong Potamogeton pectinatus L plant, by clean water 3 ~ 4 times, remove surface attachments, leaf is removed; Then to the sterilization of axis section, first use alcohol-pickled 5 minutes of 75%, then soak 10 minutes with the liquor natrii hypochloritis of 5%, finally use aseptic water washing 4 ~ 5 times, each 2 ~ 3 minutes, finally intercept the stem section that 0.5 ~ 0.8cm is with internode;
(2) Fiber differentiation
Axis section after sterilization is moved into the MS medium that with the addition of the IBA of 6-BA and 0.1mg/L of 0.5 ~ 2mg/L, arrange 20 ~ 25 DEG C, intensity of illumination 2000 ~ 3000Lux, illumination 12h/d, cultivate 10 ~ 15 days, induction is sprouted;
Often liter of MS medium contains: ammonium nitrate 1650mg, potassium nitrate 1900mg, calcium chloride 440mg, magnesium sulfate 370mg, potassium dihydrogen phosphate 170mg, potassium iodide 0.83mg, boric acid 6.2mg, magnesium sulfate 22.3mg, zinc sulphate 8.6mg, sodium molybdate 0.25mg, copper sulphate 0.025mg, cobalt chloride 0.025mg, ferrous sulfate 27.8mg, disodium ethylene diamine tetraacetate 37.3mg, inositol 100mg, nicotinic acid 0.5mg, puridoxine hydrochloride (vitamin B6) 0.5mg, thiamine hydrochloride (vitamin B1) 0.1mg, glycine 2.0mg.
(3) Multiplying culture
20 ~ 25 days are cultivated, to obtain the bud of more Potamogeton pectinatus L in the culture access differentiation that will obtain through step (2) and proliferated culture medium.Differentiation and proliferated culture medium are the IBA adding 6-BA and 0.1mg/L of 1.0mg in often liter of MS medium.
(4) culture of rootage
The Potamogeton pectinatus L bud that step (3) obtains to be transferred root induction in culture of rootage liquid, cultivate and can obtain Potamogeton pectinatus L seedling in 10 ~ 15 days, culture of rootage liquid is the Huo Gelan nutrient solution of the 6-BA of NAA and 0.02mg/L that with the addition of 0.5 ~ 2mg/L;
The formula of Huo Gelan nutrient solution:
Contain in often liter of nutrient solution: four water-calcium nitrate 945mg, ammonium nitrate 80mg, potassium dihydrogen phosphate 136mg, magnesium sulfate 493mg, ferrous sulfate heptahydrate 2.78g, disodium ethylene diamine tetraacetate 3.73g; Trace element: potassium iodide 0.00415mg, boric acid 0.031mg, manganese sulphate 0.1115mg, zinc sulphate 0.043mg, sodium molybdate 0.00125mg, copper sulphate 0.000125mg, cobalt chloride 0.000125mg.
(5) hardening
By the Potamogeton pectinatus L seedling that step (4) obtains, be transferred in seedling growing box and cultivate under natural conditions, plant to be planted grows to 20 ~ 30cm can be transplanted to natural water.
The method of described Potamogeton pectinatus L Fast-propagation is the IBA of 6-BA and 0.1mg/L adding 0.5 ~ 2mg/L in incubation of sprouting.
The method of described Potamogeton pectinatus L Fast-propagation, adds the 6-BA of NAA and 0.02mg/L of 0.5 ~ 2mg/L in process of rooting culture.
Described Potamogeton pectinatus L method for quickly breeding, the cultivation of explant needs illumination 2000 ~ 3000Lux, and periodicity of illumination is 12h/d.
The seedling cultivated is utilized to be applied to aquatic vegetation plantation and water pollution improvement recovery project.
As can be seen here, implement the present invention and can cultivate a large amount of Potamogeton pectinatus L seedling in a short time.
The above is only preferred embodiment of the present invention, and be not restriction the present invention being made to other form, any those skilled in the art may utilize the technology contents of above-mentioned announcement to be changed or be modified as the Equivalent embodiments of equivalent variations.But everyly do not depart from technical solution of the present invention content, any simple modification, equivalent variations and the remodeling done above embodiment according to technical spirit of the present invention, still belong to the protection domain of technical solution of the present invention.
Claims (5)
1. a method for hormone induction Fast-propagation Potamogeton pectinatus L engineering seedling, is characterized in that, comprise the steps:
(1) sterilization of plant corpus
Gather well-grown, healthy and strong Potamogeton pectinatus L plant, by clean water 3 ~ 4 times, remove surface attachments, leaf is removed; Then to the sterilization of axis section, first use alcohol-pickled 5 minutes of 75%, then soak 10 minutes with the liquor natrii hypochloritis of 5%, finally use aseptic water washing 4 ~ 5 times, each 2 ~ 3 minutes, finally intercept the stem section that 0.5 ~ 0.8cm is with internode;
(2) Fiber differentiation
Axis section after sterilization is moved into the MS medium that with the addition of the IBA of 6-BA and 0.1mg/L of 0.5 ~ 2mg/L, arrange 20 ~ 25 DEG C, intensity of illumination 2000 ~ 3000Lux, illumination 12h/d, cultivate 10 ~ 15 days, induction is sprouted;
Often liter of MS medium contains: ammonium nitrate 1650mg, potassium nitrate 1900mg, calcium chloride 440mg, magnesium sulfate 370mg, potassium dihydrogen phosphate 170mg, potassium iodide 0.83mg, boric acid 6.2mg, magnesium sulfate 22.3mg, zinc sulphate 8.6mg, sodium molybdate 0.25mg, copper sulphate 0.025mg, cobalt chloride 0.025mg, ferrous sulfate 27.8mg, disodium ethylene diamine tetraacetate 37.3mg, inositol 100mg, nicotinic acid 0.5mg, puridoxine hydrochloride (vitamin B6) 0.5mg, thiamine hydrochloride (vitamin B1) 0.1mg, glycine 2.0mg.
(3) Multiplying culture
20 ~ 25 days are cultivated, to obtain the bud of more Potamogeton pectinatus L in the culture access differentiation that will obtain through step (2) and proliferated culture medium.Differentiation and proliferated culture medium are the IBA adding 6-BA and 0.1mg/L of 1.0mg in often liter of MS medium.
(4) culture of rootage
The Potamogeton pectinatus L bud that step (3) obtains to be transferred root induction in culture of rootage liquid, cultivate and can obtain Potamogeton pectinatus L seedling in 10 ~ 15 days, culture of rootage liquid is the Huo Gelan nutrient solution of the 6-BA of NAA and 0.02mg/L that with the addition of 0.5 ~ 2mg/L;
The formula of Huo Gelan nutrient solution:
Contain in often liter of nutrient solution: four water-calcium nitrate 945mg, ammonium nitrate 80mg, potassium dihydrogen phosphate 136mg, magnesium sulfate 493mg, ferrous sulfate heptahydrate 2.78g, disodium ethylene diamine tetraacetate 3.73g; Trace element: potassium iodide 0.00415mg, boric acid 0.031mg, manganese sulphate 0.1115mg, zinc sulphate 0.043mg, sodium molybdate 0.00125mg, copper sulphate 0.000125mg, cobalt chloride 0.000125mg.
(5) hardening
By the Potamogeton pectinatus L seedling that step (4) obtains, be transferred in seedling growing box and cultivate under natural conditions, plant to be planted grows to 20 ~ 30cm can be transplanted to natural water.
2. the method for Potamogeton pectinatus L Fast-propagation as claimed in claim 1, adds the IBA of 6-BA and 0.1mg/L of 0.5 ~ 2mg/L in the incubation that it is characterized in that sprouting.
3. the method for Potamogeton pectinatus L Fast-propagation as claimed in claim 1, is characterized in that the 6-BA of NAA and 0.02mg/L adding 0.5 ~ 2mg/L in process of rooting culture.
4. Potamogeton pectinatus L method for quickly breeding as claimed in claim 1, it is characterized in that the cultivation of explant needs illumination 2000 ~ 3000Lux, periodicity of illumination is 12h/d.
5. the seedling utilizing claim 1 to cultivate is applied to aquatic vegetation plantation and recovery project is administered in water pollution.
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Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
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CN106954549A (en) * | 2017-02-27 | 2017-07-18 | 水生藻安生物科技(武汉)有限公司 | A kind of comb tooth bog pondweed tissue culture and rapid proliferation method |
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CN106954549A (en) * | 2017-02-27 | 2017-07-18 | 水生藻安生物科技(武汉)有限公司 | A kind of comb tooth bog pondweed tissue culture and rapid proliferation method |
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CN107333658A (en) * | 2017-09-13 | 2017-11-10 | 中冶华天工程技术有限公司 | A kind of quick-breeding method of the close thorn eel grass engineering seedling of hormone induction |
CN109601156A (en) * | 2019-01-16 | 2019-04-12 | 中国水产科学研究院珠江水产研究所 | A kind of comb tooth bog pondweed cuttage propagation method |
CN110999773A (en) * | 2019-12-27 | 2020-04-14 | 福建省农业科学院农业生态研究所 | Method for inducing azolla nodularia by using nutrient element P |
CN111631136A (en) * | 2020-07-14 | 2020-09-08 | 安顺职业技术学院 | Tissue culture and rapid propagation method of dormant buds of eyedrops |
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