CN106305420A - Medicinal wild rice embryo rescue rapid progeny propagation culture medium and method - Google Patents
Medicinal wild rice embryo rescue rapid progeny propagation culture medium and method Download PDFInfo
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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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Abstract
The invention provides a medicinal wild rice embryo rescue rapid progeny propagation culture medium. The medicinal wild rice embryo rescue rapid progeny propagation culture medium comprises an induction culture medium, a differentiation culture medium and a rooting culture medium, wherein the formula of the induction culture medium is MS+2.4D1.0-3.0mg/l+6BA0.1-0.5mg/l; the formula of the differentiation culture medium is MS+IAA1.0-3.0mg/l+6BA2.0-4.0mg/l+KT1.0-2.0mg/l; and the formula of the rooting culture medium is 1/2MS+IAA0.02-0.5mg/l. The invention also provides a medicinal wild rice embryo rescue rapid progeny propagation method. According to the medicinal wild rice embryo rescue rapid progeny propagation culture medium and method provided by the invention, healthy and rapid growth of tissue culture seedlings can be realized a short period of time.
Description
Technical field
The present invention relates to plant tissue culture fast breeding technique field, especially relate to a kind of oryza officinalis embryo rescue offspring quick
The culture medium of breeding and method.
Background technology
Wild rice is that the valuable of rice breeding plants matter.Breeding research both at home and abroad shows, has the genomic wild rice of AA type
Huge effect was played in rice breeding.Breeding men generally acknowledge, will obtain breakthrough breeding, it is necessary to find new close source.Can
See that to utilize wild rice to make parent imperative.But because cultivated rice is the tool genomic seed rice of AA type and common wild-rice hybridization appearance
Easily success, and hybridize with the non-genomic seed rice of AA type, often lead to failure by generally conventional sexual hybridization method.And it is out of office
The non-genomic wild rice of AA type in raw rice, the most often has the excellent character that cultivation is rare with Oryza rufipogon.
Oryza officinalis is one of existing three kinds of wild rices of China, and early-stage Study result shows, oryza officinalis is to ash
Plant hopper has the strongest resistance and driving property of row.In order to make full use of oryza officinalis parent source, the researcher such as tight Cheng Qi is engaged in
Oryza officinalis for many years and the correlational study of cultivated rice embryo rescue, it is thus achieved that oryza officinalis embryo rescue offspring YF2, it is achieved
The hereditary material of oryza officinalis shifts to cultivated rice.
At present, by this technological means of tissue culture be solve oryza officinalis embryo rescue offspring's Fast-propagation effective
Measure, but the culture medium used often with the addition of substantial amounts of chemosynthesis phytohormone to promote micropropagation of plants and life
Long, in cultivating oryza officinalis embryo rescue offspring's reproductive process, often there is also tissue cultured seedling resistance based on this culture medium poor
And the low inferior defect of planting percent.
Therefore, it is necessary to provide a kind of novel culture medium, and on the basis of this culture medium, oryza officinalis embryo is saved
Rescue offspring's method for quickly breeding and produce improvement, to overcome drawbacks described above.
Summary of the invention
In order to solve above-mentioned prior art to cause oryza officinalis embryo save offspring's Fast-propagation because of medium component
During there is tissue cultured seedling resistance difference and the low technical problem of planting percent, the present invention provides the oryza officinalis of a kind of improvement
The culture medium of embryo rescue offspring's Fast-propagation and method, train with group during reaching oryza officinalis embryo rescue offspring's Fast-propagation
Seedling is healthy and the purpose of fast numerous growth.
The present invention provides the culture medium of a kind of oryza officinalis embryo rescue offspring's Fast-propagation, and described oryza officinalis embryo is saved
The culture medium rescuing offspring's Fast-propagation is made up of inducing culture, division culture medium and root media；
The formula of described inducing culture is: MS+2.4D 1.0-3.0mg/l+6BA0.1-0.5mg/l；
The formula of described division culture medium is: MS+IAA 1.0-3.0mg/l+6BA2.0-4.0mg/l+KT 1.0-
2.0mg/l；
The formula of described root media is: 1/2MS+IAA 0.02-0.5mg/l.
In culture medium one preferred embodiment of described oryza officinalis embryo rescue offspring's Fast-propagation of present invention offer,
The formula of described inducing culture is: MS+2.4D 2.0mg/l+6BA0.3mg/l, and the formula of described division culture medium is: MS+
IAA 1.8mg/l+6BA2.5mg/l+KT 1.2mg/l, the formula of described root media is: 1/2MS+IAA0.03mg/l.
In culture medium one preferred embodiment of described oryza officinalis embryo rescue offspring's Fast-propagation of present invention offer,
The pH value of described inducing culture, division culture medium and root media is 5.5-5.9.
The present invention also provides for a kind of oryza officinalis embryo rescue offspring's rapid propagation method, comprises the steps:
Inducing culture: be inoculated in above-mentioned inducing culture after the seed of oryza officinalis embryo rescue offspring is carried out pretreatment
On carry out inducing culture, induce cells,primordial；
Differentiation culture: cells,primordial is inoculated on above-mentioned division culture medium and carries out differentiation culture, differentiate seedling；
Root culture: be inoculated in by seedling on above-mentioned root media and carry out root culture, sends out roots and is tied to form as complete examination
Guan Miao.
In described oryza officinalis embryo rescue offspring's rapid propagation method one preferred embodiment that the present invention provides, institute
Stating pretreatment is: the seed of oryza officinalis embryo rescue offspring is put into sterilizing fine sand and carries out heat treatment 6-at 37 ± 2 DEG C
12 thoughtful seedling；Take plumelet 1-2cm, add one through 70% alcohol disinfecting 30 seconds, 0.1% mercuric chloride, shake and sterilize 8 minutes and aseptic
After washing 3-5 time, standby as outer implant.
In described oryza officinalis embryo rescue offspring's rapid propagation method one preferred embodiment that the present invention provides, institute
The condition of culture stating inducing culture is: temperature 25 ± 2 DEG C, light intensity 1000lux, illumination 16 hour/day, strengthens and arrive to surrounding
2000lux, strengthens after six weeks to 4000lux, cultivates 1.5-2 month to inducing cells,primordial.
In described oryza officinalis embryo rescue offspring's rapid propagation method one preferred embodiment that the present invention provides, institute
The condition of culture stating differentiation culture is: temperature 25 ± 2 DEG C, and light intensity 2000-3000lux, illumination 16 hour/day, 1 month to growing up to
Seedling.
In described oryza officinalis embryo rescue offspring's rapid propagation method one preferred embodiment that the present invention provides, institute
The condition of culture stating root culture is: temperature 25 ± 2 DEG C, light intensity 2000-3000lux, illumination 16 hour/day, sends out roots after 1 week
It is tied to form as complete test tube Seedling.
Compared to prior art, the culture medium of described oryza officinalis embryo rescue offspring's Fast-propagation that the present invention provides and
Method has the advantages that
One, inducing culture makes the callus cell quality of induction hard, faint yellow, to form cells,primordial fast；Differentiation
Culture medium makes from cells,primordial differentiation green bud point frequency high, has the 58% green bud of generation from 100 cells,primordials；Root culture
Base makes the green bud 90% of all inoculations take root and obtain green Seedling.
Two, oryza officinalis embryo rescue offspring the most mitogenetic go out substantial amounts of indefinite Multiple Buds, high-frequency regeneration plant, both side
The most efficient, cost-effective.
Three, the oryza officinalis embryo of rapid, high volume breeding in a short time rescue offspring's test tube Seedling, growth rate is fast, the production cycle
Short, breeding coefficient is high, can accomplish scale production.
Four, rapid, high volume breeds the tissue culture's test tube Seedling obtained, and heritability is consistent, overcomes conventional seminal propagation offspring
The easily shortcoming of variation；Additionally, use outer implant material few, can effectively save rare or endangered species, be conducive to protecting wild resource.
Accompanying drawing explanation
For the technical scheme being illustrated more clearly that in the embodiment of the present invention, in embodiment being described below required for make
Accompanying drawing be briefly described, it should be apparent that, below describe in accompanying drawing be only some embodiments of the present invention, for
From the point of view of those of ordinary skill in the art, on the premise of not paying creative work, it is also possible to obtain other according to these accompanying drawings
Accompanying drawing, wherein:
The medicine that the culture medium of oryza officinalis embryo rescue offspring's Fast-propagation that Fig. 1 provides by the present invention and method are cultivated
Tissue cultured seedling with wild rice embryo rescue offspring.
Detailed description of the invention
Below in conjunction with the accompanying drawing in the embodiment of the present invention, the technical scheme in the embodiment of the present invention is carried out clear, complete
Describe, it is clear that described embodiment is only a part of embodiment of the present invention rather than whole embodiments wholely.Based on
Embodiment in the present invention, those of ordinary skill in the art obtained under not making creative work premise all other
Embodiment, broadly falls into the scope of protection of the invention.
One of the material that the present invention uses oryza officinalis embryo rescue progeny seed YF2, is stored in Agriculture of Zhejiang Province science
Institute's virus and biotechnology research institute.Oryza officinalis embryo rescue offspring be oryza officinalis and cultivated rice protoplast asymmetric
The offspring that somatic embryo rescue produces, it has the strongest resistance and driving property of row to small brown rice planthopper.
The present invention provides the culture medium of a kind of oryza officinalis embryo rescue offspring's Fast-propagation, and described oryza officinalis embryo is saved
The culture medium rescuing offspring's Fast-propagation is made up of inducing culture, division culture medium and root media；
The formula of described inducing culture is: MS+2.4D 1.0-3.0mg/l+6BA0.1-0.5mg/l；
The formula of described division culture medium is: MS+IAA1.0-3.0mg/l+6BA2.0-4.0mg/l+KT 1.0-2.0mg/
l；
The formula of described root media is: 1/2MS+IAA0.02-0.5mg/l.
The pH value of described inducing culture, division culture medium and root media is 5.5-5.9.
Described MS is minimal medium；Described 2.4D is 2,4-dichlorphenoxyacetic acid, auxin, can promote thin in group training
The division of born of the same parents and growth；Described 6BA is 6 benzyladenines, and the basic element of cell division can promote cell division；Described IAA is indole second
Acid, as plant growth substance；Described KT is kinetins, is mainly used in tissue culture, promotes cell division and regulation cell
Differentiation.
The present invention also provides for a kind of method of culture medium based on described oryza officinalis embryo rescue offspring's Fast-propagation, bag
Include following steps:
Inducing culture: be inoculated in described inducing culture after the seed of oryza officinalis embryo rescue offspring is carried out pretreatment
On carry out inducing culture, induce cells,primordial；
Differentiation culture: cells,primordial is inoculated on described division culture medium and carries out differentiation culture, differentiate seedling；
Root culture: be inoculated in by seedling on described root media and carry out root culture, sends out roots and is tied to form as complete examination
Guan Miao.
Embodiment 1:
1) oryza officinalis embryo rescue progeny seed pretreatment: oryza officinalis embryo rescue progeny seed is put into sterilizing thin
At 37 ± 2 DEG C, the thoughtful seedling of heat treatment 6-12 is carried out in Sha；Take plumelet 1-2cm, through 70% alcohol disinfecting 30 seconds, 0.1% liter
Hydrargyrum add one, shake sterilization 8 minutes and aseptic washing 3-5 time after, standby as outer implant；
2) inducing culture: be seeded on inducing culture by seed, the formula of described inducing culture is MS+2.4D
2.0mg/l+6BA0.3mg/l；Condition of culture is at 25 ± 2 DEG C, light intensity 1000lux, illumination 16 hour/day, strengthens to surrounding
To 2000lux, strengthen after six weeks to 4000lux, cultivate 1.5-2 month to inducing cells,primordial；
3) differentiation culture: be seeded on division culture medium by cells,primordial, the formula of described division culture medium is MS+IAA
1.8mg/l+6BA 2.5mg/l+KT 1.2mg/l；Condition of culture is at 25 ± 2 DEG C, and light intensity 2000-3000lux, illumination 16 are little
Time/sky, 1 month to growing up to seedling；
4) root culture: seedling is inoculated on root media, described root media 1/2MS+IAA 0.03mg/l；
Condition of culture is at 25 ± 2 DEG C, light intensity 2000-3000lux, illumination 16 hour/day, sends out roots and be tied to form as complete test tube after 1 week
Seedling.Carry out the complete plant that regenerates high for plantlet of transplant: 4-6cm afterwards to transplant in rice soil.Above-mentioned each stage culture medium
PH value is 5.5.Tissue cultured seedling growth result sees Fig. 1.
Embodiment 2:
1) oryza officinalis embryo rescue progeny seed pretreatment: oryza officinalis embryo rescue progeny seed is put into sterilizing thin
At 37 ± 2 DEG C, the thoughtful seedling of heat treatment 6-12 is carried out in Sha；Take plumelet 1-2cm, through 70% alcohol disinfecting 30 seconds, 0.1% liter
Hydrargyrum add one, shake sterilization 8 minutes and aseptic washing 3-5 time after, standby as outer implant；
2) inducing culture: be seeded on inducing culture by seed, the formula of described inducing culture is S+2.4D
1.0mg/l+6BA 0.1mg/l；Condition of culture is at 25 ± 2 DEG C, light intensity 1000lux, illumination 16 hour/day, increases to surrounding
Strong to 2000lux, strengthen after six weeks to 4000lux, cultivate 1.5-2 month to inducing cells,primordial；
3) differentiation culture: be seeded on division culture medium by cells,primordial, the formula of described division culture medium is MS+IAA
1.0mg/l+6BA 2.0mg/l+KT 1.0mg/l；Condition of culture is at 25 ± 2 DEG C, and light intensity 2000-3000lux, illumination 16 are little
Time/sky, 1 month to growing up to seedling；
4) root culture: seedling is inoculated on root media, described root media 1/2MS+IAA 0.02mg/l；
Condition of culture is at 25 ± 2 DEG C, light intensity 2000-3000lux, illumination 16 hour/day, sends out roots and be tied to form as complete test tube after 1 week
Seedling.Carry out the complete plant that regenerates high for plantlet of transplant: 4-6cm afterwards to transplant in rice soil.Above-mentioned each stage culture medium
PH value is 5.5.
Embodiment 3:
1) oryza officinalis embryo rescue progeny seed pretreatment: oryza officinalis embryo rescue progeny seed is put into sterilizing thin
At 37 ± 2 DEG C, the thoughtful seedling of heat treatment 6-12 is carried out in Sha；Take plumelet 1-2cm, through 70% alcohol disinfecting 30 seconds, 0.1% liter
Hydrargyrum add one, shake sterilization 8 minutes and aseptic washing 3-5 time after, standby as outer implant；
2) inducing culture: be seeded on inducing culture by seed, the formula of described inducing culture is MS+2.4D
3.0mg/l+6BA0.5mg/l；Condition of culture is at 25 ± 2 DEG C, light intensity 1000lux, illumination 16 hour/day, strengthens to surrounding
To 2000lux, strengthen after six weeks to 4000lux, cultivate 1.5-2 month to inducing cells,primordial；
3) differentiation culture: be seeded on division culture medium by cells,primordial, the formula of described division culture medium is MS+IAA
3.0mg/l+6BA 4.0mg/l+KT 2.0mg/l；Condition of culture is at 25 ± 2 DEG C, and light intensity 2000-3000lux, illumination 16 are little
Time/sky, 1 month to growing up to seedling；
4) root culture: seedling is inoculated on root media, described root media 1/2MS+IAA 0.5mg/l；
Condition of culture is at 25 ± 2 DEG C, light intensity 2000-3000lux, illumination 16 hour/day, sends out roots and be tied to form as complete test tube after 1 week
Seedling.Carry out the complete plant that regenerates high for plantlet of transplant: 4-6cm afterwards to transplant in rice soil.Above-mentioned each stage culture medium
PH value is 5.9.
The described oryza officinalis embryo rescue culture medium of offspring's Fast-propagation that the present invention provides and method have and following have
Benefit effect:
One, inducing culture makes the callus cell quality of induction hard, faint yellow, to form cells,primordial fast；Differentiation
Culture medium makes from cells,primordial differentiation green bud point frequency high, has the 58% green bud of generation from 100 cells,primordials；Root culture
Base makes the green bud 90% of all inoculations take root and obtain green Seedling.
Two, oryza officinalis embryo rescue offspring the most mitogenetic go out substantial amounts of indefinite Multiple Buds, high-frequency regeneration plant, both side
The most efficient, cost-effective.
Three, the oryza officinalis embryo of rapid, high volume breeding in a short time rescue offspring's test tube Seedling, growth rate is fast, the production cycle
Short, breeding coefficient is high, can accomplish scale production.
Four, rapid, high volume breeds the tissue culture's test tube Seedling obtained, and heritability is consistent, overcomes conventional seminal propagation offspring
The easily shortcoming of variation；Additionally, use outer implant material few, can effectively save rare or endangered species, be conducive to protecting wild resource.
The foregoing is only embodiments of the invention, not thereby limit the scope of the claims of the present invention, every utilize this
Equivalent structure or equivalence flow process that bright description and accompanying drawing content are made convert, or are directly or indirectly used in other relevant skill
Art field, is the most in like manner included in the scope of patent protection of the present invention.
Claims (8)
1. the culture medium of oryza officinalis embryo rescue offspring's Fast-propagation, it is characterised in that described oryza officinalis embryo is saved
The culture medium rescuing offspring's Fast-propagation is made up of inducing culture, division culture medium and root media；Wherein,
The formula of described inducing culture is: MS+2.4D 1.0-3.0mg/l+6BA 0.1-0.5mg/l；
The formula of described division culture medium is: MS+IAA 1.0-3.0mg/l+6BA 2.0-4.0mg/l+KT 1.0-2.0mg/l；
The formula of described root media is: 1/2MS+IAA 0.02-0.5mg/l.
The culture medium of oryza officinalis embryo the most according to claim 1 rescue offspring's Fast-propagation, it is characterised in that described
The formula of inducing culture is: MS+2.4D 2.0mg/l+6BA 0.3mg/l, and the formula of described division culture medium is: MS+IAA
1.8mg/l+6BA 2.5mg/l+KT 1.2mg/l, the formula of described root media is: 1/2MS+IAA 0.03mg/l.
The culture medium of oryza officinalis embryo the most according to claim 1 rescue offspring's Fast-propagation, it is characterised in that described
The pH value of inducing culture, division culture medium and root media is 5.5-5.9.
4. oryza officinalis embryo rescue offspring's rapid propagation method, it is characterised in that comprise the steps:
Inducing culture: be inoculated in after the seed of oryza officinalis embryo rescue offspring is carried out pretreatment and lure as claimed in claim 1
Lead and in culture medium, carry out inducing culture, induce cells,primordial；
Differentiation culture: cells,primordial is inoculated on division culture medium as claimed in claim 1 and carries out differentiation culture, differentiate little
Seedling；
Root culture: seedling is inoculated on root media as claimed in claim 1 and carries out root culture, send out roots be tied to form into
Complete test tube Seedling.
Oryza officinalis embryo the most according to claim 4 rescue offspring's rapid propagation method, it is characterised in that described pre-
It is processed as: the seed of oryza officinalis embryo rescue offspring is put into sterilizing fine sand at 37 ± 2 DEG C, carries out heat treatment 6-12 week
To seedling；Take plumelet 1-2cm, add one, shake sterilization 8 minutes and aseptic washing through 70% alcohol disinfecting 30 seconds, 0.1% mercuric chloride
After 3-5 time, standby as outer implant.
Oryza officinalis embryo the most according to claim 5 rescue offspring's rapid propagation method, it is characterised in that described in lure
The condition of culture leading cultivation is: temperature 25 ± 2 DEG C, light intensity 1000lux, illumination 16 hour/day, strengthens and arrive to surrounding
2000lux, strengthens after six weeks to 4000lux, cultivates 1.5-2 month to inducing cells,primordial.
Oryza officinalis embryo the most according to claim 6 rescue offspring's rapid propagation method, it is characterised in that described point
Changing the condition of culture cultivated is: temperature 25 ± 2 DEG C, light intensity 2000-3000lux, illumination 16 hour/day, 1 month little to growing up to
Seedling.
Oryza officinalis embryo the most according to claim 7 rescue offspring's rapid propagation method, it is characterised in that described life
The condition of culture that root is cultivated is: temperature 25 ± 2 DEG C, light intensity 2000-3000lux, illumination 16 hour/day, sends out roots and be tied to form after 1 week
For complete test tube Seedling.
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| CN109089878A (en) * | 2018-07-18 | 2018-12-28 | 云南省农业科学院生物技术与种质资源研究所 | A method of regeneration plant is obtained using Oryza eichingeri bud for explant |
| CN110800734A (en) * | 2019-11-21 | 2020-02-18 | 上海市农业生物基因中心 | Ultra-low temperature preservation method for wild rice stem tips |
| CN112189557A (en) * | 2020-10-14 | 2021-01-08 | 浙江师范大学 | Breeding method of oversized panicle type rice based on morphological marker |
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| CN109089878A (en) * | 2018-07-18 | 2018-12-28 | 云南省农业科学院生物技术与种质资源研究所 | A method of regeneration plant is obtained using Oryza eichingeri bud for explant |
| CN110800734A (en) * | 2019-11-21 | 2020-02-18 | 上海市农业生物基因中心 | Ultra-low temperature preservation method for wild rice stem tips |
| CN112189557A (en) * | 2020-10-14 | 2021-01-08 | 浙江师范大学 | Breeding method of oversized panicle type rice based on morphological marker |
| CN112189557B (en) * | 2020-10-14 | 2021-10-19 | 浙江师范大学 | Breeding method of oversized panicle type rice based on morphological marker |
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| CN106305420B (en) | 2018-06-05 |
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