CN107046864A - A kind of method of Seed sterilization during orchid aseptic seeding - Google Patents
A kind of method of Seed sterilization during orchid aseptic seeding Download PDFInfo
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- CN107046864A CN107046864A CN201710018272.8A CN201710018272A CN107046864A CN 107046864 A CN107046864 A CN 107046864A CN 201710018272 A CN201710018272 A CN 201710018272A CN 107046864 A CN107046864 A CN 107046864A
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- seed
- sterilization
- pod
- fruit pod
- thimerosal
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
- A01C1/08—Immunising seed
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- Life Sciences & Earth Sciences (AREA)
- Soil Sciences (AREA)
- Environmental Sciences (AREA)
- Pretreatment Of Seeds And Plants (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The sterilizing methods of seed when sterilizing methods and Fruit pod when not ftractureed the invention discloses a kind of method of Seed sterilization during orchid aseptic seeding, including Fruit pod ftracture：When Fruit pod does not ftracture, pod is removed into persitent perianth, stamen post, carpopodium, after being cleaned through running water, it is placed in 5% liquor natrii hypochloritis and sterilizes 20 minutes, with 75% alcohol surface sterilization 3 minutes, calcination removes pod surface alcohol directly on alcolhol burner, completes sterilization；When Fruit pod ftractures, seed is positioned in 300 mesh nylon net bags of 3cm × 4cm sizes, heat sealing, nylon net bag is placed in 10.0ml centrifuge tubes, then thimerosal is added into centrifuge tube, thimerosal liquid level was not had nylon wire, 20 40min are placed, with several times of aseptic water washing, sterilization are completed.Disinfecting time can effectively be controlled, it is ensured that surface sterilization effect, and easy to operate, be a kind of suitable for Orchid Seeds and the sterilization method of similar minute seeds on seed vitality without influence.
Description
Technical field
The present invention relates to a kind of method of Seed sterilization during plant seeding, more particularly to during a kind of orchid aseptic seeding
The method of Seed sterilization.
Background technology
The fruit of orchid is capsule, is ftractureed after capsule is ripe under field conditions (factors), discharges seed.Because capsule is
Enclosed construction, its seed cleaning sterile when not ftractureing directly can carry out carry out surface sterilization, its internal kind to Fruit pod
Son can be directly used for axenic germination.But during the crossbreeding of orchid, due to carelessness, fail to harvest Fruit pod in time, often
The Fruit pod maturity period can be run into once the phenomenon to cracking, or orchid Fruit pod ftractures in transporting procedures, once cracking seed is easy
It is contaminated.
Because Orchid Seeds are very tiny and light weight, conventional surface sterilization method is complex, and Disinfection Effect
It is poor, the problems such as easily causing loss and the uneven disinfecting time of seed, and then influence its axenic germination success rate.For small
The experimental technique of the surface of the seed sterilization existing some researchs at present, but there is no the suitable sterilization method for Orchid Seeds
Research.Therefore suitable Orchid Seeds surface sterilization technology, has for orchid axenic germination and industrial seedling rearing
Significance.
The content of the invention
It is an object of the invention to provide a kind of method of Seed sterilization during orchid aseptic seeding.
The purpose of the present invention is achieved through the following technical solutions：
The method of Seed sterilization during the orchid aseptic seeding of the present invention, including sterilizing methods of Fruit pod when not ftractureing and
The sterilizing methods of seed when Fruit pod ftractures：
When Fruit pod does not ftracture, pod is removed into persitent perianth, stamen post, carpopodium, after being cleaned through running water, 5% chlorine is placed in
Sterilized 20 minutes in acid sodium solution, with 75% alcohol surface sterilization 3 minutes, calcination removes pod surface directly on alcolhol burner
Alcohol, completes sterilization；
When Fruit pod ftractures, seed is positioned in 300 mesh nylon net bags of 3cm × 4cm sizes, heat sealing, by nylon
Mesh bag is placed in 10.0ml centrifuge tubes, then adds thimerosal into centrifuge tube, thimerosal liquid level was not had nylon wire, places 20-
40min, with several times of aseptic water washing, completes sterilization.
As seen from the above technical solution provided by the invention, orchid aseptic seeding provided in an embodiment of the present invention
When Seed sterilization method, can effectively control disinfecting time, it is ensured that surface sterilization effect, and easy to operate, to seed vitality without
Influence, is a kind of sterilization method suitable for Orchid Seeds and similar minute seeds.
Embodiment
The technical scheme in the embodiment of the present invention is clearly and completely described below, it is clear that described embodiment
Only a part of embodiment of the invention, rather than whole embodiments.Based on embodiments of the invention, ordinary skill
The every other embodiment that personnel are obtained under the premise of creative work is not made, belongs to protection scope of the present invention.
The method of Seed sterilization during the orchid aseptic seeding of the present invention, its preferably embodiment is：
The sterilizing methods of seed when sterilizing methods and Fruit pod when not ftractureed including Fruit pod ftracture：
When Fruit pod does not ftracture, pod is removed into persitent perianth, stamen post, carpopodium, after being cleaned through running water, 5% chlorine is placed in
Sterilized 20 minutes in acid sodium solution, with 75% alcohol surface sterilization 3 minutes, calcination removes pod surface directly on alcolhol burner
Alcohol, completes sterilization；
When Fruit pod ftractures, seed is positioned in 300 mesh nylon net bags of 3cm × 4cm sizes, heat sealing, by nylon
Mesh bag is placed in 10.0ml centrifuge tubes, then adds thimerosal into centrifuge tube, thimerosal liquid level was not had nylon wire, places 20-
40min, with several times of aseptic water washing, completes sterilization.
The composition of the thimerosal is：The Tween-20 of sodium hypochlorite+0.01% of the weight/mass percentage compositions of 7-8ml 2%.
With the seed rinsed after the agar solution autoclavings of 1-2ml 0.1% after the sterilizing in nylon net bag, and and seed
Mix, obtain seed suspension liquid, then seed suspension liquid is drawn to media surface with liquid-transfering gun, culture medium is sealed, water
Square to rotating and culturing ware or blake bottle, seed is evenly distributed in media surface, cultivated.
The method of Seed sterilization during the orchid aseptic seeding of the present invention, including sterilizing methods of Fruit pod when not ftractureing and
The sterilizing methods of seed when Fruit pod ftractures.Its seed cleaning sterile when not ftractureing, directly can carry out surface to Fruit pod and disappear
Poison.Split dehiscent fruit pod needs to carry out sterilization treatment to seed, and Orchid Seeds are small, and quality is extremely light, in aseptic seeding
When, conventional surface sterilizing methods are complex, and sterilization effect is poor, easily cause the loss of seed.The present invention is to orchid kind
Son carries out disinfection, and can effectively control disinfecting time, it is ensured that surface sterilization effect, and easy to operate, on seed vitality without influence, is
It is a kind of to be applied to Orchid Seeds and the sterilization method of similar minute seeds.
Specific embodiment：
The sterilizing of seed when sterilizing and Fruit pod when not ftractureed including Fruit pod are ftractureed.
When Fruit pod does not ftracture, pod is removed into persitent perianth, stamen post, carpopodium, after being cleaned through running water, 5% chlorine is placed in
Sterilized 20 minutes in acid sodium solution, with 75% alcohol surface sterilization 3 minutes, calcination removes pod surface directly on alcolhol burner
Alcohol, can thus complete sterilization.
When Fruit pod ftractures, sterilization method includes thimerosal processing and sterile two steps of water washing, in superclean bench
It is upper to carry out.
1) seed is positioned in 300 mesh nylon net bags of 3cm × 4cm sizes, heat sealing.Nylon net bag is placed in
In 10.0ml centrifuge tubes, then into centrifuge tube add thimerosal (composition of thimerosal is：7-8ml 2% (weight/mass percentage composition)
The Tween-20 of sodium hypochlorite+0.01%), thimerosal liquid level was not had nylon wire, cover the lid of centrifuge tube, place 20-40min,
Period, which shakes, to be mixed 3-5 times；Finally the bottom up of centrifuge tube is tilted, thimerosal is flowed out.
2) wash：To step 1) centrifuge tube in add sterilized water, sterilized water flows into centrifugation bottom of the tube and submerges Buddhist nun immediately
Imperial bag, uses hand rotated several times；The bottom up of centrifuge tube is tilted again, sterilized water is flowed out.Such repeated washing 4-5 times.
To the seed of sterilization.
Rinsed with the agar solutions of 1-2ml 0.1% (0.1g agar is dissolved in 100ml water, and autoclaving) in nylon net bag
Seed, and the above-mentioned agar solutions of 1-2ml 0.1% and seed are mixed, obtain seed suspension liquid；Then it will be planted with liquid-transfering gun
Sub- suspension is drawn to media surface, and culture medium is sealed, horizontal direction rotating and culturing ware (or blake bottle), makes seed uniform
Ground is distributed in media surface (in order to later statistics), is cultivated.Thickness of sowing is with every bottle of seed amount with substantially
It is degree that culture medium, which can be touched, and the normal growth after germination percentage and germination can be influenceed when too close.
The foregoing is only a preferred embodiment of the present invention, but protection scope of the present invention be not limited thereto,
Any one skilled in the art is in the technical scope of present disclosure, the change or replacement that can be readily occurred in,
It should all be included within the scope of the present invention.Therefore, protection scope of the present invention should be with the protection model of claims
Enclose and be defined.
Claims (3)
1. a kind of method of Seed sterilization during orchid aseptic seeding, it is characterised in that sterilizing when not ftractureed including Fruit pod
The sterilizing methods of seed when method and Fruit pod cracking：
When Fruit pod does not ftracture, pod is removed into persitent perianth, stamen post, carpopodium, after being cleaned through running water, 5% sodium hypochlorite is placed in
Sterilized 20 minutes in solution, with 75% alcohol surface sterilization 3 minutes, calcination removes pod surface wine directly on alcolhol burner
Essence, completes sterilization；
When Fruit pod ftractures, seed is positioned in 300 mesh nylon net bags of 3cm × 4cm sizes, heat sealing, by nylon net bag
It is placed in 10.0ml centrifuge tubes, then thimerosal is added into centrifuge tube, thimerosal liquid level was not had nylon wire, places 20-
40min, with several times of aseptic water washing, completes sterilization.
2. the method for Seed sterilization during orchid aseptic seeding according to claim 1, it is characterised in that the sterilization
The composition of liquid is：The Tween-20 of sodium hypochlorite+0.01% of the weight/mass percentage compositions of 7-8ml 2%.
3. the method for Seed sterilization during orchid aseptic seeding according to claim 1 or 2, it is characterised in that use 1-
The seed rinsed after the agar solution autoclavings of 2ml 0.1% after the sterilizing in nylon net bag, and mixed with seed, obtain seed
Suspension, then seed suspension liquid drawn to media surface, culture medium is sealed with liquid-transfering gun, horizontal direction rotating and culturing
Ware or blake bottle, make seed be evenly distributed in media surface, are cultivated.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710018272.8A CN107046864A (en) | 2017-01-10 | 2017-01-10 | A kind of method of Seed sterilization during orchid aseptic seeding |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710018272.8A CN107046864A (en) | 2017-01-10 | 2017-01-10 | A kind of method of Seed sterilization during orchid aseptic seeding |
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| CN201710018272.8A Pending CN107046864A (en) | 2017-01-10 | 2017-01-10 | A kind of method of Seed sterilization during orchid aseptic seeding |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107926212A (en) * | 2017-12-13 | 2018-04-20 | 甘肃省农业科学院林果花卉研究所 | A kind of sterilization method of the tone fruit trees fruit and seed such as peach, Lee, apricot |
| CN109362523A (en) * | 2018-10-29 | 2019-02-22 | 张志安 | A kind of globe amaranth method for culturing seedlings |
| CN110402642A (en) * | 2019-08-03 | 2019-11-05 | 湖北金水源生物科技有限公司 | A kind of sterilization method of Cremastra appendiculata dehiscent capsule |
| CN116762653A (en) * | 2023-07-27 | 2023-09-19 | 江苏农牧科技职业学院 | Method for rapidly obtaining pinellia ternate seedlings through seeds |
Citations (2)
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| CN101352120A (en) * | 2008-08-15 | 2009-01-28 | 中国科学院植物研究所 | Method for sterilizing surface of plant seed |
| CN105325287A (en) * | 2015-11-24 | 2016-02-17 | 长江大学 | Disinfection and inoculation method of microphyte seeds and pollen |
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2017
- 2017-01-10 CN CN201710018272.8A patent/CN107046864A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101352120A (en) * | 2008-08-15 | 2009-01-28 | 中国科学院植物研究所 | Method for sterilizing surface of plant seed |
| CN105325287A (en) * | 2015-11-24 | 2016-02-17 | 长江大学 | Disinfection and inoculation method of microphyte seeds and pollen |
Non-Patent Citations (2)
| Title |
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| 曹孜义等: "《实用植物组织培养技术教程》", 31 December 1996, 甘肃科学技术出版社 * |
| 李志英等: "《主要热带作物种质资源离体快繁技术规程》", 31 December 2007, 中国农业出版社 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107926212A (en) * | 2017-12-13 | 2018-04-20 | 甘肃省农业科学院林果花卉研究所 | A kind of sterilization method of the tone fruit trees fruit and seed such as peach, Lee, apricot |
| CN109362523A (en) * | 2018-10-29 | 2019-02-22 | 张志安 | A kind of globe amaranth method for culturing seedlings |
| CN110402642A (en) * | 2019-08-03 | 2019-11-05 | 湖北金水源生物科技有限公司 | A kind of sterilization method of Cremastra appendiculata dehiscent capsule |
| CN116762653A (en) * | 2023-07-27 | 2023-09-19 | 江苏农牧科技职业学院 | Method for rapidly obtaining pinellia ternate seedlings through seeds |
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