CN101451119A - Phytolacca americana seed quick-speed germination method and its application in aseptic seedlings culture - Google Patents
Phytolacca americana seed quick-speed germination method and its application in aseptic seedlings culture Download PDFInfo
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- CN101451119A CN101451119A CNA2008102432339A CN200810243233A CN101451119A CN 101451119 A CN101451119 A CN 101451119A CN A2008102432339 A CNA2008102432339 A CN A2008102432339A CN 200810243233 A CN200810243233 A CN 200810243233A CN 101451119 A CN101451119 A CN 101451119A
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- phytolacca acinosa
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Abstract
The invention relates to the field of plant tissue culture, in particular to a method for quickly germinating phytolacca americana seeds and application of the method to aseptic seedling culture. The method comprises: soaking the phytolacca americana seeds into a concentrated sulfuric acid for a certain period, damaging the structure of phytolacca americana seed coats, breaking the sleep state of the seeds, and making the seeds be quickly subjected to imbibition and enter the germination state; simultaneously, utilizing the sterilization function of the concentrated sulfuric acid to perform sterilization treatment on the phytolacca americana seeds; subsequently, inoculating the aseptic phytolacca americana seeds on a culture medium selected by an experiment, adjusting the conditions such as temperature and illumination, and making the phytolacca americana seeds germinate quickly; and finally obtaining aseptic phytolacca americana seedlings. The method can quickly break through the sleep state of the phytolacca americana seeds, and greatly shorten the germination time and the seedling time of the phytolacca americana seeds. The invention provides a method for culturing the aseptic phytolacca americana seedlings for laboratory investigation.
Description
Technical field
The present invention relates to the plant tissue culture field, specifically relate to a kind of method and the application of this method in aseptic seedling is cultivated of Phytolacca acinosa seed quick-speed germination.
Technical background
Phytolacca acinosa is the per nnial herb with many effects, and people mainly are to pluck wild resource to the utilization of Phytolacca acinosa at present, and this mainly is because Phytolacca acinosa breeding difficulty is unfavorable for implant mass.Phytolacca acinosa generally adopts seminal propagation, but reasons such as, compact structure big owing to kind of skin hardness, resting stage are long cause the sprouting of Phytolacca acinosa seed often to reach for one to several months, and germination rate are not high, this makes the growth cycle of Phytolacca acinosa very long, several months and even several years could obtain the meta-bolites that people want.Sprout difficult problem at Phytolacca acinosa, some attempt to break Phytolacca acinosa Seed Dormancy state by methods such as photo-irradiation treatment, alternating temperature processing, potassium nitrate solution immersion and Gibberellins solution immersions, impel its fast-germination.But mostly have little effect.Experimental result is not the fast-germination that can't promote the Phytolacca acinosa seed, is exactly that germination rate is low excessively, can not large-scale application.Therefore, the problem of solution Phytolacca acinosa seed germination difficulty becomes the key issue of implant mass, application and research Phytolacca acinosa.
Summary of the invention
Sprout a difficulty and the long difficult problem of growth cycle in order to overcome Phytolacca acinosa, the invention provides a kind of Phytolacca acinosa seed quick-speed germination and aseptic seedling cultured method, solved problem effectively.
The principle of institute of the present invention foundation is: Phytolacca acinosa is the seminal propagation plant, such Phytolacca acinosa seed that its breeding reason difficult, that resting stage is long is not many people's guesses contains the material that suppresses seed germination, but because the really up to the mark and compact structure of Phytolacca acinosa kind skin, the imbibition that is unfavorable for seed causes the Phytolacca acinosa seed germination slow.At Phytolacca acinosa kind skin problem really up to the mark, the present invention adopts the vitriol oil to soak the Phytolacca acinosa seed regular hour, make its structure of the destroying Phytolacca acinosa kind skin inclusion of seed (but can not damage simultaneously), break the seed dormancy state, seed just can enter germinating by rapid imbibition like this.We utilize the germicidal action of the vitriol oil that Phytolacca acinosa is carried out disinfection simultaneously, obtain aseptic seed, obtain aseptic seedling through sterile propagation.
The technical solution adopted in the present invention is:
A kind of method of Phytolacca acinosa seed quick-speed germination is before the Phytolacca acinosa seed germination is cultivated, and soaks seed 5～30min, the wash and remove residual vitriol oil then with the vitriol oil.
The present invention also provides the application of above-mentioned quick germination method in the Phytolacca acinosa aseptic seedling is cultivated.Said application is with the seed with vitriol oil immersion treatment, behind the sterile distilled water wash and remove residual vitriol oil, seed is inserted substratum, carries out sterile culture, obtains the Phytolacca acinosa aseptic seedling.
The preferred MS substratum of above-mentioned said substratum+20g/L sucrose+7g/L agar.
Concrete operation steps is:
1, the short of Phytolacca acinosa seed sprouts and surface sterilization
Get the Phytolacca acinosa seed of mature and plump, flushing is 3 hours in flowing water, drains away the water, and the Phytolacca acinosa seed is soaked in 5～30min in the vitriol oil, topples over then concentrated sulfuric acid solution, washes seed 6～8 times with sterile distilled water, removes the residual vitriol oil.To place on the aseptic filter paper through the seed of soaking flushing to drain away the water, standby.
2, the cultivation of Phytolacca acinosa test-tube plantlet
To be inoculated in through the Phytolacca acinosa seed of above-mentioned processing in the 150mL culturing bottle of the 50mLMS solid medium of packing into, 4～5 every bottle, handle 6～7 bottles of inoculations for every kind; In 26 ± 1 ℃ of dark down cultivations, seed begins to sprout after three days, and all backs seed is finished germination process, culturing bottle is changed over to 26 ± 1 ℃ then, cultivates in the illumination box of intensity of illumination 1500～2000LUX, can obtain the Phytolacca acinosa aseptic seedling after about two weeks.
Adopt the inventive method can break Phytolacca acinosa Seed Dormancy state fast, shortened the sprouting of Phytolacca acinosa seed greatly and become the seedling time.And provide a Phytolacca acinosa aseptic seedling cultured method to laboratory study.
Embodiment
Employed in the present invention term unless other explanation is arranged, generally has the implication of those of ordinary skills' common sense.
Below in conjunction with specific embodiment, and comparable data is described the present invention in further detail.Should be understood that these embodiment just in order to demonstrate the invention, but not limit the scope of the invention by any way.
In following embodiment, various processes of Xiang Ximiaoshuing and method are not ordinary methods as known in the art.The source of agents useful for same, trade(brand)name and be necessary to list its moiety person indicate when occurring first that all used thereafter identical reagent if no special instructions, and is all identical with the content of indicating first.
Embodiment 1
Pluck the Phytolacca acinosa fruit season in Phytolacca acinosa fruit and seed maturity by the end of September, choose fully matured and do not have fruit airing under the sun of insect pest, treat that the Phytolacca acinosa fruit is shrivelled fully after, rub with hand and to make seed from fruit, come off out.Sort then, after the letter sorting seed is continued airing until its drying.Cold shone in the sack that good seed is loaded on good permeability be stored in shady and cool place.
Get the Phytolacca acinosa seed of mature and plump, flushing is 3 hours in flowing water, drains away the water.Therefrom get 200 seeds and pack in the reagent bottle of 100mL, pour the 50mL vitriol oil into, rolling is even gently, soaks 15min.Topple over then and the vitriol oil, water flushing seed 8 times is removed the residual vitriol oil.To place on the filter paper through the seed of soaking flushing to drain away the water, standby.
To sow seeds in the flowerpot that a certain amount of soil is housed through the Phytolacca acinosa seed of above-mentioned processing, room temperature remains on about 25 ℃, and the most of seed in a week back begins to sprout, Phytolacca acinosa bud and elongation rapidly, and after January, the Phytolacca acinosa seedling can reach the 15cm height.
Embodiment 2
Get the Phytolacca acinosa seed of mature and plump, flushing is 3 hours in flowing water, drains away the water.Therefrom get 100 seeds and pack in the reagent bottle of aseptic 100mL, pour the 30mL vitriol oil into, rolling is even gently, soaks 15min.Topple over then and the vitriol oil,, remove the residual vitriol oil with sterile distilled water flushing seed 8 times.To place on the aseptic filter paper through the seed of soaking flushing to drain away the water, standby.
Under aseptic condition, will be inoculated in the 150mL culturing bottle of the 50mLMS solid medium of packing into (MS substratum+2% sucrose+0.7% agar), 5 every bottle, inoculate 6 bottles through the Phytolacca acinosa seed of above-mentioned processing; Then they are placed 26 ± 1 ℃ of dark down cultivations, the germination rate of statistics seed changes culturing bottle over to 26 ± 1 ℃ then after the week, cultivates in the illumination box of intensity of illumination 2000LUX, can obtain the Phytolacca acinosa aseptic seedling after two weeks.
Data processing and statistical method
(1) germination rate (%)=(chitting piece number/seed sum) * 100%
(2) pollution rate (%)=(polluting number/inoculation sum) * 100%
(3) uncontaminated rate (%)=(uncontaminated number/inoculation sum) * 100%
(4) seedling rate (%)=(number of seedling/sprouting number) * 100%
Handle by data statistics, we obtain the result, adopt aforesaid method to handle and cultivation Phytolacca acinosa seed, and sprout time is 3 days the earliest, and the germination rate of the Phytolacca acinosa seed after the week can reach 96.7%, and pollution rate is 0.The Phytolacca acinosa aseptic seedling can reach the 10cm height after three weeks, and seedling rate is 100%.
Claims (4)
1, a kind of method of Phytolacca acinosa seed quick-speed germination is characterized in that, behind vitriol oil immersion Phytolacca acinosa seed 5～30min, the flushing seed is removed the residual vitriol oil, sprouts cultivation then.
2, the application of the method for the said Phytolacca acinosa seed quick-speed germination of claim 1 in the Phytolacca acinosa aseptic seedling is cultivated.
3, according to the said application of claim 2, it is characterized in that concrete grammar is: will behind the sterile distilled water wash and remove residual vitriol oil, seed be inserted substratum with the seed of vitriol oil immersion treatment, and carry out sterile culture, and obtain the Phytolacca acinosa aseptic seedling.
According to the said application of claim 2, it is characterized in that 4, wherein said substratum is: MS substratum+20g/L sucrose+7g/L agar.
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| Application Number | Priority Date | Filing Date | Title |
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| CNA2008102432339A CN101451119A (en) | 2008-12-26 | 2008-12-26 | Phytolacca americana seed quick-speed germination method and its application in aseptic seedlings culture |
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| CNA2008102432339A CN101451119A (en) | 2008-12-26 | 2008-12-26 | Phytolacca americana seed quick-speed germination method and its application in aseptic seedlings culture |
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| CN101451119A true CN101451119A (en) | 2009-06-10 |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101897294A (en) * | 2010-06-22 | 2010-12-01 | 福建农林大学 | Seed selection method for sweet potato seedling of two generators in one year |
| CN104025761A (en) * | 2014-06-04 | 2014-09-10 | 广西壮族自治区药用植物园 | Method for promoting germination of Phytolacca Americana seeds |
| CN104041394A (en) * | 2014-06-26 | 2014-09-17 | 中国科学院、水利部成都山地灾害与环境研究所 | Seedling cultivation and transplanting method for radix phytolaccae |
| CN104067724A (en) * | 2014-06-26 | 2014-10-01 | 中国科学院、水利部成都山地灾害与环境研究所 | Method for safely breaking pokeberry root seed episperm |
| CN104472062A (en) * | 2014-12-31 | 2015-04-01 | 包头医学院 | Method for promoting cymbariae seeds to germinate |
| CN107258154A (en) * | 2017-08-09 | 2017-10-20 | 青岛农业大学 | A kind of method that tissue culture is sterilized with soya seeds |
| CN107980634A (en) * | 2017-12-15 | 2018-05-04 | 贵州大学 | A kind of method for obtaining the suitable explant of lucid asparagus tissue-culturing rapid propagation |
| CN108668552A (en) * | 2018-04-16 | 2018-10-19 | 中山大学 | A kind of ion type rareearth tailings ground modifying agent and its restorative procedure |
-
2008
- 2008-12-26 CN CNA2008102432339A patent/CN101451119A/en active Pending
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101897294A (en) * | 2010-06-22 | 2010-12-01 | 福建农林大学 | Seed selection method for sweet potato seedling of two generators in one year |
| CN101897294B (en) * | 2010-06-22 | 2012-04-11 | 福建农林大学 | Seed selection method for sweet potato seedling of two generators in one year |
| CN104025761A (en) * | 2014-06-04 | 2014-09-10 | 广西壮族自治区药用植物园 | Method for promoting germination of Phytolacca Americana seeds |
| CN104041394A (en) * | 2014-06-26 | 2014-09-17 | 中国科学院、水利部成都山地灾害与环境研究所 | Seedling cultivation and transplanting method for radix phytolaccae |
| CN104067724A (en) * | 2014-06-26 | 2014-10-01 | 中国科学院、水利部成都山地灾害与环境研究所 | Method for safely breaking pokeberry root seed episperm |
| CN104041394B (en) * | 2014-06-26 | 2016-01-20 | 中国科学院、水利部成都山地灾害与环境研究所 | A kind of nursery of Phytolacca acinosa and trasplanting method |
| CN104472062A (en) * | 2014-12-31 | 2015-04-01 | 包头医学院 | Method for promoting cymbariae seeds to germinate |
| CN107258154A (en) * | 2017-08-09 | 2017-10-20 | 青岛农业大学 | A kind of method that tissue culture is sterilized with soya seeds |
| CN107980634A (en) * | 2017-12-15 | 2018-05-04 | 贵州大学 | A kind of method for obtaining the suitable explant of lucid asparagus tissue-culturing rapid propagation |
| CN108668552A (en) * | 2018-04-16 | 2018-10-19 | 中山大学 | A kind of ion type rareearth tailings ground modifying agent and its restorative procedure |
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