CN110122333A - A kind of method that flame tree seed stratification is taken root - Google Patents
A kind of method that flame tree seed stratification is taken root Download PDFInfo
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- CN110122333A CN110122333A CN201910521090.1A CN201910521090A CN110122333A CN 110122333 A CN110122333 A CN 110122333A CN 201910521090 A CN201910521090 A CN 201910521090A CN 110122333 A CN110122333 A CN 110122333A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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Abstract
The invention discloses a kind of methods that flame tree seed stratification is taken root, acquisition, processing and disinfection including flame tree seed, the disinfection of laboratory and experimental article, the preparation of culture medium and the inoculation of seed and culture and etc..Method particularly includes: acquisition current year non-hibernating eggs, it is impregnated in boiling water and clear water, kind of a skin is obtained to be disinfected after the seed of swelling, it is inoculated into the culture medium of 1/2MS+IBA 0.5-2mg/L+NAA 0.05-0.2mg/L and cultivates, culture control intensity of illumination 1200-1500lx, 10-12 hours/day of light application time, temperature (23 ± 2) DEG C.Seed sterilization is aseptic seed by the present invention, and aseptic seed is allowed further to germinate, take root, and relative to traditional seedling raising method, has the advantages that proliferation rate height and low in cost.
Description
Technical field
The present invention relates to plant seed germination technical fields, and in particular to a kind of side that flame tree seed stratification is taken root
Method.
Background technique
Flame tree (scientific name: Delonix regia) is named in " plumage of Ye Rufei phoenix, if the hat of flower phoenix ", alias phoenix
Tree, phoenix flower, fiery phoenix, buttercup, flame wood, the flourishing principal columns of a hall, safflower principal columns of a hall tree (Guangzhou), fiery tree, shadow tree, the foreign principal columns of a hall and fire tree
Deng.Flame tree originates in African Madagascar, is world-renowned Landscape Trees, in torrid areas, introducing and planting is more general
Time.The plant belonged to for pulse family flame tree.Deciduous tree, 20 meters of Gao Keda.Tree crown is broad.Two times pinnate compound leaves, leaflet are long oval
Shape.Summer-flowering, raceme, Hua great is red, glossy.Pod is wooden, long up to 50 centimetres.Flame tree is because scarlet or orange
The flower of color cooperates viridian pinnate compound leaf, is known as one of most bright trees in the world.It with magnificent flower with
Greenery as pteridophyte are outstanding.Grow trees of many tropical and subtropical zone places as decoration and the masking sun in the world.
Flame tree plant is tall and big, sagging due to the horizontal exhibition of tree crown, dense large and catch the wind, and serves as shade tree in torrid areas
Role.Pyrophilous, more days environment of property, must can vegetate in sunny prescription.Be distributed in SOUTHERN CHINA and the west and south,
Each torrid zone place of source area Madagascar and the world.Flame tree is tropical tree species, and plantation starts to bloom for 6~8 years, pyrophilous more
Wet and sunny environment grows 20~30 DEG C of thermophilic, can not resist cold, and winter temperature is not less than 10 DEG C.With it is deep it is fertile, rich in having
The sandy loam of machine matter is advisable；Be afraid of ponding, draining must be good, more drought-resistant；Barren-resistant soil.General 1 year life height reachable 1.5~
It is 2 meters, 2 years raw high up to 3~4 meters, 6~8 years beginnings of plantation flower.
The common propagation method of flame tree is seeding and seedling raising.About 400 grams of thousand grain weigth.The hard water suction of kind skin is difficult, needs
It is impregnated with water.With a program request method sowing, seedling is more sensitive to frost, can apply comprehensive fertilizer in early days, Shaoshi nitrogenous fertilizer, autumn has set in
It should stop applying fertilizer later, promote its lignifying early.It not yet falls off, can manually cut off, and covered with film into winter, such as blade
Lid or single plant wrap up frost prevention.1 year raw seedling can be out of the garden field planting.But percentage of seedgermination is lower, the especially kind without any processing
Sub- germination percentage only has 10% or so, and normal development is only half or so of seedling, and passes through physics and wipe the germination percentage to break in the seed coat
Also only have 31%, and physics wipes the mode that breaks in the seed coat and is difficult to operate, and easily destroys Interior Seed structure, prevent it from germinateing or
It cannot grow up healthy and sound as plant.In addition, the service life of flame tree seed is also very short, saves a Nian Houqi germination percentage and be greatly lowered.
Therefore method for tissue culture is utilized, it is especially that material quickly breed the flame tree nursery stock produced with disease-free with seed seedling
The advantages that malicious, sterile, and breeding coefficient is high.
It is aseptically connect after carrying out a series of processing before exploration discovery sprouts flame tree seed after study
Enter that its germination can be effectively facilitated in specific culture medium to take root and generates seedling.Therefore it is proposed that a kind of content is simple
It is single, low-cost seed treatment and seed germination method.
Summary of the invention
In view of the deficienciess of the prior art, both being wasted present invention aim to address flame tree percentage of seedgermination is low
Seed, and the problem of increase investment, therefore a kind of simple and low-cost seed treatment is provided and is promoted using culture medium
The method that germination takes root.
To achieve the above object, the present invention provides the following technical scheme that
A kind of method that flame tree seed stratification is taken root, comprising the following steps:
Step 1: the acquisition of seed, acquisition current year non-hibernating eggs select full disease-free seed in the same size；
Step 2: testing the disinfection and preparation of preceding laboratory and experiment appliance, and prepare 1/2MS culture medium and hormone mother liquor；
Step 3: the seed that step 1 acquires is soaked seed in 100 DEG C of boiling water, makes its natural cooling, so by the processing of seed
After be put in clear water and impregnate 72h, every 12h changes a water during immersion, obtains kind of skin by the seed of swelling, then carries out conventional disappears
Poison processing: tap water rinses 30min first, then with 70% alcohol disinfecting 8min, with aseptic water washing 3 times, finally with 10% time
Sodium chlorate solution sterilizes 8min, is rushed 5 times on superclean bench with sterile water, impregnates and slightly to be shaken in flushing process
It swings；
Step 4: the inoculation and culture of seed are impregnated before inoculation in the superclean bench Gibberellins solution of 60mg/L
Then 30min is seeded in the 1/2MS culture medium added with agar and hormone and cultivates；
The formula of the 1/2MS culture medium of the addition hormone mother liquor are as follows: 1/2MS+IBA 0.5-2mg/L+NAA0.05-
0.2mg/L。
As the present invention further scheme: the superclean bench needed before use in advance 30min open ultraviolet lamp into
Row sterilizing, when operation, close ultraviolet lamp, open fan and simultaneously place alcolhol burner inside the superclean bench.
As further scheme of the invention: the formula of the 1/2MS culture medium of the addition hormone mother liquor are as follows: 1/2MS+
IBA 0.8-1.2mg/L+NAA0.08-0.12mg/L。
As further scheme of the invention: the formula of the 1/2MS culture medium of the addition hormone mother liquor are as follows: 1/2MS+
IBA 1mg/L+NAA0.1mg/L。
As further scheme of the invention: the 1/2MS culture medium of preparation described in step 2 uses preceding in 121 DEG C of high pressures
Pot sterilizing 15-18min.
As further scheme of the invention: the water-soluble needle tubing filtering head of the hormone mother liquor of preparation described in step 2
It is put in refrigerator and freezes after filtering.
As further scheme of the invention: cooling on superclean bench after the completion of the 1/2MS medium sterilization
To 55 DEG C ± 2 DEG C, it is dispensed into culture bottle after agar and hormone is added, every bottle of culture medium 50mL ± 5mL.
As the present invention further scheme: the inoculation operation of seed described in step 4 is with sterilizing on alcolhol burner
Tweezers clamp seed and lie against and submerge the 1/3-1/2 of seed in culture medium.
As further scheme of the invention: 4-5 seed of every bottle of inoculation of medium described in step 4.
As further scheme of the invention: condition of culture described in step 4 are as follows: intensity of illumination 1200-1500lx,
10-12 hours/day of light application time, 23 DEG C ± 2 DEG C of temperature
Compared with prior art, the invention has the following advantages that
(1) present invention sterilizes to flame tree seed using conventional disinfection method, can both effectively kill and be attached to
The bacterium of the surface of the seed and fungal spores, and kind of a neutron poisoning can be prevented, the vitality of seed is kept, good no strain is obtained
Son.
(2) present invention cultivates seed in the 1/2MS culture medium containing auxin, utilizes auxin
Growth-promoting root and growth promoting function, so that the germination of flame tree seed be promoted to take root, therefore, the present invention is with high security and can
The good advantage of duration.
(3) technological operation is simple, low in cost, reproducible, and can increase substantially flame tree seed germination percentage and
Rooting rate has preferable economic benefit and social benefit.
(4) its germination percentage is greatly improved to seed using kind of a skin processing is impregnated in the present invention, therefore, present invention letter
It is single-phase to operation, and achieve higher benefit.
Specific embodiment
Technical solution in the embodiment of the present invention is understood and is fully described by below, it is clear that described reality
It applies example to be only a part of the embodiment of the present invention, instead of all the embodiments.
Embodiment 1:
The method that flame tree seed stratification is taken root, comprising the following steps:
Step 1: the acquisition of seed, acquisition current year non-hibernating eggs select full disease-free seed in the same size；
Step 2: experiment before give laboratory UV sterilize 30min, with pressure cooker sterilizing culture bottle, tweezers, tap water and
Liquid transfer gun head；It prepares 1/2MS culture medium and required hormone mother liquor, configured culture medium needs to carry out pressure cooker before use
Sterilizing.Prepared hormone, which needs to be put in refrigerator after being filtered with water-soluble needle tubing filtering head, to be freezed.It is wherein high
The use condition for pressing pot is 121 DEG C, 15-18min；
Step 3: the seed that step 1 acquires being soaked seed in 100 DEG C of boiling water, makes its natural cooling, is then put in clear water
Middle immersion 72h, every 12h changes a water during immersion, obtains kind of skin by the seed of swelling.
Conventional explant disinfection is carried out after the completion of impregnating.First the seed soaked is placed under tap water and is rinsed
30min.Tap is opened, seed is placed on the mouth of a river in the following, being directly rinsed, water should not be too large, and crossing conference will impregnate
Seed break through, influence the integrality of seed.It is put into progress soaking flushing 8min in 70% alcohol after the completion of rinsing, impregnated
Cheng Zhongke is slightly swayed, and with the aseptic water washing to have sterilized 3 times after the completion of impregnating, is then soaked again with 10% liquor natrii hypochloritis
Bubble rinses 8min, also needs to be swayed in flushing process.In superclean bench with aseptic water washing five times after the completion of flushing.Pay attention to
Have in superclean bench aseptic water washing after the completion of sodium hypochlorite soaking disinfection, before alcohol soaking flushing can not be super
Net workbench carries out.Wherein superclean bench needed before use in advance 30min open ultraviolet lamp and sterilize, when operation, closes ultraviolet
Lamp opens fan and places alcolhol burner inside the superclean bench.
Step 4: configured culture medium being put into after the completion of sterilizing in autoclave, superclean bench is put it into, to it
When being cooled to 55 DEG C or so, adds agar and be dispensed into culture bottle after measuring hormone addition with liquid-transfering gun, every bottle of culture
Base 50mL or so.Notice that the temperature that culture medium can not be made cooling is too low, because culture medium can solidify when temperature is too low, cannot dispense
And carry out subsequent operation.Note: all of above operation must be completed in superclean bench.
After liquor natrii hypochloritis sterilizes and uses aseptic water washing 5 times, seed is put into the red mould of prepared 60mg/L
30min is impregnated in plain solution, after the completion of immersion in gibberellin, can be inoculated with.It is cooling solidifying to above-mentioned prepared culture medium
Gu being inoculated with after the completion.Seed is gently clamped with tweezers when inoculation to lie against in culture medium and submerge seed 1/3-1/2
It is fixed in culture medium, 4-5 seed is put in every bottle, is placed uniform.Tweezers are used on alcohol when placing seed with tweezers
Fire disinfection.Made a record on culture bottle after the completion of inoculation, culture bottle taken out from superclean bench, be placed on (23 ± 2) DEG C,
Light application time is that 10-12 hours/day, intensity of illumination are cultivated under conditions of being 1200-1500lx.It 4-5 days after inoculation will
The bud point occurred in vain is emerged slightly.
Present invention selection, which can promote the hormone in medium component that flame tree germination takes root, to be had:
A、1/2MS+IBA1mg/L
B、1/2MS+IBA1mg/L+NAA0.1mg/L
C、1/2MS+IBA0.5mg/L+NAA0.2mg/L
D、1/2MS+IBA2mg/L+NAA0.05mg/L
E、1/2MS+IBA1mg/L+2,4-D 0.2mg/L
F、1/2MS+IBA0.8mg/L+2,4-D 0.2mg/L
G、1/2MS+IBA0.8mg/L+2,4-D 0.5mg/L
H、1/2MS+IBA0.8mg/L+2,4-D 1.0mg/L
I、1/2MS+IBA1.3mg/L+2,4-D 0.5mg/L
J、1/2MS+IBA1.3mg/L+2,4-D 1.0mg/L
Wherein culture medium is that the effect of 1/2MS+IBA (indolebutyric acid) 1mg/L+NAA (methyl α-naphthyl acetate) 0.1mg/L is best, hair
Bud rate reaches 47% or so, and rooting rate reaches 33% or so.
1/2MS formula provided by the invention are as follows: a great number of elements: ammonium nitrate NH4NO3825mg/L, potassium nitrate KNO3
950mg/L, calcium chloride dihydrate CaCl2·2H2O 220mg/L, epsom salt MgSO4·7H2O 185mg/L, potassium dihydrogen phosphate
KH2PO485mg/L；
Microelement: potassium iodide KI 0.415mg/L, boric acid H3BO33.1mg/L, four water manganese sulfate MnSO4·4H2O
11.15mg/L, white vitriol ZnSO4·7H2O 4.3mg/L, Sodium Molybdate Dihydrate Na2MoO4·2H2O 0.125mg/L, five water
Copper sulphate CuSO4·5H2O 0.0125mg/L, CoCL2 6H2O CoCl2·6H2O 0.0125mg/L；
Molysite: ferrous sulfate heptahydrate FeSO4·7H2O 27.8mg/L, two water disodium ethylene diamine tetraacetate Na2-EDTA·
2H2O 37.3mg/L；
Organic substance: inositol 100mg/L, niacin VB5 or VPP 0.5mg/L, puridoxine hydrochloride VB6 0.5mg/L, hydrochloric acid
Thiamine VB1 0.1mg/L, glycine 2.0mg/L；
Sucrose 30g/L；
Agar 7g/L.
The design scheme and germination percentage, rooting rate of 1 flame tree seed of table
Note :+, expression has carried out this operation；, indicate not carry out this operation.
The germination percentage and rooting rate of the seed normally handled it can be seen from above-mentioned germination percentage and rooting rate data
Higher than the untreated seed of a certain item, especially 1/2MS culture medium without the seed of sterilization treatment.
Embodiment 2:
(1) acquisition current year non-hibernating eggs, selects full disease-free seed in the same size；
(2) disinfection and preparation in preceding laboratory and experiment appliance are tested, and prepares 1/2MS culture medium and IBA, NAA hormone
Mother liquor, the 1/2MS culture medium of preparation is using preceding in 121 DEG C of pressure cookers sterilizing 15min；IBA, NAA hormone mother liquor water of preparation
It is put in refrigerator and freezes after the needle tubing filtering head filtering of dissolubility；
(3) seed of acquisition is soaked seed in 100 DEG C of boiling water, makes its natural cooling, is then put in clear water and impregnates
72h, every 12h changes a water during immersion.Kind of skin is obtained by the seed of swelling, then carries out routine disinfection processing: first originally
Water rinses 30min, then is finally soaked with 10% liquor natrii hypochloritis with 70% alcohol soaking flushing 8min with aseptic water washing 3 times
Bubble rinses 8min, uses aseptic water washing 5 times on superclean bench, slightly to shake during soaking flushing；
(4) 30min is impregnated in the superclean bench Gibberellins solution of 60mg/L before inoculation, being then seeded in formula is 1/
It is cultivated in the culture medium of 2MS+IBA 0.5mg/L+NAA 0.2mg/L, inoculation operation is with the tweezers sterilized on alcolhol burner
Seed is gently clamped to lie against in culture medium and seed 1/3-1/2 is made to submerge fixation in culture medium.Condition of culture are as follows: illumination is strong
Spend 1200lx, 12 hour/day of light application time, temperature (23 ± 2) DEG C.
Note: wherein superclean bench needed before use in advance 30min open ultraviolet lamp and sterilize, when operation, closes ultraviolet
Lamp opens fan and places alcolhol burner inside the superclean bench.
Embodiment 3:
(1) acquisition current year non-hibernating eggs, selects full disease-free seed in the same size；
(2) disinfection and preparation in preceding laboratory and experiment appliance are tested, and prepares 1/2MS culture medium and IBA, NAA hormone
Mother liquor, the 1/2MS culture medium of preparation is using preceding in 121 DEG C of pressure cookers sterilizing 18min；IBA, NAA hormone mother liquor water of preparation
It is put in refrigerator and freezes after the needle tubing filtering head filtering of dissolubility；
(3) seed of acquisition is soaked seed in 100 DEG C of boiling water, makes its natural cooling, is then put in clear water and impregnates
72h, every 12h changes a water during immersion.Kind of skin is obtained by the seed of swelling, then carries out routine disinfection processing: first originally
Water rinses 30min, then is finally soaked with 10% liquor natrii hypochloritis with 70% alcohol soaking flushing 8min with aseptic water washing 3 times
Bubble rinses 8min, uses aseptic water washing 5 times on superclean bench, slightly to shake during soaking flushing；
(4) 30min is impregnated in the superclean bench Gibberellins solution of 60mg/L before inoculation, being then seeded in formula is 1/
It is cultivated in the culture medium of 2MS+IBA 2mg/L+NAA 0.05mg/L, inoculation operation is light with the tweezers sterilized on alcolhol burner
Seed is gently clamped to lie against in culture medium and seed 1/3-1/2 is made to submerge fixation in culture medium.Condition of culture are as follows: intensity of illumination
1500lx, 10 hour/day of light application time, temperature (23 ± 2) DEG C.
Note: wherein superclean bench needed before use in advance 30min open ultraviolet lamp and sterilize, when operation, closes ultraviolet
Lamp opens fan and places alcolhol burner inside the superclean bench.
Embodiment 4:
(1) acquisition current year non-hibernating eggs, selects full disease-free seed in the same size；
(2) disinfection and preparation in preceding laboratory and experiment appliance are tested, and prepares 1/2MS culture medium and IBA, NAA hormone
Mother liquor, the 1/2MS culture medium of preparation is using preceding in 121 DEG C of pressure cookers sterilizing 17min；IBA, NAA hormone mother liquor water of preparation
It is put in refrigerator and freezes after the needle tubing filtering head filtering of dissolubility；
(3) seed of acquisition is soaked seed in 100 DEG C of boiling water, makes its natural cooling, is then put in clear water and impregnates
72h, every 12h changes a water during immersion.Kind of skin is obtained by the seed of swelling, then carries out routine disinfection processing: first originally
Water rinses 30min, then is finally soaked with 10% liquor natrii hypochloritis with 70% alcohol soaking flushing 8min with aseptic water washing 3 times
Bubble rinses 8min, uses aseptic water washing 5 times on superclean bench, slightly to shake during soaking flushing；
(4) 30min is impregnated in the superclean bench Gibberellins solution of 60mg/L before inoculation, being then seeded in formula is 1/
It is cultivated in the culture medium of 2MS+IBA 1.2mg/L+NAA 0.13mg/L, inoculation operation is with the tweezers sterilized on alcolhol burner
Seed is gently clamped to lie against in culture medium and seed 1/3-1/2 is made to submerge fixation in culture medium.Condition of culture are as follows: illumination is strong
Spend 1400lx, 11 hour/day of light application time, temperature (23 ± 2) DEG C.
Note: wherein superclean bench needed before use in advance 30min open ultraviolet lamp and sterilize, when operation, closes ultraviolet
Lamp opens fan and places alcolhol burner inside the superclean bench.
Embodiment 5:
(1) acquisition current year non-hibernating eggs, selects full disease-free seed in the same size；
(2) disinfection and preparation in preceding laboratory and experiment appliance are tested, and prepares 1/2MS culture medium and IBA, NAA hormone
Mother liquor, the 1/2MS culture medium of preparation is using preceding in 121 DEG C of pressure cookers sterilizing 16min；IBA, NAA hormone mother liquor water of preparation
It is put in refrigerator and freezes after the needle tubing filtering head filtering of dissolubility；
(3) seed of acquisition is soaked seed in 100 DEG C of boiling water, makes its natural cooling, is then put in clear water and impregnates
72h, every 12h changes a water during immersion.Kind of skin is obtained by the seed of swelling, then carries out routine disinfection processing: first originally
Water rinses 30min, then is finally soaked with 10% liquor natrii hypochloritis with 70% alcohol soaking flushing 8min with aseptic water washing 3 times
Bubble rinses 8min, uses aseptic water washing 5 times on superclean bench, slightly to shake during soaking flushing；
(4) 30min is impregnated in the superclean bench Gibberellins solution of 60mg/L before inoculation, being then seeded in formula is 1/
It is cultivated in the culture medium of 2MS+IBA 1.7mg/L+NAA 0.09mg/L, inoculation operation is with the tweezers sterilized on alcolhol burner
Seed is gently clamped to lie against in culture medium and seed 1/3-1/2 is made to submerge fixation in culture medium.Condition of culture are as follows: illumination is strong
Spend 1400lx, 11 hour/day of light application time, temperature (23 ± 2) DEG C.
Note: wherein superclean bench needed before use in advance 30min open ultraviolet lamp and sterilize, when operation, closes ultraviolet
Lamp opens fan and places alcolhol burner inside the superclean bench.
Embodiment 6:
(1) acquisition current year non-hibernating eggs, selects full disease-free seed in the same size；
(2) disinfection and preparation in preceding laboratory and experiment appliance are tested, and prepares 1/2MS culture medium and IBA, NAA hormone
Mother liquor, the 1/2MS culture medium of preparation is using preceding in 121 DEG C of pressure cookers sterilizing 18min；IBA, NAA hormone mother liquor water of preparation
It is put in refrigerator and freezes after the needle tubing filtering head filtering of dissolubility；
(3) seed of acquisition is soaked seed in 100 DEG C of boiling water, makes its natural cooling, is then put in clear water and impregnates
72h, every 12h changes a water during immersion.Kind of skin is obtained by the seed of swelling, then carries out routine disinfection processing: first originally
Water rinses 30min, then is finally soaked with 10% liquor natrii hypochloritis with 70% alcohol soaking flushing 8min with aseptic water washing 3 times
Bubble rinses 8min, uses aseptic water washing 5 times on superclean bench, slightly to shake during soaking flushing；
(4) 30min is impregnated in the superclean bench Gibberellins solution of 60mg/L before inoculation, being then seeded in formula is 1/
It is cultivated in the culture medium of 2MS+IBA 0.8mg/L+NAA 0.08mg/L, inoculation operation is with the tweezers sterilized on alcolhol burner
Seed is gently clamped to lie against in culture medium and seed 1/3-1/2 is made to submerge fixation in culture medium.Condition of culture are as follows: illumination is strong
Spend 1400lx, 11 hour/day of light application time, temperature (23 ± 2) DEG C.
Note: wherein superclean bench needed before use in advance 30min open ultraviolet lamp and sterilize, when operation, closes ultraviolet
Lamp opens fan and places alcolhol burner inside the superclean bench.
Embodiment 7:
(1) acquisition current year non-hibernating eggs, selects full disease-free seed in the same size；
(2) disinfection and preparation in preceding laboratory and experiment appliance are tested, and prepares 1/2MS culture medium and IBA, NAA hormone
Mother liquor, the 1/2MS culture medium of preparation is using preceding in 121 DEG C of pressure cookers sterilizing 15min；IBA, NAA hormone mother liquor water of preparation
It is put in refrigerator and freezes after the needle tubing filtering head filtering of dissolubility；
(3) seed of acquisition is soaked seed in 100 DEG C of boiling water, makes its natural cooling, is then put in clear water and impregnates
72h, every 12h changes a water during immersion.Kind of skin is obtained by the seed of swelling, then carries out routine disinfection processing: first originally
Water rinses 30min, then is finally soaked with 10% liquor natrii hypochloritis with 70% alcohol soaking flushing 8min with aseptic water washing 3 times
Bubble rinses 8min, uses aseptic water washing 5 times on superclean bench, slightly to shake during soaking flushing；
(4) 30min is impregnated in the superclean bench Gibberellins solution of 60mg/L before inoculation, being then seeded in formula is 1/
It is cultivated in the culture medium of 2MS+IBA 1.2mg/L+NAA 0.12mg/L, inoculation operation is with the tweezers sterilized on alcolhol burner
Seed is gently clamped to lie against in culture medium and seed 1/3-1/2 is made to submerge fixation in culture medium.Condition of culture are as follows: illumination is strong
Spend 1400lx, 11 hour/day of light application time, temperature (23 ± 2) DEG C.
Note: wherein superclean bench needed before use in advance 30min open ultraviolet lamp and sterilize, when operation, closes ultraviolet
Lamp opens fan and places alcolhol burner inside the superclean bench.
Embodiment 8:
(1) acquisition current year non-hibernating eggs, selects full disease-free seed in the same size；
(2) disinfection and preparation in preceding laboratory and experiment appliance are tested, and prepares 1/2MS culture medium and IBA, NAA hormone
Mother liquor, the 1/2MS culture medium of preparation is using preceding in 121 DEG C of pressure cookers sterilizing 17min；IBA, NAA hormone mother liquor water of preparation
It is put in refrigerator and freezes after the needle tubing filtering head filtering of dissolubility；
(3) seed of acquisition is soaked seed in 100 DEG C of boiling water, makes its natural cooling, is then put in clear water and impregnates
72h, every 12h changes a water during immersion.Kind of skin is obtained by the seed of swelling, then carries out routine disinfection processing: first originally
Water rinses 30min, then is finally soaked with 10% liquor natrii hypochloritis with 70% alcohol soaking flushing 8min with aseptic water washing 3 times
Bubble rinses 8min, uses aseptic water washing 5 times on superclean bench, slightly to shake during soaking flushing；
(4) 30min is impregnated in the superclean bench Gibberellins solution of 60mg/L before inoculation, being then seeded in formula is 1/
2MS+IBA1.1mg/L+NAA0.08mg/L culture medium in cultivate, inoculation operation be light with the tweezers sterilized on alcolhol burner
Seed is gently clamped to lie against in culture medium and seed 1/3-1/2 is made to submerge fixation in culture medium.Condition of culture are as follows: intensity of illumination
1400lx, 11 hour/day of light application time, temperature (23 ± 2) DEG C.
Note: wherein superclean bench needed before use in advance 30min open ultraviolet lamp and sterilize, when operation, closes ultraviolet
Lamp opens fan and places alcolhol burner inside the superclean bench.
Embodiment 9:
(1) acquisition current year non-hibernating eggs, selects full disease-free seed in the same size；
(2) disinfection and preparation in preceding laboratory and experiment appliance are tested, and prepares 1/2MS culture medium and IBA, NAA hormone
Mother liquor, the 1/2MS culture medium of preparation is using preceding in 121 DEG C of pressure cookers sterilizing 16min；IBA, NAA hormone mother liquor water of preparation
It is put in refrigerator and freezes after the needle tubing filtering head filtering of dissolubility；
(3) seed of acquisition is soaked seed in 100 DEG C of boiling water, makes its natural cooling, is then put in clear water and impregnates
72h, every 12h changes a water during immersion.Kind of skin is obtained by the seed of swelling, then carries out routine disinfection processing: first originally
Water rinses 30min, then is finally soaked with 10% liquor natrii hypochloritis with 70% alcohol soaking flushing 8min with aseptic water washing 3 times
Bubble rinses 8min, uses aseptic water washing 5 times on superclean bench, slightly to shake during soaking flushing；
(4) 30min is impregnated in the superclean bench Gibberellins solution of 60mg/L before inoculation, being then seeded in formula is 1/
2MS+IBA1mg/L+NAA0.1mg/L culture medium in cultivate, inoculation operation be with the tweezers sterilized on alcolhol burner gently
Seed is clamped to lie against in culture medium and seed 1/3-1/2 is made to submerge fixation in culture medium.Condition of culture are as follows: intensity of illumination
1400lx, 11 hour/day of light application time, temperature (23 ± 2) DEG C.
Note: wherein superclean bench needed before use in advance 30min open ultraviolet lamp and sterilize, when operation, closes ultraviolet
Lamp opens fan and places alcolhol burner inside the superclean bench.
Comparative example:
(1) acquisition current year non-hibernating eggs, selects full disease-free seed in the same size；
(2) sufficient amount of primary loam is acquired from flame tree location, is mixed with fine sand according to the ratio of 1:1
And it is divided in flowerpot；
(3) guarantee the soil environment needed for seed is sprouted, to its suitable temperature, humidity and required nutrient.
Using the method for above-described embodiment 2-9 and comparative example, flame tree seed is cultivated, it can be with from the result of following table
Find out, the technical indicator of embodiment is far more than comparative example.
2 embodiment design scheme of table is compared with comparative example effect
| Embodiment | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | Comparative example |
| Germination percentage % | 47.5 | 43.2 | 42.2 | 38.5 | 37.1 | 39.2 | 42.1 | 40.2 | 17.2 |
| Rooting rate % | 35.7 | 37.2 | 35.4 | 32.2 | 34.5 | 31.7 | 37.5 | 34.4 | 14.8 |
The foregoing is only a preferred embodiment of the present invention, but scope of protection of the present invention is not limited thereto,
Anyone skilled in the art in the technical scope disclosed by the present invention, according to the technique and scheme of the present invention and its
Inventive concept is subject to equivalent substitution or change, should be covered by the protection scope of the present invention.
Claims (10)
1. a kind of method that flame tree seed stratification is taken root, which comprises the following steps:
Step 1: the acquisition of seed, acquisition current year non-hibernating eggs select full disease-free seed in the same size；
Step 2: testing the disinfection and preparation of preceding laboratory and experiment appliance, and prepare 1/2MS culture medium and hormone mother liquor；
Step 3: the seed that step 1 acquires is soaked seed in 100 DEG C of boiling water, makes its natural cooling, then put by the processing of seed
72h is impregnated in clear water, every 12h changes a water during immersion, obtains kind of skin by the seed of swelling, then carries out at routine disinfection
Reason: tap water rinses 30min first, then with 70% alcohol disinfecting 8min, with aseptic water washing 3 times, finally with 10% hypochlorous acid
Sodium solution sterilizes 8min, is rushed 5 times on superclean bench with sterile water, impregnates and slightly to be shaken in flushing process；
Step 4: the inoculation and culture of seed impregnate 30min in the superclean bench Gibberellins solution of 60mg/L before inoculation, so
It is seeded in the 1/2MS culture medium added with agar and hormone and cultivates afterwards；
The formula of the 1/2MS culture medium of the addition hormone mother liquor are as follows: 1/2MS+IBA0.5-2mg/L+NAA0.05-0.2mg/L.
2. the method that flame tree seed stratification according to claim 1 is taken root, which is characterized in that the ultra-clean work
Platform needed before use in advance 30min open ultraviolet lamp and sterilize, when operation, closes ultraviolet lamp, opens fan and in the ultra-clean work
Make to place alcolhol burner inside platform.
3. the method that flame tree seed stratification according to claim 1 is taken root, which is characterized in that the addition hormone
The formula of the 1/2MS culture medium of mother liquor are as follows: 1/2MS+IBA0.8-1.2mg/L+NAA0.08-0.12mg/L.
4. the method that flame tree seed stratification according to claim 1 is taken root, which is characterized in that the addition hormone
The formula of the 1/2MS culture medium of mother liquor are as follows: 1/2MS+IBA1mg/L+NAA0.1mg/L.
5. the method that flame tree seed stratification according to claim 1 is taken root, which is characterized in that match described in step 2
The 1/2MS culture medium of system is using preceding in 121 DEG C of pressure cookers sterilizing 15-18min.
6. the method that flame tree seed stratification according to claim 1 is taken root, which is characterized in that match described in step 2
The hormone mother liquor of system is put in refrigerator after being filtered with water-soluble needle tubing filtering head and freezes.
7. the method that flame tree seed stratification according to claim 5 is taken root, which is characterized in that the 1/2MS training
After the completion of supporting base sterilizing, it is cooled to 55 DEG C ± 2 DEG C on superclean bench, is dispensed into culture bottle after agar and hormone is added,
Every bottle of culture medium 50mL ± 5mL.
8. the method that flame tree seed stratification according to claim 1 is taken root, which is characterized in that planted described in step 4
The inoculation operation of son is to be clamped seed with the tweezers sterilized on alcolhol burner and lain against in culture medium and to make the 1/3- of seed
1/2 submerges in culture medium.
9. the method that flame tree seed stratification according to claim 1 is taken root, which is characterized in that every described in step 4
Bottle 4-5 seed of inoculation of medium.
10. the method that flame tree seed stratification according to claim 1 is taken root, which is characterized in that described in step 4
Condition of culture are as follows: intensity of illumination 1200-1500lx, 10-12 hours/day of light application time, 23 DEG C ± 2 DEG C of temperature.
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