CN105165620A - Eggplant flower bud surface disinfection technology - Google Patents
Eggplant flower bud surface disinfection technology Download PDFInfo
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- CN105165620A CN105165620A CN201510648967.5A CN201510648967A CN105165620A CN 105165620 A CN105165620 A CN 105165620A CN 201510648967 A CN201510648967 A CN 201510648967A CN 105165620 A CN105165620 A CN 105165620A
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Abstract
The invention discloses an eggplant flower bud surface disinfection technology comprising steps as follows: collecting flower buds, pretreating the flower buds, disinfecting surfaces of the flower buds and absorbing moisture remaining on the surfaces of the flower buds. Flower buds of plants growing in different plots and different seasons are picked and taken back into a room under water loss prevention and low-temperature conditions, after sepal lobes are removed, the flower buds are disinfected with 75% alcohol for 1-5 min, then disinfected with 6.5% sodium hypochlorite for 10-20 min and washed with sterile water for 4-5 times, and anthers are peeled for follow-up texts after water is absorbed. The eggplant flower buds are pretreated, the disinfection time of a disinfection agent is adjusted, the surfaces of the eggplant flower buds from various sources are effectively disinfected, the pollution rate during primary culture and subculture of the eggplant anthers is greatly decreased, the culture efficiency of the anthers and pollen is improved, the cultured anthers can obtain embryoids on multiple culture media, and by means of the technology, the anthers have the pollution rate lower than 20% and can even avoid pollution completely.
Description
Technical field
The invention belongs to field of tissue culture, before relating to eggplant anther culture, bud is effectively sterilized, to improve the technology of eggplant anther culture success rate, disinfecting of the eggplant bud plucked under being applicable to different cultivars, different growing stage and different growth conditions.
Background technology
Eggplant (SolariummelongenaL), call eggplant, short melon, hang vegetable seeds, fall Soviet Union, draft or suffruticose, fruit can supply cooked food, and root, stem, leaf, seed all can be used as medicine.Eggplant originates in Tropical Asian, and all there is cultivation each province of China.
The biotechnology of taking as the leading factor with tissue cultures and genetic engineering that developed recently gets up, for shortening the breeding cycle and seed selection new quality variety open a new way, by flower pesticide and pollen cultures can obtain isozygoty in a large number monoploid, double haploid, after breeding practice, greatly can shorten breeding process.The beginning of the seventies, Riana etc. successfully pass through eggplant anther culture through callus approach seedling, no matter be that eggplant flower pesticide or pollen (microspore) are cultivated at present, all have many successful reports, but the domestic parent utilizing flower pesticide or pollen cultures to obtain, the report selecting New Eggplant Varieties is but also few.Trace it to its cause, mainly to produce the frequency of regeneration plant not high for eggplant flower pesticide or pollen cultures, and doubling monoploids frequency in regeneration plant is not high.In fact, aseptic and induce callus great-hearted flower pesticide or embryoid is not difficult from eggplant, difficulty is the surface sterilization technology of eggplant flower pesticide (bud).Therefore, when conventional surface disinfectant forbiddings such as mercuric chloride, how utilizing environment amenable alcohol and clorox effectively to sterilize to eggplant bud surface, is the key technology of successfully carrying out eggplant flower pesticide or pollen cultures.
Summary of the invention
In order to better solve the pollution problem occurred in flower pesticide and pollen cultures process, the invention provides a kind of eggplant bud surface sterilization technology efficiently.
Technical scheme provided by the invention is:
A kind of eggplant bud surface sterilization technology, the step comprised is as follows:
(1) bud is gathered: to choose Pollen stage be the monokaryon later stage to the early stage eggplant bud of double-core, is contained in plastic packaging bag, plastic packaging bag put into the ice chest that ice cube is housed and takes back indoor in time;
(2) bud pretreatment: taken out from plastic packaging bag by bud, without water or other liquid wash, directly removes the calyx lobe of eggplant bud outer wrapping;
(3) alcohol disinfecting, hypochlorite disinfectant: the eggplant bud that arranged is loaded in aseptic bottle, disinfects in alcohol successively, hypochlorite disinfectant;
(4) aseptic water washing, wipe dry are for subsequent use: sop up with the moisture of autoclaved blotting paper by eggplant flower pesticide adsorption.
Wherein, the time disinfected in alcohol is 1-5 minute, is 10-20 minute with hypochlorite disinfectant's time.
Preferably, summer and autumn, high temperature season disinfecting time was alcohol disinfecting 3 minutes, clorox 12 minutes.
Preferably, the alcohol disinfecting in season that Winter-Spring temperature is lower can extend to 5 minutes, and clorox is the longest reaches 20 minutes.
Wherein, want fast, will immediately moisture remaining on bud be blotted only after flushed with aseptic water washing.
The present invention, by the pretreatment to eggplant bud, regulates the disinfecting time of sterilization reagent, realizes the effective sterilization to eggplant bud surface, various source.Application the present invention, greatly reduce pollution rate when eggplant flower pesticide first generation and squamous subculture, improve the efficiency of flower pesticide and pollen cultures, the flower pesticide of cultivation can obtain embryoid on multiple medium.Utilize this technology, flower pesticide pollution rate, lower than 20%, even can reach 100% and not pollute.
Accompanying drawing explanation
Fig. 1: the design sketch of eggplant bud surface sterilization technology of the present invention.
Embodiment
Below in conjunction with specific embodiments and the drawings, the invention will be further described.
1. gather bud: eggplant plant can grow in greenhouse, booth or open country, gather the bud on growth early stage, mid-term and later stage plant, to choose Pollen stage be the monokaryon later stage to the early stage eggplant bud (petal splits base portion 0-2mm apart from calyx) of double-core, be contained in plastic packaging bag (anti-dehydration), plastic packaging bag is put into the ice chest that ice cube is housed and takes back indoor in time, put into refrigerator refrigeration 24-48h or directly carry out pre-treatment;
2. bud pre-treatment: bud is taken out from plastic packaging bag, without water or other liquid wash, directly remove the calyx lobe of eggplant bud outer wrapping, concrete operations are that calyx lobe is peelled off by the lines split along sepal, but calyx tube can not be divested;
3. bud sterilization: the eggplant bud arranged is loaded in aseptic bottle, in superclean bench or Biohazard Safety Equipment, with the absolute ethyl alcohol process 1-5min of 75%, a Tween-80 is added after pouring out absolute ethyl alcohol, then the clorox process 10-20min of 6.5% is used, period does not stop to rock, and finally uses aseptic water washing 4-5 time;
4, the moisture blotting bud remained on surface is for subsequent use: after aseptic water washing terminates, sop up with the moisture of autoclaved blotting paper by eggplant flower pesticide adsorption immediately, then the direct flower pesticide that strips from bud is inoculated, and the flower pesticide having little time to inoculate can be placed on refrigerator and cooled and hide 1-2d.
5. the effect of bud surface sterilization: application the present invention, even if in summer, from the eggplant bud being in growth latter stage, trialeurodes vaporariorum plant occurred frequently and plucking, the pollution rate of its flower pesticide is also no more than 20%, pollution rate is greatly reduced than the contrast of conventional method, improve the anther cultural success rate of eggplant, the flower pesticide of cultivation can obtain embryoid on multiple medium, finally obtains complete regenerated plant.
The above; it is only preferred embodiment of the present invention; not any pro forma restriction is done to the present invention; any those skilled in the art; do not departing within the scope of technical solution of the present invention; according to technical spirit of the present invention, any simple amendment that above embodiment is done, equivalently replace and improve, within the protection domain all still belonging to technical solution of the present invention.
Claims (5)
1. an eggplant bud surface sterilization technology, is characterized in that: the step comprised is as follows:
(1) bud is gathered: to choose Pollen stage be the monokaryon later stage to the early stage eggplant bud of double-core, is contained in plastic packaging bag, plastic packaging bag put into the ice chest that ice cube is housed and takes back indoor in time;
(2) bud pretreatment: taken out from plastic packaging bag by bud, without water or other liquid wash, directly removes the calyx lobe of eggplant bud outer wrapping;
(3) alcohol disinfecting, hypochlorite disinfectant: the eggplant bud that arranged is loaded in aseptic bottle, disinfects in alcohol successively, hypochlorite disinfectant;
(4) aseptic water washing, wipe dry are for subsequent use: sop up with the moisture of autoclaved blotting paper by eggplant flower pesticide adsorption.
2. eggplant bud surface sterilization technology as claimed in claim 1, it is characterized in that: the time disinfected in alcohol is 1-5 minute, is 10-20 minute with hypochlorite disinfectant's time.
3. eggplant bud surface sterilization technology as claimed in claim 1, is characterized in that: summer and autumn, high temperature season disinfecting time was alcohol disinfecting 3 minutes, clorox 12 minutes.
4. eggplant bud surface sterilization technology as claimed in claim 1, is characterized in that: the alcohol disinfecting in season that Winter-Spring temperature is lower can extend to 5 minutes, and clorox is the longest reaches 20 minutes.
5. eggplant bud surface sterilization technology as claimed in claim 1, is characterized in that: want fast, will be blotted only by moisture remaining on bud immediately after flushed with aseptic water washing.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201510648967.5A CN105165620A (en) | 2015-10-09 | 2015-10-09 | Eggplant flower bud surface disinfection technology |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510648967.5A CN105165620A (en) | 2015-10-09 | 2015-10-09 | Eggplant flower bud surface disinfection technology |
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| CN105165620A true CN105165620A (en) | 2015-12-23 |
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| CN201510648967.5A Pending CN105165620A (en) | 2015-10-09 | 2015-10-09 | Eggplant flower bud surface disinfection technology |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116806694A (en) * | 2023-08-16 | 2023-09-29 | 金陵科技学院 | Eggplant doubled haploid anther culture method |
| CN116849128A (en) * | 2023-08-23 | 2023-10-10 | 金陵科技学院 | Method for culturing anther of Kazakii autotetraploid plant |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101371650A (en) * | 2007-08-24 | 2009-02-25 | 中国农业科学院蔬菜花卉研究所 | Method for inducing dissociate microspore callus of eggplant and regenerating plant strain |
| CN103444552A (en) * | 2013-09-28 | 2013-12-18 | 武汉市蔬菜科学研究所 | Method for inducing eggplant anther to regenerate haplobiont |
| CN104542274A (en) * | 2014-07-03 | 2015-04-29 | 郑州市蔬菜研究所 | Culture medium of induced haplobiont for culturing eggplant anther and method of culture medium |
-
2015
- 2015-10-09 CN CN201510648967.5A patent/CN105165620A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101371650A (en) * | 2007-08-24 | 2009-02-25 | 中国农业科学院蔬菜花卉研究所 | Method for inducing dissociate microspore callus of eggplant and regenerating plant strain |
| CN103444552A (en) * | 2013-09-28 | 2013-12-18 | 武汉市蔬菜科学研究所 | Method for inducing eggplant anther to regenerate haplobiont |
| CN104542274A (en) * | 2014-07-03 | 2015-04-29 | 郑州市蔬菜研究所 | Culture medium of induced haplobiont for culturing eggplant anther and method of culture medium |
Non-Patent Citations (2)
| Title |
|---|
| 张玉苗: "不同茄子材料小孢子脱分化及影响因子研究", 《中国优秀硕士学位论文全文数据库 农业科技辑》 * |
| 潘羽丰: "茄子（Solanum melongena L.)花药培养诱导单倍体植株的研究", 《中国优秀硕士学位论文全文数据库 农业科技辑》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116806694A (en) * | 2023-08-16 | 2023-09-29 | 金陵科技学院 | Eggplant doubled haploid anther culture method |
| CN116849128A (en) * | 2023-08-23 | 2023-10-10 | 金陵科技学院 | Method for culturing anther of Kazakii autotetraploid plant |
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Application publication date: 20151223 |
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