CN105830926B - A kind of quick breeding method for tissue culture of witchweed - Google Patents
A kind of quick breeding method for tissue culture of witchweed Download PDFInfo
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- CN105830926B CN105830926B CN201610232290.1A CN201610232290A CN105830926B CN 105830926 B CN105830926 B CN 105830926B CN 201610232290 A CN201610232290 A CN 201610232290A CN 105830926 B CN105830926 B CN 105830926B
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- bud
- witchweed
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- plant
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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
Abstract
The invention provides a kind of quick breeding method for tissue culture of witchweed, mainly it is used as explant by winning the axillary bud stem section of wild witchweed, then the explant Jing Guo sterilization treatment is placed in culture on inducing culture and extremely produces adventitious bud, adventitious bud is put on new inducing culture squamous subculture with evoking adventive bud fast breeding again, finally individual plant plant is put on root media and cultivated, evoking adventive bud, which is taken root, obtains amount reproduction plant.The quick breeding method for tissue culture of witchweed disclosed in this invention can make 1 axillary bud produce 10 20 sproutings in 2 months, improve the fertility of witchweed, shorten seedling raise period, save planting cost, also provide technical guarantee for the industrial seedling rearing of witchweed.
Description
Technical field
The invention belongs to field of plant tissue culture, adventitious bud is produced particular by the terminal bud or axillary bud of induction witchweed
Make witchweed rapid propagation method.
Background technology
Witchweed belongs to Scrophulariaceae unipods metal, is a kind of conventional medicinal plant, disappears infantile malnutrition due to digestive disturbances or intestinalparasites in treatment paediatrics removing heat from the liver
Aspect has very big medical value, certain to also having in terms of high liver fire, hepatitis, baby diarrhea, yctalopia, apositia, anti-inflammatory
Medical value.Witchweed is suitable to be grown in the deserted mountain meadow of low altitude area, Tian Bian, cheuch, arable land etc., and it is mainly distributed on me
Each provinces and regions in the state west and south to the southeast, but the wild witchweed resource scarcity in each area, it is impossible to meet medicinal need at this stage
Ask, this mainly due to witchweed seed it is small, germination percentage is low, slow-growing, and witchweed belongs to partial parasite, to life
Long environmental requirement is strict so that wild witchweed resource is very rare.At present, the medicinal valency of witchweed how is further developed
Value, has caused the great attention of medical field.But, what we primarily solved is the yield for how lifting witchweed, sets up a set of
The tissue culture rapid propagation technique of witchweed seedling, it is that we are badly in need of asking for solution to promote the plantation of witchweed commercial scale
Topic.
The content of the invention
In order to solve the above technical problems, the purpose of the present invention is to set up a kind of seedling tissue-culturing quick-propagation of witchweed
Method, to improve the breeding coefficient of witchweed, makes the need for seeling industry meets medicinal production.
Realize that the technical scheme that the purpose of the present invention is used is as follows：
A kind of quick breeding method for tissue culture of witchweed, it is characterised in that handled by following steps：
1) witchweed plant is plucked, segment makes every section containing an axillary-bud or top-bud；
2) by step 1) in the segment plant containing axillary-bud or top-bud be put into 0.1% mercuric chloride solution and sterilize, then with sterile
Water is cleaned three times, and 2-3min is cleaned every time；
3) by step 2 on superclean bench) in the segment plant containing axillary-bud or top-bud be inoculated into inducing culture
Cultivated, intensity is used in incubation for 1900-2100Lx artificial lighting, while control cultivation temperature to be 25-35 DEG C,
To adventitious bud is grown, then by adventitious bud be forwarded to in the new inducing culture of Fiber differentiation based component identical carry out repeatedly after
It is commissioned to train foster, the height to adventitious bud grows to 0.8-1.5cm；
4) by step 3) in adventitious bud individual plant be inoculated on root media, cultivate 3-4 weeks, plant to be planted length to 2.5-
3.5cm, its artificial lighting intensity and cultivation temperature and step 3) it is identical；
5) by step 4) in plant hardening treatment transplant for a period of time, then to artificial greenhouse, wherein artificial greenhouse
Soil is the soil mixed by peat soil and garden mould.
Preferably, the step 2) in the segment plant containing axillary-bud or top-bud be put into sterilization in 0.1% mercuric chloride solution
Time is 8-10min.
Preferably, the step 3) in inducing culture comprise the following ingredients：It is added with 1L MS culture mediums
3mg 6-BA and 0.1mg IBA, the pH that inducing culture is then adjusted with NaOH is 5.5-6.2.
Preferably, the step 4) in root media comprise the following ingredients：It is added with 1L MS culture mediums
0.1mg NAA and 0.5mg IBA, the pH that inducing culture is then adjusted with NaOH is 5.5-6.2.
Preferably, the step 5) in plant hardening treatment number of days be 5-7 days.
Preferably, the step 5) in peat soil and garden mould mass ratio be (2-4):1.
The method have the benefit that：
Stem section of the invention by cultivating the wild witchweed plant containing axillary bud on specific culture medium, establishes one
Set makes unipods payment organization rapid propagation method, improves the fertility of witchweed, shortens seedling raise period, saves planting cost,
Also technical guarantee is provided for the industrial seedling rearing of witchweed.The quick breeding method for tissue culture can make 1 axillary bud at 2 months
10-20 sprouting of interior generation, greatly improves witchweed sapling multiplication ability, for industrialization nursery and witchweed industrialized development
Provide sound assurance.And this method can whole year production seedling, contribute to witchweed plant large-scale planting, be also business
Industryization plantation provides safeguard.Meanwhile, the seed resource of wild witchweed is collected and preserved, contributes to people further to grind
Study carefully the potential medical value of wild witchweed.
Brief description of the drawings
Fig. 1 is culture figure of the axillary bud explant in inducing culture in the embodiment of the present invention one.
Fig. 2 is vegetative map of the adventitious bud in inducing culture in the embodiment of the present invention one.
Fig. 3 is culture figure of the adventitious bud in root media in the embodiment of the present invention one.
Fig. 4 is culture figure of the terminal bud explant in inducing culture in the embodiment of the present invention two.
Fig. 5 is vegetative map of the adventitious bud in inducing culture in the embodiment of the present invention two.
Fig. 6 is culture figure of the adventitious bud of the embodiment of the present invention two in root media.
Fig. 7 is the hardening state diagram of plant in the embodiment of the present invention two.
Embodiment
Below, with reference to accompanying drawing and embodiment, the present invention is described further：
Embodiment one
A kind of quick breeding method for tissue culture of witchweed, it is characterised in that handled by following steps：
1) wild witchweed plant is plucked, segment makes every section containing an axillary bud；
2) by step 1) in the segment plant containing axillary bud be put into 0.1% mercuric chloride solution and sterilize 8-10min, then with sterile
Water is cleaned three times, and 2-3min is cleaned every time；
3) by step 2 on superclean bench) in the segment plant containing axillary bud be inoculated into inducing culture and trained
Support, wherein inducing culture is comprised the following ingredients：3mg 6-BA and 0.1mg IBA are added with 1L MS culture mediums, then
The pH that inducing culture is adjusted with NaOH is 5.5-6.2, is 1900-2100Lx in artificial lighting intensity, temperature is at 25-35 DEG C
Cultivated, until growing adventitious bud, be then forwarded to adventitious bud and the new inducing culture of Fiber differentiation based component identical
It is middle to carry out multiple squamous subculture, highly grow to 0.8-1.5cm to adventitious bud；
4) by step 3) in adventitious bud individual plant be inoculated on root media, wherein root media is by following component
Composition：0.1mg NAA and 0.5mg IBA are added with 1L MS culture mediums, are then with the NaOH pH for adjusting root media
5.5-6.2, cultivates 3-4 week, 2.5-3.5cm is grown to plant height, wherein, artificial lighting intensity in the incubation with
Cultivation temperature and step 3) it is identical；
5) by step 4) in plant hardening treatment 5-7 days, then transplant to the soil of artificial greenhouse, wherein artificial greenhouse
It is the soil mixed by peat soil and garden mould, wherein the mass ratio of peat soil and garden mould is (2-4):1.
Fig. 1-Fig. 3 is the cultivation state diagram of the present embodiment：Wherein, Fig. 1 is using the axillary bud stem section of wild witchweed as outer
Implant, it is sterilized after, explant is inoculated into the cultivation figure of inducing culture on superclean bench；As it is clear from fig. 2 that axillary bud stem
Section passes through a large amount of adventitious buds of multiple squamous subculture hyperplasia in inducing culture；Fig. 3 is adventitious bud individual plant in root media
On grow root system, form regeneration plant.
Embodiment two
A kind of quick breeding method for tissue culture of witchweed, it is characterised in that handled by following steps：
1) wild witchweed plant is plucked, segment makes every section containing a terminal bud；
2) by step 1) in the segment plant containing terminal bud be put into 0.1% mercuric chloride solution and sterilize 8-10min, then with sterile
Water is cleaned three times, and 2-3min is cleaned every time；
3) by step 2 on superclean bench) in the segment plant containing terminal bud be inoculated into inducing culture and trained
Support, wherein inducing culture is comprised the following ingredients：3mg 6-BA and 0.1mg IBA are added with 1L MS culture mediums, then
The pH that inducing culture is adjusted with NaOH is 5.5-6.2, is 1900-2100Lx in artificial lighting intensity, temperature is at 25-35 DEG C
Cultivated, until growing adventitious bud, be then forwarded to adventitious bud and the new inducing culture of Fiber differentiation based component identical
It is middle to carry out multiple Fiber differentiation, highly grow to 0.8-1.5cm to adventitious bud；
4) by step 3) in adventitious bud individual plant be inoculated on root media, wherein root media is by following component
Composition：0.1mg NAA and 0.5mg IBA are added with 1L MS culture mediums, are then with the NaOH pH for adjusting inducing culture
5.5-6.2, is cultivated 3-4 week, plant to be planted length to 2.5-3.5cm, wherein, artificial lighting intensity in the incubation and culture are warm
Degree and step 3) it is identical；
5) by step 4) in plant hardening treatment 5-7 days, then transplant to the soil of artificial greenhouse, wherein artificial greenhouse
It is the soil mixed by peat soil and garden mould, wherein the mass ratio of peat soil and garden mould is (2-4):1.
Such as the cultivation state diagram that Fig. 4-7 is the present embodiment：Fig. 4 be the terminal bud stem section using wild witchweed as explant,
After sterilized, explant is inoculated into the cultivation figure of inducing culture on superclean bench；Fig. 5 understands that explant is in induction
Pass through a large amount of adventitious buds of multiple squamous subculture hyperplasia in culture medium；Fig. 6 is that individual plant adventitious bud is grown on root media
Root system, forms regeneration plant；Fig. 7 is the process that hardening treatment is carried out to regeneration plant, plant is adapted to rapidly after being colonized
Unsuitable environmental condition, shortens transplanting seedling time, strengthens the resistivity of plant pair low temperature, strong wind etc..
For those skilled in the art, technical scheme that can be as described above and design, make other each
It is kind corresponding to change and deform, and all these change and deformation should all belong to the protection model of the claims in the present invention
Within enclosing.
Claims (4)
1. a kind of quick breeding method for tissue culture of witchweed, it is characterised in that handled by following steps：
1）Witchweed plant is plucked, segment makes every section containing an axillary-bud or top-bud；
2）By step 1）In the segment plant containing axillary-bud or top-bud be put into 0.1% mercuric chloride solution and sterilize, then use sterile water wash
Three times, 2-3min is cleaned every time；
3）By step 2 on superclean bench）In the segment plant containing axillary-bud or top-bud be inoculated into inducing culture carry out
Intensity is used for 1900-2100Lx artificial lighting in culture, incubation, while controlling cultivation temperature to be 25-35 DEG C, to length
Go out adventitious bud, then adventitious bud is forwarded to carrying out multiple subculture training in the new inducing culture of Fiber differentiation based component identical
Support, the height to adventitious bud grows to 0.8-1.5cm；
The step 3）In inducing culture comprise the following ingredients：In 1L MS culture mediums added with 3mg 6-BA and
0.1mg IBA, the pH that inducing culture is then adjusted with NaOH is 5.5-6.2；
4）By step 3）In adventitious bud individual plant be inoculated on root media, cultivate 3-4 weeks, plant to be planted length to 2.5-3.5cm,
Its artificial lighting intensity and cultivation temperature and step 3）It is identical；
The step 4）In root media comprise the following ingredients：In 1L MS culture mediums added with 0.1mg NAA and
0.5mg IBA, the pH that inducing culture is then adjusted with NaOH is 5.5-6.2；
5）By step 4）In plant hardening treatment transplant for a period of time, then to the soil of artificial greenhouse, wherein artificial greenhouse
It is the soil mixed by peat soil and garden mould.
2. the quick breeding method for tissue culture of witchweed according to claim 1, it is characterised in that the step 2）In
The disinfecting time that segment plant containing axillary-bud or top-bud is put into 0.1% mercuric chloride solution is 8-10min.
3. the quick breeding method for tissue culture of witchweed according to claim 1, it is characterised in that the step 5）In
Plant hardening treatment number of days is 5-7 days.
4. the quick breeding method for tissue culture of witchweed according to claim 1, it is characterised in that the step 5）In
The mass ratio of peat soil and garden mould is（2-4）:1.
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| CN201610232290.1A CN105830926B (en) | 2016-04-13 | 2016-04-13 | A kind of quick breeding method for tissue culture of witchweed |
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| CN105830926B true CN105830926B (en) | 2017-10-24 |
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| CN112425506A (en) * | 2020-11-23 | 2021-03-02 | 广东省农业科学院作物研究所 | Tissue culture rapid propagation method of artemisia anomala |
| CN115868410B (en) * | 2022-12-06 | 2024-02-09 | 赣南医学院 | Tissue culture method for promoting rapid germination of striga oleracea seeds |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102144567A (en) * | 2011-04-21 | 2011-08-10 | 肇庆学院 | Rapid propagation method of torenia flava |
| CN104365483A (en) * | 2014-11-20 | 2015-02-25 | 安徽省农业科学院园艺研究所 | Scrophularia ningpoensis hemsl breeding method |
| CN104663446A (en) * | 2015-02-25 | 2015-06-03 | 杨惠才 | In-vitro rapid propagation method of penstemon barbatus |
| CN105309311A (en) * | 2015-11-06 | 2016-02-10 | 道真仡佬族苗族自治县万福农业综合开发有限公司 | Method for breeding improved variety of scrophularia ningpoensis Hemsl. |
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2016
- 2016-04-13 CN CN201610232290.1A patent/CN105830926B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102144567A (en) * | 2011-04-21 | 2011-08-10 | 肇庆学院 | Rapid propagation method of torenia flava |
| CN104365483A (en) * | 2014-11-20 | 2015-02-25 | 安徽省农业科学院园艺研究所 | Scrophularia ningpoensis hemsl breeding method |
| CN104663446A (en) * | 2015-02-25 | 2015-06-03 | 杨惠才 | In-vitro rapid propagation method of penstemon barbatus |
| CN105309311A (en) * | 2015-11-06 | 2016-02-10 | 道真仡佬族苗族自治县万福农业综合开发有限公司 | Method for breeding improved variety of scrophularia ningpoensis Hemsl. |
Non-Patent Citations (1)
| Title |
|---|
| 独脚金的研究概述;龚明等;《药物研究》;20101231(第4期);第930-931页 * |
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