CN103392517B - Asexual propagation method of michelia compressa - Google Patents
Asexual propagation method of michelia compressa Download PDFInfo
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- CN103392517B CN103392517B CN201310344978.5A CN201310344978A CN103392517B CN 103392517 B CN103392517 B CN 103392517B CN 201310344978 A CN201310344978 A CN 201310344978A CN 103392517 B CN103392517 B CN 103392517B
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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 a kind of Taiwan is had a smile on one's face.
Background technology
Taiwan with a smile (Michelia compressa), belongs to Magnoliaceae (Magnoliaceae) Michelia (Michelia L.) plant.Evergreen megaphanerophyte, 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 Taiwan is had a smile on one's face is given prominence to, tree-like compact, dryness is comparatively strong, and tree crown is closely coniform, and side shoot is all upward to be stretched, draw close to trunk, dark green and glossiness blade is intensive is born in 1,2 year vimineous top, the tree crown that the formation four seasons are dark green and dense.Spend petite simple and elegant, breezy, give out light fragrance.By expanding numerous popularization, fine tree species are viewed and admired in that Taiwan can become Jiangsu Province and even East China fast-growing with a smile completely, that ornamental value is higher greening.
Grafting refers to the tissue fusion of a plant to the technology on another plant.It is the method for a kind of vegetative propagation plant of horticulture extensive use.Propagation by grafiting views and admires in nursery stock production to solve maternal limited amount or the low main propagation method of cuttage survival rate, produces that the upper stock utilized mainly bred by seedling at present.Grafting has very large meaning to the improvement of plant variety and the preservation of breeding, and gardens have important purposes.The Biological Principles of grafting survival is the surface of anvil fringe bevel, the barrier film of one deck brown is formed by the residue of dead cell, afterwards because of the effect of callus, make the cell division of wound circumference, cambial cell enlivens, break through barrier film and form callus, callus cell breaks up further, inwardly forms new xylem, outwards forms new phloem, the respective xylem vessel of anvil fringe and phloem sieve tube are linked up, makes conducting tissue UNICOM.Be connected with both cork cells along with the cork cell that outside is new is formed, 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 preservation 5 strain is introduced a fine variety in cultivation aspect only Nanjing Botanical Garden Mem. Sun Yat-Sen one place.Maternal quantity is extremely rare, the original producton location of having a smile on one's face, Taiwan is at the greatest differences of weather, soil and other biological condition, Taiwan is caused to introduce a fine variety rear hip number with a smile little, and its seedling to there is reproduction speed slow, breeding cycle is long, and percentage of seedgermination is lower, and seedling such as not easily to survive at the defect, therefore, Taiwan is had a smile on one's face the breeding of plant and is adopted propagation by grafiting more.Liu Xingjian etc., the introduction behave of Taiwan with a smile in Nanjing and propagation by grafiting experiment, disclose is the method that stock grafting Taiwan is had a smile on one's face with 1999 and calendar year 2001 with yulan magnolia respectively, achieve survival rate 27%, the survival rate of 72%, still it is lower to there is survival rate in it, and growing-seedling period is longer, be generally about 2 years, after transplanting, tree vigo(u)r is general, easily endure cold, not easily spend the defects such as winter low temperature, in addition, due to maternal Taiwan, to have a smile on one's face resource quantity extremely few, obtained by tissue culture technique and can increase seedling quantity greatly containing root seedling for grafting, reduce cultivation period, reduce seedling cost, increase economic benefit.
Chinese tulip tree (liriodendrom chinense) belongs to Magnoliaceae, Liriodendron plant, feature is strong adaptability, 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 a large amount of Taiwan fast and effectively 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 the reasonable selection such as stock, grafting method, Taiwan can be made to have a smile on one's face graft survival rate and to reach more than 95%, survive and reach seedling specification fast afterwards, efficiently can obtain Taiwan and have a smile on one's face high quality seedling, greatly meet 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 have a smile on one's face the method for seedling containing root Taiwan, have a smile on one's face re selection for expanding numerous Taiwan in a large number technical support is provided.
Another object of the present invention is to the deficiency overcoming 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 practical easy Taiwan.
By the inventive method, successfully obtain and have a smile on one's face plantlet in vitro containing root Taiwan, proliferation times is 86 times; Wherein adopt subculture multiplication medium, proliferation times is 8 times; Adopt inducing culture, adventitious bud induction frequency is 94%, and adopt root media, rooting rate is 82%, and number of on average taking root is 3.1; Hardening survival rate is to 90%, and can obtain Taiwan efficiently and have a smile on one's face seedling, after stock grafting, survival rate is to more than 95%.
The asexual reproduction method that Taiwan is had a smile on one's face, 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, 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 sterilization: late June, chooses raw top then and, slightly as explant, after tap water is clean, carries out disinfection; Sterilization method is: dip in a little washing powder by its surface brush wash clean with hairbrush, then running water running water is used 30 minutes, the 84 thimerosal shaking tables diluting 50 times to vibrate sterilization 10 minutes, running water is after 3 minutes, with 75% ethanol disinfection 50-60 second on aseptic operating platform, the mercuric chloride pouring 0.1% immediately into is sterilized 10 minutes, finally washs 3 times with sterile water, each vibration 3-5 minute;
(2) preparation of WPM medium is improved: use KNO
3200mg/L replaces the K in former WPM medium
2sO
4, adding concentration of activated carbon is in addition 1000mg/L, and other components are all unchanged.
(3) induction of indefinite bud: get the explant after sterilization in step (1), be cut into the stem section that 0.8cm is long, every stem length of tape one axillalry bud, be inoculated in the induction carrying out indefinite bud in inducing culture, cultivation temperature is 20-26 DEG C, cultivates 12 hours first in the dark, 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 process: utilize high-voltage electrostatic field instrument, carries out static electric field process, and intensity is 10KV/cm, 0.5 hour time;
(5) shoot proliferation: access subculture multiplication medium after being cut by the indefinite bud processed through high-voltage electrostatic field, carry out squamous subculture 20-30 days, cultivation temperature is 20-26 DEG C, light application time is 10-16 hour/day, intensity of illumination is 1600-2000lx, and subculture multiplication medium is for improveing WPM medium and adding 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 after unrooted test-tube plantlet cuts, every strain all has an indefinite bud, culture of rootage 20-30 days is carried out in access root media, cultivation temperature is 20-26 DEG C, 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: be with in step (6) gained Taiwan the blake bottle bottle cap of root aseptic seedling to open with a smile, outdoor cultivation 2-3 days, then move in the matrix adopting carbendazim sterilization, PVC film with holes is used to cover moisturizing above plantlet in vitro, domestication container adopts 50 hole dishes, be placed on ventilating and cooling place and cultivate 25-30 days, cultivate and PVC film was removed in the 15th day, use the sunshade net of index of refraction 75% instead; Described matrix components is red soil and the perlite of weight ratio 1: 1;
All containing agar 6.0g/L, sucrose 20g/L in above-mentioned adventitious bud induction culture base, subculture multiplication medium, root media, and pH value is 5.2.
Adopt the tissue culture propagation that above-mentioned Taiwan is had a smile on one's face, successfully obtain containing root plantlet in vitro, proliferation times is 86 times; Wherein adopt subculture multiplication medium, proliferation times is 8 times; Adopt inducing culture, adventitious bud induction frequency is 94%, and adopt root media, rooting rate is 82%, and number of on average taking root is 3.1; Hardening survival rate, to 90%, can obtain Taiwan efficiently and have a smile on one's face seedling, greatly meet the demand in market.
Step B has a smile on one's face in Taiwan scion pretreatment and is specially:
Get Taiwan that steps A obtains to have a smile on one's face 2 years raw seedlings and germinate branch, upper cut truncation, lower cut are cut sth. askew; Length is about 5cm, retain 1 ~ 2 full bud and prepare scion, scion is soaked in plant growth regulator, described plant growth regulator is (according to weight portion): indolebutyric acid 7 parts, diethyl aminoethyl hexanoate (DA-6) 5 parts, gibberellin 3 parts, KT-301 part, vitamin B1 1 part, ascorbic acid VC 1 part, with the dilution ratio preparation plant growth regulator of 200mg: 1L, then soaks branch bottom 10min with described plant growth regulator, 1%DMSO successively respectively;
Step C grafting method is: grafting time is chosen at March, now the meristematic cell of stock and scion is the most active, vitality is vigorous, be conducive to the healing of calli induction and 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 side incision, then stock is inserted in pretreated scion, after scion is alignd with the formation layer of stock, with Anti-fogging Film, interface is tightly wound around wrapping from bottom to top;
Step D, managing after grafting is:
After grafting completes, attention moisturizing, prevents mechanical damage; Frequent attention observation, maintenance, budder grow to 30 ~ 40cm height time, for not affecting the growth of buddering, timely solution removes the film on interface, but due to stock large, fast growth of buddering, callus is also insecure, that avoids growing tall budders by windbreak, after solution thin film, encloses (can not tension) immediately with straw rope buddering.Budder grow to 60cm time, start to carry out pinching; The early spring sprays 1 0.3% lime sulfur every about 15d, and the primary pest that Taiwan is had a smile on one's face is scale insect.It draws plant juice, and harm young sprout and vein, generally have more the back side of present branch place and leaf.When setting that body branch is overstocked, improper ventilation, easily occur.During control, first to suitably prune overstocked branch off, make tree body ventilation and penetrating light.Manually it can be removed gently with brush when insect pest is lighter; In the nymph incubation period, the 2000 times of liquid sprayings of 40% omethoate emulsifiable solution can be used to be killed.25kg/hm
2, late August Shi Yangfeng composite fertilizer 60kg/hm
2.
The beneficial effect of hinge structure of the present invention is:
The stock that 1 prior art is commonly used and the affinity that Taiwan is had a smile on one's face not strong, the ability producing callus between stock and scion is weak, the effective UNICOM of the conduit of stock and scion, screen casing leads low, new conducting tissue is difficult to be formed, and the substances transport such as moisture and nutrition is not in place, and present inventor is through testing discovery for many years, for the grafting that Taiwan is had a smile on one's face, use Chinese tulip tree stock, affiliation and affinity the best, substantially reduce Chinese tulip tree seedling pipe and support time and cost; Selected stock is the annual Chinese tulip tree stock that the present invention cultivates, director's gesture vigorous period, and Endogenous Hormone Contents in Vitro is high, and stock and scion affinity good, therefore, greatly facilitate graft survival rate.
Respectively with Chinese tulip tree, yulan magnolia, Michelia maudiae, shape accuracy for stock, with a smile, all experiments is the grafting of same grafting work in grafting Taiwan, scion and grafting method all with, investigate different stock and grafting Taiwan had a smile on one's face to the impact of survival rate, growing-seedling period etc.:
Stock of the present invention compares with other stocks
In 2 prior aries, to have a smile on one's face seedling quantity rare in Taiwan, and to take root difficulty due to woody plant, still do not prepare in prior art train in groups containing root seedling:
When WPM medium is cultivated, explant indefinite bud not easily differentiates branch, only quick-fried bud and not extending on explant, and on MS medium, differentiation adventitious buds is leaf bud, to subculture increment not effect.Poor growth in B5, indefinite bud is thin and weak, after being improved by WPM medium, changes NPK ratio, uses KNO
3200mg/L replaces the K in former WPM medium
2sO
4, adding concentration of activated carbon is in addition 1000mg/L, and other components are all unchanged, significantly can promote differentiation and the growth of indefinite bud.
Be below identical Hormone Conditions, different culture media screening unit offshoot program:
The screening of minimal medium
Because of the restriction of damaged, nutrition condition, envirment factor with affect S-type growth curve in 3 adventitious bud inducing growth cycles, but the process of electrostatic field have impact on the permeability of film and the motion of other polar molecule, improve absorption and the transhipment physical capacity of cell, thus the growth of tissue can be promoted, make subculture shortening period of delay, enter exponential phase very soon.Utilize static electric field process, different intensity and ageing can produce different effects, if process electric-field intensity is excessive, or the processing time is long, plant cell can be made to come to harm even dead, and electric-field intensity is too small or the time is too short, treatment effect can be caused again not obvious, present inventor draws through great many of experiments and adopts intensity to be 10KV/cm with a smile to Taiwan, time, halfhour static electric field process significantly can improve upgrowth situation, fresh weight increases obviously, growth rate is 110.7mg/d, greatly facilitate division and the growth of cell, shorten the time of cultured in vitro.
How 4 utilize plant tissue differentiation regeneration plant for plant tissue culture area research, improve the reproductive number 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, present inventor, through a large amount of creative work, 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, adds appropriate active carbon, the oozy poisonous secondary metabolites of absorption plant, makes plant strain growth vigor keep vigorous, plays the effect of containment brownization;
Adopt tissue culture propagation of the present invention, the final rate of increase is 81-86 times; Wherein adopt subculture multiplication medium, proliferation times is 8 times; Adopt inducing culture, adventitious bud induction frequency is 94%, and adopt root media, rooting rate is 82%, and number of on average taking root is 3.1; Hardening survival rate is to 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 promoting or suppress some link in plant life process with lower concentration, making it the wish development to meeting human needs, present inventor is drawn by lot of experiments and to soak through plant growth regulator, Be very effective, described plant growth regulator is according to weight portion: indolebutyric acid 7 parts, diethyl aminoethyl hexanoate (DA-6) 5 parts, gibberellin 3 parts, KT-30 1 part, vitamin B1 1 part, ascorbic acid VC 1 part, with the dilution ratio preparation plant growth regulator of 200mg: 1L, and DMSO is cytoactive component, after 1%DMSO immersion treatment, the time of sprouting to buddering from grafting shortens greatly, single factor experiment draws, than the time decreased 37.3% without any process, plant growth regulator formula of the present invention, it is easy to use, callus activity can be improved, promote callus growth, promoting budders sprouts, improve the chlorophyll content of scion, strengthen photosynthesis, and the auxin rational proportion contained, Coordinated Interaction, there is functions such as promoting root hair cell division, growing of effective raising root hair, obvious shortening seedling-slowing stage, tachyauxesis.
6 the inventive method and other grafting method compare
Embodiment
The asexual reproduction method that Taiwan is had a smile on one's face, is characterized in that comprising the steps: that the cultivation of seedling is had a smile on one's face in A Taiwan, and B has a smile on one's face in Taiwan 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 sterilization: late June, choose raw then top slightly as explant, after tap water is clean, carry out disinfection, sterilization method is: dip in a little washing powder by its surface brush wash clean with hairbrush, then running water running water is used 30 minutes, the 84 thimerosal shaking tables diluting 50 times to vibrate sterilization 10 minutes, running water is after 3 minutes, with 75% ethanol disinfection 50-60 second on aseptic operating platform, the mercuric chloride pouring 0.1% (0.1g mercuric chloride: 100ml water) immediately into is sterilized 10 minutes, finally washs 3 times with sterile water, each vibration 3-5 minute;
(2) preparation of WPM medium is improved: use KNO
3200mg/L replaces the K in former WPM medium
2sO
4, adding concentration of activated carbon is in addition 1000mg/L, and other components are all unchanged.
(3) induction of indefinite bud: get the explant after sterilization in step (1), be cut into the stem section that 0.8cm is long, every stem length of tape one axillalry bud, be inoculated in the induction carrying out indefinite bud in inducing culture, cultivation temperature is 20-26 DEG C, cultivates 12 hours first in the dark, 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 process, utilizes high-voltage electrostatic field instrument, carries out static electric field process, and intensity is 10KV/cm, 0.5 hour time;
(5) shoot proliferation: access subculture multiplication medium after being cut by the indefinite bud processed through high-voltage electrostatic field, carry out squamous subculture 20-30 days, cultivation temperature is 20-26 DEG C, light application time is 10-16 hour/day, intensity of illumination is 1600-2000lx, and subculture multiplication medium is for improveing WPM medium and adding 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 after unrooted test-tube plantlet cuts, every strain all has an indefinite bud, culture of rootage 20-30 days is carried out in access root media, cultivation temperature is 20-26 DEG C, light application time is 10-16 hour/day, intensity of illumination is 1600-2000Lx, and root media is for improveing WPM medium and adding IBA3.0mg/L;
(7) the domestication hardening of aseptic seedling: be with in step (6) gained Taiwan the blake bottle bottle cap of root aseptic seedling to open with a smile,
Outdoor cultivation 2-3 days, then moves in the matrix adopting carbendazim sterilization, uses PVC film with holes to cover moisturizing above plantlet in vitro, domestication container adopts 50 hole dishes, be placed on ventilating and cooling place and cultivate 25-30 days, cultivate and PVC film was removed in the 15th day, use the sunshade net of index of refraction 75% instead; Described matrix components is red soil and the perlite of weight ratio 1: 1;
All containing agar 6.0g/L, sucrose 20g/L in above-mentioned adventitious bud induction culture base, subculture multiplication medium, root media, and pH value is 5.2.
Adopt the tissue culture propagation that above-mentioned Taiwan is had a smile on one's face, successfully obtain containing root plantlet in vitro, proliferation times is 86 times; Wherein adopt subculture multiplication medium, proliferation times is 8 times; Adopt inducing culture, adventitious bud induction frequency is 94%, and adopt root media, rooting rate is 82%, and number of on average taking root is 3.1; Hardening survival rate, to 90%, can obtain Taiwan efficiently and have a smile on one's face seedling, greatly meet the demand in market.
Step B has a smile on one's face in Taiwan scion pretreatment and is specially:
Get Taiwan that steps A obtains to have a smile on one's face 2 years raw seedlings and germinate branch, upper cut truncation, lower cut are cut sth. askew; Length is about 5cm, retain 1 ~ 2 full bud and prepare scion, scion is soaked in plant growth regulator, described plant growth regulator is (according to weight portion): indolebutyric acid 7 parts, diethyl aminoethyl hexanoate (DA-6) 5 parts, gibberellin 3 parts, KT-301 part, vitamin B1 1 part, ascorbic acid VC 1 part, with the dilution ratio preparation plant growth regulator of 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, now the meristematic cell of Chinese tulip tree stock and scion is the most active, vitality is vigorous, be conducive to the healing of calli induction and 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 side incision, then stock is inserted in pretreated scion, after scion is alignd with the formation layer of stock, with Anti-fogging Film, interface is tightly wound around wrapping from bottom to top;
Step D, managing after grafting is:
After grafting completes, attention moisturizing, prevents mechanical damage; Frequent attention observation, maintenance, budder grow to 30 ~ 40cm height time, for not affecting the growth of buddering, timely solution removes the film on interface, but due to stock large, fast growth of buddering, callus is also insecure, that avoids growing tall budders by windbreak, after solution thin film, encloses (can not tension) immediately with straw rope buddering.Budder grow to 60cm time, start to carry out pinching; The early spring sprays 1 0.3% lime sulfur every about 15d, and the primary pest that Taiwan is had a smile on one's face is scale insect.It draws plant juice, and harm young sprout and vein, generally have more the back side of present branch place and leaf.When setting that body branch is overstocked, improper ventilation, easily occur.During control, first to suitably prune overstocked branch off, make tree body ventilation and penetrating light.Manually it can be removed gently with brush when insect pest is lighter; In the nymph incubation period, the 2000 times of liquid sprayings of 40% omethoate emulsifiable solution can be used to be killed.25kg/hm
2, late August Shi Yangfeng composite fertilizer 60kg/hm
2.
Grafting planting percent is had a smile on one's face up to 98% in the Taiwan obtained through the inventive method, and through transplanting land for growing field crops after 2 years, tree vigo(u)r is strong, disease-resistant, can pass the winter safely, efficiently solves Taiwan and has 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 seeds outside case study on implementation implement this method, this will be apparent to those skilled in the art.Therefore, these done without departing from theon the basis of the spirit of the present invention amendments, the expansion of improvement or scope, all belongs to the scope of protection of present invention.
Claims (2)
1. a Taiwan asexual reproduction method of having a smile on one's face, 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, scion pretreatment is had a smile on one's face in Taiwan; Step C, grafting method; And step D, managing after grafting;
Described steps A Taiwan is had a smile on one's face the cultivation of seedling and is specially:
(1) explant is chosen and sterilization: June, chooses the raw top tip then and carries out disinfection as explant;
(2) preparation of WPM medium is improved: use KNO
3200mg/L replaces the K in former WPM medium
2sO
4, the concentration of adding active carbon is in addition 1000mg/L, and other components are all unchanged;
(3) induction of indefinite bud: get the explant after sterilization in step (1), be cut into the stem section that 0.8cm is long, every stem length of tape one axillalry bud, be inoculated in the induction carrying out indefinite bud in adventitious bud induction culture base, cultivation temperature is 20-26 DEG C, cultivates 12 hours first in the dark, then illumination cultivation 20-25 days, described adventitious bud induction culture base is improvement WPM medium prepared by step (2), 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 process: utilize high-voltage electrostatic field instrument, carries out static electric field process to indefinite bud, and intensity is 10KV/cm, 0.5 hour time;
(5) shoot proliferation: by the indefinite bud access subculture multiplication medium processed through high-voltage electrostatic field, carry out squamous subculture 20-30 days and obtain unrooted test-tube plantlet, cultivation temperature is 20-26 DEG C, subculture multiplication medium is improvement WPM medium prepared by step (2), and adds zeatin 0.2mg/L+KT-30 0.2mg/L+IBA0.1mg/L;
(6) culture of rootage: the Taiwan that step (5) obtains is had a smile on one's face carry out culture of rootage 20-30 days in unrooted test-tube plantlet access root media, cultivation temperature is 20-26 DEG C, root media is improvement WPM medium prepared by step (2), and adds IBA3.0mg/L;
Step B has a smile on one's face in Taiwan scion pretreatment and is specially:
The Taiwan that steps A obtains is had a smile on one's face the truncation of seedling branch upper cut, lower cut and cut sth. askew, length is about 5cm, retains 1 ~ 2 full bud, then soaks branch bottom 10min with plant growth regulator, take out branch, then soak branch bottom 10min with 1%DMSO; The preparation method of described plant growth regulator is: get indolebutyric acid 7 parts, diethyl aminoethyl hexanoate 5 parts, gibberellin 3 parts, KT-30 1 part according to weight portion, vitamin B1 1 part, 1 part, ascorbic acid, mix, obtain mixture, with 200mg mixture: the dilution ratio of 1L water adds water, and preparation obtains plant growth regulator;
Step C grafting method is: grafting time is chosen at March, adopts the method for cut-grafting, stock is inserted in pretreated scion, with Anti-fogging Film, interface is tightly wound around wrapping from bottom to top after scion is alignd with the formation layer of stock; Described stock is Chinese tulip tree.
2. method according to claim 1, is characterized in that, in step (1), the concrete steps of sterilization are:
A little washing powder is dipped in by explant surface brush wash clean with hairbrush, then tap water is used 30 minutes, subsequently with sterilization 10 minutes that 84 thimerosal shaking tables of dilution 50 times vibrate, then running water is after 3 minutes, with 75% ethanol disinfection 50-60 second on aseptic operating platform, pour into immediately in the mercuric chloride of 0.1% and sterilize 10 minutes, finally wash 3 times with sterile water, each vibration 3-5 minute.
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|---|---|---|---|---|
| CN103718969B (en) * | 2014-01-28 | 2015-11-18 | 华南农业大学 | A kind of logical cultured in vitro regeneration plant method with a smile of poem beautiful jade |
| CN103858771B (en) * | 2014-04-02 | 2015-09-30 | 山东省农业科学院生物技术研究中心 | A kind of transplanting method of corn gene plantlet in vitro |
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| CN104920084B (en) * | 2015-07-08 | 2017-12-19 | 江苏省林业科学研究院 | A kind of beautiful propagation by grafiting method with a smile |
| 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 |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101049088A (en) * | 2007-05-11 | 2007-10-10 | 中国科学院昆明植物研究所 | Method for breeding plant brand of 'Wanchun Hanxiao |
| KR100889342B1 (en) * | 2007-10-15 | 2009-03-20 | 대한민국 | Propagation method of liriodendron tulipifera using somatic embryogenesis technique |
| CN101695276A (en) * | 2009-10-29 | 2010-04-21 | 中国科学院昆明植物研究所 | Method for cultivating and propagating epipactis mairei |
| CN102217527A (en) * | 2011-05-18 | 2011-10-19 | 中国科学院昆明植物研究所 | Method for breeding michelia variety Yunrui |
| US8227573B2 (en) * | 2005-05-20 | 2012-07-24 | University Of Kentucky Research Foundation | Utility of phylloplanins as antibiotics, selective fungicides and for enhancing microbial resistance in plants |
-
2013
- 2013-08-09 CN CN201310344978.5A patent/CN103392517B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8227573B2 (en) * | 2005-05-20 | 2012-07-24 | University Of Kentucky Research Foundation | Utility of phylloplanins as antibiotics, selective fungicides and for enhancing microbial resistance in plants |
| CN101049088A (en) * | 2007-05-11 | 2007-10-10 | 中国科学院昆明植物研究所 | Method for breeding plant brand of 'Wanchun Hanxiao |
| KR100889342B1 (en) * | 2007-10-15 | 2009-03-20 | 대한민국 | Propagation method of liriodendron tulipifera using somatic embryogenesis technique |
| CN101695276A (en) * | 2009-10-29 | 2010-04-21 | 中国科学院昆明植物研究所 | Method for cultivating and propagating epipactis mairei |
| CN102217527A (en) * | 2011-05-18 | 2011-10-19 | 中国科学院昆明植物研究所 | Method for breeding michelia variety Yunrui |
Non-Patent Citations (2)
| Title |
|---|
| 云南含笑组织培养快繁技术及愈伤组织诱导研究;都婷 等;《云南农业大学学报》;20130703;第28卷(第4期);第530页摘要 * |
| 台湾含笑在南京的引种表现及嫁接繁殖试验;刘兴剑 等;《江苏农业科学》;20081231(第2期);第153页右侧第4段 * |
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