CN111955349A - Special antibacterial rooting culture medium for Zealand chrysanthemum, preparation method thereof and rooting culture method - Google Patents
Special antibacterial rooting culture medium for Zealand chrysanthemum, preparation method thereof and rooting culture method Download PDFInfo
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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Abstract
The invention relates to the technical field of biology, in particular to a special antibacterial rooting culture medium for Zea mays and a preparation method and a rooting culture method thereof, wherein the culture medium comprises the following raw materials in parts by weight: a major element; ferrous sulfate; ethylenediaminetetraacetic acid disodium salt; sodium mesh acid; manganese sulfate; zinc sulfate; copper sulfate; cobalt chloride; boric acid; a-naphthylacetic acid; indolebutyric acid; nano silver solution; penicillin; streptomycin and a coagulant. The invention has the advantages of simple formula, simple preparation method, low cost, no high-temperature disinfection and pollution resistance, short growth period in the seedling cultivation process, only 20-25 days, quick propagation, rooting in 5-7 days, 100% of rooting rate, 96-100% of seedling rate and 98-100% of transplanting survival rate in 30 days.
Description
Technical Field
The invention relates to the technical field of biology, in particular to an antibacterial rooting culture medium special for Zealand chrysanthemum, a preparation method thereof and a rooting culture method.
Background
Chrysanthemum is a plant of the Compositae, originally recorded in Shen nong Ben Cao Jing (Shen nong's herbal), listed as the superior product, and the traditional theory considers that chrysanthemum is sweet in nature, bitter in taste and slightly cold; entering lung and liver meridians; has effects in dispelling pathogenic wind, calming liver, improving eyesight, and clearing away heat and toxic materials. It is used for treating wind-heat type common cold, headache, vertigo, conjunctival congestion, swelling and pain, and blurred vision. The chrysanthemum can be used as a medicine for chrysanthemum in the market, while the chrysanthemum can be used as a medicine for chrysanthemum in the market for other medicinal chrysanthemum which is not recorded in pharmacopoeia.
In China, the quality and resources of the chrysanthemum are diverse, about 22 medicinal chrysanthemums are produced, the chrysanthemum is distributed all over the country, the cultivation history of the chrysanthemum is long, and the introduction behaviors are common. Zealand chrysanthemum is perennial herb perennial root plant, prefers warm and humid environment with sufficient sunlight, and is high quality variety bred by introduction, cultivation and breeding, belonging to ecological and economic perennial herb flower. The plant has both edible value and ornamental value, and both ornamental value and edible value; the tea can be drunk and eaten, can be drunk as tea, has the functions of health care and disease treatment, can be used for cooking vegetables and soup, and has homology of medicine and food; not only can be potted and arranged in families, but also can be used as landscaping plants for beautifying and greening the environment; can be used as economic plants in forest land, and can also be used as crops, gardening flowers and landscaping plants. The yield of the 'Zengcheng honey chrysanthemum' is high, the yield of a single plant is up to 100-. Moreover, the great-town honey chrysanthemum has the outstanding advantages of drought resistance and barren resistance, and experiments show that the cultivated great-town honey chrysanthemum does not compete with grains, vegetables and the like for cultivated land, can utilize the cultivated plants through ecological development of barren slopes, and can replace the cultivated land for other planting industries, thereby relieving the problem of outstanding contradiction between human lands in China.
However, researches show that after a common edible chrysanthemum variety is planted and cultivated for many years under natural conditions, the germplasm resources of the excellent variety can be quickly degraded, the excellent variety can not be well applied, and the quality can not be guaranteed. Therefore, tissue culture is required, and the tissue culture industrialized seedling culture mode is an important means for providing seedlings for modern chrysanthemum planting and cultivation. The intensive study on the culture medium of tissue culture makes the large-scale development of chrysanthemum planting possible. For example, chinese patent document CN107211896A discloses a culture medium and a method for open field chrysanthemum flap tissue culture, wherein the culture medium comprises an induction medium, a differentiation medium and a rooting medium; the induction culture medium is as follows: BA + NAA + sucrose +8g/L agar in MS culture medium; the differentiation culture medium is an MS culture medium of BA + NAA + sucrose + agar; the rooting culture medium is 1/2MS culture medium of NAA +3 sucrose + agar, and can improve the induction rate, differentiation rate and test-tube plantlet rooting rate of the open-field chrysanthemum callus; for example, chinese document CN104686366A discloses a tissue culture medium of wild chrysanthemum, which includes a subculture medium and a rooting medium, both of which include major elements such as potassium nitrate and ammonium nitrate, trace elements such as manganese sulfate monohydrate, zinc sulfate heptahydrate and boric acid, organic substances such as glycine, thiamine hydrochloride and pyridoxine hydrochloride, and plant growth regulators such as naphthylacetic acid, and the wild chrysanthemum has high rooting rate and transplanting survival rate.
However, these culture mediums are commonly MS culture mediums and are supplemented with a certain amount of different types of phytohormones, which are liable to cause pollution to tissue culture seedlings during the addition process, and the added components are more, resulting in an increase in the comprehensive cost, and therefore, there is a need to provide an antibacterial rooting culture medium special for coreopsis henryi, a preparation method thereof, and a rooting culture method thereof, which have the advantages of short growth cycle, rapid propagation, high transplanting survival rate, low pollution rate, and low comprehensive cost.
Disclosure of Invention
Aiming at the defects in the prior art, one of the purposes of the invention is to provide the antibacterial rooting medium special for the Zealand chrysanthemum and the preparation method thereof, the formula is simple, the preparation method is simple, the cost is low, high-temperature disinfection and pollution resistance are avoided, the growth cycle in the seedling cultivation process is short, only 20-25 days are needed, the propagation is rapid, the rooting can be realized in 5-7 days, the rooting rate is 100%, the seedling rate is 96-100%, and the transplanting survival rate in 30 days is 98-100%.
The above object of the present invention is achieved by the following technical solutions:
the antibacterial rooting culture medium special for the Zealand chrysanthemum comprises the following raw materials in parts by weight:
1800-2950 mg/L macroelements; 20-30 mg/L of ferrous sulfate; 30-40 mg/L of ethylene diamine tetraacetic acid disodium salt; 0.1-0.5 mg/L sodium mesh; 10-30 mg/L of manganese sulfate; 5-15 mg/L of zinc sulfate; 0.01-0.03 mg/L of copper sulfate; 0.01-0.03 mg/L of cobalt chloride; boric acid 4-8 mg/L; 0.05-0.2 mg/L of alpha-naphthylacetic acid; 0.1-1 mg/L of indolebutyric acid; 10-40 ml of nano silver solution; 10000-20000 units of penicillin; 8000-14000 units of streptomycin and 2-10 g/L of coagulant.
Preferably, the antibacterial rooting culture medium special for the Zealand chrysanthemum comprises the following raw materials in parts by weight:
2400-2950 mg/L macroelements; 25-30 mg/L of ferrous sulfate; 35-40 mg/L of ethylene diamine tetraacetic acid disodium salt; 0.2-0.5 mg/L of sodium mesh; 20-30 mg/L of manganese sulfate; 8-15 mg/L of zinc sulfate; 0.02-0.03 mg/L of copper sulfate; 0.02-0.03 mg/L of cobalt chloride; 5-8 mg/L of boric acid; 0.05-0.15 mg/L of alpha-naphthylacetic acid; 0.3-1 mg/L of indolebutyric acid; 10-30 ml of nano silver solution; 15000-20000 units of penicillin; 8000-12000 units of streptomycin and 4-10 g/L of coagulant.
Preferably, the weight content percentage of the nano silver solution is 1-4 wt%.
Preferably, the weight content percentage of the nano silver solution is 2 wt%.
Preferably, the macroelements include ammonia nitrate, potassium dihydrogen phosphate, calcium chloride and magnesium sulfate.
Preferably, the coagulating agent comprises carrageenan.
A preparation method of an antibacterial rooting culture medium special for Zealand chrysanthemum comprises the following steps:
s1, stirring and mixing the major elements, ferrous sulfate and ethylene diamine tetraacetic acid disodium salt according to the weight ratio to form a mixed solution A;
s2, stirring and mixing the nano-silver solution, the penicillin and the streptomycin in the weight ratio to form a mixed solution B;
s3, raising the temperature of the mixed solution A to 80-90 ℃, adding sodium mesh, manganese sulfate, zinc sulfate, copper sulfate, cobalt chloride, boric acid, alpha-naphthylacetic acid, indolebutyric acid and a coagulant in the weight ratio, stirring and mixing until carrageenan is completely dissolved to form a mixed solution C;
and S4, reducing the temperature of the mixed solution C to 60-70 ℃, adding the mixed solution B into the mixed solution C, uniformly stirring, and adjusting the pH value to 4.2-6.3 to obtain the antibacterial rooting culture medium special for the Zea mays.
Preferably, the pH value of the antibacterial rooting medium special for the Zealand chrysanthemum is 5.8.
The invention also provides a rooting culture method of the Zealand chrysanthemum, which comprises the following steps:
A. taking tender shoots in the excellent-shape subculture proliferation material of the Zealand chrysanthemum as an inoculation material to inoculate in the special antibacterial rooting culture medium for the Zealand chrysanthemum to obtain chrysanthemum tissue culture seedlings, binding the chrysanthemum tissue culture seedlings for 2-3 circles by using a food-grade preservative film along the joint of the bottle cap and the bottle, and keeping the bottle cap and the two sides of the bottle to be respectively wrapped by 1-1.5 cm;
B. after the tissue culture seedlings of the chrysanthemum are sealed by a preservative film and at the joint of a bottle and a bottle, the tissue culture seedlings are placed on a seedling hardening plastic greenhouse internal frame for rooting and seedling hardening culture at the temperature of 15-35 ℃, the illumination intensity of 8000-15000 Lux, the illumination time of 7-11 h and the humidity of 35-85% every day, the rooting and seedling hardening culture is carried out for 20-25 d, and the seedlings can be transplanted when the roots are tidy and the roots are 3-7 cm high.
According to the invention, organic matters and sucrose in the MS culture medium are removed innovatively, so that the culture medium is simplified, the cost of the culture medium is greatly reduced, the preparation process of the culture medium is simplified, and the risk of pollution caused by the organic matters and the sucrose is reduced; then adding bactericides such as nano-silver solution, penicillin, streptomycin and the like and antibiotics, so that the pollution rate of the culture medium can be effectively controlled without high-temperature sterilization, and the pollution rate is reduced to be below 3%;
in addition, the invention unexpectedly discovers that the growth cycle of the seedling can be shortened by compounding the nano-silver solution and the penicillin, compared with the growth cycle of 35-40 days of the conventional culture medium in the existing market, the growth cycle of the seedling is 20-25 days, the seedling can take roots in 5-7 days, the rooting rate can be increased to 100%, the seedling rate is 96-100%, and the transplanting survival rate in 30 days is 98-100%.
By sealing the preservative film and controlling environmental factors in the tissue culture process, the rooting and seedling hardening can be directly carried out in the natural environment of the plastic greenhouse, the pollution rate in the rooting and seedling hardening can be greatly reduced, and the seedling hardening time can be shortened. By combining the application of the culture medium, the growth period in the tissue culture industrial seedling process of the Zealand chrysanthemum can be effectively shortened, the propagation is rapid, the transplanting survival rate is high, the pollution rate is low, and the comprehensive cost is low.
In summary, the invention includes at least one of the following beneficial technical effects:
1. the invention provides an antibacterial rooting medium special for Zea mays and a preparation method thereof, the formula is simple, the preparation method is simple, the cost is low, high-temperature disinfection and pollution resistance are avoided, the growth period in the seedling cultivation process is short, only 20-25 days are needed, the propagation is rapid, the rooting can be realized in 5-7 days, the rooting rate is 100%, the seedling rate is 96-100%, and the 3-day transplanting survival rate is 98-100%;
2. the invention provides a rooting culture method, which can directly carry out rooting and seedling exercising in a natural environment of a plastic greenhouse through sealing a preservative film and controlling environmental factors in the tissue culture process, greatly reduce the pollution rate during rooting and seedling exercising and reduce the seedling exercising time.
Detailed Description
The present invention is further explained with reference to the following specific examples, but it should be noted that the following examples are only illustrative of the present invention and should not be construed as limiting the present invention, and all technical solutions similar or equivalent to the present invention are within the scope of the present invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art, and the raw materials used are commercially available products.
In the following examples and comparative examples, ferrous sulfate was sourced from Guangzhou chemical agent works/Guangzhou brand; the disodium salt of ethylene diamine tetraacetic acid is from Guangzhou chemical reagent factory/Guangzhou brand; boric acid was sourced from guangzhou chemical reagent house/guangzhou brand; the alpha-naphthylacetic acid is from Shanghai-sourced Biotechnology Co., Ltd./YUAN JU; the indolebutyric acid is from Shanghai-sourced bioscience Co., Ltd./YUAN JU; the nano silver solution was obtained from materials science and technology ltd of Guangzhou City (silver ion concentration of the solution was 100ppm), and the type of nano silver in the nano silver solution was nano silver B in the following examples and comparative examples; penicillin is derived from Ruiyang pharmaceutical Co., Ltd; streptomycin is derived from regiang pharmaceuticals, inc.
Example 1
The antibacterial rooting culture medium special for the Zealand chrysanthemum comprises the following raw materials in parts by weight:
600mg/L of ammonia nitrate; potassium nitrate is 800 mg/L; 100mg/L potassium dihydrogen phosphate; 250mg/L of calcium chloride; magnesium sulfate is 100 mg/L; 20mg/L of ferrous sulfate; ethylenediaminetetraacetic acid disodium salt 30 mg/L; 0.1mg/L of sodium mesh; 10mg/L of manganese sulfate; 5mg/L of zinc sulfate; copper sulfate 0.01 mg/L; cobalt chloride 0.01 mg/L; boric acid 4 mg/L; a-naphthylacetic acid 0.05 mg/L; 0.1mg/L of indolebutyric acid; 10ml of nano silver solution; 10000 units of penicillin; 8000 units of streptomycin and 2g/L of carrageenan.
A preparation method of an antibacterial rooting culture medium special for Zealand chrysanthemum comprises the following steps:
s1, adding ammonia nitrate, potassium nitrate, monopotassium phosphate, calcium chloride, magnesium sulfate, ferrous sulfate and ethylene diamine tetraacetic acid disodium salt in the weight ratio into a stainless steel pot, and stirring and mixing to form a mixed solution A, wherein the stirring speed is 200rpm/min, and the stirring time is 20 min;
s2, stirring and mixing the nano-silver solution, the penicillin and the streptomycin in the weight ratio to form a mixed solution B;
s3, raising the temperature of the mixed solution A to 80 ℃, adding the sodium mesh, the manganese sulfate, the zinc sulfate, the copper sulfate, the cobalt chloride, the boric acid, the alpha-naphthylacetic acid, the indolebutyric acid and the carrageenan in the weight ratio, stirring and mixing until the carrageenan is completely dissolved to form a mixed solution C;
s4, reducing the temperature of the mixed solution C to 60 ℃, adding the mixed solution B into the mixed solution C, uniformly stirring, and adjusting the pH value to 4.2 to obtain the antibacterial rooting culture medium special for the Zealand chrysanthemum.
In this example, the weight percentage of the nano silver solution is 1 wt%.
Example 2
The antibacterial rooting culture medium special for the Zealand chrysanthemum comprises the following raw materials in parts by weight:
900mg/L of ammonia nitrate; potassium nitrate 1000 mg/L; 150mg/L of monopotassium phosphate; calcium chloride 200 mg/L; magnesium sulfate 150 mg/L; 25mg/L of ferrous sulfate; 35mg/L of ethylene diamine tetraacetic acid disodium salt; 0.2mg/L of sodium mesh; 20mg/L of manganese sulfate; zinc sulfate 8 mg/L; copper sulfate 0.02 mg/L; cobalt chloride 0.02 mg/L; boric acid 5 mg/L; a-naphthylacetic acid 0.1 mg/L; 0.3mg/L of indolebutyric acid; 20ml of nano silver solution; penicillin 15000 units; 10000 units of streptomycin and 4g/L of carrageenan.
A preparation method of an antibacterial rooting culture medium special for Zealand chrysanthemum comprises the following steps:
s1, adding ammonia nitrate, potassium nitrate, monopotassium phosphate, calcium chloride, magnesium sulfate, ferrous sulfate and ethylene diamine tetraacetic acid disodium salt in the weight ratio into a stainless steel pot, and stirring and mixing to form a mixed solution A, wherein the stirring speed is 250rpm/min, and the stirring time is 25 min;
s2, stirring and mixing the nano-silver solution, the penicillin and the streptomycin in the weight ratio to form a mixed solution B;
s3, raising the temperature of the mixed solution A to 80 ℃, adding the sodium mesh, the manganese sulfate, the zinc sulfate, the copper sulfate, the cobalt chloride, the boric acid, the alpha-naphthylacetic acid, the indolebutyric acid and the carrageenan in the weight ratio, stirring and mixing until the carrageenan is completely dissolved to form a mixed solution C;
s4, reducing the temperature of the mixed solution C to 65 ℃, adding the mixed solution B into the mixed solution C, uniformly stirring, and adjusting the pH value to 4.8 to obtain the special antibacterial rooting culture medium for the Zealand chrysanthemum.
In this example, the weight percentage of the nano silver solution is 2 wt%.
Example 3
The antibacterial rooting culture medium special for the Zealand chrysanthemum comprises the following raw materials in parts by weight:
800mg/L of ammonia nitrate; potassium nitrate is 1100 mg/L; potassium dihydrogen phosphate 175 mg/L; 280mg/L of calcium chloride; magnesium sulfate is 200 mg/L; 25mg/L of ferrous sulfate; ethylene diamine tetraacetic acid disodium salt 38 mg/L; 0.4mg/L of sodium mesh; 25mg/L of manganese sulfate; zinc sulfate 12 mg/L; copper sulfate 0.025 mg/L; cobalt chloride 0.025 mg/L; boric acid 6 mg/L; a-naphthylacetic acid 0.15 mg/L; 0.6mg/L of indolebutyric acid; 30ml of nano silver solution; penicillin 18000 units; 12000 units of streptomycin and 6g/L of carrageenan.
A preparation method of an antibacterial rooting culture medium special for Zealand chrysanthemum comprises the following steps:
s1, adding ammonia nitrate, potassium nitrate, monopotassium phosphate, calcium chloride, magnesium sulfate, ferrous sulfate and ethylene diamine tetraacetic acid disodium salt in the weight ratio into a stainless steel pot, and stirring and mixing to form a mixed solution A, wherein the stirring speed is 300rpm/min, and the stirring time is 25 min;
s2, stirring and mixing the nano-silver solution, the penicillin and the streptomycin in the weight ratio to form a mixed solution B;
s3, raising the temperature of the mixed solution A to 85 ℃, adding the sodium mesh, the manganese sulfate, the zinc sulfate, the copper sulfate, the cobalt chloride, the boric acid, the alpha-naphthylacetic acid, the indolebutyric acid and the carrageenan in the weight ratio, stirring and mixing until the carrageenan is completely dissolved to form a mixed solution C;
s4, reducing the temperature of the mixed solution C to 70 ℃, adding the mixed solution B into the mixed solution C, uniformly stirring, and adjusting the pH value to 5.4 to obtain the antibacterial rooting culture medium special for the Zealand chrysanthemum.
In this example, the weight percentage of the nano silver solution is 3 wt%.
Example 4
The antibacterial rooting culture medium special for the Zealand chrysanthemum comprises the following raw materials in parts by weight:
1000mg/L of ammonium nitrate; potassium nitrate 1200 mg/L; potassium dihydrogen phosphate is 200 mg/L; 300mg/L of calcium chloride; magnesium sulfate 250 mg/L; 30mg/L of ferrous sulfate; ethylene diamine tetraacetic acid disodium salt 40 mg/L; 0.5mg/L of sodium mesh; 30mg/L of manganese sulfate; zinc sulfate 15 mg/L; copper sulfate 0.03 mg/L; cobalt chloride 0.03 mg/L; boric acid 8 mg/L; a-naphthylacetic acid 0.2 mg/L; 1mg/L of indolebutyric acid; 40ml of nano silver solution; penicillin 20000 units; 14000 units of streptomycin and 10g/L of carrageenan.
A preparation method of an antibacterial rooting culture medium special for Zealand chrysanthemum comprises the following steps:
s1, adding ammonia nitrate, potassium nitrate, monopotassium phosphate, calcium chloride, magnesium sulfate, ferrous sulfate and ethylene diamine tetraacetic acid disodium salt in the weight ratio into a stainless steel pot, and stirring and mixing to form a mixed solution A, wherein the stirring speed is 350rpm/min, and the stirring time is 30 min;
s2, stirring and mixing the nano-silver solution, the penicillin and the streptomycin in the weight ratio to form a mixed solution B;
s3, raising the temperature of the mixed solution A to 90 ℃, adding the sodium mesh, the manganese sulfate, the zinc sulfate, the copper sulfate, the cobalt chloride, the boric acid, the alpha-naphthylacetic acid, the indolebutyric acid and the carrageenan in the weight ratio, stirring and mixing until the carrageenan is completely dissolved to form a mixed solution C;
and S4, reducing the temperature of the mixed solution C to 70 ℃, adding the mixed solution B into the mixed solution C, uniformly stirring, and adjusting the pH value to 6.3 to obtain the antibacterial rooting culture medium special for the Zealand chrysanthemum.
In this example, the weight percentage of the nano silver solution is 4 wt%.
Example 5
The antibacterial rooting culture medium special for the Zealand chrysanthemum comprises the following raw materials in parts by weight:
825mg/L of ammonia nitrate; potassium nitrate 950 mg/L; 185mg/L of monopotassium phosphate; 220mg/L of calcium chloride; 185mg/L magnesium sulfate; 27.8mg/L ferrous sulfate; ethylenediaminetetraacetic acid disodium salt 37.3 mg/L; 0.25mg/L of sodium mesh; manganese sulfate 22.3 mg/L; zinc sulfate 8.6 mg/L; copper sulfate 0.025 mg/L; cobalt chloride 0.025 mg/L; boric acid 6.2 mg/L; a-naphthylacetic acid 0.1 mg/L; 0.4mg/L of indolebutyric acid; 20ml of nano silver solution; penicillin 16000 units; 10000 units of streptomycin and 6g/L of carrageenan.
A preparation method of an antibacterial rooting culture medium special for Zealand chrysanthemum comprises the following steps:
s1, adding ammonia nitrate, potassium nitrate, monopotassium phosphate, calcium chloride, magnesium sulfate, ferrous sulfate and ethylene diamine tetraacetic acid disodium salt in the weight ratio into a stainless steel pot, and stirring and mixing to form a mixed solution A, wherein the stirring speed is 300rpm/min, and the stirring time is 25 min;
s2, stirring and mixing the nano-silver solution, the penicillin and the streptomycin in the weight ratio to form a mixed solution B;
s3, raising the temperature of the mixed solution A to 90 ℃, adding the sodium mesh, the manganese sulfate, the zinc sulfate, the copper sulfate, the cobalt chloride, the boric acid, the alpha-naphthylacetic acid, the indolebutyric acid and the carrageenan in the weight ratio, stirring and mixing until the carrageenan is completely dissolved to form a mixed solution C;
s4, reducing the temperature of the mixed solution C to 65 ℃, adding the mixed solution B into the mixed solution C, uniformly stirring, and adjusting the pH value to 5.8 to obtain the special antibacterial rooting culture medium for the Zealand chrysanthemum.
In this example, the weight percentage of the nano silver solution is 2 wt%.
Example 6
A rooting culture method of Zealand chrysanthemum comprises the following steps:
B. taking tender shoots in the excellent-shape subculture propagation material of the Zealand chrysanthemum as an inoculation material, inoculating the tender shoots into the special antibacterial rooting culture medium for the Zealand chrysanthemum prepared in the embodiment 1-5, and culturing to obtain chrysanthemum tissue culture seedlings, wrapping the chrysanthemum tissue culture seedlings for 2-3 circles along the joint of the bottle cap and the bottle by using a food-grade preservative film, and keeping the bottle cap and the bottle to be wrapped by 1-1.5 cm at two sides respectively;
B. after the tissue culture seedlings of the chrysanthemum are sealed by a preservative film and at the joint of a bottle and a bottle, the tissue culture seedlings are placed on a seedling hardening plastic greenhouse internal frame for rooting and seedling hardening culture at the temperature of 15-35 ℃, the illumination intensity of 8000-15000 Lux, the illumination time of 7-11 h and the humidity of 35-85% every day, the rooting and seedling hardening culture is carried out for 20-25 d, and the seedlings can be transplanted when the roots are tidy and the roots are 3-7 cm high.
Comparative example 1
Compared with example 5, the difference is only that the nano silver solution and penicillin are absent, and the rest parameters and the preparation method are the same as example 5.
Comparative example 2
Compared with example 5, the difference is only that the nano silver solution is absent, and the rest parameters and the preparation method are the same as example 5.
Comparative example 3
Compared with example 5, the difference is only that the penicillin is replaced by the same unit of streptomycin, namely the unit of streptomycin is 26000, and the rest parameters are the same as the preparation method of example 5.
Comparative example 4
The difference compared to example 5 is only the absence of penicillin and streptomycin, and the remaining parameters are the same as for example 5.
Test examples, comparative tests
Raw materials: the antibacterial rooting culture medium special for the Zealand chrysanthemum is prepared in the examples 1-5 and the comparative examples 1-3; a control group prepared by MS + NAA + sucrose and sterilized by high temperature and moist heat (121 ℃, 20min for sterilization) and a control group 1 and a control group 2 which is not sterilized; (ii) a
The test method comprises the following steps: adopting the inoculation method of the embodiment 6 to respectively inoculate the tender shoots in the same subculture proliferation material of the Zephyranthes roseus as the inoculation material into the special antibacterial rooting culture medium for the Zephyranthes roseus prepared in the embodiments 1-5 and the comparative examples 1-3 for culture, sealing the bottle cap and the bottle interface by a preservative film, then placing the bottle cap and the bottle interface on a seedling hardening plastic greenhouse inner frame for rooting and seedling hardening culture at the temperature of 15-35 ℃, the illumination intensity of 8000-15000 Lux, the illumination time of 7-11 h and the humidity of 35-85% every day, and transplanting the seedling after rooting and seedling hardening culture for 20 days, wherein the seedling is in order in rooting and the root is 3-7 cm high;
adopting the tender shoots in the same subculture multiplication material of the Zealand chrysanthemum as an inoculation material to inoculate in the culture media of the control group 1 and the control group 2, culturing the inoculated tender shoots in a culture room for 25 days, and then hardening seedlings in a hardening plastic greenhouse for 20 days; after culturing for 1 month, recording the growth cycle, rooting speed, rooting rate, bud seedling growth condition, pollution rate, seedling rate and 30-day transplanting survival rate of the Zealand chrysanthemum; the results are shown in Table 1.
TABLE 1
It is worth mentioning that: the sterilization and disinfection of the culture medium mainly comprises a nano-silver solution, penicillin and streptavidin are added to achieve the effect-enhancing synergistic effect, the nano-silver solution is absent, the sterilization and disinfection effects of penicillin and streptomycin are only small, the common culture medium is subjected to high-temperature damp-heat disinfection (121 ℃, and sterilization is carried out for 20 min), otherwise, the common culture medium is completely polluted in 3-7 days, and the material is completely polluted and dies slowly.
According to the data in the table 1, the seedlings cultured by the antibacterial rooting culture medium special for the Zealand chrysanthemum are strong, emerald green in color and regular in rooting, the roots are 4-5 cm high, the seedlings grown by the culture medium of the control group are light green in color, and the roots of the seedlings are uneven and 5-7 cm high; the growth cycle of the seedling cultured by the antibacterial rooting medium special for the Zephyranthes candida prepared in the embodiment 1-5 of the invention is 20-25 days, the seedling can root in 5-7 days, the rooting rate can be improved to 100%, the seedling rate is 96-100%, and the 3-day transplanting survival rate is 98-100%, wherein the embodiment 5 is the best embodiment, the growth cycle of the seedling cultured by the medium of the control group is longer, 35-40 days, the time cost is higher, and the effects of the rooting speed, the rooting rate, the seedling rate and the 30-day transplanting survival rate are inferior to those of the embodiment 1-5.
Compared with the example 5, the comparative example 1 lacks the nano silver solution and the penicillin, the growth cycle of the nano silver solution and the growth cycle of the penicillin are the same as the growth cycle of a control group, the pollution rate is high, indexes such as the rooting speed, the rooting rate, the seedling rate and the 30-day transplanting survival rate are all reduced, the influence of the addition of the nano silver solution and the penicillin on the indexes is proved, the pollution rate is particularly greatly influenced, the pollution rate reaches 95%, and the nano silver solution and the penicillin are added to act together with other components in a culture medium, so that the indexes such as the rooting speed, the rooting rate, the pollution rate, the seedling rate and the 30-day transplanting survival rate can be better;
the culture medium prepared in the comparative example 2 lacks the nano-silver solution, the effect of indexes such as rooting rate, seedling rate and transplanting survival rate is better than that of the culture medium prepared in the comparative example 1, but the pollution rate is greatly influenced, the pollution rate reaches 96 percent, the effect is worse than that of the culture medium prepared in the examples 1 to 5, particularly, the compounding of the nano-silver solution and penicillin can effectively reduce the growth cycle and the pollution rate of seedlings and can effectively improve the rooting rate, the seedling rate and the 30-day transplanting survival rate;
the effect of the indexes of the nano-silver solution is better than that of the index of the nano-silver solution in comparative example 3 or comparative example 4, but the nano-silver solution is poorer than that of the nano-silver solution in examples 1-5, so that the compounding of the nano-silver solution and the penicillin can effectively reduce the growth cycle and the pollution rate of the seedling and can effectively improve the rooting rate, the seedling rate and the 30-day transplanting survival rate; comparative example 3 lacks penicillin, but in the presence of streptomycin, the nano-silver solution and streptomycin are compounded, so that a good effect cannot be obtained, and the rooting rate, the seedling rate and the 30-day transplanting survival rate of the nano-silver solution are not good as the index effects of examples 1-5. Comparative example 3 or comparative example 4 shows that the sterilization and disinfection of the culture medium are mainly realized by the nano-silver solution, the addition of the penicillin and the streptomycin has synergistic effect, the nano-silver solution is lacked, and the sterilization and disinfection effects of only the penicillin and the streptomycin are not great.
According to the invention, organic matters and sucrose in the MS culture medium are removed innovatively, so that the culture medium is simplified, the cost of the culture medium is greatly reduced, the preparation process of the culture medium is simplified, and the risk of pollution caused by the organic matters and the sucrose is reduced; and then adding bactericides such as nano-silver solution, penicillin, streptomycin and the like and antibiotics, so that the pollution rate of the culture medium can be effectively controlled without high-temperature sterilization, and the pollution rate is reduced to be below 3%.
The foregoing embodiments are merely illustrative of the principles and utilities of the present invention and are not intended to limit the invention. Any person skilled in the art can modify or change the above-mentioned embodiments without departing from the spirit and scope of the present invention. Accordingly, it is intended that all equivalent modifications or changes which can be made by those skilled in the art without departing from the spirit and technical spirit of the present invention be covered by the claims of the present invention.
Claims (9)
1. The antibacterial rooting culture medium special for Zealand chrysanthemum is characterized by comprising the following raw materials in parts by weight:
1800-2950 mg/L macroelements; 20-30 mg/L of ferrous sulfate; 30-40 mg/L of ethylene diamine tetraacetic acid disodium salt; 0.1-0.5 mg/L sodium mesh; 10-30 mg/L of manganese sulfate; 5-15 mg/L of zinc sulfate; 0.01-0.03 mg/L of copper sulfate; 0.01-0.03 mg/L of cobalt chloride; boric acid 4-8 mg/L; 0.05-0.2 mg/L of alpha-naphthylacetic acid; 0.1-1 mg/L of indolebutyric acid; 10-40 ml of nano silver solution; 10000-20000 units of penicillin; 8000-14000 units of streptomycin and 2-10 g/L of coagulant.
2. The special antibacterial rooting culture medium for enriched chrysanthemum according to claim 1, which is characterized by comprising the following raw materials in parts by weight:
2400-2950 mg/L macroelements; 25-30 mg/L of ferrous sulfate; 35-40 mg/L of ethylene diamine tetraacetic acid disodium salt; 0.2-0.5 mg/L of sodium mesh; 20-30 mg/L of manganese sulfate; 8-15 mg/L of zinc sulfate; 0.02-0.03 mg/L of copper sulfate; 0.02-0.03 mg/L of cobalt chloride; 5-8 mg/L of boric acid; 0.05-0.15 mg/L of alpha-naphthylacetic acid; 0.3-1 mg/L of indolebutyric acid; 10-30 ml of nano silver solution; 15000-20000 units of penicillin; 8000-12000 units of streptomycin and 4-10 g/L of coagulant.
3. The antibacterial rooting culture medium special for enriched chrysanthemum according to any one of claims 1-2, wherein the weight percentage of the nano silver solution is 1-4 wt%.
4. The antibacterial rooting medium special for enriched chrysanthemum according to claim 3, wherein the weight percentage of the nano silver solution is 2 wt%.
5. The antibacterial rooting medium special for coreopsis tinctoria according to any one of claims 1-2, wherein the macroelements comprise ammonium nitrate, potassium dihydrogen phosphate, calcium chloride and magnesium sulfate.
6. The antibacterial rooting medium special for coreopsis tinctoria according to any one of claims 1-2, wherein the coagulant comprises carrageenan.
7. A preparation method of an antibacterial rooting culture medium special for Zealand chrysanthemum is characterized by comprising the following steps:
s1, stirring and mixing the major elements, ferrous sulfate and ethylene diamine tetraacetic acid disodium salt according to the weight ratio to form a mixed solution A;
s2, stirring and mixing the nano-silver solution, the penicillin and the streptomycin in the weight ratio to form a mixed solution B;
s3, raising the temperature of the mixed solution A to 80-90 ℃, adding sodium mesh, manganese sulfate, zinc sulfate, copper sulfate, cobalt chloride, boric acid, alpha-naphthylacetic acid, indolebutyric acid and a coagulant in the weight ratio, stirring and mixing until carrageenan is completely dissolved to form a mixed solution C;
and S4, reducing the temperature of the mixed solution C to 60-70 ℃, adding the mixed solution B into the mixed solution C, uniformly stirring, and adjusting the pH value to 4.2-6.3 to obtain the antibacterial rooting culture medium special for the Zea mays.
8. The preparation method of the antibacterial rooting medium special for Dendranthema giganteum according to claim 7, wherein the pH value of the antibacterial rooting medium special for Dendranthema giganteum is 5.8.
9. A rooting culture method of Zealand chrysanthemum is characterized by comprising the following steps:
A. taking tender shoots in the excellent-shape subculture proliferation material of the Zealand chrysanthemum as an inoculation material to inoculate in the special antibacterial rooting culture medium for the Zealand chrysanthemum according to any one of claims 1 to 6, culturing to obtain chrysanthemum tissue culture seedlings, binding the chrysanthemum tissue culture seedlings for 2 to 3 circles along the joint of the bottle cap and the bottle by using a food-grade preservative film, and keeping the bottle cap and the bottle to be wrapped by 1 to 1.5cm at two sides respectively;
B. after the tissue culture seedlings of the chrysanthemum are sealed by a preservative film and at the joint of a bottle and a bottle, the tissue culture seedlings are placed on a seedling hardening plastic greenhouse internal frame for rooting and seedling hardening culture at the temperature of 15-35 ℃, the illumination intensity of 8000-15000 Lux, the illumination time of 7-11 h and the humidity of 35-85% every day, the rooting and seedling hardening culture is carried out for 20-25 d, and the seedlings can be transplanted when the roots are tidy and the roots are 3-7 cm high.
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