CN105519434B - A kind of tissue-cultured seedling micro-grafting method of Japanese maple orange dream - Google Patents
A kind of tissue-cultured seedling micro-grafting method of Japanese maple orange dream Download PDFInfo
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- CN105519434B CN105519434B CN201511023601.5A CN201511023601A CN105519434B CN 105519434 B CN105519434 B CN 105519434B CN 201511023601 A CN201511023601 A CN 201511023601A CN 105519434 B CN105519434 B CN 105519434B
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G2/00—Vegetative propagation
- A01G2/30—Grafting
Abstract
The present invention discloses a kind of tissue-cultured seedling micro-grafting method of Japanese maple orange dream, including (1) sterile stock is cultivated：Fructus Aceris elegantulis band bud branch section is cut, tissue cultures is carried out, obtains Fructus Aceris elegantulis aseptic seedling, the beveling of Fructus Aceris elegantulis aseptic seedling is blocked, obtains sterile stock；(2) sterile scion cultivated：Japanese maple orange dream band bud branch section is cut, tissue cultures is carried out, obtains Japanese maple orange dream aseptic seedling, the beveling of Japanese maple orange dream aseptic seedling is blocked, obtains sterile scion；(3) tissue-cultured seedling micrografting：The oblique section of sterile stock is alignd with the oblique section of sterile scion, and it is fixed, micrografting seedling is obtained, the grafting is inoculated on culture medium and cultivated；(4) acclimatization and transplantses：Micrografting seedling obtained by step (3) is transferred to greenhouse hardening at initial stage, then is transferred to hardening in seedbed, spraying treatment and sterilization processing, hardening can transplant after terminating；Methods described can improve grafting efficiency, improve reproductive number, survival rate is high, can carry out cultivating small dimension Japan maple orange dream seedling on a large scale.
Description
Technical field
The present invention relates to propagation by grafiting technical field, and in particular to a kind of tissue-cultured seedling micrografting side of Japanese maple orange dream
Method.
Background technology
The ornamental value of Japanese maple orange dream (Acer palmatum " Orange Dream ") is high, and kind is unique, spring leaf
Color is greenish-yellow, and more by solar radiation, leaf color is more changed into yellow, and limb is general red, gradually to be orange, is changed into as smart as a new pin to summer
Orange, vein yellow, autumn for golden yellow, the leaf color change of three seasons is obvious, and all the time burnt phenomenon under burning sun, very joyous by market
Meet.
At present, Japanese maple orange dream is subject to seasonal restrictions mainly by traditional grafting method, and reproductive number is few, and survival rate is low,
The defects of cost of labor is higher, and minimum 1cm of stock specification or so, it is impossible to produce small dimension grafting, it is impossible to meet micro- basin
Planting needs.
The content of the invention
In view of this, the application provides a kind of tissue-cultured seedling micro-grafting method of Japanese maple orange dream, and methods described can improve
The efficiency of grafting, reproductive number being improved, environment is controllable, and survival rate is high, and grafting cultivation period is short, can not be subject to seasonal restrictions,
Overcome traditional grafting method to be subject to seasonal restrictions, the defects of reproductive number is few, and survival rate is low, and cost of labor is higher, can carry out big
The industrialization nursery and deep processing of scale.
To solve above technical problem, technical scheme provided by the invention, which is that a kind of tissue-cultured seedling of Japanese maple orange dream is micro-, transfers
Method is connect, the described method comprises the following steps：
(1) sterile stock is cultivated：Fructus Aceris elegantulis band bud branch section is cut, tissue cultures is carried out, obtains Fructus Aceris elegantulis aseptic seedling, will be beautiful
The beveling of maple aseptic seedling is blocked, and obtains sterile stock；
(2) sterile scion cultivated：Japanese maple orange dream band bud branch section is cut, tissue cultures is carried out, obtains Japanese maple orange dream
Aseptic seedling, the beveling of Japanese maple orange dream aseptic seedling is blocked, obtains sterile scion；
(3) tissue-cultured seedling micrografting：By the oblique section of the sterile stock described in step (1) and step (2) the sterile scion
Oblique section alignment, it is and fixed, obtain micrografting seedling, the grafting be inoculated in the week of culture 4-5, the trainings on culture medium
Support base composition be：MS minimal mediums, supplement 0.25-1.50mg/L 6-BA, 0.02-0.10mg/L NAA, 5-30g/L
Sucrose；
(4) acclimatization and transplantses：Micrografting seedling obtained by step (3) is transferred to greenhouse hardening at initial stage 5-8 days, then is transferred in seedbed
In the week of hardening 4-5, in the seedbed during hardening, carrying out spraying treatment and sterilization processing, hardening can transplant after terminating.
Preferably, in the tissue-cultured seedling micrografting, the condition that micrografting seedling is cultivated in the medium is：Temperature is 23-27
DEG C, intensity of illumination is 2300-2600Lx, light application time 12h/d.
Preferably, in the tissue-cultured seedling micrografting, the composition of the culture medium is：MS minimal mediums, supplement 0.5-
1.25mg/L 6-BA, 0.03-0.07mg/L NAA, 10-25g/L sucrose.
It is more highly preferred to, in the tissue-cultured seedling micrografting, the composition of the culture medium is：MS minimal mediums, supplement
1.0mg/L 6-BA, 0.05mg/L NAA, 20g/L sucrose.
Preferably, in the tissue-cultured seedling micrografting, 3-7g/L agar, the pH of the agar are also supplemented in the culture medium
It is worth for 6.5.
Preferably, the stem of the sterile stock is slightly more than 1.5mm, and the stem of the sterile scion is slightly more than 1mm.
Preferably, in the acclimatization and transplantses, the matrix in the seedbed is that volume ratio is 2：The perlite and humic of (2-4)
The mixture of matter.
Preferably, in the acclimatization and transplantses, the method for the spraying treatment is：During fine day, sprayed at interval of 10-15min
3—4s；Rainy day or it is cloudy when, sprayed 3-4s at interval of 18-25min, time of spraying treatment is 6-12 days.
Preferably, in the acclimatization and transplantses, the bactericidal treatments are：It is comprehensive using microbicide solution every 5-8 days
Spray grafting.
It is more highly preferred to, the microbicide solution is selected from 70% thiophanate methyl, 800 times of solution, more than 50% for any one
800 times of solution of bacterium spirit, 75% Bravo, 800 times of solution.
Wherein, during the sterile stock is cultivated, the method for the tissue cultures is：By Fructus Aceris elegantulis band bud branch section sterilization
After be seeded on axillary bud deriving culture medium and cultivate；The axillary bud of sprouting is cut, is seeded on inducing clumping bud culture medium and cultivates；Will
The Multiple Buds differentiated are cut into individual plant, are inoculated with root media culture, produce Fructus Aceris elegantulis aseptic seedling.
In the sterile scion cultivated, the method for the tissue cultures is：The band bud branch section sterilization of Japanese maple orange dream is gone out
It is seeded on axillary bud deriving culture medium and cultivates after bacterium；The axillary bud of sprouting is cut, is seeded on inducing clumping bud culture medium and cultivates；
The Multiple Buds differentiated are cut into individual plant, root media culture is inoculated with, produces Japanese maple orange dream aseptic seedling.
MS minimal mediums described herein have higher inorganic salt concentration, can ensure the ore deposit needed for tissue growth
Matter nutrition, moreover it is possible to accelerate the growth of callus, be more stable ionic equilibrium solution, its nitrate content is high, its nutrient
Quantity and ratio it is suitable, the nutrition and physiological requirements of plant cell can be met, thus the scope of application is wider, most plants group
Knit the minimal medium that the quick breeding of culture uses it as culture medium.
The 6-BA is 6- benzyl aminoadenines, is a kind of auxin, and its main function is the formation for promoting bud,
Can also evoked callus occur, promote cell division, promote the differentiation of undifferentiated tissue, promote the product of biological substance in vivo
It is tired, promote lateral bud, prevent aging, be the basic element of cell division in plant tissue and cell culture.
The NAA is methyl α-naphthyl acetate, is a kind of auxin, is used when plant is using cuttage breeding, it is also possible to
In Plant Tissue Breeding, cell division can be promoted with expanding, induced synthesis adventitious root increase fruit setting, shedding is prevented, change female, male
Flower ratio etc., tender epidermis that can be through blade, branch, seed is entered in plant, with nutrition stream transporting to complete stool.
Compared with prior art, its detailed description is as follows by the application：Technical scheme provides a kind of Japanese maple orange
Dream tissue-cultured seedling micro-grafting method, including sterile stock cultivation, sterile scion cultivated, tissue-cultured seedling micrografting, acclimatization and transplantses
Step, by tissue-cultured seedling micrografting step, the screening of culture medium, optimal nutrient media components and proportioning being obtained, before
To state auxin and match the culture medium to be formed, formula is simple, and culture medium cost is low, can accelerate the healing of micrografting wound,
Ensure the survival rate of micrografting more than 90%, thus improve the efficiency of grafting, reproductive number is improved, because its environment can
Control, survival rate is high, and grafting cultivation period is short, can not be subject to seasonal restrictions, therefore overcomes traditional grafting method and limited by season
System, the defects of reproductive number is few, and survival rate is low, and cost of labor is higher, can the smaller grafting of production specification be used for micro- potted landscape, micro-
It is potted plant, solve tissue culture and take root difficult problem, large-scale industrialization nursery and deep processing can be carried out, there is good application prospect.
Embodiment
In order that those skilled in the art more fully understands technical scheme, with reference to specific embodiment pair
The present invention is described in further detail.
Embodiment 1
The tissue-cultured seedling micro-grafting method of Japanese maple orange dream described in the present embodiment, comprises the following steps：
(1) sterile stock is cultivated：Fructus Aceris elegantulis band bud branch section is cut, is seeded on axillary bud deriving culture medium and trains after sterilization
Support；The axillary bud of sprouting is cut, is seeded on inducing clumping bud culture medium and cultivates；The Multiple Buds differentiated are cut into individual plant, connect
Kind root media culture, obtains Fructus Aceris elegantulis aseptic seedling, and the beveling of Fructus Aceris elegantulis aseptic seedling is blocked, obtains sterile stock, stem is slightly 1.5mm
More than；
(2) sterile scion cultivated：Japanese maple orange dream band bud branch section is cut, axillary bud deriving culture is seeded to after sterilization
Cultivated on base；The axillary bud of sprouting is cut, is seeded on inducing clumping bud culture medium and cultivates；The Multiple Buds differentiated are cut into list
Strain, root media culture is inoculated with, obtains Japanese maple orange dream aseptic seedling, the beveling of Japanese maple orange dream aseptic seedling is blocked, obtains nothing
Bacterium scion, stem are slightly more than 1mm；
(3) tissue-cultured seedling micrografting：By the oblique section of the sterile stock described in step (1) and step (2) the sterile scion
Oblique section alignment, and fixed using sterile emulsion tube, obtain micrografting seedling, the grafting is inoculated on culture medium and cultivated
4-5 weeks, cultivation temperature are 23-27 DEG C, and intensity of illumination is 2300-2600Lx, light application time 12h/d.The culture medium
Form and be：MS minimal mediums, supplement 1.0mg/L 6-BA, 0.05mg/L NAA, 20g/L sucrose, wherein 5g/L agar, agar
PH value be 6.5；
(4) acclimatization and transplantses：Micrografting seedling obtained by step (3) is transferred to greenhouse hardening at initial stage 7 days, then is transferred to hardening in seedbed
In 4-5 weeks, seedbed was carried out disinfection in 7 days in advance, the matrix in seedbed is that volume ratio is 2：3 perlite and the mixture of humus,
In the seedbed during hardening, carrying out spraying treatment and sterilization processing, hardening can transplant after terminating, wherein at the spraying
The method of reason is：During fine day, at interval of 10min sprayings 3s；Rainy day or it is cloudy when, at interval of 20min spray 3s, spraying treatment
Time is 10 days, and the bactericidal treatments are：Grafting, the bactericide were sprayed comprehensively using microbicide solution every 7 days
Solution is 70% thiophanate methyl, 800 times of solution.
Embodiment 2
The tissue-cultured seedling micro-grafting method of Japanese maple orange dream described in the present embodiment, the difference with embodiment 1 are：
In step (3) tissue-cultured seedling micrografting, the composition of the culture medium is：MS minimal mediums, supplement 1.0mg/L 6-
BA, 0.05mg/L NAA, 20g/L sucrose, 4g/L agar.
Embodiment 3
The tissue-cultured seedling micro-grafting method of Japanese maple orange dream described in the present embodiment, the difference with embodiment 1 are：
In step (3) tissue-cultured seedling micrografting, the composition of the culture medium is：MS minimal mediums, supplement 0.5mg/L 6-
BA, 0.03mg/L NAA, 10g/L sucrose, 4g/L agar.
Embodiment 4
The tissue-cultured seedling micro-grafting method of Japanese maple orange dream described in the present embodiment, the difference with embodiment 1 are：
In step (3) tissue-cultured seedling micrografting, the composition of the culture medium is：MS minimal mediums, supplement 1.25mg/L 6-
BA, 0.07mg/L NAA, 25g/L sucrose, 5g/L agar.
Embodiment 5
The tissue-cultured seedling micro-grafting method of Japanese maple orange dream described in the present embodiment, the difference with embodiment 1 are：
In step (3) tissue-cultured seedling micrografting, the composition of the culture medium is：MS minimal mediums, supplement 0.25mg/L 6-
BA, 0.02mg/L NAA, 5g/L sucrose, 4g/L agar.
Embodiment 6
The tissue-cultured seedling micro-grafting method of Japanese maple orange dream described in the present embodiment, the difference with embodiment 1 are：
In step (3) tissue-cultured seedling micrografting, the composition of the culture medium is：MS minimal mediums, supplement 1.50mg/L 6-
BA, 0.10mg/L NAA, 30g/L sucrose, 5g/L agar.
Embodiment 7
The tissue-cultured seedling micro-grafting method of Japanese maple orange dream described in the present embodiment, the difference with embodiment 1 are：
In step (3) tissue-cultured seedling micrografting, MS minimal mediums, supplement 0.1mg/L 6-BA, 0.08mg/L NAA, 20g/
L sucrose, 5g/L agar.
Embodiment 8
The tissue-cultured seedling micro-grafting method of Japanese maple orange dream described in the present embodiment, the difference with embodiment 1 are：
In step (3) tissue-cultured seedling micrografting, MS minimal mediums, supplement 0.1mg/L 6-BA, 0.1mg/L NAA, 15g/L
Sucrose, 5g/L agar.
Embodiment 9
The tissue-cultured seedling micro-grafting method of Japanese maple orange dream described in the present embodiment, the difference with embodiment 1 are：
In step (3) tissue-cultured seedling micrografting, MS minimal mediums, supplement MS minimal mediums, supplement 0.6mg/L 6-BA,
0.06mg/L NAA, 20g/L sucrose, 5g/L agar.
Embodiment 10
The tissue-cultured seedling micro-grafting method of Japanese maple orange dream described in the present embodiment, the difference with embodiment 1 are：
In step (3) tissue-cultured seedling micrografting, MS minimal mediums, supplement MS minimal mediums, supplement 0.6mg/L 6-BA,
0.05mg/L NAA, 15g/L sucrose, 5g/L agar.
Embodiment 11
The tissue-cultured seedling micro-grafting method of Japanese maple orange dream described in the present embodiment, the difference with embodiment 1 are：
In step (4) acclimatization and transplantses, micrografting seedling obtained by step (3) is transferred to greenhouse hardening at initial stage 6 days, then be transferred to seedbed
In the middle week of hardening 4-5, seedbed was carried out disinfection in 7 days in advance, the matrix in seedbed is that volume ratio is 2：2 perlite and humus
Mixture, in the seedbed during hardening, carrying out spraying treatment and sterilization processing, hardening can transplant after terminating, wherein institute
The method for stating spraying treatment is：During fine day, at interval of 10min sprayings 4s；Rainy day or it is cloudy when, at interval of 18min spray 3s, spray
The time of mist processing is 9 days, and the bactericidal treatments are：Grafting was sprayed comprehensively using microbicide solution every 6 days, it is described
Microbicide solution is 70% thiophanate methyl, 800 times of solution.
Embodiment 12
The tissue-cultured seedling micro-grafting method of Japanese maple orange dream described in the present embodiment, the difference with embodiment 1 are：
In step (4) acclimatization and transplantses, micrografting seedling obtained by step (3) is transferred to greenhouse hardening at initial stage 8 days, then be transferred to seedbed
In the middle week of hardening 4-5, seedbed was carried out disinfection in 7 days in advance, the matrix in seedbed is that volume ratio is 1：2 perlite and humus
Mixture, in the seedbed during hardening, carrying out spraying treatment and sterilization processing, hardening can transplant after terminating, wherein institute
The method for stating spraying treatment is：During fine day, at interval of 15min sprayings 3s；Rainy day or it is cloudy when, at interval of 25min spray 4s, spray
The time of mist processing is 8 days, and the bactericidal treatments are：Grafting was sprayed comprehensively using microbicide solution every 8 days, it is described
Microbicide solution is 70% thiophanate methyl, 800 times of solution.
Embodiment 13
The tissue-cultured seedling micro-grafting method of Japanese maple orange dream described in the present embodiment, the difference with embodiment 1 are：
In step (4) acclimatization and transplantses, micrografting seedling obtained by step (3) is transferred to greenhouse hardening at initial stage 5 days, then be transferred to seedbed
In the middle week of hardening 4-5, seedbed was carried out disinfection in 7 days in advance, the matrix in seedbed is that volume ratio is 2：3 perlite and humus
Mixture, in the seedbed during hardening, carrying out spraying treatment and sterilization processing, hardening can transplant after terminating, wherein institute
The method for stating spraying treatment is：During fine day, at interval of 15min sprayings 3s；Rainy day or it is cloudy when, at interval of 18min spray 4s, spray
The time of mist processing is 12 days, and the bactericidal treatments are：Grafting was sprayed comprehensively using microbicide solution every 5 days, institute
It is 70% thiophanate methyl, 800 times of solution to state microbicide solution.
Embodiment 14
The tissue-cultured seedling micro-grafting method of Japanese maple orange dream described in the present embodiment, the difference with embodiment 1 are：
In step (4) acclimatization and transplantses, micrografting seedling obtained by step (3) is transferred to greenhouse hardening at initial stage 7 days, then be transferred to seedbed
In the middle week of hardening 4-5, seedbed was carried out disinfection in 7 days in advance, the matrix in seedbed is that volume ratio is 2：3 perlite and humus
Mixture, in the seedbed during hardening, carrying out spraying treatment and sterilization processing, hardening can transplant after terminating, wherein institute
The method for stating spraying treatment is：During fine day, at interval of 10min sprayings 3s；Rainy day or it is cloudy when, at interval of 20min spray 3s, spray
The time of mist processing is 10 days, and the bactericidal treatments are：Grafting was sprayed comprehensively using microbicide solution every 7 days, institute
It is 50% carbendazim, 800 times of solution to state microbicide solution.
Embodiment 15
The tissue-cultured seedling micro-grafting method of Japanese maple orange dream described in the present embodiment, the difference with embodiment 1 are：
In step (4) acclimatization and transplantses, micrografting seedling obtained by step (3) is transferred to greenhouse hardening at initial stage 7 days, then be transferred to seedbed
In the middle week of hardening 4-5, seedbed was carried out disinfection in 7 days in advance, the matrix in seedbed is that volume ratio is 2：3 perlite and humus
Mixture, in the seedbed during hardening, carrying out spraying treatment and sterilization processing, hardening can transplant after terminating, wherein institute
The method for stating spraying treatment is：During fine day, at interval of 10min sprayings 3s；Rainy day or it is cloudy when, at interval of 20min spray 3s, spray
The time of mist processing is 6 days, and the bactericidal treatments are：Grafting was sprayed comprehensively using microbicide solution every 7 days, it is described
Microbicide solution is 75% Bravo, 800 times of solution.
Embodiment 16
The grafting wound callus growth time of the grafting of the tissue culture micro-grafting method of difference Statistics Implementation example 1-15,
Grafting wound healing live time, survival rate, graftable time, growing way, the results are shown in Table 1.
The tissue culture micro-grafting method result of 1 embodiment of table 1-15 counts
Note："+" represents grafting growing way in table, and "+", and to represent growing way better.
As can be seen from the above table, the method for the tissue-cultured seedling micrografting of embodiment 1-15, its grafting wound healing is fast, and into
Motility rate is higher, and growing way is also preferable, improves the quality of grafting, and the tissue-cultured seedling micro-grafting method of the application can enter in whole year
OK, not by environment and season limit, the graftable time greatly prolongs, thus also improves the yield of grafting, during tissue-cultured seedling culture
Between it is short, micrografting seedling incubation time is short, substantially increases the efficiency of breeding, overcomes traditional engrafting method and is limited by season and environment
The defects of system, grafting time is shorter, and operating technology requires high, and grafting efficiency is low, and reproductive number is few.Can be with from embodiment 1-10
Find out, in tissue-cultured seedling micrografting step, when the composition of culture medium is the combination condition of auxin, sucrose, agar, it is hindered
Mouth healing faster, and supplements 1.0mg/L 6-BA, 0.05mg/L NAA, 20g/L sucrose in MS minimal mediums, 5g/L agar
When, its grafting wound healing time and wound callus growth time are most short, are optimal medium condition.Comparative example
1-15, it can be seen that the grafting wound healing time and wound callus for the grafting that the technical scheme of embodiment 1 obtains
Growth time faster, survival rate highest, and growing way is preferable, is the preferable scheme of the application.Compared to traditional engrafting method, this Shen
Please technical scheme micro- potted landscape, micro- potted plant can be used for the smaller grafting of production specification, solve tissue culture and take root difficult problem, it is bred
Speed is faster than tradition, small by seasonal effect.
It the above is only the preferred embodiment of the present invention, it is noted that above-mentioned preferred embodiment is not construed as pair
The limitation of the present invention, protection scope of the present invention should be defined by claim limited range.For the art
For those of ordinary skill, without departing from the spirit and scope of the present invention, some improvements and modifications can also be made, these change
Enter and retouch and also should be regarded as protection scope of the present invention.
Claims (1)
- A kind of 1. tissue-cultured seedling micro-grafting method of Japanese maple orange dream, it is characterised in that：It the described method comprises the following steps：(1) sterile stock is cultivated：Cut Fructus Aceris elegantulis band bud branch section, carry out tissue cultures, obtain Fructus Aceris elegantulis aseptic seedling, by Fructus Aceris elegantulis without Vaccine beveling is blocked, and obtains sterile stock, the stem of the sterile stock is slightly more than 1.5mm；(2) sterile scion cultivated：Japanese maple orange dream band bud branch section is cut, tissue cultures is carried out, it is sterile to obtain Japanese maple orange dream Seedling, the beveling of Japanese maple orange dream aseptic seedling is blocked, obtains sterile scion, the stem of the sterile scion is slightly more than 1mm；(3) tissue-cultured seedling micrografting：By the oblique of the oblique section of the sterile stock described in step (1) and step (2) the sterile scion Section is alignd, and fixed, obtains micrografting seedling, and the grafting is inoculated in into the week of culture 4-5 on culture medium, and temperature is 23- 27 DEG C, intensity of illumination is 2300-2600Lx, light application time 16h/d, and the composition of the culture medium is：MS minimal mediums, 1.0mg/L 6-BA, 0.05mg/L NAA, 20g/L sucrose, 3-7g/L agar are supplemented, wherein, the pH value of the agar is 6.5；(4) acclimatization and transplantses：Micrografting seedling obtained by step (3) is transferred to greenhouse hardening at initial stage 5-8 days, then is transferred to hardening in seedbed In 4-5 weeks, in the seedbed during hardening, carrying out spraying treatment and sterilization processing, hardening can transplant after terminating, wherein, institute The matrix for stating seedbed is that volume ratio is 2：2-4 perlite and the mixture of humus；The method of the spraying treatment is：It is fine It when, at interval of 10-15min spray 3-4s；Rainy day or it is cloudy when, at interval of 18-25min spray 3-4s, spraying treatment Time be 6-12 days；The bactericidal treatments are：Grafting was sprayed comprehensively using microbicide solution every 5-8 days, its In, the microbicide solution is molten selected from 70% thiophanate methyl, 800 times of solution, 800 times of 50% carbendazim for any one Liquid, 75% Bravo, 800 times of solution.
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