CN107151181A - A kind of Tissue-cultured apple seedling culture medium and preparation method thereof - Google Patents
A kind of Tissue-cultured apple seedling culture medium and preparation method thereof Download PDFInfo
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- CN107151181A CN107151181A CN201710547939.3A CN201710547939A CN107151181A CN 107151181 A CN107151181 A CN 107151181A CN 201710547939 A CN201710547939 A CN 201710547939A CN 107151181 A CN107151181 A CN 107151181A
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- culture medium
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05B—PHOSPHATIC FERTILISERS
- C05B7/00—Fertilisers based essentially on alkali or ammonium orthophosphates
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
Abstract
The invention discloses a kind of Tissue-cultured apple seedling culture medium and preparation method thereof, belong to Apple Culture technical field, be made up of the raw material of following parts by weight：20 30 parts of ammonium sulfate, 15 25 parts of sodium nitrate, 10 15 parts of potassium dihydrogen phosphate, 13 parts of ferrous sulfate, 0.5 3 parts of zinc sulfate, 80 100 parts of agar, 0.5 1 parts of the basic element of cell division, 12 parts of heteroauxin, 0.5 1 parts of 6 benayl aminopurine, 100 200 parts of sterilized water, 20 30 parts of glucose, viral 7 18 parts of ether, 8 20 parts of sodium hypochlorite.The culture medium comprehensive nutrition, makes that tissue culture seedling rooting is rapid, survival rate is high, substitutes high-temperature sterilization by adding sodium hypochlorite in culture medium, carries out detoxification by adding viral ether, sterilizing, detoxification efficiency are good.
Description
Technical field
The invention belongs to Apple Culture technical field, and in particular to a kind of Tissue-cultured apple seedling culture medium and preparation method thereof.
Background technology
Apple is perennial woody plant, and the tissue cultures of apple are to use Sterile Culture Methods Used, will come from good plant
The organ such as stem apex, axillary bud, blade and their histotomy carry out cultured in vitro, be allowed to obtain a large amount of heredity in a short time
Property consistent individual method, with the cycle it is short, be easy to manually control condition of culture, reproduction speed soon, occupy little space, economy
The advantages of high efficiency, meanwhile, also accelerate the popularization and application of Variety of Apple.Culture medium is the important material of Plant Tissue Breeding
Material, is the nutriment of explant growth, only makes suitable culture medium, can just tissue cultures is succeeded.
Sterilization technology is one of key technology in tissue culture procedures, at present, the medium sterilization of Plant Tissue Breeding
Method is to use high pressure steam sterilization method, there is following shortcoming：Equipment is more compared with expensive, the consumption energy, the training of sterilization time length
Some chemical reactions can be occurred by supporting in base, culture medium into branch along with occurring some changes.In addition, the apple of China's plant
A variety of viruses are generally infected, apple mosaic virus, apple chlorotic leaf spot virus, apple stem pitting virus and apple stem grooving virus are to invade
The most common virus of apple is contaminated, Apple virus disease has a strong impact on the quality and yield of fruit, and apple once infects virus, is difficult to
Preventing and treating.
The content of the invention
The present invention provides a kind of Tissue-cultured apple seedling culture medium and preparation method thereof, and the culture medium comprehensive nutrition makes tissue-cultured seedling
Rapid, survival rate of taking root is high, and high-temperature sterilization is substituted by adding sodium hypochlorite in culture medium, is taken off by adding viral ether
Poison, sterilizing, detoxification efficiency are good.
The present invention uses following technical scheme：
A kind of Tissue-cultured apple seedling culture medium, is made up of the raw material of following parts by weight：20-30 parts of ammonium sulfate, sodium nitrate 15-25
Part, 10-15 parts of potassium dihydrogen phosphate, 1-3 parts of ferrous sulfate, 0.5-3 parts of zinc sulfate, 80-100 parts of agar, basic element of cell division 0.5-1
Part, 1-2 parts of heteroauxin, 0.5-1 parts of 6-benzyl aminopurine, 100-200 parts of sterilized water, 20-30 parts of glucose, viral ether 7-18
Part, 8-20 parts of sodium hypochlorite.
It is preferred that, it is made up of the raw material of following parts by weight：22-26 parts of ammonium sulfate, 18-22 parts of sodium nitrate, potassium dihydrogen phosphate
12-15 parts, 1-2 parts of ferrous sulfate, 1-2 parts of zinc sulfate, 90-100 parts of agar, 0.6-0.8 parts of the basic element of cell division, heteroauxin 1-
1.5 parts, 0.5-0.8 parts of 6-benzyl aminopurine, 150-200 parts of sterilized water, 20-25 parts of glucose, viral 10-16 parts of ether, secondary chlorine
Sour sodium 10-14 parts.
It is preferred that, it is made up of the raw material of following parts by weight：24 parts of ammonium sulfate, 20 parts of sodium nitrate, 13 parts of potassium dihydrogen phosphate, sulphur
Sour ferrous 1.5 parts, 1 part of zinc sulfate, 100 parts of agar, 0.6 part of the basic element of cell division, 1 part of heteroauxin, 6-benzyl aminopurine 0.6
Part, 180 parts of sterilized water, 22 parts of glucose, viral 13 parts of ether, 12 parts of sodium hypochlorite.
Present invention also offers the preparation method of the Tissue-cultured apple seedling culture medium, comprise the following steps：According to weight percent
Than weighing various raw materials, heating stirring until whole be uniformly dissolved obtains culture medium mother liquor, cooling, regulation mother liquor pH value is 5.8-
6.0, finally sealed preserves and produces the culture medium.
Present invention additionally comprises the application of the Tissue-cultured apple seedling culture medium in apple tissue culture.
The present invention advantageous effects be：Using the culture medium of reasonable component, comprehensive nutrition, the tissue culture seedling rooting of culture
Rapidly, survival rate is high, improves the speed and efficiency of tissue culture；High-temperature sterilization is substituted by sodium hypochlorite, without carrying out the culture
The high-temperature sterilization of base, can avoid high-temperature sterilization to the adverse effect of nutritional ingredient in the culture medium, be conducive to plant growth；It is logical
The viral ether for crossing addition low dosage in the medium realizes the purpose of detoxification, and detoxification efficiency is good, smaller to plant Survival Effects.
Embodiment
With reference to specific embodiment, the invention will be further described.
Embodiment 1
A kind of Tissue-cultured apple seedling culture medium, is made up of the raw material of following parts by weight：24 parts of ammonium sulfate, 20 parts of sodium nitrate, phosphorus
13 parts of acid dihydride potassium, 1.5 parts of ferrous sulfate, 1 part of zinc sulfate, 100 parts of agar, 0.6 part of the basic element of cell division, 1 part of heteroauxin, 6-
0.6 part of benayl aminopurine, 180 parts of sterilized water, 22 parts of glucose, viral 13 parts of ether, 12 parts of sodium hypochlorite.According to percentage by weight
Weigh various raw materials, heating stirring and obtain culture medium mother liquor until being all uniformly dissolved, cool down, regulation mother liquor pH value is 5.8-
6.0, finally sealed preserves and produces the culture medium.
Embodiment 2
A kind of Tissue-cultured apple seedling culture medium, is made up of the raw material of following parts by weight：22 parts of ammonium sulfate, 18 parts of sodium nitrate, phosphorus
12 parts of acid dihydride potassium, 1 part of ferrous sulfate, 1 part of zinc sulfate, 90 parts of agar, 0.7 part of the basic element of cell division, 1.2 parts of heteroauxin, 6-
0.5 part of benayl aminopurine, 160 parts of sterilized water, 20 parts of glucose, viral 12 parts of ether, 14 parts of sodium hypochlorite.According to percentage by weight
Weigh various raw materials, heating stirring and obtain culture medium mother liquor until being all uniformly dissolved, cool down, regulation mother liquor pH value is 5.8-
6.0, finally sealed preserves and produces the culture medium.
Embodiment 3
A kind of Tissue-cultured apple seedling culture medium, is made up of the raw material of following parts by weight：26 parts of ammonium sulfate, 22 parts of sodium nitrate, phosphorus
15 parts of acid dihydride potassium, 2 parts of ferrous sulfate, 2 parts of zinc sulfate, 100 parts of agar, 0.8 part of the basic element of cell division, 1.5 parts of heteroauxin, 6-
0.8 part of benayl aminopurine, 200 parts of sterilized water, 25 parts of glucose, viral 16 parts of ether, 10 parts of sodium hypochlorite.According to percentage by weight
Weigh various raw materials, heating stirring and obtain culture medium mother liquor until being all uniformly dissolved, cool down, regulation mother liquor pH value is 5.8-
6.0, finally sealed preserves and produces the culture medium.
Beneficial effects of the present invention are further illustrated below by way of experiment：
Table 1 be the conventional apple culture medium of embodiment 1-3 and control group 1 (10 parts of peptone, 3 parts of beef extract, 5 parts of sodium chloride,
15-20 parts of agar, 1000 parts of distilled water) result of the test taken root to Tissue-cultured apple seedling.
Table 1
| Processing | Rooting rate/% | Root length/cm | Radical | Root thickness |
| Embodiment 1 | 97.2 | 1.75 | 9.10 | It is medium |
| Embodiment 2 | 98.3 | 1.76 | 9.20 | It is thicker |
| Embodiment 3 | 98.16 | 1.72 | 9.14 | It is thicker |
| Control group 1 | 86 | 1.63 | 8.2 | It is thinner |
Table 1 shows that the tissue culture medium (TCM) of the present invention is more preferable relative to the effect that conventional medium is taken root to Tissue-cultured apple seedling.
When Tissue-cultured apple seedling expand in culture medium it is numerous to requirement after, the consistent plant of growing way is chosen, using RT-PCR
Method to embodiment 1-3 with the plant for the medium controls 2 for being not added with viral ether virus detect, calculating detoxification
Effect, as shown in table 2.
Table 2
| Processing | Handle strain number | Cut stem apex number | Survive stem apex number/(%) | Avirulent strain tree/(%) |
| Embodiment 1 | 60 | 178 | 157(88.2) | 130(82.8) |
| Embodiment 2 | 60 | 210 | 194(92.4) | 168(86.6) |
| Embodiment 3 | 60 | 192 | 181(94.3) | 166(91.7) |
| Control group 2 | 60 | 180 | 142(78.9) | 102(71.8) |
From result above, by adding viral ether in the medium, ensureing Tissue-cultured apple seedling compared with high-survival rate
Under the conditions of, viral removal efficiency is high, meets the demand of Apple virus removing.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all skills in the present invention
On the basis of art scheme, those skilled in the art need not pay the various modifications that creative work can make or deformation still exists
In protection scope of the present invention.
Claims (6)
1. a kind of Tissue-cultured apple seedling culture medium, it is characterised in that be made up of the raw material of following parts by weight：20-30 parts of ammonium sulfate, nitre
Sour sodium 15-25 parts, 10-15 parts of potassium dihydrogen phosphate, 1-3 parts of ferrous sulfate, 0.5-3 parts of zinc sulfate, 80-100 parts of agar, cell point
Split plain 0.5-1 parts, 1-2 parts of heteroauxin, 0.5-1 parts of 6-benzyl aminopurine, 100-200 parts of sterilized water, 20-30 parts of glucose,
Viral ether 7-18 parts, 8-20 parts of sodium hypochlorite.
2. a kind of Tissue-cultured apple seedling culture medium according to claim 1, it is characterised in that by the raw material group of following parts by weight
Into：22-26 parts of ammonium sulfate, 18-22 parts of sodium nitrate, 12-15 parts of potassium dihydrogen phosphate, 1-2 parts of ferrous sulfate, 1-2 parts of zinc sulfate, fine jade
90-100 parts of fat, 0.6-0.8 parts of the basic element of cell division, 1-1.5 parts of heteroauxin, 0.5-0.8 parts of 6-benzyl aminopurine, sterilized water
150-200 parts, 20-25 parts of glucose, viral 10-16 parts of ether, 10-14 parts of sodium hypochlorite.
3. a kind of Tissue-cultured apple seedling culture medium according to claim 1, it is characterised in that by the raw material group of following parts by weight
Into：It is 24 parts of ammonium sulfate, 20 parts of sodium nitrate, 13 parts of potassium dihydrogen phosphate, 1.5 parts of ferrous sulfate, 1 part of zinc sulfate, 100 parts of agar, thin
0.6 part of born of the same parents' mitogen, 1 part of heteroauxin, 0.6 part of 6-benzyl aminopurine, 180 parts of sterilized water, 22 parts of glucose, viral ether 13
Part, 12 parts of sodium hypochlorite.
4. a kind of preparation method of Tissue-cultured apple seedling culture medium described in claim any one of 1-3, it is characterised in that including with
Lower step：Various raw materials, heating stirring, which are weighed, according to percentage by weight obtains culture medium mother liquor until being all uniformly dissolved, it is cold
But, mother liquor pH value is adjusted, finally sealed preserves and produces the culture medium.
5. a kind of preparation method of Tissue-cultured apple seedling culture medium according to claim 4, it is characterised in that the mother liquor pH
It is worth for 5.8-6.0.
6. application of a kind of Tissue-cultured apple seedling culture medium in apple tissue culture described in claim any one of 1-3.
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| CN201710547939.3A CN107151181A (en) | 2017-07-06 | 2017-07-06 | A kind of Tissue-cultured apple seedling culture medium and preparation method thereof |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110169420A (en) * | 2019-05-24 | 2019-08-27 | 京博农化科技有限公司 | A kind of composition pesticide that heteroauxin is compounded with ribavirin and application |
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| CN104429942A (en) * | 2014-11-03 | 2015-03-25 | 中国农业科学院果树研究所 | Detoxification method combining heat treatment and chemical treatment for apple tissue culture seedlings |
| CN105284627A (en) * | 2015-12-07 | 2016-02-03 | 山东省烟台市农业科学研究院 | Method for removing Fuji apple tissue cultured seedling rust fruit-like viruses |
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| CN104186313A (en) * | 2014-08-04 | 2014-12-10 | 江苏农林职业技术学院 | Inducing culture medium of malus domestica borkh pulp callus tissue and inducing multiplication culture method of inducing culture medium |
| CN104429942A (en) * | 2014-11-03 | 2015-03-25 | 中国农业科学院果树研究所 | Detoxification method combining heat treatment and chemical treatment for apple tissue culture seedlings |
| CN105284627A (en) * | 2015-12-07 | 2016-02-03 | 山东省烟台市农业科学研究院 | Method for removing Fuji apple tissue cultured seedling rust fruit-like viruses |
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| CN110169420A (en) * | 2019-05-24 | 2019-08-27 | 京博农化科技有限公司 | A kind of composition pesticide that heteroauxin is compounded with ribavirin and application |
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