CN108719278A - A method of improving early pears stem apex drop vitrification method cryopreservation effect - Google Patents
A method of improving early pears stem apex drop vitrification method cryopreservation effect Download PDFInfo
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- CN108719278A CN108719278A CN201810545504.XA CN201810545504A CN108719278A CN 108719278 A CN108719278 A CN 108719278A CN 201810545504 A CN201810545504 A CN 201810545504A CN 108719278 A CN108719278 A CN 108719278A
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- stem apex
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N3/00—Preservation of plants or parts thereof, e.g. inhibiting evaporation, improvement of the appearance of leaves or protection against physical influences such as UV radiation using chemical compositions; Grafting wax
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- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Agronomy & Crop Science (AREA)
- Plant Pathology (AREA)
- Toxicology (AREA)
- Engineering & Computer Science (AREA)
- Dentistry (AREA)
- Wood Science & Technology (AREA)
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- Environmental Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The method for improving early pears stem apex drop vitrification method cryopreservation effect：Early pears stem apex is handled successively with six kinds of culture mediums containing carbon nanotube and graphene quantum dot, is remarkably improved survival rate after the jelly of stem apex.Compared with the prior art, early pears stem apex drop vitrification method cryopreservation effect is good, survival rate is high, stem apex easily regenerates by the present invention.
Description
Technical field：
The present invention relates to a kind of methods improving plant stem apex drop vitrification method cryopreservation effect, are exactly
A method of improving early pears stem apex drop vitrification method cryopreservation effect.
Background technology：
The excellent local jargonel germ plasm resource of the Jiangxi Zao Lishi, Shangrao Shangrao County reception room or parlour and the towns Tian Dundeng.Shangrao morning pears
There are two main breed is general, i.e. " serissa serissoide " and " Calusena lansium disappears ", wherein " serissa serissoide " quality is even better, be Shangrao morning pears most
Improved seeds, it is ripe early, and epidermis it is thin it is thin, meat is tender and crisp, core lacks that juice is more, taste is fragrant and sweet, the Qing Dynasty is once recommended as tribute.Ultralow temperature
Preservation is the product that Plantlet in vitro is combined with cryobiology, refers to that -80 DEG C of extremely low temperatures below preserve germ plasm resource
A whole set of biology techniques.Cryopreservation is a kind of safe and effective Germ-plasma resources protection approach, particularly with vegetative propagation
And the preservation of Recalcitrant Seeds plant, it is the approach of unique long-term preservation.Drop vitrification method has been compatible with traditional droplet vitrification and glass
The advantages of glass method, earliest droplet vitrification are the stem apex ultralow temperature for being put forward for the first time by Kartha etc. (1982), and being applied to cassava
It preserves, stem apex is placed in vitrifying drop, is gradually cooled down in sequencing cooling instrument.Drop vitrification is basic herein
The main program for going up and combining traditional glass method, i.e., handled material with loading liquid and vetrifying solution, then by material
It is placed on the aluminium-foil paper containing vitrifying drop and puts into a kind of ultra-low temperature preservation method of liquid nitrogen cryopreservation.This method has succeeded at present
It applies in the various plants such as Chinese yam, apple and potato.But that there is also survival rates is low for drop vitrification method cryopreservation, protects
Deposit the problems such as ineffective.For these problems, the present invention develops a kind of early pears stem apex drop vitrification method ultralow temperature of raising
The method of preservation effect can provide technical foundation for the long-term preservation of Shangrao morning pears germ plasm resource.
Invention content：
The object of the present invention is to provide a kind of survival rates, and the early pears stem apex drop vitrification method of the easily regenerated raising of high, stem apex is super
The method of Cord blood effect.Realizing the technical solution of the object of the invention is, a kind of early pears stem apex drop vitrification method of raising is super
The method of Cord blood effect, it is characterised in that have following steps：(1) in superclean bench, cut early pears stem apex and by its
It is inoculated in fluid nutrient medium one：MS+0.3-0.5g/L carbon nanotube+0.3-0.5g/L graphene quantum dot+0.3M sucrose carries out
Conventional preculture；(2) stem apex is transferred to fluid nutrient medium two：MS+2M glycerine+0.4M sucrose+0.3-0.5g/L carbon nanotubes+
0.3-0.5g/L graphene quantum dots carry out conventional loading；(3) stem apex is transferred to fluid nutrient medium three：PVS2+0.3-
0.5g/L carbon nanotube+0.3-0.5g/L graphene quantum dots carry out conventional dehydration；(4) stem apex is transferred to aluminum foil strip upper liquid
Body culture medium four：PVS2+0.3-0.5g/L carbon nanotube+0.3-0.5g/L graphene quantum dots carry out conventional drop vitrification
Method Liquid nitrogen storage；(5) after Liquid nitrogen storage, aluminum foil strip is taken out, fluid nutrient medium five is transferred to after routinely thawing：MS+
0.3-0.5g/L carbon nanotube+0.3-0.5g/L graphene quantum dot+1.2M sucrose carries out conventional wash；(6) stem apex is shifted
To solid regenerated culture medium six：MS+1-2mg/L Thidiazuron+0.1-0.3mg/L NAA+0.3-0.5g/L carbon nanotubes+0.3-
Material is put into culturing room after inoculation and carries out normal condition training by 0.5g/L graphene quantum dot+30g/L sucrose+7.5g/L agar
It supports, is remarkably improved survival rate after the jelly of stem apex.
Specific implementation mode：
In conjunction with following embodiments, the invention will be further described：
(1) it handles for the first time
In superclean bench, cuts early pears stem apex and be inoculated in fluid nutrient medium one：MS+0.3-0.5g/L carbon is received
Mitron+0.3-0.5g/L graphene quantum dot+0.3M sucrose carries out conventional preculture；
(2) it handles for second
Stem apex is transferred to fluid nutrient medium two：MS+2M glycerine+0.4M sucrose+0.3-0.5g/L carbon nanotubes+0.3-
0.5g/L graphene quantum dots carry out conventional loading；
(3) third time is handled
Stem apex is transferred to fluid nutrient medium three：PVS2+0.3-0.5g/L carbon nanotube+0.3-0.5g/L graphene quantums
Point carries out conventional dehydration；
(4) the 4th processing
Stem apex is transferred to fluid nutrient medium four on aluminum foil strip：PVS2+0.3-0.5g/L carbon nanotube+0.3-0.5g/L stones
Black alkene quantum dot carries out conventional drop vitrification method Liquid nitrogen storage；
(5) the 5th processing
After Liquid nitrogen storage, aluminum foil strip is taken out, fluid nutrient medium five is transferred to after routinely thawing：MS+0.3-0.5g/
L carbon nanotube+0.3-0.5g/L graphene quantum dot+1.2M sucrose carries out conventional wash；
(6) the 6th processing
Stem apex is transferred to solid regenerated culture medium six：MS+1-2mg/L Thidiazuron+0.1-0.3mg/L NAA+0.3-
0.5g/L carbon nanotube+0.3-0.5g/L graphene quantum dot+30g/L sucrose+7.5g/L agar, training is put into after inoculation by material
It supports room and carries out normal condition culture, be remarkably improved survival rate after the jelly of stem apex.
Claims (1)
1. the method for improving early pears stem apex drop vitrification method cryopreservation effect, it is characterised in that there is following steps：(1) exist
In superclean bench, cuts early pears stem apex and be inoculated in fluid nutrient medium one：MS+0.3-0.5g/L carbon nanotubes+0.3-
0.5g/L graphene quantum dot+0.3M sucrose carries out conventional preculture；(2) stem apex is transferred to fluid nutrient medium two：MS+2M
Glycerine+0.4M sucrose+0.3-0.5g/L carbon nanotube+0.3-0.5g/L graphene quantum dots carry out conventional loading；(3) by stem
Point is transferred to fluid nutrient medium three：PVS2+0.3-0.5g/L carbon nanotube+0.3-0.5g/L graphene quantum dots carry out conventional
Dehydration；(4) stem apex is transferred to fluid nutrient medium four on aluminum foil strip：PVS2+0.3-0.5g/L carbon nanotube+0.3-0.5g/L stones
Black alkene quantum dot carries out conventional drop vitrification method Liquid nitrogen storage；(5) after Liquid nitrogen storage, aluminum foil strip is taken out, by routinizing
Fluid nutrient medium five is transferred to after jelly：MS+0.3-0.5g/L carbon nanotube+0.3-0.5g/L graphene quantum dot+1.2M sucrose,
Carry out conventional wash；(6) stem apex is transferred to solid regenerated culture medium six：MS+1-2mg/L Thidiazuron+0.1-0.3mg/L NAA
+ 0.3-0.5g/L carbon nanotube+0.3-0.5g/L graphene quantum dot+30g/L sucrose+7.5g/L agar, by material after inoculation
It is put into culturing room and carries out normal condition culture, be remarkably improved survival rate after the jelly of stem apex.
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| CN201810545504.XA CN108719278A (en) | 2018-05-21 | 2018-05-21 | A method of improving early pears stem apex drop vitrification method cryopreservation effect |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104012524A (en) * | 2014-06-18 | 2014-09-03 | 中国农业科学院作物科学研究所 | Ultralow temperature storage and regeneration culture method of in vitro stem tip of Jerusalem artichoke |
| CN104170818A (en) * | 2014-09-15 | 2014-12-03 | 上海交通大学 | Method for optimizing vitrified cryopreservation effect of agapanthus embryonic calluses |
| CN104255707A (en) * | 2014-09-15 | 2015-01-07 | 上海交通大学 | Method for improving preservation effect of cymbidium type protocorm |
-
2018
- 2018-05-21 CN CN201810545504.XA patent/CN108719278A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104012524A (en) * | 2014-06-18 | 2014-09-03 | 中国农业科学院作物科学研究所 | Ultralow temperature storage and regeneration culture method of in vitro stem tip of Jerusalem artichoke |
| CN104170818A (en) * | 2014-09-15 | 2014-12-03 | 上海交通大学 | Method for optimizing vitrified cryopreservation effect of agapanthus embryonic calluses |
| CN104255707A (en) * | 2014-09-15 | 2015-01-07 | 上海交通大学 | Method for improving preservation effect of cymbidium type protocorm |
Non-Patent Citations (3)
| Title |
|---|
| SHU-MIN CHEN等: "CARBON NANOMATERIALS ENHANCE SURVIVAL OF Agapanthus praecox CALLUS AFTER CRYOPRESERVATION BY VITRIFICATION", 《CRYOLETTERS》 * |
| 吴昀等: "超低温保存植物种质资源的新途径-小滴玻璃化法", 《植物生理学报》 * |
| 王子成等: "带毒梨茎尖的玻璃化法超低温保存及其植株再生", 《河南大学学报》 * |
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Application publication date: 20181102 |