CN108812322A - A kind of processing method of emerald green arrowroot explant - Google Patents
A kind of processing method of emerald green arrowroot explant Download PDFInfo
- Publication number
- CN108812322A CN108812322A CN201810747337.7A CN201810747337A CN108812322A CN 108812322 A CN108812322 A CN 108812322A CN 201810747337 A CN201810747337 A CN 201810747337A CN 108812322 A CN108812322 A CN 108812322A
- Authority
- CN
- China
- Prior art keywords
- explant
- processing method
- arrowroot
- emerald green
- disinfection
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- 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
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The present invention relates to field of plant tissue culture, and in particular to a kind of processing method of emerald green arrowroot explant.The step of processing method, is as follows：1) explant selects：The base portion of stem of the seedling body with growing point or germination point of reservation emerald green arrowroot plant is as explant；2) explant pre-processes：Clean the explant surface smut；3) explant sterilizes：The use of volumetric concentration is successively 70%~75% ethanol solution and liquor natrii hypochloritis that volumetric concentration is 0.1~0.2% to the explant carry out infiltration disinfection to get.The sterile rate of sterilization method of the invention is high, and survival rate is high, operates relatively easy, it is easy to accomplish.
Description
Technical field
The present invention relates to field of plant tissue culture, and in particular to a kind of processing method of emerald green arrowroot explant.
Background technique
Emerald green arrowroot (Calathea ' Freddy) originates in Brazil, is Marantaceae, and Xiao's Maranta perennial evergreen draft is planted
Object.Being mainly characterized by blade surface has bottle-green speckle to be distributed, and plant height 15-20cm, blade oval, apex is tapering, blade
Celadon, along middle arteries, bottle-green speckle is evenly distributed on two sides, and the leaf margin that speckle continues up, blade back is green, and petiole is thin
It is long, green.Like warm and moist and half shade environment, be afraid of low temperature and dry-air blast, like subacidity soil, soil to drain, it is fertile, loose
Leaf mould or compost preferably, be the foliage plant for being very suitable to indoor placement.
Summary of the invention
The object of the present invention is to provide a kind of processing methods of emerald green arrowroot explant.
A kind of processing method of emerald green arrowroot explant, it is characterised in that steps are as follows：
1) explant selects：Retain the base portion of stem of the emerald green arrowroot plant with growing point or germination point as explant；
2) explant pre-processes：Clean the surface smut of the explant；
3) explant sterilizes：The use of volumetric concentration is successively 70%~75% ethanol solution and mass concentration be 0.1~
0.3% liquor natrii hypochloritis to the explant carry out infiltration disinfection to get.
Processing method of the present invention, the selection of step 1) explant, the emerald green arrowroot that preferred acquisition current year sprouts are strong
Health plant cuts off blade and root, retains the base portion for having growing point or the stem of sprouting bud point.
Preferably, the rudiment bar (with growing point or germination point) for choosing 1-2cm long is used as explant.Selection helps in this way
In the acquisition of the detoxification plant in follow-up cultivation.
Step 2) explant pretreatment of the present invention, cleans explant surface smut, it is preferred to use suitable concentration is washed
Clothing powder water impregnates 20-30min, then rinses in tap water down-flow water to cleaning, was advisable with 2-3 hours.Washing powder water concentration is excellent
It is selected as 3-5ppm.
Step 3) explant disinfection of the present invention, preferred operations are：The explant is peelled off after ethanol disinfection outer
Petiole is enclosed to tender shoots, is reused liquor natrii hypochloritis and is carried out disinfection.Excision is contacted with liquor natrii hypochloritis after hypochlorite disinfectant
Base portion tissue.
It preferably, the use of ethanol solution disinfecting time is preferably 20-40s,
And/or
Every liter of liquor natrii hypochloritis is preferably added to tween 3-4 drop, and the tween is Tween-20, uses sodium hypochlorite
Disinfecting time is 400-800s.
It is highly preferred that the volumetric concentration of the ethyl alcohol is 75%, disinfecting time 30s；The mass concentration of sodium hypochlorite is
0.2%, 3-4 drop polysorbas20, disinfecting time 420s is added in every liter of liquor natrii hypochloritis.
Aseptic water washing will be used after disinfection, thoroughly to remove sodium hypochlorite.It will be sufficiently stirred, protect when sterilizing and rinsing
Card material each section comes into full contact with disinfectant or sterile water.It is preferred that rinsed 5-7 times with sterile water, no less than 3min every time.
Sterilizing operation is carried out in desinfection chamber superclean bench.
Rinse water need to can carry out conventional choosing by disinfection treatment, those skilled in the art after water and disinfection in disinfectant
It selects, provides a kind of sterilization method herein, 121 DEG C, the disinfection with high pressure steam of 28min is handled.
The sterile rate of sterilization method of the invention is high, and survival rate is high, operates relatively easy, it is easy to accomplish.
The present invention provides a kind of tissue cultural method of emerald green arrowroot explant together, as follows：
Using the emerald green arrowroot explant by pretreatment and disinfection treatment, it is seeded in culture medium.At 20-25 DEG C
13h·d-1Illumination cultivation, preferably 23 DEG C.
Specific embodiment
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 bottle is chosen, is cleaned up, disinfection with high pressure steam is spare.
The preparation of disinfectant：
The sterile water of autoclave sterilization
0.1% liquor natrii hypochloritis (every liter is added 4 drop Tween-20s)
0.2% liquor natrii hypochloritis (every liter is added 4 drop Tween-20s)
70% ethyl alcohol
75% ethyl alcohol.
The preparation of instrument：
Sterilized filter paper, scalpel, scissors, tweezers.
Embodiment 1
The present embodiment provides a kind of processing method of emerald green arrowroot explant, concrete operations are：
1) explant selects：The healthy plant for acquiring the emerald green arrowroot of late November current year cuts off blade and root, retains emerald green
The seedling body base portion of leaf arrowroot plant and newborn tender shoots are as explant；
2) explant pre-processes：20min is impregnated with the washing powder water that concentration is 4ppm, flowing water is then carried out under tap water
2h is rinsed to clean to thorough；
3) explant sterilizes：Infiltration disinfection is carried out to the explant pre-processed in superclean bench, uses 75% second
Alcoholic solution sterilizes 30s, peels off peripheral petiole to tender shoots to explant, then with 0.2% sodium hypochlorite+Tween-20 solution to institute
It states explant to carry out disinfection 420s, be rinsed 7 times using sterile water, each 3min.
Embodiment 2
The present embodiment provides a kind of processing method of emerald green arrowroot explant, concrete operations are：
1) explant selects：The healthy plant for acquiring the emerald green arrowroot of late November cuts off blade and root, retains emerald green leaf bamboo
The seedling body base portion of taro plant and newborn tender shoots are as explant；
2) explant pre-processes：20min is impregnated with the washing powder water that concentration is 4ppm, flowing water is then carried out under tap water
2h is rinsed to clean to thorough；4ppm
3) explant sterilizes：Infiltration disinfection is carried out to the explant pre-processed in superclean bench, uses 70% second
Alcoholic solution sterilizes 30s, peels off peripheral petiole to tender shoots to explant, then with 0.2% sodium hypochlorite+polysorbas20 solution to described
Explant carries out disinfection 600s, is rinsed 7 times using sterile water, each 3min.
Embodiment 3
The present embodiment provides a kind of processing method of emerald green arrowroot explant, concrete operations are：
1) explant selects：The healthy plant for acquiring the emerald green arrowroot of late November cuts off blade and root, retains emerald green leaf bamboo
The seedling body base portion of taro plant and newborn tender shoots are as explant；
2) explant pre-processes：20min is impregnated with the washing powder water that concentration is 4ppm, flowing water is then carried out under tap water
2h is rinsed to clean to thorough；
3) explant sterilizes：Infiltration disinfection is carried out to the explant pre-processed in superclean bench, uses 75% second
Alcoholic solution sterilizes 30s, peels off peripheral petiole to tender shoots to explant, then with 0.1% sodium hypochlorite+polysorbas20 solution to described
Explant carries out disinfection 420s, is rinsed 7 times using sterile water, each 3min.
Embodiment 4
The present embodiment provides a kind of processing method of emerald green arrowroot explant, concrete operations are：
1) explant selects：The healthy plant for acquiring the emerald green arrowroot of late November cuts off blade and root, retains emerald green leaf bamboo
The seedling body base portion of taro plant and newborn tender shoots are as explant；
2) explant pre-processes：20min is impregnated with the washing powder water that concentration is 4ppm, flowing water is then carried out under tap water
2h is rinsed to clean to thorough；
3) explant sterilizes：Infiltration disinfection is carried out to the explant pre-processed in superclean bench, uses 75% second
Alcoholic solution sterilizes 20s, peels off peripheral petiole to tender shoots to explant, then with 0.2% sodium hypochlorite+polysorbas20 solution to described
Explant carries out disinfection 600s, is rinsed 7 times using sterile water, each 3min.
Embodiment 5
The present embodiment provides a kind of processing method of emerald green arrowroot explant, concrete operations are：
1) explant selects：The healthy plant for acquiring the emerald green arrowroot of late November cuts off blade and root, retains emerald green leaf bamboo
The seedling body base portion of taro plant and newborn tender shoots are as explant；
2) explant pre-processes：20min is impregnated with the washing powder water that concentration is 4ppm, flowing water is then carried out under tap water
2h is rinsed to clean to thorough；
3) explant sterilizes：Infiltration disinfection is carried out to the explant pre-processed in superclean bench, uses 75% second
Alcoholic solution sterilizes 40s, peels off peripheral petiole to tender shoots to explant, then with 0.2% sodium hypochlorite+polysorbas20 solution to described
Explant carries out disinfection 500s, is rinsed 7 times using sterile water, each 3min.
Comparative example 1
This comparative example provides a kind of processing method of emerald green arrowroot explant, and concrete operations are：
1) explant selects：The healthy plant for acquiring the emerald green arrowroot of late November cuts off blade and root, retains emerald green leaf bamboo
The seedling body base portion of taro plant and newborn tender shoots are as explant；
2) explant pre-processes：20min is impregnated with the washing powder water that concentration is 4ppm, flowing water is then carried out under tap water
2h is rinsed to clean to thorough；
3) explant sterilizes：Infiltration disinfection is carried out to the explant pre-processed in superclean bench, uses 75% second
Alcoholic solution sterilizes 30s, peels off peripheral petiole to tender shoots to explant, then with 0.3% sodium hypochlorite+polysorbas20 solution to described
Explant carries out disinfection 600s, is rinsed 7 times using sterile water, each 3min.
Test example 1
The preparation-obtained explant of the above various embodiments is inoculated into culture medium, the 13hd at 25 DEG C-1Illumination training
It supports.It is each to repeat 50, to guarantee 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.
It is as a result as follows through detecting：
| Embodiment 1 | Embodiment 2 | Embodiment 3 | Embodiment 4 | Embodiment 5 | Comparative example 1 | |
| Sterile rate | 100% | 100% | 90% | 96% | 100% | 100% |
| Survival rate | 96% | 86% | 88% | 88% | 82% | 12% |
Specifically, the 1 equal thorough in sterilization of resulting explant 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.
There is explantation tissue's browning necrosis phenomena in embodiment 2,4,5, can not obtain after individual explants are sterilized de-
Malicious plant；
There is explant disinfection and is not thorough phenomenon in embodiment 3, and the regeneration plant that carries disease germs germination slowly, forms runt seedling.
Wherein the whole Brown necrosis situations of comparative example 1 are serious, cannot obtain detoxification plant.
Although above having used general explanation, specific embodiment and test, 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 emerald green arrowroot explant, it is characterised in that：Steps are as follows：
1) explant selects：The base portion of stem of the seedling body with growing point or germination point of reservation emerald green arrowroot plant is as explant
Body；
2) explant pre-processes：Clean the explant surface smut；
3) explant sterilizes：It the use of volumetric concentration is successively 70%~75% ethanol solution and volumetric concentration is 0.1~0.2%
Liquor natrii hypochloritis to the explant carry out infiltration disinfection to get.
2. processing method according to claim 1, it is characterised in that：Step 1) the emerald green arrowroot selection current year sprouts
Healthy plant.
3. processing method according to claim 1 or 2, it is characterised in that：Step 1), the rudiment bar for choosing 1-2cm long are made
For explant.
4. processing method according to claim 1-3, it is characterised in that：Step 3) is, right after ethanol disinfection
The explant peels off peripheral petiole to tender shoots, reuses liquor natrii hypochloritis and carries out disinfection, after hypochlorite disinfectant excision with
The base portion tissue of liquor natrii hypochloritis's contact.
5. processing method according to claim 1-4, it is characterised in that：It is molten using the ethyl alcohol in step 3)
The liquid disinfectant time is 20-40s.
6. processing method according to claim 1-5, it is characterised in that：In step 3), sodium hypochlorite is used
Disinfecting time is 400-800s.
7. processing method according to claim 1-6, it is characterised in that：Every liter of liquor natrii hypochloritis addition
3-4 drop Tween-20 or Tween-80.
8. processing method according to claim 1-7, it is characterised in that：
Step 3) is to sterilize 20-30s using the ethanol solution of volumetric concentration 75%, peel off peripheral petiole extremely to the explant
Tender shoots, then carried out disinfection 400-420s with 0.2% sodium hypochlorite to the explant, wherein every liter of liquor natrii hypochloritis adds
Enter 3-4 drop polysorbas20.
9. processing method according to claim 1-8, it is characterised in that：
Step 3) is the ethanol disinfection time 30s for the use of volumetric concentration being 75%；Peripheral petiole is peelled off to tender to the explant
Bud reuses the sodium hypochlorite that mass concentration is 0.2% and sterilizes 420s to the explant, wherein every liter of liquor natrii hypochloritis
3-4 drop polysorbas20 is added.
10. processing method according to claim 1, it is characterised in that：Include the following steps：
1) explant selects：The healthy plant for acquiring emerald green arrowroot cuts off blade and root, retains the seedling body base of emerald green arrowroot plant
Portion and newborn tender shoots are as explant；
2) explant pre-processes：Thoroughly clean；
3) explant sterilizes：Infiltration disinfection is carried out to the explant pre-processed, sterilizes 30s using 75% ethanol solution, it is right
Explant peels off peripheral petiole to tender shoots, then is carried out disinfection with 0.2% sodium hypochlorite+Tween-20 solution to the explant
420s, after being rinsed using sterile water to get.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810747337.7A CN108812322A (en) | 2018-07-09 | 2018-07-09 | A kind of processing method of emerald green arrowroot explant |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810747337.7A CN108812322A (en) | 2018-07-09 | 2018-07-09 | A kind of processing method of emerald green arrowroot explant |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108812322A true CN108812322A (en) | 2018-11-16 |
Family
ID=64136562
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810747337.7A Pending CN108812322A (en) | 2018-07-09 | 2018-07-09 | A kind of processing method of emerald green arrowroot explant |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108812322A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110476811A (en) * | 2019-08-20 | 2019-11-22 | 广州市名卉景观科技发展有限公司 | A method of improving Ctenanthe oppenheimiana tissue-cultured seedling quality |
| CN110476812A (en) * | 2019-08-20 | 2019-11-22 | 广州市名卉景观科技发展有限公司 | A method of for Calathea roseopicta fast breeding |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101390494A (en) * | 2007-09-21 | 2009-03-25 | 天津市农业生物技术研究中心 | New method for obtaining ornamental arrowroot tissue culture sterile system |
| CN101796923A (en) * | 2010-03-26 | 2010-08-11 | 浙江传化生物技术有限公司 | Tissue culture rapid breeding method of ctenanthe oppenheimiana |
-
2018
- 2018-07-09 CN CN201810747337.7A patent/CN108812322A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101390494A (en) * | 2007-09-21 | 2009-03-25 | 天津市农业生物技术研究中心 | New method for obtaining ornamental arrowroot tissue culture sterile system |
| CN101796923A (en) * | 2010-03-26 | 2010-08-11 | 浙江传化生物技术有限公司 | Tissue culture rapid breeding method of ctenanthe oppenheimiana |
Non-Patent Citations (6)
| Title |
|---|
| M.PODWYSZYŃSKA: "Micropropagation of Calathea ornata Koern.", 《BIOLOGIA PLANTARUM》 * |
| 侯占铭等: "美丽竹芋的组织培养", 《植物生理学通讯》 * |
| 夏时云等: "不同消毒处理对竹芋组培无菌外植体获得的影响", 《天津农业科学》 * |
| 孙晓萍: "《观赏植物良种繁育技术》", 31 August 2009, 浙江人民出版社 * |
| 王吉等: "银叶竹芋的组织培养和快速繁殖", 《植物生理学通讯》 * |
| 王四敏: "竹芋和三色竹芋的试管繁殖研究", 《四川师范大学学报（自然科学版）》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110476811A (en) * | 2019-08-20 | 2019-11-22 | 广州市名卉景观科技发展有限公司 | A method of improving Ctenanthe oppenheimiana tissue-cultured seedling quality |
| CN110476812A (en) * | 2019-08-20 | 2019-11-22 | 广州市名卉景观科技发展有限公司 | A method of for Calathea roseopicta fast breeding |
| CN110476811B (en) * | 2019-08-20 | 2022-06-07 | 广州市名卉景观科技发展有限公司 | Method for improving quality of double-line arrowroot tissue culture seedlings |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105145359B (en) | Tissue culture and rapid propagation method for asparagus filicinus | |
| CN104813939B (en) | Method for constructing lotus regeneration system | |
| CN104855285B (en) | A kind of organic culture one-step-seedling formation abductive approach of polyploid Herba Anoectochili roxburghii | |
| CN103283595B (en) | Primary culture method of stem tip tissue of strawberry | |
| CN103651124B (en) | A kind of abductive approach of plant regeneration of zingiber officinale | |
| CN108812322A (en) | A kind of processing method of emerald green arrowroot explant | |
| CN108617503B (en) | Production method of ultrasonic-mediated colchicine double japonica rice chromosome | |
| CN106035083A (en) | Tissue culture method for rapid detoxification of purple potatoes | |
| CN107568066A (en) | A kind of potato stem tip detoxification method | |
| CN112931224A (en) | Tissue culture method of morinda officinalis | |
| CN116998278A (en) | Method for sterilizing coastal pine seeds for tissue culture | |
| CN106069784A (en) | A kind of method for tissue culture of Flos Caryophylli Fructus Gardeniae petal | |
| CN106577285A (en) | Method for inducing garlic callus tetraploid and regenerating test tube bulbs | |
| CN107318656B (en) | A kind of repetition culture poison-removing method of Potato Shoot-tips | |
| CN109691390A (en) | Wild rice stem seed callus cultivation | |
| CN109618928A (en) | A method of reducing indoor pot Chinese rose tissue culture explant body pollution | |
| CN106797890B (en) | A kind of culture medium and its method reducing leather leaf winged euonymus tissue culture pollution rate | |
| CN114902962A (en) | Living body intermittent disinfection method for clonal propagation of Epimedium sagittatum root tuber | |
| CN110896842B (en) | Efficient cultivation method of citrus virus-free etiolation stock and application thereof | |
| CN106234220A (en) | A kind of method of Beijing 553 Sweetpotato Viruses Elimination nursery | |
| CN110178728B (en) | Tissue culture and rapid propagation method for salvia miltiorrhiza bunge | |
| CN107027622B (en) | A kind of Herba Andrographitis Polyploid Induction Methods | |
| CN108668903A (en) | A kind of processing method of rye winter explant | |
| CN108668904A (en) | A kind of processing method of drought tuber of pinellia explant | |
| CN106688886A (en) | Explant disinfecting and cultivating method in taro stem tip tissue culture and plants |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20181116 |
|
| RJ01 | Rejection of invention patent application after publication |