CN1701661A - Curly pondweed rigid bud tissue culturing method - Google Patents

Curly pondweed rigid bud tissue culturing method Download PDF

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Publication number
CN1701661A
CN1701661A CN200510065450.XA CN200510065450A CN1701661A CN 1701661 A CN1701661 A CN 1701661A CN 200510065450 A CN200510065450 A CN 200510065450A CN 1701661 A CN1701661 A CN 1701661A
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China
Prior art keywords
bud
water
medium
water caltrop
curly pondweed
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CN200510065450.XA
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Chinese (zh)
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CN1303877C (en
Inventor
安树青
郑丽屏
蒋金辉
姚鑫
黄成�
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Nanjing University
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Nanjing University
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Publication of CN1303877C publication Critical patent/CN1303877C/en
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Abstract

The invention relates to the technology field of plant tissue culture, wherein potamogeton crispus turions tissue culture and rapid multiplication technology technique is employed for the mass production of high grade potamogeton crispus tissue culture sprouts, which has overcome the problem of different seed germination under natural conditions, thus meeting the need of recovering aquatic vegetation, developing herbivorous fishes, purifying water and introducing seeds into lakes and ponds.

Description

Curly pondweed rigid bud tissue culturing method
One, technical field
The invention belongs to field of plant tissue culture technique.
Two, background technology
China's aquatic ecosystem seriously polluted, the decline of quality of water environment has caused heavy damage to living aquatic resources, recover the balance of aquatic ecosystem, purifies water, and promotes fisheries development, introduces a fine variety the water body purification plant, and is significant.
Water caltrop (Potamogeton crispus L.) is perennial heavy this plant of pasture and water of Potamogetonaceae (Potamogetonaceae), and its nutritive value and economic worth are bigger.Water caltrop has stronger adaptive capacity to the waters eutrophication, also can densely grow in than the water body of severe contamination by sanitary sewage, has the effect of purifying water body, is the main water plants that purifies water during winter to early summer.But extremely difficulty is sprouted under field conditions (factors), the clint growth is subject to seasonal restrictions and germination rate is low owing to seed, is difficult to large-scale breeding and promotes.
Still the relevant report of not having the research of water caltrop tissue culture technique at present both at home and abroad.
Three, summary of the invention
The objective of the invention is: the technology of inventing a kind of large-scale production water caltrop high-quality tissue cultivating seedling, extremely difficulty is sprouted under field conditions (factors), the clint growth is subject to seasonal restrictions and germination rate is low to overcome this plant seed, the drawback that the retardance large-scale breeding is promoted, adaptation China recovers the breed of aquatic vegetation, development herbivorous fishes or crab class and purifies water, what introduced a fine variety in lake and pond presses for, and promotes China's sustainable development of fishery.
Technical scheme of the present invention is:
Utilize curly pondweed rigid bud to carry out adventitious bud induction culture,, can make its adventitious bud inducing frequency be increased to 100% by hormone regulating and controlling.
Can the foundation of sterile system be to determine it carry out the key of induction smoothly.Improve explant sterilization treatment conditions and medium state by adjustment, obtaining success aspect brownization of checking explant and the pollution, impel normally carrying out of its shoot proliferation, thereby obtain the indefinite bud of breeding fast.
The present invention has set up the fast numerous system of water caltrop high-quality, changes differentiation over to from clint, and plant regeneration frequency is 100%.
On the basis of above research, the propagation method that this invention has been set up the water caltrop indefinite bud is as follows:
(1) at the evoking adventive bud that adds under BA and the KT hormone condition by water caltrop explant clint;
(2) adding under the BA hormone condition, carrying out the successive transfer culture of indefinite bud;
(3) adding under the NAA hormone condition, turn out the complete regeneration plant of taking root
This propagation method stable performance, test good reproducibility, test-tube plantlet well differentiated.
The culture medium prescription of each cultivation stage and hormone combination such as following table:
Curly pondweed rigid bud is cultivated the culture medium prescription in each stage
The group training stage The medium constituent
Adventitious bud inducing ??BA0.5mg/L,KT1.0mg/L
The callus subculture ??BA0.3mg/L
Plant takes root ??NAA0.1mg/L
Effect of the present invention is: be that explant adventitious bud inducing frequency reaches 100% with the clint; The rooting rate of test-tube plantlet reaches more than 90%.
Inducing culture research by clint, the test-tube plantlet of large-scale production water caltrop, and, be beneficial to promotion development with the agronomy quality of technological means such as tissue culture improvement water caltrop.If any enough investments, use this technical scheme, can produce in year more than 1,000,000 strains of water caltrop test-tube plantlet.The water caltrop economic worth is bigger, at first because of its herbaceous stem tender crisp be the good bait of herbivorous fishes, clint contains that abundant vitamin b3 helps the raising of yolk quality and as the feeds of birds such as duck, the water caltrop part of growing thickly is that the good habitat of crab class and shelling phase and crab grow sheltered situation and the first-class farmland green manure of phase; Next is the tool certain nutrient value: thick protein accounts for 26.72% in the content of material, and crude fat accounts for 5.48%, and raw fiber accounts for 15.78%, and nitrogen free extract accounts for 35.67%.The more important thing is that water caltrop has stronger adaptive capacity to the waters eutrophication, also can densely grow in than the water body of severe contamination by sanitary sewage, have the effect of purifying water body, is the main water plants that purifies water during winter to early summer.Have good feed concurrently, recover the characteristics of aquatic vegetation and purifying water body, the demand side that makes it to develop in lake and pond continuable fishery system will be accomplished something.
Four, embodiment
1. set up sterile system
After curly pondweed rigid bud cleaned with running water, in 70% ethanol, soak 30s, use 0.1%HgCl 2The aqueous solution contains a small amount of Tween 80, and sterilization 9min is again with sterile distilled water washing 5 times.Clint after the sterilization is inoculated on the MS medium that does not add exogenous hormone of additional 3% sucrose, being 25 ± 2 ℃ in temperature is 2000lx with light intensity, sets up sterile system under the condition of illumination every day 12h.
2. curly pondweed rigid bud tissue culturing method
Utilize curly pondweed rigid bud to carry out adventitious bud induction culture,, can make its adventitious bud inducing frequency be increased to 100% by hormone regulating and controlling.
Can the foundation of sterile system be to determine it carry out the key of induction smoothly.Improve explant sterilization treatment conditions and medium state by adjustment, obtaining success aspect brownization of checking explant and the pollution, impel normally carrying out of its shoot proliferation, thereby obtain the indefinite bud of breeding fast.
Specific implementation method is as follows:
(1) adventitious bud inducing
Aseptic clint explant is cut into the segment about 3cm, is seeded in to add BA0.5mg/L, and on the MS medium of KT1.0mg/L, be that 25 ± 2 ℃, light intensity are the adventitious bud inducing that 2000lx, every sight are carried out water caltrop explant clint under according to the 12h condition in temperature.
(2) successive transfer culture of indefinite bud
The clump shape indefinite bud that induces is separated into Xiao Cong, on the MS medium that adds BA 0.3mg/L, carries out the successive transfer culture of indefinite bud.
(3) culture of rootage
Clump shape indefinite bud separates and is cut into single plant about 3cm, adding under the NAA 0.1mg/L condition, turns out the complete regeneration plant of taking root, 25 ± 2 ℃ of condition of culture temperature and light intensity 2000lx, illumination every day 12h.
This propagation method stable performance, test good reproducibility, test-tube plantlet well differentiated.With the clint is that explant adventitious bud inducing frequency reaches 100%; The rooting rate of test-tube plantlet reaches more than 90%.

Claims (3)

1, a kind of curly pondweed rigid bud tissue culturing method is characterized in that carrying out adventitious bud induction culture with curly pondweed rigid bud, turns out complete water caltrop test-tube plantlet, and its propagation method is:
(1) curly pondweed rigid bud is cleaned with running water after, in 70% ethanol, soak 30s, use 0.1%HgCl 2The aqueous solution contains a small amount of Tween 80, and sterilization 9min is again with sterile distilled water washing 5 times.Clint after the sterilization is inoculated on the MS medium that does not add exogenous hormone of additional 3% sucrose, being 25 ± 2 ℃ in temperature is 2000lx with light intensity, sets up sterile system under the condition of illumination every day 12h;
(2) at the MS medium that adds BA 0.5mg/L and KT1.0mg/L, by water caltrop explant clint evoking adventive bud;
(3) add the MS medium of BA0.3mg/L, carry out the successive transfer culture of indefinite bud;
(4) at the MS medium that adds NAA0.1mg/L, turn out the complete regeneration plant of taking root, produce the high-quality test-tube plantlet.
2, the water caltrop that produces according to claim 1 is in the application of preparation fish and bird feed.
3, the application of water caltrop in handling effluent sewage, purifying water body of producing according to claim 1.
CN200510065450.XA 2003-12-08 2003-12-08 Curly pondweed rigid bud tissue culturing method Expired - Fee Related CN1303877C (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN200510065450.XA CN1303877C (en) 2003-12-08 2003-12-08 Curly pondweed rigid bud tissue culturing method

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Application Number Priority Date Filing Date Title
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CNB031010652A Division CN1257642C (en) 2002-02-06 2003-01-09 CMOS image sensor
CNB2003101065594A Division CN1237868C (en) 2003-12-08 2003-12-08 Method for cultivating curly pondweed embryo and curly pondweed karren tissue

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CN1701661A true CN1701661A (en) 2005-11-30
CN1303877C CN1303877C (en) 2007-03-14

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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
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
CN103053422A (en) * 2013-01-09 2013-04-24 成都华都农业发展有限责任公司 Method for reproducing potamogeton perfoliatus
CN103734017A (en) * 2014-01-22 2014-04-23 云南大学 Tissue culturing method for water caltrop
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
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

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
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
CN103053422A (en) * 2013-01-09 2013-04-24 成都华都农业发展有限责任公司 Method for reproducing potamogeton perfoliatus
CN103053422B (en) * 2013-01-09 2014-03-26 成都华都农业发展有限责任公司 Method for reproducing potamogeton perfoliatus
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
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
CN106954549B (en) * 2017-02-27 2019-05-28 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
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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