CN105660398B - A kind of tissue cultivation rapid breeding method of the big wood paint in mao of dam - Google Patents
A kind of tissue cultivation rapid breeding method of the big wood paint in mao of dam Download PDFInfo
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- CN105660398B CN105660398B CN201610023590.9A CN201610023590A CN105660398B CN 105660398 B CN105660398 B CN 105660398B CN 201610023590 A CN201610023590 A CN 201610023590A CN 105660398 B CN105660398 B CN 105660398B
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- 239000003973 paint Substances 0.000 title claims abstract description 39
- 239000002023 wood Substances 0.000 title claims abstract description 34
- 238000009395 breeding Methods 0.000 title claims abstract description 25
- 230000004069 differentiation Effects 0.000 claims abstract description 12
- 239000000463 material Substances 0.000 claims abstract description 9
- 230000001954 sterilising effect Effects 0.000 claims abstract description 9
- 239000000835 fiber Substances 0.000 claims abstract description 6
- 238000004659 sterilization and disinfection Methods 0.000 claims abstract description 6
- 238000003306 harvesting Methods 0.000 claims abstract description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 17
- 239000001963 growth medium Substances 0.000 claims description 15
- 238000000034 method Methods 0.000 claims description 14
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- 229920001817 Agar Polymers 0.000 claims description 9
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- 230000001939 inductive effect Effects 0.000 claims description 8
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- 229930006000 Sucrose Natural products 0.000 claims description 7
- 239000005720 sucrose Substances 0.000 claims description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 6
- 238000005406 washing Methods 0.000 claims description 6
- 206010020649 Hyperkeratosis Diseases 0.000 claims description 5
- 229930182555 Penicillin Natural products 0.000 claims description 4
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 claims description 4
- 239000007788 liquid Substances 0.000 claims description 4
- 239000011159 matrix material Substances 0.000 claims description 4
- 229940049954 penicillin Drugs 0.000 claims description 4
- 230000000249 desinfective effect Effects 0.000 claims description 3
- 239000003344 environmental pollutant Substances 0.000 claims description 3
- 239000002609 medium Substances 0.000 claims description 3
- 229960002523 mercuric chloride Drugs 0.000 claims description 3
- LWJROJCJINYWOX-UHFFFAOYSA-L mercury dichloride Chemical compound Cl[Hg]Cl LWJROJCJINYWOX-UHFFFAOYSA-L 0.000 claims description 3
- 239000003415 peat Substances 0.000 claims description 3
- 235000019362 perlite Nutrition 0.000 claims description 3
- 239000010451 perlite Substances 0.000 claims description 3
- 231100000719 pollutant Toxicity 0.000 claims description 3
- 239000012882 rooting medium Substances 0.000 claims description 3
- 239000002689 soil Substances 0.000 claims description 3
- RTKIYNMVFMVABJ-UHFFFAOYSA-L thimerosal Chemical compound [Na+].CC[Hg]SC1=CC=CC=C1C([O-])=O RTKIYNMVFMVABJ-UHFFFAOYSA-L 0.000 claims description 3
- 229940033663 thimerosal Drugs 0.000 claims description 3
- 239000003610 charcoal Substances 0.000 claims 1
- 230000000694 effects Effects 0.000 claims 1
- 239000007943 implant Substances 0.000 claims 1
- 230000001488 breeding effect Effects 0.000 abstract description 17
- 230000006698 induction Effects 0.000 abstract description 7
- 238000005516 engineering process Methods 0.000 abstract description 6
- 230000004083 survival effect Effects 0.000 abstract description 6
- 238000002844 melting Methods 0.000 abstract description 5
- 230000008018 melting Effects 0.000 abstract description 5
- 239000000203 mixture Substances 0.000 abstract description 4
- 230000002062 proliferating effect Effects 0.000 abstract description 4
- 230000001902 propagating effect Effects 0.000 abstract description 4
- 239000000758 substrate Substances 0.000 abstract description 4
- -1 by sterilization Substances 0.000 abstract 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 14
- 241000196324 Embryophyta Species 0.000 description 14
- 244000044283 Toxicodendron succedaneum Species 0.000 description 10
- 239000004922 lacquer Substances 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- DXRKLUVKXMAMOV-UHFFFAOYSA-N 3-heptadecylcatechol Chemical compound CCCCCCCCCCCCCCCCCC1=CC=CC(O)=C1O DXRKLUVKXMAMOV-UHFFFAOYSA-N 0.000 description 2
- 229930192334 Auxin Natural products 0.000 description 2
- 240000000111 Saccharum officinarum Species 0.000 description 2
- 235000007201 Saccharum officinarum Nutrition 0.000 description 2
- 239000002363 auxin Substances 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 description 2
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- 230000010154 cross-pollination Effects 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 230000035876 healing Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
Abstract
The present invention relates to the tissue cultivation rapid breeding method of the big wood paint in a kind of mao of dam.The fresh axillary bud and terminal bud that the present invention chooses the big wood paint in 5 years excellent hair dams of life are used as explant culture materials, by sterilization, Fiber differentiation, Multiplying culture, culture of rootage, hardening and transplanting and other steps, quickly bred by group culturation rapid propagating technology wood paint big to hair dam, calculated with the big wood paint explant in every 10 hair dams, breeding coefficient is 6.5, proliferating cycle is to calculate for 30 days, except the consume that depollutes, its year breeding amount reaches more than 2,750,000, the problem of big wood in mao dam paints rare numbers is efficiently solved, the demand that large area is quickly bred is realized.And the time and the age of tree of the invention by accurately selecting harvesting explant, the induction differentiation rate of adventitious bud is improved, its differentiation rate is reached more than 85%.In addition, the melting brown rate of the final explant of the present invention is less than 8%, pollution rate is less than 2%, by the way of lid hardening is closed and rational substrate composition substantially increases the survival rate of seedling, survival rate is maintained at more than 90%.
Description
Technical field
The present invention relates to the method for Plant Tissue Breeding in agricultural biotechnologies, the group of the big wood paint in more particularly to a kind of mao dam
Knit culture quick-breeding method.
Background technology
Raw lacquer is that the water in oil emulsion white liquid flowed out is cut out of lacquer tree bast zone, is a kind of physiologic secretion of lacquer tree
Thing.Raw lacquer is China's special product renewable resource, is also time-honored traditional native article, yield account for the 80% of world's total amount with
On.Hubei is that China production Qi great is saved, and hair dam paint is formed under specific weather and altitude conditions, 1954 last century, Zhou Enlai
Premier awards board " the dam paint name hat whole world ", and hair dam paint becomes the Traditional Brand for being famous in the whole nation or even the whole world for this.Hair dam raw lacquer with
Paint matter is dense, and bright in color, fragrant, water content is few, and adhesive force is strong, and dry property is good, and laccol rate is up to more than 80% and made a name
At home and abroad, product finds a good sale in tens countries and regions such as Japan, France, the U.S., Germany, Britain, Hong Kong.And the big wood paint in hair dam is
Wild lacquer tree kind in production, is generally cultivated by the way of root nursery using seed seedling-raising and burying.Due to Rhus cross-pollination
Economic tree, larger differentiation can be produced with seminal propagation, the hereditary capacity of its improved seeds is difficult to keep；Educated using root is buried
Seedling, in addition to only a few is there may be bud mutation, the characteristics of being difficult to degenerate with stabilization, and planting percent is low, reproduction speed is slow, to lacquer tree
Improved seeds and fine individual plant are restricted.And group culturation rapid propagating technology, lacquer tree improved seeds and individual plant heredity can be ensured very well
Stability, and large-scale breeding requirement can be met, group culturation rapid propagating technology is related in the prior art applied to lacquer tree breeding side
The report in face, for example, the patent of Application No. 201410311305.4 discloses a kind of efficient quick propagation method of small wood paint, bag
Selection and sterilization (2) Primary culture (3) Multiplying culture (4) culture of rootage (5) hardening and transplant step of (1) explant are included, is adopted
With the method for carrying out tissue cultures to excellent small wood paint fine individual plant, its breeding coefficient can be made to reach 4~5 times, every 10 effectively
Small wood paint axillary bud, is 4.5 by breeding coefficient, and proliferating cycle is to calculate for 45 days, removes pollution wastage, breeding amount is still up within its year
More than 1750000, efficiently solve the problem of small wood paint is quick to breed so that choiceness material can be widely applied,
But the invention is longer proliferating cycle, and breeding coefficient is relatively low, and the small wood paint of culture easily pollutes, browning, and the proportion of goods damageds are high, are not suitable for hair
The tissue-culturing quick-propagation of the big wood paint in dam.In addition, the patent application of Application No. 201410540495.7 discloses a kind of paint
Set the rapid propagation method of regeneration plant, including it is the sterilization of lacquer tree blade, the induction of callus, the breaking up of callus, raw
The steps such as root induction, acclimatization and transplantses, but the invention avoids browning from lacquer tree blade is explant, can cause induction point
Rate is not high.
The content of the invention
It is an object of the invention to the shortcoming for overcoming prior art, there is provided the tissue culture quick breeding side of the big wood paint in a kind of mao of dam
Method, the inventive method has broken the pattern of the big wooden wild breeding of lacquer tree kind in hair dam, solves the big wood paint survival rate in mao dam, breeding potential
Low, rare numbers technical problems, realize the demand that the big wood paint large area in mao dam is quickly bred.
In order to realize the purpose of the present invention, the present invention is adopted the following technical scheme that：
The tissue cultivation rapid breeding method of the big wood paint in a kind of mao of dam, methods described comprises the following steps：
(1) materials of explant：The fresh axillary bud and terminal bud of the big wood paint in 5 years excellent hair dams of life are plucked in annual the first tenday period of a month in May
It is used as culture materials；
(2) disinfecting process：The branch of harvesting is removed into blade, plant surface is cleaned with water with sterilizing banister brush, pollutant is removed,
2h is rinsed with flowing water, then with after aseptic water washing 3~5 times, cuts into the segment with axillary-bud or top-bud and is positioned over sterile culture
In ware, be quickly moved in superclean bench and carry out disinfection, aseptically with 75% ethanol soak 25s after, be put into containing
In 0.1% mercuric chloride thimerosal of 0.01~0.015% Tween-80,10min is gently agitated for, then with aseptic water washing 5~8 times
Afterwards, explant is placed on sterilized filter paper standby after suck dry moisture；
(3) Fiber differentiation：The wound part of axillary bud and terminal bud contact sterilizing liquid is cut off into 3~4mm, it is perpendicular to insert in Fiber differentiation
On base, temperature is placed in carry out light culture under the conditions of 24 ± 2 DEG C, culture is transferred in the inducing culture newly prepared for 1~2 day again,
Intensity of illumination is positioned over after 3 times repeatedly for 1800lx, light application time is illumination cultivation 20~25 under conditions of 11~12 hours/day
My god, wait to grow 3~4cm sproutings；
(4) Multiplying culture：Sprouting is cut from explant, its Bock Altitude is 2~3cm, 2~3 blades, is erected
Insert on proliferated culture medium, it is 24 ± 2 DEG C to be placed in temperature, intensity of illumination is 1800lx, light application time is the condition of 10 hours/day
Lower culture 25~30 days, when there is 1~2cm of diameter callus in axillary bud base portion and bears 5~8 3~4cm adventitious bud,
Adventitious bud is cut again, transferred in the proliferated culture medium that same recipe is placed under the same conditions, you can make Mao Ba great
Wood paint carries out fast-propagation, and its proliferation times is 6~8 times；
(5) culture of rootage：Propagation seedling high 2~3cm is erected and inserted on root media, it is 24 ± 2 DEG C, light to be placed in temperature
It is 1800lx according to intensity, light application time treats that plant height reaches 3~4cm to be cultivated 25~30 days under conditions of 11~12 hours/day,
1~2cm of root length, more than 4 number/strains of taking root, that is, obtain the rooted seedling of the big wood paint in mao dam, to wait to transplant；
(6) hardening is with transplanting：The bottle seedling that will take root is positioned under the conditions of natural lighting, is carried out in the warm canopy of 24 ± 2 DEG C of room temperature
Lid hardening is closed, rooted seedling is taken out after 7 days, the culture medium of root is cleaned up, is soaked with 80% Bravo after root 5min, is transplanted extremely
Substrate composition is peat soil：Perlite=3：In 2 plastic basket, sufficient root water is drenched, subenvironment relative humidity more than 80% is kept,
It can gradually reduce relative humidity after 7 days, keep matrix moistening, control shading rate is 60%~70%, 26 ± 2 DEG C of temperature, 15~
The seedling survived is transplanted to discrete phase with, a diameter of 10cm of rim of a cup degradable non-woven paper Seedling bag after 20 days, maintenance 2~
3 months, height of seedling moved to nursery lot up to 30cm or so and cultivates seedling.
The formula of inducing culture described in above-mentioned steps (3) is：B5+1.0mg/L 6-BA+0.1mg/L NAA+
30.0g/L sucrose+5.0g/L activated carbon+7.0g/L agar+0.5g/L penicillin, pH value is 5.8~6.0.
Above-mentioned steps (4) described proliferation culture medium formula is：MS+1.0mg/L 6-BA+0.5mg/L NAA+30.0g/L sugarcanes
Sugar+5.0g/L activated carbon+7.0g/L agar, pH is 5.8~6.0, wherein, NH in the proliferation culture medium formula MS4NO3Concentration
For the 1/2 of standard medium.
Above-mentioned steps (5) described prescription of rooting medium is：1/2MS+0.5mg/L IBA+0.5mg/L NAA+30g/L sugarcanes
Sugar+5.0g/L activated carbon+7.0g/L agar, pH is 5.8~6.0.
Compared with prior art, the present invention has following beneficial effect：
(1) present invention is quickly bred by group culturation rapid propagating technology wood paint big to hair dam, can ensure that lacquer tree is excellent well
Non-defective unit kind and individual plant genetic stability, and large-scale breeding demand can be met in a short time, break the wild breeding of the seeds
Pattern reach the purpose of domestication, accomplish scale production, meet the demand in market；
(2) time and the age of tree of the invention by accurately selecting harvesting explant, the induction differentiation rate of adventitious bud is improved,
Its differentiation rate is set to reach more than 85%；
(3) present invention selects low NH during Fiber differentiation4 +, high NO3 -The B5 medium of concentration, and by culture just
Phase is inoculated with repeatedly, add appropriate activated carbon and penicillin and combine the mode of light culture come substantially reduce plant melting brown rate and
Pollution rate, finally makes the melting brown rate of explant be less than 8%, pollution rate is less than 2%, and inductivity reaches more than 85%, achieves noticeable achievement；
(4) present invention using the higher MS culture mediums of nutrient and reduces NH in formula in breeding4 +Content, that is, protect
The nutrient for demonstrate,proving Multiplying culture also reduces melting brown rate simultaneously, and selection basic element of cell division 6-BA is combined with auxin NAA
Formula, makes healing tissue development good, the growth coefficient of Multiple Buds reaches 6~8 times；
(5) present invention is in culture of rootage, and the mode culture that two kinds of auxin (IBA, NAA) of selection are combined promotes
The growth of root is accelerated in the induction differentiation of adventitious root again, its rooting rate is reached more than 99%, 4~5 roots of average every plant of band；
(6) present invention hardening with transplant during, by the way of lid hardening is closed and rational substrate composition is carried significantly
The high survival rate of seedling, makes survival rate be maintained at more than 90%；
(7) present invention can effectively solve the problem that xylophyta the problems such as browning, pollution during tissue culture, with guarantee plant
Excellent inhereditary feature, incubation is safe and simple, and breeding coefficient is high, and the advantages of economical and practical easy maintenance.With every 10
The big wood paint explant in hair dam is calculated, and breeding coefficient is 6.5, and proliferating cycle is to calculate for 30 days, except the consume that depollutes, its year breeding amount
More than 2,750,000 are reached, the problem of big wood in mao dam paints rare numbers is efficiently solved, realizes the demand that large area is quickly bred.
Embodiment
Form is described in further detail again to the above of the present invention by the following examples, but should not manage this
The scope solved as above-mentioned theme of the invention is only limitted to following embodiment, and all technologies realized based on the above of the present invention are equal
Belong to the scope of the present invention.
Embodiment 1
The implementation time of the present embodiment is 2014, and the tissue-culturing rapid propagation of the big wood paint in hair dam is in Hubei Province's forestry scientific research group
Train in laboratory and warmhouse booth and carry out.
The tissue cultivation rapid breeding method of the big wood paint in a kind of mao of dam of the present embodiment, methods described comprises the following steps：
(1) materials of explant：The first tenday period of a month in May, 2015 pluck the fresh axillary bud and terminal bud of the big wood paint in 5 years excellent hair dams of life
It is used as culture materials；
(2) disinfecting process：The branch of harvesting is removed into blade, plant surface is cleaned with water with sterilizing banister brush, pollutant is removed,
2h is rinsed with flowing water, then with after aseptic water washing 3~5 times, cuts into the segment with axillary-bud or top-bud and is positioned over sterile culture
In ware, it is quickly moved in superclean bench and carries out disinfection, aseptically soaks 25s with 75% ethanol, then use sterilized water
After rinsing 3~5 times, it is put into the 0.1% mercuric chloride thimerosal containing 0.01~0.015% Tween-80, is gently agitated for 10min,
Again with after aseptic water washing 5~8 times, explant is placed on sterilized filter paper standby after suck dry moisture；
(3) Fiber differentiation：The formula of inducing culture is B5+1.0mg/L 6-BA+0.1mg/L NAA+30.0g/L sucrose
+ 5.0g/L activated carbon+7.0g/L agar+0.5g/L penicillin, pH value is 5.8~6.0；By axillary bud and terminal bud contact sterilizing liquid
Wound part cuts off 3~4mm, perpendicular to insert on inducing culture, is placed in temperature to carry out light culture, culture 1 under the conditions of 24 ± 2 DEG C
It is transferred to again in the inducing culture newly prepared within~2 days, is positioned over intensity of illumination after 3 times repeatedly for 1800lx, light application time is 11
Continuous light culture 20~25 days under conditions of~12 hours/day, wait to grow 3~4cm sproutings, final through counting this method
The melting brown rate of explant is set to be less than 8%, pollution rate is less than 2%, and inductivity reaches more than 85%.
(4) Multiplying culture：Proliferation culture medium formula is：MS (wherein NH4NO3Deal halves)+1.0mg/L 6-BA+
0.5mg/L NAA+30.0g/L sucrose+5.0g/L activated carbon+7.0g/L agar, pH is 5.8~6.0；In early June, 2014, will
Multiple Buds are cut from explant, and its Bock Altitude is 2~3cm, 2~3 blades, is erected and inserted on proliferated culture medium, put
It it is 24 ± 2 DEG C in temperature, intensity of illumination is 1800lx, light application time treats armpit to be cultivated 25~30 days under conditions of 10 hours/day
When bastem portion 1~2cm of diameter callus occurs and bears 5~8 3~4cm adventitious bud, then adventitious bud is cut, in phase
It is placed into the proliferated culture medium of same recipe and is transferred with the conditions of, you can the big wood paint in mao dam is carried out fast-propagation, its
Proliferation times are 6.48 times；
(5) culture of rootage：Prescription of rooting medium is：1/2MS+0.5mg/L IBA+0.5mg/L NAA+30g/L sucrose+
5.0g/L activated carbon+7.0g/L agar, pH is 5.8~6.0；In early August, 2014, propagation seedling high 2~3cm is erected and inserts in life
On root culture medium, it is 24 ± 2 DEG C to be placed in temperature, and intensity of illumination is 1800lx, and light application time is under conditions of 11~12 hour/day
Culture 25~30 days, treats that plant height reaches 3~4cm, root 1~2cm of length, that is, obtains the rooted seedling of the big wood paint in mao dam, and its rooting rate reaches
To more than 99%, 4~5 roots of average every plant of band；
(6) hardening is with transplanting：In mid-September, 2014, the bottle seedling that will take root is positioned under the conditions of natural lighting, room temperature 24 ± 2
DEG C warm canopy in carry out close lid hardening, take out rooted seedling after 7 days, the culture medium of root cleaned up, with 80% Bravo soak
After root 5min, it is peat soil to transplant to substrate composition：Perlite=3：In 2 plastic basket, sufficient root water is drenched, subenvironment phase is kept
To humidity more than 80%, relative humidity can be gradually reduced after 7 days, matrix moistening is kept, control shading rate is in 60%~70%, temperature
The seedling survived is transplanted into discrete phase after 26 ± 2 DEG C, 15~20 days of degree to educate with, a diameter of 10cm of rim of a cup degradable non-woven paper
In seedling bag, conserve 3 months, height of seedling moves to nursery lot up to 30cm or so and cultivates seedling.Make tissue culture shoot survival percent through counting this method
It is maintained at more than 90%.
The materials mode of explant through testing the application can make the induction differentiation rate of adventitious bud reach more than 85%.
Claims (1)
1. the tissue cultivation rapid breeding method of the big wood paint in a kind of mao of dam, it is characterised in that：Methods described comprises the following steps：
(1) materials of explant：The fresh axillary bud and terminal bud that annual the first tenday period of a month in May pluck the big wood paint in 5 years excellent hair dams of life are used as training
Support material；
(2) disinfecting process：The branch of harvesting is removed into blade, plant surface is cleaned with water with sterilizing banister brush, pollutant is removed, with stream
Water rinses 2h, then with after aseptic water washing 3~5 times, cuts into the segment with axillary-bud or top-bud and be positioned in sterile petri dish,
It is quickly moved in superclean bench and carries out disinfection, is aseptically soaked with 75% ethanol after 25s, be put into containing 0.01~
In 0.1% mercuric chloride thimerosal of 0.015% Tween-80,10min is gently agitated for, then with after aseptic water washing 5~8 times, by outside
Implant is placed in standby after suck dry moisture on sterilized filter paper；
(3) Fiber differentiation：The wound part of axillary bud and terminal bud contact sterilizing liquid is cut off into 3~4mm, it is perpendicular to insert in inducing culture
On, temperature is placed in carry out light culture under the conditions of 24 ± 2 DEG C, and culture is transferred in the inducing culture newly prepared for 1~2 day again, instead
Intensity of illumination is positioned over after multiple 3 times for 1800lx, light application time is illumination cultivation 20~25 under conditions of 11~12 hours/day
My god, wait to grow 3~4cm sproutings；
The formula of described inducing culture is：B5+1.0mg/L6-BA+0.1mg/LNAA+30.0g/L sucrose+5.0g/L activity
Charcoal+7.0g/L agar+0.5g/L penicillin, pH value is 5.8~6.0；
(4) Multiplying culture：Sprouting is cut from explant, its Bock Altitude is 2~3cm, 2~3 blades, is erected and inserted in
On proliferated culture medium, it is 24 ± 2 DEG C to be placed in temperature, and intensity of illumination is 1800lx, and light application time is training under conditions of 10 hours/day
Support 25~30 days, when there is 1~2cm of diameter callus in axillary bud base portion and bears 5~8 3~4cm adventitious bud, then will
Adventitious bud is cut, and is transferred in the proliferated culture medium that same recipe is placed under the same conditions, carries out the big wood paint in mao dam
Fast-propagation, its proliferation times are 6~8 times；
The proliferation culture medium formula is：MS+1.0mg/L6-BA+0.5mg/LNAA+30.0g/L sucrose+5.0g/L activated carbons+
7.0g/L agar, pH is 5.8~6.0, wherein, NH in the proliferation culture medium formula MS4NO3Concentration is the 1/ of standard medium
2；
(5) culture of rootage：Propagation seedling high 2~3cm is erected and inserted on root media, temperature is placed in for 24 ± 2 DEG C, illumination is strong
Spend for 1800lx, light application time is to cultivate 25~30 days under conditions of 11~12 hours/day, treats that plant height reaches 3~4cm, root length 1
~2cm, more than 4 number/strains of taking root, that is, obtain the rooted seedling of the big wood paint in mao dam, to wait to transplant；
The prescription of rooting medium is：1/2MS+0.5mg/LIBA+0.5mg/LNAA+30g/L sucrose+5.0g/L activated carbons+
7.0g/L agar, pH is 5.8~6.0；
(6) hardening is with transplanting：The bottle seedling that will take root is positioned under the conditions of natural lighting, carries out closing lid in the warm canopy of 24 ± 2 DEG C of room temperature
Hardening, rooted seedling is taken out after 7 days, and the culture medium of root is cleaned up, and is soaked with 80% Bravo after root 5min, is transplanted to matrix
Match as peat soil：Perlite=3：In 2 plastic basket, sufficient root water is drenched, subenvironment relative humidity more than 80%, 7 days is kept
After can gradually reduce relative humidity, keep matrix moistening, control shading rate is in 60%~70%, 26 ± 2 DEG C, 15~20 days of temperature
The seedling survived is transplanted to discrete phase with, a diameter of 10cm of rim of a cup degradable non-woven paper Seedling bag afterwards, 2~3 are conserved
Month, height of seedling moves to nursery lot up to 30cm and cultivates seedling.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
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