CN108633742A - A kind of China fir Stem tip induction culture medium and abductive approach - Google Patents
A kind of China fir Stem tip induction culture medium and abductive approach Download PDFInfo
- Publication number
- CN108633742A CN108633742A CN201810791198.8A CN201810791198A CN108633742A CN 108633742 A CN108633742 A CN 108633742A CN 201810791198 A CN201810791198 A CN 201810791198A CN 108633742 A CN108633742 A CN 108633742A
- Authority
- CN
- China
- Prior art keywords
- china fir
- culture medium
- stem tip
- tip induction
- concentration
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- 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
-
- 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 present invention relates to China fir adventitious bud tissue cultures, in particular to a kind of China fir Stem tip induction culture medium and abductive approach.A kind of composition of China fir Stem tip induction culture medium includes：DCR minimal mediums, 6 BA, NAA；A concentration of 0.78~0.82mg/L of 6 BA in the China fir Stem tip induction culture medium, a concentration of 0.28~0.32mg/L of NAA.Explant stem apex is subjected to Fiber differentiation in the culture medium of 6 BA and NAA compositions that are matched suitable for minimal medium and suitable concentration, has many advantages, such as that inductivity is high, adventitious bud is long, tissue cultures is made to have higher success rate.
Description
Technical field
The present invention relates to China fir adventitious bud field of tissue culture, in particular to a kind of China fir Stem tip induction culture medium
And abductive approach.
Background technology
The full name of tissue culture is Plant Tissue Breeding, broadly also known as cultured in vitro, refers to that institute is isolated from plant
Tissue, organ or the cell etc. needed carries out cultivating final acquisition intact plant under artificial manipulation then by sterile working
One technology.In a narrow sense refer to then being cultivated using the tissue, such as forming layer, mesophyll tissue etc. of plant various pieces
Plant is obtained, and the culture carried out by generating callus in incubation can be referred to, then passes through callus
It is differentiated to form new plant again.The China fir tissue culture technology in China develops very fast, but in the way of development inevitably
Encounter many urgent problems to be solved, this just need we carry out largely test go to solve.
Seeds one of of the China fir as fast growing, while being also our southern Major Tree Species Planteds.As afforestation is advised
The continuous expansion of mould, China fir tissue culture technology already become important nursery means.China fir tissue cultures have had been established one at present
Basic fundamental system, but the clone seeds newly selected are still needed to carry out research and experiment.
Most basic in China fir tissue culture is induction, for tissue culture industrialization, selects suitable material and hormone appropriate
Proportioning becomes the pith of induction.Hormone combinations optimize the problem that China Fir seedling shortage can be effectively relieved.But for
The kind newly selected studies deficiency to it, still needs to study different exogenous hormone processing to China fir tissue culture adventitious bud inducing
It influences, a reliable reference frame is provided for China fir tissue culture and propagation in scale.
In view of this, special propose the present invention.
Invention content
The first object of the present invention is to provide a kind of China fir Stem tip induction culture medium, for growth and material and excellent 54
Number elite tree carries out tissue cultures, and hormone combination is reasonable in the China fir Stem tip induction culture medium, has very well to China fir adventitious bud
Inducing action, have many advantages, such as that inductivity is high, adventitious bud is long.
The second object of the present invention is to provide a kind of China fir Stem tip induction method, and this method setting is reasonable, behaviour
Make simply, pollution rate is low, and induction success rate is high.
In order to realize that the above-mentioned purpose of the present invention, spy use following technical scheme：
A kind of China fir Stem tip induction culture medium, the composition of the culture medium include：DCR minimal mediums, 6-BA, NAA；Institute
State a concentration of 0.78~0.82mg/L of 6-BA in China fir Stem tip induction culture medium, a concentration of 0.28~0.32mg/L of NAA.
A kind of China fir Stem tip induction method, including：Explant sterilizing, stem apex inoculation and Fiber differentiation；The Fiber differentiation
It is cultivated using above-mentioned China fir Stem tip induction culture medium.
Growth of Chinese Fir and material and excellent No. 54 are picked out in forth generation germplasm resource bank by Fujian Yang Kou state-owned forest farms,
Using the stem apex of its current year life base portion coppice shoot as explant, in the 6-BA and NAA matched suitable for minimal medium and suitable concentration
It is cultivated in the culture medium of composition, has many advantages, such as that inductivity is high, adventitious bud is long, tissue cultures is made to have higher success rate.
A kind of China fir Stem tip induction culture medium, the composition of the culture medium include：DCR minimal mediums, 6-BA, NAA；Institute
State a concentration of 0.78~0.82mg/L of 6-BA in China fir Stem tip induction culture medium, a concentration of 0.28~0.32mg/L of NAA.
Most basic in China fir tissue culture is induction, how to realize that the first step of high yield is to obtain that height can be possessed
The tissue-cultured seedling of inductivity needs to carry out largely to test to find out optimal hormone combination combination.
DCR minimal mediums are that culture solution is commonly used in tissue culture, it can provide required nutriment such as a large amount of members for plant
Element, trace element, molysite and organic substance etc..In addition, common minimal medium also has MS, 1/2MS, 3/4MS etc., it is different
The type and content of nutriment contained by minimal medium are different, then have very for explant rudiment, bud number and budding time etc.
Big influence.Therefore, the selection of minimal medium is also highly important in tissue culture, and it is suitable to be obtained by experimental verification
The minimal medium of plant growth.
6-BA and NAA is auxin.6-BA, that is, 6- benzyl aminoadenines, alias：6-benzyl aminopurine, cell division
Element, English common name：6-Benzylaminopurine is the plant growth regulator to safety of human and livestock.It, which has, promotes cell
Division promotes undifferentiated tissue differentiation, promotes the accumulation of biological substance in vivo, the effects that promoting lateral bud, prevent aging.Just
Because of that 6-BA becomes indispensable compound in plant tissue and cell culture.Another important feature of 6-BA is
Poor mobility in plant, physiological action be confined to treatment site and its near.To consider to handle in practical applications
Method and treatment site.In the application, crop/floristics, usage and concentration, processing time and position difference, are shown
The effect or reaction come is also not quite similar, such as function and effect are poor when low concentration, and excessive concentrations may will produce adverse effect,
So needing to select suitable concentration according to without factors such as floristics, processing time and treatment sites in the application.Such as at this
Apply in different embodiments, concentration can also be 0.80mg/L etc., and result is almost the same within the scope of this.
NAA, that is, 1- methyl α-naphthyl acetates, English common name 1-naphthlcetic acid are chemistry of the people according to natural hormone
Structure, a kind of artificial synthesized broad spectrum type plant growth substance can promote cell division and expand, to promote seed to send out
Root, adventitious bud sprout, cuttage root-taking, and raising, which is bloomed, bears fruit.It is the same with other hormones, have in low concentration and growth is promoted to make
With high concentration has inhibiting effect, and different plant Different Organs are different to the susceptibility of auxin, so when in use because of basis
Floristics and suitable concentration is selected using position.Such as in the different embodiment of the application, concentration can also be
0.30mg/L etc., result is almost the same within the scope of this.
Preferably, the composition of the China fir Stem tip induction culture medium further includes：Sucrose, carragheen and activated carbon, wherein sugarcane
A concentration of 0.8~1.2g/L of a concentration of 28~32g/L of sugar, a concentration of 7~9g/L of carragheen, activated carbon.
Sucrose can provide carbon source and the energy as carbohydrate for plant, moreover it is possible to maintain the osmotic pressure of culture solution.Card
Glue i.e. carrageenan, galactolipin and Anhydrogalactose is drawn to be formed by polysaccharide sulfuric acid and 3,6- Anhydrogalactoses are straight
Chain polymerization object is formed, and coagulator use is done.Activated carbon is had an effect by suction-operated, can adsorb having in culture medium
Evil substance, the 5 hydroxymethyl furfural generated in high pressure sterilization such as phenol, quinones and the sucrose secreted in impurity, incubation
Deng.But its simultaneously may also Absorption Growth adjust element, nutrients etc., therefore suitable concentration should be found when in use.Such as at this
Apply in different embodiments, the concentration of the sucrose can also be 29g/L, 30g/L, 31g/L (result basic one within the scope of this
It causes)；The concentration of carragheen can also be 7.5g/L, 8g/L, 8.5g/L (result is almost the same within the scope of this)；The concentration of activated carbon
Can also be 0.9g/L, 1.0g/L, 1.1g/L (result is almost the same within the scope of this).
A kind of China fir Stem tip induction method, including：Explant sterilizing, stem apex inoculation and Fiber differentiation；The Fiber differentiation
The culture medium induced using above-mentioned China fir stem section is cultivated.
Preferably, the explant is stem apex of the China fir base portion coppice shoot containing bud；
Preferably, the China fir base portion coppice shoot acquisition time is June, July or November.
The induction of China fir adventitious bud is very important the selection of material.The hardness of some China firs itself is higher, it
Spray hardness compare it is more hard with other sprays, then being just not easy to induce.Older China fir explant is more had difficult labour
Raw callus, so, when choosing explant, it should be made with the current year lower base portion coppice shoot shoot of raw degree of lignification
For the material of induction.
The infection rate for the explant taken when June, July or these three month November is minimum, and fine day is selected when taking, can
Substantially reduce infection rate.
Preferably, after the sterilizing, the coppice shoot of not browning is cut into stem apex.
It is inoculated with after sterilizing, inoculation need to carry out in an aseptic environment, and inoculation is first first handled explant.It will go out
Explant after bacterium is pressed from both sides out from container with tweezers, after with knife further processing is carried out to explant：By not browning
The coppice spout is cut into stem apex.
Preferably, the explant, which sterilizes, includes：The alcohol for being 70%~75% with percent by volume impregnates 20s~50s,
Then 8~16min is impregnated with a concentration of 0.08mg/mL~0.12mg/mL mercuric chloride.
Preferably, described be seeded in gnotobasis carries out.
The many because being known as of China fir explant can be polluted, such as：China fir explant acquisition time, cleaning method, disinfection
Whether equipment, sterile environment and the consummation of the gimmick of operator etc..China fir explant sterilizes as China fir tissue culture most critical
One step, it is particularly important.
Alcohol is most common surface disinfectant, best with 70%~75% alcohol bactericidal effect, 95% or absolute alcohol
Phage surface protein matter fast dewatering can be made to solidify, form one layer of desciccator diaphragm, prevent alcohol to continue to penetrate into, bactericidal effect is big
It is big to reduce.Alcohol has stronger penetration power, so that bacterium protein is denaturalized, bactericidal effect is good.But alcohol kills vegetable material
Wound effect is also very big, and the growth of long soaking time, vegetable material will be affected, or even be killed by alcohol, and when use answers
Stringent control time.But alcohol is unable to thorough disinfection, is not used alone generally, is mostly used cooperatively with other disinfectants.Mercuric chloride is
Mercury chloride is white crystal, particle or powder, can inhibit the activity of bacterium enzyme containing sulfydryl, makes bacterial metabolism obstacle, can also be with egg
It binds directly in vain, makes albuminous degeneration.Mercuric chloride common a concentration of 0.1%, i.e. 0.1mg/mL.It such as in various embodiments, can
To be that 70% alcohol impregnates 25s with percent by volume, 0.1% mercuric chloride impregnates 10min, and 75% alcohol impregnates 40s, 0.1% mercuric chloride
Impregnate 14min etc..
The tissue-cultured seedling being inoculated with, once having spent this period, can generally be polluted in the 7th to the 14th infection rate highest
Rate will drastically decline.The reason of pollution, is roughly divided into two kinds of culture medium pollution and material contamination.Most culture medium pollution
All be when inoculation with caused by extraneous contact, seldom partially due to when high-temperature sterilization does not have sterilizing in place caused,
Therefore, it in inoculation, has to ensure the sterile of workbench, to improve inductivity.And the reason of material contamination, is then complex,
It is not thorough enough when may be disinfection, it is also possible to long or to be exposed in air too long very much with blade contact when being inoculated with, it is also possible to
Bottle cap pollution etc..In order to reduce pollution rate, inductivity, the completion for needing each step all extreme cares careful are improved.
Preferably, for the stem apex to be linked into the culture medium, the stem apex length of access is 2.0 for the inoculation
~3.0cm.
Inoculation is that sterilized good explant segment or fritter are put into the process of culture medium.
Preferably, the temperature of the culture be 24~26 DEG C, intensity of illumination be 1500~2000lx, light application time be 12~
14h/d。
Culture refers to that culture materials are placed on culturing room's (having illumination, temperature condition) is inner, is allowed to grow, the mistake of Differentiation
Journey.The suitable cultivation temperature of different plants is different with intensity of illumination and time.As in various embodiments, the condition of culture is also
Can be temperature it be 25 DEG C, intensity of illumination 1800lx, light application time 13h/d；Temperature is 26 DEG C, and intensity of illumination is
2000lx, light application time 12h/d；Temperature is 24 DEG C, intensity of illumination 1600lx, and light application time is 14h/d etc..
Preferably, the China fir is No. 54 trees in the forth generation germplasm resource bank of Fujian Yang Kou state-owned forest farms.
No. 54 trees are growth and material and excellent elite tree.
Compared with prior art, beneficial effects of the present invention are：
The molding of (1) species depends primarily on genotypic and environment, in the present solution, excellent Clones of Cunninghamia Lanceolata generation
Table genotype, and the culture medium of the China fir stem section induction of different hormone combinations is then environment, and the two is combined with each other, final energy
The preferred plan that best environmental condition has enough been obtained to be induced as China fir explant.
(2) the application has obtained the concentration of suitable 6-BA and NAA by many experiments and data analysis, in this range
It is interior, the induction of bud can be effectively facilitated, and excessively there will be inhibiting effect.
(3) the application is also obtained in medium component by orthogonal experiment and data statistic analysis to No. 54 elite tree stems
Sharp inductivity influence sequence be：6-BA>NAA>Minimal medium, that is to say, that 6-BA is No. 54 elite tree Stem tip induction rates
Leading factor；Having obtained the influence in medium component to No. 54 elite tree stem apex bud length sequentially is：6-BA>NAA>Basic culture
The leading factor of base, stem apex bud length is 6-BA.
(4) Growth of Chinese Fir and material of the application to be picked out in the forth generation germplasm resource bank of Fujian Yang Kou state-owned forest farms
And No. 54 excellent elite trees are material, and the processing of various concentration exogenous hormone has been obtained to China fir by many experiments and data analysis
The influence of tissue culture Stem tip induction provides a reliable reference frame for China fir tissue culture, has social benefit and scientific research value.
Specific implementation mode
Embodiment of the present invention is described in detail below in conjunction with embodiment, but those skilled in the art will
Understand, the following example is merely to illustrate the present invention, and is not construed as limiting the scope of the invention.It is not specified in embodiment specific
Condition person carries out according to conventional conditions or manufacturer's recommended conditions.Reagents or instruments used without specified manufacturer is
It can be with conventional products that are commercially available.
In the application specific implementation mode, the calculation and meaning of inductivity, average bud length and range analysis are as follows：
K2=at DCR or 0.8mg/l6-BA or 0.2mg/lNAA everywhere in reason the sum of result
Induce number：Successfully induce the bottle number of adventitious bud；
Inoculation number：The bottle number being inoculated with.
It is bigger, it is bigger on test index influence, reflect the influence of each small factors on test indicators in three main factors
Size.
R：It is bigger, it is bigger on test index influence, it is more important, it can reflect the primary-slave relation of three main factors.
MS used in this application is shown in Table 1 and table respectively with DCR minimal medium mother liquor formulas and auxin mother liquor formula
2。
1 MS of table and DCR mother liquor formula tables
2 auxin mother liquor formula table of table
Embodiment 1
In the morning of a certain fair weather in June, chooses in the forth generation germplasm resource bank of Fujian Yang Kou state-owned forest farms and select
The growth gone out is explant with the stem apex of the base portion coppice shoot containing bud of material and excellent No. 54 elite trees of China fir, removes extra blade,
Flowing water uses sterile water wash 2 times after rinsing well, impregnates 25s with 70% alcohol, then cleaned 2 times with asepsis, then uses
0.1% mercuric chloride impregnates 10min, with sterile water wash 5 times.The explant that sterilizing is completed is placed on spare in vial.
China fir Stem tip induction culture medium is prepared, and required article is put into workbench and is sterilized 30 minutes with ultraviolet lamp, behaviour
Work person's handle is sterilized with alcohol swab, and tweezers and knife are impregnated with alcohol, after the calcination on alcolhol burner, by explant tweezers from glass
Pressed from both sides out in glass bottle, after with knife further processing is carried out to explant：Stem apex is switched to no browning part.It will be located with tweezers again
The stem apex managed is inserted into culture bottle, and the length of access is about 2cm, after bottle cap is twisted (bottle cap needs the first calcination piece under alcolhol burner
It carves).Tweezers and scalpel are required for being sterilized with alcolhol burner after being inoculated with every time.In the enterprising rower of culture medium after being all inoculated with
Remember, the selected blaze of each bottle, inoculation date and inoculation people in note, in order to record data.
The culture medium being inoculated with is put into culturing room to cultivate, culture room temperature is maintained at 25 DEG C or so, intensity of illumination
1800lx, light application time 13h/d.
It is recorded every three days to the pollution number of five days docking seedlings, stem apex bud length.Every group of experiment is 3 times parallel.To data
Carry out range analysis.
Embodiment 2
In the morning of a certain fair weather in July, chooses in the forth generation germplasm resource bank of Fujian Yang Kou state-owned forest farms and select
The growth gone out is explant with stem apex of the base portion coppice shoot containing bud of material and excellent China fir 54, removes extra blade, flowing water punching
Sterile water wash is used after wash clean 2 times, 40s is impregnated with 75% alcohol, then cleaned 2 times with asepsis, then with 0.11% liter
Mercury impregnates 14min, with sterile water wash 5 times.The explant that sterilizing is completed is placed on spare in vial.
China fir Stem tip induction culture medium is prepared, and required article is put into workbench and is sterilized 30 minutes with ultraviolet lamp, behaviour
Work person's handle is sterilized with alcohol swab, and tweezers and knife are impregnated with alcohol, after the calcination on alcolhol burner, by explant tweezers from glass
Pressed from both sides out in glass bottle, after with knife further processing is carried out to explant：Stem apex is switched to no browning part.It will be located with tweezers again
The stem apex managed is inserted into culture bottle, and the length of access is about 2.5cm, after bottle cap is twisted (bottle cap needs the first calcination under alcolhol burner
For a moment).Tweezers and scalpel are required for being sterilized with alcolhol burner after being inoculated with every time.It is carried out on culture medium after being all inoculated with
It marks, the selected blaze of each bottle, inoculation date and inoculation people in note, in order to record data.
The culture medium being inoculated with is put into culturing room to cultivate, culture room temperature is maintained at 26 DEG C or so, intensity of illumination
2000lx, light application time 12h/d.
It is recorded every three days to the pollution number of five days docking seedlings, stem apex bud length.Every group of experiment is 3 times parallel.To data
Carry out range analysis.
Embodiment 3
In the morning of a certain fair weather in November, chooses in the forth generation germplasm resource bank of Fujian Yang Kou state-owned forest farms and select
The growth gone out is explant with the stem apex of the base portion coppice shoot containing bud of material and excellent No. 54 elite trees of China fir, removes extra blade,
Flowing water uses sterile water wash 2 times after rinsing well, impregnates 40s with 75% alcohol, then cleaned 2 times with asepsis, then uses
0.11% mercuric chloride impregnates 14min, with sterile water wash 5 times.The explant that sterilizing is completed is placed on spare in vial.
China fir Stem tip induction culture medium is prepared, and required article is put into workbench and is sterilized 30 minutes with ultraviolet lamp, behaviour
Work person's handle is sterilized with alcohol swab, and tweezers and knife are impregnated with alcohol, after the calcination on alcolhol burner, by explant tweezers from glass
Pressed from both sides out in glass bottle, after with knife further processing is carried out to explant：Stem apex is switched to no browning part.It will be located with tweezers again
The stem apex managed is inserted into culture bottle, and the length of access is about 2.8cm, after bottle cap is twisted (bottle cap needs the first calcination under alcolhol burner
For a moment).Tweezers and scalpel are required for being sterilized with alcolhol burner after being inoculated with every time.It is carried out on culture medium after being all inoculated with
It marks, the selected blaze of each bottle, inoculation date and inoculation people in note, in order to record data.
The culture medium being inoculated with is put into culturing room to cultivate, culture room temperature is maintained at 24 DEG C or so, intensity of illumination
1600lx, light application time 14h/d.
It is recorded every three days to the pollution number of five days docking seedlings, stem apex bud length.Every group of experiment is 3 times parallel.To data
Carry out range analysis.
Experimental example 1
The influence of the pairs of inductivity of different China fir Stem tip induction culture medium groups.
Based on embodiment 3, it is related to the change of three experiment parameters：Minimal medium, 6-BA, NAA, its in culture medium
His ingredient such as sucrose, carragheen and activated carbon additive amount are constant, and experimental method is the same as embodiment 3.
Wherein minimal medium is divided into 1/2MS, MS and DCR, and the concentration of 6-BA is divided into the concentration point of 0.6,0.8,1.2, NAA
It is 0.1,0.2,0.3, specific proportioning is as shown in the table：
3 not isogeneous induction China fir Stem tip induction culture medium L9 (3 of table3) formula orthogonal test table
Each China fir Stem tip induction culture medium is a processing, each 30 bottles of processing inoculation, one explant of every bottle of inoculation
Body, experiment are repeated 3 times.
It is for statistical analysis to experimental data, it the results are shown in Table 4.
The impact analysis table of the pairs of inductivity of the different China fir Stem tip induction culture medium groups of table 4
From table 4, it can be seen that No. 54 elite tree stem apexs of various concentration pair of variety classes minimal medium and 6-BA, NAA
Inductivity have a great impact.The numerical values recited of inductivity is intuitively seen first, then No. 54 elite tree stem apexs are in No. 7 culture mediums
That is the inductivity highest under 3/4MS+0.6mg/L 6-BA+0.3mg/L NAA, reaches 72.2%；But byValue is it is found that basic training
The optimal level for supporting base is the second horizontal DCR, and the optimal level of 6-BA is the second horizontal 0.8mg/L, and the optimal level of NAA is the
Three horizontal 0.3mg/L, therefore, theoretically the optimal medium of No. 54 elite tree Stem tip induction rates is DCR+0.8mg/L 6-BA+
0.3mg/L NAA (processing 5；Inductivity 68.8% is close with 72.2%)；According to R values, by being ranked sequentially from big to small：6-BA
>NAA>Minimal medium, therefore, 6-BA are the leading factor for influencing No. 54 elite tree Stem tip induction rates.But practical and theoretical
Culture medium is inconsistent, and by independent experiment, the contrast test that repeats twice that two kinds of processing are arranged finds that processing 5 is repeated several times
Average inductivity in experiment is higher than the average inductivity (76.7% for handling 7>73.4%).Therefore, it is conducive to No. 54 elite tree stems
The optimal medium of point induction is DCR+0.8mg/L 6-BA+0.3mg/L NAA.
It is substantially consistent with embodiment 3 that embodiment 1 and embodiment 2 handle lower result herein.
Experimental example 2
The averagely influence of bud length in pairs of different China fir Stem tip induction culture medium groups.
Experimental setup and method are the same as experimental example 1.Since institute all induces into either with or without contaminated China fir stem apex in an experiment
Work(, thus selected the 15th day as observation stem apex be averaged bud grow time.It in this way can be to avoid when seriously polluted
Misregister is waited, can also avoid wait until that the later stage most of all grow the similar time-division and do not go out quality.
It is for statistical analysis to experimental data, it the results are shown in Table 5.
The different China fir Stem tip induction culture medium groups of table 5 analytical table that averagely bud length influences in pairs
As can be seen from Table 5, No. 54 elite tree stem apexs of various concentration pair of variety classes minimal medium and 6-BA, NAA
The average bud length of growth has a great impact.It intuitively can be seen that stem apex is using No. 2 i.e. 1/2MS+0.8mg/L of culture medium first
Bud longest under 6-BA+0.2mg/L NAA, reaches 4.8cm；ByValue is it is found that the optimal level of minimal medium is the first water
The optimal level of flat 1/2MS, 6-BA are the second horizontal 0.8mg/L, and the optimal level of NAA is the second horizontal 0.2mg/L, therefore,
Theoretically the optimal medium of No. 54 elite tree stem apex buds length is 1/2MS+0.8mg/L6-BA+0.2mg/L NAA；According to R values,
By being ranked sequentially from big to small：6-BA>NAA>Minimal medium, therefore, 6-BA are to influence No. 54 clone stem apex buds to grow
Leading factor.But experimental result, which is repeated several times, to be confirmed, the bud length of DCR+0.8mg/L 6-BA+0.3mg/L NAA also reaches
4.5cm.Therefore, in Binding experiment example 1 the pairs of inductivity of different culture media group influence, obtain No. 54 elite tree stem apex buds length length
It is No. 5 culture mediums i.e. DCR+0.8mg/L6-BA+0.3mg/L NAA to spend maximum optimal medium.
It is substantially consistent with embodiment 3 that embodiment 1 and embodiment 2 handle lower result herein.
Finally it should be noted that：The above embodiments are only used to illustrate the technical solution of the present invention., rather than its limitations；To the greatest extent
Present invention has been described in detail with reference to the aforementioned embodiments for pipe, but it will be understood by those of ordinary skill in the art that：Its
It still can be with technical scheme described in the above embodiments is modified, either to which part or all technical features
Carry out equivalent replacement；And these modifications or replacements, various embodiments of the present invention skill that it does not separate the essence of the corresponding technical solution
The range of art scheme.
Claims (10)
1. a kind of China fir Stem tip induction culture medium, which is characterized in that the composition of the culture medium includes：DCR minimal mediums, 6-
BA、NAA；
A concentration of the 0.28 of a concentration of 0.78~0.82mg/L of 6-BA in the China fir Stem tip induction culture medium, NAA~
0.32mg/L。
2. China fir Stem tip induction culture medium according to claim 1, which is characterized in that the China fir Stem tip induction culture medium
Composition further include：Sucrose, carragheen and activated carbon, wherein a concentration of 28~32g/L of sucrose, carragheen a concentration of 7~
A concentration of 0.8~1.2g/L of 9g/L, activated carbon.
3. a kind of China fir Stem tip induction method, which is characterized in that including：Explant sterilizing, stem apex inoculation and Fiber differentiation；
The Fiber differentiation is cultivated using the China fir Stem tip induction culture medium described in claim 2.
4. China fir Stem tip induction method according to claim 3, which is characterized in that the explant is China fir base portion coppice shoot
Stem apex；
Preferably, the China fir base portion coppice shoot acquisition time is June, July or November.
5. China fir Stem tip induction method according to claim 4, which is characterized in that after the sterilizing, will there is no browning
The coppice shoot be cut into stem apex.
6. China fir Stem tip induction method according to claim 3, which is characterized in that the explant, which sterilizes, includes：Use body
The alcohol that product percentage is 70%~75% impregnates 20s~50s, is then soaked with a concentration of 0.08mg/mL~0.12mg/mL mercuric chloride
Steep 8~16min.
7. China fir Stem tip induction method according to claim 3, which is characterized in that it is described be seeded in gnotobasis into
Row.
8. China fir Stem tip induction method according to claim 3, which is characterized in that the inoculation is to access the stem apex
Into the culture medium, the stem apex length of access is 2.0~3.0cm.
9. China fir Stem tip induction method according to claim 3, which is characterized in that the temperature of the culture is 24~26
DEG C, intensity of illumination is 1500~2000lx, and light application time is 12~14h/d.
10. according to claim 3~9 any one of them China fir Stem tip induction method, which is characterized in that the China fir is Fujian
No. 54 trees in the forth generation germplasm resource bank of foreign mouth state-owned forest farms.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810791198.8A CN108633742A (en) | 2018-07-18 | 2018-07-18 | A kind of China fir Stem tip induction culture medium and abductive approach |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810791198.8A CN108633742A (en) | 2018-07-18 | 2018-07-18 | A kind of China fir Stem tip induction culture medium and abductive approach |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108633742A true CN108633742A (en) | 2018-10-12 |
Family
ID=63759729
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810791198.8A Pending CN108633742A (en) | 2018-07-18 | 2018-07-18 | A kind of China fir Stem tip induction culture medium and abductive approach |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108633742A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108575763A (en) * | 2018-07-20 | 2018-09-28 | 南京林业大学 | A kind of China fir stem section inducing culture and abductive approach |
| CN109479713A (en) * | 2018-12-03 | 2019-03-19 | 南京林业大学 | A kind of coppice spout using China fir carries out root induction method as explant |
-
2018
- 2018-07-18 CN CN201810791198.8A patent/CN108633742A/en active Pending
Non-Patent Citations (2)
| Title |
|---|
| 席梦利等: "杉木遗传转化受体系统的建立", 《南京林业大学学报（自然科学版）》 * |
| 朱立新等: "《园艺通论》", 30 April 2015 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108575763A (en) * | 2018-07-20 | 2018-09-28 | 南京林业大学 | A kind of China fir stem section inducing culture and abductive approach |
| CN109479713A (en) * | 2018-12-03 | 2019-03-19 | 南京林业大学 | A kind of coppice spout using China fir carries out root induction method as explant |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Taji et al. | Plant tissue culture practice | |
| KR20140040513A (en) | Production of virus free plants from in vitro shoot tips through in vitro meristem culture | |
| CN105706880A (en) | Purple sweet potato virus-free seedling water culture fast breeding method | |
| Rafiq et al. | Standardization of in vitro micropropagation procedure of Oriental Lilium Hybrid Cv.‘Ravenna’ | |
| CN106982737A (en) | The regeneration culture medium and cultural method of blueberry tissue culture and application | |
| CN108633742A (en) | A kind of China fir Stem tip induction culture medium and abductive approach | |
| CN106942053A (en) | A kind of tissue culture and rapid propagation method for Xingan lucid asparagus | |
| CN109042330A (en) | A kind of method for tissue culture of spindle tree | |
| CN102232359B (en) | In-vitro rapid propagation method of double-petal Jasminum sambac | |
| CN113099866B (en) | Method for rapidly propagating fir excellent family seedlings | |
| CN102630564A (en) | Tissue culture and rapid propagation method of salt-tolerant field mint | |
| KR101887221B1 (en) | Method of mass propagation of bamboo by in vitro culture | |
| CN108575763A (en) | A kind of China fir stem section inducing culture and abductive approach | |
| Llorente et al. | In vitro propagation of fraser photinia using Azospirillum-mediated root development | |
| CN109496706A (en) | A kind of sterile brachyplast of Mao Xu Cao quickly breed before explant preprocess method | |
| CN105684910B (en) | The tissue culture and rapid propagation method of dendrobium hancockii Rolfe | |
| CN114946654A (en) | Phalaenopsis stem tip growth point culture virus removal technology | |
| CN105230493B (en) | A kind of propagation method of bighead atractylodes rhizome seedling and its application | |
| Zemene et al. | Protocol optimization for micro-propagation of Green pepper (Capsicum annum L.) cultivated in Ethiopia | |
| CN107410019A (en) | A kind of method that sweet potato tissue cultures are carried out using classification culture medium | |
| Zhai et al. | Shoot multiplication and plant regeneration in Caragana fruticosa (Pall.) Besser | |
| CN112602597A (en) | Method for obtaining industrial hemp full-female or full-male seedlings | |
| CN106305419B (en) | A kind of method of open culture sugar-cane tissue culture seedlings | |
| Singh et al. | Plant tissue culture | |
| CN108849511A (en) | A kind of method for tissue culture of the new poplar seedling of hair |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20181012 |