CN108901838A - White palm nursery propagation method - Google Patents

Abstract

The invention discloses white palm nursery propagation methods, including：Vernalization, Fiber differentiation, shoot proliferation culture, tissue culture seedling rooting, first by it is white the palm seed be soaked into after liquor natrii hypochloritis sterilizes containing ethyl alcohol, N- benzylquininium chloride aqueous solution in vernalization, rudiment is carried out after taking-up, young shoot is placed in MS culture medium and carries out Fiber differentiation, bud section is taken to be inoculated with progress shoot proliferation culture in MS culture medium, the seedling that culture obtains is placed in 1/2MS culture medium+1/2B5 culture medium and carries out culture of rootage, can be transplanted after taking seedling hardening.It has the beneficial effect that：This nursery propagation method simple process, cheap environmental protection, the well developed root system of white palm seedling, root Numerous, diameter are coarse, high survival rate, dialogue palm presprouting of seeds can accelerate its protease to secrete, and accelerate the respiratory rate, glycolytic cycle and mitochondria activity of the white palm, white palm seed is promoted to revive from suspend mode ahead of time.

Description

White palm nursery propagation method

Technical field

The present invention relates to botanical seedling culturing fields, more particularly, to white palm nursery propagation method.

Technical background

The white palm, scientific name crane taro, Latin are entitledSpathiphyllum kochii Engl. & K.Krause, Araeceae Perennial herb.30~40 centimetres of plant height, acaulescence or stem are short and small, and leaf is oval or nearly lanceolar, has apparent middle arteries and petiole, deep Green.Spring and summer is bloomed, and spathe is big and significant, is higher by blade face, white or micro- green, spadix milk yellow.Originate in America heat Band area, all over the world cultivation extensively.The white palm is very beautiful when blooming, and is also that excellent indoor pot sees leaf plant when not blooming Object.It is the indoor pot flowers of a new generation.The white palm can filter indoor exhaust air, have to ammonia, acetone, benzene and formaldehyde certain Cleaning efficacy is also used as cut-flower.The white pyrophilous high humidity of the palm, it is also more shade tolerant.Blade is larger, more sensitive to humidity, is afraid of Strong light is exposed to the sun, Shading in Summer 60~70%, but long-term illumination is insufficient, then is not easy to bloom.Soil is with fertile, abundant containing humus Loam is preferably.Growing thermophilic is 22~28 DEG C, and 3~September is with 24~30 DEG C, and September to March in next year is 18~21 DEG C, winter temperature Not less than 14 DEG C, temperature is obstructed lower than 10 DEG C of plant strain growths, and blade is vulnerable to freeze injury.The white palm of potting is in transporting procedures, temperature control At 13~16 DEG C, relative humidity 80~90% is resistant to dark surrounds 30 days system.

Summary of the invention

The purpose of the present invention is to provide white palm nursery propagation methods, this nursery propagation method simple process is cheap environmentally friendly, The well developed root system of white palm seedling, root Numerous, diameter are coarse, high survival rate, and dialogue palm presprouting of seeds can accelerate its albumen Enzyme secretion, accelerates the respiratory rate, glycolytic cycle and mitochondria activity of the white palm, white palm seed is promoted to revive from suspend mode ahead of time It wakes up.

Aiming at the problem that mentioning in background technique, the technical solution taken is the present invention：

White palm nursery propagation method, including：Vernalization, Fiber differentiation, shoot proliferation culture, tissue culture seedling rooting, specifically include following Step：

Vernalization：Picking shape is full, size is close to intact white palm seed, with 1~2% liquor natrii hypochloritis disinfection treatment 3~ 5min, distilled water flushing is clean, is put into 2~3h of vernalization in 0~1 DEG C of ice bath pot water, contains 10~12g/L ethyl alcohol, 15 in water ~18mg/LN- benzylquininium chloride pads one layer of filter paper and double gauze in culture dish, is put into seed to inhale paper suck dry moisture It is placed in incubator and carries out rudiment；Incubator temperature is 24~26 DEG C, and light application time is 10~12h, intensity of illumination 2000 ~2400Lux, daily spray water stop sprinkling after germination；Micro N- benzylquininium chloride is added in vernalization aqueous solution It can help to vernalization with suitable ethyl alcohol, under the action of ethanol in proper amount, N- benzylquininium chloride can after entering white palm seed To stimulate the related gene expression of the white palm, accelerates its protease to secrete, improve its proteinase activity, while N- benzylquininium chlorination Object can also accelerate the respiratory rate of the white palm, and further strengthen glycolytic cycle and mitochondria activity, enhance soluble sugar Secretion, promotes white palm seed to revive from suspend mode ahead of time, completes vernalization；

Fiber differentiation：The sponge for selecting long 20~25cm, wide 15~18cm, 0.5~0.6cm of thickness, it is cleaned with distilled water and is used in combination The alcohol that volume fraction is 75~95% impregnates 30~45 minutes, then is cleaned with distilled water spare；One piece of sponge is rolled into hollow Cylindrical body, bottom fixes one piece of round sponge, and the sponge strip insertion of cylindrical body lower part is equipped in the can of MS culture medium, training The powdered activated carbon for placing 2~3mm thickness on base is supported, 10~15 apertures are inserted to substrate base, in each aperture with glass bar Be inserted perpendicularly into 1~2 plant of young shoot, temperature is 24~26 DEG C, light application time is 12~14 hours, intensity of illumination be 1500~ It is cultivated 20~25 days under the condition of culture of 2000lx；Active carbon help to adjust the bulk density of matrix, total porosity, ventilating slit, The gentle water ratio of water holding hole is easier to evoked callus on the MS culture medium for only adding active carbon, and Callus induction rate reaches 98.0%, callus differentiation is relatively easy to, and callus differentiation rate is up to 99.0%, and the growing state of callus is good, and quality is loose, and color is yellowish green Color is light yellow；

Shoot proliferation culture：The bud section of 2~3 section of tool is taken to access in MS culture medium, Multiplying culture temperature is 22~24 DEG C, when illumination Between be 10~12h, intensity of illumination be 1500~2000Lux, persistently cultivate 25~28 days；By shoot proliferation culture, with less The available a large amount of proliferation seedling of bud primary, cost can be greatly lowered, accelerate the breeding of the white palm, while shoot proliferation Culture can keep stablizing for white palm young shoot primary hereditary；

Tissue culture seedling rooting：The seedling for choosing robust growth, 2~3cm of height, is transferred to the production of inducing adventitious root on root media Raw, culture medium uses 1/2MS culture medium+1/2B5 culture medium for minimal medium, sprinkles the work of 2~3mm thickness in media surface Property carbon powder, controlled at 23~26 DEG C, light application time be 12~14h, intensity of illumination be 2000~2500Lux, training of taking root It can be transplanted after choosing healthy and strong seedling hardening after supporting 15~20 days；The method for culturing seedlings of the white palm is simple and easy, with less bud primary Available a large amount of proliferation seedling, can be greatly lowered cost, accelerate the breeding of the white palm, while the well developed root system of seedling, Root Numerous, diameter are coarse, high survival rate, and after tissue culture and hardening, white palm seedling can adapt to environment quickly, are a kind of The method for culturing seedlings of safe and efficient, cheap environmental protection.

Preferably, adding the carragheen fixed line of 3.5~5.0g/L in all culture mediums, adjusting pH is 5.6~5.8, 121 DEG C of high pressure sterilization 20min.

Preferably, containing 0.2~0.3mg/L (S)-(+) -1- phenyl -2- third in 1/2MS culture medium+1/2B5 culture medium Amine, 0.8~1.2mg/Lt~leucine tert-butyl ester, 0.3~0.5mg/L heteroauxin, 1.2~2.5mg/L methyl α-naphthyl acetate, 3~ 10mg/L zeatin, 15~18mg/L gibberellin；Heteroauxin is degraded with methyl α-naphthyl acetate with air and illumination with photooxidation occurs Timeliness, micro (S)-(+)-amphetamine and t- leucine tert-butyl ester is added simultaneously in the medium can be significantly The degradation rate for delaying heteroauxin, under t- leucine tert-butyl ester atmosphere, (S)-(+)-amphetamine can be with indoles Complex reaction occurs for acetic acid, and generating stable complex compound can prevent illumination from causing to degrade to heteroauxin, and complex compound enters plant It can be used as a kind of deposit form of heteroauxin after in object and be stabilized, dissociate and issue unboiled water explanation to specific environment Heteroauxin is released, its biological effectiveness then occurs；On the one hand heteroauxin, methyl α-naphthyl acetate, zeatin and gibberellin etc. help In sterilizing, can prevent callus from rotting, while the plant cell wall that can also relax, make cell elongation, moreover it is possible to increase RNA and The synthesis of protein can stimulate cambial cell to divide, and stimulate root cell growth, promote xylem, phloem cell differentiation, promote Into the morphogenesis for exceeding injured tissue root of hair, adjusting callus, promotes the generation of adventitious root, grow root system, root item number is more, directly Diameter is coarse, increases white palm survival rate of seedling.

Compared with the prior art, the advantages of the present invention are as follows：

1）When vernalization, under the action of ethanol in proper amount, N- benzylquininium chloride can accelerate its albumen after entering white palm seed Enzyme secretion improves its proteinase activity, while N- benzylquininium chloride can also accelerate the respiratory rate of the white palm, and further Strengthen glycolytic cycle and mitochondria activity, enhances the secretion of soluble sugar, white palm seed is promoted to revive from suspend mode ahead of time；

2）Under t- leucine tert-butyl ester atmosphere, with heteroauxin complex reaction can occur for (S)-(+)-amphetamine, Generating stable complex compound can prevent illumination from causing to degrade to heteroauxin, and complex compound can be used as Yin after entering in plant A kind of deposit form of indolylbutyric acid and be stabilized, dissociating to specific environment issues unboiled water solution and releases heteroauxin, then Its biological effectiveness occurs；

3）The method for culturing seedlings of the white palm is simple and easy, can be significantly with the less available a large amount of proliferation seedling of bud primary Cost is reduced, the breeding of the white palm, while the well developed root system of seedling are accelerated, root Numerous, diameter are coarse, high survival rate, pass through After tissue culture and hardening, white palm seedling can adapt to environment quickly, be a kind of method for culturing seedlings of safe and efficient, cheap environmental protection.

Specific embodiment

The present invention program is described further below by embodiment：

Embodiment 1：

White palm nursery propagation method, specifically includes following steps：

1）Picking shape is full, size is close to intact white palm seed, disinfects 3min, distilled water with 1% liquor natrii hypochloritis It rinses well, is put into vernalization 2h in 0 DEG C of ice bath pot water, contains 10g/L ethyl alcohol, 15mg/LN- benzylquininium chloride in water, To inhale paper suck dry moisture, one layer of filter paper and double gauze are padded in culture dish, are put into seed and are placed in incubator and carry out rudiment；Training Supporting box temperature degree is 24 DEG C, light application time 10h, intensity of illumination 2000Lux, is sprinkled water daily, stops sprinkling after germination；2）Choosing length It is cleaned with distilled water and is used the alcohol that volume fraction is 75% to impregnate 30 points by the sponge of 20cm, width 15cm, thickness 0.5cm Clock, then cleaned with distilled water spare；One piece of sponge is rolled into hollow cylindrical body, one piece of round sponge, cylindrical body are fixed in bottom The sponge strip insertion of lower part places the powdered activated carbon of 2mm thickness, equipped in the can of MS culture medium with glass bar on culture medium 10 apertures are inserted to substrate base, 1 plant of young shoot is inserted perpendicularly into each aperture, temperature is 24 DEG C, light application time is 12 small When, intensity of illumination be 1500Lux condition of culture under cultivate 20 days；3）It takes in the bud section access MS culture medium of 2 sections of tool, proliferation training Supporting temperature is 22 DEG C, light application time 10h, intensity of illumination 1500Lux, is persistently cultivated 25 days；4）Choose robust growth, height The seedling of 2cm, is transferred to the generation of inducing adventitious root on root media, and culture medium is cultivated using 1/2MS culture medium+1/2B5 Base is minimal medium, sprinkles the active carbon powder of 2mm thickness in media surface, and controlled at 23 DEG C, light application time is 12h, intensity of illumination 2000Lux can be transplanted after choosing healthy and strong seedling hardening after culture of rootage 15 days；The method for culturing seedlings of the white palm It is simple and easy, with the less available a large amount of proliferation seedling of bud primary, cost can be greatly lowered, accelerate the numerous of the white palm It educates, while the well developed root system of seedling, root Numerous, diameter are coarse, high survival rate, after tissue culture and hardening, white palm seedling Environment can be adapted to quickly, be a kind of method for culturing seedlings of safe and efficient, cheap environmental protection.

Embodiment 2：

White palm nursery propagation method, specifically includes following steps：

S1：Picking shape is full, size is distilled close to intact white palm seed with 2% liquor natrii hypochloritis disinfection treatment 5min Water is rinsed well, is put into vernalization 3h in 1 DEG C of ice bath pot water, is contained 12g/L ethyl alcohol, 18mg/LN- benzylquininium chlorination in water Object pads one layer of filter paper and double gauze in culture dish, is put into seed and is placed in incubator and sprouted to inhale paper suck dry moisture Bud；Incubator temperature is 26 DEG C, light application time 12h, intensity of illumination 2400Lux, and daily spray water stops spray after germination It spills；Micro N- benzylquininium chloride is added in vernalization aqueous solution and suitable ethyl alcohol can help to vernalization, in appropriate second Under the action of alcohol, N- benzylquininium chloride can stimulate the related gene expression of the white palm after entering white palm seed, accelerate its egg The secretion of white enzyme, improves its proteinase activity, while N- benzylquininium chloride can also accelerate the respiratory rate of the white palm, goes forward side by side one Step strengthens glycolytic cycle and mitochondria activity, enhances the secretion of soluble sugar, white palm seed is promoted to revive from suspend mode ahead of time, Complete vernalization；

S2：The sponge for selecting long 25cm, width 18cm, thickness 0.6cm, it is 95% wine that it is cleaned with distilled water and uses volume fraction Essence is impregnated 45 minutes, then is cleaned with distilled water spare；One piece of sponge is rolled into hollow cylindrical body, one piece of round sea is fixed in bottom Silk floss, the sponge strip insertion of cylindrical body lower part is equipped in the can of MS culture medium, and placement 3mm thickness is powdered activated on culture medium Charcoal inserts 15 apertures to substrate base with glass bar, 2 plants of young shoots is inserted perpendicularly into each aperture, are 26 DEG C, illumination in temperature It is cultivated 25 days under the condition of culture that time is 14 hours, intensity of illumination is 2000lx；Active carbon facilitate adjust matrix bulk density, Total porosity, ventilating slit, the gentle water ratio of water holding hole are easier to callus induction group on the MS culture medium for only adding active carbon It knits, Callus induction rate is up to 98.0%, and callus differentiation is relatively easy to, and callus differentiation rate is up to 99.0%, and the growing state of callus is good, quality Loose, color is yellow green or light yellow；

S3：It takes in the bud section access MS culture medium of 3 sections of tool, Multiplying culture temperature is 24 DEG C, light application time 12h, intensity of illumination For 2000Lux, persistently cultivate 28 days；By shoot proliferation culture, with the less available a large amount of proliferation seedling of bud primary, Cost can be greatly lowered, accelerate the breeding of the white palm, while shoot proliferation culture can keep the stabilization of white palm young shoot primary Heredity；

S4：The seedling for choosing robust growth, height 3cm, is transferred to the generation of inducing adventitious root on root media, culture medium is adopted It is minimal medium with 1/2MS culture medium+1/2B5 culture medium, sprinkles the active carbon powder of 3mm thickness in media surface, controls After temperature is 26 DEG C, light application time 14h, intensity of illumination 2500Lux, culture of rootage 20 days after the healthy and strong seedling hardening of selection i.e. It can transplant；The method for culturing seedlings of the white palm is simple and easy, can be significantly with the less available a large amount of proliferation seedling of bud primary Cost is reduced, the breeding of the white palm, while the well developed root system of seedling are accelerated, root Numerous, diameter are coarse, high survival rate, pass through After tissue culture and hardening, white palm seedling can adapt to environment quickly, be a kind of method for culturing seedlings of safe and efficient, cheap environmental protection.

Embodiment 3：

White palm nursery propagation method, including：Vernalization, Fiber differentiation, shoot proliferation culture, tissue culture seedling rooting, specifically include following Step：

Vernalization：Picking shape is full, size is steamed close to intact white palm seed with 2% liquor natrii hypochloritis disinfection treatment 4min Distilled water is rinsed well, is put into vernalization 2h in 0 DEG C of ice bath pot water, is contained 10g/L ethyl alcohol, 17mg/LN- benzylquininium chlorination in water Object pads one layer of filter paper and double gauze in culture dish, is put into seed and is placed in incubator and sprouted to inhale paper suck dry moisture Bud；Incubator temperature is 25 DEG C, light application time 10h, intensity of illumination 2200Lux, and daily spray water stops spray after germination It spills；Micro N- benzylquininium chloride is added in vernalization aqueous solution and suitable ethyl alcohol can help to vernalization, in appropriate second Under the action of alcohol, N- benzylquininium chloride can stimulate the related gene expression of the white palm after entering white palm seed, accelerate its egg The secretion of white enzyme, improves its proteinase activity, while N- benzylquininium chloride can also accelerate the respiratory rate of the white palm, goes forward side by side one Step strengthens glycolytic cycle and mitochondria activity, enhances the secretion of soluble sugar, white palm seed is promoted to revive from suspend mode ahead of time, Complete vernalization；

Fiber differentiation：It is cleaned with distilled water and is with volume fraction by the sponge for selecting long 25cm, width 15cm, thickness 0.6cm 90% alcohol impregnates 30 minutes, then is cleaned with distilled water spare；One piece of sponge is rolled into hollow cylindrical body, bottom fixes one Block round sponge, the sponge strip insertion of cylindrical body lower part place the powder of 3mm thickness equipped in the can of MS culture medium on culture medium Shaped activated carbon inserts 12 apertures to substrate base with glass bar, 2 plants of young shoots is inserted perpendicularly into each aperture, are 25 in temperature DEG C, cultivate 22 days under the condition of culture that light application time is 12 hours, intensity of illumination is 1800lx；Active carbon helps to adjust matrix Bulk density, total porosity, ventilating slit, the gentle water ratio of water holding hole, only add active carbon MS culture medium on be easier to lure Lead callus, up to 98.0%, callus differentiation is relatively easy to Callus induction rate, and callus differentiation rate is up to 99.0%, the growth feelings of callus Condition is good, and quality is loose, and color is yellow green or light yellow；

Shoot proliferation culture：It takes in the bud section access MS culture medium of 3 sections of tool, Multiplying culture temperature is 23 DEG C, and light application time is 10h, intensity of illumination 1800Lux are persistently cultivated 25 days；It is available big with less bud primary by shoot proliferation culture The proliferation seedling of amount, can be greatly lowered cost, accelerate the breeding of the white palm, while shoot proliferation culture can keep primary white That slaps young shoot stablizes heredity；

Tissue culture seedling rooting：The seedling for choosing robust growth, height 3cm, is transferred to the generation of inducing adventitious root on root media, Culture medium uses 1/2MS culture medium+1/2B5 culture medium for minimal medium, sprinkles the active powdered carbon of 3mm thickness in media surface End, controlled at healthy and strong seedling is chosen after 25 DEG C, light application time 12h, intensity of illumination 2400Lux, culture of rootage 18 days It can be transplanted after hardening；The method for culturing seedlings of the white palm is simple and easy, can with the less available a large amount of proliferation seedling of bud primary Cost is greatly lowered, accelerate the breeding of the white palm, while the well developed root system of seedling, root Numerous, diameter are coarse, survival rate Height, after tissue culture and hardening, white palm seedling can adapt to environment quickly, be a kind of nursery side of safe and efficient, cheap environmental protection Method.

Contain 0.25mg/L (S)-(+)-amphetamine, 1mg/Lt~bright in 1/2MS culture medium+1/2B5 culture medium The propylhomoserin tert-butyl ester, 0.4mg/L heteroauxin, 1.5mg/L methyl α-naphthyl acetate, 8mg/L zeatin, 15mg/L gibberellin；Heteroauxin with Micro (S)-(+) -1- benzene is added photooxidation occurs and timeliness of degrading in methyl α-naphthyl acetate and air and illumination simultaneously in the medium Base -2- propylamine and t- leucine tert-butyl ester can significantly delay the degradation rate of heteroauxin, in t- leucine tert-butyl ester atmosphere Under enclosing, with heteroauxin complex reaction can occur for (S)-(+)-amphetamine, and generating stable complex compound can prevent Illumination causes to degrade to heteroauxin, and complex compound can be used as a kind of deposit form of heteroauxin and steady after entering in plant Fixed to exist, dissociating issues unboiled water solution to specific environment and releases heteroauxin, its biological effectiveness then occurs；Indoles second On the one hand acid, methyl α-naphthyl acetate, zeatin and gibberellin etc. help to sterilize, can prevent callus from rotting, while can be with pine Relaxation plant cell wall, makes cell elongation, moreover it is possible to increase the synthesis of RNA and protein, cambial cell can be stimulated to divide, stimulate root Cell growth promotes xylem, phloem cell differentiation, promotes the morphogenesis for exceeding injured tissue root of hair, adjusting callus, promote The generation for making adventitious root, grows root system, and root item number is more, and diameter is coarse, increases white palm survival rate of seedling.

Routine operation in operation of the present invention step is well known to those skilled in the art, herein without repeating.

Technical solution of the present invention is described in detail in embodiment described above, it should be understood that the above is only For specific embodiments of the present invention, it is not intended to restrict the invention, all any modifications made in spirit of the invention, Supplement or similar fashion substitution etc., should all be included in the protection scope of the present invention.

Claims (10)

1. white palm nursery propagation method, including：Vernalization, Fiber differentiation, shoot proliferation culture, tissue culture seedling rooting, it is characterised in that： The vernalization step is：Picking shape is full, size is close to intact white palm seed, is disinfected with liquor natrii hypochloritis, steams Distilled water is rinsed well, and vernalization in 0~1 DEG C of aqueous solution is put into, to inhale paper suck dry moisture, one layer of filter paper of pad and bilayer in culture dish Gauze, is put into seed and is placed in incubator and carry out rudiment.

2. white palm nursery propagation method according to claim 1, it is characterised in that：Contain in the aqueous solution of the vernalization step There are 10~12g/L ethyl alcohol, 15~18mg/LN- benzylquininium chloride.

3. white palm nursery propagation method according to claim 1, it is characterised in that：In the vernalization step, cultivation temperature It is 24~26 DEG C, light application time is 10~12h, and intensity of illumination is 2000~2400Lux.

4. white palm nursery propagation method according to claim 1, it is characterised in that：The Fiber differentiation step is：It will be extra large Silk floss is cleaned with distilled water and is impregnated with alcohol, then is cleaned with distilled water spare, and one piece of sponge is rolled into hollow cylindrical body, bottom One piece of round sponge is fixed, the sponge strip insertion of cylindrical body lower part places powder equipped in the can of MS culture medium on culture medium Shaped activated carbon is inserted perpendicularly into young shoot with glass bar jack to substrate base in each aperture, cultivates 20~25 days.

5. white palm nursery propagation method according to claim 4, it is characterised in that：In the Fiber differentiation step, culture Temperature is 24~26 DEG C, and light application time is 12~14 hours, and intensity of illumination is 1500~2000lx.

6. white palm nursery propagation method according to claim 1, it is characterised in that：The shoot proliferation incubation step is： The bud section of 2~3 section of tool is taken to access in MS culture medium, Multiplying culture 25~28 days.

7. white palm nursery propagation method according to claim 6, it is characterised in that：In the shoot proliferation incubation step, Cultivation temperature is 22-24 DEG C, and light application time is 10~12h, and intensity of illumination is 1500~2000Lux.

8. white palm nursery propagation method according to claim 1, it is characterised in that：The tissue culture seedling rooting step is：Choosing Robust growth seedling is taken, the generation of inducing adventitious root on root media is transferred to, culture medium uses 1/2MS culture medium+1/2B5 Culture medium is minimal medium, sprinkles active carbon powder in media surface, and healthy and strong seedling is chosen after culture of rootage 15~20 days It can be transplanted after hardening.

9. white palm nursery propagation method according to claim 8, it is characterised in that：It is raw in the tissue culture seedling rooting step Root cultivation temperature is 23~26 DEG C, and light application time is 12~14h, and intensity of illumination is 2000~2500Lux.

10. white palm nursery propagation method according to claim 8, it is characterised in that：In the tissue culture seedling rooting step Contain 0.2~0.3mg/L (S)-(+)-amphetamine, 0.8~1.2mg/Lt in 1/2MS culture medium+1/2B5 culture medium ~leucine tert-butyl ester, 0.3~0.5mg/L heteroauxin, 1.2~2.5mg/L methyl α-naphthyl acetate, 3~10mg/L zeatin, 15~ 18mg/L gibberellin.