CN107347643A - A kind of U.S.'s gold leaf Chinese honey locust tissue culture and rapid propagation method - Google Patents
A kind of U.S.'s gold leaf Chinese honey locust tissue culture and rapid propagation method Download PDFInfo
- Publication number
- CN107347643A CN107347643A CN201710651830.4A CN201710651830A CN107347643A CN 107347643 A CN107347643 A CN 107347643A CN 201710651830 A CN201710651830 A CN 201710651830A CN 107347643 A CN107347643 A CN 107347643A
- Authority
- CN
- China
- Prior art keywords
- explant
- culture
- honey locust
- chinese honey
- gold leaf
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
It is of the invention that a kind of U.S.'s gold leaf Chinese honey locust tissue culture and rapid propagation method is provided, including choose explant, explant sterilization, pretreatment, Primary culture, differentiation culture, cultivation tissue-cultured seedling, hardening and domestication step；Described selection explant：Explant chooses the terminal bud stem section of the vigorous branch of tree crown top peripheral, it is desirable to which terminal bud is full, and stem section length is 1 1.8cm；With scissors clip explant, environment temperature is 22 28 DEG C during clip explant, and humidity is 65 70%.Described pretreatment：By gibberellin, 6 BA, IBA, potassium dihydrogen phosphate according to 2:1:2:The 0.05 mg/L aqueous solution is configured to after 3 ratio mixing, obtains pretreatment fluid；Then the explant after sterilization is put into pretreatment fluid and soaked 8 hours.Using gold leaf Chinese honey locust tissue culture and rapid propagation method in the U.S.'s of the present invention, not only reproduction speed is fast, and survival rate of plant improves 15.5%.
Description
Technical field
The present invention relates to a kind of U.S.'s gold leaf Chinese honey locust mating system, and in particular to a kind of U.S.'s gold leaf Chinese honey locust tissue-culturing rapid propagation side
Method, belong to breeding plant technical field.
Background technology
U.S. gold leaf Chinese honey locust, broadleaf deciduous arbor, 9~10.5 meters of plant height, growth is very fast, and no scolus, spire is golden yellow,
Gold leaf Chinese honey locust climax leaves chartreuse, still it is chartreuse to autumn, branch is unfolded, and plant type is beautiful, shaky.
Property light and slightly it is resistance to it is shady, drought-enduring, cold-resistant, be resistant to -34 degree low temperature.Like warm and moist weather and deep fertile soil
Earth, it can also be grown in calcium carbonate and slight saline-alkali soil, adaptability is wider, and NORTH CHINA to south and the west and south can plant
Plant.Deep-rootedness, few pest and disease damage.Found in production, easy torticollis in below diameter of a cross-section of a tree trunk 1.3 meters above the ground 3cm plant strain growth.
Northern area is due to very cold, and plant variety is single, and color leafed plants lack, and arbor type Colored- leaf Plants especially lack
It is weary, gold leaf Chinese honey locust it is leaf it is beautiful, leaf color is golden yellow, its successful popularization, this regional landscape effect will be greatly enriched.Current
U.S.'s gold leaf Chinese honey locust raising technology is mostly grafting, but very fast due to growing, and is easily snapped off at grafting, resistance is not strong；And tissue culture
The U.S. gold leaf Chinese honey locust of breeding not only overcomes drawbacks described above, and breeds quickly, and seedling strain is virus-free, and diseases and insect pests resistance is strong.
The report of U.S.'s gold leaf Chinese honey locust group culturation rapid propagating technology is now there is no in the art.
The content of the invention
In view of the shortcomings of the prior art, it is numerous it is an object of the invention to provide a kind of U.S.'s gold leaf Chinese honey locust tissue culture and rapid propagation method
It is fast to grow speed, survival rate is high, and the U.S.'s gold leaf Chinese honey locust diseases and insect pests resistance cultivated is strong, and plant is not susceptible to torticollis.
To solve existing technical problem, the technical solution adopted by the present invention is：
A kind of U.S.'s gold leaf Chinese honey locust tissue culture and rapid propagation method, including choose explant step；Described selection explant：Explant selects
Take the terminal bud stem section of the vigorous branch of tree crown top peripheral, it is desirable to which terminal bud is full, and stem section length is 1~1.8cm；With scissors clip
Explant, environment temperature is 22~28 DEG C during clip explant, and humidity is 65~70%.
Described method also includes explant sterilisation step；Described explant sterilization：（1）Explant is placed in triangular flask
It is interior, 75% 1~2min of alcohol disinfecting is added, then rinsed repeatedly with clear water 3 times, rinse well；By explant with 0.1% mercuric chloride
Sterilize 5~10min；Aseptic water washing 8 times.
Described method also includes pre-treatment step；Described pretreatment：By gibberellin, 6-BA, IBA, potassium dihydrogen phosphate
According to 2:1:2:The 0.05 mg/L aqueous solution is configured to after 3 ratio mixing, obtains pretreatment fluid；Then will be outer after sterilization
Implant, which is put into pretreatment fluid, soaks 8 hours.
Described method also includes Primary culture step；Described Primary culture：
The condition of culture is controlled to be：28 ± 0.5 DEG C of environment temperature is controlled, light application time is 5 ± 0.5 h/d, intensity of illumination 2700
~3000 Lx, humidity are 60~65%.
Described method also includes differentiation incubation step, described differentiation culture：
The differential medium used for：The mg/L+ honey 8g/L+ agar 9 of MS culture medium+NAA 0.15mg/L+IAA 0.05
G/L, regulation pH are 6.5.
Described method also includes cultivating tissue-cultured seedling step, described cultivation tissue-cultured seedling：
The Multiple Buds that differentiation culture obtains are inoculated on strengthening seedling and rooting culture medium, described strengthening seedling and rooting culture medium is that MS is cultivated
Base+6-BA 0.08mg/L+ NAA 0.12mg/L+pollen pini 0.25mg/L+folic acid 0.1mg/L+g/L of agar 9, regulation
PH is 6.5.
Described cultivation tissue-cultured seedling：
By be inoculated with Multiple Buds culture medium be placed in light application time be 10h/d, intensity of illumination 3300Lx, culture room temperature be 28 DEG C
Under conditions of cultivate 25~30 days, obtain tissue-cultured seedling.
Described method also includes hardening step, described hardening：
By tissue culture transplantation of seedlings in culture matrix, overlay film, intensity of illumination be 3200 Lx under conditions of, control temperature be 28~
30 DEG C, humidity is 65~70%, hardening culture 33~35 days, obtains the good seedling of root system development.
Described culture matrix is by vermiculite, perlite, jute, cinnamomi cortex pulveratus, graphene, turf according to 10：7：5：1：4：3
Mass ratio mixes, and sprays into water, and it is 45% to control water content.
Described method also includes domestication step；Described domestication：
It is 65~70% to control ambient humidity, and intensity of illumination is 3100~3300Lx, and light application time is 8~10h/d, air flow
Measure as 100~120m3/h；Using airconditioning control environment temperature：Day temperature is warming up to 28 DEG C from 18 DEG C of speed with 2 DEG C/h,
Being kept for 28 DEG C two hours, then 18 DEG C are cooled to 2 DEG C/h speed, nocturnal temperature is cooled to 12 DEG C with 1 DEG C/h speed, then with
1 DEG C/h speed is warming up to 18 DEG C；Continuous domestication 21~25 days.
By adopting the above-described technical solution, the present invention reach have the technical effect that：
（1）Using gold leaf Chinese honey locust tissue culture and rapid propagation method in the U.S.'s of the present invention, survival rate of plant improves 15.5%, up to 99.5%；
（2）Using gold leaf Chinese honey locust tissue culture and rapid propagation method in the U.S.'s of the present invention, plant strain growth speed is fast, transplants plant height then
Increase by 98~105cm；
（3）The U.S.'s gold leaf Chinese honey locust resistance against diseases cultivated using U.S.'s gold leaf Chinese honey locust tissue culture and rapid propagation method of the present invention is strong, is not easy to send out
Raw Disease, diameter of a cross-section of a tree trunk 1.3 meters above the ground 3cm plant occur without torticollis symptom；The insect pest incidence lower than common plant 12.5% of longicorn；It is resistant to
Subzero 40 DEG C of low temperature.
Embodiment
A kind of U.S.'s gold leaf Chinese honey locust tissue culture and rapid propagation method of embodiment 1
1st, material prepares
Test and carried out in 2015 at Weifang Vocatonal College's tissue culture center, test material is U.S. gold leaf Chinese honey locust（Gleditsla
triacanthos‘Sunburst’）.
2nd, explant is chosen
Vigorous, no disease and pests harm, the excellent U.S.'s gold leaf Chinese honey locust individual plant of trait expression are grown in outdoor selection；With scissors clip explant
Body, environment temperature is 25 DEG C during clip explant, humidity 66%；
The explant of clip is：The terminal bud stem section of the vigorous branch of tree crown top peripheral, it is desirable to which terminal bud is full, and stem section length is
1.0cm。
3rd, explant sterilizes
Explant is placed in triangular flask, adds 75% alcohol disinfecting 1min, then is rinsed repeatedly with clear water 3 times, is rinsed well；
In superclean bench, explant is sterilized into 7min with 0.1% mercuric chloride；Aseptic water washing 8 times, it is standby.
4th, pre-process
By gibberellin, 6-BA, IBA, potassium dihydrogen phosphate according to 2:1:2:The water-soluble of 0.05 mg/L is configured to after 3 ratio mixing
Liquid, obtain pretreatment fluid；Then the explant after sterilization is put into pretreatment fluid and soaked 8 hours.
5th, Primary culture
The primary culture medium used for：
With distilled water primary culture medium is prepared according to formula as below：
NH4NO3 1.650 g/L、KNO3 1.900 g/L、CaCl2·2H2O 440 mg/L、MgSO4·7H2O 370 mg/L、
KH2PO4 700 mg/L、KI 0.83 mg/L、H3BO3 6.2 mg/L、MnSO4·4H2O 22.3 mg/L、ZnSO4·7H2O
8.6 mg/L、Na2MnO4·2H2O 0.25 mg/L、CuSO4·5H2O 0.025 mg/L、CoCl2·6H2O 0.025 mg/L、
FeSO4·7H2O 27.8 mg/L、Na2-EDTA·2H2The mg/L of O 37.3, inositol 85mg/L, the mg/L of nicotinic acid 1.1, hydrochloric acid
The mg/L of pyridoxol 0.2, the mg/L of thiamine hydrochloride 0.8, the mg/L of aspartic acid 1.2mg/L, 6-BA 0.05, heteroauxin
0.08 mg/L, the g/L of chlorogenic acid 0.85, the g/L of agar 9.
The primary culture medium adjustment pH prepared is 6.8；By high pressure steam sterilization, and 25 DEG C are cooled to, it is standby.
Pretreated explant is put into primary culture medium, Primary culture；The condition of culture is controlled to be：Control environment temperature
28 ± 0.5 DEG C of degree, light application time are 5 h/d, and intensity of illumination is 2700 Lx, humidity 60%.
Incubation time is 20 days, and terminal bud is scaled off from bastem portion, standby.
6th, differentiation culture
Terminal bud is inoculated on differential medium, differential medium is：The mg/ of MS culture medium+NAA 0.15mg/L+IAA 0.05
The g/L of L+ honey 8g/L+ agar 9, regulation pH are 6.5；
Be placed in light application time be 11h/d, intensity of illumination 3100Lx, culture room temperature be 27 DEG C under conditions of cultivate 28 days, obtain
It is to a large amount of Multiple Buds, i.e. adventitious bud, single Multiple Buds are scattered out, it is standby.
7th, tissue-cultured seedling is cultivated
Multiple Buds are inoculated on strengthening seedling and rooting culture medium, described strengthening seedling and rooting culture medium is MS culture mediums+6-BA
The mg/L+ pollen pinis 0.25mg/L of 0.08mg/L+ NAA 0.12mg/L+cod-liver oil 0.02+folic acid 0.1mg/L+agar 9
G/L, regulation pH are 6.5；
Be placed in light application time be 10h/d, intensity of illumination 3300Lx, culture room temperature be 28 DEG C under conditions of cultivate 25 days, obtain
To tissue-cultured seedling.
8th, hardening
By tissue culture transplantation of seedlings in culture matrix, overlay film, under conditions of intensity of illumination is 3200 Lx, it is 28 DEG C to control temperature,
Humidity is 65%, hardening culture 33 days, obtains the good seedling of root system development；
Described culture matrix is by vermiculite, perlite, jute, cinnamomi cortex pulveratus, graphene, turf according to 10：7：5：1：4：3 quality
Than mixing, water is sprayed into, it is 45% to control water content.
9th, tame
By seedling replanting into greenhouse, during transplanting, 0.4m seedling cave is dug, 2g root-inducing powders are sprinkled into seedling cave and are mixed with soil, are put into children
Seedling, earthing cover seedlings root, pour once permeable, and it is 65%, intensity of illumination 3300Lx to control ambient humidity, and light application time is
10h/d, air flow amount 120m3/h；
Using airconditioning control environment temperature：Day temperature is warming up to 28 DEG C from 18 DEG C of speed with 2 DEG C/h, keeps 28 DEG C two small
When, then 18 DEG C are cooled to 2 DEG C/h speed, nocturnal temperature is cooled to 12 DEG C, then the speed with 1 DEG C/h with 1 DEG C/h speed
It is warming up to 18 DEG C；
Continuous domestication 20~22 days.
Influence experiment of the explant specification that embodiment 2 is chosen to survival rate
Method according to embodiment 1 carries out tissue culture propagation, only changes the explant specification chosen in step 2, i.e. terminal bud stem section is grown
Degree, embodiment 2~4 is carried out, is specifically shown in Table 1：
The explant specification that table 1 is chosen
The tissue-cultured seedling of investigation embodiment 1~4 survives situation；
Investigation result is shown in Table 2；
The tissue culture shoot survival percent of table 2
Investigation result is shown, the tissue culture shoot survival percent highest of embodiment 3, is preferred embodiment；The explant chosen in step 2
Specification, terminal bud stem section length is preferably 1.8cm.
Influence of the different strengthening seedling and rooting culture mediums of embodiment 5 to tissue-cultured seedling
Method according to embodiment 1 carries out tissue culture propagation, only the strengthening seedling and rooting culture medium in change step 7, and progress embodiment 5~
8：
The strengthening seedling and rooting culture medium that embodiment 5 uses for：
MS culture medium+6-BA 0.08mg/L+ NAA 0.12mg/L+mg/L+ pollen pinis the 0.25mg/L of cod-liver oil 0.02+fine jade
The g/L of fat 9；
The strengthening seedling and rooting culture medium that embodiment 6 uses for：
MS culture medium+6-BA 0.08mg/L+ NAA 0.12mg/L+pollen pini 0.25mg/L+folic acid 0.1mg/L+agar 9
g/L；
The strengthening seedling and rooting culture medium that embodiment 7 uses for：
MS culture medium+6-BA 0.08mg/L+ IBA 0.12mg/L+mg/L+ pollen pinis the 0.25mg/L of cod-liver oil 0.02+fine jade
The g/L of fat 9；
The strengthening seedling and rooting culture medium that embodiment 8 uses for：
MS culture medium+6-BA 0.08mg/L+ NAA 0.12mg/L+0.02 mg/L of cod-liver oil+folic acid 0.1mg/L+agar
9 g/L；
The tissue-cultured seedling growing state statistical result that embodiment 5~8 is cultivated is shown in Table 3；
The tissue-cultured seedling growing state of table 3
As seen from the above table, the tissue-cultured seedling growth of embodiment 6 is obvious very fast, is preferred embodiment, i.e. strengthening seedling and rooting culture medium is preferred
For：MS culture medium+6-BA 0.08mg/L+ NAA 0.12mg/L+pollen pini 0.25mg/L+folic acid 0.1mg/L+agar 9
g/L。
A kind of U.S.'s gold leaf Chinese honey locust tissue culture and rapid propagation method of embodiment 9
1st, material prepares
Test and carried out in 2015 at Weifang Vocatonal College's tissue culture center, test material is U.S. gold leaf Chinese honey locust（Gleditsla
triacanthos‘Sunburst’）.
2nd, explant is chosen
Vigorous, no disease and pests harm, the excellent U.S.'s gold leaf Chinese honey locust individual plant of trait expression are grown in outdoor selection；With scissors clip explant
Body, environment temperature is 25 DEG C during clip explant, humidity 66%；
The explant of clip is：The terminal bud stem section of the vigorous branch of tree crown top peripheral, it is desirable to which terminal bud is full, and stem section length is
1.8cm。
3rd, explant sterilizes
Explant is placed in triangular flask, adds 75% alcohol disinfecting 1min, then is rinsed repeatedly with clear water 3 times, is rinsed well；
In superclean bench, explant is sterilized into 7min with 0.1% mercuric chloride；Aseptic water washing 8 times, it is standby.
4th, pre-process
By gibberellin, 6-BA, IBA, potassium dihydrogen phosphate according to 2:1:2:The water-soluble of 0.05 mg/L is configured to after 3 ratio mixing
Liquid, obtain pretreatment fluid；Then the explant after sterilization is put into pretreatment fluid and soaked 8 hours.
Through experiment, compared with the explant without pretreatment, by the group of the pretreated explant culture of the present invention
Training shoot survival percent is substantially higher by 23% or so, and the speed of growth is very fast.
5th, Primary culture
With distilled water primary culture medium is prepared according to formula as below：
NH4NO3 1.650 g/L、KNO3 1.900 g/L、CaCl2·2H2O 440 mg/L、MgSO4·7H2O 370 mg/L、
KH2PO4 700 mg/L、KI 0.83 mg/L、H3BO3 6.2 mg/L、MnSO4·4H2O 22.3 mg/L、ZnSO4·7H2O
8.6 mg/L、Na2MnO4·2H2O 0.25 mg/L、CuSO4·5H2O 0.025 mg/L、CoCl2·6H2O 0.025 mg/L、
FeSO4·7H2O 27.8 mg/L、Na2-EDTA·2H2The mg/L of O 37.3, inositol 85mg/L, the mg/L of nicotinic acid 1.1, hydrochloric acid
The mg/L of pyridoxol 0.2, the mg/L of thiamine hydrochloride 0.8, the mg/L of aspartic acid 1.2mg/L, 6-BA 0.05, heteroauxin
0.08 mg/L, the g/L of chlorogenic acid 0.85, the g/L of agar 9.
The primary culture medium adjustment pH prepared is 6.8；By high pressure steam sterilization, and 25 DEG C are cooled to, it is standby.
Pretreated explant is put into primary culture medium, Primary culture；The condition of culture is controlled to be：Control environment temperature
28 ± 0.5 DEG C of degree, light application time are 5 h/d, and intensity of illumination is 2700 Lx, humidity 60%.
Incubation time is 20 days, and terminal bud is scaled off from bastem portion, standby.
6th, differentiation culture
Terminal bud is inoculated on differential medium, differential medium is：The mg/ of MS culture medium+NAA 0.15mg/L+IAA 0.05
The g/L of L+ honey 8g/L+ agar 9, regulation pH are 6.5；
Be placed in light application time be 11h/d, intensity of illumination 3100Lx, culture room temperature be 27 DEG C under conditions of cultivate 28 days, obtain
It is to a large amount of Multiple Buds, i.e. adventitious bud, single Multiple Buds are scattered out, it is standby.
7th, tissue-cultured seedling is cultivated
Multiple Buds are inoculated on strengthening seedling and rooting culture medium, described strengthening seedling and rooting culture medium is MS culture mediums+6-BA
The 0.08mg/L+ NAA 0.12mg/L+pollen pini 0.25mg/L+folic acid 0.1mg/L+g/L of agar 9, regulation pH are 6.5；
Be placed in light application time be 10h/d, intensity of illumination 3300Lx, culture room temperature be 28 DEG C under conditions of cultivate 25 days, obtain
To tissue-cultured seedling.
8th, hardening
By tissue culture transplantation of seedlings in culture matrix, overlay film, under conditions of intensity of illumination is 3200 Lx, it is 28 DEG C to control temperature,
Humidity is 65%, hardening culture 33 days, obtains the good seedling of root system development；
Described culture matrix is by vermiculite, perlite, jute, cinnamomi cortex pulveratus, graphene, turf according to 10：7：5：1：4：3 quality
Than mixing, water is sprayed into, it is 45% to control water content.
9th, tame
By seedling replanting into greenhouse, during transplanting, 0.4m seedling cave is dug, 2g root-inducing powders are sprinkled into seedling cave and are mixed with soil, are put into children
Seedling, earthing cover seedlings root, pour once permeable, and it is 65%, intensity of illumination 3300Lx to control ambient humidity, and light application time is
10h/d, air flow amount 120m3/h；
Using airconditioning control environment temperature：Day temperature is warming up to 28 DEG C from 18 DEG C of speed with 2 DEG C/h, keeps 28 DEG C two small
When, then 18 DEG C are cooled to 2 DEG C/h speed, nocturnal temperature is cooled to 12 DEG C, then the speed with 1 DEG C/h with 1 DEG C/h speed
It is warming up to 18 DEG C；
Continuous domestication 21 days.
Through experiment, the tissue culture method of the present embodiment is compared to conventional engrafting method：Survival rate of plant raising 15.5%, it is reachable
99.5%；Plant strain growth speed is fast, and the plant height transplanted then increases 98-105cm；
Through survey：U.S.'s gold leaf Chinese honey locust diseases and insect pests resistance that the present embodiment is cultivated is strong, is not susceptible to Disease, chest
Footpath 3cm plant occur without torticollis symptom；The insect pest incidence lower than common plant 12.5% of longicorn；It is resistant to -40 DEG C of low temperature.Unless
Specified otherwise and unit commonly used in the art, ratio of the present invention, are mass ratio, the percentage, are quality percentage
Than.
Finally it should be noted that：The preferred embodiments of the present invention are the foregoing is only, are not intended to limit the invention,
Although the present invention is described in detail with reference to the foregoing embodiments, for those skilled in the art, it still may be used
To be modified to the technical scheme described in foregoing embodiments, or equivalent substitution is carried out to which part technical characteristic.
Within the spirit and principles of the invention, any modification, equivalent substitution and improvements made etc., it should be included in the present invention's
Within protection domain.
Claims (10)
- A kind of 1. U.S.'s gold leaf Chinese honey locust tissue culture and rapid propagation method, it is characterised in that：Including choosing explant step；Described selection explant：Explant chooses the top of the vigorous branch of tree crown top peripheral Leaf stem section, it is desirable to which terminal bud is full, and stem section length is 1-1.8cm；With scissors clip explant, environment temperature is during clip explant 22-28 DEG C, humidity 65-70%.
- A kind of 2. U.S.'s gold leaf Chinese honey locust tissue culture and rapid propagation method according to claim 1, it is characterised in that：Described method is also Including explant sterilisation step；Described explant sterilization：Explant is placed in triangular flask, add 75% alcohol disinfecting 1~ 2min, then rinsed repeatedly with clear water 3 times, rinse well；Explant is sterilized into 5~10min with 0.1% mercuric chloride；Aseptic water washing 8 times.
- A kind of 3. U.S.'s gold leaf Chinese honey locust tissue culture and rapid propagation method according to claim 1, it is characterised in that：Described method is also Including pre-treatment step；Described pretreatment：By gibberellin, 6-BA, IBA, potassium dihydrogen phosphate according to 2:1:2:3 ratio mixing The 0.05 mg/L aqueous solution is configured to afterwards, obtains pretreatment fluid；Then the explant after sterilization is put into pretreatment fluid and soaked Bubble 8 hours.
- A kind of 4. U.S.'s gold leaf Chinese honey locust tissue culture and rapid propagation method according to claim 1, it is characterised in that：Described method is also Including Primary culture step；Described Primary culture：The condition of culture is controlled to be：28 ± 0.5 DEG C of environment temperature is controlled, light application time is 5 ± 0.5 h/d, intensity of illumination 2700 ~3000 Lx, humidity are 60~65%.
- A kind of 5. U.S.'s gold leaf Chinese honey locust tissue culture and rapid propagation method according to claim 1, it is characterised in that：Described method is also Including breaking up incubation step, described differentiation culture：The differential medium used for：The mg/L+ honey 8g/L+ agar 9 of MS culture medium+NAA 0.15mg/L+IAA 0.05 G/L, regulation pH are 6.5.
- A kind of 6. U.S.'s gold leaf Chinese honey locust tissue culture and rapid propagation method according to claim 1, it is characterised in that：Described method is also Including cultivating tissue-cultured seedling step, described cultivation tissue-cultured seedling：The Multiple Buds that differentiation culture obtains are inoculated on strengthening seedling and rooting culture medium, described strengthening seedling and rooting culture medium is that MS is cultivated Base+6-BA 0.08mg/L+ NAA 0.12mg/L+pollen pini 0.25mg/L+folic acid 0.1mg/L+g/L of agar 9, regulation PH is 6.5.
- A kind of 7. U.S.'s gold leaf Chinese honey locust tissue culture and rapid propagation method according to claim 6, it is characterised in that：Described cultivation group Train seedling：By be inoculated with Multiple Buds culture medium be placed in light application time be 10h/d, intensity of illumination 3300Lx, culture room temperature be 28 DEG C Under conditions of cultivate 25~30 days, obtain tissue-cultured seedling.
- A kind of 8. U.S.'s gold leaf Chinese honey locust tissue culture and rapid propagation method according to claim 1, it is characterised in that：Described method is also Including hardening step, described hardening：By tissue culture transplantation of seedlings in culture matrix, overlay film, intensity of illumination be 3200 Lx under conditions of, control temperature be 28~ 30 DEG C, humidity is 65~70%, hardening culture 33~35 days, obtains the good seedling of root system development.
- A kind of 9. U.S.'s gold leaf Chinese honey locust tissue culture and rapid propagation method according to claim 8, it is characterised in that：Described culture medium Matter is by vermiculite, perlite, jute, cinnamomi cortex pulveratus, graphene, turf according to 10：7：5：1：4：3 mass ratio mixing, sprays into water, control Water content processed is 45%.
- A kind of 10. U.S.'s gold leaf Chinese honey locust tissue culture and rapid propagation method according to claim 1, it is characterised in that：Described method Also include domestication step；Described domestication：It is 65~70% to control ambient humidity, and intensity of illumination is 3100~3300Lx, and light application time is 8~10h/d, air flow Measure as 100~120m3/h；Using airconditioning control environment temperature：Day temperature is warming up to 28 DEG C from 18 DEG C of speed with 2 DEG C/h, Being kept for 28 DEG C two hours, then 18 DEG C are cooled to 2 DEG C/h speed, nocturnal temperature is cooled to 12 DEG C with 1 DEG C/h speed, then with 1 DEG C/h speed is warming up to 18 DEG C；Continuous domestication 21~25 days.
Priority Applications (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811556236.8A CN109430057A (en) | 2017-08-02 | 2017-08-02 | A kind of U.S.'s gold leaf Chinese honey locust tissue culture and rapid propagation method of high survival rate |
| CN201811556235.3A CN109452173A (en) | 2017-08-02 | 2017-08-02 | A kind of U.S.'s gold leaf Chinese honey locust breeding method that disease resistance is strong |
| CN201811557434.6A CN109566413A (en) | 2017-08-02 | 2017-08-02 | A kind of breeding method of low temperature resistant U.S.'s gold leaf Chinese honey locust |
| CN201710651830.4A CN107347643A (en) | 2017-08-02 | 2017-08-02 | A kind of U.S.'s gold leaf Chinese honey locust tissue culture and rapid propagation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710651830.4A CN107347643A (en) | 2017-08-02 | 2017-08-02 | A kind of U.S.'s gold leaf Chinese honey locust tissue culture and rapid propagation method |
Related Child Applications (3)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811556235.3A Division CN109452173A (en) | 2017-08-02 | 2017-08-02 | A kind of U.S.'s gold leaf Chinese honey locust breeding method that disease resistance is strong |
| CN201811557434.6A Division CN109566413A (en) | 2017-08-02 | 2017-08-02 | A kind of breeding method of low temperature resistant U.S.'s gold leaf Chinese honey locust |
| CN201811556236.8A Division CN109430057A (en) | 2017-08-02 | 2017-08-02 | A kind of U.S.'s gold leaf Chinese honey locust tissue culture and rapid propagation method of high survival rate |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107347643A true CN107347643A (en) | 2017-11-17 |
Family
ID=60287405
Family Applications (4)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811556236.8A Pending CN109430057A (en) | 2017-08-02 | 2017-08-02 | A kind of U.S.'s gold leaf Chinese honey locust tissue culture and rapid propagation method of high survival rate |
| CN201811557434.6A Pending CN109566413A (en) | 2017-08-02 | 2017-08-02 | A kind of breeding method of low temperature resistant U.S.'s gold leaf Chinese honey locust |
| CN201811556235.3A Pending CN109452173A (en) | 2017-08-02 | 2017-08-02 | A kind of U.S.'s gold leaf Chinese honey locust breeding method that disease resistance is strong |
| CN201710651830.4A Pending CN107347643A (en) | 2017-08-02 | 2017-08-02 | A kind of U.S.'s gold leaf Chinese honey locust tissue culture and rapid propagation method |
Family Applications Before (3)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811556236.8A Pending CN109430057A (en) | 2017-08-02 | 2017-08-02 | A kind of U.S.'s gold leaf Chinese honey locust tissue culture and rapid propagation method of high survival rate |
| CN201811557434.6A Pending CN109566413A (en) | 2017-08-02 | 2017-08-02 | A kind of breeding method of low temperature resistant U.S.'s gold leaf Chinese honey locust |
| CN201811556235.3A Pending CN109452173A (en) | 2017-08-02 | 2017-08-02 | A kind of U.S.'s gold leaf Chinese honey locust breeding method that disease resistance is strong |
Country Status (1)
| Country | Link |
|---|---|
| CN (4) | CN109430057A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112400695A (en) * | 2020-12-28 | 2021-02-26 | 内蒙古蒙草生态环境（集团）股份有限公司 | Culture medium for culturing evergreen common summer pink |
| CN112400696A (en) * | 2020-12-28 | 2021-02-26 | 内蒙古蒙草生态环境（集团）股份有限公司 | Tissue culture method of evergreen common selfheal fruit-spike bamboo |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115413577B (en) * | 2022-08-31 | 2023-01-24 | 河北农业大学 | Tissue culture and rapid propagation method for gleditschia horrida |
| CN116458427B (en) * | 2023-04-14 | 2024-01-26 | 广西壮族自治区农业科学院 | Method for producing root-knot potatoes in asparagus bottle |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104823852A (en) * | 2015-05-08 | 2015-08-12 | 长阳勤劳农夫农产品有限公司 | Dendrobium officinale rapid breeding method through root tip tissue culture |
| CN105766655A (en) * | 2016-04-28 | 2016-07-20 | 湖南师范大学 | Establishing method of Gleditsia vestita Chun et Howex B.G.Li.tissue culture regeneration system |
-
2017
- 2017-08-02 CN CN201811556236.8A patent/CN109430057A/en active Pending
- 2017-08-02 CN CN201811557434.6A patent/CN109566413A/en active Pending
- 2017-08-02 CN CN201811556235.3A patent/CN109452173A/en active Pending
- 2017-08-02 CN CN201710651830.4A patent/CN107347643A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104823852A (en) * | 2015-05-08 | 2015-08-12 | 长阳勤劳农夫农产品有限公司 | Dendrobium officinale rapid breeding method through root tip tissue culture |
| CN105766655A (en) * | 2016-04-28 | 2016-07-20 | 湖南师范大学 | Establishing method of Gleditsia vestita Chun et Howex B.G.Li.tissue culture regeneration system |
Non-Patent Citations (7)
| Title |
|---|
| ALI K. BASBAA等: "Multiplication vegetative in vitro de Gleditsia", 《ACTA BOTANICA GALLICA》 * |
| 兰彦平等: "培养基酸度对皂荚茎段试管培养生长的影响", 《中国农学通报》 * |
| 刘自力: "《林产化工生产技术》", 31 May 2005, 江西科学技术出版社 * |
| 江培良: "《实用本草药性歌括》", 31 January 2007, 中国医药科技出版社 * |
| 胡颂平: "《植物细胞组织培养技术》", 31 August 2014, 中国农业大学出版社 * |
| 郑晴霞等: "绒毛皂荚的组织培养", 《植物生理学通讯》 * |
| 郑晴霞等: "绒毛皂荚离体茎段培养及再生植株的细胞组织学研究", 《湖南农学院学报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112400695A (en) * | 2020-12-28 | 2021-02-26 | 内蒙古蒙草生态环境（集团）股份有限公司 | Culture medium for culturing evergreen common summer pink |
| CN112400696A (en) * | 2020-12-28 | 2021-02-26 | 内蒙古蒙草生态环境（集团）股份有限公司 | Tissue culture method of evergreen common selfheal fruit-spike bamboo |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109452173A (en) | 2019-03-12 |
| CN109566413A (en) | 2019-04-05 |
| CN109430057A (en) | 2019-03-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103931492B (en) | The tissue culture fast seedling-cultivating method of apple rootstock M9 | |
| CN105815213A (en) | Establishing method for in-vitro regeneration system of Kiwi berry | |
| CN101946703B (en) | Method for regenerating plants of Chinese rose by using leaves as explants | |
| CN107347643A (en) | A kind of U.S.'s gold leaf Chinese honey locust tissue culture and rapid propagation method | |
| CN109757313A (en) | A kind of method for transplanting of wool savatier monochasma herb | |
| CN104542281B (en) | Mediterranean Jia Opulus tissue culture propagation method | |
| CN104304030B (en) | A kind of propagation method of oriental hybrid lily | |
| CN109392715A (en) | A kind of tissue culture and rapid propagation method of Kiwi berry gold and jade | |
| CN102823502A (en) | Method for intermediately propagating and culturing vitis quinquangularis in vitro | |
| CN106718877A (en) | A kind of flourishing torch root tuber of aromatic turmeric high quality seedling rapid propagation method | |
| CN102823503A (en) | Tissue culture medium for propagating anthurium buds by using buds | |
| CN108419675A (en) | A kind of tissue culture method of violet passion fruit top tip | |
| CN106538382B (en) | Method for establishing efficient eremochloa ophiuroides regeneration system by taking young ears as explants | |
| CN105309314A (en) | Method for setting up tissue culture regeneration system of Mongolian medicine dracocephalum moldavica | |
| CN103583360A (en) | Method for improving Abelia seedling salt tolerance by oriented induction | |
| CN106962201B (en) | A kind of pink reineckea herb test tube seedling preserving seed method | |
| CN103155868A (en) | Rapid seeding raising method of cherry rootstock ZY-1 tissue culture | |
| CN103155869A (en) | Sweet cherry rootstock Colt tissue culture method | |
| CN107683768B (en) | A kind of acclimatization and transplants method of the syringa reticulata var mandshurica tissue-cultured seedling of callus induction | |
| CN103931499B (en) | A kind of quick breeding method for tissue culture of callistemon rigidus | |
| CN1324952C (en) | Quick breeding method for tissue culture of lemon verbena | |
| CN110100740A (en) | A kind of tissue culture and rapid propagation method of two discriminations Platycerium bifurcatum | |
| CN106613973B (en) | Utilize the method for tissue-cultured seedling leaf regeneration adventitious bud fast breeding Chinese azalea | |
| CN108739403A (en) | A kind of tissue culture and rapid propagation method of rose wood | |
| CN105494105B (en) | A kind of peony tissue culture vessel seedling technology |
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 | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20171117 |