CN106069754A - A kind of Arundo donax tissue culture special culture media and cultural method - Google Patents
A kind of Arundo donax tissue culture special culture media and cultural method Download PDFInfo
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- CN106069754A CN106069754A CN201610429725.1A CN201610429725A CN106069754A CN 106069754 A CN106069754 A CN 106069754A CN 201610429725 A CN201610429725 A CN 201610429725A CN 106069754 A CN106069754 A CN 106069754A
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- arundo donax
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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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- 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
- A01H4/001—Culture apparatus for tissue culture
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- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
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- Botany (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention provides a kind of Arundo donax tissue culture special culture media, comprise component and content is as follows: MS minimal medium, agar 6g/l, indolebutyric acid (IBA) 0.5 1mg/l, activated carbon 0.1 0.5mg/l, Thidiazuron 0.5 1mg/l.During Arundo donax tissue culture propagation, the application with the addition of the TDZ of low dosage the most in the medium, in the enrichment culture stage, can produce substantial amounts of bud simultaneously, and increasing numerous amount is 1.3 1.5 times during the culture medium without TDZ.
Description
Technical field
The invention belongs to field of plant tissue culture technique, relate generally to a kind of Arundo donax tissue culture procedures improves breeding
Efficiency special culture media and cultural method.
Background technology
At present, Arundo donax (Arundo donax) is that grass family Arundo donax belongs to perennial tall and big tussock plant, has prosperity
Root stock.It is adaptable, has certain endurance to saline and alkaline, arid, barren, is referred to as the " first of saline-alkali wetland
Cutting edge of a knife or a sword plant ".Arundo donax not only has preventing and treating soil degradation and increases soil carbon sequestration and the ecological benefits of organic matter, wet for repairing
Ground heavy metal pollution also has bigger potentiality.In recent years, Arundo donax is also gradually being subject to as the value of the clean biometric energy
The attention of people, perennial herb lignocellulosic plants is considered to best suit biomass bioenergy and produces.
Owing to Arundo donax can not lean on seed growing, traditional tillering propagation, far from meeting gardens demand and ecological demand.Cause
This, utilize plant tissue culture technique by research and development special culture media and cultural method, accelerate the breeding cycle of Arundo donax, improve numerous
Growing efficiency, being that Arundo donax is large-scale industrialized produces key.
Summary of the invention
For solving the problem that reproductive efficiency is low present in above-mentioned background technology, the invention provides a kind of Arundo donax tissue and train
Reproductive efficiency special culture media and the cultural method of Arundo donax thereof is improved during Yanging.
It is an object of the invention to be achieved through the following technical solutions:
A kind of Arundo donax tissue culture special culture media, comprises component and content is as follows:
MS minimal medium, agar 6g/l, indolebutyric acid (IBA) 0.5-1mg/l, activated carbon 0.1-0.5mg/l, Thidiazuron
(TDZ)0.5-1mg/l。
The cultural method of a kind of Arundo donax tissue utilizing above-mentioned special culture media, comprises the steps:
(1) selection of outer implant and sterilization
Choose Arundo donax axillalry bud as outer implant, after rinsing 1.5-3 hour under flowing water, move in superclean bench, at 70-
The ethanol of 75% soaks the 20-60 second, moves into and shake 5-7 minute equipped with in the ultrasonic washing unit of liquor natrii hypochloritis, then use
Aseptic water washing 2-5 time；
(2) inoculation and the cultivation of aseptic strain
Aseptically, cut 0.5-2cm stem with bud, be inoculated into equipped with in the culture bottle of special culture media, 13-15
After it, it is thus achieved that aseptic strain；
(3) switching and expanding propagation
Aseptic bud in the aseptic strain of step (2) is aseptically stripped the group of 5-10 mm band apical growing point
Knit, be seeded on special culture media, carry out enrichment culture, 3-4 week repeat subculture once；
(4) root culture and domestication
Take the aseptic bud 2-3 strain obtained in step (3) to be one group and move in root media, will after continuing to cultivate 2-3 week
Plant is taken out after cleaning use carbendazim diluent immersion 5-20 minute, transplants to seedling medium, uses 50% sunshade afterwards
The sunshade net shading of rate 7-10 days.
Preferably, in described step (1), the concentration of liquor natrii hypochloritis is 5-10%.
Preferably, the concentration of described liquor natrii hypochloritis is 8%.
Preferably, the aseptic strain condition of culture in described step (2): intensity of illumination is 2500-3000lus, temperature is white
They 25 DEG C, night 16 DEG C, light application time is 11 hours/day.
Preferably, described special culture media is the Arundo donax tissue culture special culture media described in claim 1.
Preferably, in described step (3), the container of special culture media is the airtight culture bag of 10*13cm.
Preferably, in described step (4), the container of root media is the airtight culture bag of 10*13cm.
Preferably, the concentration of the carbendazim diluent in described step (4) is 0.1-0.3%.
Preferably, in described step (4), root culture based component is MS minimal medium, 0.1-0.3mg/l naphthalene acetic acid
(NAA), 0.3-0.8mg/l activated carbon.
Beneficial effects of the present invention:
1, during Arundo donax tissue culture propagation, with the addition of the TDZ of low dosage the most in the medium, Arundo donax bud is sprouted tool
There is obvious facilitation；In the enrichment culture stage, can produce substantial amounts of bud, increasing numerous amount is the culture medium without TDZ simultaneously
Time 1.3-1.5 times；TDZ concentration is sprouted at 0.5-1mg/l Arundo donax adventitious bud and is had obvious facilitation.After inoculation 7-10 days
Culture medium there is adventitious bud to occur with regard to the visible base portion at inoculation bud, inducing clumping bud can be formed 3-4 week and generate, subculture cycle
Shorten 1-2 week.TDZ concentration is less than 0.5mg/l, and outer planting body shows as the increase of height and the growth of blade, and the most a small amount of bud is sent out
Raw.TDZ concentration then starts tissue vitrification phenomenon (the i.e. leaf of seedling, tender tip water logging occur higher than the seedling in 1mg/l culture medium
Shape, in crystal shaped translucent, plant is short and small, chlorosis, and leaf-shrinkage crimps, frangible).
2, in the enrichment culture stage, use 10*13cm culture bag as culture vessel, subculture cycle was shortened from 4-5 week
To 3-4 week, accelerate reproduction speed；
3, the present invention can significantly improve the reproductive efficiency of Arundo donax, trains cost for reduction group, raw for Arundo donax batch production scale
Product has established solid foundation.
Detailed description of the invention
In order to the present invention is better described, below in conjunction with the embodiment of the present invention technical scheme carried out clear,
It is fully described by.
Embodiment 1
A kind of Arundo donax tissue culture special culture media, comprises component and content is as follows: MS minimal medium, agar 6g/l,
Indolebutyric acid (IBA) 0.8mg/l, activated carbon 0.3mg/l；Thidiazuron: 1mg/l.
Improving the cultural method of reproductive efficiency in a kind of Arundo donax tissue culture procedures, step is as follows:
1, the selection of outer implant and sterilization
Choose the axillalry bud of full disease-free Arundo donax as outer implant, after rinsing 2 hours under flowing water, move into ultra-clean work
In platform, soak 30 seconds in the ethanol of 75%, move into and shake 6 points equipped with in the ultrasonic washing unit of the liquor natrii hypochloritis of 8%
Clock, then with aseptic water washing 4 times；
2, inoculation and the foundation of aseptic strain
Aseptically, cut 1cm stem with bud, be inoculated in the culture bottle equipped with above-mentioned special culture media.2 weeks
After, stem Duan Shangya point sprouting is expanded, it is thus achieved that aseptic strain；
3, switching and expanding propagation
By above-mentioned steps obtain aseptic strain in aseptic bud aseptically strip 6 mm band apical growing points
Tissue, is seeded on above-mentioned special culture media, and culture vessel is replaced by 10*13cm culture bag, and culture bag is airtight, breeds
Cultivating, 3 weeks repeat subculture once；
4, root culture and domestication
Above-mentioned steps obtaining aseptic bud 2-3 strain be one group and go in root media, container of taking root is that 10*13cm cultivates
Bag, takes out plant after continuing to cultivate 2 weeks and cleans culture medium, after using 0.1% carbendazim to soak 10 minutes, transplant to nursery base
In matter.Use the sunshade net shading 10 days of 50% sunshade rate afterwards.Domestication process is completed after transplanting 45 days.The one-tenth of root media
It is divided into MS minimal medium, 0.1mg/l naphthalene acetic acid (NAA), 0.5mg/l activated carbon.
Embodiment 2
A kind of Arundo donax tissue culture special culture media, comprises component and content is as follows: MS minimal medium, agar 6g/l,
Indolebutyric acid (IBA) 1mg/l, activated carbon 0.1mg/l；Thidiazuron: 0.8mg/l.
Improving the cultural method of reproductive efficiency in a kind of Arundo donax tissue culture procedures, step is as follows:
1, the selection of outer implant and sterilization
Choose the axillalry bud of full disease-free Arundo donax as outer implant, after rinsing 2.2 hours under flowing water, move into ultra-clean work
In station, soak 40 seconds in the ethanol of 73%, move into equipped with the ultrasonic washing unit of the liquor natrii hypochloritis of 8% shakes 5
Minute, then with aseptic water washing 5 times；
2, inoculation and the foundation of aseptic strain
Aseptically, cut 2cm stem with bud, be inoculated in the culture bottle equipped with above-mentioned special culture media.13 days
After, stem Duan Shangya point sprouting is expanded, it is thus achieved that aseptic strain；
3, switching and expanding propagation
By above-mentioned steps obtain aseptic strain in aseptic bud aseptically strip 5 mm band apical growing points
Tissue, is seeded on above-mentioned special culture media, and culture vessel is replaced by 10*13cm culture bag, and culture bag is airtight, breeds
Cultivating, 4 weeks repeat subculture once；
4, root culture and domestication
Above-mentioned steps obtaining aseptic bud 2-3 strain be one group and go in root media, container of taking root is that 10*13cm cultivates
Bag, takes out plant after continuing to cultivate 2 weeks and cleans culture medium, after using 0.1% carbendazim to soak 15 minutes, transplant to nursery base
In matter.Use the sunshade net shading 10 days of 50% sunshade rate afterwards.Domestication process is completed after transplanting 45 days.The one-tenth of root media
It is divided into MS minimal medium, 0.1mg/l naphthalene acetic acid (NAA), 0.5mg/l activated carbon.
The embodiment of the present application 1-2 and the contrast test statistical table of ordinary culture medium
The above, the only present invention preferably detailed description of the invention, but protection scope of the present invention is not limited thereto,
Any those familiar with the art in the technical scope of present disclosure, the change that can readily occur in or replacement,
All should contain within protection scope of the present invention.Therefore, protection scope of the present invention should be with the protection of claims
Scope is as the criterion.
Claims (10)
1. an Arundo donax tissue culture special culture media, it is characterised in that comprise component and content is as follows:
MS minimal medium, agar 6g/l, indolebutyric acid (IBA) 0.5-1mg/l, activated carbon 0.1-0.5mg/l, Thidiazuron
(TDZ)0.5-1mg/l。
2. the cultural method of the Arundo donax tissue utilizing special culture media described in claim 1, it is characterised in that include as follows
Step:
(1) selection of outer implant and sterilization
Choose Arundo donax axillalry bud as outer implant, after rinsing 1.5-3 hour under flowing water, move in superclean bench, at 70-75%
Ethanol in soak the 20-60 second, move into and shake 5-7 minute equipped with in the ultrasonic washing unit of liquor natrii hypochloritis, then with aseptic
Water rinses 2-5 time；
(2) inoculation and the cultivation of aseptic strain
Aseptically, cut 0.5-2cm stem with bud, be inoculated into equipped with in the culture bottle of special culture media, 13-15 days
After, it is thus achieved that aseptic strain；
(3) switching and expanding propagation
Aseptic bud in the aseptic strain of step (2) is aseptically stripped the tissue of 5-10 mm band apical growing point, connects
Plant on special culture media, carry out enrichment culture, 3-4 week repeat subculture once；
(4) root culture and domestication
Take the aseptic bud 2-3 strain obtained in step (3) to be one group and move in root media, continue to cultivate after 2-3 week by plant
Take out after cleaning use carbendazim diluent immersion 5-20 minute, transplant to seedling medium, use 50% sunshade rate afterwards
Sunshade net shading 7-10 days.
The cultural method of Arundo donax tissue the most according to claim 2, it is characterised in that sodium hypochlorite in described step (1)
The concentration of solution is 5-10%.
The cultural method of Arundo donax tissue the most according to claim 3, it is characterised in that the concentration of described liquor natrii hypochloritis
It is 8%.
The cultural method of Arundo donax tissue the most according to claim 2, it is characterised in that the aseptic strain in described step (2)
Condition of culture: intensity of illumination is 2500-3000lus, temperature is daytime 25 DEG C, and night 16 DEG C, light application time is 11 hours/day.
The cultural method of Arundo donax tissue the most according to claim 2, it is characterised in that described special culture media is that right is wanted
Seek the Arundo donax tissue culture special culture media described in 1.
The cultural method of Arundo donax tissue the most according to claim 2, it is characterised in that special cultivation in described step (3)
The container of base is the airtight culture bag of 10*13cm.
The cultural method of Arundo donax tissue the most according to claim 2, it is characterised in that root culture in described step (4)
The container of base is the airtight culture bag of 10*13cm.
The cultural method of Arundo donax tissue the most according to claim 2, it is characterised in that the carbendazim in described step (4)
The concentration of diluent is 0.1-0.3%.
The cultural method of Arundo donax tissue the most according to claim 2, it is characterised in that root culture in described step (4)
Based component is MS minimal medium, 0.1-0.3mg/l naphthalene acetic acid (NAA), 0.3-0.8mg/l activated carbon.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201610429725.1A CN106069754A (en) | 2016-06-16 | 2016-06-16 | A kind of Arundo donax tissue culture special culture media and cultural method |
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| CN201610429725.1A CN106069754A (en) | 2016-06-16 | 2016-06-16 | A kind of Arundo donax tissue culture special culture media and cultural method |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108029554A (en) * | 2017-12-04 | 2018-05-15 | 中国林业科学研究院热带林业研究所 | The sterilization method of Michelia macclurei explant in a kind of complete stand |
| CN110122334A (en) * | 2019-06-20 | 2019-08-16 | 运城学院 | A kind of giantreed quickly breeds special culture media and its cultural method |
| CN114208679A (en) * | 2022-01-19 | 2022-03-22 | 内蒙古农业大学 | Tissue culture rapid propagation method of oasis 1 juncao |
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| WO2002063023A2 (en) * | 2001-02-05 | 2002-08-15 | The University Of South Carolina Research Foundation | Sustained totipotent regenerable tissue culture of arundo donax (giant reed) and totipotent tissue and plants produced therefrom |
| US7052912B1 (en) * | 2001-01-04 | 2006-05-30 | Woods Susan H | Methods and apparatus for the micro- and macropropagation of reed grasses |
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| CN103229720A (en) * | 2013-04-26 | 2013-08-07 | 湖州师范学院 | Method for regenerating plant from arundo donax linn callus |
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| US20160050866A1 (en) * | 2014-08-20 | 2016-02-25 | The Institute For Advanced Learning And Research | Micropropagation and plant regeneration systems for arundo donax and other monocots |
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| US7052912B1 (en) * | 2001-01-04 | 2006-05-30 | Woods Susan H | Methods and apparatus for the micro- and macropropagation of reed grasses |
| WO2002063023A2 (en) * | 2001-02-05 | 2002-08-15 | The University Of South Carolina Research Foundation | Sustained totipotent regenerable tissue culture of arundo donax (giant reed) and totipotent tissue and plants produced therefrom |
| CN102919127A (en) * | 2012-11-12 | 2013-02-13 | 湖州师范学院 | Method for building bamboo reed tissue culture system |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108029554A (en) * | 2017-12-04 | 2018-05-15 | 中国林业科学研究院热带林业研究所 | The sterilization method of Michelia macclurei explant in a kind of complete stand |
| CN110122334A (en) * | 2019-06-20 | 2019-08-16 | 运城学院 | A kind of giantreed quickly breeds special culture media and its cultural method |
| CN110122334B (en) * | 2019-06-20 | 2023-03-28 | 运城学院 | Special culture medium for rapid propagation of arundo donax linn and culture method thereof |
| CN114208679A (en) * | 2022-01-19 | 2022-03-22 | 内蒙古农业大学 | Tissue culture rapid propagation method of oasis 1 juncao |
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Application publication date: 20161109 |