CN106171976A - A kind of Ramulus Sambuci Williamsii tissue culture and rapid propagation method - Google Patents
A kind of Ramulus Sambuci Williamsii tissue culture and rapid propagation method Download PDFInfo
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- CN106171976A CN106171976A CN201610530411.0A CN201610530411A CN106171976A CN 106171976 A CN106171976 A CN 106171976A CN 201610530411 A CN201610530411 A CN 201610530411A CN 106171976 A CN106171976 A CN 106171976A
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- culture
- ramulus sambuci
- sambuci williamsii
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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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- Developmental Biology & Embryology (AREA)
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Abstract
The present invention relates to field of plant growing technology, be specifically related to a kind of Ramulus Sambuci Williamsii tissue culture and rapid propagation method.The present invention uses the method for tissue culture, obtains, by outer implant of sterilizing, inducing culture, enrichment culture, root culture, the plant that takes root, field planting land for growing field crops after seedling exercising is tamed.Innovative point is that the Ramulus Sambuci Williamsii tissue culture and rapid propagation method of the present invention can breed Ramulus Sambuci Williamsii tissue cultured seedling with rapid, high volume, to adapt to the needs of afforestation, woody edible oil.
Description
Technical field:
The present invention relates to field of plant growing technology, be specifically related to a kind of Ramulus Sambuci Williamsii tissue culture and rapid propagation method.
Background technology:
In recent years, the demand sustainable growth of China's edible vegetable oil, demand gap constantly expands, and external dependence degree is obvious
Rising, security issues become increasingly urgent for edible vegetable oil.Woody oleiferous plants industry is the conventional industries of China, is also to provide healthy high-quality
The important sources of edible oil, and Ramulus Sambuci Williamsii (Sambucua williamsii Hance) is the woody of a kind of great Development volue
One of oil plant seeds, research confirms that its fruit oil content reaches 35%～44%, and the saturated fatty acid content contained is low, human body must
Health, both relative to soybean oil, Oleum Arachidis hypogaeae semen, Helianthi innage, is highly profitable by the content of fatty acid of palpus.
Ramulus Sambuci Williamsii modes of reproduction is frequently with seminal propagation, it is also possible to cuttage and propagation by grafiting, but, utilize traditional approach numerous
Growing, it is relatively big by being affected season, breeds speed relatively slowly, and therefore research Ramulus Sambuci Williamsii rapid propagation in vitro method, improves synthetism further
The exploitation of wood, value, to Ramulus Sambuci Williamsii large-scale production and the Rapid Popularization of new quality product kind, have important theory and reality
Meaning.
Summary of the invention
It is an object of the invention to provide the technology of a kind of Ramulus Sambuci Williamsii tissue-culturing rapid propagation, it includes following step:
1): stem segment with axillary bud is disinfected
Selecting Ramulus Sambuci Williamsii to give birth to the tender point of the full branch of internal organs bud then, be cut into 4-5cm segment, every section contains 1-2 axillalry bud, stream
Rinse 30min～60min, with 75% alcohol-pickled 30s on superclean bench, aseptic water washing 2 times under water, then use respectively
0.1%HgCl2Processing 15min, sterilized water vibration is standby after rinsing 3～5 times；
2): induced bundle is sprouted
By step 1) the sterilization wound at stem segment with axillary bud two ends after the sterilization of gained cuts away, and stem section controls 2～3cm,
Guarantee every section of axillalry bud 1 having for inducing startup, access on the MS inducing culture of volume 30-50ml, every bottle graft kind 1-2
The individual outer implant containing axillalry bud, first light culture 3～5 days under the conditions of 23～27 DEG C, be subsequently placed in illumination every day 10～12 hours, light
It is that light is cultivated under the conditions of 1500～2000lx according to intensity, until induced synthesis Multiple Buds；
3): differentiation culture
By step 2) Multiple Buds that induced proceeds to the propagation being made up of MS+6BA1mg/L+TDZ0.01mg/L of 50ml
In culture medium, 5 sprouts of every bottle of switching carry out enrichment culture, until growth coefficient is 4, every strain pumping goes out 2-3 sections, plant height
Reach 3-5cm, obtain differentiation Seedling；
4): root culture
By step 3) the differentiation Seedling of gained turns that to be inoculated into volume be the root media that 50ml is made up of MS+IBA0.1mg/L
On carry out root culture, every bottle of switching 5 strains, root plant height to be generated to 5-7cm, root system more than 5, healthy and strong vibrant, Ji Kejin
Row rooting culture；
5): seedling exercising is tamed
By step 4) plant that takes root that obtains, seedling exercising 7-10 days, moving to equipped with perlite and turfy soil volume ratio is 3:5's
It is mixed in the nutrient cup of substrate, Field planting after continuing to cultivate 25 days.
Preferably, step 1) in, Ramulus Sambuci Williamsii gives birth to the tender point of the full branch of internal organs bud then, is cut into 5cm segment, and every section contains 1
Individual axillalry bud.
Preferably, step 2) described in illumination every day 10 hours, intensity of illumination is 2000lx.
The present invention uses the method for tissue culture, is obtained by outer implant of sterilizing, inducing culture, enrichment culture, root culture
To the plant that takes root, field planting land for growing field crops after seedling exercising is tamed.Innovative point be the present invention Ramulus Sambuci Williamsii tissue culture and rapid propagation method can be fast
Speed amount reproduction Ramulus Sambuci Williamsii tissue cultured seedling, to adapt to the needs of afforestation, woody edible oil, and it is similar to have no that other team have
Report.
Detailed description of the invention
Embodiment 1
Described Ramulus Sambuci Williamsii group culturation rapid propagating technology, specifically comprises the following steps that
First, select Ramulus Sambuci Williamsii to give birth to the tender point of the full branch of internal organs bud then, gather outer implant.The outer implant that will gather, cuts
Becoming 4.5cm segment, every section contains 1 axillalry bud, rinses after 45min under flowing water, with 75% alcohol-pickled on superclean bench
30s, aseptic water washing 2 times, then use 0.1%HgCl respectively2Processing 15min, sterilized water vibration is rinsed 5 times；
Second: will disinfect the stem segment with axillary bud obtained and cut away the sterilization wound at stem section two ends, stem section controls
2.5cm, accesses in MS culture medium, and first full light culture 5 days under the conditions of 23 DEG C, are subsequently placed in illumination every day 12 hours, and illumination is strong
Degree is 1500lx, until induced synthesis Multiple Buds；
3rd: the Multiple Buds induced is proceeded to the propagation being made up of the MS+6-BA1mg/L+TDZ0.01mg/L training of 50ml
Support and carry out enrichment culture on base；
4th: be inoculated on root media by differentiation Seedling and carry out root culture, root media is MS+IBA0.1mg/
L；
5th: after the bottle Seedling seedling exercising 10 days of reincarnation root, plant into being the mixing of 3:5 equipped with perlite and turfy soil volume ratio
Become in the nutrient cup of substrate, obtain seedling, Field planting after cultivating 25 days.
This embodiment during carrying out group training to Ramulus Sambuci Williamsii, and the success rate of outer implant sterilization is 95% outer implant induced bundle
Production rate of sprouting is 96%, and the growth coefficient of Multiple Buds is 4, breaks up Seedling root induction rate 100%, and the bottle Seedling seedling exercising of reincarnation root is tamed and dociled
Change survival rate 98%.
The Ramulus Sambuci Williamsii tissue cultured seedling field planting land for growing field crops obtained in embodiment 1 after 2 months to its increment with as 2, field planting land for growing field crops
The tree seedling of the moon is contrasted, and contrasts table such as table 1 below
Difference condition table behind table 1, Ramulus Sambuci Williamsii tissue cultured seedling and tree seedling field planting land for growing field crops
Embodiment 2
Described Ramulus Sambuci Williamsii group culturation rapid propagating technology, specifically comprises the following steps that
First, select Ramulus Sambuci Williamsii to give birth to the tender point of the full branch of internal organs bud then, gather outer implant.The outer implant that will gather, cuts
Becoming 4cm segment, every section contains 1 axillalry bud, after rinsing 40min under flowing water, with 75% alcohol-pickled 30s on superclean bench,
Aseptic water washing 2 times, then use 0.1%HgCl respectively2Processing 15min, sterilized water vibration is rinsed 4 times；
Second: will disinfect the stem segment with axillary bud obtained and cut away the sterilization wound at stem section two ends, stem section controls at 3cm,
Accessing in MS culture medium, first full light culture 5 days under the conditions of 25 DEG C, are subsequently placed in illumination every day 11 hours, and intensity of illumination is
1800lx, until induced synthesis Multiple Buds；
3rd: the Multiple Buds induced is proceeded to the propagation being made up of the MS+6-BA1mg/L+TDZ0.01mg/L training of 50ml
Support and carry out enrichment culture on base；
4th: be inoculated on root media by differentiation Seedling and carry out root culture, root media is MS+IBA0.1mg/
L；
5th: after the bottle Seedling seedling exercising 8 days of reincarnation root, plant into being the mixing of 3:5 equipped with perlite and turfy soil volume ratio
Become in the nutrient cup of substrate, obtain seedling, Field planting after cultivating 25 days.
This embodiment during carrying out group training to Ramulus Sambuci Williamsii, and the success rate of outer implant sterilization is 92%, and outer implant is induced
Multiple Buds production rate is 96%, and the growth coefficient of Multiple Buds is 4, breaks up Seedling root induction rate 100%, the bottle Seedling seedling exercising of reincarnation root
Domestication survival rate 97%.
The Ramulus Sambuci Williamsii tissue cultured seedling field planting land for growing field crops obtained in embodiment 2 after 2 months to its increment with as 4, field planting land for growing field crops
The tree seedling of the moon is contrasted, and contrasts table such as table 2 below
Difference condition table behind table 2, Ramulus Sambuci Williamsii tissue cultured seedling and tree seedling field planting land for growing field crops
Embodiment 3
Described Ramulus Sambuci Williamsii group culturation rapid propagating technology, specifically comprises the following steps that
First, select Ramulus Sambuci Williamsii to give birth to the tender point of the full branch of internal organs bud then, gather outer implant.The outer implant that will gather, cuts
Becoming 5cm segment, every section contains 1 axillalry bud, after rinsing 50min under flowing water, with 75% alcohol-pickled 30s on superclean bench,
Aseptic water washing 2 times, then use 0.1%HgCl respectively2Processing 15min, sterilized water vibration is rinsed 4 times；
Second: will disinfect the stem segment with axillary bud obtained and cut away the sterilization wound at stem section two ends, stem section controls, at 3m, to connect
Entering in MS culture medium, first full light culture 5 days under the conditions of 27 DEG C, are subsequently placed in illumination every day 10 hours, and intensity of illumination is
2000lx, until induced synthesis Multiple Buds；
3rd: the Multiple Buds induced is proceeded to the propagation being made up of the MS+6-BA1mg/L+TDZ0.01mg/L training of 50ml
Support and carry out enrichment culture on base；
4th: be inoculated on root media by differentiation Seedling and carry out root culture, root media is MS+IBA0.1mg/
L；
5th: after the bottle Seedling seedling exercising 9 days of reincarnation root, plant into being the mixing of 3:5 equipped with perlite and turfy soil volume ratio
Become in the nutrient cup of substrate, obtain seedling, Field planting after cultivating 25 days.
This embodiment during carrying out group training to Ramulus Sambuci Williamsii, and the success rate of outer implant sterilization is 92%, and outer implant is induced
Multiple Buds production rate is 96%, and the growth coefficient of Multiple Buds is 4, breaks up Seedling root induction rate 100%, the bottle Seedling seedling exercising of reincarnation root
Domestication survival rate 97%.
The Ramulus Sambuci Williamsii tissue cultured seedling field planting land for growing field crops obtained in embodiment 3 after 2 months to its increment with as 4, field planting land for growing field crops
The tree seedling of the moon is contrasted, and contrasts table such as table 3 below
Difference condition table behind table 3, Ramulus Sambuci Williamsii tissue cultured seedling and tree seedling field planting land for growing field crops
Conclusion: using after tissue-culturing rapid propagation after technology, the success rate of outer implant sterilization, outer implant induced bundle are sprouted generation
Rate, differentiation Seedling root induction rate, reincarnation root bottle Seedling seedling exercising domestication survival rate the highest, the growth coefficient of Multiple Buds reaches 4；Group
Behind the seedling field planting land for growing field crops that training is cultivated, its plant height, hat width, the increment in footpath be all higher than tree seedling increment.
Experimental result: use Ramulus Sambuci Williamsii quick-breeding method of the present invention, can be effectively improved outer implant sterilization success rate, outer
Implant induced bundle sprout production rate, differentiation Seedling root induction rate, reincarnation root bottle Seedling seedling exercising domestication survival rate.
Claims (3)
1. a Ramulus Sambuci Williamsii tissue culture and rapid propagation method, it is characterised in that specifically comprise the following steps that
1): stem segment with axillary bud is disinfected
Selecting Ramulus Sambuci Williamsii to give birth to the tender point of the full branch of internal organs bud then, be cut into 4-5cm segment, every section contains 1-2 axillalry bud, under flowing water
Rinse 30min～60min, with 75% alcohol-pickled 30s on superclean bench, aseptic water washing 2 times, more respectively with 0.1%
HgCl2After processing 15min, aseptic water washing 3～5 times standby；
2): induced bundle is sprouted
By step 1) the sterilization wound at stem segment with axillary bud two ends after the sterilization of gained cuts away, and stem section controls 2～3cm, it is ensured that
Every section has the axillalry bud 1 for inducing startup, accesses on the MS inducing culture of volume 30-50ml, and every bottle graft kind 1-2 is containing axil
The outer implant of bud, first full light culture 3～5 days under the conditions of 23～27 DEG C, be subsequently placed in illumination every day 10～12 hours, and illumination is strong
Degree is light cultivation under the conditions of 1500～2000lx, until induced synthesis Multiple Buds；
3): differentiation culture
By step 2) Multiple Buds that induced proceeds to the enrichment culture being made up of MS+6BA1mg/L+TDZ0.01mg/L of 50ml
On base, 5 sprouts of every bottle of switching carry out enrichment culture, until growth coefficient is 4, every strain pumping goes out 2-3 sections, and plant height reaches 3-
5cm, obtains differentiation Seedling；
4): root culture
By step 3) the differentiation Seedling of gained turns that to be inoculated into volume be that the root media that 50ml is made up of MS+IBA0.1mg/L is enterprising
Row root culture, every bottle of switching 5 strains, root plant height to be generated to 5-7cm, root system more than 5, healthy and strong vibrant, can tame and docile
Change and transplant；
5): seedling exercising is tamed
By step 4) plant that takes root that obtains, seedling exercising 7-10 days, moving to equipped with perlite and turfy soil volume ratio is the mixing of 3:5
Become in the nutrient cup of substrate, Field planting after continuing to cultivate 25 days.
2. Ramulus Sambuci Williamsii tissue culture and rapid propagation method as claimed in claim 1, it is characterised in that step 1) in, described Ramulus Sambuci Williamsii is worked as
The tender point of the year life full branch of internal organs bud, is cut into 5cm segment, and every section contains 1 axillalry bud.
3. Ramulus Sambuci Williamsii tissue culture and rapid propagation method as claimed in claim 1, it is characterised in that step 2) described in illumination every day 10
Hour, intensity of illumination is 2000lx.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610530411.0A CN106171976A (en) | 2016-07-07 | 2016-07-07 | A kind of Ramulus Sambuci Williamsii tissue culture and rapid propagation method |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610530411.0A CN106171976A (en) | 2016-07-07 | 2016-07-07 | A kind of Ramulus Sambuci Williamsii tissue culture and rapid propagation method |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112715361A (en) * | 2020-12-30 | 2021-04-30 | 美尚生态景观股份有限公司 | Culture method of black pagoda elderberry subculture seedlings |
| CN112715362A (en) * | 2020-12-31 | 2021-04-30 | 黑龙江省林业科学研究所 | Tissue culture method of wild northeast elderberry |
| CN112753574A (en) * | 2020-12-30 | 2021-05-07 | 美尚生态景观股份有限公司 | Method for rooting in black pagoda elderberry tissue culture seedling bottle |
-
2016
- 2016-07-07 CN CN201610530411.0A patent/CN106171976A/en active Pending
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112715361A (en) * | 2020-12-30 | 2021-04-30 | 美尚生态景观股份有限公司 | Culture method of black pagoda elderberry subculture seedlings |
| CN112753574A (en) * | 2020-12-30 | 2021-05-07 | 美尚生态景观股份有限公司 | Method for rooting in black pagoda elderberry tissue culture seedling bottle |
| CN112715362A (en) * | 2020-12-31 | 2021-04-30 | 黑龙江省林业科学研究所 | Tissue culture method of wild northeast elderberry |
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Application publication date: 20161207 |
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