CN107494264A - A kind of method of Pterospermi Heterophylli tissue culture outside sprout-cultivating-bottle radication - Google Patents
A kind of method of Pterospermi Heterophylli tissue culture outside sprout-cultivating-bottle radication Download PDFInfo
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- CN107494264A CN107494264A CN201710769397.4A CN201710769397A CN107494264A CN 107494264 A CN107494264 A CN 107494264A CN 201710769397 A CN201710769397 A CN 201710769397A CN 107494264 A CN107494264 A CN 107494264A
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- seedling
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- outside sprout
- pterospermi heterophylli
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- 210000001519 tissues Anatomy 0.000 title claims abstract description 28
- 239000011159 matrix material Substances 0.000 claims abstract description 17
- 230000003716 rejuvenation Effects 0.000 claims abstract description 13
- 230000000249 desinfective Effects 0.000 claims abstract description 6
- 239000002609 media Substances 0.000 claims description 25
- 239000003415 peat Substances 0.000 claims description 19
- 238000004659 sterilization and disinfection Methods 0.000 claims description 19
- 230000001954 sterilising Effects 0.000 claims description 14
- 239000010451 perlite Substances 0.000 claims description 9
- 235000019362 perlite Nutrition 0.000 claims description 9
- 239000010455 vermiculite Substances 0.000 claims description 9
- 235000019354 vermiculite Nutrition 0.000 claims description 9
- 229910052902 vermiculite Inorganic materials 0.000 claims description 9
- 239000003795 chemical substances by application Substances 0.000 claims description 8
- 239000000203 mixture Substances 0.000 claims description 7
- 239000004033 plastic Substances 0.000 claims description 7
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 claims description 6
- 239000006870 ms-medium Substances 0.000 claims description 6
- LBLSRDDHGILUJH-UHFFFAOYSA-N 4-(1H-indol-2-yl)butanoic acid Chemical compound C1=CC=C2NC(CCCC(=O)O)=CC2=C1 LBLSRDDHGILUJH-UHFFFAOYSA-N 0.000 claims description 5
- YGGXZTQSGNFKPJ-UHFFFAOYSA-N methyl 2-naphthalen-1-ylacetate Chemical compound C1=CC=C2C(CC(=O)OC)=CC=CC2=C1 YGGXZTQSGNFKPJ-UHFFFAOYSA-N 0.000 claims description 5
- 241000219430 Betula pendula Species 0.000 claims description 4
- DHMQDGOQFOQNFH-UHFFFAOYSA-N glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims description 4
- 238000005286 illumination Methods 0.000 claims description 4
- PVNIIMVLHYAWGP-UHFFFAOYSA-N nicotinic acid Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 claims description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 4
- 230000004069 differentiation Effects 0.000 claims description 3
- 238000007580 dry-mixing Methods 0.000 claims description 3
- 239000000835 fiber Substances 0.000 claims description 3
- NPDODHDPVPPRDJ-UHFFFAOYSA-N permanganate Chemical compound [O-][Mn](=O)(=O)=O NPDODHDPVPPRDJ-UHFFFAOYSA-N 0.000 claims description 3
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Inorganic materials [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 claims description 3
- ARUVKPQLZAKDPS-UHFFFAOYSA-L Copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 claims description 2
- 239000004471 Glycine Substances 0.000 claims description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N HCl Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 2
- 239000007836 KH2PO4 Substances 0.000 claims description 2
- SQQMAOCOWKFBNP-UHFFFAOYSA-L Manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 2
- 229910004619 Na2MoO4 Inorganic materials 0.000 claims description 2
- 231100000614 Poison Toxicity 0.000 claims description 2
- TVXXNOYZHKPKGW-UHFFFAOYSA-N Sodium molybdate Chemical compound [Na+].[Na+].[O-][Mo]([O-])(=O)=O TVXXNOYZHKPKGW-UHFFFAOYSA-N 0.000 claims description 2
- 229960000344 Thiamine hydrochloride Drugs 0.000 claims description 2
- NWONKYPBYAMBJT-UHFFFAOYSA-L Zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 claims description 2
- 239000001110 calcium chloride Substances 0.000 claims description 2
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 2
- 229910052927 chalcanthite Inorganic materials 0.000 claims description 2
- ZGTMUACCHSMWAC-UHFFFAOYSA-L disodium;2-[2-[carboxylatomethyl(carboxymethyl)amino]ethyl-(carboxymethyl)amino]acetate Chemical compound [Na+].[Na+].OC(=O)CN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O ZGTMUACCHSMWAC-UHFFFAOYSA-L 0.000 claims description 2
- 229910052564 epsomite Inorganic materials 0.000 claims description 2
- 229910000357 manganese(II) sulfate Inorganic materials 0.000 claims description 2
- 229910052603 melanterite Inorganic materials 0.000 claims description 2
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 2
- 235000001968 nicotinic acid Nutrition 0.000 claims description 2
- 229960003512 nicotinic acid Drugs 0.000 claims description 2
- 239000011664 nicotinic acid Substances 0.000 claims description 2
- 239000002574 poison Substances 0.000 claims description 2
- 239000011684 sodium molybdate Substances 0.000 claims description 2
- 235000019190 thiamine hydrochloride Nutrition 0.000 claims description 2
- 239000011747 thiamine hydrochloride Substances 0.000 claims description 2
- DPJRMOMPQZCRJU-UHFFFAOYSA-M thiamine hydrochloride Chemical compound Cl.[Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N DPJRMOMPQZCRJU-UHFFFAOYSA-M 0.000 claims description 2
- 229910000368 zinc sulfate Inorganic materials 0.000 claims description 2
- 239000011686 zinc sulphate Substances 0.000 claims description 2
- 241000208140 Acer Species 0.000 claims 1
- KGBXLFKZBHKPEV-UHFFFAOYSA-N Boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 claims 1
- 229940064880 Inositol 100 MG Drugs 0.000 claims 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M Monopotassium phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims 1
- 235000006508 Nelumbo nucifera Nutrition 0.000 claims 1
- 240000002853 Nelumbo nucifera Species 0.000 claims 1
- 235000006510 Nelumbo pentapetala Nutrition 0.000 claims 1
- 241000409198 Packera aurea Species 0.000 claims 1
- 230000004083 survival Effects 0.000 abstract description 9
- 238000005516 engineering process Methods 0.000 abstract description 7
- 238000004519 manufacturing process Methods 0.000 abstract description 4
- 238000000034 method Methods 0.000 abstract description 4
- 230000003750 conditioning Effects 0.000 abstract description 2
- 241000196324 Embryophyta Species 0.000 description 5
- 210000004027 cells Anatomy 0.000 description 3
- 241000190047 Altingia Species 0.000 description 2
- 241000142952 Hamamelidaceae Species 0.000 description 2
- VZJVWSHVAAUDKD-UHFFFAOYSA-N Potassium permanganate Chemical compound [K+].[O-][Mn](=O)(=O)=O VZJVWSHVAAUDKD-UHFFFAOYSA-N 0.000 description 2
- 230000035699 permeability Effects 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 241000195493 Cryptophyta Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 206010019468 Hemiplegia Diseases 0.000 description 1
- 206010022114 Injury Diseases 0.000 description 1
- 241000208682 Liquidambar Species 0.000 description 1
- 235000006550 Liquidambar Nutrition 0.000 description 1
- 208000002193 Pain Diseases 0.000 description 1
- 210000002435 Tendons Anatomy 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 230000003213 activating Effects 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-N ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 1
- 230000000844 anti-bacterial Effects 0.000 description 1
- 230000001488 breeding Effects 0.000 description 1
- 230000001684 chronic Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 239000003595 mist Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000006011 modification reaction Methods 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 210000000056 organs Anatomy 0.000 description 1
- 230000001151 other effect Effects 0.000 description 1
- 239000002985 plastic film Substances 0.000 description 1
- 230000001902 propagating Effects 0.000 description 1
- 210000001938 protoplasts Anatomy 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 230000002040 relaxant effect Effects 0.000 description 1
- 201000003068 rheumatic fever Diseases 0.000 description 1
- 230000002786 root growth Effects 0.000 description 1
- 238000010008 shearing Methods 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 230000033772 system development Effects 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
- A01H4/001—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
-
- 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
Abstract
The invention discloses a kind of method of Pterospermi Heterophylli tissue culture outside sprout-cultivating-bottle radication, comprise the following steps：Tissue-cultured seedling rejuvenation, outside sprout-cultivating-bottle and seedling management；The present invention is by tissue culture rejuvenation technique, the selection of outside sprout-cultivating-bottle container, matrix is selected with disinfecting, outside sprout-cultivating-bottle technology, seedling stage environment conditioning are taken root in production applied to Pterospermi Heterophylli tissue-cultured seedling, the rooting rate of obtained Pterospermi Heterophylli tissue culture outside sprout-cultivating-bottle radication is up to 90%, survival rate 95%.The present invention not only increases the transplanting survival rate of Pterospermi Heterophylli tissue culture rooted seedling, reduces cost, shortens growing-seedling period, and provide a novel technical method for Pterospermi Heterophylli tissue culture seedling rooting.
Description
Technical field
The present invention relates to tissue culture outside sprout-cultivating-bottle radication field, more particularly, to a kind of side of Pterospermi Heterophylli tissue culture outside sprout-cultivating-bottle radication
Method.
Background technology
Pterospermi Heterophylli Altingia chingii Mete are Hamamelidaceae aiphylliums, Chinese Second Class Key Protected Plant.It is Chinese special
There is kind, there are the Comprehensive Traits between Liquidambar Liguidambar and Altingia Altingia two category, to studying Hamamelidaceae system
Development has scientific value.Wood quality is excellent, and rotation plane property is good, can make rotation plane product.Bark has relaxing tendons and activating collaterals, wind-dispelling to remove simultaneously
It is wet and other effects among the people for treating the diseases such as rheumatic arthritis, chronic lumbocrural pain, hemiplegia, traumatic injury, it is evident in efficacy.Cause
Natural stands is serious and natural propagation is difficult by artificial disturbance, and distribution is more and more narrow, causes wild resource extremely rare.Plant
The tissue cultures of thing, which refer to, isolates the tissue to suit the requirements, organ or cell, protoplast etc. from plant, passes through sterile behaviour
Make, cultivated under manual control condition to obtain other products of the intact plant of regeneration or production with economic value
Technology.Pterospermi Heterophylli tissue culture technology is gradually grown up, and advantage is created for the seeds popularization and application.
But relative to cuttage and seedling culture, tissue-cultured seedling has that culture program is more, transplanting survival rate is not high, seedling cost is high etc. asks
Topic.Therefore, on the basis of Pterospermi Heterophylli group culturation rapid propagating technology maturation, the rejuvenation of Pterospermi Heterophylli subculture sprout, outside sprout-cultivating-bottle technology are carried out
Research, to reduce tissue-cultured seedling culture of rootage link, shorten growing-seedling period, reduce seedling cost, Pterospermi Heterophylli tissue culture rooted seedling is big
Field transplanting is simplified, and further improves Pterospermi Heterophylli tissue-culturing rapid propagation and industrial breeding technique system, promotes the popularization of tissue-cultured seedling should
With having important practical significance.
The content of the invention
It is an object of the invention to provide a kind of rooting rate up to more than 90%, it is not required to carry out hardening, can be directly transplanted to
In nutrient bag, the method for the Pterospermi Heterophylli tissue culture outside sprout-cultivating-bottle radication of survival rate more than 95%.
In order to realize the object of the invention, the invention provides a kind of method of Pterospermi Heterophylli tissue culture outside sprout-cultivating-bottle radication, including such as
Lower step：
Tissue-cultured seedling rejuvenation：Fiber differentiation bud clump in Pterospermi Heterophylli tissue-cultured seedling access modified MS medium is long sturdy, answered
Strong bottle seedling；
Outside sprout-cultivating-bottle：Seedling medium after sterilization is put into and taken root in container, seedling medium is highly 2cm, by nursery base
Matter is floating, compacting, is drenched with clear water, and the deep apertures of multiple 1.0-1.5cm are stabbed out on seedling medium, and selection height is 2cm-
3cm strengthens the rejuvenation bottle seedling of bud, cuts strong bud, strengthens the fast quickly dipped upper root-growing agent of bud, is then inserted on the aperture of seedling medium, buckles
Take root container lid；
Seedling management：The container of taking root for being inserted with seedling in outside sprout-cultivating-bottle step is moved on to culturing room to cultivate 15 days, temperature control
For system at 26 ± 2 DEG C, light culture opens light, daily illumination 10h after 2-3 days；Move on to hardening canopy within 16th day, continue to take root, refine
Seedling, hardening canopy temperature are 15-30 DEG C, humidity >=70%；The 16-18 days, at 10 points in the morning, interior external sunshade was complete to 4 points or so of afternoon
Deploy in portion；The 19-22 days, at 10 points in the morning deployed to 4 points or so of afternoon, external sunshade, afterwards full exposure hardening, until half is wooden
Change, the thickening greening of blade, you can field planting or transplanting.
Further, in the tissue-cultured seedling tissue-cultured seedling rejuvenation, modified MS medium is：NH4NO3500mg/L, KNO3
1900mg/L, MgSO4·7H2O 370mg/L, KH2PO4225mg/L, CaCl2·2H2O 440mg/L, MnSO4·4H2O
16.9mg/L ZnSO4·7H2O 8.6mg/L, H3BO36.2mg/L, KI 0.83mg/L, Na2MoO4·2H2O 0.25mg/L,
CuSO4·5H2O 0.025mg/L, CoCl2·6H2O 0.025mg/L, FeSO4·7H2O 27.8mg/L, Na2-EDTA·2H2O
37.3mg/L, glycine 2.0mg/L, thiamine hydrochloride 2.0mg/L, puridoxine hydrochloride 0.5mg/L, IV B nicotinic acid 2.0mg/L, flesh
Alcohol 100mg/L.
Further, seedling medium described in outside sprout-cultivating-bottle step is the mixture of peat, vermiculite and perlite；It is preferred that
, the weight ratio of the peat, vermiculite and perlite is 3:1:1.
Further, the sterilization method of the seedling medium is：Peat is spread out to be exposed to the sun under the sun 2-3 days and sterilized, is disappeared
Peat pack after poison is standby, mixes the peat after sterilization, vermiculite and perlite in proportion before rooting and transplant, and dry mixing is uniform to be obtained
To mixed-matrix, mixed-matrix is carried out disinfection with 0.3% liquor potassic permanganate to mixed-matrix and moistened, the mixed base after sterilization
Matter at least vexed close one day produces seedling medium.
Further, container of being taken root described in outside sprout-cultivating-bottle step is plastic preserving box with a lid；Preferably, the modeling
The specification for expecting crisper is 20cm × 13cm × 12cm.
Further, root-growing agent is formed by indolebutyric acid and methyl α-naphthyl acetate mixture described in outside sprout-cultivating-bottle step；Preferably, institute
The mass ratio for stating indolebutyric acid and methyl α-naphthyl acetate is 3:2.
Further, root-growing agent described in outside sprout-cultivating-bottle step is 800ppm.
Further, the tissue-cultured seedling is Fujian Pterospermi Heterophylli tissue-cultured seedling or leaflet Pterospermi Heterophylli tissue-cultured seedling.
The present invention uses modified MS medium, reduces ammonia state N in formula, improves P content, promotes the hair of stem branch and root
Educate, induce and control bud clump is long slightly to become strong, be easy to smoothly complete outside sprout-cultivating-bottle in next step.
Present invention container of taking root uses plastic preserving box with a lid, and plastic material is light, transparent, ensure that sprout grows
Take root required humidity, illumination etc..The specification of plastic preserving box is 20cm × 13cm × 12cm, both can conveniently load matrix,
100-150 strain stalwartness budlets are inserted into again.
As taking root seedling medium, gas permeability and moisture retention are good for selection peat, vermiculite, the mixture of perlite, when peat,
The weight of vermiculite and perlite ratio is 3:1:When 1, transparent performance is good.Peat selects northeast peat or import peat, permeability good
It is good, it is superior in quality.Peat carries out being exposed to the sun 2-3 days under the sun, is carried out disinfection using sunshine, solar ultraviolet is to comprising thin
Bacterium, fungi, Richettsia, virus and algae etc. have good bactericidal effect.Matrix after sterilization, with 0.3% potassium permanganate
Mixed-matrix after solution disinfection is at least vexed to be closed one day, seedling medium is had a micro- fermentation growth course, can improve sterilization effect
Fruit and transplanting survival rate.It is vexed to close method to cover layer of plastic film on mixed-matrix after sterilization.
In seedling management step, light culture 2-3 days is the cell yellow in order that after the strong bud of shearing on wound, and then is converted
To break up the cell of root system, be advantageous to take root, and be beneficial to root growth.
Hardening canopy culture is that crisper is opened when temperature is less than 15 DEG C, closes surrounding windowing part.When humidity is less than
70%, micro- mist spray, increase humidity and water supply in media in box, be advantageous to seedling growth.
The present invention is by tissue culture rejuvenation technique, the selection of outside sprout-cultivating-bottle container, matrix selection and disinfects, outside sprout-cultivating-bottle skill
Art, seedling stage environment conditioning are taken root in production applied to Pterospermi Heterophylli tissue-cultured seedling, the life of obtained Pterospermi Heterophylli tissue culture outside sprout-cultivating-bottle radication
Root rate is up to 90%, survival rate 95%.The present invention not only increases the transplanting survival rate of Pterospermi Heterophylli tissue culture rooted seedling, reduces
Cost, growing-seedling period is shortened, and a novel technical method is provided for Pterospermi Heterophylli tissue culture seedling rooting.
Embodiment
Embodiments of the invention are described below in detail, the example of the embodiment is intended to be used to explain the present invention, and can not
It is interpreted as limitation of the present invention.In the examples where no specific technique or condition is specified, according to described by document in the art
Technology or condition or carried out according to product description.Agents useful for same or the unreceipted production firm person of instrument, are to lead to
Cross the conventional products of acquisition purchased in market.
Embodiment 1
Tissue-cultured seedling rejuvenation：Fiber differentiation bud clump in Pterospermi Heterophylli tissue-cultured seedling access modified MS medium is long sturdy, answered
Strong bottle seedling；
Prepare seedling medium：Peat is spread out to be exposed to the sun under the sun 2-3 days and sterilized, the peat pack after sterilization is standby, raw
Root transplanting before by the peat after sterilization, vermiculite and perlite in mass ratio be 3:1:1 mixing, dry mixing uniformly obtain mixed-matrix,
Mixed-matrix is carried out disinfection with 0.3% liquor potassic permanganate to mixed-matrix and moistened, the mixed-matrix after sterilization is vexed to be closed one day
Produce seedling medium；
Outside sprout-cultivating-bottle：Seedling medium after sterilization is put into and taken root in container, seedling medium is highly 2cm, by nursery base
Matter is floating, compacting, is drenched with clear water, and the deep apertures of multiple 1.0-1.5cm are stabbed out on seedling medium, and selection height is 2cm-
3cm strengthens the rejuvenation bottle seedling of bud, cuts strong bud, and the strong fast quickly dipped upper 800ppm of bud root-growing agent (presses matter by indolebutyric acid and methyl α-naphthyl acetate
Measure ratio 3:2 mixtures form), it is then inserted on the aperture of seedling medium, aperture is filled and led up, inserts 100 plants on seedling medium altogether
Strong bud, buckles container lid of taking root；The container of taking root is that specification is 20cm × 13cm × 12cm plastic preserving boxes with a lid；
Seedling management：The above-mentioned container of taking root for being inserted with seedling is moved on to culturing room to cultivate 15 days, temperature control is 26 ± 2
DEG C, light culture opens light, daily illumination 10h after 2-3 days；Move on to hardening canopy within 16th day, continue to take root, hardening, hardening canopy temperature
Spend for 15-30 DEG C, humidity >=70%；The 16-18 days, at 10 points in the morning all deployed to 4 points or so of afternoon, interior external sunshade；19-
22 days, at 10 points in the morning deployed to 4 points or so of afternoon, external sunshade, afterwards full exposure hardening, until semi-lignified, the thickening change of blade
It is green, you can field planting or transplanting, to count the rooting rate and survival rate of seedling.
Test Pterospermi Heterophylli seedling has 95 contain more than 3 root systems, 90 Pterospermi Heterophylli seedlings grow fine, and 10 blades become
Yellow or withered, the rooting rate of the Pterospermi Heterophylli tissue culture outside sprout-cultivating-bottle radication obtained in summary using technical solution of the present invention is reachable
90%, survival rate 95%.
Although embodiments of the invention have been shown and described above, it is to be understood that above-described embodiment is example
Property, it is impossible to limitation of the present invention is interpreted as, one of ordinary skill in the art is not departing from the principle and objective of the present invention
In the case of above-described embodiment can be changed within the scope of the invention, change, replace and modification.
Claims (8)
- A kind of 1. method of Pterospermi Heterophylli tissue culture outside sprout-cultivating-bottle radication, it is characterised in that comprise the following steps：Tissue-cultured seedling rejuvenation：Fiber differentiation bud clump in Pterospermi Heterophylli tissue-cultured seedling access modified MS medium is long sturdy, obtain rejuvenation bottle Seedling；Outside sprout-cultivating-bottle：Seedling medium after sterilization is put into and taken root in container, seedling medium is highly 2cm, and seedling medium is smeared Flat, compacting, is drenched with clear water, and the deep apertures of multiple 1.0-1.5cm are stabbed out on seedling medium, and selection height is strengthened for 2cm-3cm The rejuvenation bottle seedling of bud, after cutting strong bud, the fast quickly dipped upper root-growing agent of bud is strengthened, is then inserted on the aperture of seedling medium, buckles life Root container lid；Seedling management：The above-mentioned container of taking root for being inserted with seedling is moved on to culturing room to cultivate 15 days, temperature control is at 26 ± 2 DEG C, secretly After culture 2-3 days, light, daily illumination 10h are opened；Move on to hardening canopy within 16th day, continue to take root, hardening, hardening canopy temperature is 15-30 DEG C, humidity >=70%；The 16-18 days, at 10 points in the morning all deployed to 4 points or so of afternoon, interior external sunshade；19-22 My god, at 10 points in the morning deploys to 4 points or so of afternoon, external sunshade, afterwards full exposure hardening, until semi-lignified, the thickening greening of blade, Can field planting or transplanting.
- 2. the method for Pterospermi Heterophylli tissue culture outside sprout-cultivating-bottle radication according to claim 1, it is characterised in that the tissue-cultured seedling tissue-cultured seedling In rejuvenation, modified MS medium is：NH4NO3500mg/L, KNO31900mg/L, MgSO4·7H2O 370mg/L, KH2PO4 225mg/L, CaCl2·2H2O 440mg/L, MnSO4·4H2O 16.9mg/L, ZnSO4·7H2O 8.6mg/L, H3BO3 6.2mg/L, KI 0.83mg/L, Na2MoO4·2H2O 0.25mg/L, CuSO4·5H2O 0.025mg/L, CoCl2·6H2O 0.025mg/L, FeSO4·7H2O 27.8mg/L, Na2-EDTA·2H2O 37.3mg/L, glycine 2.0mg/L, thiamine hydrochloride Plain 2.0mg/L, puridoxine hydrochloride 0.5mg/L, IV B nicotinic acid 2.0mg/L, inositol 100mg/L.
- 3. the method for Pterospermi Heterophylli tissue culture outside sprout-cultivating-bottle radication according to claim 1, it is characterised in that institute in outside sprout-cultivating-bottle step State the mixture that seedling medium is peat, vermiculite and perlite；Preferably, the weight ratio of the peat, vermiculite and perlite is 3:1:1。
- 4. the method for Pterospermi Heterophylli tissue culture outside sprout-cultivating-bottle radication according to claim 3, it is characterised in that the seedling medium disappears Malicious method is：Peat is spread out to be exposed to the sun under the sun 2-3 days and sterilized, the peat pack after sterilization is standby, will disappear before rooting and transplant Peat, vermiculite and perlite after poison mix in proportion, and dry mixing uniformly obtains mixed-matrix, with 0.3% liquor potassic permanganate pair Mixed-matrix carries out disinfection to mixed-matrix and moistened, and mixed-matrix at least vexed close one day after sterilization produces seedling medium.
- 5. the method for Pterospermi Heterophylli tissue culture outside sprout-cultivating-bottle radication according to claim 1, it is characterised in that institute in outside sprout-cultivating-bottle step It is plastic preserving box with a lid to state container of taking root；Preferably, the specification of the plastic preserving box is 20cm × 13cm × 12cm.
- 6. the method for Pterospermi Heterophylli tissue culture outside sprout-cultivating-bottle radication according to claim 1, it is characterised in that institute in outside sprout-cultivating-bottle step Root-growing agent is stated to be formed by indolebutyric acid and methyl α-naphthyl acetate mixture；Preferably, the mass ratio of the indolebutyric acid and methyl α-naphthyl acetate is 3:2.
- 7. the method for Pterospermi Heterophylli tissue culture outside sprout-cultivating-bottle radication according to claim 1, it is characterised in that institute in outside sprout-cultivating-bottle step It is 800ppm to state root-growing agent.
- 8. the method for Pterospermi Heterophylli tissue culture outside sprout-cultivating-bottle radication according to claim 1, it is characterised in that the tissue-cultured seedling is Fujian half Maple lotus tissue-cultured seedling or leaflet Pterospermi Heterophylli tissue-cultured seedling.
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