CN1943324A - Method for fast breeding water caltrop seed and seedling - Google Patents
Method for fast breeding water caltrop seed and seedling Download PDFInfo
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- CN1943324A CN1943324A CNA2006101391351A CN200610139135A CN1943324A CN 1943324 A CN1943324 A CN 1943324A CN A2006101391351 A CNA2006101391351 A CN A2006101391351A CN 200610139135 A CN200610139135 A CN 200610139135A CN 1943324 A CN1943324 A CN 1943324A
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Abstract
The method for rapid propagation of water caltrop seedlings includes the following steps: firstly, collecting water caltrop plant from natural water body or greenhouse, making pretreatment and cleaning its surface, removing plant leaf and root, using the stem portion closed to top, sterilizing completely, cutting stem segment, placing the stem segment on the solid culture medium containing hormone to induce budding, transferring the bud with a certain length to propagation culture medium to make amplification culture, then making rooting culture and seedling naturalization, then planting the seedling in natural water body.
Description
Technical field
The present invention relates to the quick propagating technology of water caltrop seed and seedling, also relate to the method for water caltrop stipes tissue culture.Be applicable to that aquatic vegetation recovers a large amount of seedling demands of engineering, also can be aquatic garden landscape, ornamental waterweed is cultivated that seedling is provided, and provide aseptic seedling for water caltrop research.
Background technology
In recent years, along with the implementation of national sustainable development strategy, and the raising of social development level and people's quality of life, the improvement of water pollution control and quality of water environment has caused the strong interest of the whole society.Country has all worked out relevant plan with local, or has carried out relevant scientific research and engineering project.As Tenth Five-Year Plan Period, the Ministry of Science and Technology as one of 12 major scientific and technological projects of country, has carried out Water Ecological Recovery engineering on Taihu Lake, Wuxi, Kunming Dianchi lake, Shenzhen, Shanghai, Zhenjiang and other places with " water pollution control technology and improvement engineering "; State Environmental Protection Administration has worked out " three rivers " " three lakes " water prevention and cure of pollution the Tenth Five-Year Plan (2001-2005) together with department such as national development and the reform committee and government of relevant provinces, autonomous regions and municipalities, and State Council's reply is implemented." plan " determined 1590 projects altogether, 1,234 hundred million yuan of gross investments.
Water pollute and the measure of the improvement of quality of water environment in, cutting under the dirty prerequisite, rebuild water ecosystem, the self-purification capacity of the aquatic ecosystem of getting well is considered to solve steadily in the long term the essential measure of water pollution.As the key members of aquatic ecosystem, submerged plant not only plays an important role aspect perching water body food web, the supply of water body dissolved oxygen, nutrition circulation, aquatic animal, and has proved that submerged vegetation is a key factor of keeping the clean waters state.Yet the eutrophication of water body and the excessive herbivorous fishes of culturing have caused the extensive degeneration of submerged vegetation, so the recovery of submerged vegetation is the committed step during water pollution is administered.At Dian Chi water pollution control environment-friendly engineering, in the engineering projects such as country's major scientific and technological project " area, Hanyang, Wuhan City quality of water environment improves technology and comprehensive demonstration ", " Taihu Lake water pollution control and water body recovery technique and engineering demonstration ", the recovery of submerged vegetation all is a key content wherein.
The recovery engineering of aquatic vegetation must be brought the wilderness demand to the water plants seedling.Usually every square metre of water surface needs 5-10 strain (root and stem of certain plants) water plants seedling, and every square kilometre of water surface then needs the up to ten million strains of seedling (root and stem of certain plants), even every strain seedling is pressed 0.2 yuan of calculating, the expense of every square kilometre of required seedling of the water surface also reaches 2,000,000 yuan.The seedling that uses in the recovery engineering of aquatic vegetation at present is to fish for from natural water fully, the way that this method comes down to rob Peter to pay Paul, not only the water plants resource to source of seedling ground damages, in fact also often run into the shortage of seedling resource, promptly can't obtain enough seedling amounts of certain plants.In addition, the acquisition of water plants seedling also usually is subjected to the influence in season.Therefore, in order to satisfy the active demand of water plants seedling, be badly in need of the new source mode of the aquatic plant seedling of development.The approach of breeding water plants fast by the mode of Plant Tissue Breeding is a kind of production method of feasible fully water plants seedling.Plant Tissue Breeding is by the part of sterile working separating plant body, is inoculated into medium, develops into the process of whole plant under the manual control condition.This class mode has been widely used in the fast seedling growing of crop, medicinal plant, economy forest and famous-brand and high-quality flowers.This method can go out a large amount of seedlings from number strain plant production at short notice, and not only floor space is little, and is not subjected to seasonal effect, and the seedling of producing has been removed plant virus, and proterties is more good.Aspect the seedling of economy plant, the home and abroad all has a large amount of companies to be engaged in commercial production, and it is numerous soon to carry out orchid as the many areas in the U.S., Europe and Southeast Asia with tissue culture method, has become famous " orchid industry ".Aspect ecological recovery, existing three the commercial laboratories of the U.S. utilize this mode to produce the recovery engineering that hygrophyte is used for wetlands ecosystems.
Water caltrop is the common submerged plant of China's shallow lake, and durability against pollution is preferably arranged, the pionner of Chang Zuowei aquatic vegetation reconstruction engineering first-selection.Because the epidermis of submerged plant is strong to the permeability of bactericide, be that the tissue culture of explant is more more difficult than terrestrial plant therefore with root, stem, leaf.External at present report submerged plant watermifoil and the training of Potamogeton pectinatus L group are set up.Yet there are no report with the water caltrop stem as the tissue-culturing rapid propagation of explant.
Summary of the invention
The object of the present invention is to provide a kind of method of fast breeding water caltrop seed and seedling, effectively solve a large amount of supply and demand problems of water caltrop seed and seedling, material is easy to obtain, process fast, is not subjected to seasonal effect, with low cost.
In order to achieve the above object, the present invention has adopted following technical scheme:
1, gather in the natural water or the water caltrop seedling of the robust growth of cultivating in the greenhouse;
2, with running water flushing 1-2 hour, defoliation and root were got the sturdy stem on top, use distilled water flushing 2-3 time, clean the water caltrop stem;
3, inducing of sterilization and bud: adopt bactericidal agent PPM (Plant preserve Mixture), ethanol, clorox and hydrogen peroxide etc. to the degerming of stem explant.Its degerming mode is for 50% ethanol sterilization 1 minute, then with sterile distilled water flushing three times, again with 1/100 milliliters sterilizations of 1.05%NaClO+ Tween-20 10-20 minute, at last with sterile distilled water flushing three times; Or 5%H
2O
2Sterilized 15 minutes, and with sterile distilled water flushing three times, used 4-10%PPM15-20 minute more then, at last with sterile distilled water flushing three times.The stem of bacterium of going out soaked 4-6 minute with sterile distilled water, the stem that cuts 5-8 millimeter band internode with scalpel is 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+15-30g/L 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-EDTA is 37.3mg/L, FeSO
4.4H
2O is 27.8mg/L;
Vitamin: 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, 2ip, IBA, NAA
Induce axillalry bud to sprout and the hormone combination of differentiation adventitious buds in the medium:
Add 6-BA 1.0-5.0mgL
-1+ IBA 0.5mgL
-1Or
6-BA 1.0-5.0mgL
-1+ NAA 0.5mgL
-1Or
2ip 1.0-5.0mgL
-1+ IBA 0.5mgL
-1Or
2ip?1.0-5.0mg·L
-1+NAA?0.5mg·L
-1；
(3) medium proportioning mode is:
MS+15-30g/L sucrose+0.8% agar+hormone or
MS+0.8% agar+hormone or
MS/5+15-30g/L sucrose+0.8% agar+hormone or
MS/5+0.8% agar+hormone or
MS/10+15-30g/L sucrose+0.8% agar+hormone or
MS/10+0.8% agar+hormone;
Medium pH is 5.8-6.0.
4, the propagation of bud: induced sprouting in 6 or 7 or 8 days, treat that bud is long to 1-2 centimetre, be transferred in the medium of inducing propagation and breed, the medium of shoot proliferation is: MS or 1/5MS or 1/10MS+15-30g/L sucrose+0.8% agar+hormone, hormone combination is: 6-BA 1.0-5.0mgL
-1Or 2ip 1.0-5.0mgL
-1
5, inducing of root: when waiting to grow bud, be transferred in the medium of root induction, induce it to take root, the medium of root induction is: MS or 1/5MS or 1/10MS+15-30g/L sucrose+0.8% agar+hormone, hormone combination is: IBA or NAA 1.0-3.0mgL
-1+ 6-BA or 2ip 0.5mgL
-1
When 6, treating that seedling sends out roots, open and seal film, cultivated 2-3 days, be transferred under the natural conditions and cultivate.
The present invention compared with prior art, have the following advantages and effect: 1, can realize that water caltrop breeds on a large scale, avoid in the ecological recovery engineering of water plants when a large amount of seedling of needs by fishing for from natural water, not only the water plants resource to source of seedling ground damages, and also can often run into the shortage of seedling resource simultaneously; 2, be not subjected to the influence in season, from natural water, to fish for plant and be subjected to the influence in season big, the kind shoot survival percent that different time is gathered is also different, can control vegetative period of seedling by industrial production, is convenient to plant in best period; 3, material source is easy, and is with low cost
Description of drawings
Fig. 1 is the healthy and strong seedling of water caltrop, is collected in greenhouse or the outdoor natural water, as the source of explant
The sprouting that Fig. 2 induces in inducing the group training medium of sprouting for water caltrop stipes explant,
Fig. 3 sprouts the bud that bunch for water caltrop sprouting propagation in group training proliferated culture medium
Fig. 4 induces in the group training medium of root induction for the water caltrop bud takes root
The water caltrop complete seedling of Fig. 5 for obtaining through tissue culture
Embodiment
A kind of method of fast breeding water caltrop seed and seedling, specifically implement as follows:
Embodiment 1:
(A) in the winter time or gather the water caltrop seedling of robust growth spring from natural water or in the greenhouse;
(B) in the laboratory with running water flushing 1-2 hour, remove the pollutant on water caltrop surface, treat that the water caltrop seedling is rinsed well after, remove Ye Hegen, cutting is by the sturdy stem on top, with distilled water flushing 2-3 time, thoroughly cleans the water caltrop stem;
(C) inducing of sterilization and bud: the instrument that needs to use in experimentation needs through 121 ℃ of 0.1MPa high pressure steam sterilizations as flat board, tweezers and medium etc., experimental tool will be positioned in Biohazard Safety Equipment or the sterile working platform and sterilize 30 minutes when experimentizing, adopt the bactericidal agent degerming then under aseptic condition, bactericidal agent is respectively PPM-Plant preserve Mixture, ethanol, clorox and hydrogen peroxide.Its degerming mode is for 50% ethanol sterilization 1 minute, then with sterile distilled water flushing three times, again with 1/100 milliliters sterilizations of 1.05%NaClO+ Tween-20 10-20 minute, at last with sterile distilled water flushing three times; Or 5%H
2O
2Sterilized 15 minutes, and with sterile distilled water flushing three times, used 4-10%PPM15-20 minute more then, at last with sterile distilled water flushing three times.The stem of bacterium of going out soaked 4-6 minute with sterile distilled water, the stem that cuts 5-8 millimeter band internode with scalpel is 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+15-30g/L sucrose+0.8% agar+hormone, and its concrete ratio is as follows:
Medium component:
A, 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-EDTA is 37.3mg/L, FeSO
4.4H
2O is 27.8mg/L
Vitamin: 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
B, contained hormone kind: 6-BA, 2ip, IBA, NAA
Induce axillalry bud to sprout in the medium and the hormone combination of differentiation adventitious buds is:
1. 6-BA or 2ip1.0mgL
-1+ IBA or NAA0.5mgL
-1
2. 6-BA or 2ip1.5mgL
-1+ IBA or NAA0.5mgL
-1
3. 6-BA or 2ip2.0mgL
-1+ IBA or NAA0.5mgL
-1
4. 6-BA or 2ip2.5mgL
-1+ IBA or NAA0.5mgL
-1
5. 6-BA or 2ip3.0mgL
-1+ IBA or NAA0.5mgL
-1
6. 6-BA or 2ip3.5mgL
-1+ IBA or NAA0.5mgL
-1
7. 6-BA or 2ip4.0mgL
-1+ IBA or NAA0.5mgL
-1
8. 6-BA or 2ip4.5mgL
-1+ IBA or NAA0.5mgL
-1
9. 6-BA or 2ip5.0mgL
-1+ IBA or NAA0.5mgL
-1
C, medium proportioning mode
MS+15-30g/L sucrose+0.8% agar+hormone or
MS+0.8% agar+hormone or
MS/5+15-30g/L sucrose+0.8% agar+hormone or
MS/5+0.8% agar+hormone or
MS/10+15-30g/L sucrose+0.8% agar+hormone or
MS/10+0.8% agar+hormone
Medium pH is 5.8-6.0.
(D) propagation of bud: vaccinated flat board or blake bottle be positioned in illumination box or the culturing room cultivate, cultivation temperature is 15-25 ℃, intensity of illumination 1000-5000Lux, and light application time is illumination 12-24 hour every day.Induce it to sprout, can sprout sprouting in 6 or 7 or 8 days, treat that bud length to 1-2 centimetre, is transferred in the shoot proliferation medium, the shoot proliferation medium is: MS or 1/5MS or 1/10MS+15-30g/L sucrose+0.8% agar+hormone, hormone combination is: 6-BA 2.5mgL
-1
(E) inducing of root: when waiting to grow bud in 9 or 10 or 11 days, be transferred to root induction in the medium of root induction; Root induction medium: MS or 1/5MS or 1/10MS+15-30g/L sucrose+0.8% agar+hormone, hormone combination is: IBA 1.0mgL
-1+ 6-BA 0.5mgL
-1
When (F) treating that seedling grows more, open and seal film, cultivated 2-3 days, be transplanted to outdoor cultivation under field conditions (factors) then.
Embodiment 2:
The propagation hormone combination of A bud:
2ip?2.5mg·L
-1
The B root induce hormone combination:
IBA?1.0mg·L
-1+2ip?0.5mg·L
-1
Other implementation step is identical with embodiment 1
Embodiment 3:
The propagation hormone combination of A bud:
6-BA?5.0mg·L
-1
The B root induce hormone combination:
NAA?1.0mg·L
-1+6-BA?0.5mg·L
-1
Other implementation step is identical with embodiment 1.
Embodiment 4
The propagation hormone combination of A bud:
2ip?4.0mg·L
-1
The B root induce hormone combination:
NAA?2.0mg·L
-1+2ip?0.5mg·L
-1
Other implementation step is identical with embodiment 1.
Claims (3)
1, a kind of method of fast breeding water caltrop seed and seedling, it comprises the following steps:
A, gather in the natural water or the water caltrop seedling of cultivating in the greenhouse;
B, with running water flushing 1-2 hour, defoliation and root are got the stem by the top, with distilled water flushing 2-3 time, clean the water caltrop stem;
Inducing of C, sterilization and bud: bactericidal agent is that PPM, ethanol, clorox and hydrogen peroxide are to the degerming of stem explant, then with sterile distilled water flushing 2-4 time, soaked 4-6 minute, the stem that cuts 5-8 millimeter band internode is as explant, be inoculated into to induce in the medium of sprouting and cultivate, induce the medium of sprouting to be: MS or 1/5MS or 1/10MS+15-30g/L sucrose+0.8% agar+hormone, hormone combination is: 6-BA or 2ip1.0-5.0mgL
-1+ IBA0.5mgL
-1Or NAA0.5mgL
-1
The propagation of D, bud: vaccinated flat blake bottle is positioned in illumination box or the culturing room to cultivate cultivation temperature be 15-25 ℃, intensity of illumination 1000-5000Lux, light application time is 12-24 hour every day, induced in 6-8 days and sprout, treat that bud is long to 1-2 centimetre, be transferred in the shoot proliferation medium, the shoot proliferation medium is: MS or 1/5MS or 1/10MS+15-30g/L sucrose+0.8% agar+hormone, hormone combination is: 6-BA 1.0-5.0mgL
-1Or 2ip1.0-5.0mgL
-1
E, when waiting to grow bud, be transferred in the medium of root induction, induce it to take root, the medium of root induction is: MS or 1/5MS or 1/10MS+15-30g/L sucrose+0.8% agar+hormone, hormone combination is: IBA or IBA 1.0-3.0mgL
-1+ 6-BA or 2ip 0.5mgL
-1
F, when treating that seedling sends out roots, open and seal film, cultivated 2-3 days, be transplanted to then and outdoorly cultivate under field conditions (factors).
2, the method for a kind of fast breeding water caltrop seed and seedling according to claim 1 is characterized in that:
Bud induce or the proportioning mode of the medium of the propagation of bud or root induction is:
MS+15-30g/L sucrose+0.8% agar+hormone or
MS+0.8% agar+hormone or
MS/5+15-30g/L sucrose+0.8% agar+hormone or
MS/5+0.8% agar+hormone or
MS/10+15-30g/L sucrose+0.8% agar+hormone or
MS/10+0.8% agar+hormone.
3, the method for a kind of fast breeding water caltrop seed and seedling according to claim 1 is characterized in that:
Sterilization is with 50% ethanol sterilization 1 minute, then with sterile distilled water flushing three times, again with 1/100 milliliters sterilizations of 1.05%NaCl0+ Tween-20 10-20 minute, at last with sterile distilled water flushing three times; Or use 5%H
2O
2Sterilized 15 minutes, and with sterile distilled water flushing three times, used 4-10%PPM15-20 minute more then, at last with sterile distilled water flushing three times.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2006101391351A CN100462001C (en) | 2005-10-13 | 2006-10-12 | Method for fast breeding water caltrop seed and seedling |
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200510019580.X | 2005-10-13 | ||
| CNA200510019580XA CN1742563A (en) | 2005-10-13 | 2005-10-13 | Method for fast breeding water caltrop seedlings |
| CNB2006101391351A CN100462001C (en) | 2005-10-13 | 2006-10-12 | Method for fast breeding water caltrop seed and seedling |
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| Publication Number | Publication Date |
|---|---|
| CN1943324A true CN1943324A (en) | 2007-04-11 |
| CN100462001C CN100462001C (en) | 2009-02-18 |
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ID=38043140
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2006101391351A Expired - Fee Related CN100462001C (en) | 2005-10-13 | 2006-10-12 | Method for fast breeding water caltrop seed and seedling |
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|---|---|
| CN (1) | CN100462001C (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101120655B (en) * | 2007-09-18 | 2010-11-17 | 华中师范大学 | Method for making potamogeton cripus artificial seed |
| CN102090182A (en) * | 2010-12-27 | 2011-06-15 | 中国科学院测量与地球物理研究所 | Method for sowing Potamogeton crispus turions |
| CN102450149A (en) * | 2010-11-01 | 2012-05-16 | 南京中科水治理工程有限公司 | Cutting propagation method for potamogeton crispus L |
| CN102557270A (en) * | 2012-03-02 | 2012-07-11 | 江苏美尚生态景观股份有限公司 | Effective method for removing phenol from water by using potamogeton crispus |
| CN103734017A (en) * | 2014-01-22 | 2014-04-23 | 云南大学 | Tissue culturing method for water caltrop |
| CN104620992A (en) * | 2015-03-03 | 2015-05-20 | 佛山市顺德区今日景艺生物科技有限公司 | Rhynchostylis lateral bud induction culture medium and culture method |
| 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 |
Family Cites Families (1)
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| CN1237868C (en) * | 2003-12-08 | 2006-01-25 | 南京大学 | Method for cultivating curly pondweed embryo and curly pondweed karren tissue |
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- 2006-10-12 CN CNB2006101391351A patent/CN100462001C/en not_active Expired - Fee Related
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101120655B (en) * | 2007-09-18 | 2010-11-17 | 华中师范大学 | Method for making potamogeton cripus artificial seed |
| CN102450149A (en) * | 2010-11-01 | 2012-05-16 | 南京中科水治理工程有限公司 | Cutting propagation method for potamogeton crispus L |
| CN102090182A (en) * | 2010-12-27 | 2011-06-15 | 中国科学院测量与地球物理研究所 | Method for sowing Potamogeton crispus turions |
| CN102090182B (en) * | 2010-12-27 | 2012-05-23 | 中国科学院测量与地球物理研究所 | Method for sowing Potamogeton crispus turions |
| CN102557270A (en) * | 2012-03-02 | 2012-07-11 | 江苏美尚生态景观股份有限公司 | Effective method for removing phenol from water by using potamogeton crispus |
| CN103734017A (en) * | 2014-01-22 | 2014-04-23 | 云南大学 | Tissue culturing method for water caltrop |
| CN103734017B (en) * | 2014-01-22 | 2016-04-20 | 云南大学 | The method for tissue culture of a kind of water caltrop |
| CN104620992A (en) * | 2015-03-03 | 2015-05-20 | 佛山市顺德区今日景艺生物科技有限公司 | Rhynchostylis lateral bud induction culture medium and culture method |
| 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 |
| CN111631136B (en) * | 2020-07-14 | 2022-11-25 | 安顺职业技术学院 | Tissue culture and rapid propagation method of dormant buds of eyedrops |
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| Publication number | Publication date |
|---|---|
| CN100462001C (en) | 2009-02-18 |
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