CN106305428B - A kind of elaeagnus mollis bud multiplication and plant regeneration method - Google Patents
A kind of elaeagnus mollis bud multiplication and plant regeneration method Download PDFInfo
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- CN106305428B CN106305428B CN201610780875.7A CN201610780875A CN106305428B CN 106305428 B CN106305428 B CN 106305428B CN 201610780875 A CN201610780875 A CN 201610780875A CN 106305428 B CN106305428 B CN 106305428B
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- 241001228114 Elaeagnus mollis Species 0.000 title claims abstract description 36
- 241000196324 Embryophyta Species 0.000 title claims abstract description 16
- 238000011069 regeneration method Methods 0.000 title claims abstract description 14
- 229920001817 Agar Polymers 0.000 claims description 16
- 239000008272 agar Substances 0.000 claims description 16
- 229930006000 Sucrose Natural products 0.000 claims description 15
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 15
- 239000005720 sucrose Substances 0.000 claims description 14
- 239000001963 growth medium Substances 0.000 claims description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 7
- 235000013399 edible fruits Nutrition 0.000 claims description 6
- 239000007788 liquid Substances 0.000 claims description 6
- 230000006698 induction Effects 0.000 claims description 4
- 239000002609 medium Substances 0.000 claims description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 3
- 239000003599 detergent Substances 0.000 claims description 3
- 238000007598 dipping method Methods 0.000 claims description 3
- 230000010355 oscillation Effects 0.000 claims description 3
- 229920000136 polysorbate Polymers 0.000 claims description 3
- 238000005406 washing Methods 0.000 claims description 3
- 229960002523 mercuric chloride Drugs 0.000 claims description 2
- LWJROJCJINYWOX-UHFFFAOYSA-L mercury dichloride Chemical compound Cl[Hg]Cl LWJROJCJINYWOX-UHFFFAOYSA-L 0.000 claims description 2
- 240000000111 Saccharum officinarum Species 0.000 claims 2
- 235000007201 Saccharum officinarum Nutrition 0.000 claims 2
- 238000000034 method Methods 0.000 abstract description 12
- 206010020649 Hyperkeratosis Diseases 0.000 abstract description 5
- 230000001939 inductive effect Effects 0.000 abstract description 3
- 230000035755 proliferation Effects 0.000 abstract description 3
- 230000008929 regeneration Effects 0.000 abstract description 3
- 238000004500 asepsis Methods 0.000 abstract 1
- 229960004793 sucrose Drugs 0.000 description 8
- 239000012877 elongation medium Substances 0.000 description 5
- GVJHHUAWPYXKBD-UHFFFAOYSA-N (±)-α-Tocopherol Chemical compound OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 description 4
- 238000012545 processing Methods 0.000 description 4
- UUTKICFRNVKFRG-WDSKDSINSA-N (4R)-3-[oxo-[(2S)-5-oxo-2-pyrrolidinyl]methyl]-4-thiazolidinecarboxylic acid Chemical compound OC(=O)[C@@H]1CSCN1C(=O)[C@H]1NC(=O)CC1 UUTKICFRNVKFRG-WDSKDSINSA-N 0.000 description 3
- 229930192334 Auxin Natural products 0.000 description 3
- 239000002363 auxin Substances 0.000 description 3
- 239000005556 hormone Substances 0.000 description 3
- 229940088597 hormone Drugs 0.000 description 3
- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 235000019198 oils Nutrition 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- OYHQOLUKZRVURQ-HZJYTTRNSA-N Linoleic acid Chemical compound CCCCC\C=C/C\C=C/CCCCCCCC(O)=O OYHQOLUKZRVURQ-HZJYTTRNSA-N 0.000 description 2
- 229930003427 Vitamin E Natural products 0.000 description 2
- 238000009395 breeding Methods 0.000 description 2
- 230000001488 breeding effect Effects 0.000 description 2
- 239000003246 corticosteroid Substances 0.000 description 2
- 229960001334 corticosteroids Drugs 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- WIGCFUFOHFEKBI-UHFFFAOYSA-N gamma-tocopherol Natural products CC(C)CCCC(C)CCCC(C)CCCC1CCC2C(C)C(O)C(C)C(C)C2O1 WIGCFUFOHFEKBI-UHFFFAOYSA-N 0.000 description 2
- 230000035784 germination Effects 0.000 description 2
- 235000020778 linoleic acid Nutrition 0.000 description 2
- OYHQOLUKZRVURQ-IXWMQOLASA-N linoleic acid Natural products CCCCC\C=C/C\C=C\CCCCCCCC(O)=O OYHQOLUKZRVURQ-IXWMQOLASA-N 0.000 description 2
- 239000002689 soil Substances 0.000 description 2
- 235000015112 vegetable and seed oil Nutrition 0.000 description 2
- 235000019165 vitamin E Nutrition 0.000 description 2
- 229940046009 vitamin E Drugs 0.000 description 2
- 239000011709 vitamin E Substances 0.000 description 2
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 241001508399 Elaeagnus Species 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 208000031226 Hyperlipidaemia Diseases 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- GAMYVSCDDLXAQW-AOIWZFSPSA-N Thermopsosid Natural products O(C)c1c(O)ccc(C=2Oc3c(c(O)cc(O[C@H]4[C@H](O)[C@@H](O)[C@H](O)[C@H](CO)O4)c3)C(=O)C=2)c1 GAMYVSCDDLXAQW-AOIWZFSPSA-N 0.000 description 1
- 230000003712 anti-aging effect Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 208000011775 arteriosclerosis disease Diseases 0.000 description 1
- RCTYPNKXASFOBE-UHFFFAOYSA-M chloromercury Chemical compound [Hg]Cl RCTYPNKXASFOBE-UHFFFAOYSA-M 0.000 description 1
- 230000019771 cognition Effects 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 235000013681 dietary sucrose Nutrition 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 230000035558 fertility Effects 0.000 description 1
- 229930003944 flavone Natural products 0.000 description 1
- -1 flavone compound Chemical class 0.000 description 1
- 235000011949 flavones Nutrition 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
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- 239000007921 spray Substances 0.000 description 1
- 235000021122 unsaturated fatty acids Nutrition 0.000 description 1
- 150000004670 unsaturated fatty acids Chemical class 0.000 description 1
- VHBFFQKBGNRLFZ-UHFFFAOYSA-N vitamin p Natural products O1C2=CC=CC=C2C(=O)C=C1C1=CC=CC=C1 VHBFFQKBGNRLFZ-UHFFFAOYSA-N 0.000 description 1
- 239000002023 wood Substances 0.000 description 1
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
- A01H4/008—Methods for regeneration to complete plants
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- 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 methods of elaeagnus mollis bud multiplication and plant regeneration, this method is according to sprouting is easily sprouted at elaeagnus mollis axillary bud the characteristics of, seedling is obtained using ripe elaeagnus mollis seed asepsis sprouting, tender stem segments of the clip with axillary-bud or top-bud are explant, promote axillary bud sprouting, elongation by the method for tissue cultures, the healthy and strong shoot with certain degree of lignification is obtained, to realize the purpose of proliferation, then realizes plant regeneration through culture of rootage.The method that the present invention uses bud multiplication avoids in traditional elaeagnus mollis method for tissue culture and the problem of browning easily occurs during callus and adventitious bud inducing, easy to operate, feasible.
Description
Technical field
The invention belongs to Tree Organization culture technique fields, and in particular to a kind of elaeagnus mollis bud multiplication and plant regeneration
Method.
Background technology
Elaeagnus mollis (Elaeagnusmollis Diels) is the perennial deciduous tree of Elaeangnaceae Elaeagnus or filling
Wood is the distinctive ancient plant kind of China that quaternary glacier is survived.Elaeagnus mollis seed oil is full of nutrition, balanced, comprehensive, rich
Pufa-containing, wherein linoleic acid content are up to 45%, linoleic acid be a kind of humans and animals body itself cannot synthesize and
The unsaturated fatty acid that must be absorbed from food has the function of the illnesss such as preventing hypertension, hyperlipidemia, vascular sclerosis.Wing
The content of vitamin E is particularly surprising in fruit oil seeds benevolence, up to 1558.1mg/100g.The growth and development of vitamin E and human body is close
Cut phase is closed, and has significant anti-oxidant, anti-aging effects.It is rich in flavone compound in elaeagnus mollis leaf, can be cardiovascular diseases
Disease medicament development provides source.In addition, because its nectar amount is big, material is fine and closely woven solid, and well developed root system is simultaneously lived with nodule nitrogen fixation
Property, it is cultivated in nectariferous plant, Timber stands, water and soil conservation improves soil fertility aspect, before elaeagnus mollis also has certain application
Scape.
Past due to elaeagnus mollis lack correctly cognition and Substantial evaluation realize, the felling of the mankind, reclaim wasteland with
And the extensive unconfined natural community's production for directly picking the productions activities such as wild elaeagnus mollis seed oil expression to elaeagnus mollis
Severe jamming destruction is given birth to.Other than human factor, the biological characteristics such as natural habitat of elaeagnus mollis itself fragility is narrow；Knot
Real rate is low, easy depauperation；Under natural conditions, seed longeivity is short, and low wait of germination percentage also exacerbates the in imminent danger of its population.Therefore,
It is necessary to take measures to restore, expand Elaeagnus Mollis Population, the genetic resources to protect this important and excellent rare oil
Expect seeds, makes full use of its economic, ecology and scientific research value.
The breeding of elaeagnus mollis usually has seminal propagation and root division.But seed longeivity under elaeagnus mollis natural conditions
Short, germination percentage is low, and root division has that breeding coefficient is not high.Tissue cultures are one and are widely used to production in fact
The biotechnology trampled, but in elaeagnus mollis tissue culture procedures callus induction and when adventitious bud browning easily occurs, our rule is kept away
This problem is opened, the characteristics of using sprouting is easily sprouted at elaeagnus mollis axil, using the method for bud multiplication, axillary bud is promoted to sprout
Hair, elongation, realize plant regeneration.
Invention content
Goal of the invention：For the deficiencies in the prior art, the object of the present invention is to provide a kind of elaeagnus mollis normal buds
Proliferation and plant regeneration method are avoided easily going out during callus and adventitious bud inducing in traditional elaeagnus mollis method for tissue culture
The problem of existing browning, realizes elaeagnus mollis vegetative propagation.
Technical solution：In order to achieve the above-mentioned object of the invention, the technical solution adopted by the present invention is：
A kind of elaeagnus mollis bud multiplication and plant regeneration method, include the following steps：
(1) well-developed ripe elaeagnus mollis fruit is chosen, outer layer nut shells is peelled off, is cleaned, sterilized, is sterile
Processing；Then internal layer kind skin is removed under anatomical lens, is placed in 3/4MS+ sucrose 30g/L+ agar 7g/L culture mediums, and it is full to obtain axillary bud
Full, healthy and strong seedling；
(2) seedling of acquisition is cut into 2~4 axillary-bud or top-buds, the stem section of long 3~4cm, level is inoculated in 3/
4MS+TDZ 0.006~0.008mg/L+NAA 0.1mg/L+ sucrose 30g/L+ agar 7g/L culture mediums handle 7~10d；
(3) stem section that step (2) obtains is inoculated in 3/4MS+BA0.4~0.5mg/L+NAA0.05~0.07mg/ vertically
L+ sucrose 30g/L+ agar 7g/L medium cultures, subculture is primary when 20d；
(4) it waits for that normal bud is elongated to 6cm or more in step (3), after there is certain degree of lignification, is inoculated in root media
Continue to cultivate in 1/2MS+IBA 1.5mg/L+NAA 0.01~0.03+ sucrose 25g/L+ agar 7g/L, carries out rooting induction；Most
Seedling is obtained eventually.
In step (1), after rinsing 1h with a small amount of liquid detergent and 84 medicining liquid dipping 10min and flowing water, used in super-clean bench
70% alcohol impregnates 40s, then impregnates 20min, aseptic water washing 4~5 with appropriate 0.1% mercuric chloride and a small amount of Tween~20, oscillation
It is secondary.
In step (3), culture medium is 3/4MS+BA 0.5mg/L+NAA0.05~0.07mg/L.
In step (4), root media is 1/2MS+IBA 0.01~0.03mg/L+KT of 1.5mg/L+NAA 0.01mg/
L+ sucrose 25g/L+ agar 7g/L.
In step (4), root media is 1/2MS+IBA 1.5mg/L+NAA 0.03mg/L+KT 0.01mg/L+ sucrose
25g/L+ agar 7g/L.
Advantageous effect：Compared with prior art, the method for the invention using bud multiplication can avoid traditional samara oil
Easily there is the problem of browning during callus and adventitious bud inducing in tree method for tissue culture；Stem section with axillary bud or normal bud connects
Kind handles 7~10d on the bud Primary culture culture medium comprising 0.006~0.008mg/LTDZ and 0.1mg/LNAA, can be effective
Promote axillary bud sprouting at axil, shortens growing-seedling period；The BA0.5mg/L's and NAA0.05~0.07mg/L of bud elongation medium
Hormone combination, it is the spray with certain altitude, rugosity and degree of lignification that can effectively facilitate normal bud elongation, is conducive to take root.
Description of the drawings
Fig. 1 is bud Primary culture pictorial diagram；
Fig. 2 is bud elongation cultivation results figure；
Fig. 3 is figure of taking root.
Specific implementation mode
With reference to specific embodiment, the present invention is further illustrated.
The condition of culture that following embodiment uses is：Intensity of illumination (2000lx), light application time 14h/d, cultivation temperature 25
DEG C or so；Medium pH 5.7~5.8.
Embodiment 1
A kind of elaeagnus mollis bud multiplication and plant regeneration method, include the following steps：
(1) acquisition of aseptic seedling：Well-developed ripe elaeagnus mollis fruit is chosen, it is hard that physicomechanical processes peel off outer layer
Shell is impregnated in super-clean bench with 70% alcohol after rinsing 1h with a small amount of liquid detergent and 84 medicining liquid dipping 10min and flowing water
40s, then 20min, aseptic water washing 4~5 times are impregnated with appropriate 0.1% mercury chloride and a small amount of Tween~20, oscillation；Then exist
Fruit internal layer kind skin is removed under anatomical lens, is placed in 3/4MS culture mediums, additional saccharose 30g/L, agar 7g/L, acquisition axillary bud is full,
Healthy and strong seedling.
(2) selection of bud primary culture medium：By the seedling of acquisition, it is cut into 2~4 axillary-bud or top-buds of band and is about 3~4cm
Stem section, as shown in Figure 1, level is inoculated in 3/4MS minimal mediums, additional hormone concentration and media is 0.004,0.006,
0.008,0.01,0.02,0.05,0.1mg/LTDZ and 0.1mg/L NAA, handling duration 5d, 10d, 15d.
(3) selection of bud elongation medium：The stem section that 5d, 10d, 15d are handled in above-mentioned bud primary culture medium is inoculated in
3/4MS minimal mediums, additional hormone concentration and media be 0.1,0.3,0.5,0.7,0.9mg/L BA and 0.03,0.05,
0.07,0.09mg/L NAA, 20d subculture are primary.
During bud Primary culture, TDZ concentration is bigger, and processing time is longer, and stem section expands degree of deformity and vitrifying journey
Degree is more serious, and it is also corresponding further difficult to go to progress bud elongation culture in bud elongation medium.TDZ>0.01mg/L, when processing
Between>When 10d, stem section excessively expands deformity, and vitrifying is serious；When 0.004<TDZ<When 0.08mg/L, 5~10d of processing time, stem
Section occur it is a degree of expands, sprouting is sprouted at the axil without axillary bud originally, goes to additional 0.5mg/L BA, 0.05~
The bud elongation medium of 0.07mg/LNAA, the normal bud number that is averaged per stem section increase to 4~8, as shown in Figure 2.
Consider factors, the buds such as explant vitrifying degree, bud proliferation rate, bud elongation difficulty or ease, cultivation cycle length to open
Dynamic culture medium is advisable with a concentration of 0.006~0.008mg/L of TDZ, 7~10d of handling duration；Bud elongation medium is a concentration of with BA
A concentration of 0.05~0.07mg/L of 0.5mg/L, NAA is best, when 35d or so, sprout height about 4~6cm.
(4) culture of rootage：It extends to after with certain rugosity, height (about 6cm or more) and degree of lignification, is inoculated with after bud
In 1/2MS minimal mediums, IBA, NAA, KT and 25g/L sucrose are added, statistical data when 35d.Rooting rate=take root successfully
The seedling number of seedling number/inoculation；Item number=total number of root/of taking root is taken root successful seedling number.
Root media and Corticosteroids situation and corresponding result are as shown in table 1 and Fig. 3.The great shadow of auxin concentration
Ring the induced efficiency of the growing way and root of elaeagnus mollis tissue-cultured seedling.Relatively low (the IBA of auxin concentration<1.0mg/L, NAA<0.03mg/L)
When, it does not take root or takes root and is slow, radical amount is few；Higher (the IBA of auxin concentration>1.5mg/L, NAA>When 0.03mg/L), seedling base portion
Callus is easily formed, fallen leaves are serious, cause to take root unsuccessfully.
1 root media of table and Corticosteroids and corresponding experimental result
Consider explant rooting rate, the quality of root, the growing way of seedling, it is IBA1.5mg/L to be suitble to the culture medium taken root,
IAA0.03mg/L, is added or not additional 0.01mg/L KT, and rooting rate is up to 30.0%~36.7%.
Claims (5)
1. a kind of elaeagnus mollis bud multiplication and plant regeneration method, which is characterized in that include the following steps：
(1) well-developed ripe elaeagnus mollis fruit is chosen, outer layer nut shells is peelled off, is cleaned, sterilized, sterile place
Reason；Then internal layer kind skin is removed under anatomical lens, is placed in 3/4MS+ sucrose 30g/L+ agar 7g/L culture mediums, and it is full to obtain axillary bud
Full, healthy and strong seedling；
(2) seedling of acquisition is cut into 2~4 axillary-bud or top-buds, the stem section of long 3~4cm, level is inoculated in 3/4MS+
TDZ 0.006~0.008mg/L+NAA 0.1mg/L+ sucrose 30g/L+ agar 7g/L culture mediums handle 7~10d；
(3) stem section that step (2) obtains is inoculated in 3/4MS+BA0.4~0.5mg/L+NAA0.05~0.07mg/L+ sugarcanes vertically
Sugared 30g/L+ agar 7g/L medium cultures, subculture is primary when 20d；
(4) it waits for that normal bud is elongated to 6cm or more in step (3), after there is certain degree of lignification, is inoculated in root media 1/
Continue to cultivate in 2MS+IBA 1.5mg/L+NAA 0.01~0.03+ sucrose 25g/L+ agar 7g/L, carries out rooting induction；Finally
Obtain seedling.
2. elaeagnus mollis bud multiplication according to claim 1 and plant regeneration method, it is characterised in that：In step (3),
Culture medium is 3/4MS+BA 0.5mg/L+NAA0.05~0.07mg/L.
3. a kind of elaeagnus mollis bud multiplication and plant regeneration method, which is characterized in that include the following steps：
(1) well-developed ripe elaeagnus mollis fruit is chosen, outer layer nut shells is peelled off, is cleaned, sterilized, sterile place
Reason；Then internal layer kind skin is removed under anatomical lens, is placed in 3/4MS+ sucrose 30g/L+ agar 7g/L culture mediums, and it is full to obtain axillary bud
Full, healthy and strong seedling；
(2) seedling of acquisition is cut into 2~4 axillary-bud or top-buds, the stem section of long 3~4cm, level is inoculated in 3/4MS+
TDZ 0.006~0.008mg/L+NAA 0.1mg/L+ sucrose 30g/L+ agar 7g/L culture mediums handle 7~10d；
(3) stem section that step (2) obtains is inoculated in 3/4MS+BA0.4~0.5mg/L+NAA0.05~0.07mg/L+ sugarcanes vertically
Sugared 30g/L+ agar 7g/L medium cultures, subculture is primary when 20d；
(4) it waits for that normal bud is elongated to 6cm or more in step (3), after there is certain degree of lignification, is inoculated in root media 1/
Continue to cultivate in 2MS+IBA 1.5mg/L+NAA 0.01~0.03mg/L+KT 0.01mg/L+ sucrose 25g/L+ agar 7g/L,
Carry out rooting induction；It is final to obtain seedling.
4. elaeagnus mollis bud multiplication according to claim 3 and plant regeneration method, it is characterised in that：In step (4),
Root media is 1/2MS+IBA 1.5mg/L+NAA 0.03mg/L+KT 0.01mg/L+ sucrose 25g/L+ agar 7g/L.
5. elaeagnus mollis bud multiplication according to claim 1 and plant regeneration method, it is characterised in that：In step (1),
After rinsing 1h with a small amount of liquid detergent and 84 medicining liquid dipping 10min and flowing water, 40s is impregnated with 70% alcohol in super-clean bench, then
20min, aseptic water washing 4~5 times are impregnated with appropriate 0.1% mercuric chloride and a small amount of Tween~20, oscillation.
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| CN109804926B (en) * | 2019-02-14 | 2023-01-06 | 西安同人农林实业股份有限公司 | Elaeagnus mollis tissue culture medium and preparation method thereof |
| CN109964818B (en) * | 2019-04-23 | 2022-06-17 | 西安艾默农林生物技术有限公司 | Sterile Elaeagnus mollis oil seedling GA3Rooting medium and preparation method thereof |
| CN109937882B (en) * | 2019-04-23 | 2022-07-12 | 西安艾默农林生物技术有限公司 | Elaeagnus mollis tissue browning prevention culture medium and preparation method thereof |
| CN110235782A (en) * | 2019-06-17 | 2019-09-17 | 西安同人五凤农业有限公司 | A kind of production method of elaeagnus mollis artificial seed |
| CN111990253A (en) * | 2020-07-17 | 2020-11-27 | 上海培林生物科技有限公司 | Method for tissue culture of elaeagnus plants by using stem segments |
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