CN105766637B - The method for tissue culture of hemerocailis middendorffi " lace shawl " - Google Patents
The method for tissue culture of hemerocailis middendorffi " lace shawl " Download PDFInfo
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- 238000000034 method Methods 0.000 title claims abstract description 30
- 206010020649 Hyperkeratosis Diseases 0.000 claims abstract description 27
- 230000006698 induction Effects 0.000 claims abstract description 22
- 230000003716 rejuvenation Effects 0.000 claims abstract description 10
- 239000000463 material Substances 0.000 claims abstract description 6
- 230000004069 differentiation Effects 0.000 claims abstract description 5
- 239000001963 growth medium Substances 0.000 claims description 24
- 238000005286 illumination Methods 0.000 claims description 18
- 239000003415 peat Substances 0.000 claims description 9
- 235000019362 perlite Nutrition 0.000 claims description 9
- 239000010451 perlite Substances 0.000 claims description 9
- 239000011159 matrix material Substances 0.000 claims description 7
- 150000003839 salts Chemical class 0.000 claims description 6
- 229920001817 Agar Polymers 0.000 claims description 5
- TWFZGCMQGLPBSX-UHFFFAOYSA-N Carbendazim Natural products C1=CC=C2NC(NC(=O)OC)=NC2=C1 TWFZGCMQGLPBSX-UHFFFAOYSA-N 0.000 claims description 5
- 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 5
- 229930006000 Sucrose Natural products 0.000 claims description 5
- 239000008272 agar Substances 0.000 claims description 5
- JNPZQRQPIHJYNM-UHFFFAOYSA-N carbendazim Chemical compound C1=C[CH]C2=NC(NC(=O)OC)=NC2=C1 JNPZQRQPIHJYNM-UHFFFAOYSA-N 0.000 claims description 5
- 239000006013 carbendazim Substances 0.000 claims description 5
- 238000004140 cleaning Methods 0.000 claims description 5
- 239000002985 plastic film Substances 0.000 claims description 5
- 229920006255 plastic film Polymers 0.000 claims description 5
- 230000000384 rearing effect Effects 0.000 claims description 5
- 238000012549 training Methods 0.000 claims description 5
- 230000001939 inductive effect Effects 0.000 claims 2
- 241000409198 Packera aurea Species 0.000 claims 1
- 238000009395 breeding Methods 0.000 abstract description 5
- 230000001488 breeding effect Effects 0.000 abstract description 5
- 239000012882 rooting medium Substances 0.000 abstract description 2
- 230000004083 survival effect Effects 0.000 abstract description 2
- 238000012360 testing method Methods 0.000 abstract description 2
- 238000002054 transplantation Methods 0.000 abstract description 2
- 239000002585 base Substances 0.000 description 14
- 240000009206 Hemerocallis fulva Species 0.000 description 8
- 235000002941 Hemerocallis fulva Nutrition 0.000 description 8
- 241000196324 Embryophyta Species 0.000 description 5
- 239000002609 medium Substances 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 1
- 241000756137 Hemerocallis Species 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 241000234280 Liliaceae Species 0.000 description 1
- 244000018633 Prunus armeniaca Species 0.000 description 1
- 235000009827 Prunus armeniaca Nutrition 0.000 description 1
- 208000036142 Viral infection Diseases 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 235000012730 carminic acid Nutrition 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 229960002523 mercuric chloride Drugs 0.000 description 1
- LWJROJCJINYWOX-UHFFFAOYSA-L mercury dichloride Chemical compound Cl[Hg]Cl LWJROJCJINYWOX-UHFFFAOYSA-L 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
Classifications
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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
-
- 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
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
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- 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 the method for tissue culture of hemerocailis middendorffi " lace shawl ", this method comprises the following steps：(1) acquisition of sterilizable material；(2) callus induces；(3) bud breaks up；(4) rejuvenation culture；(5) culture of rootage；(6) hardening and transplanting.The present invention is using hemerocailis middendorffi " lace shawl " stem apex as explant, sterilized by explant and optimize callus induction, bud differentiation, rejuvenation, prescription of rooting medium and hardening condition of culture, induced by callus, bud differentiation, rejuvenation, culture of rootage and test tube transplantation of seedlings stage, realize the quick breeding of hemerocailis middendorffi " lace shawl " seedling, tissue-cultured seedling transplanting survival rate is up to more than 98%, for hemerocailis middendorffi is quickly bred, popularization and application lay the foundation.
Description
Technical field
The present invention relates to biological technical field, and in particular to the method for tissue culture of hemerocailis middendorffi " lace shawl ".
Background technology
Hemerocailis middendorffi, Liliaceae hemerocallis herbaceos perennial, the big color of tawny daylily numerous summer in few flower bloom, had
A variety of patterns such as yellow, apricot yellow, orange, carmine and purple, the florescence is long, flower amount is big, at the same have stronger drought resisting, cold-resistant,
The characteristics such as salt resistance alkali, disease and insect resistance, resistance to half shade.Tawny daylily is a kind of important ornamental plant in American-European countries, is that kind is most abundant
One of Perennial Flowers.Research and utilization work starting of the China to tawny daylily is than later, the only " Jin Wa of success popularization and application at present
Baby " is a kind of, and the tawny daylily kind overwhelming majority of country's plantation is high quoted from foreign countries, the few valency of these kind amounts.Tawny daylily setting percentage is low, and
Seed broadcasts seedling growth performance very different, and variety stability is low not to use seminal propagation typically, but uses tiller vegetative propagation side
Formula is bred.But because its speed is slow, general one plant of hemerocailis middendorffi, which is often only, can breed 3~4 plants, and individual plants are often only numerous
1~2 plant is grown, thus is difficult in adapt to the demand of market commercialisation production, causes the high present situation of the few valency of current excellent tawny daylily kind amount,
Seriously limit being widely applied for domestic tawny daylily new varieties.Method for tissue culture breeding is short with the breeding cycle, and whole year can
Bred, the features such as breeding coefficient is high, some virosis can also be eliminated, be solve the problems, such as tawny daylily amount reproduction most effective
Approach.
The content of the invention
It is an object of the invention to：For above-mentioned technical problem present in prior art, there is provided a kind of hemerocailis middendorffi " flower bud
The method for tissue culture of silk shawl ".
The present invention is achieved by the following technical solutions：
The method for tissue culture of hemerocailis middendorffi " lace shawl ", this method comprise the following steps：
(1) acquisition of sterilizable material, desirable hemerocailis middendorffi " lace shawl " stem apex, is cut into after cleaning up throughout the year
3 fritter 1cm, soaked 16 hours with the type solution of 4-5% mountains agriculture No.1 I, soak adds the salt of 1/4MS minimal mediums, base
Basal culture medium uses MS minimal mediums, wherein white sugar 20g/L, agar 5g/L, PH5.8, callus induction training is seeded in after taking-up
Support on base；
(2) callus induces：Under the conditions of 25 ± 2 DEG C, 1500-2500Lx of light intensity, illumination 12h/d, stem apex is visible after 40 days
Obvious callus, light culture 20-30 days；
(3) bud breaks up：Callus is connected on bud induction culture medium, at 25 ± 2 DEG C, light intensity 1500-
Under the conditions of 2500Lx, illumination 12h/d, culture starts bud after 25 days and is differentiated to form Multiple Buds；
(4) rejuvenation culture：Multiple Buds are transferred on strong seedling culture base, at 25 ± 2 DEG C, 1500-2500Lx of light intensity, light
Under the conditions of 12h/d, 20-30 days are cultivated to more than height of seedling 5cm；
(5) culture of rootage：Strong sprout is transferred on root media, at 25 ± 2 DEG C, 1500-2500Lx of light intensity, illumination
Under the conditions of 12h/d, base portion starts long root system after cultivating 15 days；
(6) hardening and transplanting：
1. hardening：The tissue-cultured seedling band bottle of long up to 10cm or so, long root >=5 piece is moved in common greenhouse, places 3-5
Bottle cap is opened after it, at 25-28 DEG C, humidity 70-80%, under the conditions of 7000-8000Lx of light intensity, hardening 3-5 days；
2. transplanting and culture：Tissue-cultured seedling after hardening is cleaned into root culture medium, soaks root with 0.1% carbendazim solution
After 30-60 seconds, transplant into the mixed-matrix of peat and perlite, with covered rearing with plastic film and shade net is covered, in 25-30
DEG C, humidity 80-95%, under the conditions of strong after light intensity is preferably first weak, cultivate 1 week, after be gradually opened film；Enter conventional pipe after 2 weeks
Reason, culture 1-2 months after, seedling.
Preferably, the calli induction media is the type 0.1%+6-BA1.8 of MS+ mountains agriculture No.1 II-2.3mg/L+
NAA0.3—0.6mg/L。
Preferably, the bud induction culture medium is the type 0.1%+6-BA3.5 of MS+ mountains agriculture No.1 II-4.5mg/L
+NAA0.18—0.2mg/L。
Preferably, the root media is the type 0.1%+IBA0.15-0.2mg/L+ of 1/2MS+ mountains agriculture No.1 II
NAA0.05—0.1mg/L。
Preferably, the strong seedling culture base is the type 0.1%+6-BA0.3mg/L+NAA0.05mg/ of MS+ mountains agriculture No.1 II
L。
Preferably, the calli induction media is the type 0.1%+6-BA2.0mg/L+ of MS+ mountains agriculture No.1 II
NAA0.5mg/L。
Preferably, the bud induction culture medium is the type 0.1%+6-BA4.0mg/L+ of MS+ mountains agriculture No.1 II
NAA0.2mg/L。
Preferably, the root media is the type 0.1%+IBA0.2mg/L+NAA0.1mg/ of 1/2MS+ mountains agriculture No.1 II
L。
Preferably, in the mixed-matrix of the peat and perlite, the volume ratio of peat and perlite is 7: 3.
In summary, by adopting the above-described technical solution, the beneficial effects of the invention are as follows：
1st, the present invention is sterilized by explant using hemerocailis middendorffi " lace shawl " stem apex as explant and optimization callus lures
Lead, bud differentiation, rejuvenation, prescription of rooting medium and hardening condition of culture, induced by callus, bud differentiation, rejuvenation, culture of rootage
And the test tube transplantation of seedlings stage, realize the quick breeding of hemerocailis middendorffi " lace shawl " seedling, tissue-cultured seedling transplanting survival rate is up to 98%
More than, for hemerocailis middendorffi is quickly bred, popularization and application lay the foundation.
2nd, it is of the invention compared with existing method for tissue culture, from 6-BA, and do not have to KT, because 6-BA can cause
Callus is grown good, easily sprouts, and agriculture No.1 in mountain is added inside culture medium, and hemerocailis middendorffi tissue cultures whole process is effectively ensured
It is pollution-free, compared to existing alcohol, mercuric chloride sterilize etc. disinfection way, with mountain agriculture No.1 can obtain detoxic seedling have 90% with
On, considerably beyond other disinfection way.
Embodiment
All features disclosed in this specification, or disclosed all methods or during the step of, except mutually exclusive
Feature and/or step beyond, can combine in any way.
Any feature disclosed in this specification (including any accessory claim, summary), unless specifically stated otherwise,
Replaced by other equivalent or with similar purpose alternative features.I.e., unless specifically stated otherwise, each feature is a series of
An example in equivalent or similar characteristics.
Embodiment 1
The method for tissue culture of hemerocailis middendorffi " lace shawl ", this method comprise the following steps：
(1) acquisition of sterilizable material, desirable hemerocailis middendorffi " lace shawl " stem apex, is cut into after cleaning up throughout the year
3 fritter 1cm, soaked 16 hours with the type solution of 4-5% mountains agriculture No.1 I, soak adds the salt of 1/4MS minimal mediums, MS
Minimal medium uses white sugar 20g/L, agar 5g/L, PH5.8, is seeded in after taking-up on calli induction media, callus induction
Culture medium is the type 0.1%+6-BA1.8mg/L+NAA0.3mg/L of MS+ mountains agriculture No.1 II；
(2) callus induces：Under the conditions of 25 ± 2 DEG C, light intensity 1500Lx, illumination 12h/d, stem apex is visible obvious after 40 days
Callus, light culture 20 days；
(3) bud breaks up：Callus is connected on bud induction culture medium, bud induction culture medium is the agriculture of MS+ mountains
Type 0.1%+6-the BA3.5mg/L+NAA0.18mg/L of No.1 II, under the conditions of 25 ± 2 DEG C, light intensity 1500Lx, illumination 12h/d,
Culture starts bud after 25 days and is differentiated to form Multiple Buds；
(4) rejuvenation culture：Multiple Buds are transferred on strong seedling culture base, strong seedling culture base is the type of MS+ mountains agriculture No.1 II
0.1%+6-BA0.3mg/L+NAA0.05mg/L, under the conditions of 25 ± 2 DEG C, light intensity 1500Lx, illumination 12h/d, cultivate 20 days
To more than height of seedling 5cm；
(5) culture of rootage：Strong sprout is transferred on root media, root media is the type of 1/2MS+ mountains agriculture No.1 II
0.1%+IBA0.15mg/L+NAA0.05mg/L, under the conditions of 25 ± 2 DEG C, light intensity 1500Lx, illumination 12h/d, after cultivating 15 days
Base portion starts long root system；
(6) hardening and transplanting：
1. hardening：The tissue-cultured seedling band bottle of long up to 10cm or so, long root >=5 piece is moved in common greenhouse, after placing 3 days
Bottle cap is opened, at 25-28 DEG C, humidity 70-80%, under the conditions of light intensity 7000Lx, hardening 3 days；
2. transplanting and culture：Tissue-cultured seedling after hardening is cleaned into root culture medium, soaks root with 0.1% carbendazim solution
After 30 seconds, transplant in the mixed-matrix for being 7: 3 to the volume ratio of peat and perlite, sheltered from heat or light with covered rearing with plastic film and covering
Net, at 25-30 DEG C, humidity 80-95%, under the conditions of strong after light intensity is preferably first weak, cultivate 1 week, after be gradually opened film；After 2 weeks
Into Routine Management, after cultivating January, seedling.
Embodiment 2
The method for tissue culture of hemerocailis middendorffi " lace shawl ", this method comprise the following steps：
(1) acquisition of sterilizable material, desirable hemerocailis middendorffi " lace shawl " stem apex, is cut into after cleaning up throughout the year
3 fritter 1cm, soaked 16 hours with the type solution of 4-5% mountains agriculture No.1 I, soak adds the salt of 1/4MS minimal mediums, MS
Minimal medium uses white sugar 20g/L, agar 5g/L, PH5.8, is seeded in after taking-up on calli induction media, callus induction
Culture medium is the type 0.1%+6-BA 2.0mg/L+NAA0.5mg/L of MS+ mountains agriculture No.1 II；
(2) callus induces：Under the conditions of 25 ± 2 DEG C, light intensity 2000Lx, illumination 12h/d, stem apex is visible obvious after 40 days
Callus, light culture 25 days；
(3) bud breaks up：Callus is connected on bud induction culture medium, bud induction culture medium is the agriculture of MS+ mountains
Type 0.1%+6-BA the 4.0mg/L+NAA0.2mg/L of No.1 II, under the conditions of 25 ± 2 DEG C, light intensity 2000Lx, illumination 12h/d,
Culture starts bud after 25 days and is differentiated to form Multiple Buds；
(4) rejuvenation culture：Multiple Buds are transferred on strong seedling culture base, strong seedling culture base is the type of MS+ mountains agriculture No.1 II
0.1%+6-BA 0.3mg/L+NAA0.05mg/L, under the conditions of 25 ± 2 DEG C, light intensity 2000Lx, illumination 12h/d, cultivate 25 days
To more than height of seedling 5cm；
(5) culture of rootage：Strong sprout is transferred on root media, root media is the type of 1/2MS+ mountains agriculture No.1 II
0.1%+IBA 0.2mg/L+NAA0.1mg/L, under the conditions of 25 ± 2 DEG C, light intensity 2000Lx, illumination 12h/d, after cultivating 15 days
Base portion starts long root system；
(6) hardening and transplanting：
1. hardening：The tissue-cultured seedling band bottle of long up to 10cm or so, long root >=5 piece is moved in common greenhouse, after placing 4 days
Bottle cap is opened, at 25-28 DEG C, humidity 70-80%, under the conditions of light intensity 7500Lx, hardening 4 days；
2. transplanting and culture：Tissue-cultured seedling after hardening is cleaned into root culture medium, soaks root with 0.1% carbendazim solution
After 45 seconds, transplant in the mixed-matrix for being 7: 3 to the volume ratio of peat and perlite, sheltered from heat or light with covered rearing with plastic film and covering
Net, at 25-30 DEG C, humidity 80-95%, under the conditions of strong after light intensity is preferably first weak, cultivate 1 week, after be gradually opened film；After 2 weeks
Into Routine Management, after cultivating 1 first quarter moon, seedling.
Embodiment 3
The method for tissue culture of hemerocailis middendorffi " lace shawl ", this method comprise the following steps：
(1) acquisition of sterilizable material, desirable hemerocailis middendorffi " lace shawl " stem apex, is cut into after cleaning up throughout the year
3 fritter 1cm, soaked 16 hours with the type solution of 4-5% mountains agriculture No.1 I, soak adds the salt of 1/4MS minimal mediums, MS
Minimal medium uses white sugar 20g/L, agar 5g/L, PH5.8, is seeded in after taking-up on calli induction media, callus induction
Culture medium is the type 0.1%+6-BA2.3mg/L+NAA0.6mg/L of MS+ mountains agriculture No.1 II；
(2) callus induces：Under the conditions of 25 ± 2 DEG C, light intensity 2500Lx, illumination 12h/d, stem apex is visible obvious after 40 days
Callus, light culture 30 days；
(3) bud breaks up：Callus is connected on bud induction culture medium, bud induction culture medium is the agriculture of MS+ mountains
Type 0.1%+6-the BA4.5mg/L+NAA0.2mg/L of No.1 II, under the conditions of 25 ± 2 DEG C, light intensity 2500Lx, illumination 12h/d, training
Start bud after supporting 25 days and be differentiated to form Multiple Buds；
(4) rejuvenation culture：Multiple Buds are transferred on strong seedling culture base, strong seedling culture base is the type of MS+ mountains agriculture No.1 II
0.1%+6-BA 0.3mg/L+NAA0.05mg/L, under the conditions of 25 ± 2 DEG C, light intensity 2500Lx, illumination 12h/d, cultivate 30 days
To more than height of seedling 5cm；
(5) culture of rootage：Strong sprout is transferred on root media, root media is the type of 1/2MS+ mountains agriculture No.1 II
0.1%+IBA0.2mg/L+NAA0.1mg/L, under the conditions of 25 ± 2 DEG C, light intensity 2500Lx, illumination 12h/d, base after cultivating 15 days
Portion starts long root system；
(6) hardening and transplanting：
1. hardening：The tissue-cultured seedling band bottle of long up to 10cm or so, long root >=5 piece is moved in common greenhouse, after placing 5 days
Bottle cap is opened, at 25-28 DEG C, humidity 70-80%, under the conditions of light intensity 8000Lx, hardening 5 days；
2. transplanting and culture：Tissue-cultured seedling after hardening is cleaned into root culture medium, soaks root with 0.1% carbendazim solution
After 60 seconds, transplant in the mixed-matrix for being 7: 3 to the volume ratio of peat and perlite, sheltered from heat or light with covered rearing with plastic film and covering
Net, at 25-30 DEG C, humidity 80-95%, under the conditions of strong after light intensity is preferably first weak, cultivate 1 week, after be gradually opened film；After 2 weeks
Into Routine Management, after culture 2 months, seedling.
Particular embodiments described above, the purpose of the present invention, technical scheme and beneficial effect are carried out further in detail
Describe in detail it is bright, should be understood that the foregoing is only the present invention specific embodiment, be not intended to limit the invention.This
Invention expands to any new feature disclosed in this manual or any new combination, and any new method for disclosing or
The step of process or any new combination.
Claims (9)
1. the method for tissue culture of hemerocailis middendorffi " lace shawl ", it is characterised in that this method comprises the following steps：
（1）The acquisition of sterilizable material, desirable hemerocailis middendorffi " lace shawl " stem apex, it is small to be cut into 3 after cleaning up throughout the year
Block 1cm, soaked 16 hours with the type solution of 4-5% mountains agriculture No.1 I, soak adds the salt of 1/4 MS minimal mediums, base
Basal culture medium uses MS minimal mediums, wherein white sugar 20g/L, agar 5g/L, PH5.8, callus induction training is seeded in after taking-up
Support on base；
（2）Callus induces：Under the conditions of 25 ± 2 DEG C, the Lx of light intensity 1500-2500, illumination 12h/d, stem apex is visible bright after 40 days
Aobvious callus, light culture 20-30 days；
（3）Bud breaks up：Callus is connected on bud induction culture medium, at 25 ± 2 DEG C, the Lx of light intensity 1500-2500, light
Under the conditions of 12h/d, culture starts bud after 25 days and is differentiated to form Multiple Buds；
（4）Rejuvenation culture：Multiple Buds are transferred on strong seedling culture base, at 25 ± 2 DEG C, 1500-2500Lx of light intensity, illumination
Under the conditions of 12h/d, 20-30 days are cultivated to more than height of seedling 5cm；
（5）Culture of rootage：Strong sprout is transferred on root media, at 25 ± 2 DEG C, 1500-2500Lx of light intensity, illumination 12h/d
Under the conditions of, base portion starts long root system after cultivating 15 days；
（6）Hardening and transplanting：
1. hardening：The tissue-cultured seedling band bottle of long up to 10cm, long root >=5 piece is moved in common greenhouse, opened after placing 3-5 days
Bottle cap, at 25-28 DEG C, humidity 70-80%, under the conditions of 7000-8000Lx of light intensity, hardening 3-5 days；
Transplanting and culture：Tissue-cultured seedling after hardening is cleaned into root culture medium, soaks root 30-60 with 0.1% carbendazim solution
After second, transplant into the mixed-matrix of peat and perlite, with covered rearing with plastic film and cover shade net, it is wet at 25-30 DEG C
Degree 80-95%, under the conditions of strong after light intensity is preferably first weak, cultivate 1 week, after be gradually opened film；Enter Routine Management after 2 weeks, cultivate
After 1-2 months, seedling.
2. the method for tissue culture of hemerocailis middendorffi " lace shawl " as claimed in claim 1, it is characterised in that the callus lures
It is the type 0.1%+6-BA1.8 of MS+ mountains agriculture No.1 II-2.3mg/L+NAA0.3-0.6mg/L to lead culture medium.
3. the method for tissue culture of hemerocailis middendorffi " lace shawl " as claimed in claim 1, it is characterised in that the bud differentiation
Inducing culture is the type 0.1%+6-BA3.5 of MS+ mountains agriculture No.1 II-4.5mg/L+NAA0.18-0.2mg/L.
4. the method for tissue culture of hemerocailis middendorffi " lace shawl " as claimed in claim 1, it is characterised in that the training of taking root
It is the type 0.1%+IBA0.15-0.2mg/L+NAA0.05 of 1/2MS+ mountains agriculture No.1 II-0.1mg/L to support base.
5. the method for tissue culture of hemerocailis middendorffi " lace shawl " as claimed in claim 1, it is characterised in that the strong sprout training
It is the type 0.1%+6-BA 0.3mg/L+NAA0.05mg/L of MS+ mountains agriculture No.1 II to support base.
6. the method for tissue culture of hemerocailis middendorffi " lace shawl " as claimed in claim 1 or 2, it is characterised in that described to be cured
It is the type 0.1%+6-BA 2.0mg/L+NAA0.5mg/L of MS+ mountains agriculture No.1 II to hinder inducing culture.
7. the method for tissue culture of the hemerocailis middendorffi " lace shawl " as described in claim 1 or 3, it is characterised in that the bud
Induction culture medium is the type 0.1%+6-BA 4.0mg/L+NAA0.2mg/L of MS+ mountains agriculture No.1 II.
8. the method for tissue culture of the hemerocailis middendorffi " lace shawl " as described in claim 1 or 4, it is characterised in that the life
Root culture medium is the type 0.1%+IBA 0.2mg/L+NAA0.1mg/L of 1/2MS+ mountains agriculture No.1 II.
9. the method for tissue culture of hemerocailis middendorffi " lace shawl " as claimed in claim 1, it is characterised in that the peat with
In the mixed-matrix of perlite, the volume ratio of peat and perlite is 7: 3.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
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