CN109845645A - A kind of culture medium of chrysanthemum method for tissue culture and tissue cultures - Google Patents
A kind of culture medium of chrysanthemum method for tissue culture and tissue cultures Download PDFInfo
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- CN109845645A CN109845645A CN201910255939.5A CN201910255939A CN109845645A CN 109845645 A CN109845645 A CN 109845645A CN 201910255939 A CN201910255939 A CN 201910255939A CN 109845645 A CN109845645 A CN 109845645A
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Abstract
The invention discloses a kind of chrysanthemum method for tissue culture and the culture mediums of tissue cultures, belong to field of plant tissue culture technique.The culture medium of tissue culture culture of the present invention includes the proliferated culture medium and root media, wherein proliferated culture medium is the MS culture medium of 0.8mg/L 6-BA+0.5mg/L NAA+0.2mg/L PVP+30g/L sucrose+6g/L agar powder, pH is 5.8, root media is the 1/2MS culture medium of 0.6mg/L NAA+0.2mg/L PVP+30g/L sucrose+6g/L agar powder, pH 5.8.Method for tissue culture of the invention has many advantages, such as that easy to operate, low in cost and chrysanthemum tissue-cultured seedling breeding rate is higher.
Description
Technical field
The invention belongs to field of plant tissue culture technique, and in particular to a kind of chrysanthemum method for tissue culture and tissue cultures
Culture medium.
Background technique
Chrysanthemum (Dendranthemamorifolium (Ramat.) Tzvel.) belongs to the perennial root grass of composite family Chrysanthemum
This plant.Chrysanthemum is one of the 10 famous flowers of China and one of the four cut-flowers in the world, is distributed across the whole world.In recent years, with
The development of world flower industry, by traditional propagation method, it is impossible to meet the market demands, and genetic transfoumation is aobvious
Shown unique superiority, can with the certain target shapes of directed modification chrysanthemum, make its pattern, the florescence, in terms of send out
Important role is waved, and successfully gene depends on the foundation of good chrysanthemum regenerating system first, it is desirable that certain regeneration
Frequency and conversion ratio, while should also have stability and repeatability.Not only breeding coefficient is high, reproduction speed is fast for tissue cultures, and
And detoxification rejuvenation, the Germ-plasma resources protection etc. of kind can be also carried out, but according to existing tissue culture technology, will appear during test
Chrysanthemum browning phenomenon, while many tissue-cultured seedling plant heights Jing Guo proliferated culture medium culture are lower, need after strong seedling culture base
It can just be inoculated into root media, extend incubation time to a certain extent, increase toxigenic capacity.
Summary of the invention
The object of the present invention is to provide one kind can effectively shorten breeding cycle, reduce breeding and cost and can effectively mention
The chrysanthemum method for tissue culture of high tissue-cultured seedling breeding rate and the culture medium of tissue cultures.
The present invention adopts the following technical scheme that achieve the above object, a kind of chrysanthemum method for tissue culture, it is characterised in that
Specific steps are as follows:
Step S1: root is gone to be divided into the high 2.5cm of stem, the stem with 1-2 piece leaf the chrysanthemum tissue-cultured seedling of healthy and strong HH-62 kind
Section, is then inoculated in proliferated culture medium and is cultivated, which is 0.8mg/L 6-BA+0.5mg/L NAA+
The MS culture medium of 0.2mg/L PVP+30g/L sucrose+6g/L agar powder, pH 5.8, condition of culture are as follows: temperature is 25 ± 2 DEG C,
Light application time 14h, intensity of illumination 2000-3000LX, cultivated days 15d, the average strain for the tissue-cultured seedling cultivated under the condition of culture
A height of 7.44cm, growth coefficient 15.36；
Step S2: choosing that growing way is consistent and the healthy and strong tissue-cultured seedling by proliferated culture medium culture, is inoculated in life after removing root
It is cultivated in root culture medium, which is 0.6mg/L NAA+0.2mg/L PVP+30g/L sucrose+6g/L agar powder
1/2MS culture medium, pH 5.8, condition of culture are as follows: temperature be 25 ± 2 DEG C, light application time 14h, intensity of illumination 2000-
3000LX, cultivated days 10d, the average root long 4.6 of the tissue-cultured seedling of the condition of culture culture, radical 12, rooting rate are
100%.
The culture medium of chrysanthemum tissue cultures of the present invention, it is characterised in that including the proliferated culture medium and culture of rootage
Base, wherein proliferated culture medium is 0.8mg/L 6-BA+0.5mg/L NAA+0.2mg/L PVP+30g/L sucrose+6g/L agar powder
MS culture medium, pH 5.8, root media is 0.6mg/L NAA+0.2mg/L PVP+30g/L sucrose+6g/L agar powder
1/2MS culture medium, pH 5.8.
The present invention mainly using MS as minimal medium, hormone 6-BA that unrooted tissue culture plant inoculation is matched in certain concentration and
, it can be achieved that shortening tissue-cultured seedling repoductive time and increasing the purpose of breeding coefficient in the proliferation and root media of NAA combination, increasing
The PVP that certain concentration is added in culture medium is grown, the generation of chrysanthemum browning phenomenon can be effectively suppressed.Chrysanthemum is during Multiplying culture
Plant height is improved, root media is directly inoculated in convenient for nursery stock, shortens repoductive time, reduce reproductive-cost.Tissue of the invention
Cultural method has many advantages, such as that easy to operate, low in cost and chrysanthemum tissue-cultured seedling breeding rate is higher.
Specific embodiment
Above content of the invention is described in further details by the following examples, but this should not be interpreted as to this
The range for inventing above-mentioned theme is only limitted to embodiment below, and all technologies realized based on above content of the present invention belong to this hair
Bright range.
Embodiment
1, the screening test of proliferated culture medium
Material to be tested is that chrysanthemum HH-62 kind removes root tissue-cultured seedling, goes root to be divided into the high 2.5cm of stem healthy and strong tissue-cultured seedling left
The right side, the stem section with 1-2 piece leaf are divided into 15 groups, every group 36, subjects are inoculated in proliferated culture medium listed by table 1
In cultivated, unified illumination cultivation condition are as follows: temperature is 25 ± 2 DEG C, light application time 14h, intensity of illumination 2000-3000LX,
Cultivated days 15d.
Proliferated culture medium is the MS of 0.2-1.0mg/L 6-BA+0.1-0.5mg/L NAA+30g/L sucrose+6g/L agar powder
Culture medium, PH 5.8.
The processing design of 1 chrysanthemum proliferated culture medium of table
2, the screening test result of proliferated culture medium
2 Multiplying culture result of table
It can be seen that the growth coefficient of processing 12 and 13 is proliferated than more prominent, but from the point of view of average plant height conducive to chrysanthemum
Culture medium should select the MS culture medium of 0.8mg/L 6-BA+0.5mg/L NAA+30g/L sucrose+6g/L agar powder, pH 5.8.
3, the brown stain test of chrysanthemum tissue-cultured seedling
Material to be tested is that chrysanthemum HH-62 kind removes root tissue-cultured seedling, is inoculated in the proliferated culture medium that table 2 filters out and is trained
It supports, while the PVP of various concentration is added, observe tissue-cultured seedling browning rate.
Unified illumination cultivation condition are as follows: temperature is 25 ± 2 DEG C, light application time 14h, intensity of illumination 2000-3000LX, culture
Number of days 15d.
The design of 3 chrysanthemum brown stain test process of table
4, chrysanthemum tissue-cultured seedling brown stain test result
4 chrysanthemum brown stain test result of table
As shown in Table 4, after 0.2mg/L PVP being added in proliferated culture medium, the production of chrysanthemum browning phenomenon can be effectively suppressed
It is raw, be conducive to optimize chrysanthemum breeding, breed high quality tissue-cultured seedling, therefore prevents the proliferated culture medium of chrysanthemum brown stain from being
The MS culture medium of 0.8mg/L 6-BA+0.5mg/L NAA+0.2mg/L PVP+30g/L sucrose+6g/L agar powder, pH 5.8.
5, the screening test of root media
Material to be tested is the tissue-cultured seedling through 4 proliferated culture medium culture of table, will choose the consistent and healthy and strong tissue culture of growing way growing way
Miao Qugen is divided into 10 groups, and subjects are inoculated in root media listed by table 5 and cultivated by every group of 36 stem sections, system
One illumination cultivation condition are as follows: temperature is 25 ± 2 DEG C, light application time 14h, intensity of illumination 2000-3000LX, cultivated days 10d.
Root media are as follows: 0.2-1.0mg/L NAA or 0.1-0.5mg/L IBA+0.2mg/L PVP+30g/L sucrose+
1/2 (N) MS culture medium of 6g/L agar powder, PH 5.8.
The test of 5 root media of table
6, the test result of different root medias
The different root media test results of table 6
As shown in Table 6, integrally reach peak-peak and be both present in processing 3, rooting rate is up to 100%, and average root is a length of
4.6cm.From addition single factors analysis, hormone NAA function and effect are better than IBA, therefore are conducive to chrysanthemum HH-62 root media and select
With 1/2 (N) MS culture medium of addition 0.6mg/L NAA+0.2mg/L PVP+30g/L sucrose+6g/L agar powder, pH 5.8.
Embodiment above describes basic principles and main features of the invention and advantage, the technical staff of the industry should
Understand, the present invention is not limited to the above embodiments, and the above embodiments and description only describe originals of the invention
Reason, under the range for not departing from the principle of the invention, various changes and improvements may be made to the invention, these changes and improvements are each fallen within
In the scope of protection of the invention.
Claims (2)
1. a kind of chrysanthemum method for tissue culture, it is characterised in that specific steps are as follows:
Step S1: going root to be divided into the high 2.5cm of stem, the stem section with 1-2 piece leaf the chrysanthemum tissue-cultured seedling of healthy and strong HH-62 kind,
Then it is inoculated in proliferated culture medium and is cultivated, which is 0.8mg/L 6-BA+0.5mg/L NAA+0.2mg/L
The MS culture medium of PVP+30g/L sucrose+6g/L agar powder, pH 5.8, condition of culture are as follows: temperature is 25 ± 2 DEG C, light application time
The average plant height of 14h, intensity of illumination 2000-3000LX, cultivated days 15d, the tissue-cultured seedling cultivated under the condition of culture is
7.44cm, growth coefficient 15.36；
Step S2: choosing that growing way is consistent and the healthy and strong tissue-cultured seedling by proliferated culture medium culture, is inoculated in training of taking root after removing root
It supports and is cultivated in base, which is the 1/ of 0.6mg/L NAA+0.2mg/L PVP+30g/L sucrose+6g/L agar powder
2MS culture medium, pH 5.8, condition of culture are as follows: temperature is 25 ± 2 DEG C, light application time 14h, intensity of illumination 2000-3000LX, training
Support number of days 10d, the average root long 4.6 of the tissue-cultured seedling of the condition of culture culture, radical 12, rooting rate 100%.
2. a kind of culture medium of chrysanthemum tissue cultures, it is characterised in that including the proliferated culture medium and root media, wherein increasing
The MS culture medium that culture medium is 0.8mg/L6-BA+0.5mg/L NAA+0.2mg/L PVP+30g/L sucrose+6g/L agar powder is grown,
PH is 5.8, and root media is that the 1/2MS of 0.6mg/L NAA+0.2mg/L PVP+30g/L sucrose+6g/L agar powder is cultivated
Base, pH 5.8.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114916440A (en) * | 2022-01-14 | 2022-08-19 | 亳州兴禾农业发展有限公司 | Tissue culture medium and tissue culture method for Bo-Chrysanthemum |
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2019
- 2019-04-01 CN CN201910255939.5A patent/CN109845645A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114916440A (en) * | 2022-01-14 | 2022-08-19 | 亳州兴禾农业发展有限公司 | Tissue culture medium and tissue culture method for Bo-Chrysanthemum |
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Application publication date: 20190607 |