CN110463606A - A kind of taipei fritillary bulb bulb regenerating system method established using seed as explant - Google Patents
A kind of taipei fritillary bulb bulb regenerating system method established using seed as explant Download PDFInfo
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- CN110463606A CN110463606A CN201910837236.3A CN201910837236A CN110463606A CN 110463606 A CN110463606 A CN 110463606A CN 201910837236 A CN201910837236 A CN 201910837236A CN 110463606 A CN110463606 A CN 110463606A
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- seed
- bulb
- fritillary bulb
- taipei fritillary
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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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- 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)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The present invention relates to a kind of taipei fritillary bulb bulb regenerating system methods established using seed as explant, are related to plant regeneration field.The following steps are included: S1: obtaining taipei fritillary bulb seed；S2: the taipei fritillary bulb seed is pre-processed to obtain pretreatment seed；S3: the pretreatment seed is carried out disinfection to obtain disinfection seed；S4: Seed inducement callus is carried out to the disinfection seed and handles to obtain loose particles shape callus；S5: the loose particles shape callus is put into added with 6-BA and 2, is proliferated on the culture medium of 4-D, and continuous to be proliferated three generations and obtain a large amount of loose particles shape callus, wherein 30-40 days are a generation.This programme solves the technical issues of emergence rate for how improving good taipei fritillary bulb, suitable for cultivating the seedling of taipei fritillary bulb.
Description
Technical field
The present invention relates to plant regeneration field, in particular to a kind of taipei fritillary bulb bulb established using seed as explant is again
Raw body system, method.
Background technique
Taipei fritillary bulb (Fritillaria taipaiensis P.Y.Li) is the road place name in the Qin bar, Mount Taibai, mountain range area
Precious medicinal material is used as medicine with a long history, has the effect of moistening lung, resolving sputum, cough-relieving.Taipei fritillary bulb sexual propagation needs the 4-5 year could
It completes, and percentage of seedgermination is low, emergence difficult；Using asexual reproduction method, sowing quantity is big, and breeding coefficient is low, and demand is big, city
Supply falls short of demand always for field, causes production cost expensive.
Modern organization culture technique provides possibility for a large amount of quickly breeding plant seedlings.Have existing report, is all ether
The bulb of white fritillaria has the problems such as at high cost, pollution rate is high as explant.
Summary of the invention
The technical problem to be solved by the present invention is to how improve the emergence rate of good taipei fritillary bulb.
The technical scheme to solve the above technical problems is that a kind of taipei fritillary bulb established using seed as explant
Bulb regenerating system method, comprising the following steps:
S1: taipei fritillary bulb seed is obtained；
S2: the taipei fritillary bulb seed is pre-processed to obtain pretreatment seed；
S3: the pretreatment seed is carried out disinfection to obtain disinfection seed；
S4: Seed inducement callus is carried out to the disinfection seed and handles to obtain loose particles shape callus；
S5: the loose particles shape callus is put into added with 6-BA and 2, is proliferated on the culture medium of 4-D, continuous to be proliferated
Three generations obtains a large amount of loose particles shape callus, and wherein 30-40 days are a generation；
S6: the loose particles shape callus is inoculated in inductive differentiation medium and is broken up and obtained small squama
Stem, wherein placing 28 ± 1 DEG C of temperature, relative air humidity 70%-80% of the culturing room of inductive differentiation medium, light application time
12 hours/day, intensity of illumination 2000Lx, subculture is primary within 45 days；
S7: the clove is inoculated in root media and root induction obtains aseptic seedling of taking root, the culture of rootage
The cultivation temperature of culturing room where base is 28 ± 1 DEG C, relative air humidity 70%-80%, 12 hour/day of light application time, illumination
Intensity 2000Lx；
S8: the aseptic seedling of taking root is placed into culture bottle and is transferred in greenhouse to cultivate 1 week and is obtained initial stage rooted seedling；
S9: continuing to cultivate the initial stage rooted seedling 3-4 days rooted seedlings that strengthened, and continues to be gradually opened during culture
The bottleneck of the culture bottle；
S10: the root media is taken out from the culture bottle and cleaned to the reinforcement rooted seedling, by the reinforcement
Rooted seedling impregnates 10min in 1000 times of carbendazim solution and obtains transfer rooted seedling；
S11: the transplanting transfer rooted seedling is into matrix；
S12: every that the MS nutrient solution for applying addition NAA is poured after a predetermined time；
S13: film is covered in the matrix, by taking off film after a week.
The beneficial effects of the present invention are: this programme is handled by the seed to taipei fritillary bulb, pass through cleaning in the early stage
Disinfection reduces probability of the seed by pest and disease damage, and cultivates the callus of seed, obtains clove by callus, subtracts
The damage rate of fritillaria bulb during the growth process is lacked, clove is subjected to culture of rootage, compared to the bulb work with taipei fritillary bulb
For the scheme of explant, this programme reduces pollution rate and cost, how to improve going out for good taipei fritillary bulb to solve
The technical issues of seedling rate.
Based on the above technical solution, the present invention can also be improved as follows.
Further, step S2 is specifically included:
S21: clear water cleans the taipei fritillary bulb seed x hours, obtains clear water seed；
S22: the clear water seed is cleaned with the first predetermined solution, obtains the first seed；
S23: it is impregnated with the GA3 solution of 50-80mg/L and obtains pretreatment seed for 24 hours.
Further, step S3 is specifically included:
S31: the pretreatment seed 60-40s described in 75% alcohol solution dipping obtains ethyl alcohol taipei fritillary bulb seed；
S32: washing seed is obtained for ethyl alcohol taipei fritillary bulb seed 3 times described in sterile water wash；
S33: the washing seed is impregnated with 0.1% mercuric chloride solution；
S34: it is washed seed 5-6 times described in sterile water wash and obtains disinfection seed.
Further, step S4 is specifically included:
S41: the disinfection seed is seeded in the culture medium of Seed inducement healing cell group primary, the culture medium is put
It sets in culturing room, the cultivation temperature of the culturing room is 15 ± 1 DEG C；
S42: the culture medium is carried out shading treatment 30 days；
S43: lighting process is carried out 30-45 days to the culture medium and obtains loose particles shape callus, wherein illumination
Processing is intensity of illumination 2000Lx, 12 hour/day of light application time.
Beneficial effect using above-mentioned further scheme is, after disinfection seed inoculation in the medium, by training
The callus for supporting the change induction fritillaria of base illumination initially forms loose after lighting process at the kind hole of culture medium
Graininess callus.
Further, the matrix in step S11 is specially coco bran: vegetable garden soil: perlite: vermiculite=1:3:2:1.
The advantages of additional aspect of the invention, will be set forth in part in the description, and will partially become from the following description
It obtains obviously, or practice is recognized through the invention.
Detailed description of the invention
Fig. 1 is the method for the embodiment for the taipei fritillary bulb bulb regenerating system method that the present invention is established by explant of seed
Flow chart.
Specific embodiment
The principle and features of the present invention will be described below with reference to the accompanying drawings, and the given examples are served only to explain the present invention, and
It is non-to be used to limit the scope of the invention.
Embodiment is substantially as shown in Fig. 1:
The taipei fritillary bulb bulb regenerating system method established using seed as explant in the present embodiment, comprising:
The following steps are included:
S1: taipei fritillary bulb seed is obtained；
S2: taipei fritillary bulb seed is pre-processed to obtain pretreatment seed；
S3: pretreatment seed is carried out disinfection to obtain disinfection seed；
S4: carrying out Seed inducement callus to disinfection seed and handle to obtain loose particles shape callus, in the present embodiment
Disinfection seed is is seeded in seed germination medium by callus induction processing, and wherein the ingredient of seed germination medium includes MS
+ GA3 1.0~3.0mg/L+NAA, 0.01~0.2mg/L+ sugar 25g/L+ carragheen 8g/L；
S5: loose particles shape callus is put into added with 6-BA and 2, is proliferated on the proliferated culture medium of 4-D, the present embodiment
In the ingredient of proliferated culture medium include MS+6-BA 0.3~0.5mg/L+2,4-D, 0.2~0.6mg/L+ sugar 30g/L+ OK a karaoke club
Glue 8g/L, continuous proliferation three generations obtain a large amount of loose particles shape callus, and wherein 30-40 days are a generation；
S6: loose particles shape callus being inoculated in inductive differentiation medium and is broken up and obtained clove, this
The ingredient of inductive differentiation medium in embodiment includes MS+6-BA6.0~3.0mg/L+NAA 0.01~0.2mg/L+ sugar
25g/L+ carragheen 8g/L wherein places 28 ± 1 DEG C of temperature of the culturing room of inductive differentiation medium, relative air humidity 70%-
80%, 12 hour/day of light application time, intensity of illumination 2000Lx, subculture is primary within 45 days；
S7: inoculation clove is in root media and root induction obtains aseptic seedling of taking root, training where root media
The cultivation temperature for supporting room is 28 ± 1 DEG C, relative air humidity 70%-80%, 12 hour/day of light application time, intensity of illumination
2000Lx；
S8: the aseptic seedling that will take root, which is placed into culture bottle and is transferred in greenhouse to cultivate 1 week, obtains initial stage rooted seedling；
S9: continuing Initial stage of culture rooted seedling and strengthened within 3-4 days rooted seedling, continues to be gradually opened culture during cultivating
The bottleneck of bottle；
S10: will reinforce rooted seedling and root media taken out and cleaned from culture bottle, will reinforce rooted seedling at 1000 times
10min is impregnated in carbendazim solution obtains transfer rooted seedling；
S11: transplanting transfer rooted seedling is into matrix；
S12: every that the MS nutrient solution for applying addition NAA is poured after a predetermined time；
S13: covering film in matrix, by taking off film after a week.
The beneficial effects of the present invention are: this programme is handled by the seed to taipei fritillary bulb, pass through cleaning in the early stage
Disinfection reduces probability of the seed by pest and disease damage, and cultivates the callus of seed, obtains clove by callus, subtracts
The damage rate of fritillaria bulb during the growth process is lacked, clove is subjected to culture of rootage, compared to the bulb work with taipei fritillary bulb
For the scheme of explant, this programme reduces pollution rate and cost, how to improve going out for good taipei fritillary bulb to solve
The technical issues of seedling rate.
Based on the above technical solution, the present invention can also be improved as follows.
Optionally, in some other embodiments, step S2 is specifically included:
S21: clear water cleans taipei fritillary bulb seed x hours, obtains clear water seed；
S22: clear water seed is cleaned with the first predetermined solution, obtains the first seed；
S23: it is impregnated with the GA3 solution of 50-80mg/L and obtains pretreatment seed for 24 hours.
Optionally, in some other embodiments, step S3 is specifically included:
S31: ethyl alcohol taipei fritillary bulb seed is obtained with 75% alcohol solution dipping pretreatment seed 60-40s；
S32: washing seed is obtained with sterile water wash ethyl alcohol taipei fritillary bulb seed 3 times；
S33: with 0.1% mercuric chloride solution soaking and water washing seed, the mercury solution in the present embodiment is that 1g Hgcl2 is dissolved in
1000ml distilled water impregnates washing seed, concussion, and sterilize 12-15min, outwells mercury solution；
S34: it is washed seed 5-6 times with sterile water wash and obtains disinfection seed.
Optionally, in some other embodiments, step S4 is specifically included:
S41: disinfection seed is seeded in the culture medium of Seed inducement healing cell group primary, culture medium is placed on culture
In room, the cultivation temperature of culturing room is 15 ± 1 DEG C；
S42: culture medium is carried out shading treatment 30 days；
S43: lighting process is carried out 30-45 days to culture medium and obtains loose particles shape callus, wherein lighting process
For intensity of illumination 2000Lx, 12 hour/day of light application time.
After disinfection seed inoculation in the medium, pass through the callus group of the change induction fritillaria to culture medium illumination
It knits, after lighting process, loose particles shape callus is initially formed at the kind hole of culture medium.
Optionally, in some other embodiments, the matrix in step S11 is specially coco bran: vegetable garden soil: perlite: leech
Stone=1:3:2:1.
The advantages of additional aspect of the invention, will be set forth in part in the description, and will partially become from the following description
It obtains obviously, or practice is recognized through the invention.
It should be noted that the various embodiments described above are product embodiments corresponding with above-mentioned each method embodiment, for this
In embodiment the explanation of each constructional device and optional embodiment can with reference in above-mentioned each method embodiment pair it should be noted that In
This is repeated no more.
Reader should be understood that in the description of this specification reference term " one embodiment ", " is shown " some embodiments "
The description of example ", " specific example " or " some examples " etc. mean specific features described in conjunction with this embodiment or example, structure,
Material or feature are included at least one embodiment or example of the invention.In the present specification, above-mentioned term is shown
The statement of meaning property need not be directed to identical embodiment or example.Moreover, particular features, structures, materials, or characteristics described
It may be combined in any suitable manner in any one or more of the embodiments or examples.In addition, without conflicting with each other, this
The technical staff in field can be by the spy of different embodiments or examples described in this specification and different embodiments or examples
Sign is combined.
It is apparent to those skilled in the art that for convenience of description and succinctly, the dress of foregoing description
The specific work process with unit is set, can refer to corresponding processes in the foregoing method embodiment, details are not described herein.
More than, only a specific embodiment of the invention, but scope of protection of the present invention is not limited thereto, and it is any to be familiar with
Those skilled in the art in the technical scope disclosed by the present invention, can readily occur in various equivalent modifications or substitutions,
These modifications or substitutions should be covered by the protection scope of the present invention.Therefore, protection scope of the present invention should be wanted with right
Subject to the protection scope asked.
Claims (5)
1. a kind of taipei fritillary bulb bulb regenerating system method established using seed as explant, which is characterized in that including following step
It is rapid:
S1: taipei fritillary bulb seed is obtained；
S2: the taipei fritillary bulb seed is pre-processed to obtain pretreatment seed；
S3: the pretreatment seed is carried out disinfection to obtain disinfection seed；
S4: Seed inducement callus is carried out to the disinfection seed and handles to obtain loose particles shape callus；
S5: the loose particles shape callus is put into added with 6-BA and 2, is proliferated on the culture medium of 4-D, is continuously proliferated three generations
The a large amount of loose particles shape callus is obtained, wherein 30-40 days are a generation；
S6: the loose particles shape callus being inoculated in inductive differentiation medium and is broken up and obtained clove,
28 ± 1 DEG C of temperature, relative air humidity 70%-80% of the middle culturing room for placing inductive differentiation medium, light application time 12 is small
When/day, intensity of illumination 2000Lx, subculture is primary within 45 days；
S7: the clove is inoculated in root media and root induction obtains aseptic seedling of taking root, the root media institute
It is 28 ± 1 DEG C, relative air humidity 70%-80% in the cultivation temperature of culturing room, 12 hour/day of light application time, intensity of illumination
2000Lx；
S8: the aseptic seedling of taking root is placed into culture bottle and is transferred in greenhouse to cultivate 1 week and is obtained initial stage rooted seedling；
S9: continuing to cultivate the initial stage rooted seedling 3-4 days rooted seedlings that strengthened, and continues to be gradually opened during culture described
The bottleneck of culture bottle；
S10: taking out from the culture bottle and clean the root media for the reinforcement rooted seedling, and the reinforcement is taken root
Seedling impregnates 10min in 1000 times of carbendazim solution and obtains transfer rooted seedling；
S11: the transplanting transfer rooted seedling is into matrix；
S12: every that the MS nutrient solution for applying addition NAA is poured after a predetermined time；
S13: film is covered in the matrix, by taking off film after a week.
2. the taipei fritillary bulb bulb regenerating system method according to claim 1 established using seed as explant, feature
It is:
Step S2 is specifically included:
S21: clear water cleans the taipei fritillary bulb seed x hours, obtains clear water seed；
S22: the clear water seed is cleaned with the first predetermined solution, obtains the first seed；
S23: it is impregnated with the GA3 solution of 50-80mg/L and obtains pretreatment seed for 24 hours.
3. the taipei fritillary bulb bulb regenerating system method according to claim 1 established using seed as explant, feature
It is:
Step S3 is specifically included:
S31: the pretreatment seed 60-40s described in 75% alcohol solution dipping obtains ethyl alcohol taipei fritillary bulb seed；
S32: washing seed is obtained for ethyl alcohol taipei fritillary bulb seed 3 times described in sterile water wash；
S33: the washing seed is impregnated with 0.1% mercuric chloride solution；
S34: it is washed seed 5-6 times described in sterile water wash and obtains disinfection seed.
4. the taipei fritillary bulb bulb regenerating system method according to claim 1 established using seed as explant, feature
It is:
Step S4 is specifically included:
S41: the disinfection seed is seeded in the culture medium of Seed inducement healing cell group primary, the culture medium is placed on
In culturing room, the cultivation temperature of the culturing room is 15 ± 1 DEG C；
S42: the culture medium is carried out shading treatment 30 days；
S43: carrying out lighting process 30-45 days to the culture medium and obtain loose particles shape callus,
Wherein lighting process is intensity of illumination 2000Lx, 12 hour/day of light application time.
5. the taipei fritillary bulb bulb regenerating system method according to claim 1 established using seed as explant, feature
Be: the matrix in step S11 is specially coco bran: vegetable garden soil: perlite: vermiculite=1:3:2:1.
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| CN201910837236.3A CN110463606A (en) | 2019-09-05 | 2019-09-05 | A kind of taipei fritillary bulb bulb regenerating system method established using seed as explant |
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| CN201910837236.3A CN110463606A (en) | 2019-09-05 | 2019-09-05 | A kind of taipei fritillary bulb bulb regenerating system method established using seed as explant |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115104533A (en) * | 2022-07-20 | 2022-09-27 | 宜宾学院 | Callus induction culture medium and method with fritillaria stem as explant |
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| CN105638458A (en) * | 2015-12-29 | 2016-06-08 | 云南澈川生物科技有限公司 | Tissue cultivation method of Fritillaria cirrhosa D.Don |
| CN108668897A (en) * | 2018-05-18 | 2018-10-19 | 兰州市农业科技研究推广中心 | A kind of micro- numerous method of Fritillaria yuzhongensis |
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2019
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| US5801119A (en) * | 1996-12-31 | 1998-09-01 | Abbott Laboratories | Process for inhibiting stem elongation in bulbous plants and cut flowers therefrom |
| CN103283590A (en) * | 2012-03-01 | 2013-09-11 | 湖北省农科院中药材研究所 | Production method of fritillaria hupehensis seed ball |
| CN105638458A (en) * | 2015-12-29 | 2016-06-08 | 云南澈川生物科技有限公司 | Tissue cultivation method of Fritillaria cirrhosa D.Don |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN115104533A (en) * | 2022-07-20 | 2022-09-27 | 宜宾学院 | Callus induction culture medium and method with fritillaria stem as explant |
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