CN108157184A - A kind of Lesley's new pteris fern seedling tissue culture and rapid propagation method - Google Patents
A kind of Lesley's new pteris fern seedling tissue culture and rapid propagation method Download PDFInfo
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- CN108157184A CN108157184A CN201810142486.0A CN201810142486A CN108157184A CN 108157184 A CN108157184 A CN 108157184A CN 201810142486 A CN201810142486 A CN 201810142486A CN 108157184 A CN108157184 A CN 108157184A
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- seedling
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- lesley
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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
- A01H4/001—Culture apparatus for tissue culture
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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
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
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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
- A01H4/008—Methods for regeneration to complete plants
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- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
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- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses a kind of tissue culture and rapid propagation methods of reproduction speed Lesley's new pteris fern seedling that is fast, at low cost and can keeping Lesley's Bird's Nest blade tip straight uniform bifurcated.The present invention includes the following steps：（1）Lesley's Bird's Nest root-like stock is taken as explant and carries out aseptic process, short grained protocorm is differentiated in incision；（2）Above-mentioned protocorm is taken to be placed in MS culture mediums to be cultivated, each protocorm differentiation goes out 3~4 new protocorms, carries out the Multiplying culture of aseptic seedling；（3）Protocorm is placed in MS cultures
Description
Technical field
The present invention relates to a kind of plant cultivation method more particularly to a kind of Lesley's new pteris ferns（Asplenium Nidus
“Lesley”）The tissue culture and rapid propagation method of seedling.
Background technology
Lesley's new pteris fern（Lesley, Aspleniaceae nest Cyclosorus Asplenium Nidus, original producton location：The Asia southeast,
Australian east, Indonesia, India and eastern Africa）As a member in new pteris fern family, blade tip besom
Shape bifurcated leaf shape, it is even more very popular.
The method that the plant division that existing Lesley's new pteris fern cultivation is usually used is bred, a new pteris fern 1 year
3~4 can be bred, because its breeding coefficient is low, far from the needs for meeting market.Therefore, the propagation method of new pteris fern at present
Mostly using spore seeding method and tissue culture method, and the tissue culture method of current new pteris fern is raw material also with spore
Progress is fast numerous, and concrete technology is：Being cut from sporophyll by the sporangium of sterilization, it is inoculated into after broken taking-up spore
On culture medium, spore germination is very fast, and germination rate is high.Original foliage after sprouting is inoculated on the MS culture mediums for do not add hormone and can obtain
To larger growth coefficient.Either spore seeding method, can be fast still using the tissue culture method that spore is raw material progress
Speed obtains a large amount of new pteris fern seedlings, but is carried out for Lesley's new pteris fern, sporogenesis and using spore for raw material fast
Numerous seedling has 30% or so, and perfect can not to inherit blade tip besom shape beauty bifurcated equal, therefore is sieved in planting process
Choosing can just obtain neat uniform Lesley's Bird's Nest.Again because Lesley's only blade tip crotch can just be collected into maturation
Spore, relatively common new pteris fern, it is also more relatively difficult to collect a large amount of spores.
Invention content
The technical problems to be solved by the invention are to overcome the deficiencies of the prior art and provide that a kind of reproduction speed is fast, cost
The tissue culture and rapid propagation method of Lesley's new pteris fern seedling that is low and can keeping Lesley's Bird's Nest blade tip straight uniform bifurcated.
The technical solution adopted in the present invention is that this method includes the following steps：
（1）Induction and differentiation：Lesley's Bird's Nest root-like stock is taken as explant and carries out aseptic process, aseptic process of learning from else's experience
Root-like stock, which is cut into several bulks and is put into the MS culture mediums added with 1~3mg/L of BA0.2~1.0mg/L and IAA, to be cultivated, temperature
It it is 22~24 degree, 1800~2000lux of illumination, daily 12 hours of light application time, after cultivating 18 weeks, incision differentiates little particle
Protocorm；
（2）The Multiplying culture of aseptic seedling：Above-mentioned protocorm is taken to be placed in the KT of the BA containing 0.2~0.5mg/L, 0.1~0.5mg/L
In the MS culture mediums of 3% sugar, after cultivating six weeks, each protocorm can differentiate 3~4 new protocorms, culture environment more
It is required that it is：22~24 degree, illumination 2500lux, daily illumination 12 hours；
（3）The transplanting domestication of aseptic seedling：Protocorm is placed in the activated carbon containing 3% sugar and 3g/L and the MS without hormone is cultivated
After 4 weeks are cultivated in base, every protocorm starts to grow 2~4 0.5 centimetre of seedlings, then for seedling to be placed in containing 0.5~1mg/L
IBA and 3% sugar MS culture mediums in cultivate surrounding, every seedling grows up to 1~1.5 centimetre of seedling, and has two or three black
Small fibrous root carries out transplanting domestication, and it is 5 to clean aseptic seedling root culture medium and be transplanted into peat soil and perlite ratio：1
Medium Culture, it is 18~25 degree, 3000~7000lux of illumination to keep temperature, the non-textile mulch of first week after transplanting, and control is wet
Degree 85~95% keeps leaf surface water mist not do.
Further, the step（1）In, it is to the step of Lesley's new pteris fern root-like stock progress aseptic process：By institute
State the leaf sheath and hair scale of the clean outside of root-like stock stripping, first with 75% ethanol postincubation 10 seconds, then impregnated in 0.1% mercuric chloride solution
It takes out within 8~10 minutes, then with aseptic water washing 4~5 times.
Further, the step（3）In it is further comprising the steps of：It opens bottle cap and aseptic seedling is made gradually to fit within 3~4 days before cultivation
Answer external condition.
The beneficial effects of the invention are as follows：The present invention is using the IAA's of BA and 1~3mg/L added with 0.2~1.0mg/L
The culture medium of MS, the root-like stock that can effectively facilitate Lesley's new pteris fern differentiate protocorm；Contain in addition, the present invention uses
The BA of 0.2~0.5mg/L, the KT of 0.1~0.5mg/L and 3% sugar MS as proliferated culture medium, the vigorous increasing of protocorm energy
It grows；Last protocorm is placed in the MS culture mediums of the activated carbon containing 3% sugar and 3g/L of no hormone that cultivate six weeks can be quick
Grow seedling, seedling is placed in the MS culture mediums of the sugar of the IBA containing 0.5~1.0mg/L and 3% that culture can Rapid Rooting；This
Invention directly induces protocorm by using root-like stock, and directly fast breeding grows seedling, simplified culture condition to protocorm again
Or the techniques such as training method, production cost can be reduced, keeps the uniformity of seedling, therefore Lesley's new pteris fern of the present invention is fast
Breeding method reproduction speed is fast, at low cost, keeps shape unified, reduces the screening operation in later stage.
Specific embodiment
The invention discloses a kind of Lesley's new pteris ferns（Asplenium Nidus“Lesley”）The tissue-culturing rapid propagation side of seedling
Method.The method of the present invention is further described with specific implementation steps below.
（One）Induction and differentiation
（1）Lesley's Bird's Nest root-like stock is taken as explant；
（2）The root-like stock is shelled to the leaf sheath and hair scale of clean outside, first with 75% ethanol postincubation 10 seconds, then 0.1% liter
It impregnates 8~10 minutes and takes out in mercury solution, then with aseptic water washing 4~5 times；
（3）The root-like stock after mercuric chloride of learning from else's experience disinfection is cut into the fritter of about 0.3cm or so, is put into added with 0.2~1.0mg/L
BA and 1~3mg/L IAA MS culture mediums in cultivation temperature be 22~24 degree, 1800~2000lux of illumination, light application time
Daily 12 hours, after cultivating 18 weeks, it is seen that incision differentiates short grained protocorm.
（Two）The Multiplying culture of aseptic seedling
（1）The protocorm of one one is taken to be placed in the sugar of the BA containing 0.2~0.5mg/L, the KT of 0.1~0.5mg/L and 3%
In MS culture mediums, after cultivating six weeks, each protocorm can differentiate 3~4 new protocorms more；
（2）Cultivation temperature requirement：22~24 degree, illumination 2500lux, daily illumination 12 hours.
（Three）The rooting culture of aseptic seedling
（1）After the protocorm is placed in containing four weeks is cultivated in 3% sugar and the activated carbon of 3g/L and the MS culture mediums without hormone,
Every protocorm starts to grow 2~4 0.5 centimetre of seedlings, then seedling is placed in the sugar of the IBA containing 0.5~1mg/L and 3%
Surrounding is cultivated in MS culture mediums, every seedling grows up to 1~1.5 centimetre of seedling, and has the small fibrous root of two or three black, can be moved
Plant domestication；
（2）It opens bottle cap and aseptic seedling is made gradually to adapt to external condition within 3~4 days before cultivation；
（3）It is 5 to clean aseptic seedling root culture medium and be transplanted into peat soil and perlite ratio：1 Medium Culture；
（4）It is 18~25 degree to keep temperature, low light condition（3000~7000lux of illumination）, use non-woven fabrics within first week after transplanting
Covering controls humidity 85~95%, and leaf surface water mist is kept not do.
It is compared with existing Lesley's new pteris fern seedling tissue culture method, the present invention is using the BA added with 0.2~1.0mg/L
With the culture medium of the MS of the IAA of 1~3mg/L, the root-like stock that can effectively facilitate Lesley's new pteris fern differentiates protocorm；Separately
Outside, the present invention uses the sugared MS for containing the BA of 0.2~0.5mg/L, the KT of 0.1~0.5mg/L and 3% as Multiplying culture
Base, protocorm can vigorous proliferation；Last protocorm is placed in the MS culture mediums of the activated carbon containing 3% sugar and 3g/L of no hormone
Middle culture can quickly grow seedling for six weeks, and seedling is placed in the MS culture mediums of sugar of the IBA containing 0.5~1.0mg/L and 3%
Culture can Rapid Rooting；The present invention directly induces protocorm by using root-like stock, and directly fast breeding is grown protocorm again
Seedling, the techniques such as simplified culture condition or training method, can reduce production cost, keep the uniformity of seedling, therefore the present invention
Lesley's new pteris fern quick-breeding method reproduction speed it is fast, it is at low cost, keep shape unified, reduce the screening operation in later stage.
Present invention can apply to Lesley's new pteris ferns to cultivate field.
Claims (3)
1. a kind of Lesley's new pteris fern seedling tissue culture and rapid propagation method, which is characterized in that this method includes the following steps：
（1）Induction and differentiation：Lesley's Bird's Nest root-like stock is taken as explant and carries out aseptic process, aseptic process of learning from else's experience
Root-like stock, which is cut into the MS culture mediums of IAA that are several blocky and being put into BA and 1~3mg/L added with 0.2~1.0mg/L, to be trained
It supports, temperature is 22~24 degree, 1800~2000lux of illumination, daily 12 hours of light application time, after cultivating 18 weeks, incision differentiation
Go out short grained protocorm；
（2）The Multiplying culture of aseptic seedling：Above-mentioned protocorm is taken to be placed in the KT of the BA containing 0.2~0.5mg/L, 0.1~0.5mg/L
In the MS culture mediums of 3% sugar, after cultivating six weeks, each protocorm can differentiate 3~4 new protocorms, culture environment more
It is required that it is：22~24 degree, illumination 2500lux, daily illumination 12 hours；
（3）The transplanting domestication of aseptic seedling：Protocorm is placed in the activated carbon containing 3% sugar and 3g/L and the MS without hormone is cultivated
After 4 weeks are cultivated in base, every protocorm starts to grow 2~4 0.5 centimetre of seedlings, then for seedling to be placed in containing 0.5~1mg/L
IBA and 3% sugar MS culture mediums in cultivate surrounding, every seedling grows up to 1~1.5 centimetre of seedling, and has two or three black
Small fibrous root carries out transplanting domestication, and it is 5 to clean aseptic seedling root culture medium and be transplanted into peat soil and perlite ratio：1
Medium Culture, it is 18~25 degree, 3000~7000lux of illumination to keep temperature, the non-textile mulch of first week after transplanting, and control is wet
Degree 85~95% keeps leaf surface water mist not do.
A kind of 2. Lesley's new pteris fern seedling tissue culture and rapid propagation method according to claim 1, which is characterized in that the step
（1）In, it is to the step of Lesley's new pteris fern root-like stock progress aseptic process：By the root-like stock shell clean outside leaf sheath and
Hair scale, first with 75% ethanol postincubation 10 seconds, then impregnate 8~10 minutes and take out in 0.1% mercuric chloride solution, then use sterile water
It rinses 4~5 times.
3. a kind of Lesley's new pteris fern seedling tissue culture and rapid propagation method according to claim 1, which is characterized in that its feature exists
In the step（3）In it is further comprising the steps of：It opens bottle cap and aseptic seedling is made gradually to adapt to external condition within 3~4 days before cultivation.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201810142486.0A CN108157184A (en) | 2018-02-11 | 2018-02-11 | A kind of Lesley's new pteris fern seedling tissue culture and rapid propagation method |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201810142486.0A CN108157184A (en) | 2018-02-11 | 2018-02-11 | A kind of Lesley's new pteris fern seedling tissue culture and rapid propagation method |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103548692A (en) * | 2013-11-04 | 2014-02-05 | 珠海市园艺研究所 | Tissue culture rapid propagation method for asplenium antiquum osaka germchit |
| CN104663443A (en) * | 2015-02-25 | 2015-06-03 | 杨惠才 | Construction method of in-vitro regeneration system of adiantum reniforme var.sinense |
| CN105145365A (en) * | 2015-09-22 | 2015-12-16 | 云南省农业科学院花卉研究所 | Green globular body tissue culture propagation method for Alsophila costularis |
-
2018
- 2018-02-11 CN CN201810142486.0A patent/CN108157184A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103548692A (en) * | 2013-11-04 | 2014-02-05 | 珠海市园艺研究所 | Tissue culture rapid propagation method for asplenium antiquum osaka germchit |
| CN104663443A (en) * | 2015-02-25 | 2015-06-03 | 杨惠才 | Construction method of in-vitro regeneration system of adiantum reniforme var.sinense |
| CN105145365A (en) * | 2015-09-22 | 2015-12-16 | 云南省农业科学院花卉研究所 | Green globular body tissue culture propagation method for Alsophila costularis |
Non-Patent Citations (1)
| Title |
|---|
| 曾智等: "海金沙孢子体快速繁殖体系的建立 ", 《湖北民族学院学报(自然科学版)》 * |
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