CN113661921A - Cultivation method of chionanthus roseus based on ethidium bromide and ultraviolet irradiation mutagenesis - Google Patents
Cultivation method of chionanthus roseus based on ethidium bromide and ultraviolet irradiation mutagenesis Download PDFInfo
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Abstract
The invention discloses a cultivation method of chionanthus roseus based on ethidium bromide and ultraviolet irradiation mutagenesis, which comprises the following steps: the method comprises the following steps of (1) sowing tassel seeds collected from the bottom of 10 months to the beginning of 11 months in a seedling tray after sprouting through sand storage, and managing according to a conventional method after sprouting; in the last ten days of 4 months, when the tassel seedlings come out of the earth, the cotyledons are flattened and the true leaves sprout, uniformly spraying a watering can filled with ethidium bromide solution on the stem tips and the surfaces of the true leaves of the tassel, spraying once in the morning, in the middle and at the evening each day, covering a plastic film and a sunshade net after spraying, irradiating the tassel with ultraviolet lamps for 3-6 days continuously, removing the ethidium bromide solution on the stem tips and the surfaces of the true leaves with clear water, and performing conventional management; one month later, the variation of the tassel leaves gradually appears, the variation is stable until 9 months, the formed variation plants show that the color of the leaves is changed into golden yellow, and the plants with the variation characters are placed in a greenhouse for conventional management. The method has high mutation rate of tassel, and high survival rate of tassel seedlings after mutation.
Description
Technical Field
The invention relates to the technical field of agriculture, in particular to a cultivation method of tassel trees with colored leaves based on ethidium bromide and ultraviolet irradiation mutagenesis.
Background
Chionanthus retusus, genus Chionanthus retusus of Oleaceae, deciduous shrub or arbor. The male and female heterozygotic plants, which are clear and pleasant, can be planted in gardens, can also be potted to make a stake scene for grafting sweet osmanthus, and are excellent local ornamental tree species. The chionanthus retusus has the characteristics of drought resistance, barren property, salt and alkali resistance, long service life and the like, can be used for ornamental, material and barren mountain afforestation, medicinal, tea frying, oil pressing and the like, and has wide application and high economic value. Under the influence of the factors of slow growth speed, low natural reproduction rate, artificial digging and the like, the chionanthus retusus is in a gradually dangerous state, and the fine variety breeding of specific germplasm resources is urgently needed. The systematic research on the flow threo tree species resources has not been carried out in China so far, the conventional breeding stage with lower utilization level is still realized, and the artificial induced mutant is one of the important ways for obtaining new plant germplasm. However, in the prior art, the research on artificially inducing the mutant to obtain the specific germplasm resources of tassel is less, so how to research the method for obtaining the specific germplasm resources of tassel by using the artificially induced mutant has important significance.
Disclosure of Invention
The technical problem to be solved by the invention is as follows: aiming at the defects in the prior art, the cultivation method of the tassel tree with colored leaves based on ethidium bromide and ultraviolet irradiation mutagenesis is provided, the method is high in tassel variation rate, and the survival rate of the mutated tassel seedlings is high.
In order to solve the technical problems, the technical scheme of the invention is as follows:
a cultivation method of chionanthus roseus based on ethidium bromide and ultraviolet irradiation mutagenesis comprises the following steps:
(1) the method comprises the following steps of (1) sowing tassel seeds collected from the bottom of 10 months to the beginning of 11 months in a seedling tray after sprouting through sand storage, and managing according to a conventional method after sprouting;
(2)4, in the last ten days of the month, when the seedlings of tassella virginiana come out, the cotyledons are flattened and the true leaves germinate, namely when the cotyledons are flattened and the true leaves emerge, a watering can filled with ethidium bromide solution with the concentration of 0.9-1.2mg/L is uniformly sprayed on the stem tips and the surfaces of the true leaves of the tassella virginiana, the tassels are sprayed once in the morning, in the middle and at the evening each day, a plastic film and a sun-shading net are covered after the spraying, ultraviolet lamp irradiation is carried out on the sunflowers, the treatment is carried out for 3-6 days continuously, then the ethidium bromide solution on the stem tips and the surfaces of the true leaves is removed by clear water, and the conventional management is carried out;
(3) one month later, the leaves of the tassel gradually change, the color of the leaves begins to turn yellow, the change is stable from 9 months, the formed variant plants show that the color of the leaves changes to golden yellow, and the plants with the variant characters are placed in a greenhouse for conventional management.
Preferably, when the seeds after sand storage and sprouting are sowed in the seedling raising tray, the matrix in the seedling raising tray comprises 20-30 parts by weight of mushroom residues, 20-30 parts by weight of turf, 20-30 parts by weight of coconut shell powder and 10-20 parts by weight of functional filler.
Preferably, the functional filler is a multilayer filler which takes montmorillonite/lignin composite material and chitosan cross-linked coated urea as a core layer and takes super absorbent polymer as a shell layer.
Preferably, the preparation method of the functional filler comprises the following steps:
1) adding organic solvent lignin into a sodium hydroxide solution, stirring and mixing for 30-50min, adding 2, 3-epoxypropyl trimethyl ammonium chloride, continuously stirring for 3-5h, and finally performing vacuum drying to obtain quaternized lignin;
2) preparing a urea solution with the concentration of 0.5-1.5g/ml, heating to 75-85 ℃, adding a sodium alginate solution with the concentration of 2 wt% and a mixed solution of montmorillonite and quaternized lignin into the urea solution, wherein the concentrations of the montmorillonite and the quaternized lignin in the mixed solution are both 2 wt%, stirring to form gel, slowly dripping the gel into a calcium chloride solution with the concentration of 4 wt% by using an injector, stirring for 20-50min, filtering, and drying the solid to prepare a core layer material;
3) and coating a polyacrylamide solution on the surface of the core layer material, and drying to obtain the multifunctional filler.
Preferably, in the step 1), the concentration of the sodium hydroxide solution is 15-20 wt%, and the mass ratio of the organic solvent type lignin, the sodium hydroxide solution and the 2, 3-epoxypropyltrimethylammonium chloride is (2-3): 30: (0.4-0.6).
Preferably, in the step 2), the volume ratio of the urea solution, the sodium alginate solution, the mixed solution of montmorillonite and quaternized lignin is 10 (30-31) to (15-20).
Preferably, in step 3), the concentration of the polyacrylamide solution is 10mg/ml, the molecular weight of the polyacrylamide is 500 to 2000 ten thousand, and the ratio of the core layer material to the polyacrylamide solution is 1 g: (20-30) ml.
Preferably, in the step (2), the concentration of the ethidium bromide solution is 1 mg/L; the power of the ultraviolet lamp is 30W; the duration of the continuous treatment was 5 days.
As an improvement of the technical scheme, in the step (3), when the plants with the mutation characters are placed in a greenhouse for conventional management, the mixed nutrient solution of potassium dihydrogen phosphate and urea is sprayed on the plants once every 15 days.
Preferably, the mixed nutrient solution has a mass concentration of monopotassium phosphate and a mass concentration of urea of 0.2% and 0.5%, respectively.
Due to the adoption of the technical scheme, the invention has the beneficial effects that:
ethidium bromide is a strong mutagen and can be embedded into basic group molecules to cause mismatching, inhibit cell growth or initiate site-directed mutagenesis.
During cultivation, the self-made functional filler is added into the matrix of the seedling tray, the functional filler adopts montmorillonite/lignin nano hybrid and alginate as an encapsulating layer to encapsulate urea, and then a super absorbent polymer is adopted to carry out outer layer encapsulation to form the multi-layer encapsulated filler, so that the multi-layer encapsulated filler has strong water retention capacity and certain slow release performance, can effectively provide nutrition required during the growth period of the tassel seedlings and promotes the growth of the tassel seedlings. During the preparation. The prepared mixed solution of the quaternized lignin, the montmorillonite and the sodium alginate is mixed with the urea solution, the network skeleton of the quaternized lignin can effectively enter the montmorillonite to form a lignin/montmorillonite nano hybrid, the prepared gel is dripped into the calcium chloride solution, the alginate forms a network polymer under the action of the calcium chloride, the lignin/montmorillonite nano hybrid and the urea are wrapped inside, and finally the surface of the gel is wrapped with the high water absorption polymer, so that the prepared functional filler has excellent slow release performance and water retention performance.
The planting method of the tassel seedlings, provided by the invention, is simple to operate, and the variation rate and survival rate of the plants are high after the plants are induced by the ethidium bromide solution.
Detailed Description
The invention is further illustrated by the following examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention.
In the following examples and comparative examples, the number of treated plants was 300 in the mutation treatment.
Example 1
(1) Adding 20g of organic solvent type lignin into 300g of 15 wt% sodium hydroxide solution, stirring and mixing for 30min, then adding 4g of 2, 3-epoxypropyltrimethylammonium chloride, continuing stirring for 3h, and finally performing vacuum drying to obtain quaternized lignin;
(2) preparing a urea solution with the concentration of 0.5g/ml, heating to 75 ℃, adding 300ml of a sodium alginate solution with the concentration of 2 wt% and 150ml of a mixed solution of montmorillonite and quaternized lignin into 100ml of the urea solution, wherein the concentrations of the montmorillonite and the quaternized lignin in the mixed solution are both 2 wt%, stirring to form gel, slowly dripping the gel into 500ml of a calcium chloride solution with the concentration of 4 wt% by using an injector, stirring for 20min, filtering, and drying the solid to prepare a core layer material;
(3) coating 300ml of polyacrylamide solution with the concentration of 10mg/ml on the surface of 10g of the core layer material, and drying to obtain the multifunctional filler; mixing 20 parts of mushroom residues, 20 parts of turf, 20 parts of coconut shell powder and 10 parts of functional filler in parts by weight to prepare a matrix;
(4) the method comprises the following steps of (1) sowing tassel seeds collected from the bottom of 10 months to the beginning of 11 months in a seedling tray filled with the substrate after sand storage and sprouting, and managing according to a conventional method after sprouting; when the seedlings of tassel come out of the earth in the last ten days of 4 months, flattening cotyledons and exposing true leaves, uniformly spraying a watering can filled with ethidium bromide solution with the concentration of 0.9mg/L on the tips of the stems and the surfaces of the true leaves, spraying once in the morning, in the middle and at night every day, covering a plastic film and a sunshade net after spraying, irradiating the stems and the true leaves by using an ultraviolet lamp with the power of 30W, continuously treating for 5 days, removing the ethidium bromide solution on the tips of the stems and the surfaces of the true leaves by using clean water, and performing conventional management; after one month, gradually changing the color of the tassel leaves, starting to turn yellow, stably changing the color of the leaves in 9 months, enabling the formed variant plants to show that the color of the leaves is changed into golden yellow, putting the plants with variant characters in a greenhouse for conventional management, spraying mixed nutrient solution of potassium dihydrogen phosphate and urea to the plants, and spraying once every 15 days; in the mixed nutrient solution, the mass concentrations of the monopotassium phosphate and the urea are respectively 0.2 percent and 0.5 percent.
Example 2
(1) Adding 30g of organic solvent type lignin into 300g of 20 wt% sodium hydroxide solution, stirring and mixing for 50min, then adding 6g of 2, 3-epoxypropyltrimethylammonium chloride, continuing stirring for 3-5h, and finally performing vacuum drying to obtain quaternized lignin;
(2) preparing 1.5g/ml urea solution, heating to 85 ℃, adding 300ml of 2 wt% sodium alginate solution and 150ml of mixed solution of montmorillonite and quaternized lignin into 100ml of urea solution, stirring until gel is formed, slowly dripping the gel into 500ml of 4 wt% calcium chloride solution by using an injector, stirring for 50min, filtering, and drying the solid to obtain a core layer material;
(3) coating 300ml of polyacrylamide solution with the concentration of 10mg/ml on the surface of 10g of the core layer material, and drying to obtain the multifunctional filler; mixing 30 parts of mushroom residues, 30 parts of turf, 30 parts of coconut shell powder and 20 parts of functional filler in parts by weight to prepare a matrix;
(4) the method comprises the following steps of (1) sowing tassel seeds collected from the bottom of 10 months to the beginning of 11 months in a seedling tray filled with the substrate after sand storage and sprouting, and managing according to a conventional method after sprouting; when the seedlings of tassel come out of the earth in the last ten days of 4 months, flattening cotyledons and exposing true leaves, uniformly spraying a watering can filled with ethidium bromide solution with the concentration of 1.0mg/L on the tips of the stems and the surfaces of the true leaves, spraying once in the morning, in the middle and at night every day, covering a plastic film and a sunshade net after spraying, irradiating the stems and the true leaves by using an ultraviolet lamp with the power of 30W, continuously treating for 5 days, removing the ethidium bromide solution on the tips of the stems and the surfaces of the true leaves by using clean water, and performing conventional management; after one month, gradually changing the color of the tassel leaves, starting to turn yellow, stably changing the color of the leaves in 9 months, enabling the formed variant plants to show that the color of the leaves is changed into golden yellow, putting the plants with variant characters in a greenhouse for conventional management, spraying mixed nutrient solution of potassium dihydrogen phosphate and urea to the plants, and spraying once every 15 days; in the mixed nutrient solution, the mass concentrations of the monopotassium phosphate and the urea are respectively 0.2 percent and 0.5 percent.
Example 3
(1) Adding 25g of organic solvent type lignin into 300g of 15 wt% sodium hydroxide solution, stirring and mixing for 40min, then adding 4.5g of 2, 3-epoxypropyltrimethylammonium chloride, continuing stirring for 3h, and finally performing vacuum drying to obtain quaternized lignin;
(2) preparing a urea solution with the concentration of 1g/ml, heating to 80 ℃, adding 300ml of a sodium alginate solution with the concentration of 2 wt% and 150ml of a mixed solution of montmorillonite and quaternized lignin into 100ml of the urea solution, wherein the concentrations of the montmorillonite and the quaternized lignin in the mixed solution are both 2 wt%, stirring to form gel, slowly dripping the gel into 500ml of a calcium chloride solution with the concentration of 4 wt% by using an injector, stirring for 30min, filtering, and drying the solid to prepare a core layer material;
(3) coating 300ml of polyacrylamide solution with the concentration of 10mg/ml on the surface of 10g of the core layer material, and drying to obtain the multifunctional filler; mixing 20 parts of mushroom residues, 30 parts of turf, 20 parts of coconut shell powder and 20 parts of functional filler in parts by weight to prepare a matrix;
(4) the method comprises the following steps of (1) sowing tassel seeds collected from the bottom of 10 months to the beginning of 11 months in a seedling tray filled with the substrate after sand storage and sprouting, and managing according to a conventional method after sprouting; when the seedlings of tassel come out of the earth in the last ten days of 4 months, flattening cotyledons and exposing true leaves, uniformly spraying a watering can filled with ethidium bromide solution with the concentration of 1.1mg/L on the tips of the stems and the surfaces of the true leaves, spraying once in the morning, in the middle and at night every day, covering a plastic film and a sunshade net after spraying, irradiating the stems and the true leaves by using an ultraviolet lamp with the power of 30W, continuously treating for 5 days, removing the ethidium bromide solution on the tips of the stems and the surfaces of the true leaves by using clean water, and performing conventional management; after one month, gradually changing the color of the tassel leaves, starting to turn yellow, stably changing the color of the leaves in 9 months, enabling the formed variant plants to show that the color of the leaves is changed into golden yellow, putting the plants with variant characters in a greenhouse for conventional management, spraying mixed nutrient solution of potassium dihydrogen phosphate and urea to the plants, and spraying once every 15 days; in the mixed nutrient solution, the mass concentrations of the monopotassium phosphate and the urea are respectively 0.2 percent and 0.5 percent.
Example 4
(1) Adding 25g of organic solvent type lignin into 300g of 20 wt% sodium hydroxide solution, stirring and mixing for 30min, then adding 5g of 2, 3-epoxypropyltrimethylammonium chloride, continuing stirring for 3h, and finally performing vacuum drying to obtain quaternized lignin;
(2) preparing a urea solution with the concentration of 1g/ml, heating to 80 ℃, adding 300ml of a sodium alginate solution with the concentration of 2 wt% and 150ml of a mixed solution of montmorillonite and quaternized lignin into 100ml of the urea solution, wherein the concentrations of the montmorillonite and the quaternized lignin in the mixed solution are both 2 wt%, stirring to form gel, slowly dripping the gel into 500ml of a calcium chloride solution with the concentration of 4 wt% by using an injector, stirring for 30min, filtering, and drying the solid to prepare a core layer material;
(3) coating 300ml of polyacrylamide solution with the concentration of 10mg/ml on the surface of 10g of the core layer material, and drying to obtain the multifunctional filler; mixing 25 parts of mushroom residues, 25 parts of turf, 30 parts of coconut shell powder and 15 parts of functional filler by weight to prepare a matrix;
(4) the method comprises the following steps of (1) sowing tassel seeds collected from the bottom of 10 months to the beginning of 11 months in a seedling tray filled with the substrate after sand storage and sprouting, and managing according to a conventional method after sprouting; when the seedlings of tassel come out of the earth in the last ten days of 4 months, flattening cotyledons and exposing true leaves, uniformly spraying a watering can filled with ethidium bromide solution with the concentration of 1.2mg/L on the tips of the stems and the surfaces of the true leaves, spraying once in the morning, in the middle and at night every day, covering a plastic film and a sunshade net after spraying, irradiating the stems and the true leaves by using an ultraviolet lamp with the power of 30W, continuously treating for 5 days, removing the ethidium bromide solution on the tips of the stems and the surfaces of the true leaves by using clean water, and performing conventional management; after one month, gradually changing the color of the tassel leaves, starting to turn yellow, stably changing the color of the leaves in 9 months, enabling the formed variant plants to show that the color of the leaves is changed into golden yellow, putting the plants with variant characters in a greenhouse for conventional management, spraying mixed nutrient solution of potassium dihydrogen phosphate and urea to the plants, and spraying once every 15 days; in the mixed nutrient solution, the mass concentrations of the monopotassium phosphate and the urea are respectively 0.2 percent and 0.5 percent.
Example 5
(1) Adding 30g of organic solvent type lignin into 300g of 15 wt% sodium hydroxide solution, stirring and mixing for 40min, then adding 5.5g of 2, 3-epoxypropyl trimethyl ammonium chloride, continuing stirring for 3h, and finally performing vacuum drying to obtain quaternized lignin;
(2) preparing a urea solution with the concentration of 1g/ml, heating to 80 ℃, adding 300ml of a sodium alginate solution with the concentration of 2 wt% and 150ml of a mixed solution of montmorillonite and quaternized lignin into 100ml of the urea solution, wherein the concentrations of the montmorillonite and the quaternized lignin in the mixed solution are both 2 wt%, stirring to form gel, slowly dripping the gel into 500ml of a calcium chloride solution with the concentration of 4 wt% by using an injector, stirring for 30min, filtering, and drying the solid to prepare a core layer material;
(3) coating 250ml of polyacrylamide solution with the concentration of 10mg/ml on the surface of 10g of the core layer material, and drying to obtain the multifunctional filler; mixing 25 parts of mushroom residues, 25 parts of turf, 25 parts of coconut shell powder and 10 parts of functional filler by weight to prepare a matrix;
(4) the method comprises the following steps of (1) sowing tassel seeds collected from the bottom of 10 months to the beginning of 11 months in a seedling tray filled with the substrate after sand storage and sprouting, and managing according to a conventional method after sprouting; when the seedlings of tassel come out of the earth in the last ten days of 4 months, flattening cotyledons and exposing true leaves, uniformly spraying a watering can filled with ethidium bromide solution with the concentration of 1.0mg/L on the tips of the stems and the surfaces of the true leaves, spraying once in the morning, in the middle and at night every day, covering a plastic film and a sunshade net after spraying, irradiating the stems and the true leaves by using an ultraviolet lamp with the power of 30W, continuously treating for 5 days, removing the ethidium bromide solution on the tips of the stems and the surfaces of the true leaves by using clean water, and performing conventional management; after one month, gradually changing the color of the tassel leaves, starting to turn yellow, stably changing the color of the leaves in 9 months, enabling the formed variant plants to show that the color of the leaves is changed into golden yellow, putting the plants with variant characters in a greenhouse for conventional management, spraying mixed nutrient solution of potassium dihydrogen phosphate and urea to the plants, and spraying once every 15 days; in the mixed nutrient solution, the mass concentrations of the monopotassium phosphate and the urea are respectively 0.2 percent and 0.5 percent.
Comparative example 1
Water was used instead of the ethidium bromide solution, and the other conditions were the same as in example 5.
Comparative example 2
The concentration of the ethidium bromide solution was 0.5mg/L, and the other conditions were the same as in example 5.
Comparative example 3
The concentration of the ethidium bromide solution was 0.6mg/L, and the other conditions were the same as in example 5.
Comparative example 4
The concentration of the ethidium bromide solution was 0.7mg/L, and the other conditions were the same as in example 5.
Comparative example 5
The concentration of the ethidium bromide solution was 0.8mg/L, and the other conditions were the same as in example 5.
Comparative example 6
The concentration of the ethidium bromide solution was 1.3mg/L, and the other conditions were the same as in example 5.
Comparative example 7
The concentration of the ethidium bromide solution was 1.4mg/L, and the other conditions were the same as in example 5.
Comparative example 8
Coconut husk powder is used in the matrix to replace the functional filler, and other conditions are the same as in example 5.
The growth states of the tassel seedlings cultured in the above examples and comparative examples were followed, and the conditions of the plants were counted when they were to be transplanted, with the results shown in table 1.
TABLE 1
From the above results it can be seen that: the fringe seedlings treated by the ethidium bromide solution have variation, the variation rate of plants is improved along with the improvement of the concentration of the ethidium bromide solution, but when the concentration of the ethidium bromide solution is too high, the variation rate of the plants is reduced, the death rate is increased, and therefore when the concentration of the ethidium bromide solution is controlled to be 0.9-1.2mg/L, the variation rate and the survival rate of the plants are higher. In addition, the self-made functional filler is added into the matrix of the seedling-raising tray, so that the survival rate of the tassel seedlings is improved to a certain extent.
Further, it should be understood that various changes or modifications of the present invention may be made by those skilled in the art after reading the teaching of the present invention, and such equivalents may fall within the scope of the present invention as defined in the appended claims.
Claims (10)
1. A cultivation method of chionanthus roseus based on ethidium bromide and ultraviolet irradiation mutagenesis is characterized by comprising the following steps:
(1) the method comprises the following steps of (1) sowing tassel seeds collected from the bottom of 10 months to the beginning of 11 months in a seedling tray after sprouting through sand storage, and managing according to a conventional method after sprouting;
(2) when the seedlings of tassel come out of the earth in the last ten days of 4 months, the cotyledons are flattened and the true leaves sprout, uniformly spraying a watering can filled with ethidium bromide solution with the mass concentration of 0.9-1.2mg/L on the stem tips and the surfaces of the tassel and the true leaves once a day in the morning, in the middle and at night, covering a plastic film and a sunshade net after spraying, irradiating the tassel and the true leaves with ultraviolet lamps for 3-6 days, and then removing the ethidium bromide solution on the stem tips and the surfaces of the true leaves with clear water for conventional management;
(3) one month later, the leaves of the tassel gradually change, the color of the leaves begins to turn yellow, the change is stable from 9 months, the formed variant plants show that the color of the leaves changes to golden yellow, and the plants with the variant characters are placed in a greenhouse for conventional management.
2. The cultivation method of chionanthus roseus based on ethidium bromide and ultraviolet irradiation mutagenesis as claimed in claim 1, wherein: when seeds after sand storage and sprouting are sowed in a seedling raising plate, the matrix in the seedling raising plate comprises 20-30 parts by weight of mushroom residues, 20-30 parts by weight of turf, 20-30 parts by weight of coconut shell powder and 10-20 parts by weight of functional filler.
3. The cultivation method of chionanthus roseus based on ethidium bromide and ultraviolet irradiation mutagenesis as claimed in claim 2, wherein: the functional filler is a multilayer filler which takes montmorillonite/lignin composite material and chitosan cross-linked coated urea as a core layer and takes super absorbent polymer as a shell layer.
4. The cultivation method of chionanthus roseus based on ethidium bromide and ultraviolet irradiation mutagenesis as claimed in claim 3, wherein: the preparation method of the functional filler comprises the following steps:
1) adding organic solvent lignin into a sodium hydroxide solution, stirring and mixing for 30-50min, adding 2, 3-epoxypropyl trimethyl ammonium chloride, continuously stirring for 3-5h, and finally performing vacuum drying to obtain quaternized lignin;
2) preparing a urea solution with the concentration of 0.5-1.5g/ml, heating to 75-85 ℃, adding a sodium alginate solution with the concentration of 2 wt% and a mixed solution of montmorillonite and quaternized lignin into the urea solution, wherein the concentrations of the montmorillonite and the quaternized lignin in the mixed solution are both 2 wt%, stirring to form gel, slowly dripping the gel into a calcium chloride solution with the concentration of 4 wt% by using an injector, stirring for 20-50min, filtering, and drying the solid to prepare a core layer material;
3) and coating a polyacrylamide solution on the surface of the core layer material, and drying to obtain the multifunctional filler.
5. The cultivation method of chionanthus roseus based on ethidium bromide and ultraviolet irradiation mutagenesis as claimed in claim 4, wherein: in the step 1), the concentration of the sodium hydroxide solution is 15-20 wt%, and the mass ratio of the organic solvent type lignin, the sodium hydroxide solution and the 2, 3-epoxypropyltrimethylammonium chloride is (2-3): 30: (0.4-0.6).
6. The cultivation method of chionanthus roseus based on ethidium bromide and ultraviolet irradiation mutagenesis as claimed in claim 4, wherein: in the step 2), the volume ratio of the urea solution, the sodium alginate solution, the mixed solution of the montmorillonite and the quaternized lignin is 10 (30-31) to (15-20).
7. The cultivation method of chionanthus roseus based on ethidium bromide and ultraviolet irradiation mutagenesis as claimed in claim 4, wherein: in the step 3), the concentration of the polyacrylamide solution is 10mg/ml, the molecular weight of the polyacrylamide is 500-2000 ten thousand, and the dosage ratio of the core layer material to the polyacrylamide solution is 1 g: (20-30) ml.
8. The cultivation method of chionanthus roseus based on ethidium bromide and ultraviolet irradiation mutagenesis as claimed in claim 1, wherein: in the step (2), the concentration of the ethidium bromide solution is 1 mg/L; the power of the ultraviolet lamp is 30W; the duration of the continuous treatment was 5 days.
9. The cultivation method of chionanthus roseus based on ethidium bromide and ultraviolet irradiation mutagenesis as claimed in claim 1, wherein: in the step (3), when plants with the variation character are placed in a greenhouse for conventional management, spraying mixed nutrient solution of potassium dihydrogen phosphate and urea to the plants once every 15 days.
10. The cultivation method of chionanthus roseus based on ethidium bromide and ultraviolet irradiation mutagenesis as claimed in claim 9, wherein: in the mixed nutrient solution, the mass concentrations of the monopotassium phosphate and the urea are respectively 0.2 percent and 0.5 percent.
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Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN106068757A (en) * | 2016-06-14 | 2016-11-09 | 泰安市泰山林业科学研究院 | A kind of method of mutagenesis of wingceltis coloured silk leaf kind |
CN109275562A (en) * | 2018-10-12 | 2019-01-29 | 泰安市泰山林业科学研究院 | A kind of radiaction mutation method of acer truncatum seed |
CN110036907A (en) * | 2019-03-22 | 2019-07-23 | 泰安市泰山林业科学研究院 | A kind of chemical mutagenesis combines the application in Chionanthus retusus polyploid breeding with negative pressure method |
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CN106068757A (en) * | 2016-06-14 | 2016-11-09 | 泰安市泰山林业科学研究院 | A kind of method of mutagenesis of wingceltis coloured silk leaf kind |
CN109275562A (en) * | 2018-10-12 | 2019-01-29 | 泰安市泰山林业科学研究院 | A kind of radiaction mutation method of acer truncatum seed |
CN110036907A (en) * | 2019-03-22 | 2019-07-23 | 泰安市泰山林业科学研究院 | A kind of chemical mutagenesis combines the application in Chionanthus retusus polyploid breeding with negative pressure method |
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