CN108719072A - Rhizoma Et Radix Notopterygii seed tissue culture fast breeding technology - Google Patents
Rhizoma Et Radix Notopterygii seed tissue culture fast breeding technology Download PDFInfo
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- CN108719072A CN108719072A CN201810644125.6A CN201810644125A CN108719072A CN 108719072 A CN108719072 A CN 108719072A CN 201810644125 A CN201810644125 A CN 201810644125A CN 108719072 A CN108719072 A CN 108719072A
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- rhizoma
- radix notopterygii
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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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- 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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- 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
The invention discloses a kind of Rhizoma Et Radix Notopterygii seed tissue culture fast breeding technologies, and the Rhizoma Et Radix Notopterygii seed after Stratificated treatment is seeded in basal medium, obtains 50% or more sprout-induction rate；Then, young shoot is forwarded in proliferated culture medium, proliferating system is made to improve to 6 or so, and plant without deformity, without browning；Finally, cutting paraphyte is that single plant carries out culture of rootage.The present invention replaces the explant of existing Rhizoma Et Radix Notopterygii radical bud or spire as tissue cultures using Rhizoma Et Radix Notopterygii seed, eliminates influence of the environment temperature to explant culture, and significantly improves explant proliferation efficiency, to realize the fast breeding of Rhizoma Et Radix Notopterygii seedling.
Description
Technical field
The present invention relates to a kind of Rhizoma Et Radix Notopterygii seed tissue culture fast breeding technologies, belong to Rhizoma Et Radix Notopterygii tissue cultivating and seedling technical field.
Background technology
Rhizoma Et Radix Notopterygii is Umbelliferae Rhizoma Et Radix Notopterygii herbaceos perennial, is used as medicine with root-like stock and root, has warm-natured, bitter, Ji Nengfa
Sweat inducing diaphoresis, but can wind-damp dispelling and relieve pain, clinic be chiefly used in cold, headache and adiapneustia, wind-cold-dampness arthralgia, skeletal pain, bruise damage
The illnesss such as wound.It is mainly distributed on the alpine forest shade wetland of the extremely frigid zones of the provinces such as Sichuan, Qinghai, Gansu, Tibet, seed growing energy
Power is low, slow-growing, distribution is narrow, leads to Wild plant negligible amounts.In recent years, with the diversification of Rhizoma Et Radix Notopterygii purposes,
Its market price is constantly soaring, causes largely to excavate, keeps wild Rhizoma Et Radix Notopterygii resource endangered.
Existing Rhizoma Et Radix Notopterygii method for culturing seedlings includes mainly two kinds of forms of seed sowing and tissue cultures.But since Rhizoma Et Radix Notopterygii seed is embryo
After-ripening suspend mode type, belongs to dual suspend mode, thus under field conditions (factors), Rhizoma Et Radix Notopterygii seed need to could be sprouted through 1 year, and sprouts naturally
Hair rate is extremely low.When artificial progress seed sowing, in addition to it need to carry out Stratificated treatment breaking dormancy to seed, it must also carry out stringent and numerous
Trivial seedling management expends a large amount of manpower, causes seedling cost higher, and planting percent is extremely unstable.And tissue cultures skill
Art can be completed in a short time the cultivation of high-volume plant, thus as current Rhizoma Et Radix Notopterygii fast breeding and Germ-plasma resources protection
Important technical.If Chinese invention patent CN1918972 discloses a kind of angelica tissue culture breeding method, Chinese invention is special
Profit application CN2012101437810 discloses a kind of notopterygium forbesii tissue cultures method for in-vitro rapid propagation, Chinese invention patent
Application CN2012101437859 discloses a kind of in-vitro rapid propagation method for notopterygium tissue culture.In above-mentioned prior art
Due to, as explant, organizing to be influenced by autumn and winter low temperature or summer high temperature when Fiber differentiation using Rhizoma Et Radix Notopterygii radical bud or spire
Seriously, and its cultivation period after sprouting is long, value-added coefficient is low, to influence Rhizoma Et Radix Notopterygii tissue cultivating and seedling quality and efficiency.
Invention content
The purpose of the present invention is to provide a kind of Rhizoma Et Radix Notopterygii seed tissue culture fast breeding technologies, by the Rhizoma Et Radix Notopterygii kind after Stratificated treatment
Son is seeded in basal medium, obtains 50% or more sprout-induction rate；Then, young shoot is forwarded in proliferated culture medium, makes increasing
The system of growing is improved to 6 or so, and plant without deformity, without browning；Finally, cutting paraphyte is that single plant carries out culture of rootage.
The present invention replaces the explant of existing Rhizoma Et Radix Notopterygii radical bud or spire as tissue cultures using Rhizoma Et Radix Notopterygii seed, eliminates environment temperature pair
The influence of explant culture, and explant proliferation efficiency is significantly improved, to realize the fast breeding of Rhizoma Et Radix Notopterygii seedling.
To achieve the above object, the present invention adopts the following technical scheme that：
Rhizoma Et Radix Notopterygii seed tissue culture fast breeding technology, it is characterised in that include the following steps：
S1. the Rhizoma Et Radix Notopterygii seed progress aseptic process that level processing is completed is chosen；
S2. by step S1 treated sterile Rhizoma Et Radix Notopterygii seed is positioned on the basal medium of MS+NAA cultures to germination；
S3. by the obtained young shoots of step S2 go root go top after be forwarded on the proliferated culture medium of MS+KT+6-BA+NAA cultivate to
Form Multiple Buds；
S4. the obtained Multiple Buds of step S3 are cut into single plant, are forwarded to culture to plant on the root media of MS+NAA and give birth to
Root.
Further, the aseptic process described in step S1 includes the following steps：
S11. Rhizoma Et Radix Notopterygii seed is soaked in water to kind of skin expansion, removes the kind skin of Rhizoma Et Radix Notopterygii seed；
S12. the peeling Rhizoma Et Radix Notopterygii seed that flowing water rinsing step S11 is obtained；
S13. after use alcohol to impregnate or wipe step S12 in superclean bench treated Rhizoma Et Radix Notopterygii seed, using sterile water
Cleaning 2 ~ 3 times；
S14. a concentration of 0.1% mercuric chloride solution soaking step S13 of mass volume ratio treated Rhizoma Et Radix Notopterygii seeds are used, are then used
Sterile water wash 3 ~ 5 times.
Further, the flowing water washing time described in step S12 is no less than 30min.
Further, the ethanol postincubation time described in step S13 is 20s ~ 40s, the mercuric chloride solution described in step S14
Processing time is 5 ~ 10min.
Further, a concentration of 1.0mg/l ~ 1.5mg/l of NAA in the MS+NAA culture mediums described in step S2.
Further, a concentration of 0.5mg/l ~ 1.0mg/l of KT in the MS+KT+6-BA+NAA culture mediums described in step S3,
A concentration of 1.5mg/l ~ 2.0mg/l of 6-BA, a concentration of 0.1mg/l ~ 0.5mg/l of NAA.
Further, the young shoot height described in step S3 is 1cm ~ 1.5cm.
Further, the Multiple Buds described in step S4 remove row switching again after blade.
Further, step S2, cultivation temperature used by step S3 and step S4 is 25 ± 2 DEG C, light intensity 2000lx, light
According to time 14h/d.
Further, the culture total duration of step S3 and step S4 is no more than 60d.
Rhizoma Et Radix Notopterygii seed exosper surface horny is in flakey, uneven；Mesosperm is ossified, is unfavorable for seed suction
Rise sprouting；It greatly may be a factor for inhibiting seed to sprout that endotesta, which has,.The present invention is right before carrying out seed asepsis processing
Kind skin is soaked, is divested, and is on the one hand conducive to improve and is disinfected quality, mitigates or even sent out when avoiding later stage tissue cultures
The risk of raw infection；On the other hand sprout-induction rate after planting is made to improve to 50% or more.
The young shoot sprouted in basal medium must be to be forwarded to proliferated culture medium within the scope of 0.8cm ~ 1.5cm in height
In.If the young shoot of switching is too small, the survival rate after switching is relatively low and not easy to operate；If the young shoot of switching is excessive, mitogenetic
Proliferative capacity declines.
Basic element of cell division KT is conducive to the cell division and differentiation of Rhizoma Et Radix Notopterygii, 6-BA in the increment culture medium used in the present invention
Be conducive to bud proliferation.
Needed after Multiple Buds cutting in the present invention removal blade after again row switching, to make the small seedling rooting of single plant and leave period
Phenomenon that is synchronous, avoiding root induction rear blade withered.
Specific implementation mode
Embodiment
The specific implementation mode for elaborating the present invention in conjunction with the embodiments is as follows：
S1. the Rhizoma Et Radix Notopterygii seed that level processing is completed is chosen, is completely sunk in the water, the kind after expansion is divested after 30min
Skin.After seed kind skin is completely stripped, seed is placed under flowing water and rinses 30min.By treated, seed is arranged on filter paper,
It is transferred in superclean bench.
S2. it takes 70% alcohol to pour into beaker, seed is put into after impregnating 30s in beaker and is taken out immediately, and using sterile
Water rinses 2 ~ 3 times.After drawing excessive moisture using filter paper, then seed input is loaded in the flask of 0.1% mercuric chloride solution and is soaked
It is taken out after bubble 8min, and uses aseptic water washing 3 ~ 5 times, filter paper draws the surface of the seed moisture.
S3. configuration MS+NAA1.0mg/l culture mediums are as basic culture medium.It will complete the seed sowing of step S2 processing
To basal medium.Under 25 ± 2 DEG C of temperature, light intensity 2000lx, light application time 14h/d condition of culture, culture 30d or so, i.e.,
There can be seed sprouting successively.
S4. configuration MS+KT0.5mg/l+6-BA1.5mg/l+NAA0.1mg/l culture mediums take base as proliferated culture medium
Highly it is to be seeded on proliferated culture medium after the seed sprouting young shoot of 1cm or so goes root to go to top in basal culture medium.Temperature 25 ± 2
DEG C, under light intensity 2000lx, light application time 14h/d condition of culture, cultivate 15d ~ 25d, form more dense Multiple Buds.
S5. configuration MS+NAA0.5mg/l culture mediums are as root media.Cutting after Multiple Buds in step S4 are taken out
For single plant, it is seeded on root media respectively after cutting away blade.25 ± 2 DEG C of temperature, light intensity 2000lx, light application time 14h/
Under d condition of culture, 20d ~ 40d is cultivated, the complete root system of plant and the blade grown again can be observed.It completely regenerates to obtain the final product
Seedling.
Claims (10)
1. Rhizoma Et Radix Notopterygii seed tissue culture fast breeding technology, it is characterised in that include the following steps：
S1. the Rhizoma Et Radix Notopterygii seed progress aseptic process that level processing is completed is chosen；
S2. by step S1 treated sterile Rhizoma Et Radix Notopterygii seed is positioned on the basal medium of MS+NAA cultures to germination；
S3. by the obtained young shoots of step S2 go root go top after be forwarded on the proliferated culture medium of MS+KT+6-BA+NAA cultivate to
Form Multiple Buds；
S4. the obtained Multiple Buds of step S3 are cut into single plant, are forwarded to culture to plant on the root media of MS+NAA and give birth to
Root.
2. raising technology according to claim 1, which is characterized in that the aseptic process described in step S1 includes following step
Suddenly：
S11. Rhizoma Et Radix Notopterygii seed is soaked in water to kind of skin expansion, removes the kind skin of Rhizoma Et Radix Notopterygii seed；
S12. the peeling Rhizoma Et Radix Notopterygii seed that flowing water rinsing step S11 is obtained；
S13. after use alcohol to impregnate or wipe step S12 in superclean bench treated Rhizoma Et Radix Notopterygii seed, using sterile water
Cleaning 2 ~ 3 times；
S14. a concentration of 0.1% mercuric chloride solution soaking step S13 of mass volume ratio treated Rhizoma Et Radix Notopterygii seeds are used, are then used
Sterile water wash 3 ~ 5 times.
3. raising technology according to claim 2, which is characterized in that the flowing water washing time described in step S12 is many
In 30min.
4. raising technology according to claim 2, which is characterized in that the ethanol postincubation time described in step S13 is 20s
~ 40s, the mercuric chloride solution processing time described in step S14 are 5 ~ 10min.
5. raising technology according to claim 1, which is characterized in that NAA in the MS+NAA culture mediums described in step S2
A concentration of 1.0mg/l ~ 1.5mg/l.
6. raising technology according to claim 1, which is characterized in that the MS+KT+6-BA+NAA cultures described in step S3
A concentration of 0.5mg/l ~ 1.0mg/l of KT, a concentration of 1.5mg/l ~ 2.0mg/l of 6-BA, a concentration of 0.1mg/l ~ 0.5mg/ of NAA in base
l。
7. raising technology according to claim 1, which is characterized in that young shoot height described in step S3 be 1cm ~
1.5cm。
8. raising technology according to claim 1, which is characterized in that the Multiple Buds described in step S4 remove after blade again
Row switching.
9. raising technology according to claim 1, which is characterized in that step S2, training used by step S3 and step S4
It is 25 ± 2 DEG C, light intensity 2000lx, light application time 14h/d to support temperature.
10. raising technology according to claim 1, which is characterized in that the culture total duration of step S3 and step S4 does not surpass
Cross 60d.
Priority Applications (1)
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|---|---|---|---|
| CN201810644125.6A CN108719072A (en) | 2018-06-21 | 2018-06-21 | Rhizoma Et Radix Notopterygii seed tissue culture fast breeding technology |
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| CN201810644125.6A CN108719072A (en) | 2018-06-21 | 2018-06-21 | Rhizoma Et Radix Notopterygii seed tissue culture fast breeding technology |
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| CN201810644125.6A Pending CN108719072A (en) | 2018-06-21 | 2018-06-21 | Rhizoma Et Radix Notopterygii seed tissue culture fast breeding technology |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114176008A (en) * | 2021-12-29 | 2022-03-15 | 甘孜藏族自治州农业科学研究所 | Tissue culture breeding method for notopterygium incisum seeds |
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Address after: 641000 Hongpai Road 668, Dongxing District, Neijiang City, Sichuan Province Applicant after: Sichuan Qiancao Biotechnology Co.,Ltd. Address before: 610023 red card Road 668, Dongxing District, Neijiang, Sichuan Applicant before: SICHUAN QIANCAO BIOTECHNOLOGY Co.,Ltd. |
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Application publication date: 20181102 |