CN105724255A - Tissue culture medium capable of improving davidia involucrata differentiation and germination - Google Patents
Tissue culture medium capable of improving davidia involucrata differentiation and germination Download PDFInfo
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- CN105724255A CN105724255A CN201610268650.3A CN201610268650A CN105724255A CN 105724255 A CN105724255 A CN 105724255A CN 201610268650 A CN201610268650 A CN 201610268650A CN 105724255 A CN105724255 A CN 105724255A
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- culture medium
- tissue culture
- tcm
- dove tree
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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
Abstract
The invention belongs to the technical field of tissue culture production, and particularly relates to a tissue culture medium capable of improving davidia involucrata differentiation and germination.The tissue culture medium is characterized by being prepared from, by weight, 4.85-8.65 g/L of MS, 0.3-1.5 mg/L of NAA, 0.3-1.2 mg/L of IBA, 0.5-1.8 g/L of activated carbon, 0.15-0.20 mg/L of metalaxyl, 0.3-0.8 mg/L of indole-3-butyric acid potassium salt, 0.2-0.4 mg/L of forchlorfenuron, 0.04-0.10 mg/L of microcystin, 0.15-0.25 mg/L of diethyl aminoethyl hexanoate, 0.08-0.15 mg/L of putrescine, 25-30 g/L of saccharose and 8-15 g/L of agar.By means of the tissue culture medium capable of improving davidia involucrata differentiation and germination, a good growing environment can be provided for callus of davidia involucrata, the source of raw materials is renewable, the price is low, and industrial production is promoted.
Description
[technical field]
The invention belongs to tissue culture's production technical field, be specifically related to a kind of improve dove tree and break up the tissue culture medium (TCM) that sprouts.
[background technology]
Dove tree is deciduous tree.Can grow into 15～25 meters high, leaf is the most avette, and there is sawtooth at edge.Graminaceous plant only has a genus
Two kinds, two kinds similar, a kind of blade face hairiness, and another kind of davidia involucrata var. vilmoriniana is bright finish.Color spends strange U.S., is new before 10,000,000 years
Raw for the Relict Plant stayed the Tertiary Period, in quaternary glacier period, the dove tree of most area becomes extinct in succession, only in China
More southern areas survive, Luoyang green sincere agricultural propagation in scale and planting successfully.Become " living of plant kingdom's today
Fossil ", it is described as " the pigeon tree of China ", also known as " pigeon flower tree ", " water pear ", wild species are only grown in China
Southwest Sichuan Province and middle part Hubei Province and surrounding area.
Dove tree has the title of " plant living fossil ", is the treasure in 8 kinds of one-level national key protected plant of country, because its flower-shape exactly likes exhibition
Pigeon that wing circles in the air and by west botanist named " China pigeon tree ".Owing to the felling of forest destroys and excavates seeding-growth
Planting, negligible amounts, distribution reduces the most day by day, if not taking protective measure, has the danger substituted by other deciduous species
Danger.Dove tree is the deleted species away from modern 60,000,000 years Pre-Cenozoic Gu in Tertiary Period tropical plants faunas.It not only has the valency of viewing and admiring
Value: dove tree is that world-renowned preciousness views and admires tree, often plants in side, pond, small stream is taken up near sanitarium, hotel, exhibition center, and had
Symbol of peace meaning；And there is economic worth: material is heavy, is the first-class material of building, can make furniture and carve
Material.Though built nature reserve area, some areal area, but without strict protection measure, protection point also should be set in other areal area,
Concrete conservative management measure should be formulated, actively develop introducing and planting and reproduction test, carry out artificial afforestration, expand its distribution
District.
Having now and bred dove tree by graft technology, but responding time is long, test planting percent is low, so also having now
Occur that the propagation method of tissue culture processes, to improve the breeding potential of dove tree, but during group training, inducing culture usually occurs
Bud ratio relatively low, affect the growth in later stage, cause emergence rate to reduce, in order to prevent the generation of situation in this, it is necessary in group
Training early stage is ready, it is ensured that callus breaks up the bud ratio sprouted, and is the key point of the breeding improving the training of dove tree group.
[summary of the invention]
It is an object of the invention to: for the problem of above-mentioned existence, it is provided that a kind of improve dove tree and break up the tissue culture medium (TCM) that sprouts,
Can provide dove tree callus one good growing environment, and raw material sources are renewable, cheap, it is beneficial to industry metaplasia
Produce.
To achieve these goals, the technical solution used in the present invention is as follows:
A kind of improving dove tree and break up the tissue culture medium (TCM) that sprouts, the raw material of described tissue culture medium (TCM) is in parts by weight: MS
4.85-8.65g/L+NAA 0.3-1.5mg/L+IBA 0.3-1.2mg/L+ activated carbon 0.5-1.8g/L+ metalaxyl 0.15-0.20mg/L+ Yin
Diindyl potassium butyrate 0.3-0.8mg/L+ forchlorfenuron 0.2-0.4mg/L+ microcystin 0.04-0.10mg/L+ diethyl aminoethyl hexanoate 0.15-0.25mg/L+
Putrescine 0.08-0.15mg/L, sucrose 25-30g/L, agar 8-15g/L.
Preferably, described tissue culture medium (TCM) is in parts by weight: MS 5.0-7.50g/L+NAA 0.5-1.20mg/L+IBA
0.5-0.85mg/L+ activated carbon 0.6-1.30g/L+ metalaxyl 0.17-0.19mg/L+ butyric acid potassium 0.45-0.65mg/L+ forchlorfenuron
0.22-0.35mg/L+ microcystin 0.05-0.09mg/L+ diethyl aminoethyl hexanoate 0.16-0.20mg/L+ putrescine 0.010-0.13mg/L, sucrose
26-19g/L, agar 8-15g/L.
More preferably, described tissue culture medium (TCM) is in parts by weight: MS 5.25g/L+NAA 0.85mg/L+IBA
0.58mg/L+ activated carbon 0.77g/L+ metalaxyl 0.18mg/L+ butyric acid potassium 0.55mg/L+ forchlorfenuron 0.25mg/L+ microcystin
0.06mg/L+ diethyl aminoethyl hexanoate 0.18mg/L+ putrescine 0.12mg/L, sucrose 28g/L, agar 11g/L.
In the present invention, as further illustrating, the pH 5.8-6.4 of described tissue culture medium (TCM).
In the present invention, as further illustrating, the granularity of the activated carbon of described tissue culture medium (TCM) is 100-200 mesh.
In the present invention, as further illustrating, described tissue culture medium (TCM) also includes that the later stage processes, and is former by tissue culture medium (TCM)
The culture medium configured, according to parts by weight, is then put into high temperature sterilize pot, at 131 DEG C, 1.2kg/cm by material2Under the conditions of enter
Row sterilizing 35-45min, then takes out and is cooled to room temperature, puts into topple in aseptic operating platform to be dispensed into and passes through high temperature sterilize pot
In the culture dish of sterilizing.
In sum, owing to have employed technique scheme, the invention has the beneficial effects as follows:
The present invention further studies on the formula of traditional minimal medium, adds some and ensure that dove tree has one
Individual excellent growing conditions, through research find use the present invention basic culture solution after add 6-BA+NAA+IBA+ activated carbon+
Metalaxyl+butyric acid potassium+forchlorfenuron+microcystin+diethyl aminoethyl hexanoate+putrescine, it is possible to promote the division of dove tree cell, accelerate it and go out
The time of bud, and a bud gone out is disease-free, color is glossy dark green, and bud meat thickness is full, and bud ratio reaches 100%.Wherein, interpolation is lived
Property charcoal is prevented from callus and occurs that browning, auxin naphthalene acetic acid and mitogen butyric acid potassium cooperate, and promotes to plant
The differentiation of thing vascular tissue, and use forchlorfenuron further to promote that histiocyte divides, fast relative to generally using 6-benzyl amino
Purine, split speed exceeds 10-100 times, accelerates cell mitogen, promotes cell to increase and differentiation, rich in having in microcystin
Active oxygen, in plant, to mediate multiple response in plant cell signaling transduction pathway as second message,second messenger anti-for the active oxygen of low concentration
Should, but the active oxygen of high concentration then causes the oxidative damage even cell death of biomacromolecule, through experimental study at formula
Middle interpolation microcystin 0.06mg/L, it is ensured that the active oxygen of plant low concentration can be supplied to, will not damaging cells, play second
The effect of courier, promotes that auxin flows to botanical system, is provided that callus nutrition；Good life can be had in culture medium
In the case of long environment, and then in culture medium, add putrescine so that being absorbed by histiocyte of putrescine Energy velocity, adjust
PH value in ganglion cell, can be combined with the polyamines regulatory site of nmda receptor, strengthens the generation of NMDA induction, regulation cell
Propagation and differentiation function；The present invention has also compounded diethyl aminoethyl hexanoate, it is possible to will quickly break up the tissue after sprouting, it is provided that follow-up nutrition is propped up
Hold, promote that tissue improves peroxidase and the activity of nitrate reductase, improve chlorophyllous content and accelerate photosynthetic speed, promote
The division of plant cell and elongation, promote the growth of root system, regulate the balance of internal nutrient, sprouts period to prevent at tissue
Fungal infection, the further present invention has also compounded metalaxyl, can not only effectively suppress the mycelial normal life of pathogenic fungi
Grow or directly kill pathogenic bacteria, along with the absorption of water flow in seed cell and good in the stable under acidic conditions performance of the application,
Ensure that its performance time is long, prevent aetiolation, improve the differentiation bud ratio of dove tree.
[detailed description of the invention]
Embodiment 1:
A kind of improving dove tree and break up the tissue culture medium (TCM) that sprouts, the raw material of described tissue culture medium (TCM) is in parts by weight: MS
4.85g/L+NAA 0.3mg/L+IBA 0.3mg/L+ granularity is the activated carbon 0.5g/L+ metalaxyl 0.15mg/L+ indole fourth of 100 mesh
Acid potassium 0.3mg/L+ forchlorfenuron 0.2mg/L+ microcystin 0.04mg/L+ diethyl aminoethyl hexanoate 0.15mg/L+ putrescine 0.08mg/L, sucrose
25g/L, agar 8g/L, be adjusted to pH 5.8.
By the raw material of above-mentioned tissue culture medium (TCM) according to parts by weight, then the culture medium configured is put into high temperature sterilize pot,
131℃、1.2kg/cm2Under the conditions of carry out sterilizing 35min, then take out and be cooled to room temperature, put into and aseptic operating platform is toppled over point
Install to pass through in the culture dish of high temperature sterilize pot sterilizing.
Embodiment 2:
A kind of improving dove tree and break up the tissue culture medium (TCM) that sprouts, the raw material of described tissue culture medium (TCM) is in parts by weight: MS
8.65g/L+NAA 1.5mg/L+IBA 1.2mg/L+ granularity is the activated carbon 1.8g/L+ metalaxyl 0.20mg/L+ indole fourth of 200 mesh
Acid potassium 0.8mg/L+ forchlorfenuron 0.4mg/L+ microcystin 0.10mg/L+ diethyl aminoethyl hexanoate 0.25mg/L+ putrescine 0.15mg/L, sucrose
30g/L, agar 15g/L, be adjusted to pH 6.4.
By the raw material of above-mentioned tissue culture medium (TCM) according to parts by weight, then the culture medium configured is put into high temperature sterilize pot,
131℃、1.2kg/cm2Under the conditions of carry out sterilizing 45min, then take out and be cooled to room temperature, put into and aseptic operating platform is toppled over point
Install to pass through in the culture dish of high temperature sterilize pot sterilizing.
Embodiment 3:
A kind of improving dove tree and break up the tissue culture medium (TCM) that sprouts, described tissue culture medium (TCM) is in parts by weight: MS
5.25g/L+6-BA 1.55mg/L+NAA 0.85mg/L+IBA 0.58mg/L+ granularity is the activated carbon 0.77g/L+ first frost of 100 mesh
Spirit 0.18mg/L+ butyric acid potassium 0.55mg/L+ forchlorfenuron 0.25mg/L+ microcystin 0.06mg/L+ diethyl aminoethyl hexanoate 0.18mg/L+
Putrescine 0.12mg/L, sucrose 28g/L, agar 11g/L, be adjusted to pH 5.8.
By the raw material of above-mentioned tissue culture medium (TCM) according to parts by weight, then the culture medium configured is put into high temperature sterilize pot,
131℃、1.2kg/cm2Under the conditions of carry out sterilizing 42min, then take out and be cooled to room temperature, put into and aseptic operating platform is toppled over point
Install to pass through in the culture dish of high temperature sterilize pot sterilizing.
Embodiment 4:
Without metalaxyl+butyric acid potassium+forchlorfenuron+microcystin+diethyl aminoethyl hexanoate+putrescine, remaining is same as in Example 1.
Embodiment 5:
Without microcystin composition；Remaining is same as in Example 1.
Embodiment 6:
Microcystin content be 0.15mg/L；Remaining is same as in Example 1.
Test example: select disease-free, without incomplete dove tree leaf sections, after sterilizing, aseptically accesses embodiment
The culture medium that 1-6 prepares, each process inoculation 10 bottles, 3 every bottle, cultivation temperature (27 ± 1) DEG C, intensity of illumination
2500-3000lx, light application time 12h/d.Record sprouts the time, and bud growing state the results are shown in Table 1；
The growing state of each embodiment Seeds of Dendrobium Candidum of table 1
| Test group | Germination percentage % | Sprout the time (h) | Bud color | Bud meat thickness (cm) |
| Embodiment 1 | 100 | 40 | Light yellow-light green-green | 0.12 |
| Embodiment 2 | 100 | 41 | Light yellow-light green-green | 0.13 |
| Embodiment 3 | 100 | 38 | Light yellow-light green-green | 0.15 |
| Embodiment 4 | 97 | 62 | Light yellow-yellowish green-yellowish green | 0.09 |
| Embodiment 5 | 98 | 57 | Light yellow-yellowish green-yellowish green | 0.07 |
| Embodiment 6 | 50 | 95 | Light yellow-yellowish green-yellowish green | 0.05 |
As seen from the above table, by use the present invention technical scheme (embodiment 1-3), germination percentage reach 100%, averagely
Sprout time and bud color is also superior to other embodiments, and embodiment 3 is optimum efficiency formula.Embodiment 4 is not added first frost
Spirit+butyric acid potassium+forchlorfenuron+microcystin+diethyl aminoethyl hexanoate, other conditions are all identical with technical solution of the present invention, its germination percentage
Reach 97%, reduce 3 percentage points, and the time of sprouting relatively is added with without metalaxyl+butyric acid potassium+forchlorfenuron+microcapsule
Algin+diethyl aminoethyl hexanoate, required time to grow, do not improve seedling-growing time, and the bud color after sprouting can not keep green, and all thick
Spend thin compared with embodiment 1-3；Embodiment 5, without microcystin, other are same as in Example 1, and germination percentage reduces 2
Percentage point, the required time that sprouts is better than embodiment 4, less than embodiment 1-3, and bud color can not be kept healthy, and bud thickness is full,
Illustrate to add a composition less: microcystin, it is also possible to dove tree tissue is sprouted and has facilitation；Embodiment 6, improves micro-
The content of capsule algin, extend greatly dove tree differentiation sprout the time, also reduce dove tree bud ratio, bud embryo color,
Bud meat thickness is the most poor compared with embodiment 1-3.
Described above is the detailed description for the preferable possible embodiments of the present invention, but embodiment is not limited to the special of the present invention
Profit application range, the equal change completed under the technical spirit suggested by all present invention or modification change, the present invention all should be belonged to
Contained the scope of the claims.
Claims (6)
1. one kind is improved the tissue culture medium (TCM) that dove tree differentiation is sprouted, it is characterised in that: the raw material of described tissue culture medium (TCM) is according to weight
Part is calculated as: MS 4.85-8.65g/L+NAA 0.3-1.5mg/L+IBA 0.3-1.2mg/L+ activated carbon 0.5-1.8g/L+ metalaxyl
0.15-0.20mg/L+ butyric acid potassium 0.3-0.8mg/L+ forchlorfenuron 0.2-0.4mg/L+ microcystin 0.04-0.10mg/L+ diethyl aminoethyl hexanoate
0.15-0.25mg/L+ putrescine 0.08-0.15mg/L, sucrose 25-30g/L, agar 8-15g/L.
The most according to claim 1 a kind of improve dove tree and break up the tissue culture medium (TCM) that sprouts, it is characterised in that: described tissue
Culture medium is in parts by weight: MS 5.0-7.50g/L+NAA 0.5-1.20mg/L+IBA 0.5-0.85mg/L+ activated carbon
0.6-1.30g/L+ metalaxyl 0.17-0.19mg/L+ butyric acid potassium 0.45-0.65mg/L+ forchlorfenuron 0.22-0.35mg/L+ Microcystis aeruginosa
Element 0.05-0.09mg/L+ diethyl aminoethyl hexanoate 0.16-0.20mg/L+ putrescine 0.010-0.13mg/L, sucrose 26-19g/L, agar 8-15g/L.
The most according to claim 1 a kind of improve dove tree and break up the tissue culture medium (TCM) that sprouts, it is characterised in that: described tissue
Culture medium is in parts by weight: MS 5.25g/L+NAA 0.85mg/L+IBA 0.58mg/L+ activated carbon 0.77g/L+ metalaxyl
0.18mg/L+ butyric acid potassium 0.55mg/L+ forchlorfenuron 0.25mg/L+ microcystin 0.06mg/L+ diethyl aminoethyl hexanoate 0.18mg/L+ putrescine
0.12mg/L, sucrose 28g/L, agar 11g/L.
4. improve dove tree according to a kind of described in claim 1 or 2 or 3 and break up the tissue culture medium (TCM) sprouted, it is characterised in that:
The pH 5.8-6.4 of described tissue culture medium (TCM).
5. improve dove tree according to a kind of described in claim 1 or 2 or 3 and break up the tissue culture medium (TCM) sprouted, it is characterised in that:
The granularity of the activated carbon of described tissue culture medium (TCM) is 100-200 mesh.
6. improve dove tree according to a kind of described in claim 1 or 2 or 3 and break up the tissue culture medium (TCM) sprouted, it is characterised in that:
Described tissue culture medium (TCM) also includes that the later stage processes, be according to parts by weight by the raw material of tissue culture medium (TCM), then will configure
Culture medium puts into high temperature sterilize pot, at 131 DEG C, 1.2kg/cm2Under the conditions of carry out sterilizing 35-45min, then take out and be cooled to often
Temperature, puts into topple in aseptic operating platform and is dispensed in the culture dish passing through high temperature sterilize pot sterilizing.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111406655A (en) * | 2020-05-26 | 2020-07-14 | 陕西理工大学 | Davidia involucrata seedling and Davidia involucrata tissue culture method |
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