CN108668904A - A kind of processing method of drought tuber of pinellia explant - Google Patents
A kind of processing method of drought tuber of pinellia explant Download PDFInfo
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- CN108668904A CN108668904A CN201810747357.4A CN201810747357A CN108668904A CN 108668904 A CN108668904 A CN 108668904A CN 201810747357 A CN201810747357 A CN 201810747357A CN 108668904 A CN108668904 A CN 108668904A
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- explant
- liquor natrii
- natrii hypochloritis
- ethanol solution
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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
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Abstract
The present invention relates to plant tissue cultivation fields, and in particular to a kind of processing method of drought tuber of pinellia explant.The processing method, its step are as follows：1) selection of explant：Using non-irrigated tuber of pinellia stem tuber as explant；2) explant pre-processes：The explant is cleaned；3) disinfection of explant：It is carried out disinfection successively to the explant using ethanol solution and liquor natrii hypochloritis.The processing method of explant provided by the invention is made that improvement compared with the existing technology, can turn out ideal explant, has good adaptability, survival rate high in tissue cultures.
Description
Technical field
The present invention relates to plant tissue cultivation fields, and in particular to a kind of drought tuber of pinellia the explant processing method.
Background technology
The non-irrigated tuber of pinellia [Pinellia ternate (Thunb.) Breit] alias corbie eye, water is beautiful, literary, three steps jump, fiber crops jade
Fruit etc., Araeceae Pinellia Tenore, herbaceos perennial.Complete stool is 15-30 centimetres high, and underground stem tuber is spherical in shape, is the master being used as medicine
Want part.Wild, except Inner Mongol, Xinjiang, Qinghai, Tibet region, all parts of the country are distributed.2300 meters of height above sea level is born in go down the hill
Ground, the careless slope on hills, barren beach, cheuch, small stream side, common dark and damp environment weedy again.Stem tuber is used as medicine, toxic, can be eliminating dampness and eliminating phlegm,
Stopping nausea and vomiting by lowering the adverse flow of QI, the raw furuncle that disappears；Cure mainly coughing with a lot of sputum, nausea and vomiting；The dense property middle ear of acute mastitis, acute and chronicization are controlled in external application
It is scorching.Animal doctor is controlling synanche Huang.
The tuber of pinellia seeks seed and bulbil or stem tuber breeding, therefore in Natural Population, existing seedling, and have by bulbil or fritter
New individual made of stem development.Then it is typical " of four generations under one roof " from physiological age, including by seed development
The seedling come, the plant developed by the first generation, the second generation or even third generation bulbil, and directly grown by stem tuber and
At plant.
Invention content
The object of the present invention is to provide a kind of processing method of non-irrigated tuber of pinellia explant and tissue cultural methods.
A kind of processing method of drought tuber of pinellia explant, its step are as follows：
1) selection of explant：Using non-irrigated tuber of pinellia stem tuber as explant；
2) explant pre-processes：The explant is cleaned；
3) disinfection of explant：It is carried out disinfection successively to the explant using ethanol solution and liquor natrii hypochloritis.
Preferred steps 3 of the present invention) concrete operations of disinfection of explant are：Use ethanol solution and sodium hypochlorite molten successively
Liquid sterilizes the explant, and is repeated once.
The sterile rate of repeated disinfection, the non-irrigated tuber of pinellia can be promoted largely, and because of separate operations, and there is no lasting extend to sterilize
Time, so being damaged to plant tissue smaller.
The selection of step 1) explant of the present invention, it is preferred to use the stem tuber of the non-irrigated tuber of pinellia of rest period.Using rest period
Non-irrigated tuber of pinellia upgrowth situation it is good, suitable for obtain detoxification regeneration plant.
Non-irrigated tuber of pinellia stem tuber, which is cut to 0.5cm × 0.5cm, to be advisable.
Step 2) explant pretreatment of the present invention, preferably uses washing powder water to impregnate, then washed down in tap water down-flow water.
Washing powder water concentration is 10-15ppm, and soaking time 10-20min is advisable, optimal with 15min；When tap water down-flow water rinses
Between 60-120min, it is more suitable with 90min.
The disinfection of step 3) explant of the present invention, it is preferred that the volumetric concentration of the ethanol solution be 70%~
75%, and/or, the mass concentration of the liquor natrii hypochloritis is 0.1~0.3%, and every liter of liquor natrii hypochloritis is added 3-4 drops and spits
Temperature.
Disinfecting time is preferably：Ethanol solution sterilizes 20~40s, and/or, liquor natrii hypochloritis sterilizes 650~800s.
Further preferably：
Step 3) includes：
3.1) it is carried out disinfection to the explant using ethanol solution and liquor natrii hypochloritis；
3.2) ethanol solution and liquor natrii hypochloritis is used to carry out disinfection the explant that step 3.1) sterilized.
It is 70%~75% that the ethanol solution in step 3.1) and step 3.2), which is each independently selected from volumetric concentration,
Ethanol solution；The liquor natrii hypochloritis is independently selected from the liquor natrii hypochloritis that mass concentration is 0.1~0.3%,
3-4 drop polysorbas20s are added in every liter of liquor natrii hypochloritis.
It is highly preferred that in step 3.1), the ethanol solution concentration is 70%；Disinfecting time 25-35s；The hypochlorous acid
A concentration of the 0.1% of sodium solution, disinfecting time 720-750s.
In step 3.2), the ethanol solution concentration is 75%；Disinfecting time 25-35s；The liquor natrii hypochloritis's is dense
Degree is 0.2%, disinfecting time 700-720s.
Aseptic water washing is used after disinfection every time, thoroughly to remove sodium hypochlorite.5~7 times are rinsed using sterile water, every time
No less than 3min can reach preferable effect.
Disinfectant and sterile water can be pre-processed according to the conventional selection of those skilled in the art, and the present invention provides herein
A kind of processing mode uses 121 DEG C, the disinfection with high pressure steam of 26min is handled.
The processing method of explant provided by the invention is made that improvement compared with the existing technology, can turn out ideal
Explant, have good adaptability in tissue cultures, survival rate is high.
Specific implementation mode
The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention..
Sterilizing utensil prepares：
250ml plastics tissue culture bottles are chosen, are cleaned up, disinfection with high pressure steam is spare.
The preparation of disinfectant：
The sterile water of autoclave sterilization,
70% ethanol solution,
75% ethanol solution,
0.1% liquor natrii hypochloritis (every liter is added 3 drop Tween-20s),
0.2% liquor natrii hypochloritis (every liter is added 3 drop Tween-20s),
0.3% liquor natrii hypochloritis (every liter is added 3 drop Tween-20s)
The preparation of instrument：
Sterilized filter paper, scalpel, tweezers.
Embodiment 1
The present embodiment provides a kind of processing methods of non-irrigated tuber of pinellia explant, and its step are as follows：
1) selection of explant：The non-irrigated tuber of pinellia stem tuber of rest period is acquired as explant, is cut to 0.5cm × 0.5cm；
2) explant pre-processes：15min is impregnated with a concentration of 10ppm washing powder water, then is rinsed in tap water down-flow water
90min；
3) disinfection of explant：30s is infiltrated using 70% ethanol solution, 0.1% liquor natrii hypochloritis infiltrates 720s, uses
Sterile water rinses 5 times, each 3min.
4) secondary sterilization：After removal surface tissue, 30s, 0.1% liquor natrii hypochloritis leaching are infiltrated using 75% ethanol solution
Moisten 700s, is rinsed 7 times, each 3min using sterile water, then cut off wound and thimerosal contact portion.
Embodiment 2
The present embodiment provides a kind of processing methods of non-irrigated tuber of pinellia explant, and its step are as follows：
With differing only in for embodiment 1：
3) disinfection of explant：30s is infiltrated using 70% ethanol solution, 0.2% liquor natrii hypochloritis infiltrates 720s, uses
Sterile water rinses 5 times, each 3min.
It can get detoxic seedling a large amount of and that tissue is lossless.
Embodiment 3
The present embodiment provides a kind of processing methods of non-irrigated tuber of pinellia explant, and its step are as follows：
With differing only in for embodiment 1：
3) disinfection of explant：30s is infiltrated using 75% ethanol solution, 0.2% liquor natrii hypochloritis infiltrates 720s, uses
Sterile water rinses 5 times, each 3min.
Embodiment 4
The present embodiment provides a kind of processing methods of non-irrigated tuber of pinellia explant, and its step are as follows：
With differing only in for embodiment 1：
3) disinfection of explant：30s is infiltrated using 75% ethanol solution, 0.3% liquor natrii hypochloritis infiltrates 600s, uses
Sterile water rinses 5 times, each 3min.
Embodiment 5
The present embodiment provides a kind of processing methods of non-irrigated tuber of pinellia explant, with differing only in for embodiment 1：
3) disinfection of explant：20s is infiltrated using 70% ethanol solution, 0.2% liquor natrii hypochloritis infiltrates 600s, uses
Sterile water rinses 5 times, each 3min.
Embodiment 6
The present embodiment provides a kind of processing methods of non-irrigated tuber of pinellia explant, with differing only in for embodiment 1：
3) disinfection of explant：20s is infiltrated using 75% ethanol solution, 0.3% liquor natrii hypochloritis infiltrates 600s, uses
Sterile water rinses 5 times, each 3min.
Embodiment 7
The present embodiment provides a kind of processing methods of non-irrigated tuber of pinellia explant, with differing only in for embodiment 1：
4) secondary sterilization：After removal surface tissue, 30s, 0.2% liquor natrii hypochloritis leaching are infiltrated using 70% ethanol solution
Moisten 720s, is rinsed 7 times, each 3min using sterile water, then cut off wound and thimerosal contact portion.
Embodiment 8
The present embodiment provides a kind of processing methods of non-irrigated tuber of pinellia explant, with differing only in for embodiment 1：
4) secondary sterilization：After removal surface tissue, 30s, 0.2% liquor natrii hypochloritis leaching are infiltrated using 70% ethanol solution
Moisten 720s, is rinsed 7 times, each 3min using sterile water, then cut off wound and thimerosal contact portion.
Embodiment 9
The present embodiment provides a kind of processing methods of non-irrigated tuber of pinellia explant, with differing only in for embodiment 1：
4) secondary sterilization：After removal surface tissue, 30s, 0.3% liquor natrii hypochloritis leaching are infiltrated using 75% ethanol solution
Moisten 600s, is rinsed 7 times, each 3min using sterile water, then cut off wound and thimerosal contact portion.
Embodiment 10
The present embodiment provides a kind of processing methods of non-irrigated tuber of pinellia explant, with differing only in for embodiment 1：
4) secondary sterilization：After removal surface tissue, 20s, 0.2% liquor natrii hypochloritis leaching are infiltrated using 70% ethanol solution
Moisten 600s, is rinsed 7 times, each 3min using sterile water, then cut off wound and thimerosal contact portion.
Embodiment 11
The present embodiment provides a kind of processing methods of non-irrigated tuber of pinellia explant, with differing only in for embodiment 1：
4) secondary sterilization：After removal surface tissue, 20s, 0.3% liquor natrii hypochloritis leaching are infiltrated using 75% ethanol solution
Moisten 600s, is rinsed 7 times, each 3min using sterile water, then cut off wound and thimerosal contact portion.
Embodiment 12
The present embodiment provides a kind of processing methods of non-irrigated tuber of pinellia explant, with differing only in for embodiment 1：
4) secondary sterilization：After removal surface tissue, 40s, 0.2% liquor natrii hypochloritis leaching are infiltrated using 70% ethanol solution
Moisten 720s, is rinsed 7 times, each 3min using sterile water, then cut off wound and thimerosal contact portion.
Test example 1
The preparation-obtained explant of the above various embodiments is inoculated into culture medium, the 13hd at 25 DEG C-1Illumination is trained
It supports.It is each to repeat 100, to ensure the repeatability of experiment.
After culture 10 days, observation culture situation.
Culture medium is MS+2.0mg/L 6-BA+0.1mg/L IAA+3.0g/L sucrose+0.7g/L agar, pH=5.8.
After testing, as a result as follows：
Specifically, the equal thorough in sterilization of explant of 1 gained of embodiment, is easily obtained the regeneration plant of health, growth
Point or bud point are damaged relatively light or do not have, and are conducive to the sprouting of bud and the extension growth of growing point.
Embodiment 2,3,4,6 occurs explantation tissue's browning necrosis phenomena, can not obtain detoxification plant；Embodiment 5,7,
10, which occur explant disinfection, is not thorough phenomenon, and the regeneration plant that carries disease germs germination is slow, forms runt seedling.
Although above having used general explanation, specific implementation mode and experiment, the present invention is made to retouch in detail
It states, but on the basis of the present invention, it can be made some modifications or improvements, this is apparent to those skilled in the art
's.Therefore, these modifications or improvements without departing from theon the basis of the spirit of the present invention, belong to claimed
Range.
Claims (10)
1. a kind of processing method of drought tuber of pinellia explant, its step are as follows：
1) selection of explant：Using non-irrigated tuber of pinellia stem tuber as explant；
2) explant pre-processes：The explant is cleaned；
3) disinfection of explant：It is carried out disinfection successively to the explant using ethanol solution and liquor natrii hypochloritis.
2. processing method according to claim 1, it is characterised in that：Step 3) includes：
3.1) it is carried out disinfection to the explant using ethanol solution and liquor natrii hypochloritis；
3.2) ethanol solution and liquor natrii hypochloritis is used to carry out disinfection the explant that step 3.1) sterilized.
3. processing method according to claim 1 or 2, it is characterised in that：The drought tuber of pinellia is the non-irrigated tuber of pinellia of rest period
Stem tuber.
4. according to claim 1-3 any one of them processing methods, it is characterised in that：It is described drought the tuber of pinellia be cut to 0.5cm ×
0.5cm。
5. according to claim 2-4 any one of them processing methods, it is characterised in that：Institute in step 3.1) and step 3.2)
It states ethanol solution and is each independently selected from the ethanol solution that volumetric concentration is 70%~75%.
6. according to claim 2-5 any one of them processing methods, it is characterised in that：Described in step 3.1) and step 3.2)
Liquor natrii hypochloritis is independently selected from the liquor natrii hypochloritis that mass concentration is 0.1~0.3%, every liter of liquor natrii hypochloritis
3-4 drop polysorbas20s are added.
7. according to claim 1-6 any one of them processing methods, it is characterised in that：Using the ethanol solution disinfection 20~
40s, and or, sterilize 650~800s using the liquor natrii hypochloritis.
8. according to claim 2-7 any one of them processing methods, it is characterised in that：In step 3.1), the ethanol solution
A concentration of 70%；Disinfecting time 25-35s；A concentration of the 0.1% of the liquor natrii hypochloritis, disinfecting time 720-750s.
9. according to claim 2-8 any one of them processing methods, it is characterised in that：In step 3.2), the ethanol solution
A concentration of 75%；Disinfecting time 25-35s；A concentration of the 0.2% of the liquor natrii hypochloritis, disinfecting time 700-720s.
10. processing method according to claim 1, it is characterised in that：Steps are as follows：
1) selection of explant：The non-irrigated tuber of pinellia stem tuber of rest period is acquired as explant, is cut to 0.5cm × 0.5cm；
2) explant pre-processes：The explant is cleaned to clean；
3) disinfection of explant：25-35s is infiltrated using 70% ethanol solution, 0.2% liquor natrii hypochloritis infiltrates 720-750s；
4) secondary sterilization：After removal surface tissue, 25-35s, 0.2% liquor natrii hypochloritis leaching are infiltrated using 75% ethanol solution
Moisten 700-720s, then cuts off wound and thimerosal contact portion；
3-4 drop polysorbas20s are added in every liter of liquor natrii hypochloritis.
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| CN201810747357.4A CN108668904A (en) | 2018-07-09 | 2018-07-09 | A kind of processing method of drought tuber of pinellia explant |
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| CN201810747357.4A CN108668904A (en) | 2018-07-09 | 2018-07-09 | A kind of processing method of drought tuber of pinellia explant |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113575334A (en) * | 2021-08-26 | 2021-11-02 | 南昌师范学院 | Pinellia ternata rapid propagation method based on organ direct generation approach |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1475107A (en) * | 2002-08-14 | 2004-02-18 | 北京金长河科技发展有限公司 | Normalization tissue culturing and fast breeding method of pinellia ternata |
| CN1799342A (en) * | 2005-12-30 | 2006-07-12 | 浙江省农业科学院 | Pinellia detoxification, tissue culture and quick propagation method |
| CN101766116A (en) * | 2010-02-04 | 2010-07-07 | 王康才 | Method for hybridizing pinellia tuber and propagating F1 |
-
2018
- 2018-07-09 CN CN201810747357.4A patent/CN108668904A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1475107A (en) * | 2002-08-14 | 2004-02-18 | 北京金长河科技发展有限公司 | Normalization tissue culturing and fast breeding method of pinellia ternata |
| CN1799342A (en) * | 2005-12-30 | 2006-07-12 | 浙江省农业科学院 | Pinellia detoxification, tissue culture and quick propagation method |
| CN101766116A (en) * | 2010-02-04 | 2010-07-07 | 王康才 | Method for hybridizing pinellia tuber and propagating F1 |
Non-Patent Citations (5)
| Title |
|---|
| JUNLI WANG等: "Effect of different plant growth regulators on micro-tuber induction and plant regeneration of Pinellia ternate (Thunb) Briet", 《PHYSIOLOGY AND MOLECULAR BIOLOGY OF PLANTS》 * |
| 宋艳梅等: "半夏组织培养快速繁殖研究", 《山东中医药大学学报》 * |
| 李秀霞: "《植物组织培养》", 30 June 2014, 东北大学出版社 * |
| 王艺霖等: "旱半夏组织培养技术的优化研究", 《江西农业学报》 * |
| 靳松等: "半夏组培快繁体系的建立", 《时珍国医国药》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113575334A (en) * | 2021-08-26 | 2021-11-02 | 南昌师范学院 | Pinellia ternata rapid propagation method based on organ direct generation approach |
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Application publication date: 20181019 |