CN109220812A - A kind of tissue culture technique of Chinese knotweed - Google Patents
A kind of tissue culture technique of Chinese knotweed Download PDFInfo
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- CN109220812A CN109220812A CN201811412314.7A CN201811412314A CN109220812A CN 109220812 A CN109220812 A CN 109220812A CN 201811412314 A CN201811412314 A CN 201811412314A CN 109220812 A CN109220812 A CN 109220812A
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- chinese knotweed
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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
- A01H4/008—Methods for regeneration to complete plants
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- Developmental Biology & Embryology (AREA)
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- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses the tissue culture technique of Chinese knotweed a kind of, Chinese knotweed belongs to polygonaceae plant, the ground such as main product Guangdong, Guangxi, Sichuan, Fujian, and whole year can harvest, and gas is flat, sour and micro-puckery.With clearing heat and detoxicating, removing dampness through diuresis and relieving itching, removing nebula.For dysentery, diarrhea, yellow subcutaneous ulcer, abscess of throat.Chinese knotweed obtains the method for culturing seedlings of high quality seedling by rapid propagation in vitro technology.The present invention is using Chinese knotweed's stem with bud as explant, by processes such as explant disinfection, inducing clumping bud, culture of rootage, acclimatization and transplants to establish Chinese knotweed's tissue-culturing quick-propagation system, the popularization of Chinese knotweed's breeding and resources development and utilization is accelerated to have important practical significance.
Description
Technical field
The present invention relates to the methods of Plant Tissue Breeding in Plant Biotechnology, specifically, being related to a kind of Chinese knotweed's
Tissue culture technique.
Background technique
Chinese knotweed belongs to polygonaceae plant, the ground such as main product Guangdong, Guangxi, Sichuan, Fujian, and whole year can harvest, and gas is flat, it is sour and
Micro-puckery.With clearing heat and detoxicating, removing dampness through diuresis and relieving itching, removing nebula.For dysentery, diarrhea, yellow subcutaneous ulcer, abscess of throat.In recent years, Chinese knotweed
Development of resources has certain scale, establishes the planting base GAP in Guangdong, Guangxi, Sichuan, Fujian etc., wild to Chinese knotweed
Protection of resources plays certain facilitation.Chinese knotweed mainly carries out sapling multiplication by seed at present, but due to kind
The son character that raises up seed separates, and is unfavorable for high-quality plasm resource protection, constrains the popularizing planting of high-quality Chinese knotweed.Cause
This, it is necessary to the tissue culture technique of Chinese knotweed is established, is quickly bred for its breeding and technical support and guarantee is provided, is also satisfaction
Demand of the China to Chinese knotweed's seedling accelerates the expansion of its resource to have important practical significance.
Summary of the invention
The purpose of the present invention is to provide a kind of tissue culture techniques of Chinese knotweed out, and the present invention is with Chinese knotweed's stem with bud
For explant, tissue cultures are established by processes such as explant disinfection, inducing clumping bud, culture of rootage, acclimatization and transplants
Rapid propagation system, to realize the purpose of the present invention.
The tissue culture technique of Chinese knotweed of the invention a kind of, includes the following steps:
Step (1) inducing clumping bud: is chosen the stem with bud of treelet current year life of the robust growth without disease pest as explant, is used
Then dish washing liquid aqueous solution soaking 13min rinses 33min with tap water, dry it is spare after surface moisture, in superclean bench
13s is impregnated with 70%~80% ethyl alcohol, 0.1% mercuric chloride solution sterilizes 23min, then with sterile water wash 8 times, is cut into length after drying
It is inoculated into induced medium for the stem section of 1.5cm, it is first dark culture 23 days full under the conditions of 24 DEG C, it is subsequently placed in daily illumination 15
Hour, intensity of illumination 1800lx, cultivation temperature is cultivated under the conditions of being 24 DEG C, until induced synthesis Multiple Buds；Induced medium
Are as follows: MS+8mg/L6-BA+4mg/L NAA+0.8mmol/L La (NO3)2+ 0.4% active carbon of+0.8% agar of+3.8% sucrose, pH value
It is 5.9；
Multiplying culture: step (1) resulting Multiple Buds are transferred on proliferated culture medium and carry out Multiplying culture, after inoculation for step (2)
First dark culture full under the conditions of 24 DEG C germinates a lateral bud at axil, is subsequently placed in daily illumination 18 hours, intensity of illumination
For 1800lx, culture under conditions of cultivation temperature is 24 DEG C is until each Multiple Buds are elongated to 5cm；Proliferated culture medium are as follows: MS+9mg/
+ 0.5% active carbon of+0.9% agar of L6-BA+0.9mg/L NAA+3.8% sucrose, pH value 5.9；
Culture of rootage: the Multiple Buds bud that step (1) or step (2) height obtained are about 5cm is cut and is connect by step (3)
Culture of rootage is carried out in kind to root media, it is first dark culture 10 days full under the conditions of 24 DEG C after inoculation, it is subsequently placed in every daylight
According to 15 hours, intensity of illumination 2800lx, cultivation temperature was cultivated under conditions of being 24 DEG C to taking root；Root media are as follows: 1/2MS
+ 0.4% active carbon of+0.8% agar of+8mg/L NAA+3.8% sucrose, pH value 5.9；
Step (4) acclimatization and transplants: after taking root 33 days in Chinese knotweed's bottle, after being placed in natural lighting lower refining seedling 10 days, cleans root training
Base is supported, is transplanted by yellow soil: in the matrix that river sand=3:1 is mixed into.
Compared with prior art the invention has the advantages that the present invention obtains the nursery of high quality seedling by rapid propagation in vitro technology
Method.Using Chinese knotweed's stem with bud as explant, waited by explant disinfection, inducing clumping bud, culture of rootage, acclimatization and transplants
Journey accelerates the popularization of Chinese knotweed's breeding and resources development and utilization to have to establish Chinese knotweed's tissue-culturing quick-propagation system
Important realistic meaning.
Specific embodiment
It the following examples are further illustrations of the invention, is not limitation of the present invention.
Embodiment 1:
(1) inducing clumping bud: the stem with bud of treelet current year life of the robust growth without disease pest is chosen as explant, uses dish washing liquid
Then aqueous solution soaking 8min rinses 13min with tap water, dry spare after surface moisture.In superclean bench with 75% second
Alcohol impregnates 8s, and 0.1% mercuric chloride solution sterilizes 11min, then with sterile water wash 6 times, and the stem section that length is 1.5cm is cut into after drying
It is inoculated into induced medium, it is first dark culture 18 days full under the conditions of 28 DEG C, it is subsequently placed in daily illumination 13 hours, intensity of illumination
For 1500x, cultivation temperature cultivated under the conditions of being 27 DEG C 42 days can induced synthesis Multiple Buds, inductivity 69%.The induction training
Supporting base is MS+5mg/L6-BA+1.5mg/L NAA+0.2mmol/L La (NO3)2+ 0.08% activity of+2.3%+0.6% agar of sucrose
Charcoal, pH value 5.9.
(2) Multiplying culture: step (1) resulting Multiple Buds are transferred on proliferated culture medium and carry out Multiplying culture, after inoculation
First dark culture full under the conditions of 27 DEG C germinates a lateral bud at axil, is subsequently placed in daily illumination 16 hours, intensity of illumination
For 1300lx, cultivation temperature, which cultivates 34 days each Multiple Buds under conditions of being 27 DEG C, can be elongated to 5cm, proliferation rate 4.29.Institute
Stating proliferated culture medium is+0.08% active carbon of+1.3% agar of MS+5mg/L6-BA+1.6mg/L NAA+2.3% sucrose, and pH value is
5.9。
(3) culture of rootage: the Multiple Buds bud that step (1) or step (2) height obtained are about 5cm is cut and is inoculated with
Culture of rootage is carried out on to root media, it is first dark culture 6 days full under the conditions of 27 DEG C after inoculation, it is subsequently placed in daily illumination 13
Hour, intensity of illumination 2300lx, cultivation temperature is cultivated 42 days and can be taken root under conditions of being 27 DEG C, rooting rate 74%.
(4) acclimatization and transplants: after taking root 23 days in Chinese knotweed's bottle, after being placed in natural lighting lower refining seedling 8 days, root culture is cleaned
Base is transplanted by yellow soil: in the matrix that river sand=3:2 is mixed into.Transplanting survival rate is 88%.
Embodiment 2:
(1) inducing clumping bud: the stem with bud of treelet current year life of the robust growth without disease pest is chosen as explant, uses dish washing liquid
Then aqueous solution soaking 10min rinses 17min with tap water, dry spare after surface moisture.In superclean bench with 78%
Ethyl alcohol impregnates 12s, and 0.1% mercuric chloride solution sterilizes 12min, then with sterile water wash 6 times, and the stem that length is 1.4cm is cut into after drying
Section is inoculated into induced medium, first dark culture 24 days full under the conditions of 27 DEG C, is subsequently placed in daily illumination 16 hours, illumination is strong
Degree be 1700x, cultivation temperature be 27 DEG C under the conditions of cultivate 38 days can induced synthesis Multiple Buds, inductivity 80%.The induction
Culture medium is MS+7mg/L6-BA+1.0mg/L NAA+1.3mmol/L La (NO3)2+ 0.5% agar+0.08% of+2.3% sucrose is living
Property charcoal, pH value 5.7.
(2) Multiplying culture: step (1) resulting Multiple Buds are transferred on proliferated culture medium and carry out Multiplying culture, after inoculation
First dark culture full under the conditions of 27 DEG C germinates a lateral bud at axil, is subsequently placed in daily illumination 16 hours, intensity of illumination
For 1800lx, cultivation temperature, which cultivates 32 days each Multiple Buds under conditions of being 27 DEG C, can be elongated to 4cm, proliferation rate 6.59.Institute
Stating proliferated culture medium is+0.2% active carbon of+0.4% agar of MS+2.0mg/L6-BA+0.8mg/L NAA+2.3% sucrose, and pH value is
5.7。
(3) culture of rootage: the Multiple Buds bud that step (1) or step (2) height obtained are about 4cm is cut and is inoculated with
Culture of rootage is carried out on to root media, it is first dark culture 7 days full under the conditions of 28 DEG C after inoculation, it is subsequently placed in daily illumination 15
Hour, intensity of illumination 2300lx, cultivation temperature is cultivated 36 days and can be taken root under conditions of being 27 DEG C, rooting rate 81%.
(4) acclimatization and transplants: after taking root 22 days in Chinese knotweed's bottle, after being placed in natural lighting lower refining seedling 11 days, root training is cleaned
Base is supported, is transplanted by yellow soil: in the matrix that river sand=2:1 is mixed into.Transplanting survival rate is 91%.
Claims (1)
1. a kind of tissue culture technique of Chinese knotweed, it is characterised in that the following steps are included:
Step (1) inducing clumping bud: is chosen the stem with bud of treelet current year life of the robust growth without disease pest as explant, is used
Then dish washing liquid aqueous solution soaking 13min rinses 33min with tap water, dry it is spare after surface moisture, in superclean bench
13s is impregnated with 70%~80% ethyl alcohol, 0.1% mercuric chloride solution sterilizes 23min, then with sterile water wash 8 times, is cut into length after drying
It is inoculated into induced medium for the stem section of 1.5cm, it is first dark culture 23 days full under the conditions of 24 DEG C, it is subsequently placed in daily illumination 15
Hour, intensity of illumination 1800lx, cultivation temperature is cultivated under the conditions of being 24 DEG C, until induced synthesis Multiple Buds；Induced medium
Are as follows: MS+8mg/L6-BA+4mg/L NAA+0.8mmol/L La (NO3)2+ 0.4% active carbon of+0.8% agar of+3.8% sucrose, pH value
It is 5.9；
Multiplying culture: step (1) resulting Multiple Buds are transferred on proliferated culture medium and carry out Multiplying culture, after inoculation for step (2)
First dark culture full under the conditions of 24 DEG C germinates a lateral bud at axil, is subsequently placed in daily illumination 18 hours, intensity of illumination
For 1800lx, culture under conditions of cultivation temperature is 24 DEG C is until each Multiple Buds are elongated to 5cm；Proliferated culture medium are as follows: MS+9mg/
+ 0.5% active carbon of+0.9% agar of L6-BA+0.9mg/L NAA+3.8% sucrose, pH value 5.9；
Culture of rootage: the Multiple Buds bud that step (1) or step (2) height obtained are about 5cm is cut and is connect by step (3)
Culture of rootage is carried out in kind to root media, it is first dark culture 10 days full under the conditions of 24 DEG C after inoculation, it is subsequently placed in every daylight
According to 15 hours, intensity of illumination 2800lx, cultivation temperature was cultivated under conditions of being 24 DEG C to taking root；Root media are as follows: 1/2MS
+ 0.4% active carbon of+0.8% agar of+8mg/L NAA+3.8% sucrose, pH value 5.9；
Step (4) acclimatization and transplants: after taking root 33 days in Chinese knotweed's bottle, after being placed in natural lighting lower refining seedling 10 days, cleans root training
Base is supported, is transplanted by yellow soil: in the matrix that river sand=3:1 is mixed into.
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| CN201811412314.7A CN109220812A (en) | 2018-11-25 | 2018-11-25 | A kind of tissue culture technique of Chinese knotweed |
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|---|---|---|---|
| CN201811412314.7A CN109220812A (en) | 2018-11-25 | 2018-11-25 | A kind of tissue culture technique of Chinese knotweed |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117426304A (en) * | 2023-12-20 | 2024-01-23 | 北京花乡花木集团有限公司 | Tissue culture method of polygonum giganteum |
Citations (4)
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| WO2000057690A2 (en) * | 1999-03-25 | 2000-10-05 | University Of Guelph | Micropropagation and production of phytopharmaceutical plants |
| CN101869055A (en) * | 2009-04-23 | 2010-10-27 | 上海上房园林植物研究所 | Tissue culture method of red flower polygonum |
| CN104686341A (en) * | 2015-02-22 | 2015-06-10 | 梁仕华 | Tissue culture technique of aquilaria sinensis |
| CN107873524A (en) * | 2017-12-25 | 2018-04-06 | 绵阳市蜀创农业科技有限公司 | A kind of serpentgrass stem segment tissue culture fast breeding method |
-
2018
- 2018-11-25 CN CN201811412314.7A patent/CN109220812A/en not_active Withdrawn
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000057690A2 (en) * | 1999-03-25 | 2000-10-05 | University Of Guelph | Micropropagation and production of phytopharmaceutical plants |
| CN101869055A (en) * | 2009-04-23 | 2010-10-27 | 上海上房园林植物研究所 | Tissue culture method of red flower polygonum |
| CN104686341A (en) * | 2015-02-22 | 2015-06-10 | 梁仕华 | Tissue culture technique of aquilaria sinensis |
| CN107873524A (en) * | 2017-12-25 | 2018-04-06 | 绵阳市蜀创农业科技有限公司 | A kind of serpentgrass stem segment tissue culture fast breeding method |
Non-Patent Citations (1)
| Title |
|---|
| 刘静华: "火炭母组织培养灭菌方法探索", 《韶关学院学报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117426304A (en) * | 2023-12-20 | 2024-01-23 | 北京花乡花木集团有限公司 | Tissue culture method of polygonum giganteum |
| CN117426304B (en) * | 2023-12-20 | 2024-03-12 | 北京花乡花木集团有限公司 | Tissue culture method of polygonum giganteum |
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Application publication date: 20190118 |