CN109392714A - A kind of tissue culture and rapid propagation method of Kiwi berry gold plum - Google Patents
A kind of tissue culture and rapid propagation method of Kiwi berry gold plum Download PDFInfo
- Publication number
- CN109392714A CN109392714A CN201811400370.9A CN201811400370A CN109392714A CN 109392714 A CN109392714 A CN 109392714A CN 201811400370 A CN201811400370 A CN 201811400370A CN 109392714 A CN109392714 A CN 109392714A
- Authority
- CN
- China
- Prior art keywords
- culture
- seedling
- illumination
- days
- adventitious
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- 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
- A01H4/001—Culture apparatus for tissue culture
-
- 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
Abstract
The invention discloses a kind of tissue culture and rapid propagation method of Kiwi berry gold plum, step includes: 1. to obtain aseptic seedling；2. adventitious shoot regeneration；3. adventitious bud proliferation, strong seedling culture；4. culture of rootage；5. transplanting；It is characterized by comprising germination medium, regeneration culture medium, proliferation strong seedling culture base and root media and corresponding condition of culture.The present invention carries out tissue cultures to golden plum using four kinds of specific aim culture mediums, the demand of a large amount of golden plum Kiwi berry seedling of fast breeding can be realized in a short time, and it can guarantee seedling quality to the maximum extent by tissue cultures, improve seedling breeding speed, it realizes seedling industrialized production, meets in production the needs of to golden plum seedling；Direct tissue culture kind seedling, establishes kind cutting orchard, breaks the bottleneck for grafting the amount of scion in traditional propagation by grafiting, sufficiently meets the needs of production.
Description
Technical field
The present invention relates to a kind of Kiwifruit Cultivars tissue culture and rapid propagation methods, more particularly to golden plum Kiwifruit Cultivars tissue-culturing rapid propagation side
Method.
Background technique
Jin Mei is that Wuhan Botanical Garden, Chinese Acadmey of Sciences was hybridized using ' Jin Yan ' with Chinese red-pulp kiwi fruit staminiferous plant in 2002,
The excellent strain filtered out from F1 generation group, by filial generation identification and regional testing breeding in 10 years, in application agriculture in 2013
Industry New variety protection.The long flat ellipse of fruit, it is up-small and down-big, about 90 grams of Mean Fruit Weight, the green and brown color of pericarp, fruit face down.Fruit
Meat yellow or yellow green, the dense sweet tea of taste give off a strong fragrance, and matter is tender, succulence, and mouthfeel is good.The soft ripe rear soluble solid of fruit averagely contains
16%, dry matter 15%, 125 milligrams/100 milligrams of Victoria C, mineral element is abundant, nitrogenous 0.15%, potassium 0.18%, calcium 34 milligrams/thousand
Gram, fruit is compared with storage endurance, and shelf life 5-7 days.Tree vigo(u)r is more prosperous, germination rate 58%, fruit branch rate 80%.It grafts the 4th annual strain and produces 38
Kilogram, high yield, stable yields.Gold plum source of seedling is predominantly grafted at present, obtains golden plum seedling in method and technology by tissue cultures
On still belong to blank, the method that the present invention uses tissue cultures is quickly and effectively saved under the premise of guaranteeing germplasm quality, is bred
Golden plum seedling realizes that its industrial seedling rearing produces, meets in the market the needs of to golden plum seedling.Existing market is mainly using grafting
Golden plum seedling is bred with cottage method, cuttage survival rate is low, grafts the limitation of Quantity of the scion, is widely applied Jin Mei and faces kind
The insufficient problem in seedling source.
Summary of the invention
In view of the deficiencies in the prior art, the object of the invention is that the tissue culture for providing a kind of Kiwi berry gold plum is fast
Breeding method, i.e., directly obtain that quality is uniform, excellent in vitro golden plum tissue culture seedling by method for tissue culture, and by optimization from
Body rapid propagation system realizes the purpose of fast breeding gold plum Kiwi berry high quality seedling, for ever-increasing seedling demand in production.
The technical solution adopted by the present invention is as follows:
Using golden plum annotinous branch water planting rudiment as explant, tissue cultures are carried out after disinfection, aseptic seedling are obtained, with sterile miaoye
Piece is regenerating tissues source, carries out induction differentiation, culture of rootage.Adventitious bud Direct Regeneration rate is up to 92%, and value-added coefficient is reachable
4.3.It can be quickly obtained a large amount of fine quality seedlings in a short time, realize golden plum seedling rapid propagation in factory.
Specifically, this method includes the following steps:
1. the acquisition of aseptic seedling
1) annotinous branch full from clip robust growth axillary bud on golden plum elite stand at the end of November, is cut into 30cm long shoot for branch
Section, every section of 5-10 bud；
2) the branch section morphology upper end being sealed with sealing film, indoor culture in 2 ‰ aqueous sucrose solutions is impregnated in morphology lower end,
25 ± 2 DEG C of environment temperature of holding, guarantee indoor cleaning, the aqueous solution of replacement in 2-3 days, after two weeks, axillary bud eruption；
3) when axillary bud sprout it is long to 1-2cm when, take young tender shoots body on superclean bench, being placed in sterilized empty bottle, 75% wine
Smart surface sterilization 1min, pours out alcohol, and it is that 0.1% mercuric chloride deeply sterilizes 6-10min that mass volume ratio, which is added, mercuric chloride solution and outer
Implant comes into full contact with concussion, guarantees that disinfection thoroughly, pours out thimerosal, and explant more changes in new sterilizing bottle, sterile water concussion
It rinses 4-5 times, is dried in sterilizing filter paper；
4) remove the wound of axillary bud sprout base portion and the blade of damage, be inoculated on explant germination medium；
5) after 30-40 days, sprout lamina, robust growth obtains aseptic seedling；
2. adventitious shoot regeneration induces
1) in cutting aseptic seedling young leaflet tablet on superclean bench, it is cut into 0.5*0.5cm leaf dish, is placed on regeneration culture medium, pH
5.8 are adjusted to, the regeneration induction of adventitious bud is carried out；
2) inoculation material is placed under illumination condition and cultivates；
3) after 40-50 days, blade Direct Regeneration adventitious bud；
3. adventitious bud proliferation, strong seedling culture
Regeneration obtains adventitious bud after subculture is primary on best adventitious shoot regeneration culture medium, and adventitious bud is cut into single sprout,
It is placed on proliferation strong seedling culture base and carries out proliferation strong seedling culture, pH is adjusted to 5.8；
4. culture of rootage
By the relatively uniform high 2-3cm of growth conditions, the adventitious bud for having 3-5 piece leaf cuts, is inoculated on root media and carries out
Culture of rootage, 8-10 days i.e. existing visually visible adventitious roots, short and thick, healthy and strong, root long 1.0-3.0cm after 25-30 days, 3-5 item root, i.e.,
It can transplant.
5. transplanting
1) after culture of rootage 25 days, bottle seedling of taking root is directly taken out, the culture medium in foundation is cleaned with tap water；
2) seedling is planted to the small nutritive cube that Nutrition Soil is housed, and nutritive cube top covered plastic film guarantees locating for new transplanting seedling
Ambient humidity be 85%-90%, temperature control removes plastic film after 25-30 DEG C, 10-15 days, ambient humidity suitably reduces
To 60%-75%；
3) conventional water and fertilizer management after 40-45 days, environment temperature control is at 35 DEG C hereinafter, high temperature summer need to shade and Wet-curtain temperature reducing；
It is characterized by:
The germination medium are as follows:+0.8% agar of MS+TDZ 0.5-3.0 mg/L+1%-2% sucrose；
In the preparation method of the aseptic seedling, inoculation material condition of culture are as follows: intensity of illumination 2400-2600Lx, illumination 12h/d,
25 ± 2 DEG C of room temperature of culture；
The leaf regeneration culture medium are as follows:+0.8% fine jade of MS+TDZ 0.5-2.0 mg/L+IAA 0.25-1.0 mg/L+3% sucrose
Rouge；
In the blade adventitious shoot regeneration abductive approach, inoculation material condition of culture are as follows: intensity of illumination 2600-3000Lx, illumination
12h/d cultivates 25 ± 2 DEG C of room temperature；
The proliferation strong seedling culture base are as follows:+0.8% fine jade of MS+TDZ 0.1-2.0 mg/L+IAA 0.01-0.1mg/L+3% sucrose
Rouge；
The adventitious bud proliferation, strong seedling culture method, inoculation material condition of culture are as follows: intensity of illumination 3200-3500Lx, illumination
12h/d cultivates 25 ± 2 DEG C of room temperature；
The root media are as follows: 1/2MS+ banana puree 2-15 g/L+IAA 0.1-1.0 mg/L+0.8-1.5% agar；
In the culture of rootage method, inoculation material condition of culture are as follows: intensity of illumination 3500-3800Lx, illumination 12h/d, culture
25 ± 2 DEG C of room temperature；
The Nutrition Soil ingredient are as follows: yellow soil: turfy soil: perlite=2:1:1.
Innovative point of the invention: the method that initial stage of the invention uses indoor aqueous sucrose solution culture, to inspire axillary bud sprouting
As explant source, evade the pollution sources of direct outdoor sampling, greatly reduces the wind that bacterium, Mycophyta and virus carry
Danger, while the mercuric chloride processing time is reduced, reduce the injury to explant histocyte physiologically.To material in tissue culture procedures
The intensity of illumination of receiving makes gradient adjustment, and domestication is transplanted to it and plays induction adaptation well.Late stage of culture is taken root
Stage increases the usage amount of agar powder, is more advantageous to root of hair, cleaning when also being more conducive to transplant to plant foundation position culture medium,
To reduce the damage to root, transplanting survival rate is greatly improved.The stage of taking root use banana puree instead of traditional sucrose as
Energy source, seedling early growth is healthy and strong, takes root short and thick, is conducive to later period transplanting survival rate.Seedling is directly transplanted without domestication, is saved
Time and manpower related resource, from largely reducing production cost.The control of temperature and humidity in transplanting initial stage certain time
System is conducive to seedling environment after rapidly adapting to transplanting in short term, both shortens breeding time, also greatly improve kind of a transplantation of seedlings
Survival rate.
The present invention has following advantages and good effect:
It, can a large amount of golden plum of fast breeding in a short time 1. the present invention carries out tissue cultures to golden plum using four kinds of specific aim culture mediums
Kiwi berry seedling, and can guarantee seedling quality to greatest extent by tissue cultures, seedling breeding speed is improved, realizes seedling
The factorial production meets in raw factory the needs of to golden plum seedling.
2. explant uses water planting budding mode during providing, can control to the greatest extent in aseptic seedling establishment process
Pollution, largely improve seedling breeding speed, shorten the seedling breeding period, reduce tissue culture cost.
3. during seedling fostering, the enhancing of the change of condition of culture, especially intensity of illumination is conducive to seedling accumulation
Nutrient growth ability enhances seedling resistance, greatly improves later period tissue culture seedling transplanting survival rate.
4. planting transplantation of seedlings early period, to the strict control of the temperature and humidity of transplanting environment, it can guarantee kind of a transplantation of seedlings to the greatest extent
Survival rate, to reduce loss, reduce production cost.
5. obtaining golden plum seedling relative to previous cuttage, grafting, tissue-culturing rapid propagation nursery more can guarantee seedling quality, seedling
Growth is consistent, neat, and can meet production within the shorter time to the quantitative demand of seedling.
6. direct tissue culture kind seedling, establishes kind cutting orchard, break the bottle that the amount of scion is grafted in traditional propagation by grafiting
Neck sufficiently meets the needs of production.
Specific embodiment
It is described in detail with reference to embodiments.
One, embodiment 1
A kind of tissue culture and rapid propagation method of Kiwi berry gold plum is completed in the steps below:
1, the acquisition of aseptic seedling
The annotinous branch full from clip robust growth axillary bud on golden plum elite stand at the end of November, is cut into 30cm long shoot section, and every section
5-10 bud, morphology upper end seal with sealing film, and indoor culture in 2 ‰ aqueous sucrose solutions is impregnated in morphology lower end, keep environment
25 ± 2 DEG C of temperature, guarantee that indoor cleaning, the aqueous solution of replacement in 2-3 days, after two weeks, axillary bud eruption are long to 1-2cm to sprout
When, take young tender shoots body on superclean bench, being placed in sterilized empty bottle, 75% alcohol surface sterilization 1min pours out alcohol,
It is that 0.1% mercuric chloride deeply sterilizes 6min that mass volume ratio, which is added, and mercuric chloride solution fullys shake with external body, guarantees disinfection thoroughly,
Thimerosal out, explant more change in new sterilizing bottle, and sterile water concussion is rinsed 4-5 times, dry, remove in sterilizing filter paper
The wound of sprout base portion and the blade of damage are inoculated in 2.0 mg/L+2% sucrose+0.8% of explant germination medium MS+TDZ
On agar, after 30 days, sprout lamina, robust growth obtains aseptic seedling.Inoculation material is cultivated under illumination condition, intensity of illumination
2400Lx cultivates 25 ± 2 DEG C of room temperature, illumination 12h/d.
2, adventitious shoot regeneration induces
In cutting tests for sterility on superclean bench, it is cut into 0.5*0.5cm leaf dish, is placed in regeneration culture medium MS+TDZ 1.0
On+0.8% agar of 0.25 mg/L+3% sucrose of mg/L+IAA, pH is adjusted to 5.8, carries out the regeneration induction of adventitious bud.Inoculation material
It is placed under illumination condition and cultivates.Intensity of illumination 2600Lx cultivates 25 ± 2 DEG C of room temperature, illumination 12h/d.After 45 days, blade is straight
Regenerated adventitious bud is connect, quantity is more, and is clump bud, and differentiation is fast, and color bright green, adventitious bud Direct Regeneration is up to 92%.
3, adventitious bud proliferation, strong seedling culture
Regeneration obtains adventitious bud after subculture is primary on best adventitious shoot regeneration culture medium, and adventitious bud is cut into single sprout,
It is placed on+0.8% agar medium of 0.5 mg/L+IAA of MS+TDZ, 0.02 mg/L+3% sucrose and carries out proliferation strong seedling culture, pH
It is adjusted to 5.8.Intensity of illumination 3200Lx cultivates 25 ± 2 DEG C of room temperature, illumination 12h/d.
4, culture of rootage
By the relatively uniform high 2-3cm of growth conditions, the adventitious bud for having 3-5 piece leaf cuts, is inoculated on root media and carries out
Sucrose is not added in culture medium in culture of rootage, 0.8 mg/L+1.2% agar of 1/2MS+ banana puree 12g/L+IAA, does not have to adjust pH,
Intensity of illumination 3800Lx cultivates 25 ± 2 DEG C of room temperature, and illumination 12h/d, 8 days i.e. existing visually visible adventitious roots are short and thick, healthy and strong,
Root long 1.0-3.0cm after 28 days, 3-5 item root, can transplant.
5, it transplants
After culture of rootage 25 days, bottle seedling of taking root is directly taken out, cleans the culture medium in foundation with tap water, seedling is planted to equipped with battalion
Support the small nutritive cube of soil, Nutrition Soil ingredient are as follows: yellow soil: turfy soil: perlite=2:1:1.Add plastic film above nutritive cube,
Guarantee that ambient humidity locating for new transplanting seedling is 90%, plastic film, environmental wet are removed in temperature control after 25-30 DEG C, 15 days
Degree is suitably reduced to 75%.Conventional water and fertilizer management after 45 days, environment temperature control is at 35 DEG C hereinafter, high temperature summer need to shade and wet
Curtain cooling.
High survival rate is counted after two months up to 97%.
Two, embodiment 2
A kind of tissue culture and rapid propagation method of Kiwi berry gold plum is completed in the steps below:
1, the acquisition of aseptic seedling
The annotinous branch full from clip robust growth axillary bud on golden plum elite stand at the end of November, is cut into 30cm long shoot section, and every section
5-10 bud, morphology upper end seal with sealing film, and indoor culture in 2 ‰ aqueous sucrose solutions is impregnated in morphology lower end, keep environment
25 ± 2 DEG C of temperature, guarantee that indoor cleaning, the aqueous solution of replacement in 2-3 days, after two weeks, axillary bud eruption are long to 1-2cm to sprout
When, take young tender shoots body on superclean bench, being placed in sterilized empty bottle, 75% alcohol surface sterilization 1min pours out alcohol,
It is that 0.1% mercuric chloride deeply sterilizes 8min that mass volume ratio, which is added, and mercuric chloride solution fullys shake with external body, guarantees disinfection thoroughly,
Thimerosal out, explant more change in new sterilizing bottle, and sterile water concussion is rinsed 4-5 times, dry, remove in sterilizing filter paper
The wound of sprout base portion and the blade of damage are inoculated in 0.5 mg/L+2% sucrose+0.8% of explant germination medium MS+TDZ
On agar, after 30 days, sprout lamina, robust growth obtains aseptic seedling.Inoculation material is cultivated under illumination condition, intensity of illumination
2500Lx cultivates 25 ± 2 DEG C of room temperature, illumination 12h/d.
2, adventitious shoot regeneration induces
In cutting tests for sterility on superclean bench, it is cut into 0.5*0.5cm leaf dish, is placed in regeneration culture medium MS+TDZ 0.5
On+0.8% agar of 0.5 mg/L+3% sucrose of mg/L+IAA, pH is adjusted to 5.8, carries out the regeneration induction of adventitious bud.Inoculation material is set
It is cultivated under illumination condition.Intensity of illumination 2800Lx cultivates 25 ± 2 DEG C of room temperature, illumination 12h/d.After 40 days, blade is direct
Regenerated adventitious bud, quantity is more, and is clump bud, and differentiation is fast, and color bright green, adventitious bud Direct Regeneration is up to 90%.
3, adventitious bud proliferation, strong seedling culture
Regeneration obtains adventitious bud after subculture is primary on best adventitious shoot regeneration culture medium, and adventitious bud is cut into single sprout,
It is placed on+0.8% agar medium of 0.1 mg/L+IAA 0.01mg/L+3% sucrose of MS+TDZ and carries out proliferation strong seedling culture, pH tune
To 5.8.Intensity of illumination 3400Lx cultivates 25 ± 2 DEG C of room temperature, illumination 12h/d.
4, culture of rootage
By the relatively uniform high 2-3cm of growth conditions, the adventitious bud for having 3-5 piece leaf cuts, is inoculated on root media and carries out
Culture of rootage, 0.1 mg/L+0.8% agar of 1/2MS+ banana puree 2g/L+IAA do not have to addition sucrose, pH5.8.Intensity of illumination
3600Lx cultivates 25 ± 2 DEG C of room temperature, and illumination 12h/d, 8 days are the visible adventitious root of existing naked eyes, short and thick, healthy and strong, root after 25 days
Long 1.0-3.0cm, 3-5 item root, can transplant.
5, it transplants
After culture of rootage 25 days, bottle seedling of taking root is directly taken out, cleans the culture medium in foundation with tap water, seedling is planted to equipped with battalion
Support the small nutritive cube of soil, Nutrition Soil ingredient are as follows: yellow soil: turfy soil: perlite=2:1:1.Add plastic film above nutritive cube,
Guarantee that ambient humidity locating for new transplanting seedling is 88%, plastic film, environmental wet are removed in temperature control after 25-30 DEG C, 10 days
Degree is suitably reduced to 75%.Conventional water and fertilizer management after 45 days, environment temperature control is at 35 DEG C hereinafter, high temperature summer need to shade and wet
Curtain cooling.
High survival rate is counted after two months up to 95%.
Three, embodiment 3
A kind of tissue culture and rapid propagation method of Kiwi berry gold plum is completed in the steps below:
1, the acquisition of aseptic seedling
The annotinous branch full from clip robust growth axillary bud on golden plum elite stand at the end of November, is cut into 30cm long shoot section, and every section
5-10 bud, morphology upper end seal with sealing film, and indoor culture in 2 ‰ aqueous sucrose solutions is impregnated in morphology lower end, keep environment
25 ± 2 DEG C of temperature, guarantee that indoor cleaning, the aqueous solution of replacement in 2-3 days, after two weeks, axillary bud eruption are long to 1-2cm to sprout
When, take young tender shoots body on superclean bench, being placed in sterilized empty bottle, 75% alcohol surface sterilization 1min pours out alcohol,
It is that 0.1% mercuric chloride deeply sterilizes 10min that mass volume ratio, which is added, and mercuric chloride solution fullys shake with external body, guarantees disinfection thoroughly,
Thimerosal is poured out, explant more changes in new sterilizing bottle, and sterile water concussion is rinsed 4-5 times, dries, goes in sterilizing filter paper
Fall the wound of sprout base portion and the blade of damage, is inoculated in explant germination medium MS+TDZ3.0 mg/L+2% sucrose+0.8%
On agar, after 38 days, sprout lamina, robust growth obtains aseptic seedling.Inoculation material is cultivated under illumination condition, intensity of illumination
2600Lx cultivates 25 ± 2 DEG C of room temperature, illumination 12h/d.
2, adventitious shoot regeneration induces
In cutting tests for sterility on superclean bench, it is cut into 0.5*0.5cm leaf dish, is placed in regeneration culture medium MS+TDZ 2.0
On+0.8% agar of mg/L+IAA1.0 mg/L+3% sucrose, pH is adjusted to 5.8, carries out the regeneration induction of adventitious bud.Inoculation material is set
It is cultivated under illumination condition.Intensity of illumination 3000Lx cultivates 25 ± 2 DEG C of room temperature, illumination 12h/d.After 48 days, blade is direct
Regenerated adventitious bud, quantity is more, and is clump bud, and differentiation is fast, and color bright green, adventitious bud Direct Regeneration is up to 91.7%.
3, adventitious bud proliferation, strong seedling culture
Regeneration obtains adventitious bud after subculture is primary on best adventitious shoot regeneration culture medium, and adventitious bud is cut into single sprout,
It is placed on+0.8% agar medium of 0.1 mg/L+3% sucrose of MS+TDZ2.0 mg/L+IAA and carries out proliferation strong seedling culture, pH tune
To 5.8.Intensity of illumination 3400Lx cultivates 25 ± 2 DEG C of room temperature, illumination 12h/d.
4, culture of rootage
By the relatively uniform high 2-3cm of growth conditions, the adventitious bud for having 3-5 piece leaf cuts, is inoculated on root media and carries out
Culture of rootage, 15 g/L+IAA of 1/2MS+ banana puree, 1.0 mg/L+1.5% agar do not have to addition sucrose, do not adjust pH.Illumination
Intensity 3500Lx cultivates 25 ± 2 DEG C of room temperature, and illumination 12h/d, 10 days i.e. existing visually visible adventitious roots are short and thick, healthy and strong, and 25
Root long 1.0-3.0cm after it, 3-5 item root, can transplant.
5, it transplants
After culture of rootage 25 days, bottle seedling of taking root is directly taken out, cleans the culture medium in foundation with tap water, seedling is planted to equipped with battalion
Support the small nutritive cube of soil, Nutrition Soil ingredient are as follows: yellow soil: turfy soil: perlite=2:1:1.Add plastic film above nutritive cube,
Guarantee that ambient humidity locating for new transplanting seedling is 85%, plastic film, environmental wet are removed in temperature control after 25-30 DEG C, 15 days
Degree is suitably reduced to 65%.Conventional water and fertilizer management after 45 days, environment temperature control is at 35 DEG C hereinafter, high temperature summer need to shade and wet
Curtain cooling.High survival rate is counted after two months up to 94.8%.
Claims (1)
1. a kind of tissue culture and rapid propagation method of Kiwi berry gold plum, which comprises the following steps:
1. the acquisition of aseptic seedling
1) annotinous branch full from clip robust growth axillary bud on golden plum elite stand at the end of November, is cut into 30cm long shoot for branch
Section, every section of 5-10 bud；
2) the branch section morphology upper end being sealed with sealing film, indoor culture in 2 ‰ aqueous sucrose solutions is impregnated in morphology lower end,
25 ± 2 DEG C of environment temperature of holding, guarantee indoor cleaning, the aqueous solution of replacement in 2-3 days, after two weeks, axillary bud eruption；
3) when axillary bud sprout it is long to 1-2cm when, take young tender shoots body on superclean bench, being placed in sterilized empty bottle, 75% wine
Smart surface sterilization 1min, pours out alcohol, and it is that 0.1% mercuric chloride deeply sterilizes 6-10min that mass volume ratio, which is added, mercuric chloride solution and outer
Implant comes into full contact with concussion, guarantees that disinfection thoroughly, pours out thimerosal, and explant more changes in new sterilizing bottle, sterile water concussion
It rinses 4-5 times, is dried in sterilizing filter paper；
4) remove the wound of axillary bud sprout base portion and the blade of damage, be inoculated on explant germination medium；
5) after 30-40 days, sprout lamina, robust growth obtains aseptic seedling；
2. adventitious shoot regeneration induces
1) in cutting aseptic seedling young leaflet tablet on superclean bench, it is cut into 0.5*0.5cm leaf dish, is placed on regeneration culture medium, pH
5.8 are adjusted to, the regeneration induction of adventitious bud is carried out；
2) inoculation material is placed under illumination condition and cultivates；
3) after 40-50 days, blade Direct Regeneration adventitious bud；
3. adventitious bud proliferation, strong seedling culture
Regeneration obtains adventitious bud after subculture is primary on best adventitious shoot regeneration culture medium, and adventitious bud is cut into single sprout,
It is placed on proliferation strong seedling culture base and carries out proliferation strong seedling culture, pH is adjusted to 5.8；
4. culture of rootage
By the relatively uniform high 2-3cm of growth conditions, the adventitious bud for having 3-5 piece leaf cuts, is inoculated on root media and carries out
Culture of rootage, 8-10 days i.e. existing visually visible adventitious roots, short and thick, healthy and strong, root long 1.0-3.0cm after 25-30 days, 3-5 item root, i.e.,
It can transplant；
5. transplanting
1) after culture of rootage 25 days, bottle seedling of taking root is directly taken out, the culture medium in foundation is cleaned with tap water；
2) seedling is planted to the small nutritive cube that Nutrition Soil is housed, and nutritive cube top covered plastic film guarantees locating for new transplanting seedling
Ambient humidity be 85%-90%, temperature control removes plastic film after 25-30 DEG C, 10-15 days, ambient humidity suitably reduces
To 60%-75%；
3) conventional water and fertilizer management after 40-45 days, environment temperature control is at 35 DEG C hereinafter, high temperature summer need to shade and Wet-curtain temperature reducing；
It is characterized by:
The germination medium are as follows:+0.8% agar of MS+TDZ 0.5-3.0 mg/L+1%-2% sucrose；
In the preparation method of the aseptic seedling, inoculation material condition of culture are as follows: intensity of illumination 2400-2600Lx, illumination 12h/d,
25 ± 2 DEG C of room temperature of culture；
The leaf regeneration culture medium are as follows:+0.8% fine jade of MS+TDZ 0.5-2.0 mg/L+IAA 0.25-1.0 mg/L+3% sucrose
Rouge；
In the blade adventitious shoot regeneration abductive approach, inoculation material condition of culture are as follows: intensity of illumination 2600-3000Lx, illumination
12h/d cultivates 25 ± 2 DEG C of room temperature；
The proliferation strong seedling culture base are as follows:+0.8% fine jade of MS+TDZ 0.1-2.0 mg/L+IAA 0.01-0.1mg/L+3% sucrose
Rouge；
The adventitious bud proliferation, strong seedling culture method, inoculation material condition of culture are as follows: intensity of illumination 3200-3500Lx, illumination
12h/d cultivates 25 ± 2 DEG C of room temperature；
The root media are as follows: 1/2MS+ banana puree 2-15 g/L+IAA 0.1-1.0 mg/L+0.8-1.5% agar；
In the culture of rootage method, inoculation material condition of culture are as follows: intensity of illumination 3500-3800Lx, illumination 12h/d, culture
25 ± 2 DEG C of room temperature；
The Nutrition Soil ingredient are as follows: yellow soil: turfy soil: perlite=2:1:1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811400370.9A CN109392714A (en) | 2018-11-22 | 2018-11-22 | A kind of tissue culture and rapid propagation method of Kiwi berry gold plum |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811400370.9A CN109392714A (en) | 2018-11-22 | 2018-11-22 | A kind of tissue culture and rapid propagation method of Kiwi berry gold plum |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109392714A true CN109392714A (en) | 2019-03-01 |
Family
ID=65474589
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811400370.9A Pending CN109392714A (en) | 2018-11-22 | 2018-11-22 | A kind of tissue culture and rapid propagation method of Kiwi berry gold plum |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109392714A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111280004A (en) * | 2019-11-26 | 2020-06-16 | 广西壮族自治区农业科学院 | Asparagus cochinchinensis planting method |
| CN111758406A (en) * | 2020-06-28 | 2020-10-13 | 中国科学院武汉植物园 | Grafting preservation method for kiwi fruit in-vitro resources |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102177847A (en) * | 2011-03-07 | 2011-09-14 | 沈阳农业大学 | Factory seedling cultivating method of actinidia arguta |
| CN105557523A (en) * | 2015-12-16 | 2016-05-11 | 濮阳职业技术学院 | Establishment method of kiwi fruit genetic transformation receptor system based on heat stress |
| CN108077068A (en) * | 2016-11-21 | 2018-05-29 | 蒲江橙海阳光农业科技有限公司 | A kind of rapid propagation method of the sterile seedling of Kiwi berry |
| CN108739384A (en) * | 2018-05-31 | 2018-11-06 | 福建省农业科学院果树研究所 | A kind of method of Kiwifruit tissue-cultured seedling strengthening seedling and rooting |
-
2018
- 2018-11-22 CN CN201811400370.9A patent/CN109392714A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102177847A (en) * | 2011-03-07 | 2011-09-14 | 沈阳农业大学 | Factory seedling cultivating method of actinidia arguta |
| CN105557523A (en) * | 2015-12-16 | 2016-05-11 | 濮阳职业技术学院 | Establishment method of kiwi fruit genetic transformation receptor system based on heat stress |
| CN108077068A (en) * | 2016-11-21 | 2018-05-29 | 蒲江橙海阳光农业科技有限公司 | A kind of rapid propagation method of the sterile seedling of Kiwi berry |
| CN108739384A (en) * | 2018-05-31 | 2018-11-06 | 福建省农业科学院果树研究所 | A kind of method of Kiwifruit tissue-cultured seedling strengthening seedling and rooting |
Non-Patent Citations (4)
| Title |
|---|
| 于丽杰等: "《植物组织培养教程》", 31 July 2015, 华中科技大学出版社 * |
| 吕海燕等: ""‘东红’猕猴桃高效再生体系的建立"", 《广西植物》 * |
| 尚霄丽: ""猕猴桃、枣高效再生体系的建立及农杆菌介导的遗传转化研究"", 《中国博士学位论文全文数据库 农业科技辑》 * |
| 陈永勤等: ""一种美味猕猴桃‘金魁’快速无性繁殖方法"", 《湖北大学学报(自然科学版)》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111280004A (en) * | 2019-11-26 | 2020-06-16 | 广西壮族自治区农业科学院 | Asparagus cochinchinensis planting method |
| CN111758406A (en) * | 2020-06-28 | 2020-10-13 | 中国科学院武汉植物园 | Grafting preservation method for kiwi fruit in-vitro resources |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104782486B (en) | Tissue culture and intermediate propagation method for succulent Haworthia cooperivar. pilfera M. B. Bayer | |
| CN109392715A (en) | A kind of tissue culture and rapid propagation method of Kiwi berry gold and jade | |
| CN109392712A (en) | A kind of tissue culture and rapid propagation method of tara vine kind | |
| CN105706900A (en) | Sterile sowing and seedling raising method for hybrid orchid and Cymbidium tracyanum hybrid seeds | |
| CN107455261B (en) | A kind of method of Acer saccharum tissue culture quick breeding | |
| CN111616052A (en) | Rapid propagation and sugar-free rooting culture method and application of apple rootstock catalpa bungei | |
| CN104719163A (en) | Method for increasing acclimatization planting percent of blueberry tissue culture test-tube seedlings | |
| JP2008220242A (en) | Method for growing seedling of plant belonging to genus vaccininum | |
| CN108419675A (en) | A kind of tissue culture method of violet passion fruit top tip | |
| CN109392714A (en) | A kind of tissue culture and rapid propagation method of Kiwi berry gold plum | |
| CN107197746A (en) | A kind of mating system of China fir field excellent resources | |
| CN109042330A (en) | A kind of method for tissue culture of spindle tree | |
| CN108782252A (en) | A kind of method of micrografting in fresh water Chinese pear tissue-cultured seedling test tube | |
| CN109392713A (en) | A kind of tissue culture and rapid propagation method that Kiwi berry east is red | |
| CN110199886A (en) | A method of utilizing tissue culture technology micro cuttage tatarian dogwood | |
| CN107006367B (en) | 'sunshine' cherry tissue culture rapid propagation method | |
| CN110100740A (en) | A kind of tissue culture and rapid propagation method of two discriminations Platycerium bifurcatum | |
| CN109156361A (en) | A kind of in vitro quick breeding by group culture method of pruning Jia Opulus | |
| CN106550875B (en) | For the MS culture mediums of rouge radish tissue cultures, the quick breeding method for tissue culture of adventitious shoots culture base and rouge radish | |
| CN104686358A (en) | Sorbus alnifolia tissue culture and rapid propagation method | |
| CN101611698B (en) | Method for culturing cephalotaxus excised embryos and regenerating plants | |
| CN108184462A (en) | A kind of open country eggplant sapling stock breeding method and its method for grafting fruits and vegetables | |
| CN108450335A (en) | A kind of fresh water sand pear stem section tissue rapid propagation method | |
| CN108094207A (en) | Wet-land pine tree somatic embryo occurs and the method for plant regeneration | |
| CN111194693B (en) | Tissue culture rapid propagation method for new variety of Chinese flowering crabapple |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20190301 |
|
| RJ01 | Rejection of invention patent application after publication |