CN105850744A - Open tissue culture medium and preparation method thereof for eustoma russellianum - Google Patents
Open tissue culture medium and preparation method thereof for eustoma russellianum Download PDFInfo
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- CN105850744A CN105850744A CN201610309908.XA CN201610309908A CN105850744A CN 105850744 A CN105850744 A CN 105850744A CN 201610309908 A CN201610309908 A CN 201610309908A CN 105850744 A CN105850744 A CN 105850744A
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- culture medium
- crude extract
- tissue culture
- lisianthus
- garlic
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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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- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Developmental Biology & Embryology (AREA)
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Abstract
The invention provides open tissue culture medium and a preparation method thereof for eustoma russellianum. The culture medium comprises 25-35 g of cane sugar, 6-12 g of agar, 80-120 ml of crude garlic extract, 4090 mg of macroelements, 440 mg of Cacl2.2H2O, 38.23 mg of microelement, 1 mg of cytokinin, 65.1 mg of molysite, 3.1 mg of organic matters, and 100 mg of creatine by weight of 1 L. The culture medium has no need high-pressure sterilization during use and strict sterile operation as well, so that plants can growth well in the open condition, cost is greatly lowered and application and popularization of the plant tissue culture technology are benefited.
Description
Technical field
The present invention relates to tissue culture technology field, particularly relate to a kind of Lisianthus open tissue culture medium and preparation side thereof
Method.
Background technology
Owing to traditional Plant Tissue Breeding needs to operate under the conditions of strict gnotobasis, condition of work is wanted
Asking harsh, operation is not easy to control, and needs to put into substantial amounts of human and material resources, and tradition plantlet in vitro exists poor growth, pollutes tight
Weight, the problems such as survival rate is low so that Plant Tissue Breeding is difficult to be used widely and promote in actual industrial and agricultural production.
It is then that the culture medium by special configuration makes group cultivation thing grow under antibacterial environment that the open tissue risen at present is cultivated,
Plant is made can well to grow under conditions of opposing open.Cost can either be reduced, be beneficial to plant tissue culture technique again
Applying and be generalized in industrial seedling rearing production practices, the present invention is i.e. that open group of the Lisianthus that is specifically designed for of exploitation trains liquid.
Summary of the invention
In view of the deficiencies in the prior art, the invention reside in a kind of Lisianthus open tissue culture medium of offer and preparation side thereof
Method, in order to realize the method for culturing open type tissue of Lisianthus.
Technical scheme is as follows:
A kind of Lisianthus open tissue culture medium, wherein, based on 1L culture medium weight, described culture medium includes following components:
Sucrose 25-35g；
Agar 6-12g；
Garlic crude extract 80-120 ml；
Basic element of cell division 1mg；
Methyl α-naphthyl acetate 0.5mg；
KNO31900mg；
NH4NO31650mg；
MgSO4·7H2O 370mg；
KH2PO4170mg；
CaCl2·2H2O 440mg；
MnSO4·4H2O 22.3mg；
ZnSO4·7H2O 8.6mg；
H3BO36.2mg；
KI 0.83mg；
Na2MoO4·7H2O 0.25mg；
CoCL2·6H2O 0.025mg；
CuSO4·5H2O 0.025mg；
Molysite 65.1mg；
Organic matter 3.1mg；
Creatine 100mg；
Wherein, garlic crude extract concentration is 3g/ml.
Described Lisianthus open tissue culture medium, wherein, based on 1L culture medium weight, described culture medium includes following
Component:
Sucrose 30g；
Agar 9g；
Garlic crude extract 100 ml；
Basic element of cell division 1mg；
Methyl α-naphthyl acetate 0.5mg；
KNO31900mg；
NH4NO31650mg；
MgSO4·7H2O 370mg；
KH2PO4170mg；
CaCl2·2H2O 440mg；
MnSO4·4H2O 22.3mg；
ZnSO4·7H2O 8.6mg；
H3BO36.2mg；
KI 0.83mg；
Na2MoO4·7H2O 0.25mg；
CoCL2·6H2O 0.025mg；
CuSO4·5H2O 0.025mg；
Molysite 65.1mg；
Organic matter 3.1mg；
Creatine 100mg;
Wherein, garlic crude extract concentration is 3g/ml.
Described Lisianthus open tissue culture medium, wherein, based on 1L culture medium weight, described molysite is by following components
Composition:
Na2-EDTA 37.3mg；
FeSO4.4H2O 27.8mg。
Described Lisianthus open tissue culture medium, wherein, based on 1L culture medium weight, described organic matter is by following group
It is grouped into:
Glycine 2mg；
Puridoxine hydrochloride VB60.5mg；
Tyiamine Hd element VB10.1mg；
Nicotinic acid 0.5mg.
Described Lisianthus open tissue culture medium, wherein, described garlic crude extract is prepared by following process: remove
Skin garlic, adds appropriate 70% ethanol after mincing, squeeze garlic juice with juice extractor, then by 2 layers of filtered through gauze, take filtrate, by filtrate
By 4000r/min centrifuge 8min, taking supernatant, be settled to 100ml with 70% ethanol, the garlic obtaining 3.0g/mL is thick
Extract, in 4 DEG C of preservations.
The preparation method of a kind of Lisianthus open tissue culture medium as above, wherein, described method is: will be except sugarcane
Component outside sugar, agar, garlic crude extract weighs according to intended component and mixes, use 0.5mol/L hydrochloric acid solution and
Liquid medium is adjusted PH5.8 ~ 5.9 by 0.5mol/L sodium hydroxide solution, weigh in proportion addition sucrose, agar after heating boil melt mixed
Even, when culture medium is cooled to 50 DEG C, add garlic crude extract, prepare required culture medium.
Beneficial effect: the present invention provides a kind of Lisianthus open tissue culture medium and preparation method thereof, and this culture medium makes
Used time, without autoclaving, can realize plant without strict sterile working simultaneously and well give birth under conditions of opposing open
Long, it is substantially reduced cost, beneficially by application and the popularization of plant tissue culture technique.
Detailed description of the invention
The present invention provides a kind of Lisianthus open tissue culture medium and preparation method thereof, for making the purpose of the present invention, skill
Art scheme and effect are clearer, clear and definite, and the present invention is described in more detail below.Should be appreciated that tool described herein
Body embodiment only in order to explain the present invention, is not intended to limit the present invention.
The Lisianthus open tissue culture medium of the present invention, wherein, based on 1L culture medium weight, described culture medium include with
Lower component:
Sucrose 25-35g；
Agar 6-12g；
Garlic crude extract 80-120 ml；
Basic element of cell division 1mg；
Methyl α-naphthyl acetate 0.5mg；
KNO31900mg；
NH4NO31650mg；
MgSO4·7H2O 370mg；
KH2PO4170mg；
CaCl2·2H2O 440g；
MnSO4·4H2O 22.3mg；
ZnSO4·7H2O 8.6mg；
H3BO36.2mg；
KI 0.83mg；
Na2MoO4·7H2O 0.25mg；
CoCL2·6H2O 0.025mg；
CuSO4·5H2O 0.025mg；
Molysite 65.1mg；
Organic matter 3.1mg；
Creatine 100mg；
Wherein, garlic crude extract concentration is 3g/ml.
In preferred embodiment, based on 1L culture medium weight, described culture medium includes following components:
Sucrose 30g；
Agar 9g；
Garlic crude extract 100 ml；
Basic element of cell division 1mg；
Methyl α-naphthyl acetate 0.5mg；
KNO31900mg；
NH4NO31650mg；
MgSO4·7H2O 370mg；
KH2PO4170mg；
CaCl2·2H2O 440g；
MnSO4·4H2O 22.3mg；
ZnSO4·7H2O 8.6mg；
H3BO36.2mg；
KI 0.83mg；
Na2MoO4·7H2O 0.25mg；
CoCL2·6H2O 0.025mg；
CuSO4·5H2O 0.025mg；
Molysite 65.1mg；
Organic matter 3.1mg；
Creatine 100mg;
Wherein, garlic crude extract concentration is 3g/ml.And described molysite is composed of the following components:
Na2-EDTA 37.3mg；
FeSO4·4H2O 27.8mg。
Further, described organic matter is composed of the following components:
Glycine 2mg；
Puridoxine hydrochloride VB60.5mg；
Tyiamine Hd element VB10.1mg；
Nicotinic acid 0.5mg.
In specific embodiment, described garlic crude extract is prepared by following process: take peeling garlic, adds appropriate after mincing
70% ethanol, squeezes garlic juice with juice extractor, then by 2 layers of filtered through gauze, takes filtrate, by filtrate by 4000r/min centrifuge
Centrifugal 8min, takes supernatant, is settled to 100ml with 70% ethanol, obtain the garlic crude extract of 3.0g/mL, in 4 DEG C of preservations.
The present invention also provides for the preparation method of a kind of Lisianthus open tissue culture medium as above, wherein, described
Method is: the component in addition to sucrose, agar, garlic crude extract is weighed according to intended component and mixes, and uses 0.5mol/L
Liquid medium is adjusted PH5.8 ~ 5.9 by hydrochloric acid solution and 0.5mol/L sodium hydroxide solution, after weighing addition sucrose, agar in proportion
Heating is boiled and is melted mixing, adds garlic crude extract when culture medium is cooled to 50 DEG C, prepares required culture medium.
The present invention provides a kind of Lisianthus open tissue culture medium and preparation method thereof, without height when this culture medium uses
Pressure sterilizing, can realize plant without strict sterile working simultaneously and well grow under conditions of opposing open, significantly drop
Low cost, beneficially by application and the popularization of plant tissue culture technique.
It should be appreciated that the application of the present invention is not limited to above-mentioned citing, for those of ordinary skills, can
To be improved according to the above description or to convert, all these modifications and variations all should belong to the guarantor of claims of the present invention
Protect scope.
Claims (6)
1. a Lisianthus open tissue culture medium, it is characterised in that based on 1L culture medium weight, described culture medium include with
Lower component:
Sucrose 25-35g；
Agar 6-12g；
Garlic crude extract 80-120 ml；
Basic element of cell division 1mg；
Methyl α-naphthyl acetate 0.5mg；
KNO31900mg；
NH4NO31650mg；
MgSO4·7H2O 370mg；
KH2PO4170mg；
CaCl2·2H2O 440mg；
MnSO4·4H2O 22.3mg；
ZnSO4·7H2O 8.6mg；
H3BO36.2mg；
KI 0.83mg；
Na2MoO4·7H2O 0.25mg；
CoCL2·6H2O 0.025mg；
CuSO4·5H2O 0.025mg；
Molysite 65.1mg；
Organic matter 3.1mg；
Creatine 100mg；
Wherein, garlic crude extract concentration is 3g/ml.
Lisianthus open tissue culture medium the most according to claim 1, it is characterised in that based on 1L culture medium weight,
Described culture medium includes following components:
Sucrose 30g；
Agar 9g；
Garlic crude extract 100 ml；
Basic element of cell division 1mg；
Methyl α-naphthyl acetate 0.5mg；
KNO31900mg；
NH4NO31650mg；
MgSO4·7H2O 370mg；
KH2PO4170mg；
CaCl2·2H2O 440mg；
MnSO4·4H2O 22.3mg；
ZnSO4·7H2O 8.6mg；
H3BO36.2mg；
KI 0.83mg；
Na2MoO4·7H2O 0.25mg；
CoCL2·6H2O 0.025mg；
CuSO4·5H2O 0.025mg；
Molysite 65.1mg；
Organic matter 3.1mg；
Creatine 100mg;
Wherein, garlic crude extract concentration is 3g/ml.
Lisianthus open tissue culture medium the most according to claim 2, it is characterised in that based on 1L culture medium weight,
Described molysite is composed of the following components:
Na2-EDTA 37.3mg；
FeSO4·4H2O 27.8mg。
Lisianthus open tissue culture medium the most according to claim 2, it is characterised in that based on 1L culture medium weight,
Described organic matter is composed of the following components:
Glycine 2mg；
Puridoxine hydrochloride VB60.5mg；
Tyiamine Hd element VB10.1mg；
Nicotinic acid 0.5mg.
Lisianthus open tissue culture medium the most according to claim 1, it is characterised in that described garlic crude extract passes through
Prepared by following process: take peeling garlic, add appropriate 70% ethanol after mincing, and squeezes garlic juice with juice extractor, then by 2 layers of gauze
Filter, take filtrate, by filtrate by 4000r/min centrifuge 8min, take supernatant, be settled to 100ml with 70% ethanol,
Obtain the garlic crude extract of 3.0g/mL, in 4 DEG C of preservations.
6. a preparation method for the Lisianthus open tissue culture medium as described in any one of claim 1-5, its feature exists
In, described method is: the component in addition to sucrose, agar, garlic crude extract is weighed according to intended component and mixes, and uses
Liquid medium is adjusted PH5.8 ~ 5.9 by 0.5mol/L hydrochloric acid solution and 0.5mol/L sodium hydroxide solution, weighs addition sugarcane in proportion
After sugar, agar, heating is boiled and is melted mixing, adds garlic crude extract when culture medium is cooled to 50 DEG C, prepares required culture medium.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108575739A (en) * | 2018-01-23 | 2018-09-28 | 昆明学院 | The bacteria inhibiting composition and preparation method thereof of blueberry open tissue culture |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108575739A (en) * | 2018-01-23 | 2018-09-28 | 昆明学院 | The bacteria inhibiting composition and preparation method thereof of blueberry open tissue culture |
| CN108575739B (en) * | 2018-01-23 | 2021-08-03 | 昆明学院 | Bacteriostatic composition for open tissue culture of blueberries and preparation method thereof |
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Application publication date: 20160817 |