CN106305427B - A kind of method of Amur maple tissue-culturing quick-propagation - Google Patents
A kind of method of Amur maple tissue-culturing quick-propagation Download PDFInfo
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- CN106305427B CN106305427B CN201610759194.2A CN201610759194A CN106305427B CN 106305427 B CN106305427 B CN 106305427B CN 201610759194 A CN201610759194 A CN 201610759194A CN 106305427 B CN106305427 B CN 106305427B
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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
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
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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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Abstract
The present invention provides a kind of method of Amur maple tissue-culturing quick-propagation, it is characterized in that preferably woody on the basis of, 5 years raw no disease and pests harms of Amur maple, the method given birth to coppice shoot and mature embryo then and carry out tissue-culturing rapid propagation of healthy and strong plant, category Tissue Culture of Trees raising technology field is respectively adopted.Main technical points are using the Amur maple elite plant of screening as female parent, green tape leaf stem section and mature embryo are explant then for selection, induction is sprouted, and is then induced with aseptic seedling and is produced adventitious bud, then taken root by adventitious bud inducing regeneration plant carry out hardening domestication, in transplanting basin propagation method.Produced in batches using Amur maple fast breeding technique provided by the invention, method is easy, regeneration period is short, and breeding coefficient is high, and transplanting survival rate is high, emergence neat and consistent, it is easy to maintenance management, saves manpower and land resource, production cost are low, lasting acquisition regeneration plant is not subject to seasonal restrictions, is produced available for seedling industrialization.
Description
Technical field
The present invention relates to the practical technique of Amur maple tissue-culturing quick-propagation.
Background technology
Amur maple is the big shrub of Aceraceae Acer fallen leaves or dungarunga, and for viewing and admiring, blade profile is beautiful for leaf, fruit.Autumn leaf colours are red
It is gorgeous, it is especially noticeable；The double samara pinkiness that summer has just born, very delicate, unique, being that the north is excellent views and admires green
Change seeds.The wood in addition, its timber can run business into particular one；Tender leaf is processed to be used for tea, have promote the production of body fluid to quench thirst, improving eyesight and other effects of bringing down a fever, also
It can extract substantial amounts of gallic acid (Gallic acid, Gallic Acid).Gallic acid is also known as gallic acid, Chinese gall
Acid, it is of wide application, is mainly used in medicine, dyestuff, ink manufacture, is also used as food antioxidant, preservative, METAL EXTRACTION
Agent, ultra-violet absorber, disinfectant, hemostasis astringent, developer, chemical reagent, mud fluidizing reagent and grape growth agent etc..Grind
Study carefully and show, the growth inhibition or apoptosis of tumour cell can be made using gallic acid as medicine made of raw material, is alternatively arranged as antioxidant,
With stronger antioxidation, on International Medical, it is general anti-infective medicine such as four oxygen have been synthesized using gallic acid as raw material
Woods (TXP), brodimoprim (BOP) and metioprim (MTP)；DE 2051443 (Exifone), for treating the old disturbance of consciousness；Connection
The double fat of benzene, for treating chronic hepatitis；Dilazep, worry are flat, diphenamilat, for angina pectoris, heart failure, myocardial infarction
Deng；Logical board ester, for treating cerebral thrombus；Troxipide, for treating gastric ulcer；Gram hat acid, for treating miocardial infarction；Not yet
Gallate-based antimony sodium, for treating blood fluke；Ellagic acid, for anti-cancer and anticancer.As development in science and technology and foreign trade constantly expand, need
The amount of asking will be increasing.But current Amur maple there are no natural good variety population, mostly inlay scattered distribution, predatoriness is adopted in addition
Receive, in shape of gradually endangering.
At present, the reproduction technique of Amur maple is mainly based on seeding and seedling raising, but these modes all have the following disadvantages：
Seeding and seedling raising technology emergence rate is low, irregular and easily variation, has certain influence, seedling cycle to the merit for preserving maternal plant
Long, emergence is subject to seasonal restrictions, such as in high volume emerged, and needs a large amount of human and material resources and land resource, production cost height.To sum up institute
State, finding method is convenient, can emerge in batches, the low Amur maple tissue-culturing rapid propagation mode of production cost, promotes industrialization production great
Necessity.
Once being mentioned in the gorgeous Master's thesis in Northeast Forestry University Lee sea in 2007 uses seed to establish tea bar for explant
The Tissue Culture Regeneration System of maple；Gansu Agriculture University's Li Yan rocks are mentioned in master's thesis with seedling stem section within 2007
Tissue culture rapid propagation system is established for explant；Northeast Forestry University Life Science College Dong Jie in 2008 etc. is outer using Amur maple cotyledon
Implant tissue culture obtains gallic acid.Several researchs above are carried out for the purpose of extracting Amur maple gallic acid, in heredity
The stable aspect of shape is without progress systematic research, therefore its Ornamental value not yet develops.
The content of the invention
Merit of the invention by screening maternal plant, lured using the regeneration of stem section evoking adventive bud and mature seed embryo culture
The method for leading plant regeneration, overcome the characteristics of aseptic seedling browning in the prior art, growth coefficient are low and cultivation cycle is long, there is provided
A kind of culture is simple, and breeding coefficient is high, and the regeneration period is short, sustainable to obtain the method for regeneration plant, while reduces cost, can
Produced for seedling industrialization.
The practical approach of Amur maple tissue-culturing quick-propagation
The selection of explant：Two-year phenological period observation is carried out to Amur maple, records the rudiment per seedlings in detail
Phase, florescence, fruiting period, samara color, red autumnal leaves phase, leaf fall period, pest and disease damage situation, filter out the healthy and strong plant of Amur maple of no disease and pests harm
Strain；
Stem section induction is sprouted：Learnt from else's experience the plant no disease and pests harm coppice shoot of screening, interception gives birth to first of coppice shoot to the then
Four band bud tender stem segmentses are experiment material, remove blade, and segment carries out routine disinfection, and sterilization stem section is cut into 1.5-2.0cm length,
A pair of axillary buds of every section of band, are inoculated in primary culture medium, are cultivated 12-14 days, are made each 1-2 bud of axillary bud deriving, and leaf color is tender
It is green, the aseptic seedling of robust growth；Described primary culture medium is MS+6-BA0.5-1.0mg/L+NAA0.05-0.1mg/L+
AC0.5-1.0g/L+PVP100-200mg/L+ sucrose 30g/L+ agar powder 4.5g/L, its culture environment condition be temperature 23 ±
2 DEG C, humidity 60 ± 2%, illumination 12-14h/d, light intensity 1500-1800lx constant temperature and humidity culturing room in.
Embryo culture induction is sprouted：The merit Amur maple screened of learning from else's experience is female parent, using morphological maturity seed, is removed
Fruit wing, embathed 15 minutes with washing powder, flowing water rinses 2 hours, then is carried out disinfection in super bacterium workbench.Disinfecting process is：First use
75% alcohol sterilizes 1 minute, pours out with sterile water wash 2 times, then is sterilized 30 minutes with 0.1% mercuric chloride, pours out with sterile water wash 4
It is secondary, first time Rapid Cleaning, after three times every time cleaning 3 minutes, the method can thoroughly clean mercuric chloride residue, avoid injury kind
Embryo.The seed that will be disinfected, exosper is removed, is inoculated into the culture medium of embryo culture, cultivated 2 weeks, radicle and cotyledon progressively produce
It is raw, and then form complete aseptic seedling；The culture medium of described embryo culture is MS+GA31.0-3.0mg/L+AC0.5-1.0g/L+
PVP 100-200mg/L+ sucrose 30g/L+ agar powder 4.5g/L, its culture environment condition be 23 ± 2 DEG C of temperature, humidity 60 ±
2%th, in illumination 12-14h/d, light intensity 1500-1800lx constant temperature and humidity culturing room.After primary vaccination three days, for the first time turn
Connect, once, culture medium is constant for switching in 20 days thereafter.This method effectively controls browning, while can adsorb kind by way of tube
Suppress the inhibiting substances of its sprouting in embryo, promote embryo germination；
Multiplying culture：Sprouting or the aseptic seedling of embryo culture induction sprouting induction is induced to be inoculated into Multiplying culture stem section
Cultivate, cultivate 12-16 days on base, obtain dissolving the seedling of sprouting from base portion and axillary bud punishment；Proliferated culture medium is MS+6-
BA0.05-0.1mg/L+NAA0.1-0.2mg/L+TDZ0.5-1.0mg/L+AC0.5-1.0g/ L+ PVP100-200mg/L+ sugarcanes
Sugared 30g/L+ agar powders 4.5g/L, its culture environment condition are 23 ± 2 DEG C of temperature, humidity 60 ± 2%, illumination 12-14h/d, light intensity
In 1500-1800lx constant temperature and humidity culturing room.
Strengthening seedling and rooting culture：When sprouting grows to 2-3cm, the simple bud of robust growth is cut, is inoculated into root media
Middle culture, after cultivating 25-30d, there is adventitious root generation, form complete plant；Root media is MS+IBA0.5-1.0mg/L
+ AC0.5-1.0g/L+PVP100-200mg/L+ sucrose 20g/L+ agar powder 4.5g/L, its culture environment condition are temperature 23 ± 2
DEG C, in humidity 60 ± 2%, illumination 12-14h/d, light intensity 1500-1800lx constant temperature and humidity culturing room.This method overcomes existing
Technical deficiency, a step complete strong sprout and culture of rootage, save cultivation cycle, and obtain the rooted seedling of stalwartness.
Acclimatization and transplantses：Take root bottle seedling of the plant height in more than 4cm is moved on to outside culturing room, in natural scattered light condition lower refining seedling
3~7d, 5~7d is tamed in transplanting front opening bottle cap, take out aseptic seedling, the culture medium of cleaning plant root attachment, transplanted
Upper basin, transplanting medium are leaf mould：Peat soil：Perlite 1：1：1, with 1000 times of liquid of carbendazim matrix poured after transplanting it is permeable,
It is put in PVC warmhouse booths and carries out maintenance management；Root media is MS+IBA0.5-1.0mg/L+AC0.5-1.0g/L+
PVP100-200mg/L+ sucrose 20g/L+ agar powder 4.5g/L, its culture environment condition be 23 ± 2 DEG C of temperature, humidity 60 ± 2%,
Illumination 12-14h/d, light intensity 1500-1800lx constant temperature and humidity culturing room in.
Maintenance management：The abundant place of scattering light is placed on after transplanting, 1~3 week after transplanting, is sprayed water 1~2 time daily, air
Relative humidity is maintained at more than 60%, and after transplanting one month, plant starts to grow young leaves, now applies decomposed cake fertilizer every two weeks thin
Once, once, after two months, growth of seedling is good for transplanting for the prevention and control of plant diseases, pest control, when paramount 15~20cm is grown in nutritive cube, moves
It is colonized to land for growing field crops.
The advantages of Amur maple tissue-culturing quick-propagation practical technique
Amur maple tissue-culturing quick-propagation practical and technical methods are easy, and induction plant regeneration is screened from explant, move
Plant the whole cultivation cycle of hardening and break existing tissue culture propagation mode, while plant merit is kept, shorten culture week
Phase, breeding coefficient is improved, reduce the consumption of man power and material excessive in tissue culture experimentation, fundamentally improve work
Make efficiency, save production cost, and the purpose of industrial seedling rearing can be reached；
Amur maple leaf regeneration plant, transplanting hardening survival rate is high, and the ability of plant reform of nature environment is strong, transplants hardening
Growth is rapid afterwards, robust plant.
Brief description of the drawings
Fig. 1 is that the induction of the Amur maple stem section of embodiment 1 is sprouted.
Fig. 2 is the Amur maple Multiplying culture of embodiment 1.
Fig. 3 is the Amur maple strengthening seedling and rooting culture of embodiment 1.
Fig. 4 is the Amur maple acclimatization and transplantses of embodiment 1.
Fig. 5 is that the Amur maple of embodiment 1 transplants half a year growing state.
Fig. 6 is that the Amur maple embryo culture of embodiment 2 induces radicle to produce.
Fig. 7 is that the Amur maple embryo culture of embodiment 2 induces sprouting to form whole seedlings.
Fig. 8 is the Amur maple Multiplying culture of embodiment 2.
Fig. 9 is the Amur maple strengthening seedling and rooting culture of embodiment 2.
Embodiment
Embodiment 1
The Amur maple that the selection of explant is introduced a fine variety to Three Gorges nursery carries out two-year phenological period observation, records in detail
Budding period, florescence, fruiting period, samara color, red autumnal leaves phase, leaf fall period, pest and disease damage per seedlings etc., will be above-mentioned according to appreciation effect
Parameter is compared, to filter out optimal plant.
The elite plant no disease and pests harm coppice shoot for screening of learning from else's experience is sprouted in stem section induction, and first of coppice shoot is given birth in interception then
It is experiment material to the 4th band bud tender stem segmentses, removes blade, segment carries out routine disinfection.Sterilization stem section is cut into 1.5-
2.0cm grows, and a pair of axillary buds of every section of band, is inoculated into primary culture medium MS+6-BA0.8mg/L+NAA0.06mg/L+AC0.8g/L+
PVP150mg/L+ sucrose 30g/L+ agar powder 4.5g/L, environmental condition is placed in as 23 ± 2 DEG C of temperature, humidity 60 ± 2%, illumination
12-14h/d, light intensity 1500-1800lx constant temperature and humidity culturing room in, begin with normal bud after cultivating 5-7d at axillary bud and sprout,
Its inductivity is up to 100% after cultivating 12-14d, and average each axillary bud can induce 1-2 bud, and leaf color is light green, robust growth（Figure
1）.
The aseptic seedling that Multiplying culture carrys out axillary bud deriving, which is inoculated on proliferated culture medium, cultivates, proliferated culture medium MS+
6-BA0.08mg/L+NAA0.15mg/L+TDZ0.8mg/L+AC0.8g/L+PVP150mg/L+ sucrose 30g/L+ agar powders 4.5g/
L, condition of culture are same as above.Base portion slightly expanded before this after inoculation 7d, in yellow green, subsequently formed lumps callus, 2 weeks,
Start to dissolve sprouting from base portion and axillary bud punishment（Fig. 2）, it is 5-7 to count its average coefficient of proliferation after two months.
The simple bud of robust growth is cut when sprouting grows to 2-3cm, is inoculated into root media by strengthening seedling and rooting culture
Middle culture, root media are MS+IBA0.8mg/L+AC0.8g/L+PVP150mg/L+ sucrose 20g/L+ agar powder 4.5g/L,
Condition of culture is same as above.After cultivating 25-30d, there is adventitious root generation（Fig. 3）, so as to form complete plant, its rooting rate reaches
100%。
Acclimatization and transplantses test tube seedling can be smoothly completed to the adaptation process in field out of test tube, and the hardening before transplanting is very
Important.Hardening can improve resistance of the test tube seedling to external environment, it is entered from a completely enclosed gnotobasis
The middle transition of open environment, suitable hardening process completely, the resistivity of plant can be increased, to surviving to pass after transplanting
It is important.
Take root bottle seedling of the plant height in more than 4cm is moved on to outside culturing room, in natural scattered light condition 3~7d of lower refining seedling, so
5~7d is tamed in transplanting front opening bottle cap afterwards, aseptic seedling is further taken out, the culture medium of cleaning plant root attachment, is finally moved
Basin in cultivation（Fig. 4）, transplanting medium is leaf mould：Peat soil：Perlite 1：1：1.1000 times of liquid of carbendazim are used after transplanting by matrix
Pour it is permeable, be put in PVC warmhouse booths carry out maintenance management.
Influence from matrix to transplanting survival rate considers that Amur maple category compares the plant for liking well-drained sandy loam,
Therefore selection gas permeability is preferable, and the more leaf mould of contained nutriment, peat soil and perlite are matched, ratio 1:1:
1 mixing is mixed thoroughly as transplanting medium.Amur maple is a kind of heliophilous species simultaneously, compares light, scattering light is placed on after transplanting
Abundant place.1~3 week after transplanting, spray water 1~2 time daily, relative air humidity is maintained at more than 60%, but does not exceed 90%, temperature
Degree is maintained at 20 DEG C or so（Because under the high temperature conditions, relative humidity is higher than 90%, rotten seedling phenomenon easily occurs.）.According to weather feelings
Condition adds shading screen to avoid strong light direct beam, reduces warm canopy temperature.After transplanting one month, plant starts to grow young leaves, now every two
Week it is thin apply decomposed cake fertilizer once, the prevention and control of plant diseases, pest control is once.After two months, statistics survival rate is 90%, and all seedling grow for transplanting
Well.When paramount 20cm is grown in nutritive cube, move to land for growing field crops and be colonized（Fig. 5）.
Embodiment 2
The Amur maple that the selection of explant is introduced a fine variety to Three Gorges nursery carries out two-year phenological period observation, records in detail
Budding period, florescence, fruiting period, samara color, red autumnal leaves phase, leaf fall period, pest and disease damage per seedlings etc., will be above-mentioned according to appreciation effect
Parameter is compared, to filter out optimal plant.
The merit Amur maple that screening of learning from else's experience is sprouted in embryo culture induction is female parent, using morphological maturity seed, is removed
Fruit wing, embathed 15 minutes with washing powder, flowing water rinses 2 hours, then is carried out disinfection in super bacterium workbench.Disinfecting process is：First use
75% alcohol sterilizes 1 minute, pours out with sterile water wash 2 times, then is sterilized 30 minutes with 0.1% mercuric chloride, pours out with sterile water wash 4
It is secondary, first time Rapid Cleaning, after three times every time cleaning 3 minutes, the method can thoroughly clean mercuric chloride residue, avoid injury kind
Embryo.The seed that will be disinfected, exosper is removed, is inoculated on the culture medium of embryo culture and cultivates, culture medium MS+GA31.0-
3.0mg/L+AC0.5-1.0g/L+PVP 100-200mg/L+ sucrose 30g/L+ agar powder 4.5g/L, environmental condition is placed in as temperature
Degree 23 ± 2 DEG C, humidity 60 ± 2%, illumination 12-14h/d, light intensity 1500-1800lx constant temperature and humidity culturing room in, cultivate 2 weeks,
Radicle and cotyledon progressively produce（Fig. 6）, and then form complete aseptic seedling（Fig. 7）.After primary vaccination three days, for the first time turn
Connect, once, culture medium is constant for switching in 20 days thereafter.This method effectively controls browning, while can adsorb kind by way of tube
Suppress the inhibiting substances of its sprouting in embryo, promote embryo germination；
The aseptic seedling that Multiplying culture carrys out axillary bud deriving, which is inoculated on proliferated culture medium, cultivates, proliferated culture medium MS+
6-BA0.08mg/L+NAA0.15mg/L+TDZ0.8mg/L+AC0.8g/L+PVP150mg/L+ sucrose 30g/L+ agar powders 4.5g/
L, condition of culture are same as above.Base portion slightly expanded before this after inoculation 7d, in yellow green, subsequently formed lumps callus, 2 weeks,
Start to dissolve sprouting from base portion and axillary bud punishment（Fig. 8）, it is 5-7 to count its average coefficient of proliferation after two months.
The simple bud of robust growth is cut when sprouting grows to 2-3cm, is inoculated into root media by strengthening seedling and rooting culture
Middle culture, root media are MS+IBA0.8mg/L+AC0.8g/L+PVP150mg/L+ sucrose 20g/L+ agar powder 4.5g/L,
Condition of culture is same as above.After cultivating 25-30d, there is adventitious root generation, so as to form complete plant, its rooting rate is up to 100%（Figure
9）.
Acclimatization and transplantses test tube seedling can be smoothly completed to the adaptation process in field out of test tube, and the hardening before transplanting is very
Important.Hardening can improve resistance of the test tube seedling to external environment, it is entered from a completely enclosed gnotobasis
The middle transition of open environment, suitable hardening process completely, the resistivity of plant can be increased, to surviving to pass after transplanting
It is important.
Take root bottle seedling of the plant height in more than 4cm is moved on to outside culturing room, in natural scattered light condition 3~7d of lower refining seedling, so
5~7d is tamed in transplanting front opening bottle cap afterwards, aseptic seedling is further taken out, the culture medium of cleaning plant root attachment, is finally moved
Basin in cultivation, transplanting medium are leaf mould：Peat soil：Perlite 1：1：1.Matrix is irrigated with 1000 times of liquid of carbendazim after transplanting
Water, it is put in PVC warmhouse booths and carries out maintenance management.
Influence from matrix to transplanting survival rate considers that Amur maple category compares the plant for liking well-drained sandy loam,
Therefore selection gas permeability is preferable, and the more leaf mould of contained nutriment, peat soil and perlite are matched, ratio 1:1:
1 mixing is mixed thoroughly as transplanting medium.Amur maple is a kind of heliophilous species simultaneously, compares light, scattering light is placed on after transplanting
Abundant place.1~3 week after transplanting, spray water 1~2 time daily, relative air humidity is maintained at more than 60%, but does not exceed 90%, temperature
Degree is maintained at 20 DEG C or so（Because under the high temperature conditions, relative humidity is higher than 90%, rotten seedling phenomenon easily occurs.）.According to weather feelings
Condition adds shading screen to avoid strong light direct beam, reduces warm canopy temperature.After transplanting one month, plant starts to grow young leaves, now every two
Week it is thin apply decomposed cake fertilizer once, the prevention and control of plant diseases, pest control is once.After two months, statistics survival rate is 90%, and all seedling grow for transplanting
Well.When paramount 20cm is grown in nutritive cube, move to land for growing field crops and be colonized.
Claims (1)
- A kind of 1. method of Amur maple tissue-culturing quick-propagation, it is characterised in that comprise the following steps：The selection of explant：Two-year phenological period observation is carried out to Amur maple, records budding period, the flower of every seedlings in detail Phase, fruiting period, samara color, red autumnal leaves phase, leaf fall period, pest and disease damage situation, filter out no disease and pests harm, healthy and strong Amur maple plant；Stem section induction is sprouted：Learnt from else's experience the plant no disease and pests harm coppice shoot of screening, interception gives birth to first to the 4th of coppice shoot then Band bud tender stem segmentses are experiment material, remove blade, and segment carries out routine disinfection, and sterilization stem section is cut into 1.5-2.0cm and grown, every section A pair of axillary buds of band, are inoculated in primary culture medium, are cultivated 12-14 days, are made each 1-2 bud of axillary bud deriving, and leaf color is light green, raw Long healthy and strong aseptic seedling；Described primary culture medium is MS+6-BA0.5-1.0mg/L+NAA0.05-0.1mg/L+AC0.5- 1.0g/L+PVP100-200mg/L+ sucrose 30g/L+ agar powder 4.5g/L, its culture environment condition are 23 ± 2 DEG C of temperature, humidity 60 ± 2%, in illumination 12-14h/d, light intensity 1500-1800lx constant temperature and humidity culturing room；Embryo culture induction is sprouted：The merit Amur maple screened of learning from else's experience is female parent, using morphological maturity seed, removes fruit Wing, embathed 15 minutes with washing powder, flowing water rinses 2 hours, then is carried out disinfection in super bacterium workbench, the seed that will be disinfected, goes Except exosper, it is inoculated into the culture medium of embryo culture, cultivates 2 weeks, radicle and cotyledon progressively produces, and then are formed complete sterile Seedling, primary vaccination carry out first time switching, once, culture medium is constant, and this method effectively controls brown for switching in 20 days thereafter after three days Change, while the inhibiting substances for suppressing its sprouting in embryo can be adsorbed by way of tube, promote embryo germination, the training of described embryo Foster culture medium is MS+GA31.0-3.0mg/L+AC0.5-1.0g/L+PVP100-200mg/L+ sucrose 30g/L+ agar powders 4.5g/L, its culture environment condition are 23 ± 2 DEG C of temperature, humidity 60 ± 2%, illumination 12-14h/d, light intensity 1500-1800lx In constant temperature and humidity culturing room；Multiplying culture：Sprouting or the aseptic seedling of embryo culture induction sprouting induction is induced to be inoculated on proliferated culture medium stem section Culture, cultivate 12-16 days, obtain dissolving new sprout from base portion and axillary bud punishment, described proliferated culture medium is MS+6- BA0.05-0.1mg/L+NAA0.1-0.2mg/L+TDZ0.5-1.0mg/L+ AC 0.5-1.0g/L +PVP100-200mg/L+ Sucrose 30g/L+ agar powder 4.5g/L, its culture environment condition are 23 ± 2 DEG C of temperature, humidity 60 ± 2%, illumination 12-14h/d, light In strong 1500-1800lx constant temperature and humidity culturing room；Strengthening seedling and rooting culture：When sprouting grows to 2-3cm, the simple bud of robust growth is cut, is inoculated into root media and trains Support, after cultivating 25-30d, there is adventitious root generation, form complete healthy and strong plant, described root media is MS+IBA0.5- 1.0mg/L+AC0.5-1.0g/L+PVP100-200mg/L+ sucrose 20g/L+ agar powder 4.5g/L, its culture environment condition are temperature Degree 23 ± 2 DEG C, humidity 60 ± 2%, illumination 12-14h/d, light intensity 1500-1800lx constant temperature and humidity culturing room in；Acclimatization and transplantses：Take root bottle seedling of the plant height in more than 4cm is moved on to outside culturing room, natural scattered light condition lower refining seedling 3~ 7d, 5~7d is tamed in transplanting front opening bottle cap, take out aseptic seedling, the culture medium of cleaning plant root attachment, transplanted Basin, transplanting medium are leaf mould：Peat soil：Perlite 1：1：1, with 1000 times of liquid of carbendazim matrix poured permeable after transplanting, put Maintenance management is carried out in PVC warmhouse booths；Maintenance management：The abundant place of scattering light is placed on after transplanting, 1~3 week after transplanting, is sprayed water 1~2 time daily, air is relative Humidity is maintained at more than 60%, and after transplanting one month, plant starts to grow young leaves, now applies decomposed cake fertilizer once every two weeks thin, Once, after two months, growth of seedling is good for transplanting for the prevention and control of plant diseases, pest control, when paramount 15~20cm is grown in nutritive cube, moves to big Field nursery is colonized.
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| CN112806264B (en) * | 2021-01-26 | 2023-03-31 | 中国科学院华南植物园 | Tissue culture rapid propagation method for Acer catalpa bungei |
| CN114431154B (en) * | 2022-04-08 | 2022-06-28 | 东北林业大学 | Method for asexual propagation through acer nikoense dormant buds |
| CN114711143B (en) * | 2022-04-21 | 2023-08-08 | 东北林业大学 | Method for asexually and rapidly propagating Acer ginnala Maxim seedlings through tissue culture |
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