CN109105233A - The copper coin tree seedling fostering method and culture medium quickly bred based on micro cuttage - Google Patents
The copper coin tree seedling fostering method and culture medium quickly bred based on micro cuttage Download PDFInfo
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- CN109105233A CN109105233A CN201810898953.2A CN201810898953A CN109105233A CN 109105233 A CN109105233 A CN 109105233A CN 201810898953 A CN201810898953 A CN 201810898953A CN 109105233 A CN109105233 A CN 109105233A
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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
- A01G31/00—Soilless cultivation, e.g. hydroponics
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
- A01C1/08—Immunising seed
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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
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/10—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material
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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
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/30—Growth substrates; Culture media; Apparatus or methods therefor based on or containing synthetic organic compounds
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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
- A01H4/001—Culture apparatus for tissue culture
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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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Abstract
A kind of copper coin tree seedling fostering method quickly bred based on micro cuttage and culture medium, it include to grow the micro cuttage culture medium of lateral bud and root media for growing root system for the stem section of seedling parent for the stem section of seedling parent, pass through micro cuttage culture medium and root media, the parent of seedling is set to carry out segmentation fast breeding, direct nursery is no longer carried out by copper coin tree species, therefore realizes copper coin tree test tube seedling that is a large amount of, quickly, persistently obtaining high quality.
Description
One, technical field
The present invention relates to a kind of copper coin tree seedling fostering method and culture mediums, especially a kind of quickly to be bred based on micro cuttage
Copper coin tree seedling fostering method and culture medium.
Two, background technique
Copper coin tree (PaliurnsHemsleyanusRehd.) also known as bird not place, Rhamnaceae Paliurus deciduous tree, copper
Having many advantages, such as that survival rate is higher, growth is fast, result is early, anti-jujube witches broom after money tree grafting jujube tree, copper coin tree bark contains tannin,
Tannin extract, economic value, the ecological value and scientific research value with higher, therefore the copper coin quickly bred based on micro cuttage can be refined
Tree seedling fostering method and culture medium are a kind of important gardening methods, in the existing copper coin tree quickly bred based on micro cuttage
It is also all that nursery is carried out by copper coin tree species in seedling fostering method and culture medium, since outer kind of the shell of copper coin tree species is hard,
Cause seed water suction difficult, germination percentage is low, and big using the variation of the seedling group of seminal propagation, and uniformity is poor, and there are no one
The copper coin tree seedling fostering method quickly bred based on micro cuttage and culture medium are planted, is able to solve in copper coin tree seminal propagation seedling
The technical problem that germination percentage is low, reproduction speed is slow realizes the extensive fast breeding of copper coin tree species seedling, realize it is a large amount of, quickly, hold
The continuous copper coin tree test tube seedling for obtaining high quality.
Based on applicant's technical problem existing in the Disclosure of invention on May 15th, 2018 and background technique, technology
Feature and technical effect make application technical solution of the invention.
Three, summary of the invention
Object of the invention is a kind of copper coin tree seedling fostering method quickly bred based on micro cuttage,
Object of the invention is a kind of copper coin tree species seedling culture medium quickly bred based on micro cuttage.
In order to overcome the technical drawbacks described above, the object of the present invention is to provide a kind of copper coin trees quickly bred based on micro cuttage
Seedling fostering method and culture medium, therefore realize copper coin tree test tube seedling that is a large amount of, quickly, persistently obtaining high quality.
In order to achieve the above objectives, the technical solution adopted by the present invention is that: a kind of copper coin tree quickly bred based on micro cuttage
Seedling culture base includes to grow the micro cuttage culture medium of lateral bud and for for seedling parent for the stem section of seedling parent
Stem section grows the root media of root system.
Due to devising micro cuttage culture medium and root media, by micro cuttage culture medium and root media, make to plant
The parent of seedling carries out segmentation fast breeding, no longer carries out direct nursery by copper coin tree species, thus realize it is a large amount of, quickly, hold
The continuous copper coin tree test tube seedling for obtaining high quality.
The present invention devises, and also includes the germination culture medium for cultivating seedling parent.
The present invention devises, germination culture medium be set as MS minimal medium, micro cuttage culture medium be set as MS+1~
2mg/L 6-BA+0.1~0.4mg/L IBA+30g/L sucrose, root media are set as 1/4MS+0.5~1.5mg/L IBA+
30g/L sucrose.
The present invention devises, MS (Murashige and Skoog) culture medium composition:
1/4MS minimal medium composition:
2 plant growth regulating substances: 6- benzylaminopurine (6-BA), indolebutyric acid (IBA).
3 carbon sources: sucrose
4 deionized waters
5 agar powders.
The present invention devises, and a kind of copper coin tree seedling fostering method quickly bred based on micro cuttage the steps include: one
Stem section to the copper coin tree seedling of two sections carries out promoting bud growth in micro cuttage culture medium, middle stem section is obtained, middle stem section
Promoting root growth growth is carried out in root media, obtains copper coin tree species seedling.
The present invention devises, and the steps include:
Condition of culture: all culture mediums in cultural aim adjust pH value to 5.8 before sterilization, 1 big then at 121 DEG C
High pressure sterilization 20min under air pressure.Culturing room's cultivation temperature is 26 ± 2 DEG C, and periodicity of illumination is 16 hours/day,
One, the acquisition of aseptic seedling: being put into copper coin tree species in 10%KOH solution, the room temperature item for being 25 ± 2 DEG C in temperature
Sofening treatment is carried out to the kind skin of copper coin tree species under part, the soaking time in 10%KOH solution is set as 18-22min, soft
The copper coin tree species son distilled water flushing for changing processing is clean, then copper coin tree species of sofening treatment is put into 5% hypochlorous acid
Sodium solution carries out immersion treatment 8-12min, and sterile water is issued to after aseptic water washing three to five times and carries out seed soaking, when seed soaking
Between be set as 24-36h, and selecting copper coin tree species that expands of water suction is cultivation copper coin tree species, cultivation with copper coin tree species
Son is put into the first time disinfection treatment for carrying out that the time is 28-32s in 70% alcohol, the cultivation copper coin of first time disinfection treatment
Tree species is put into second of disinfection treatment for carrying out that the time is 8-12min in 5% liquor natrii hypochloritis, with sterile water to second
Cultivation copper coin tree species of secondary disinfection treatment rinses three to five times and obtains inoculation copper coin tree species, inoculation with copper coin tree species
Son is embedded in germination culture medium and is cultivated, and until growing up to seedling, cultivation temperature is 26 ± 2 DEG C, cultivates 20 to 30 days,
The culture medium that germinates is MS minimal medium,
Two, stem section micro cuttage: the seedling for growing 20 to 30 days is cut into the stem section of one to two section of band, by the base of stem section
Portion is downwardly into micro cuttage culture medium and is cultivated, and obtains middle stem section until growing lateral bud, micro cuttage culture medium is MS+1
~2mg/L 6-BA+0.1~0.4mg/L IBA+30g/L sucrose, cultivation temperature are 26 ± 2 DEG C,
Three, stem section is taken root: cut and grow to the middle stem section of 2-3cm and be inserted on root media, first dark culture three to
Six days, then carry out illumination cultivation 20 to 30 days, so that stem section is taken root can be obtained copper coin tree species seedling, root media 1/
4MS+0.5~1.5mg/L IBA+30g/L sucrose, cultivation temperature are 26 ± 2 DEG C.
The present invention devises,
MS (Murashige and Skoog) culture medium composition:
1/4MS minimal medium composition:
2 plant growth regulating substances: 6- benzylaminopurine (6-BA), indolebutyric acid (IBA).
3 carbon sources: sucrose
4 deionized waters
5 agar powders.
The present invention devises,
It the steps include:
The acquisition of aseptic seedling: copper coin tree species is taken out, with 10%KOH solution to kind of a skin under conditions of room temperature (25 DEG C)
Sofening treatment is carried out, seed handles 15min with 5% liquor natrii hypochloritis later, carries out at seed soaking after aseptic water washing 3-5 times
It manages and selects water and receive the seed that expands and carry out next step operation, 70% alcohol disinfecting 30s of the seed picked out, then with 5%
Liquor natrii hypochloritis sterilizes 10min, and aseptic water washing 3-5 times spare, and seed is inoculated into germination culture medium and is cultivated after disinfection
To seedling is grown up to, cultivation temperature is 26 ± 2 DEG C, is cultivated 20-30 days.The germination culture medium is MS minimal medium,
Stem section micro cuttage: being cut into the stem section with 1-2 section for previous step seedling obtained, and the base portion of stem section is downward
Cultivated in insertion micro cuttage culture medium to growing lateral bud, then lateral bud is cut into the stem section that band 1-2 is saved, then by its base portion to
It is cultivated in the fresh micro cuttage culture medium of lower insertion, cultivation temperature is 26 ± 2 DEG C；The micro cuttage culture medium is trained with MS
The 6- benzyl aminoadenine and indolebutyric acid that base is minimal medium and additional various concentration are supported,
Stem section is taken root: previous step lateral bud obtained is inoculated on root media, first dark culture 3-6 days, then
It carries out illumination cultivation 20-30 days, until stem section, which is taken root, can be obtained copper coin tree species seedling；The root media is the 1/4MS of improvement
Culture medium is minimal medium and additional indolebutyric acid.
The technical effects of the invention are that: copper coin tree micro cuttage technical operation is simple, and growth rate is fast, is able to solve utilization
The problem that germination percentage is low, reproduction speed is slow in copper coin tree seminal propagation seedling realizes the long-term extensive quick of copper coin tree species seedling
Proliferation can a large amount of, quickly, persistently obtain the copper coin tree test tube seedling of high quality.
In the technical scheme, micro cuttage culture medium and root media are important technical characteristic, fast based on micro cuttage
In the copper coin tree seedling fostering method of speed breeding and the technical field of culture medium, there is novelty, creativeness and practicability, at this
Term in technical solution is all that can be explained and be understood with patent document in the art.
Four, Detailed description of the invention
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below
There is attached drawing needed in technical description to be briefly described, it should be apparent that, the accompanying drawings in the following description is only this
Some embodiments of invention for those of ordinary skill in the art without creative efforts, can be with
It obtains other drawings based on these drawings.
Fig. 1 is the effect picture of copper coin tree species sprouting of the invention, micro cuttage and transplanting situation,
A: copper coin tree species is sprouted；B: growth of seedling；C: lateral bud is sprouted；D: plant after transplanting.
Five, specific embodiment
According to guidelines for examination, such as " having ", "comprising" and " comprising " term used in the present invention should understand that
For the presence or addition for not allotting one or more of the other element or combinations thereof.
In the description of the present invention, it should be noted that term " center ", "upper", "lower", "left", "right", "vertical",
The orientation or positional relationship of the instructions such as "horizontal", "inner", "outside" be based on the orientation or positional relationship shown in the drawings, merely to
Convenient for description the present invention and simplify description, rather than the device or element of indication or suggestion meaning must have a particular orientation,
It is constructed and operated in a specific orientation, therefore is not considered as limiting the invention.In addition, term " first ", " second ",
" third " is used for descriptive purposes only and cannot be understood as indicating or suggesting relative importance.
In the description of the present invention, it should be noted that unless otherwise clearly defined and limited, term " installation ", " phase
Even ", " connection " shall be understood in a broad sense, for example, it may be being fixedly connected, may be a detachable connection, or be integrally connected；It can
To be mechanical connection, it is also possible to be electrically connected；It can be directly connected, can also can be indirectly connected through an intermediary
Connection inside two elements.For the ordinary skill in the art, above-mentioned term can be understood at this with concrete condition
Concrete meaning in invention.
As long as in addition, the non-structure each other of technical characteristic involved in invention described below different embodiments
It can be combined with each other at conflict.
Below with reference to embodiment, the present invention is further described, following embodiment is intended to illustrate invention rather than to this
Invention further limits.
A kind of copper coin tree seedling fostering method quickly bred based on micro cuttage, one of one embodiment of the invention,
It the steps include:
Condition of culture: all culture mediums in cultural aim adjust pH value to 5.8 before sterilization, 1 big then at 121 DEG C
High pressure sterilization 20min under air pressure.Culturing room's cultivation temperature is 26 ± 2 DEG C, and periodicity of illumination is 16 hours/day,
One, the acquisition of aseptic seedling: being put into copper coin tree species in 10%KOH solution, the room temperature item for being 25 ± 2 DEG C in temperature
Sofening treatment is carried out to the kind skin of copper coin tree species under part, the soaking time in 10%KOH solution is set as 18-22min, soft
The copper coin tree species son distilled water flushing for changing processing is clean, then copper coin tree species of sofening treatment is put into 5% hypochlorous acid
Sodium solution carries out immersion treatment 8-12min, and sterile water is issued to after aseptic water washing three to five times and carries out seed soaking, when seed soaking
Between be set as 24-36h, and selecting copper coin tree species that expands of water suction is cultivation copper coin tree species, cultivation with copper coin tree species
Son is put into the first time disinfection treatment for carrying out that the time is 28-32s in 70% alcohol, the cultivation copper coin of first time disinfection treatment
Tree species is put into second of disinfection treatment for carrying out that the time is 8-12min in 5% liquor natrii hypochloritis, with sterile water to second
Cultivation copper coin tree species of secondary disinfection treatment rinses three to five times and obtains inoculation copper coin tree species, inoculation with copper coin tree species
Son is embedded in germination culture medium and is cultivated, and until growing up to seedling, cultivation temperature is 26 ± 2 DEG C, cultivates 20 to 30 days,
The culture medium that germinates is MS minimal medium,
Two, stem section micro cuttage: the seedling for growing 20 to 30 days is cut into the stem section of one to two section of band, by the base of stem section
Portion is downwardly into micro cuttage culture medium and is cultivated, and obtains middle stem section until growing lateral bud, micro cuttage culture medium is MS+1
~2mg/L 6-BA+0.1~0.4mg/L IBA+30g/L sucrose, cultivation temperature are 26 ± 2 DEG C,
Three, stem section is taken root: cut and grow to the middle stem section of 2-3cm and be inserted on root media, first dark culture three to
Six days, then carry out illumination cultivation 20 to 30 days, so that stem section is taken root can be obtained copper coin tree species seedling, root media 1/
4MS+0.5~1.5mg/L IBA+30g/L sucrose, cultivation temperature are 26 ± 2 DEG C.
In the present embodiment,
MS (Murashige and Skoog) culture medium composition:
1/4MS minimal medium composition:
2 plant growth regulating substances: 6- benzylaminopurine (6-BA), indolebutyric acid (IBA).
3 carbon sources: sucrose
4 deionized waters
5 agar powders.
The two of one embodiment of the invention, the steps include:
Condition of culture: all culture mediums in cultural aim adjust pH value to 5.8 before sterilization, 1 big then at 121 DEG C
High pressure sterilization 20min under air pressure.Culturing room's cultivation temperature is 26 ± 2 DEG C, and periodicity of illumination is 16 hours/day,
One, the acquisition of aseptic seedling: being put into copper coin tree species in 10%KOH solution, the room temperature condition for being 23 DEG C in temperature
Under sofening treatment carried out to the kind skin of copper coin tree species, the soaking time in 10%KOH solution is set as 18min, at softening
The copper coin tree species son distilled water flushing of reason is clean, and the sodium hypochlorite that copper coin tree species of sofening treatment is then put into 5% is molten
Liquid carries out immersion treatment 8min, and aseptic water washing is issued to sterile water afterwards three times and carries out seed soaking, and seed soaking time is set as
For 24 hours, and copper coin tree species that water suction is expanded is selected as cultivation copper coin tree species, cultivation is put into 70% wine with copper coin tree species
The first time disinfection treatment that the time is 28s is carried out in essence, the cultivation of disinfection treatment is put into 5% with copper coin tree species first time
Second of disinfection treatment that the time is 8min is carried out in liquor natrii hypochloritis, is used with cultivation of the sterile water to second of disinfection treatment
Copper coin tree species flushes three times to obtain inoculation copper coin tree species, inoculation with copper coin tree species be embedded in germination culture medium into
Row culture, until growing up to seedling, cultivation temperature is 24 DEG C, is cultivated 20 days, and germination culture medium is MS minimal medium,
Two, the seedling for growing 20 days: being cut into the stem section of one to two section of band by stem section micro cuttage, and the base portion of stem section is downward
It is cultivated in insertion micro cuttage culture medium, obtains middle stem section until growing lateral bud, micro cuttage culture medium is MS+1mg/L 6-
BA+0.1mg/L IBA+30g/L sucrose, cultivation temperature are 24 DEG C,
Three, stem section is taken root: it cuts and grows to the middle stem section of 2cm and be inserted on root media, first dark culture three days, then
It carries out illumination cultivation 20 days, so that stem section is taken root can be obtained copper coin tree species seedling, and root media is 1/4MS+0.5mg/L IBA
+ 30g/L sucrose, cultivation temperature are 24 DEG C.
The three of one embodiment of the invention, the steps include:
Condition of culture: all culture mediums in cultural aim adjust pH value to 5.8 before sterilization, 1 big then at 121 DEG C
High pressure sterilization 20min under air pressure.Culturing room's cultivation temperature is 26 ± 2 DEG C, and periodicity of illumination is 16 hours/day,
One, the acquisition of aseptic seedling: being put into copper coin tree species in 10%KOH solution, the room temperature condition for being 27 DEG C in temperature
Under sofening treatment carried out to the kind skin of copper coin tree species, the soaking time in 10%KOH solution is set as 22min, at softening
The copper coin tree species son distilled water flushing of reason is clean, and the sodium hypochlorite that copper coin tree species of sofening treatment is then put into 5% is molten
Liquid carries out immersion treatment 12min, and sterile water is issued to after aseptic water washing five times and carries out seed soaking, seed soaking time is set as
36h, and selecting copper coin tree species that water suction is expanded is cultivation copper coin tree species, and cultivation is put into 70% wine with copper coin tree species
The first time disinfection treatment that the time is 32s is carried out in essence, the cultivation of disinfection treatment is put into 5% with copper coin tree species first time
Second of disinfection treatment that the time is 12min, the cultivation with sterile water to second of disinfection treatment are carried out in liquor natrii hypochloritis
Five times are rinsed with copper coin tree species and obtains inoculation copper coin tree species, and inoculation is embedded in germination culture medium with copper coin tree species
It is cultivated, until growing up to seedling, cultivation temperature is 28 DEG C, is cultivated 30 days, and germination culture medium is MS minimal medium,
Two, the seedling for growing 30 days: being cut into the stem section of one to two section of band by stem section micro cuttage, and the base portion of stem section is downward
It is cultivated in insertion micro cuttage culture medium, obtains middle stem section until growing lateral bud, micro cuttage culture medium is MS+2mg/L 6-
BA+0.4mg/L IBA+30g/L sucrose, cultivation temperature are 28 DEG C,
Three, stem section is taken root: it cuts and grows to the middle stem section of 3cm and be inserted on root media, first dark culture six days, then
It carries out illumination cultivation 30 days, so that stem section is taken root can be obtained copper coin tree species seedling, and root media is 1/4MS+1.5mg/L IBA
+ 30g/L sucrose, cultivation temperature are 28 DEG C.
The four of one embodiment of the invention, the steps include:
Condition of culture: all culture mediums in cultural aim adjust pH value to 5.8 before sterilization, 1 big then at 121 DEG C
High pressure sterilization 20min under air pressure.Culturing room's cultivation temperature is 26 ± 2 DEG C, and periodicity of illumination is 16 hours/day,
One, the acquisition of aseptic seedling: being put into copper coin tree species in 10%KOH solution, the room temperature condition for being 25 DEG C in temperature
Under sofening treatment carried out to the kind skin of copper coin tree species, the soaking time in 10%KOH solution is set as 22min, at softening
The copper coin tree species son distilled water flushing of reason is clean, and the sodium hypochlorite that copper coin tree species of sofening treatment is then put into 5% is molten
Liquid carries out immersion treatment 10min, and sterile water is issued to after aseptic water washing four times and carries out seed soaking, seed soaking time is set as
30h, and selecting copper coin tree species that water suction is expanded is cultivation copper coin tree species, and cultivation is put into 70% wine with copper coin tree species
The first time disinfection treatment that the time is 30s is carried out in essence, the cultivation of disinfection treatment is put into 5% with copper coin tree species first time
Second of disinfection treatment that the time is 10min, the cultivation with sterile water to second of disinfection treatment are carried out in liquor natrii hypochloritis
It is rinsed four times to obtain inoculation copper coin tree species with copper coin tree species, inoculation is embedded in germination culture medium with copper coin tree species
It is cultivated, until growing up to seedling, cultivation temperature is 26 DEG C, is cultivated 25 days, and germination culture medium is MS minimal medium,
Two, stem section micro cuttage: the stem section for 25 days seedling will be grown being cut into one to two section of band, by the base portion of stem section to
Underthrust enters is cultivated in micro cuttage culture medium, obtains middle stem section until growing lateral bud, micro cuttage culture medium is MS+1.5mg/
L 6-BA+0.25mg/L IBA+30g/L sucrose, cultivation temperature are 26 DEG C,
Three, stem section is taken root: it cuts and grows to the middle stem section of 2.5cm and be inserted on root media, first dark culture five days,
It carries out again illumination cultivation 25 days, so that stem section is taken root can be obtained copper coin tree species seedling, root media 1/4MS+1.0mg/L
IBA+30g/L sucrose, cultivation temperature are 26 DEG C.
Second embodiment of the invention, the steps include:
The acquisition of aseptic seedling: copper coin tree species is taken out, with 10%KOH solution to kind of a skin under conditions of room temperature (25 DEG C)
Sofening treatment is carried out, seed handles 15min with 5% liquor natrii hypochloritis later, carries out at seed soaking after aseptic water washing 3-5 times
It manages and selects water and receive the seed that expands and carry out next step operation, 70% alcohol disinfecting 30s of the seed picked out, then with 5%
Liquor natrii hypochloritis sterilizes 10min, and aseptic water washing 3-5 times spare, and seed is inoculated into germination culture medium and is cultivated after disinfection
To seedling is grown up to, cultivation temperature is 26 ± 2 DEG C, is cultivated 20-30 days.The germination culture medium is MS minimal medium,
Stem section micro cuttage: being cut into the stem section with 1-2 section for previous step seedling obtained, and the base portion of stem section is downward
Cultivated in insertion micro cuttage culture medium to growing lateral bud, then lateral bud is cut into the stem section that band 1-2 is saved, then by its base portion to
It is cultivated in the fresh micro cuttage culture medium of lower insertion, cultivation temperature is 26 ± 2 DEG C；The micro cuttage culture medium is trained with MS
The 6- benzyl aminoadenine and indolebutyric acid that base is minimal medium and additional various concentration are supported,
Stem section is taken root: previous step lateral bud obtained is inoculated on root media, first dark culture 3-6 days, then
It carries out illumination cultivation 20-30 days, until stem section, which is taken root, can be obtained copper coin tree species seedling；The root media is the 1/4MS of improvement
Culture medium is minimal medium and additional indolebutyric acid.
Following will be combined with the drawings in the embodiments of the present invention, and technical solution in the embodiment of the present invention carries out clear, complete
Site preparation description, it is clear that described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.It is based on
Embodiment in the present invention, it is obtained by those of ordinary skill in the art without making creative efforts every other
Embodiment shall fall within the protection scope of the present invention.
A kind of copper coin tree species seedling culture medium quickly bred based on micro cuttage, one of one embodiment of the invention, packet
Contain germination culture medium, micro cuttage culture medium and root media.
In the present embodiment, germination culture medium be set as MS minimal medium, micro cuttage culture medium be set as MS+1~
2mg/L 6-BA+0.1~0.4mg/L IBA+30g/L sucrose, root media are set as 1/4MS+0.5~1.5mg/L IBA+
30g/L sucrose.
In the present embodiment,
MS (Murashige and Skoog) culture medium composition:
1/4MS minimal medium composition:
2 plant growth regulating substances: 6- benzylaminopurine (6-BA), indolebutyric acid (IBA).
3 carbon sources: sucrose
4 deionized waters
5 agar powders.
The two of one embodiment of the invention, culture medium is set as MS minimal medium, micro cuttage culture medium is set for germination
It is set to MS+1mg/L 6-BA+0.1mg/L IBA+30g/L sucrose, root media is set as 1/4MS+0.5mg/L IBA+
30g/L sucrose.
The three of one embodiment of the invention, culture medium is set as MS minimal medium, micro cuttage culture medium is set for germination
It is set to MS+2mg/L 6-BA+0.4mg/L IBA+30g/L sucrose, root media is set as 1/4MS+1.5mg/L IBA+
30g/L sucrose.
The four of one embodiment of the invention, culture medium is set as MS minimal medium, micro cuttage culture medium is set for germination
It is set to MS+1.5mg/L 6-BA+0.25mg/L IBA+30g/L sucrose, root media is set as 1/4MS+1.0mg/L IBA+
30g/L sucrose.
Verification test result of the invention:
The acquisition of aseptic seedling:
This technology carries out sofening treatment to copper coin tree species skin using 10%KOH solution, and solution copper coin tree species skin is harder,
The problem of being not easy water swelling；After carrying out initial sterilization to seed using 5% liquor natrii hypochloritis, reused after seed soaking
70% alcohol and 5% liquor natrii hypochloritis carry out secondary sterilization, then inoculation and germination culture medium, the side of this gradation disinfection
Formula can effectively control pollution rate, within Contamination rate control again 5%；And selected after soaking seed obviously absorb the seed that expands into
Row inoculation, can effectively improve the validity of operation, improve the germination rate of copper coin tree species of inoculation.Fig. 1-A is copper coin tree
Seed sprouting situation；The growing state of Fig. 1-B copper coin tree seedling.
Stem section micro cuttage:
The seedling for growing 20-30 days is cut into the stem section with 1-2 section, the base portion of stem section is downwardly into micro cuttage culture medium
In cultivated, one week or so, lateral bud started to grow, in culture medium MS+2mg/L 6-BA+0.1~0.2mg/L IBA+30g/L
On sucrose, lateral bud well-grown, stem elongation is obvious, and blade is sufficiently spread out (Fig. 1-C).
Stem section is taken root:
It takes and grows to being seeded on root media of 2-3cm lateral bud, first dark culture 3-6 days, then illumination cultivation is carried out,
Situation of taking root on the culture medium of culture medium 1/4MS+1.0mg/L IBA+30g/L sucrose is preferable, can get copper of taking root within 30 days or so
Money tree species seedling.It uncaps and is transplanted after taming 5-7 days, growing state is good (Fig. 1-D) after transplanting.
The present invention has the following characteristics:
1, due to devising micro cuttage culture medium and root media, by micro cuttage culture medium and root media, make
The parent of seedling carries out segmentation fast breeding, no longer carries out direct nursery by copper coin tree species, thus realize it is a large amount of, quickly,
Persistently obtain the copper coin tree test tube seedling of high quality.
2, due to devising germination culture medium, by the culture medium that germinates, the parent of the seedling for cultivation is obtained,.
3, it is quickly bred due to devising micro cuttage, overcomes the low technology barrier of the germination of copper coin tree, realize
Fast breeding.
4, the restriction of numberical range has been carried out to planform due to devising, has made the technical side that numberical range is of the invention
Technical characteristic in case is not to be calculated by formula or test the technical characteristic obtained by limited times, experiments have shown that the numerical value
The technical characteristic of range achieves good technical effect.
5, due to devising technical characteristic of the invention, in the effect of the independent and mutual set of technical characteristic,
By experiments have shown that, performance indexes of the invention be existing performance indexes be at least 1.7 times, pass through assessment tool
There is good market value.
It is all the present invention there are also other technical characteristics identical or similar with micro cuttage culture medium and root media
One of embodiment, and each technical characteristic of embodiment described above can be combined arbitrarily, to meet Patent Law, specially
The requirement of sharp detailed rules for the implementation and guidelines for examination, no longer to all possible combined reality of each technical characteristic in above-described embodiment
Example is applied all to be described.
Above-described embodiment be the copper coin tree seedling fostering method provided by the present invention quickly bred based on micro cuttage and
A kind of way of realization of culture medium, other deformations of provided scheme, increase or reduce composition therein according to the present invention
Or step, or the present invention is used for other technical fields close with the present invention, it all belongs to the scope of protection of the present invention.
Claims (8)
1. a kind of copper coin tree species seedling culture medium quickly bred based on micro cuttage, it is characterized in that: including for seedling parent
Stem section grows the micro cuttage culture medium of lateral bud and the root media for growing root system for the stem section of seedling parent.
2. the copper coin tree species culture medium according to claim 1 quickly bred based on micro cuttage, it is characterized in that: also including
For cultivating the germination culture medium of seedling parent.
3. the copper coin tree species seedling culture medium according to claim 2 quickly bred based on micro cuttage, it is characterized in that: germination training
Feeding base is set as MS minimal medium, micro cuttage culture medium is set as MS+1~2mg/L 6-BA+0.1~0.4mg/L IBA+
30g/L sucrose, root media are set as 1/4MS+0.5~1.5mg/L IBA+30g/L sucrose.
4. the copper coin tree species seedling culture medium according to claim 3 quickly bred based on micro cuttage, it is characterized in that:
MS (Murashige and Skoog) culture medium composition:
1/4MS minimal medium composition:
2 plant growth regulating substances: 6- benzylaminopurine (6-BA), indolebutyric acid (IBA).
3 carbon sources: sucrose
4 deionized waters
5 agar powders.
5. a kind of copper coin tree seedling fostering method quickly bred based on micro cuttage, it is characterized in that: the steps include: one to two section
Copper coin tree seedling stem section carried out in micro cuttage culture medium promote bud growth, obtain middle stem section, middle stem section taken root
Promoting root growth growth is carried out in culture medium, obtains copper coin tree species seedling.
6. the copper coin tree seedling fostering method according to claim 1 quickly bred based on micro cuttage, it is characterized in that: it is walked
Suddenly it is:
Condition of culture: all culture mediums in cultural aim adjust pH value to 5.8 before sterilization, then at 121 DEG C, 1 atmospheric pressure
Lower high pressure sterilization 20min.Culturing room's cultivation temperature is 26 ± 2 DEG C, and periodicity of illumination is 16 hours/day,
One, the acquisition of aseptic seedling: being put into copper coin tree species in 10%KOH solution, temperature be 25 ± 2 DEG C under room temperature
Sofening treatment carried out to the kind skin of copper coin tree species, the soaking time in 10%KOH solution is set as 18-22min, at softening
The copper coin tree species son distilled water flushing of reason is clean, and the sodium hypochlorite that copper coin tree species of sofening treatment is then put into 5% is molten
Liquid carries out immersion treatment 8-12min, and sterile water is issued to after aseptic water washing three to five times and carries out seed soaking, seed soaking time is set
It is set to 24-36h, and selecting copper coin tree species that water suction is expanded is cultivation copper coin tree species, and cultivation is put with copper coin tree species
Carrying out the first time that the time is 28-32s into 70% alcohol disinfects, the cultivation of first time disinfection treatment with copper coin tree species
It is second of 8-12min disinfection treatment that son, which is put into 5% liquor natrii hypochloritis and carries out the time, with sterile water to disappearing for the second time
Cultivation copper coin tree species of poison processing rinses three to five times and obtains inoculation copper coin tree species, and inoculation is buried with copper coin tree species
Enter into germination culture medium and cultivated, until growing up to seedling, cultivation temperature is 26 ± 2 DEG C, is cultivated 20 to 30 days, germination
Culture medium is MS minimal medium,
Two, stem section micro cuttage: the stem section for 20 to 30 days seedling will be grown being cut into one to two section of band, by the base portion of stem section to
Underthrust enters is cultivated in micro cuttage culture medium, obtains middle stem section until growing lateral bud, micro cuttage culture medium be MS+1~
2mg/L 6-BA+0.1~0.4mg/L IBA+30g/L sucrose, cultivation temperature are 26 ± 2 DEG C,
Three, stem section is taken root: it cuts and grows to the middle stem section of 2-3cm and be inserted on root media, first dark culture three to six days,
Illumination cultivation is carried out again 20 to 30 days, so that stem section is taken root can be obtained copper coin tree species seedling, root media 1/4MS+0.5
~1.5mg/L IBA+30g/L sucrose, cultivation temperature are 26 ± 2 DEG C.
7. the copper coin tree seedling fostering method according to claim 6 quickly bred based on micro cuttage, it is characterized in that:
MS (Murashige and Skoog) culture medium composition:
1/4MS minimal medium composition:
2 plant growth regulating substances: 6- benzylaminopurine (6-BA), indolebutyric acid (IBA).
3 carbon sources: sucrose
4 deionized waters
5 agar powders.
8. the copper coin tree seedling fostering method according to claim 5 quickly bred based on micro cuttage, it is characterized in that:
It the steps include:
The acquisition of aseptic seedling: copper coin tree species is taken out, kind of skin is carried out under conditions of room temperature (25 DEG C) with 10%KOH solution
Sofening treatment, seed handles 15min with 5% liquor natrii hypochloritis later, carries out seed soaking simultaneously after aseptic water washing 3-5 times
It selects water and receives the seed that expands and carry out next step operation, 70% alcohol disinfecting 30s of the seed picked out, then with 5% secondary chlorine
Acid sodium solution sterilizes 10min, and aseptic water washing 3-5 times spare, and seed is inoculated into germination culture medium and is cultivated to length after disinfection
At seedling, cultivation temperature is 26 ± 2 DEG C, is cultivated 20-30 days.The germination culture medium is MS minimal medium,
Stem section micro cuttage: previous step seedling obtained is cut into the stem section with 1-2 section, the base portion of stem section is downwardly into
It is cultivated in micro cuttage culture medium to growing lateral bud, then lateral bud is cut into the stem section with 1-2 section, then by its base portion to underthrust
Enter and cultivated in fresh micro cuttage culture medium, cultivation temperature is 26 ± 2 DEG C；The micro cuttage culture medium is with MS culture medium
For minimal medium and the 6- benzyl aminoadenine and indolebutyric acid of additional various concentration,
Stem section is taken root: previous step lateral bud obtained being inoculated on root media, first dark culture 3-6 days, then carried out
Illumination cultivation 20-30 days, until stem section, which is taken root, can be obtained copper coin tree species seedling；The root media is the 1/4MS culture of improvement
Base is minimal medium and additional indolebutyric acid.
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| CN105532121A (en) * | 2016-01-09 | 2016-05-04 | 黄庆梅 | Sterile sowing method for ginseng seeds |
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| JPS57189605A (en) * | 1981-05-16 | 1982-11-22 | Kiyuuzou Kamata | Production of agar like growing soil |
| CN1263949A (en) * | 2000-01-26 | 2000-08-23 | 山东大学 | Method for creating transgenic receptor system of corn and application of same |
| CN1460713A (en) * | 2003-04-29 | 2003-12-10 | 广西富新科技股份有限公司 | Camellia chrysantha culture medium for tissue culture and quick breeding |
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