CN103392517A - Asexual propagation method of michelia compressa - Google Patents
Asexual propagation method of michelia compressa Download PDFInfo
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- CN103392517A CN103392517A CN2013103449785A CN201310344978A CN103392517A CN 103392517 A CN103392517 A CN 103392517A CN 2013103449785 A CN2013103449785 A CN 2013103449785A CN 201310344978 A CN201310344978 A CN 201310344978A CN 103392517 A CN103392517 A CN 103392517A
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Abstract
The invention relates to an asexual propagation method of michelia compressa. The asexual propagation method includes the steps that firstly, seedlings of the michelia compressa are manufactured; secondly, pretreatment of a scion of michelia compressa is performed; thirdly, a grafting method is adopted; fourthly, management after grafting is performed. Liriodendron chinense is used as a rootstock, and the cut-grafting method is used for grafting the michelia compressa. After cutting is sequentially soaked into a plant growth regulator and 1%DMSO, the rootstock is inserted, and after the cambium of the scion and the cambium of the rootstock are aligned, an anti-drop film is used for tightly winding and wrapping a port from bottom to top. According to the asexual propagation method of the michelia compressa, a great number of root-included seedlings of the michelia compressa can be obtained, the rate of reproduction is 81-86 times, and after the seedlings are obtained, the seedlings are grafted on the Liriodendron chinense, namely the rootstock, and the survival rate is 95%. After transplantation on a field is achieved for two years, the tree vigor is strong, a tree is resistant to diseases and capable of wintering safely, the problem of vacancy of the market resource of the michelia compressa is effectively solved, and the asexual propagation method of the michelia compressa is wide in application prospect.
Description
Technical field
The invention belongs to plant seedling propagation by grafiting technical field, be specifically related to the asexual reproduction method that have a smile on one's face in a kind of Taiwan.
Background technology
Taiwan is (Michelia compressa) with a smile, belongs to Magnoliaceae (Magnoliaceae) Michelia (Michelia L.) plant.Evergreen megaphanerophyte, the bark taupe, slightly smooth.Ye Quanyuan or wavy edge, keratin, leaf bud and bud have draped over one's shoulders russet fine hair.In Evergreen Broad-leaved Tree Species, the landscape effect that have a smile on one's face in Taiwan is outstanding, tree-like compactness, dryness is stronger, and tree crown is closely coniform, and side shoot is all upward to be stretched, to trunk, draw close, intensive 1,2 year vimineous top, the formation dark green and dense tree crown of the four seasons be born in of dark green and glossiness blade.Spend petite simple and elegantly, breezy, give out light fragrance.By expanding numerous popularization, Taiwan can become greening Jiangsu Province and even East China fast-growing, that ornamental value is higher fully with a smile and view and admire fine tree species.
Grafting refers to the tissue of a plant is fused to technology on another plant.It is the method for a kind of vegetative propagation plant of horticulture extensive use.Propagation by grafiting is to view and admire in nursery stock production to solve maternal limited amount or the low main propagation method of cuttage survival rate, produces at present the upper stock that utilizes and mainly by seedling, breeds.Grafting has very large meaning to the improvement of plant variety and the preservation of breeding, and important purposes is arranged on gardens.The Biological Principles of grafting survival is the surface of anvil fringe bevel, formed the barrier film of one deck brown by the residue of dead cell,, because of the effect of callus, make the cell division of wound circumference afterwards, cambial cell is active, break through barrier film and form callus, callus cell further breaks up, and inwardly forms new xylem, outwards forms new phloem, anvil fringe xylem vessel and phloem sieve tube separately linked up, make conducting tissue UNICOM.Be connected with both cork cells along with the new cork cell in outside forms, final healing becomes a new plant.
Taiwan originates in Taiwan with a smile, is distributed in Taiwan and Japan.Central China finds without wild resource, and cultivation aspect only one place, Nanjing Zhong Shan botanical garden is introduced a fine variety and preserved 5 strains.Maternal quantity is extremely rare, the original producton location that have a smile on one's face in Taiwan is at the greatest differences of weather, soil and other biological condition, cause Taiwan to introduce a fine variety with a smile rear solid amount seldom, and its seedling exists reproduction speed slow, breeding cycle is long, and percentage of seedgermination is lower, and seedling is difficult for the defects such as survival, therefore, Taiwan is had a smile on one's face the breeding of plant and is adopted propagation by grafiting more.Liu Xingjian etc., Taiwan shows and the propagation by grafiting experiment introducing a fine variety of Nanjing with a smile, the method of with 1999 and calendar year 2001, take yulan magnolia, as stock grafting Taiwan, having a smile on one's face is disclosed respectively, obtained survival rate 27%, 72% survival rate, it still exists survival rate lower, and growing-seedling period is longer, be generally about 2 years, after transplanting, tree vigo(u)r is general, easily endure cold, be difficult for spending the defects such as winter low temperature, in addition, to have a smile on one's face resource quantity extremely few due to maternal Taiwan, obtain to contain the root seedling by tissue culture technique and can increase greatly seedling quantity for grafting, reduce cultivation period, reduce seedling cost, increase economic benefit.
Chinese tulip tree (liriodendrom chinense) belongs to Magnoliaceae, the Liriodendron plant, characteristics are strong adaptabilities, growth is fast, resistance, and damage by disease and insect is relatively few, utilizes Chinese tulip tree to do stock grafting Taiwan with a smile, have no report both at home and abroad, its affinity is higher, can obtain fast and effectively a large amount of Taiwan and have a smile on one's face seedling, solves Taiwan and has a smile on one's face the problem of germ plasm resource shortage; By to reasonable selections such as stock, grafting methods, can make Taiwan have a smile on one's face graft survival rate and reach more than 95%, survive the rear seedling specification that reaches fast, can efficiently obtain Taiwan and have a smile on one's face high quality seedling, greatly met the demand in market.
Summary of the invention
The object of the present invention is to provide a kind of a large amount of acquisition to contain root Taiwan and have a smile on one's face the method for seedling, for a large amount of, expand numerous Taiwan and have a smile on one's face the clone plant technical support is provided.
Another object of the present invention is to overcome the deficiency of prior art, provide a kind of simple, convenient, stock injured light, grafting place is easily healed, survival rate is high, efficiency is high, grafting method is had a smile on one's face in the practical easily Taiwan of row.
By the inventive method, successfully obtain to contain root Taiwan group training seedling with a smile, the propagation multiple is 86 times; Wherein adopt the shoot proliferation medium, the propagation multiple is 8 times; Adopt inducing culture, adventitious bud induction frequency is 94%, adopts root media, and rooting rate is 82%, and the number of on average taking root is 3.1; The hardening survival rate reaches 90%, can obtain efficiently Taiwan and have a smile on one's face seedling, and after stock grafting, survival rate reaches more than 95%.
The asexual reproduction method that have a smile on one's face in a kind of Taiwan, is characterized in that comprising the steps: steps A, and the cultivation of seedling is had a smile on one's face in Taiwan; Step B, the scion pretreatment is had a smile on one's face in Taiwan; Step C, grafting method; Step D, managing after grafting;
Wherein steps A Taiwan is had a smile on one's face the cultivation of seedling and is specially:
(1) explant is chosen and is sterilized: late June, and choose and give birth to then top slightly as explant, after running water is rinsed well, carry out disinfection; Sterilization method is: with hairbrush, dip in a little washing powder with its surface brush wash clean, then with running water flowing water, rinsed 30 minutes, diluted on the 84 thimerosal shaking tables of 50 times the vibration sterilization 10 minutes, flowing water rinsed after 3 minutes, on aseptic operating platform with 75% ethanol disinfection 50-60 second, pour immediately 0.1% mercuric chloride sterilization 10 minutes into, with sterile water washing 3 times, 3-5 minute vibrates at every turn finally;
(2) preparation of improvement WPM medium: use KNO
3200mg/L replaces the K in former WPM medium
2SO
4, adding in addition concentration of activated carbon is 1000mg/L, other components are all unchanged.
(3) inducing of indefinite bud: get the explant after sterilization in step (1), be cut into the long stem section of 0.8cm, every stem section is with an axillalry bud, be inoculated in inducing culture and carry out inducing of indefinite bud, cultivation temperature is 20-26 ℃, cultivates at first in the dark 12 hours, then illumination cultivation 20-25 days: light application time is 10-16 hour/day, intensity of illumination is 1600-2000lx, and described adventitious bud induction culture base is improvement WPM medium, and the hormonal readiness of interpolation is:
KT30 1mg/L+ zeatin 0.8mg/L+6-BA0.1mg/L+GA
30.5mg/L;
(4) high-voltage electrostatic field HVEF processes: utilize the high-voltage electrostatic field instrument, carry out electrostatic field and process, intensity is 10KV/cm, 0.5 hour time;
(5) shoot proliferation: will access the shoot proliferation medium after the indefinite bud cutting that process high-voltage electrostatic field, carry out subculture and cultivate 20-30 days, cultivation temperature is 20-26 ℃, light application time is 10-16 hour/day, intensity of illumination is 1600-2000lx, and the shoot proliferation medium is for improvement WPM medium and add zeatin 0.2mg/L+KT-300.2mg/L+IBA0.1mg/L;
(6) culture of rootage: have a smile on one's face after the unrooted test-tube plantlet cuts in the Taiwan that step (5) is obtained, every strain all has an indefinite bud, carry out culture of rootage 20-30 days in the access root media, cultivation temperature is 20-26 ℃, light application time is 10-16 hour/day, intensity of illumination is 1600-2000Lx, and root media improvement WPM medium also adds IBA3.0mg/L;
(7) the domestication hardening of aseptic seedling: step (6) gained Taiwan is opened with the blake bottle bottle cap of root aseptic seedling with a smile, outdoor cultivation 2-3 days, then move in the matrix that adopts the carbendazim sterilization, group training seedling top uses PVC film with holes to cover moisturizing, the domestication container adopts 50 hole dishes, be placed on ventilating and cooling place and cultivate 25-30 days, cultivate and by the 15th day, the PVC film is removed, use the sunshade net of index of refraction 75% instead; Described matrix components is the weight ratio red soil of 1: 1 and perlite;
All contain agar 6.0g/L, sucrose 20g/L in above-mentioned adventitious bud induction culture base, shoot proliferation medium, root media, and the pH value is 5.2.
The tissue culture propagation that adopts above-mentioned Taiwan to have a smile on one's face, successfully obtain to contain root group training seedling, and the propagation multiple is 86 times; Wherein adopt the shoot proliferation medium, the propagation multiple is 8 times; Adopt inducing culture, adventitious bud induction frequency is 94%, adopts root media, and rooting rate is 82%, and the number of on average taking root is 3.1; The hardening survival rate reaches 90%, can obtain efficiently Taiwan and have a smile on one's face seedling, has greatly met the demand in market.
Step B has a smile on one's face in Taiwan the scion pretreatment and is specially:
Get the Taiwan living seedling germinating in 2 years with a smile that steps A obtains, upper cut truncation, lower cut are cut sth. askew; Length is the 5cm left and right, keep 1～2 full bud and prepare scion, scion is soaked in plant growth regulator, described plant growth regulator is (according to weight portion): 7 parts of indolebutyric acids, 5 parts of diethyl aminoethyl hexanoates (DA-6), 3 parts of gibberellin, KT-301 part, 1 part of vitamin B1,1 part of ascorbic acid VC, the dilution ratio preparation plant growth regulator with 200mg: 1L, then soak respectively branch bottom 10min successively with described plant growth regulator, 1%DMSO;
Step C grafting method is: grafting time is chosen at March, this moment, the meristematic cell of stock and scion was the most active, vitality is vigorous, be conducive to the formation of callus and the healing of wound, adopt the method for cut-grafting, stock is cut off in the level and smooth place of bark near the ground, at the long 3～5cm of stock section one side incision, then stock is inserted in pretreated scion, scion is alignd rear with without dripping film, interface tightly being wound around wrapping from bottom to top with the formation layer of stock;
Step D, managing after grafting is:
After grafting is completed, the attention moisturizing, prevent mechanical damage; Often note observation, maintenance, grow to 30～40cm when high buddering, for not affecting the growth of buddering, in time solution is removed the film on interface, but because stock is large, the fast growth of buddering, callus is also insecure, that avoids growing tall budders by windbreak, after separating the thin film, the circle of buddering is lived (can not tension) with the straw rope immediately., buddering while growing to 60cm, start to carry out pinching; Spray 1 0.3% lime sulfur every the 15d left and right early spring, and the primary pest that have a smile on one's face in Taiwan is scale insect.It draws plant juice, endangers young sprout and vein, generally has more the back side of present branch place and leaf., in the situation that tree body branch is overstocked, improper ventilation, easily occur.During control, first to suitably prune overstocked branch off, make tree body ventilation and penetrating light.Can manually with brush, remove gently it when insect pest is lighter; , in the nymph incubation period, can use 2000 times of liquid sprayings of 40% omethoate emulsifiable solution to be killed.25kg/hm
2, late August is executed foreign rich composite fertilizer 60kg/hm
2
The beneficial effect of the relative prior art of the present invention is:
The affinity that have a smile on one's face in the stock that 1 prior art is commonly used and Taiwan is not strong, a little less than producing the ability of callus between stock and scion, the conduit of stock and scion, the effective UNICOM of screen casing lead low, new conducting tissue is difficult to form, and the substances transport such as moisture and nutrition are not in place, and the present application people finds through experiment for many years, the grafting of having a smile on one's face for Taiwan, use the Chinese tulip tree stock, affiliation and affinity are best, have greatly shortened Chinese tulip tree seedling pipe and have supported time and cost; Selected stock is the annual Chinese tulip tree stock that the present invention cultivates, and director's gesture vigorous period, Endogenous Hormone Contents in Vitro is high, and stock and scion affinity good, therefore, greatly promoted graft survival rate.
Respectively take Chinese tulip tree, yulan magnolia, Michelia maudiae, wealthy lobe with a smile as stock, with a smile, all experiments were is the grafting of same grafting work in grafting Taiwan, scion and grafting method all with, investigate different stocks the impact of survival rate, growing-seedling period etc. had a smile on one's face in grafting Taiwan:
The comparison of stock of the present invention and other stocks
In 2 prior aries to have a smile on one's face seedling quantity rare in Taiwan, and due to the woody plant difficulty of taking root, in prior art still not preparation in groups training contain the root seedling:
While cultivating on the WPM medium, the explant indefinite bud is difficult for differentiating branch, quick-fried bud and not extending only on explant, and on the MS medium, differentiation adventitious buds is leaf bud, to not effect of subculture increment.Poor growth in B5, indefinite bud is thin and weak, after the improvement of WPM medium, has changed the NPK ratio, uses KNO
3200mg/L replaces the K in former WPM medium
2SO
4, adding in addition concentration of activated carbon is 1000mg/L, other components are all unchanged, can significantly promote differentiation and the growth of indefinite bud.
Be below identical Hormone Conditions, different culture media screening segmentation scheme:
The screening of minimal medium
In 3 adventitious bud inducing growth cycles because of the restriction of damaged, nutrition condition, envirment factor with affect S-type growth curve, but the processing of electrostatic field has affected the motion of permeability and other polar molecule of film, absorption and the transhipment physical capacity of cell have been improved, thereby can promote the growth of organizing, make subculture shortening period of delay, enter very soon exponential phase.utilize electrostatic field to process, different intensity and ageing can produce different effects, if it is excessive to process electric-field intensity, or the processing time is long, can make plant cell come to harm even dead, and electric-field intensity is too small or the time is too short, can cause again treatment effect not obvious, the present application people's process great many of experiments draws intensity is adopted in Taiwan with a smile is 10KV/cm, halfhour electrostatic field of time is processed and can significantly be improved upgrowth situation, fresh weight increases obviously, growth rate is 110.7mg/d, division and the growth of cell have greatly been promoted, shortened the time of cultured in vitro.
How 4 utilize plant tissue differentiation and regeneration plant for the plant tissue culture area research, improve the breeding quantity of plant, that to study important technology emphasis be also technological difficulties to scientist, and hormone kind and the hormone concentration scope of application, type of culture medium is the most important thing especially, the present application people, through a large amount of creative works, filters out adventitious bud induction culture base, proliferated culture medium and the root media of having a smile on one's face in Taiwan and is respectively:
Improvement WPM medium+KT-30 1mg/L+ zeatin 0.8mg/L+6-BA0.1mg/L+GA
30.5mg/L;
Improvement WPM medium+zeatin 0.2mg/L+KT-300.2mg/L+IBA0.1mg/L;
Improvement WPM medium+IBA3.0mg/L
The WPM medium of improvement, add appropriate active carbon, and the oozy poisonous secondary metabolites of absorption plant makes the plant strain growth vigor keep vigorous, plays the effect of brownization of containment;
Adopt tissue culture propagation of the present invention, the final rate of increase is 81-86 times; Wherein adopt the shoot proliferation medium, the propagation multiple is 8 times; Adopt inducing culture, adventitious bud induction frequency is 94%, adopts root media, and rooting rate is 82%, and the number of on average taking root is 3.1; The hardening survival rate reaches 90%.
5 plant growth regulator are a kind of ectogenic non-nutritive chemical substances, it can conduct to site of action usually in plant corpus, just can promote or suppress some link in the plant life process with lower concentration, make it to the wish development that meets the human needs, the present application people draws through plant growth regulator and soaks by lot of experiments, effect is remarkable, described plant growth regulator is according to weight portion: 7 parts of indolebutyric acids, 5 parts of diethyl aminoethyl hexanoates (DA-6), 3 parts of gibberellin, 1 part of KT-30, 1 part of vitamin B1, 1 part of ascorbic acid VC, dilution ratio preparation plant growth regulator with 200mg: 1L, and DMSO is the cytoactive component, after the 1%DMSO immersion treatment, the time of from grafting, to buddering, sprouting shortens greatly, the single-factor experiment draws, than the time decreased 37.3% without any processing, plant growth regulator formula of the present invention, it is easy to use, can improve callus activity, promote callus growth, promoting budders sprouts, improve the chlorophyll content of scion, strengthen photosynthesis, and the auxin rational proportion that contains, Coordinated Interaction, have the functions such as the root hair cell of promotion division, effectively improve growing of root hair, obviously shorten seedling-slowing stage, tachyauxesis.
6 the inventive method and other grafting method are relatively
Embodiment
The asexual reproduction method that have a smile on one's face in a kind of Taiwan, is characterized in that comprising the steps: that A Taiwan has a smile on one's face the cultivation of seedling, and B has a smile on one's face in Taiwan the scion pretreatment, C grafting method, D, managing after grafting;
Wherein A Taiwan is had a smile on one's face the cultivation of seedling and is specially:
(1) explant is chosen and is sterilized: late June, choose and give birth to then top slightly as explant, after running water is rinsed well, carry out disinfection, sterilization method is: with hairbrush, dip in a little washing powder with its surface brush wash clean, then with running water flowing water, rinsed 30 minutes, diluted on the 84 thimerosal shaking tables of 50 times the vibration sterilization 10 minutes, flowing water rinsed after 3 minutes, on aseptic operating platform with 75% ethanol disinfection 50-60 second, (0.1g mercuric chloride: mercuric chloride 100ml water) was sterilized 10 minutes, and with sterile water washing 3 times, 3-5 minute vibrates at every turn finally to pour immediately 0.1% into;
(2) preparation of improvement WPM medium: use KNO
3200mg/L replaces the K in former WPM medium
2SO
4, adding in addition concentration of activated carbon is 1000mg/L, other components are all unchanged.
(3) inducing of indefinite bud: get the explant after sterilization in step (1), be cut into the long stem section of 0.8cm, every stem section is with an axillalry bud, be inoculated in inducing culture and carry out inducing of indefinite bud, cultivation temperature is 20-26 ℃, cultivates at first in the dark 12 hours, then illumination cultivation 20-25 days: light application time is 10-16 hour/day, intensity of illumination is 1600-2000lx, and described inducing culture is improvement WPM medium, and the hormonal readiness of interpolation is:
KT-30 1mg/L+ zeatin 0.8mg/L+6-BA0.1mg/L+GA
30.5mg/L;
(4) high-voltage electrostatic field HVEF processes, and utilizes the high-voltage electrostatic field instrument, carries out electrostatic field and processes, and intensity is 10KV/cm, 0.5 hour time;
(5) shoot proliferation: will access the shoot proliferation medium after the indefinite bud cutting that process high-voltage electrostatic field, carry out subculture and cultivate 20-30 days, cultivation temperature is 20-26 ℃, light application time is 10-16 hour/day, intensity of illumination is 1600-2000lx, and the shoot proliferation medium is for improvement WPM medium and add zeatin 0.2mg/L+KT-300.2mg/L+IBA0.1mg/L;
(6) culture of rootage: have a smile on one's face after the unrooted test-tube plantlet cuts in the Taiwan that step (5) is obtained, every strain all has an indefinite bud, carry out culture of rootage 20-30 days in the access root media, cultivation temperature is 20-26 ℃, light application time is 10-16 hour/day, intensity of illumination is 1600-2000Lx, and root media is for improvement WPM medium and add IBA3.0mg/L;
(7) the domestication hardening of aseptic seedling: step (6) gained Taiwan is opened with the blake bottle bottle cap of root aseptic seedling with a smile,
Outdoor cultivation 2-3 days, then move in the matrix that adopts the carbendazim sterilization, and group training seedling top uses PVC film with holes to cover moisturizing, the domestication container adopts 50 hole dishes, be placed on ventilating and cooling place and cultivate 25-30 days, cultivate and by the 15th day, the PVC film is removed, use the sunshade net of index of refraction 75% instead; Described matrix components is the weight ratio red soil of 1: 1 and perlite;
All contain agar 6.0g/L, sucrose 20g/L in above-mentioned adventitious bud induction culture base, shoot proliferation medium, root media, and the pH value is 5.2.
The tissue culture propagation that adopts above-mentioned Taiwan to have a smile on one's face, successfully obtain to contain root group training seedling, and the propagation multiple is 86 times; Wherein adopt the shoot proliferation medium, the propagation multiple is 8 times; Adopt inducing culture, adventitious bud induction frequency is 94%, adopts root media, and rooting rate is 82%, and the number of on average taking root is 3.1; The hardening survival rate reaches 90%, can obtain efficiently Taiwan and have a smile on one's face seedling, has greatly met the demand in market.
Step B has a smile on one's face in Taiwan the scion pretreatment and is specially:
Get the Taiwan living seedling germinating in 2 years with a smile that steps A obtains, upper cut truncation, lower cut are cut sth. askew; Length is the 5cm left and right, keep 1～2 full bud and prepare scion, scion is soaked in plant growth regulator, described plant growth regulator is (according to weight portion): 7 parts of indolebutyric acids, 5 parts of diethyl aminoethyl hexanoates (DA-6), 3 parts of gibberellin, KT-301 part, 1 part of vitamin B1,1 part of ascorbic acid VC, the dilution ratio preparation plant growth regulator with 200mg: 1L, then soak branch bottom 10min successively with described plant growth regulator, 1% (V/V) DMSO;
Step C grafting method is: grafting time is chosen at March, this moment, the meristematic cell of Chinese tulip tree stock and scion was the most active, vitality is vigorous, be conducive to the formation of callus and the healing of wound, adopt the method for cut-grafting, stock is cut off in the level and smooth place of bark near the ground, at the long 3～5cm of stock section one side incision, then stock is inserted in pretreated scion, scion is alignd rear with without dripping film, interface tightly being wound around wrapping from bottom to top with the formation layer of stock;
Step D, managing after grafting is:
After grafting is completed, the attention moisturizing, prevent mechanical damage; Often note observation, maintenance, grow to 30～40cm when high buddering, for not affecting the growth of buddering, in time solution is removed the film on interface, but because stock is large, the fast growth of buddering, callus is also insecure, that avoids growing tall budders by windbreak, after separating the thin film, the circle of buddering is lived (can not tension) with the straw rope immediately., buddering while growing to 60cm, start to carry out pinching; Spray 1 0.3% lime sulfur every the 15d left and right early spring, and the primary pest that have a smile on one's face in Taiwan is scale insect.It draws plant juice, endangers young sprout and vein, generally has more the back side of present branch place and leaf., in the situation that tree body branch is overstocked, improper ventilation, easily occur.During control, first to suitably prune overstocked branch off, make tree body ventilation and penetrating light.Can manually with brush, remove gently it when insect pest is lighter; , in the nymph incubation period, can use 2000 times of liquid sprayings of 40% omethoate emulsifiable solution to be killed.25kg/hm
2, late August is executed foreign rich composite fertilizer 60kg/hm
2
Have a smile on one's face the grafting planting percent up to 98% through the Taiwan that the inventive method obtains, and through transplanting land for growing field crops after 2 years, tree vigo(u)r is strong, and is disease-resistant, can pass the winter safely, efficiently solves Taiwan and have a smile on one's face the vacancy of market resource, have broad application prospects.
Although, above the present invention is described in detail with a general description of the specific embodiments, but on basis of the present invention, can make some modifications or improvements it, or the enforcement of the seeds outside case study on implementation this method, this will be apparent to those skilled in the art.Therefore, these that make are without departing from theon the basis of the spirit of the present invention revised, and the expansion of improvement or scope, all belong to the scope of protection of present invention.
Claims (3)
1. the asexual reproduction method that have a smile on one's face in Taiwan, is characterized in that described method comprises the steps: steps A, and the cultivation of seedling is had a smile on one's face in Taiwan; Step B, the scion pretreatment is had a smile on one's face in Taiwan; Step C, grafting method; And step D, managing after grafting.
2. the method for claim 1, is characterized in that, steps A Taiwan is had a smile on one's face the cultivation of seedling and is specially:
(1) explant is chosen and is sterilized: in June, choose living top then and slightly as explant, carry out disinfection;
(2) preparation of improvement WPM medium: use KNO
3200mg/L replaces the K in former WPM medium
2SO
4, the concentration of adding in addition active carbon is 1000mg/L, other components are all unchanged;
(3) inducing of indefinite bud: get the explant after sterilization in step (1), be cut into the long stem section of 0.8cm, every stem section is with an axillalry bud, be inoculated in the adventitious bud induction culture base and carry out inducing of indefinite bud, cultivation temperature is 20-26 ℃, cultivates at first in the dark 12 hours, then illumination cultivation 20-25 days, described adventitious bud induction culture base is the improvement WPM medium of step (2) preparation, and the hormonal readiness of interpolation is:
KT-30 1mg/L+ zeatin 0.8mg/L+6-BA0.1mg/L+GA
30.5mg/L;
(4) high-voltage electrostatic field HVEF processes: utilize the high-voltage electrostatic field instrument, indefinite bud is carried out electrostatic field process, intensity is 10KV/cm, 0.5 hour time;
(5) shoot proliferation: the indefinite bud access shoot proliferation medium that will process through high-voltage electrostatic field, carry out the subculture cultivation and obtained the unrooted test-tube plantlet in 20-30 days, cultivation temperature is 20-26 ℃, the shoot proliferation medium is the improvement WPM medium of step (2) preparation, and adds zeatin 0.2mg/L+KT-300.2mg/L+IBA0.1mg/L;
(6) culture of rootage: the Taiwan that step (5) obtains is had a smile on one's face to carry out culture of rootage 20-30 days in unrooted test-tube plantlet access root media, cultivation temperature is 20-26 ℃, root media is the improvement WPM medium of step (2) preparation, and adds IBA3.0mg/L;
Step B has a smile on one's face in Taiwan the scion pretreatment and is specially:
The Taiwan that steps A is obtained is had a smile on one's face the truncation of seedling branch upper cut, lower cut and is cut sth. askew, length is the 5cm left and right, keeps 1～2 full bud, then with plant growth regulator, soaks branch bottom 10min, take out branch, then with 1%DMSO, soak branch bottom 10min; The preparation method of described plant growth regulator is: according to weight portion, get 7 parts of indolebutyric acids, 5 parts of diethyl aminoethyl hexanoates, 3 parts of gibberellin, 1 part of KT-30,1 part of vitamin B1,1 part, ascorbic acid, mix, obtain mixture, with the 200mg mixture: the dilution ratio of 1L water is added water, and preparation obtains plant growth regulator;
Step C grafting method is: grafting time is chosen at March, adopts the method for cut-grafting, and stock is inserted in pretreated scion, scion is alignd rear with without dripping film, interface tightly being wound around wrapping from bottom to top with the formation layer of stock; Described stock is Chinese tulip tree.
3. method according to claim 2, is characterized in that, in step (1), the concrete steps of sterilization are:
Dip in a little washing powder with explant surface brush wash clean with hairbrush, then with running water, rinsed 30 minutes, subsequently with vibration sterilization on 84 thimerosal shaking tables of 50 times of dilutions 10 minutes, then flowing water rinsed after 3 minutes, on aseptic operating platform with 75% ethanol disinfection 50-60 second, poured immediately in 0.1% mercuric chloride sterilization into 10 minutes, with sterile water washing 3 times, 3-5 minute vibrates at every turn finally.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310344978.5A CN103392517B (en) | 2013-08-09 | 2013-08-09 | Asexual propagation method of michelia compressa |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310344978.5A CN103392517B (en) | 2013-08-09 | 2013-08-09 | Asexual propagation method of michelia compressa |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103392517A true CN103392517A (en) | 2013-11-20 |
| CN103392517B CN103392517B (en) | 2015-05-13 |
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| CN103718969A (en) * | 2014-01-28 | 2014-04-16 | 华南农业大学 | Regenerated plant in-vitro culture method for Sirindhorn michelia figo |
| CN103858771A (en) * | 2014-04-02 | 2014-06-18 | 山东省农业科学院生物技术研究中心 | Maize transgenic tissue culture seedling transplanting method |
| CN104126492A (en) * | 2014-08-06 | 2014-11-05 | 江苏省中国科学院植物研究所 | In-vitro propagation method of taxodium hybrids 'zhongshansha136' |
| CN104396656A (en) * | 2014-09-23 | 2015-03-11 | 常州市金土地农牧科技服务有限公司 | Yield-increasing method of pecans |
| CN104904499A (en) * | 2014-05-09 | 2015-09-16 | 常州市金土地农牧科技服务有限公司 | Method for grafting caryaillinoinensis |
| CN104920084A (en) * | 2015-07-08 | 2015-09-23 | 江苏省林业科学研究院 | Grafting propagation method for jinxiu michelia figo |
| CN105815169A (en) * | 2015-12-03 | 2016-08-03 | 胡晓敏 | Method for grafting and breeding michelia shiluensis |
| CN106879459A (en) * | 2017-03-01 | 2017-06-23 | 周口师范学院 | A kind of plant induces the breeding method of breeding objective Mutations In Plants |
| CN107148860A (en) * | 2017-05-08 | 2017-09-12 | 茂名市绿凰农业发展有限公司 | It is a kind of that the engrafting method of the high-quality and efficient safety in production of crop is promoted based on high-voltage electrostatic field |
| CN109548505A (en) * | 2019-01-17 | 2019-04-02 | 曹春花 | The grafting and breeding technique of Magnolia wufengensis |
| CN109566130A (en) * | 2019-01-25 | 2019-04-05 | 曹春花 | A kind of method of scale fast breeding Magnolia wufengensis |
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| CN104904499A (en) * | 2014-05-09 | 2015-09-16 | 常州市金土地农牧科技服务有限公司 | Method for grafting caryaillinoinensis |
| CN104126492A (en) * | 2014-08-06 | 2014-11-05 | 江苏省中国科学院植物研究所 | In-vitro propagation method of taxodium hybrids 'zhongshansha136' |
| CN104126492B (en) * | 2014-08-06 | 2016-04-13 | 江苏省中国科学院植物研究所 | The in-vitro propagation method of a kind of middle mountain China fir kind 136 |
| CN104396656A (en) * | 2014-09-23 | 2015-03-11 | 常州市金土地农牧科技服务有限公司 | Yield-increasing method of pecans |
| CN104920084A (en) * | 2015-07-08 | 2015-09-23 | 江苏省林业科学研究院 | Grafting propagation method for jinxiu michelia figo |
| CN105815169A (en) * | 2015-12-03 | 2016-08-03 | 胡晓敏 | Method for grafting and breeding michelia shiluensis |
| CN106879459A (en) * | 2017-03-01 | 2017-06-23 | 周口师范学院 | A kind of plant induces the breeding method of breeding objective Mutations In Plants |
| CN107148860A (en) * | 2017-05-08 | 2017-09-12 | 茂名市绿凰农业发展有限公司 | It is a kind of that the engrafting method of the high-quality and efficient safety in production of crop is promoted based on high-voltage electrostatic field |
| CN109548505A (en) * | 2019-01-17 | 2019-04-02 | 曹春花 | The grafting and breeding technique of Magnolia wufengensis |
| CN109566130A (en) * | 2019-01-25 | 2019-04-05 | 曹春花 | A kind of method of scale fast breeding Magnolia wufengensis |
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