CN111903529A - Rapid propagation technology for apple tissue culture seedlings - Google Patents
Rapid propagation technology for apple tissue culture seedlings Download PDFInfo
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- CN111903529A CN111903529A CN202010977539.8A CN202010977539A CN111903529A CN 111903529 A CN111903529 A CN 111903529A CN 202010977539 A CN202010977539 A CN 202010977539A CN 111903529 A CN111903529 A CN 111903529A
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- tissue culture
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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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- 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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Abstract
The invention provides a rapid propagation method of tissue culture seedlings of apples, which comprises the following steps: culturing the apple tissue culture seedlings by using a subculture multiplication culture medium for 30-40 days, then transferring the apple tissue culture seedlings to a rooting culture medium for culturing for 25-35 days, and finally transplanting the rooted tissue culture seedlings to a transplanting substrate by using a nutrition pot. When the multiplication medium provided by the invention is used, the multiplication coefficient averagely reaches 6.1, which is improved by 8.9% compared with the prior art; the rooting culture medium provided by the invention achieves the effect of rooting, remarkably improves the rooting rate of the tissue culture seedling, achieves 85 percent on average and is improved by 6.25 percent compared with the prior art; the survival rate of the tissue culture seedlings is improved by the transplanting matrix provided by the invention to over 75 percent and up to 85 percent, and is improved by 6.25 to 7.14 percent compared with the prior art.
Description
Technical Field
The invention relates to the technical field of apple seedling breeding, in particular to a rapid propagation technology of apple tissue culture seedlings.
Background
The apple seedling breeding technology comprises cuttage, layering, grafting, tissue culture and the like. The apple branch cuttage has low rooting rate and low survival rate, and a large number of plants are difficult to obtain in a short time; the propagation coefficient of the layering is low, the cost is high, and the technical requirement difficulty is high; the apple virus disease is easy to spread rapidly due to grafting, so that the quality of seedlings is influenced, the garden building quality is influenced, and the later-stage management cost is influenced; the tissue culture can rapidly obtain a large number of plants in a short time, the cost is low, the benefit is high, and the cost of the nursery stock can be greatly improved by breeding the virus-free tissue culture seedling.
At present, apple tissue seedling rapid propagation technologies are uneven, and formulas for propagation culture and rooting culture are full of liman, but the repeatability is poor and the application effect is poor; the survival rate of transplanted tissue culture seedlings is low, the cost is high, the commercialization progress is slow, and the popularization and the use of apple virus-free seedlings are restricted.
In summary, the problems of the prior art are as follows: the apple tissue culture seedling rapid propagation technology has poor repeatability, poor application effect and low transplanting survival rate.
Disclosure of Invention
The invention aims to solve the defects in the prior art and provides a rapid propagation method of apple tissue culture seedlings.
A rapid propagation method of apple tissue culture seedlings is characterized by comprising the following steps: culturing the apple tissue culture seedlings by using a subculture multiplication culture medium for 30-40 days, then transferring the apple tissue culture seedlings to a rooting culture medium for culturing for 25-35 days, and finally transplanting the rooted tissue culture seedlings to a transplanting substrate by using a nutrition pot;
the subculture multiplication medium is used for subculture multiplication of the apple tissue culture seedlings, the rooting medium is used for rooting culture of the tissue culture seedlings, and the transplanting medium is used for transplanting when the rooting of the tissue culture seedlings meets the transplanting requirement.
Further, in the rapid propagation method of tissue culture seedling of apple as described above, the subculture multiplication medium is calculated by using 1LMS medium as a minimal medium, and includes: 1900mg of potassium nitrate, 1650mg of ammonium nitrate, 370mg of magnesium sulfate heptahydrate, 170mg of potassium dihydrogen phosphate, 440mg of calcium chloride dihydrate, 22.3mg of manganese sulfate tetrahydrate, 8.6mg of zinc sulfate heptahydrate, 6.2mg of boric acid, 0.83mg of potassium iodide, 0.25mg of sodium molybdate dihydrate, 0.025mg of copper sulfate pentahydrate, 0.025mg of cobalt chloride hexahydrate, 27.8mg of ferric sulfate heptahydrate, 37.3mg of disodium ethylenediaminetetraacetate dihydrate, 100mg of inositol, 2mg of glycine, 0.5mg of nicotinic acid, 0.5mg of vitamin B60.5mg, vitamin B10.1mg, 6-BA0.8mg, IBA0.2mg, sucrose 40g, agar 6g, polyvinyl alcohol 2g, pH 6.0.
Further, in the method for rapid propagation of tissue culture seedling of apple as described above, the rooting medium takes 1L1/2MS medium as a minimal medium, and comprises: 950mg of potassium nitrate, 825mg of ammonium nitrate, 185mg of magnesium sulfate heptahydrate, 85mg of potassium dihydrogen phosphate, 220mg of calcium chloride dihydrate, 11.15mg of manganese sulfate tetrahydrate, 4.3mg of zinc sulfate heptahydrate, 3.1mg of boric acid, 0.415mg of potassium iodide, 0.125mg of sodium molybdate dihydrate, 0.0125mg of copper sulfate pentahydrate, 0.0125mg of cobalt chloride hexahydrate, 13.9mg of ferric sulfate heptahydrate, 18.65mg of disodium ethylenediaminetetraacetate dihydrate, 50mg of inositol, 1mg of glycine, 0.25mg of nicotinic acid, 0.25mg of vitamin B60.25mg, vitamin B10.05mg, IAA2.0mg, sucrose 20g, agar 6g, pH 6.0.
Further, according to the rapid propagation method of the tissue culture apple seedlings, the transplanting substrate comprises: 100-200 parts of northeast black carbon turfy soil; 100-200 parts of vermiculite with the aperture of 3-6 mm.
Has the advantages that:
the proliferation culture medium is added with macroelements, microelements and other organic elements necessary for plant growth, and also added with a plant growth regulator cytokinin 6-BA for promoting bud germination and regeneration and IBA for promoting cell differentiation and division, and the value increase coefficient of the tissue culture seedling can be obviously improved by adjusting the concentration of the two plant growth regulators, wherein the highest proliferation coefficient is achieved when the concentration of 6-BA is 0.8mg/L and IBA is 0.2mg/L, the average proliferation coefficient is 6.1, and the value increase coefficient is 8.9% compared with the prior art; the rooting culture medium is added with essential elements and plant growth regulator IAA alone, the rooting effect is achieved by changing the distribution of nutrient substances in the tissue culture seedlings, the rooting rate of the tissue culture seedlings can be obviously improved when the concentration of the plant growth regulator IAA is 2.0mg/L, the rooting rate is up to 85 percent on average, and the rooting rate is improved by 6.25 percent compared with the prior art; the selection of the species and the proportion of the transplanting matrix influences the transplanting survival rate of the tissue culture seedlings to a great extent, the Chinese herbal medicine charcoal can provide nutrition required by the growth of the tissue culture seedlings, the vermiculite with the aperture of 3-6mm provides gaps, the oxygen content is increased, the proportion of the two matrixes is 1:1, the survival rate of the tissue culture seedlings can be increased to over 75 percent, the highest survival rate can reach 85 percent, and the survival rate is increased by 6.25-7.14 percent compared with the prior art.
Drawings
FIG. 1 is a diagram showing the proliferation status of tissue culture seedlings provided in the examples of the present invention (the right diagram shows the formulation of the present application, and the left diagram shows the conventional formulation);
FIG. 2 is a diagram illustrating the rooting of tissue culture seedlings according to an embodiment of the present invention;
FIG. 3 is a first diagram illustrating the transplanting of tissue culture seedlings according to an embodiment of the present invention;
FIG. 4 is a second diagram illustrating the transplanting of tissue culture seedlings according to the embodiment of the present invention;
fig. 5 is a third diagram of the transplanting situation of the tissue culture seedlings provided by the embodiment of the invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the technical solutions of the present invention are described clearly and completely below, and it is obvious that the described embodiments are some, not all embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example (b):
firstly, preparing a subculture multiplication medium, a rooting medium culture medium and a transplanting medium; the subculture multiplication medium is used for subculture multiplication of the apple tissue culture seedlings, the rooting medium is used for rooting culture of the tissue culture seedlings, and the transplanting medium is used for transplanting when the rooting of the tissue culture seedlings meets the transplanting requirement.
Subculture multiplication medium:
adding about 13L of distilled water into a filling machine, adding 40g of polyvinyl alcohol, sequentially adding 120g of agar, 800g of cane sugar, 38g of potassium nitrate, 33g of ammonium nitrate, 7.4g of magnesium sulfate heptahydrate, 3.4g of potassium dihydrogen phosphate, 8.8g of calcium chloride dihydrate, 0.446g of manganese sulfate tetrahydrate, 0.172g of zinc sulfate heptahydrate, 0.124g of boric acid, 0.0166g of potassium iodide, 0.005g of sodium molybdate dihydrate, 0.0005g of copper sulfate pentahydrate, 0.0005g of cobalt chloride hexahydrate, 0.556g of ferric sulfate heptahydrate, 0.746g of disodium ethylenediaminetetraacetate dihydrate, 2g of inositol, 0.04g of glycine, 0.01g of nicotinic acid, 0.01g of vitamin B60.01g, vitamin B10.002g, 6-BA0.016g and NAA0.001g. Stopping heating after the water is boiled, and adding distilled water to a constant volume of 20L. The pH was adjusted to 6.0. And (5) after filling, sterilizing at high temperature and high pressure for 25 min.
Rooting culture medium:
adding about 13L of distilled water into a filling machine, and adding 120g of agar, 400g of cane sugar, 19g of potassium nitrate, 16.5g of ammonium nitrate, 3.7g of magnesium sulfate heptahydrate, 1.7g of monopotassium phosphate, 4.4g of calcium chloride dihydrate, 0.223g of manganese sulfate tetrahydrate, 0.086g of zinc sulfate heptahydrate, 0.062g of boric acid, 0.0083g of potassium iodide, 0.0025g of sodium molybdate dihydrate, 0.00025g of copper sulfate pentahydrate, 0.00025g of cobalt chloride hexahydrate, 0.278g of ferric sulfate heptahydrate, 0.373g of disodium ethylenediaminetetraacetate dihydrate, 1g of inositol, 0.02g of glycine, 0.005g of nicotinic acid and 0.005g of vitamin B into cold water in sequence60.005g, vitamin B10.001g and IAA0.04g. The heating is stopped after the water is boiled,adding distilled water to constant volume of 20L. The pH was adjusted to 6.0. And (5) after filling, sterilizing at high temperature and high pressure for 25 min.
Transplanting a substrate:
the transplanting substrate comprises: 150 parts of northeast black carbon turfy soil; 200 parts of vermiculite with the aperture of 3-6 mm.
Then culturing the apple tissue culture seedlings by using a subculture multiplication culture medium for 30 days, then transferring the apple tissue culture seedlings to a rooting culture medium for culturing for 30 days, and finally transplanting the rooted tissue culture seedlings to a transplanting substrate by using a nutrition pot. FIG. 1 is a diagram showing the proliferation of tissue culture seedlings according to an embodiment of the present invention; FIG. 2 is a diagram illustrating the rooting of tissue culture seedlings according to an embodiment of the present invention; FIG. 3 is a first diagram illustrating the transplanting of tissue culture seedlings according to an embodiment of the present invention; FIG. 4 is a second diagram illustrating the transplanting of tissue culture seedlings according to the embodiment of the present invention; fig. 5 is a third diagram of the transplanting situation of the tissue culture seedlings provided by the embodiment of the invention. As can be seen from FIGS. 1 to 5, the proliferation coefficient of the proliferation medium provided by the invention is up to 6.1 on average, which is 8.9% higher than that of the prior art; the rooting culture medium provided by the invention achieves the effect of rooting, remarkably improves the rooting rate of the tissue culture seedling, achieves 85 percent on average and is improved by 6.25 percent compared with the prior art; the survival rate of the tissue culture seedlings is improved by the transplanting matrix provided by the invention to over 75 percent and up to 85 percent, and is improved by 6.25 to 7.14 percent compared with the prior art.
Finally, it should be noted that: the above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.
Claims (4)
1. A rapid propagation method of apple tissue culture seedlings is characterized by comprising the following steps: culturing the apple tissue culture seedlings by using a subculture multiplication culture medium for 30-40 days, then transferring the apple tissue culture seedlings to a rooting culture medium for culturing for 25-35 days, and finally transplanting the rooted tissue culture seedlings to a transplanting substrate by using a nutrition pot;
the subculture multiplication medium is used for subculture multiplication of the apple tissue culture seedlings, the rooting medium is used for rooting culture of the tissue culture seedlings, and the transplanting medium is used for transplanting when the rooting of the tissue culture seedlings meets the transplanting requirement.
2. The rapid propagation method of tissue culture seedling of apple according to claim 1, wherein the subculture multiplication medium is calculated by taking 1L MS medium as a minimal medium, and comprises: 1900mg of potassium nitrate, 1650mg of ammonium nitrate, 370mg of magnesium sulfate heptahydrate, 170mg of potassium dihydrogen phosphate, 440mg of calcium chloride dihydrate, 22.3mg of manganese sulfate tetrahydrate, 8.6mg of zinc sulfate heptahydrate, 6.2mg of boric acid, 0.83mg of potassium iodide, 0.25mg of sodium molybdate dihydrate, 0.025mg of copper sulfate pentahydrate, 0.025mg of cobalt chloride hexahydrate, 27.8mg of ferric sulfate heptahydrate, 37.3mg of disodium ethylenediaminetetraacetate dihydrate, 100mg of inositol, 2mg of glycine, 0.5mg of nicotinic acid, 0.5mg of vitamin B60.5mg, vitamin B10.1mg, 6-BA0.8mg, IBA0.2mg, sucrose 40g, agar 6g, polyvinyl alcohol 2g, pH 6.0.
3. The method for rapid propagation of tissue culture seedling of apple according to claim 1, wherein the rooting medium takes 1L1/2MS medium as a minimal medium, and comprises: 950mg of potassium nitrate, 825mg of ammonium nitrate, 185mg of magnesium sulfate heptahydrate, 85mg of potassium dihydrogen phosphate, 220mg of calcium chloride dihydrate, 11.15mg of manganese sulfate tetrahydrate, 4.3mg of zinc sulfate heptahydrate, 3.1mg of boric acid, 0.415mg of potassium iodide, 0.125mg of sodium molybdate dihydrate, 0.0125mg of copper sulfate pentahydrate, 0.0125mg of cobalt chloride hexahydrate, 13.9mg of ferric sulfate heptahydrate, 18.65mg of disodium ethylenediaminetetraacetate dihydrate, 50mg of inositol, 1mg of glycine, 0.25mg of nicotinic acid, 0.25mg of vitamin B60.25mg, vitamin B10.05mg, IAA2.0mg, sucrose 20g, agar 6g, pH 6.0.
4. The rapid propagation method of tissue culture seedlings of apples according to claim 1, wherein the transplanting substrate comprises: 100-200 parts of northeast black carbon turfy soil; 100-200 parts of vermiculite with the aperture of 3-6 mm.
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| CN202010977539.8A CN111903529A (en) | 2020-09-17 | 2020-09-17 | Rapid propagation technology for apple tissue culture seedlings |
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| CN202010977539.8A CN111903529A (en) | 2020-09-17 | 2020-09-17 | Rapid propagation technology for apple tissue culture seedlings |
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106613934A (en) * | 2015-10-28 | 2017-05-10 | 北京市农林科学院 | Method for rapidly propagating apple rootstock SH6 |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106613934A (en) * | 2015-10-28 | 2017-05-10 | 北京市农林科学院 | Method for rapidly propagating apple rootstock SH6 |
Non-Patent Citations (2)
| Title |
|---|
| 吴瑞刚等: "苹果砧木‘青砧一号’离体高效再生体系的建立", 《植物生理学报》 * |
| 师校欣等: "苹果砧木离体培养中玻璃化问题的研究", 《河北农业大学学报》 * |
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