CN103975856A - Tissue-culture rapid propagation method for inducing regeneration of liquidamba formosana hance embryoid - Google Patents
Tissue-culture rapid propagation method for inducing regeneration of liquidamba formosana hance embryoid Download PDFInfo
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Abstract
The invention provides a tissue-culture rapid propagation method for inducing regeneration of a liquidamba formosana hance embryoid, belonging to the field of forest tissue culture breeding technology. The propagation method mainly comprises the following steps: by taking the bud and stem sections of liquidamba formosana hance as an explant, inducing fixed buds to be germinated, then inducing to produce the embryoid by taking sterile bud leaves, petioles and stems as materials, and then inducing the embryoid to generate regenerated plants for seedling exercising and domestication, and transplanting and potting. Batch production is carried out by adopting the rapid propagation technology of the liquidamba formosana hance, the method is simple and convenient, the regeneration period is short, the propagation coefficient is high, the transplanting survival rate is high, the seedlings emerge orderly and consistently so as to facilitate maintenance and management, the manpower and land resources can be saved, the production cost is low, the regenerated plants can be continuously obtained without the limit of seasons, and the method can be used for industrial production of nursery grown plants.
Description
Technical field
The present invention relates to a kind of tissue culture quick propagation culturing method of inducing sweetgum somatic embryogenesis, belong to By Tissue Culture of Trees raising technology field.
Background technology
Sweetgum is the color leaf indigenous tree species of China, and leaf look is with seasonal variation, and spring and summer leaf look light green, and leaf look had by Huang and changed in quality gradually for scarlet autumn, colorful, was a kind of good garden landscape seeds.At north America region, satin walnut is widely used in already street tree and tree is used in other greening, has obtained good greening and beautification effect.Meanwhile, sweetgum has again the title of " deserted mountain pioneer " seeds, is good ecological protection seeds.Ornamental value and economic worth thus, sweetgum is progressively extensive use in the urban afforestation of China, and demand also increases increasingly.
At present, the propagation technique of sweetgum is mainly taking seeding and seedling raising and cottage propagation as main, but these modes all have the following disadvantages: seeding and seedling raising technology emergence rate low, irregular and easily variation, the merit of preserving maternal plant is had to certain influence, the seedling cycle is long, emerges and is subject to seasonal restrictions, as emerged in enormous quantities, need a large amount of human and material resources and land resources, production cost is high; Although maternal plant merit can be well preserved in cuttage, is subject to equally seasonal effect, also has the shortcoming that production cost is high.In sum, finding method is convenient, can emerge in batches, and the sweetgum tissue-culturing rapid propagation mode that production cost is low, promotes industrialization production and have necessity.
In being permitted the thesis for the doctorate " the preliminary functional analysis of the research of sweetgum Gene Transformation System and sweetgum AGAMOUS gene " of woods, Hua Zhong Agriculture University in 2008 once mentioned the plant regeneration system of setting up sweetgum blade, be mainly taking blade as material, induction forms adventitious bud rooting hardening.Although this research has improved certain reproduction coefficient, the deficiency such as cultivation cycle is long, transplanting survival rate is low in batches.
Summary of the invention
The present invention has overcome deficiency of the prior art, by inducing embryoid body Regeneration Ways, provides a kind of cultivation simple, and reproduction coefficient is high, and the regeneration period is short, and the method for sustainable acquisition regeneration plant reduces costs simultaneously, can be used for seedling industrialization and produces.
The present invention realizes above-mentioned purpose like this:
A tissue culture and rapid propagation method of inducing sweetgum somatic embryogenesis, comprises the following steps:
(1) induction of stem section is sprouted: intercepting second to the 4th band bud tender stem segments of the raw red autumnal leaves sweetgum coppice shoot without damage by disease and insect is then material, remove blade, routine disinfection is carried out in segment, sterilization stem section is cut into 1.0-1.5cm long, is with an axillalry bud for every section, be inoculated on the medium I in triangular flask, sealing, and in temperature be at 24 ± 2 DEG C, humidity is that 75 ± 2% times, illumination are, light intensity is in the constant temperature and humidity culturing room under 1800-2000lx under 14-16h/d, cultivates 7-15d, obtains aseptic seedling;
(2) inducing embryoid body: the aseptic seedling in step (1) is divided into stem section, petiole, blade, be inoculated in the medium ii in triangular flask, sealing, at 24 ± 2 DEG C of temperature, humidity is to schedule to last respectively the dark cultivation of 10-15 days for 75 ± 2% times, again stem section, petiole, blade after dark cultivation are transferred on the medium ii I in triangular flask, sealing, schedule to last respectively the secretly cultivation for the second time of 10-25 days, to move under 300-500lx illumination condition through stem section, petiole, the blade of dark cultivation for the second time, light is cultivated 7-10d, forms bulk embryoid;
(3) embryoid induction regeneration: the bulk embryoid in step (2) is resolved into simple grain, be placed in the medium IV in triangular flask, sealing is cultivated 30-40d under 1800-2000lx illumination condition, is divided into complete regeneration plant;
(4) hardening shifts: regeneration plant bottle seedling complete in step (3) is moved on to outside culturing room, at natural daylight condition lower refining seedling 5-7d, opening bottle cap continues to transplant basin after domestication 3-5d again, transplanting medium is garden mould, leaf mould, peat soil, perlite, wherein garden mould, leaf mould, peat soil, perlitic volume ratio is 2:1:1:1, after transplanting, matrix is watered permeable, be put in PVC green house and carry out maintenance, first month, spray blade face water every day 1-2 time, relative air humidity remains on more than 80%, temperature remains 20-25 DEG C, after one month, used and become thoroughly decomposed cake fertilizer once at interval of one week, obtain sweetgum group training seedling.
Described medium I is the sucrose of TDZ, the 10-30g/L of NAA, the 0.01-0.1mg/L of 6-BA, the 0.05-0.1mg/L of WPM, 0.2-1.0mg/L, the agar powder of 2-6g/L; Wherein, the mass ratio of WPM, 6-BA, NAA, TDZ, sucrose, agar powder is 2500-3000:0.1-1.5:0.02-0.22:0.02-0.35:21000-38000:3500-7 500.
The mass ratio of the NAA of described WPM, the 6-BA of 0.5mg/L, 0.08mg/L, the TDZ of 0.04mg/L, 30g/L sucrose, 5g/L agar powder is 2700:0.85:0.15:0.22:30000:5000.
Described medium ii is the sucrose of NAA, the 10-30g/L of 6-BA, the 0.05-0.1mg/L of WPM, 0.2-1.0mg/L, the agar powder of 2-6g/L; Wherein, the mass ratio of WPM, 6-BA, NAA, sucrose, agar powder is 2500-3000:0.1-1.5:0.02-0.22:21000-38000:3500-7500.
Described medium ii is the sucrose of NAA, the 27g/L of 6-BA, the 0.02mg/L of WPM, 0.33 mg/L, the agar powder of 3.8g/L; Wherein, the mass ratio of WPM, 6-BA, NAA, sucrose, agar powder is 2850:0.46:0.17:3000:7500.
Described medium ii I is the sucrose of NAA, the 10-30g/L of TDZ, the 0.05-0.1mg/L of WPM, 0.01-0.1mg/L, the agar powder of 2-6g/L; The mass ratio of WPM, TDZ, NAA, sucrose, agar powder is 2500-3000:0.02-0.35:0.05-0.1mg/L:21000-38000:3500-7500.
Described medium ii I is the TDZ of WPM, 0.36mg/L, the sucrose of 0.03 NAA, 18g/L, the agar powder of 3.4g/L; The mass ratio of WPM, TDZ, NAA, sucrose, agar powder is 2750:0.35:0.8:29000:7500.
Described medium IV is the sucrose of WPM, 10-30g/L, the agar powder of 2-6g/L; Wherein, the mass ratio of WPM, sucrose, agar powder is 2500-3000:21000-38000:3500-7500.
Described medium IV is the sucrose of WPM, 30g/L, the agar powder of 5g/L; Wherein, the mass ratio of WPM, sucrose, agar powder is 2500:21000:3500.
The invention has the advantages that: sweetgum inducing embryoid body regeneration tissue-culturing rapid propagation practical and technical methods is easy, from explant induction to plant regeneration, transplant the hardening whole cultivation cycle tissue culture propagation mode that breaks traditions, bud induction and culture of rootage one step are completed, shorten cultivation cycle, improved reproduction coefficient, reduce man power and material's too much in group training experimentation consumption, fundamentally improve operating efficiency, saved production cost, and can reach the object of factorial seedling growth; Sweetgum embryoid induction regeneration plant, transplanting hardening survival rate is high, and the ability of plant reform of nature environment is strong, after transplanting hardening, grows rapidly, robust plant.
Brief description of the drawings
The induction of Fig. 1 sweetgum stem section is sprouted.
Fig. 2 is sweetgum blade, petiole, stem section induction acquisition embryoid.
Fig. 3 is sweetgum embryoid cluster of grains.
Fig. 4 is sweetgum embryoid induction regeneration plant.
Fig. 5 is sweetgum acclimatization and transplants.
Fig. 6 is that sweetgum is transplanted two months.
Embodiment
embodiment 1
The induction of stem section is sprouted: select to grow rapidly, be maternal plant without the good red autumnal leaves sweetgum of the proterties such as damage by disease and insect, intercepting second to the 4th band bud tender stem segments of raw red autumnal leaves sweetgum coppice shoot is then material, removes blade, and routine disinfection is carried out in segment.Sterilization stem section is cut into 1.0-1.5cm long, be with an axillalry bud for every section, be inoculated into and start medium WPM(1300g)+6-BA(0.2mg/L, 0.13g)+NAA(0.05mg/L, 0.12g)+TDZ (0.01mg/L, 0.08g)+sucrose (30g/L, 14000g)+agar powder (5g/L, 2300g), being placed in environmental condition is 24 ± 2 DEG C of temperature, humidity 75 ± 2%, illumination 14h/d, in the constant temperature and humidity culturing room of light intensity 1800-2000lx, after cultivating 10d, start to have normal bud to sprout from axillalry bud, after continuing to cultivate 12d, its inductivity reaches 100%, and average each axillalry bud all can be induced 3-4 bud, robust growth, leaf look light green (Fig. 1).
Inducing embryoid body: the aseptic seedling that (1) carrys out axillalry bud induction is divided into stem section, petiole, three kinds of materials of blade (vertical vein otch), be inoculated into medium WPM(2.5g, 1435g)+6-BA(0.3 mg/L, 0.62g)+NAA(0.01mg/L, 0.09g)+sucrose (28g/L, 12500g)+agar powder (5.2g/L, 2000g), secretly cultivate respectively (without the dark training chamber of photoenvironment, 24 ± 2 DEG C of temperature, humidity 75 ± 2%), inducing embryoid body.After dark training 10d, tender stem segments, petiole and blade all start to expand from wound.(2) by the material of step (1), it is transferred to medium WPM(1435g)+TDZ(0.3mg/L, 0.07g)+NAA(0.01 mg/L, 0.09g)+sucrose (28g/L, 12500g)+agar powder (5.2g/L, 2000g) upper, be placed in dark training chamber and carry out secondary and secretly cultivate.Tender stem segments and petiole 7d after switching, incision starts to form light green gritty texture; 10d after blade switching, incision starts to form light green gritty texture.Secretly cultivate through the secondary of 10d, the embryoid induction rate of three kinds of materials is 100%(Fig. 2), and each kind of material can produce more embryoid, and quality is loose, in good condition.(3) secondary is secretly cultivated after 25d, moves to illumination cultivation environment, and secretly the time of training can not be long at 300-500lx(for intensity control, otherwise there will be yellow, to inducing embryoid body, regeneration has certain influence), light is cultivated after 10d, and embryoid all starts to form open-textured bulk (Fig. 3).
Embryoid induction regeneration: bulk embryoid is resolved into simple grain, be put in medium WPM(1435g)+sucrose (28g/L, 12500g)+agar powder (5.2g/L, 2000g) is upper, is placed under the environment of light intensity 1800-2000lx and cultivates after 25d, embryoid starts differentiation, produce bud point by embryoid, be attended by adventive root simultaneously and produce (Fig. 4), continue to cultivate 15d, embryoid is divided into complete regeneration plant, and its regeneration plant inductivity reaches 82.3%.
Acclimatization and transplants: test-tube plantlet can complete smoothly from the procedure of adaptation to field in vitro, and the hardening before transplanting is very important.Hardening can improve the test-tube plantlet resistance of environment to external world, makes the middle transition that it enters complete open environment from a complete totally enclosed gnotobasis, and suitable hardening process can increase the resistivity of plant, most important to surviving after transplanting.
The complete regeneration plant bottle seedling of stalwartness by plant height more than 4cm moves on to outside culturing room, at natural daylight condition lower refining seedling 7d, then open bottle cap and continue domestication 5d, then transplant basin (Fig. 5), transplanting medium is garden mould, leaf mould, peat soil, perlite, wherein garden mould, leaf mould, peat soil, perlitic mass ratio is 2:1:1:1.After transplanting, matrix is watered permeablely, be put in PVC green house and carry out maintenance management.
First month after transplanting, sprays blade face water every day 1-2 time, and relative air humidity remains on more than 85%, and temperature remains on 20-25 DEG C, increases shading screen avoid high light direct projection according to weather condition, reduces warm canopy temperature simultaneously.In 3-4 week after transplanting, plant starts to grow young leaves, now suitably removes warm canopy shading screen, makes it adapt to gradually external environmental condition, and thin executing become thoroughly decomposed cake fertilizer once week about simultaneously.Transplant after two months, statistics survival rate is 81.8%(Fig. 6), the equal well-grown of all seedling, neat and consistent, is convenient to management.Grow paramount 15-20cm left and right in nutritive cube time, move to nursery, land for growing field crops and carry out field planting.
embodiment 2
The induction of stem section is sprouted: select to grow rapidly, be maternal plant without the good red autumnal leaves sweetgum of the proterties such as damage by disease and insect, intercepting second to the 4th band bud tender stem segments of raw red autumnal leaves sweetgum coppice shoot is then material, removes blade, and routine disinfection is carried out in segment.Sterilization stem section is cut into 1.0-1.5cm long, be with an axillalry bud for every section, be inoculated into and start medium WPM(1400g)+6-BA(0.8mg/L, 0.25g)+NAA(0.1mg/L, 0.04g)+TDZ (0.08mg/L, 0.038g)+sucrose (28.5g/L, 13000g)+agar powder (4.5g/L, 3500g), being placed in environmental condition is 24 ± 2 DEG C of temperature, humidity 75 ± 2%, illumination 16h/d, in the constant temperature and humidity culturing room of light intensity 1800-2000lx, after cultivating 7-10d, start to have normal bud to sprout from axillalry bud, after cultivating 10-15d, its inductivity reaches 99.8%, and average each axillalry bud all can be induced 3-4 bud, robust growth, leaf look light green.
Inducing embryoid body: the aseptic seedling that (1) carrys out axillalry bud induction is divided into stem section, petiole, three kinds of materials of blade (vertical vein otch), be inoculated into medium WPM(1500g)+6-BA(0.35 mg/L, 0.5g)+NAA(0.02mg/L, 0.045g)+sucrose (30g/L, 14000)+agar powder (5g/L, 3500g) is upper, secretly cultivates respectively (without the dark training chamber of photoenvironment, 24 ± 2 DEG C of temperature, humidity 75 ± 2%), inducing embryoid body.After dark training 12d, tender stem segments, petiole and blade all start to expand from wound.(2) by the material of step (1), it is transferred to medium WPM(1500g)+TDZ(0.45mg/L, 0.045g)+NAA(0.05 mg/L, 0.05g)+sucrose (30g/L, 12000g)+agar powder (5g/L, 3500g) upper, be placed in dark training chamber and carry out secondary and secretly cultivate.Tender stem segments and petiole 12d after switching, incision starts to form light green gritty texture; 13d after blade switching, incision starts to form light green gritty texture.Secretly cultivate through the secondary of 12d, the embryoid induction rate of three kinds of materials is 98.5%, and each kind of material can produce more embryoid, and quality is loose, in good condition.(3) secondary is secretly cultivated after 22d, moves to illumination cultivation environment, and secretly the time of training can not be long at 300-500lx(for intensity control, otherwise there will be yellow, to inducing embryoid body, regeneration has certain influence), light is cultivated after 9d, and embryoid all starts to form open-textured bulk.
Embryoid induction regeneration: bulk embryoid is resolved into simple grain, be put in medium WPM (1500g)+sucrose (30g/L, 12000g)+agar powder (5g/L, 3500g) upper,
Be placed under the environment of light intensity 1800-2000lx and cultivate after 20-25d, embryoid starts differentiation, produces bud point by embryoid, be attended by adventive root produces simultaneously, continue to cultivate 10-15d, embryoid is divided into complete regeneration plant, and its regeneration plant inductivity reaches 80.6%.
Acclimatization and transplants: test-tube plantlet can complete smoothly from the procedure of adaptation to field in vitro, and the hardening before transplanting is very important.Hardening can improve the test-tube plantlet resistance of environment to external world, makes the middle transition that it enters complete open environment from a complete totally enclosed gnotobasis, and suitable hardening process can increase the resistivity of plant, most important to surviving after transplanting.
The complete regeneration plant bottle seedling of stalwartness by plant height more than 4cm moves on to outside culturing room, at natural daylight condition lower refining seedling 5-7d, then open bottle cap and continue domestication 3-5d, then transplant basin, transplanting medium is garden mould, leaf mould, peat soil, perlite, wherein garden mould, leaf mould, peat soil, perlitic mass ratio is 2:1:1:1.After transplanting, matrix is watered permeablely, be put in PVC green house and carry out maintenance management.
First month after transplanting, sprays blade face water every day 1-2 time, and relative air humidity remains on more than 85%, and temperature remains on 20-25 DEG C, increases shading screen avoid high light direct projection according to weather condition, reduces warm canopy temperature simultaneously.In 3-4 week after transplanting, plant starts to grow young leaves, now suitably removes warm canopy shading screen, makes it adapt to gradually external environmental condition, and thin executing become thoroughly decomposed cake fertilizer once week about simultaneously.Transplant after two months, statistics survival rate is 75.8%, the equal well-grown of all seedling, and neat and consistent, is convenient to management.Grow paramount 15-20cm left and right in nutritive cube time, move to nursery, land for growing field crops and carry out field planting.
embodiment 3
The induction of stem section is sprouted: select to grow rapidly, be maternal plant without the good red autumnal leaves sweetgum of the proterties such as damage by disease and insect, intercepting second to the 4th band bud tender stem segments of raw red autumnal leaves sweetgum coppice shoot is then material, removes blade, and routine disinfection is carried out in segment.Sterilization stem section is cut into 1.0-1.5cm long, be with an axillalry bud for every section, be inoculated into and start medium WPM(1350g)+6-BA(0.5mg/L, 0.43g)+NAA(0.08mg/L, 0.08g)+TDZ (0.04mg/L, 0.11g)+sucrose (30g/L, 15000g)+agar powder (5g/L, 2500g), being placed in environmental condition is 24 ± 2 DEG C of temperature, humidity 75 ± 2%, illumination 14-16h/d, in the constant temperature and humidity culturing room of light intensity 1800-2000lx, after cultivating 8d, start to have normal bud to sprout from axillalry bud, after cultivating 11d, its inductivity reaches 100%, and average each axillalry bud all can be induced 3-4 bud, robust growth, leaf look light green.
Inducing embryoid body: the aseptic seedling that (1) carrys out axillalry bud induction is divided into stem section, petiole, three kinds of materials of blade (vertical vein otch), be inoculated into medium WPM(2850g)+6-BA(0.33 mg/L, 0.23g)+NAA(0.02mg/L, 0.085g)+sucrose (27g/L, 1500g)+agar powder (3.8g/L, 3750g)
Secretly cultivate respectively (without the dark training chamber of photoenvironment, 24 ± 2 DEG C of temperature, humidity 75 ± 2%), inducing embryoid body.After dark training 12d, tender stem segments, petiole and blade all start to expand from wound.(2) by the material of step (1), it is transferred to medium WPM(2750g)+TDZ(0.36mg/L, 0.18g)+NAA(0.03 mg/L, 0.8g)+sucrose (18g/L, 14500g)+agar powder (3.4g/L, 3750g) is upper,
Being placed in dark training chamber carries out secondary and secretly cultivates.Tender stem segments and petiole 7d after switching, incision starts to form light green gritty texture; 10d after blade switching, incision starts to form light green gritty texture.Secretly cultivate through the secondary of 14d, the embryoid induction rate of three kinds of materials is 99.5%, and each kind of material can produce more embryoid, and quality is loose, in good condition.(3) secondary is secretly cultivated after 22d, moves to illumination cultivation environment, and secretly the time of training can not be long at 300-500lx(for intensity control, otherwise there will be yellow, to inducing embryoid body, regeneration has certain influence), light is cultivated after 9d, and embryoid all starts to form open-textured bulk.
Embryoid induction regeneration: bulk embryoid is resolved into simple grain, be put in medium WPM (1250g)+sucrose (30g/L, 15000)+agar powder (5g/L, 1750g) upper,
Be placed under the environment of light intensity 1800-2000lx and cultivate after 22d, embryoid starts differentiation, produces bud point by embryoid, is attended by adventive root simultaneously and produces, and continues to cultivate 14d, and embryoid is divided into complete regeneration plant, and its regeneration plant inductivity reaches 84.6%.
Acclimatization and transplants: test-tube plantlet can complete smoothly from the procedure of adaptation to field in vitro, and the hardening before transplanting is very important.Hardening can improve the test-tube plantlet resistance of environment to external world, makes the middle transition that it enters complete open environment from a complete totally enclosed gnotobasis, and suitable hardening process can increase the resistivity of plant, most important to surviving after transplanting.
The complete regeneration plant bottle seedling of stalwartness by plant height more than 4cm moves on to outside culturing room, at natural daylight condition lower refining seedling 5-7d, then open bottle cap and continue domestication 3-5d, then transplant basin, transplanting medium is garden mould, leaf mould, peat soil, perlite, wherein garden mould, leaf mould, peat soil, perlitic mass ratio is 2:1:1:1.After transplanting, matrix is watered permeablely, be put in PVC green house and carry out maintenance management.
First month after transplanting, sprays blade face water every day 1-2 time, and relative air humidity remains on more than 85%, and temperature remains on 20-25 DEG C, increases shading screen avoid high light direct projection according to weather condition, reduces warm canopy temperature simultaneously.In 3-4 week after transplanting, plant starts to grow young leaves, now suitably removes warm canopy shading screen, makes it adapt to gradually external environmental condition, and thin executing become thoroughly decomposed cake fertilizer once week about simultaneously.Transplant after two months, statistics survival rate is 86.5%, the equal well-grown of all seedling, and neat and consistent, is convenient to management.Grow paramount 15-20cm left and right in nutritive cube time, move to nursery, land for growing field crops and carry out field planting.
Claims (9)
1. a tissue culture and rapid propagation method of inducing sweetgum somatic embryogenesis, is characterized in that, comprises the following steps:
(1) induction of stem section is sprouted: intercepting second to the 4th band bud tender stem segments of the raw red autumnal leaves sweetgum coppice shoot without damage by disease and insect is then material, remove blade, routine disinfection is carried out in segment, sterilization stem section is cut into 1.0-1.5cm long, at least be with an axillalry bud for every section, be inoculated on the medium I in triangular flask, sealing, and in temperature be at 24 ± 2 DEG C, humidity is that 75 ± 2% times, illumination are, light intensity is in the constant temperature and humidity culturing room under 1800-2000lx under 14-16h/d, cultivate 7-15d, obtain aseptic seedling;
(2) inducing embryoid body: the aseptic seedling in step (1) is divided into stem section, petiole, blade, be inoculated in the medium ii in triangular flask, sealing, at 24 ± 2 DEG C of temperature, humidity is to schedule to last respectively the dark cultivation of 10-15 days for 75 ± 2% times, again stem section, petiole, blade after dark cultivation are transferred on the medium ii I in triangular flask, sealing, schedule to last respectively the secretly cultivation for the second time of 10-25 days, to move under 300-500lx illumination condition through stem section, petiole, the blade of dark cultivation for the second time, light is cultivated 7-10d, forms bulk embryoid;
(3) embryoid induction regeneration: the bulk embryoid in step (2) is resolved into simple grain, be placed in the medium IV in triangular flask, sealing is cultivated 30-40d under 1800-2000lx illumination condition, is divided into complete regeneration plant;
(4) hardening shifts: regeneration plant bottle seedling complete in step (3) is moved on to outside culturing room, at natural daylight condition lower refining seedling 5-7d, opening bottle cap continues to transplant basin after domestication 3-5d again, transplanting medium is garden mould, leaf mould, peat soil, perlite, wherein garden mould, leaf mould, peat soil, perlitic mass ratio is 2:1:1:1, after transplanting, matrix is watered permeable, be put in PVC green house and carry out maintenance, first month, spray blade face water every day 1-2 time, relative air humidity remains on more than 80%, temperature remains 20-25 DEG C, after one month, used and become thoroughly decomposed cake fertilizer once at interval of one week, sweetgum group training seedling.
2. the tissue culture and rapid propagation method of induction sweetgum somatic embryogenesis according to claim 1, is characterized in that: medium I is the sucrose of TDZ, the 10-30g/L of NAA, the 0.01-0.1mg/L of 6-BA, the 0.05-0.1mg/L of WPM, 0.2-1.0mg/L, the agar powder of 2-6g/L; Wherein, the mass ratio of WPM, 6-BA, NAA, TDZ, sucrose, agar powder is 2500-3000:0.1-1.5:0.02-0.22:0.02-0.35:21000-38000:3500-7 500.
3. the tissue culture and rapid propagation method of induction according to claim 2 sweetgum somatic embryogenesis, is characterized in that: the mass ratio of the TDZ of the 6-BA of WPM, 0.5mg/L, the NAA of 0.08mg/L, 0.04mg/L, 30g/L sucrose, 5g/L agar powder is 2700:0.85:0.15:0.22:30000:5000.
4. the tissue culture and rapid propagation method of induction sweetgum somatic embryogenesis according to claim 1, is characterized in that: medium ii is the sucrose of NAA, the 10-30g/L of 6-BA, the 0.05-0.1mg/L of WPM, 0.2-1.0mg/L, the agar powder of 2-6g/L; Wherein, the mass ratio of WPM, 6-BA, NAA, sucrose, agar powder is 2500-3000:0.1-1.5:0.02-0.22:21000-38000:3500-7500.
5. the tissue culture and rapid propagation method of induction sweetgum somatic embryogenesis according to claim 4, is characterized in that: medium ii is the sucrose of NAA, the 27g/L of 6-BA, the 0.02mg/L of WPM, 0.33 mg/L, the agar powder of 3.8g/L; Wherein, the mass ratio of WPM, 6-BA, NAA, sucrose, agar powder is 2850:0.46:0.17:3000:7500.
6. the tissue culture and rapid propagation method of induction sweetgum somatic embryogenesis according to claim 1, is characterized in that: medium ii I is the sucrose of NAA, the 10-30g/L of TDZ, the 0.05-0.1mg/L of WPM, 0.01-0.1mg/L, the agar powder of 2-6g/L; The mass ratio of WPM, TDZ, NAA, sucrose, agar powder is 2500-3000:0.02-0.35:0.05-0.1:21000-38000:3500-7500.
7. the tissue culture and rapid propagation method of induction according to claim 6 sweetgum somatic embryogenesis, is characterized in that: medium ii I is the TDZ of WPM, 0.36mg/L, the sucrose of 0.03 NAA, 18g/L, the agar powder of 3.4g/L; The mass ratio of WPM, TDZ, NAA, sucrose, agar powder is 2750:0.35:0.8:29000:7500.
8. the tissue culture and rapid propagation method of induction sweetgum somatic embryogenesis according to claim 1, is characterized in that: medium IV is the sucrose of WPM, 10-30g/L, the agar powder of 2-6g/L; Wherein, the mass ratio of WPM, sucrose, agar powder is 2500-3000:21000-38000:3500-7500.
9. the tissue culture and rapid propagation method of induction sweetgum somatic embryogenesis according to claim 8, is characterized in that: medium IV is the sucrose of WPM, 30g/L, the agar powder of 5g/L; Wherein, the mass ratio of WPM, sucrose, agar powder is 2500:21000:3500.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105191794A (en) * | 2015-09-24 | 2015-12-30 | 广西壮族自治区药用植物园 | Method for rapid propagation of bischofia javanica based on embryogenic callus seedling development |
| CN114365691A (en) * | 2022-02-21 | 2022-04-19 | 德兴市荣兴苗木有限责任公司 | Tissue culture method of fortune purple maple |
| CN114467753A (en) * | 2022-02-21 | 2022-05-13 | 德兴市荣兴苗木有限责任公司 | Tissue culture method of silvery deer maple |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1341351A (en) * | 2001-10-18 | 2002-03-27 | 南京林业大学 | Chinese sweetgum tissue culture and quick propagation method |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1341351A (en) * | 2001-10-18 | 2002-03-27 | 南京林业大学 | Chinese sweetgum tissue culture and quick propagation method |
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| C.C.GIRI·B ETAL: "Progress in tissue culture,genetic transformation and applications of biotechnology to trees:an overview", 《TREES》 * |
| 施季森等: "中国枫香育种研究现状", 《林业科技开发》 * |
| 苏梦云: "美国枫香茎段组织培养与植株再生", 《林业科学研究》 * |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105191794A (en) * | 2015-09-24 | 2015-12-30 | 广西壮族自治区药用植物园 | Method for rapid propagation of bischofia javanica based on embryogenic callus seedling development |
| CN114365691A (en) * | 2022-02-21 | 2022-04-19 | 德兴市荣兴苗木有限责任公司 | Tissue culture method of fortune purple maple |
| CN114467753A (en) * | 2022-02-21 | 2022-05-13 | 德兴市荣兴苗木有限责任公司 | Tissue culture method of silvery deer maple |
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| CN103975856B (en) | 2016-03-02 |
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