CN114431143A - Improved MS culture medium and preparation method and application thereof - Google Patents
Improved MS culture medium and preparation method and application thereof Download PDFInfo
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- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
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Abstract
The invention relates to an improved MS culture medium and a preparation method and application thereof, belonging to the technical field of tissue culture. The improved MS culture medium comprises the following components: ammonium sulfate, potassium nitrate, 7-water magnesium sulfate, monopotassium phosphate, calcium chloride, manganese sulfate, 7-water zinc sulfate, boric acid, potassium iodide, sodium molybdate, copper sulfate, cobalt chloride, 7-water ferric sulfate, disodium ethylene diamine tetraacetate, glycine, thiamine hydrochloride, pyridoxine hydrochloride, inositol, indoleacetic acid, 6-benzylaminopurine, carrageenan and xanthan gum. Compared with the traditional induced differentiation medium, the induced differentiation medium can obviously improve the seedling rate and the differentiation rate of the stem tip meristem of the sweet potato, and provides a basis for the rapid propagation and the large-scale production of the sweet potato.
Description
Technical Field
The invention relates to the technical field of tissue culture, in particular to an improved MS culture medium and a preparation method and application thereof.
Background
Sweet potatoes are receiving more and more attention from people as cash crops integrating the functions of energy, food processing, health care, medicine, export foreign exchange and the like. Sweet potato is a kind of heterosis crop, and the propagation of sweet potato virus disease is always caused by adopting vegetative propagation, so that the yield and quality are reduced. The MS culture medium is designed for tobacco cell culture in 1962 by Murashige and Skoog, and is characterized by higher inorganic salt and ion concentration, more stable ion balance solution, high nitrate content, proper nutrient quantity and proportion, and capability of meeting the nutritional and physiological requirements of plant cells, so the MS culture medium is often used as a culture medium for sweet potato tissue culture. However, when the stem tip meristem is cultured by the traditional MS culture medium in the sweet potato tissue, the differentiation efficiency is low, the growth period is long, and the traditional MS culture medium is not suitable for the rapid propagation and large-scale production of the sweet potato. Therefore, it is an urgent need to solve the problem of obtaining an improved MS culture medium with high differentiation efficiency and short growth cycle.
Disclosure of Invention
The invention aims to provide an improved MS culture medium and a preparation method and application thereof.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides an improved MS culture medium, which takes water as a solvent, and each liter of the improved MS culture medium comprises the following components by mass:
1.3 to 1.34g of ammonium sulfate, 1.8 to 2.2g of potassium nitrate, 0.35 to 0.39g of 7-water magnesium sulfate, 0.16 to 0.18g of monopotassium phosphate, 0.42 to 0.46g of calcium chloride, 0.0167 to 0.0171g of manganese sulfate, 0.0082 to 0.0090g of 7-water zinc sulfate, 0.0062 to 0.0066g of boric acid, 0.00080 to 0.00086g of potassium iodide, 0.00023 to 0.00027g of sodium molybdate, 0.000023 to 0.000027g of copper sulfate, 0.000023 to 0.000027g of cobalt chloride, 0.0276 to 0.0280g of 7-water ferric sulfate, 0.0371 to 0.0375g of disodium ethylenediaminetetraacetate, 0.0018 to 0.0022g of glycine, 0.0003 to 0.0005g of thiamine hydrochloride, 0.0004 to 0.0006g of pyridoxine hydrochloride, 0.08 to 0.08g of carrageenan, 0.0004 to 0.0004g of indole acetate, 0.0005 to 0.5 g of xanthan gum, and 0.0005 to 5g of amino acid.
Preferably, water is used as a solvent, and each liter of the improved MS culture medium comprises the following components in mass:
1.31-1.33 g of ammonium sulfate, 1.9-2.1 g of potassium nitrate, 0.36-0.38 g of 7-water magnesium sulfate, 0.165-0.175 g of monopotassium phosphate, 0.43-0.45 g of calcium chloride, 0.0168-0.0170 g of manganese sulfate, 0.0084-0.0088 g of 7-water zinc sulfate, 0.0063-0.0065 g of boric acid, 0.00082-0.00084 g of potassium iodide, 0.00024-0.00026 g of sodium molybdate, 0.000024-0.000026 g of copper sulfate, 0.000024-0.000026 g of cobalt chloride, 0.0277-0.0279 g of 7-water ferric sulfate, 0.0372-0.0374 g of disodium ethylenediaminetetraacetate, 0.0019-0.0021 g of glycine, 0.00035-0.00045 g of thiamine hydrochloride, 0.00045-0.00055 g of pyridoxine hydrochloride, 0.09-0.11 g of inositol, 0.45-0.0006.0006 g of indoleacetic acid, 0.0006-0.00035 g of carrageenan, and 2.0008-2.8 g of benzyl amino acid.
Preferably, water is used as a solvent, and each liter of the improved MS culture medium comprises the following components in mass:
1.32g of ammonium sulfate, 2g of potassium nitrate, 0.37g of 7-hydrate magnesium sulfate, 0.17g of monopotassium phosphate, 0.44g of calcium chloride, 0.0169g of manganese sulfate, 0.0086g of 7-hydrate zinc sulfate, 0.0064g of boric acid, 0.00083g of potassium iodide, 0.00025g of sodium molybdate, 0.000025g of copper sulfate, 0.000025g of cobalt chloride, 0.0278g of 7-hydrate ferric sulfate, 0.0373g of disodium ethylenediamine tetraacetate, 0.0020g of glycine, 0.0004g of thiamine hydrochloride, 0.0005g of pyridoxine hydrochloride, 0.1g of inositol, 0.0005g of indoleacetic acid, 0.001g of 6-benzylaminopurine, 4g of carrageenan and 2g of xanthan gum.
Preferably, the pH of the modified MS culture medium is 5.8-6.0.
The invention also provides a preparation method of the improved MS culture medium, which comprises the following steps:
(1) dissolving components except carrageenan and xanthan gum in water to obtain a liquid MS culture medium;
(2) adding carrageenan and xanthan gum into the liquid MS culture medium, and sterilizing for 15-20 min under the conditions of 0.1-0.15 MPa and 118-121 ℃ to obtain the improved MS culture medium.
The invention also provides application of the improved MS culture medium in sweet potato tissue culture.
Preferably, the sweet potato tissue is a sweet potato shoot apical meristem.
Preferably, the tissue culture method comprises the following steps: inoculating the shoot apical meristem of the sweet potato into the modified MS culture medium for culture.
Preferably, the size of the shoot apical meristem of the sweet potato is 0.1 to 0.3 mm.
Preferably, the temperature of the culture is 26-28 ℃, the illumination intensity of the culture is 2800-3200 Lux, the illumination time per day is 12-14 h, and the culture time is 18-22 d.
The invention provides an improved MS culture medium and a preparation method and application thereof. The improved MS culture medium has the following advantages:
(1) the improved MS culture medium is safe, and the ammonium sulfate is used for replacing the ammonium nitrate which is a dangerous chemical easy to explode in the improved MS culture medium, so that the culture medium is safer;
(2) the auxin combination indoleacetic acid and 6-benzylaminopurine which are suitable for the differentiation of the meristematic tissue of the sweet potato stem apex are added into the improved MS culture medium, so that the differentiation of the meristematic tissue of the sweet potato stem apex is promoted, and the differentiation efficiency and the differentiation speed are improved;
(3) the culture medium optimizes the curing agent components, and combines carrageenan suitable for the meristem differentiation of sweet potatoes and xanthan gum to be more suitable for the growth of sweet potatoes than agar.
Compared with the traditional MS culture medium, the improved MS culture medium is more suitable for culturing the stem tip tissue of the sweet potato, and can improve the differentiation efficiency by at least 1.9 times.
Detailed Description
The invention provides an improved MS culture medium, which takes water as a solvent, and each liter of the improved MS culture medium comprises the following components by mass:
1.3 to 1.34g of ammonium sulfate, preferably 1.31 to 1.33g, and more preferably 1.32 g;
1.8-2.2 g of potassium nitrate, preferably 1.9-2.1 g, and more preferably 2 g;
7, 0.35 to 0.39g of magnesium sulfate hydrate, preferably 0.36 to 0.38g, and more preferably 0.37 g;
0.16-0.18 g of monopotassium phosphate, preferably 0.165-0.175 g, and more preferably 0.17 g;
0.42-0.46 g of calcium chloride, preferably 0.43-0.45 g, and more preferably 0.44 g;
0.0167-0.0171 g of manganese sulfate, preferably 0.0168-0.0170 g, and more preferably 0.0169 g;
0.0082-0.0090 g of zinc sulfate hydrate, preferably 0.0084-0.0088 g, and further preferably 0.0086 g;
0.0062-0.0066 g of boric acid, preferably 0.0063-0.0065 g, and more preferably 0.0064 g;
0.00080 to 0.00086g of potassium iodide, preferably 0.00082 to 0.00084g, and more preferably 0.00083 g;
0.00023-0.00027 g of sodium molybdate, preferably 0.00024-0.00026 g of sodium molybdate, and more preferably 0.00025g of sodium molybdate;
0.000023-0.000027 g of copper sulfate, preferably 0.000024-0.000026 g of copper sulfate, and further preferably 0.000025g of copper sulfate;
0.000023-0.000027 g of cobalt chloride, preferably 0.000024-0.000026 g, and more preferably 0.000025 g;
0.0276-0.0280 g of ferric sulfate hydrate, preferably 0.0277-0.0279 g, and further preferably 0.0278 g;
0.0371-0.0375 g of disodium ethylene diamine tetraacetate, preferably 0.0372-0.0374 g, and more preferably 0.0373 g;
0.0018-0.0022 g of glycine, preferably 0.0019-0.0021 g, and further preferably 0.0020 g;
thiamine hydrochloride in an amount of 0.0003 to 0.0005g, preferably 0.00035 to 0.00045g, and more preferably 0.0004 g;
pyridoxine hydrochloride in an amount of 0.0004 to 0.0006g, preferably 0.00045 to 0.00055g, and more preferably 0.0005 g;
0.08-0.12 g of inositol, preferably 0.09-0.11 g, and more preferably 0.1 g;
indoleacetic acid 0.0004-0.0006 g, preferably 0.00045-0.00055 g, and more preferably 0.0005 g;
6-benzylaminopurine 0.0005-0.0015 g, preferably 0.00075-0.00125 g, and more preferably 0.001 g;
3.5-4.5 g of carrageenan, preferably 3.8-4.2 g, and more preferably 4 g;
1.5 to 2.5g of xanthan gum, preferably 1.8 to 2.2g, and more preferably 2 g.
In the invention, the pH of the modified MS culture medium is 5.8-6.0, and is preferably 5.9.
In the present invention, the agent for adjusting the pH of the modified MS medium is hydrochloric acid or sodium hydroxide.
The invention also provides a preparation method of the improved MS culture medium, which comprises the following steps:
(1) dissolving components except carrageenan and xanthan gum in water to obtain a liquid MS culture medium;
(2) adding carrageenan and xanthan gum into the liquid MS culture medium, and sterilizing for 15-20 min under the conditions of 0.1-0.15 MPa and 118-121 ℃ to obtain the improved MS culture medium.
In the present invention, the pressure of the sterilization is preferably 0.125 MPa; the sterilization temperature is preferably 119-120 ℃, and is further preferably 119.5 ℃; the time for sterilization is preferably 17-18 min, and more preferably 17.5 min.
The invention also provides application of the improved MS culture medium in sweet potato tissue culture.
In the present invention, the sweetpotato tissue is a sweetpotato shoot apical meristem.
In the invention, the tissue culture method comprises the following steps: inoculating the shoot apical meristem of the sweet potato into the modified MS culture medium for culture.
In the present invention, the shoot apical meristem is obtained by a method comprising:
selecting a sweet potato plant which grows well and has plump stem tip, taking 2cm of apical bud, washing 2-3 times with water for later use, soaking the stem tip in 75% alcohol for 30s in an aseptic environment, then disinfecting with 2% hypochlorous acid solution for 5min, and soaking in aseptic water for later use. And (3) in an aseptic environment, placing the stem tip under a dissecting mirror, peeling off the apical bud or the bigger young leaf on the axillary bud, and cutting a stem tip tissue with the diameter of 0.1-0.3 mm to obtain a stem tip meristem.
In the present invention, each of the shoot apical meristems carries 1 leaf primordia.
In the invention, the size of the shoot apical meristem of the sweet potato is 0.1-0.3 mm, preferably 0.2 mm.
In the invention, the culture temperature is 26-28 ℃, and preferably 27 ℃;
the illumination intensity of the culture is 2800-3200 Lux, and 3000Lux is preferred;
the illumination time per day is 12-14 h, preferably 13 h;
the culture time is 18-22 days, and preferably 20 days.
The embodiments of the present invention will be described in detail with reference to the following examples, but they should not be construed as limiting the scope of the present invention.
The seedling formation rate in the examples of the present invention and the comparative examples is (number of seedlings formed/number of meristems of shoot tips inoculated) × 100%.
In the examples of the present invention and comparative examples, the differentiation rate (the number of total buds differentiated/the number of meristems of shoot tips inoculated) x 100% was described.
The sweet potato variety used in the examples of the present invention and the comparative examples was commercial potato 19.
Example 1
1.3g of ammonium sulfate, 2.0g of potassium nitrate, 0.39g of 7-water magnesium sulfate, 0.17g of monopotassium phosphate, 0.42g of calcium chloride, 0.0171g of manganese sulfate, 0.0082g of 7-water zinc sulfate, 0.0065g of boric acid, 0.00086g of potassium iodide, 0.00025g of sodium molybdate, 0.000023g of copper sulfate, 0.000027g of cobalt chloride, 0.0278g of 7-water ferric sulfate, 0.0375g of disodium ethylenediamine tetraacetate, 0.0018g of glycine, 0.0003g of thiamine hydrochloride, 0.0005g of pyridoxine hydrochloride, 0.12g of inositol, 0.0005g of indoleacetic acid and 0.0005g of 6-benzylaminopurine are dissolved in 800mL of water, the dissolved solution is made up to 1000mL of water, and the pH is adjusted to 6.0. Then 4.5g of carrageenan and 1.5g of xanthan gum are added, and after the preparation is finished, the mixture is placed in an autoclave at the temperature of 121 ℃ and the pressure of 0.01MPa for sterilization for 15min, and the improved MS culture medium is obtained.
Selecting a sweet potato plant which grows well and has plump stem tip, taking 2cm of apical bud, washing 2 times with water for later use, wiping and sterilizing with 75% alcohol, then soaking in mercuric chloride solution for 10min, peeling off the apical bud or the bigger young leaf on the axillary bud by placing the stem tip under a dissecting mirror in an aseptic environment, and cutting off 0.2mm of stem tip tissue to obtain the stem tip meristem.
The shoot tip tissue to be inoculated is inoculated in the improved MS culture medium, the seedling formation number is observed after the shoot tip tissue is cultured for 20 days under the conditions that the temperature is 26 ℃, the illumination intensity is 2800Lux and the illumination is 14h every day, the seedling formation rate is calculated, and the result is shown in Table 1. The number of total buds differentiated was observed, and the differentiation rate was calculated, and the results are shown in Table 2.
Example 2
1.34g of ammonium sulfate, 1.8g of potassium nitrate, 0.36g of 7-water magnesium sulfate, 0.18g of monopotassium phosphate, 0.46g of calcium chloride, 0.0167g of manganese sulfate, 0.0090g of 7-water zinc sulfate, 0.0062g of boric acid, 0.0008g of potassium iodide, 0.00027g of sodium molybdate, 0.000025g of copper sulfate, 0.000025g of cobalt chloride, 0.0276g of 7-water ferric sulfate, 0.0371g of disodium ethylenediaminetetraacetate, 0.002g of glycine, 0.0005g of thiamine hydrochloride, 0.0006g of pyridoxine hydrochloride, 0.08g of inositol, 0.0004g of indoleacetic acid and 0.0015g of 6-benzylaminopurine are dissolved in 800mL of water, the dissolved solution is supplemented to 1000mL of water, and the pH is adjusted to 6.0. Then 3.5g of carrageenan and 2.0g of xanthan gum are added, and after the preparation is finished, the mixture is placed in an autoclave at 118 ℃ and 0.015MPa for sterilization for 20min, and the improved MS culture medium is obtained.
Selecting a sweet potato plant which grows well and has plump stem tip, taking 2cm of terminal bud, washing with water for 3 times for later use, wiping and sterilizing with 75% alcohol, then soaking in mercuric chloride solution for 10min, peeling off the larger young leaves on the terminal bud or axillary bud by placing the stem tip under a dissecting mirror in an aseptic environment, and cutting off 0.3mm of stem tip tissue to obtain a stem tip meristem. The shoot apical meristem to be inoculated was obtained.
Inoculating the stem tip tissue to be inoculated into the improved MS culture medium, culturing for 22d under the conditions that the temperature is 28 ℃, the illumination intensity is 3200Lux and the illumination is 12h every day, observing the number of seedlings, and calculating the seedling rate, wherein the result is shown in table 1. The number of total buds differentiated was observed, and the differentiation rate was calculated, and the results are shown in Table 2.
Example 3
1.32g of ammonium sulfate, 2.2g of potassium nitrate, 0.35g of 7-water magnesium sulfate, 0.16g of monopotassium phosphate, 0.45g of calcium chloride, 0.017g of manganese sulfate, 0.0088g of 7-water zinc sulfate, 0.0066g of boric acid, 0.00084g of potassium iodide, 0.00023g of sodium molybdate, 0.000027g of copper sulfate, 0.000023g of cobalt chloride, 0.0280g of 7-water ferric sulfate, 0.0373g of disodium ethylenediamine tetraacetic acid, 0.0022g of glycine, 0.0004g of thiamine hydrochloride, 0.0004g of pyridoxine hydrochloride, 0.1g of inositol, 0.0006g of indoleacetic acid and 0.001g of 6-benzylaminopurine are dissolved in 800mL of water, the solution is made up to 1000mL of water, and the pH is adjusted to 5.9. Then 4g of carrageenan and 2.5g of xanthan gum are added, and after the preparation is finished, the mixture is placed in an autoclave with the temperature of 120 ℃ and the pressure of 0.01MPa for sterilization for 20min, and the improved MS culture medium is obtained.
Selecting a sweet potato plant which grows well and has plump stem tip, taking 2cm of apical bud, washing 2 times with water for later use, wiping and sterilizing with 75% alcohol, then soaking in mercuric chloride solution for 10min, peeling off the apical bud or the bigger young leaf on the axillary bud by placing the stem tip under a dissecting mirror in an aseptic environment, and cutting off 0.2mm of stem tip tissue to obtain the stem tip meristem. Obtaining the stem tip tissue to be inoculated.
The stem tip tissue to be inoculated is inoculated in the improved MS culture medium, the seedling number is observed after the stem tip tissue is cultured for 18 days under the conditions that the temperature is 27 ℃, the illumination intensity is 3000Lux and the illumination is 13 hours per day, the seedling rate is calculated, and the result is shown in Table 1. The number of total buds differentiated was observed, and the differentiation rate was calculated, and the results are shown in Table 2.
Comparative example 1
The scheme of this comparative example 1 was set up according to the method of example 1, and unlike example 1, no indoleacetic acid, 6-benzylaminopurine, and ammonium sulfate were replaced with ammonium nitrate of the same mass, and carrageenan and xanthan gum were replaced with agar of the same mass in the medium of this comparative example 1. After 20 days of culture, the number of seedlings formed was observed, and the rate of formation was calculated, and the results are shown in Table 1. The number of total buds differentiated was observed, and the differentiation rate was calculated, and the results are shown in Table 2.
Comparative example 2
The protocol of this comparative example 2 was set up in the same manner as in example 2, except that in the medium of this comparative example 2, indolacetic acid and 6-benzylaminopurine were replaced with the same mass of sodium a-naphthaleneacetate, and after 22d cultivation, the number of seedlings was observed and the rate of seedlings was calculated, and the results are shown in Table 1. The number of total buds differentiated was observed, and the differentiation rate was calculated, and the results are shown in Table 2.
Comparative example 3
The protocol of comparative example 3 was set up according to the procedure of example 3, and unlike example 3, carrageenan and xanthan gum were replaced with the same mass of agar in the medium of comparative example 3, and after 18d of cultivation, the number of seedlings was observed and the rate of seedlings was calculated, and the results are shown in table 1. The number of total buds differentiated was observed, and the differentiation rate was calculated, and the results are shown in Table 2.
Comparative example 4
The scheme of this comparative example 4 was set up in the same manner as in example 1, except that no indoleacetic acid was added to the medium of this comparative example 4, unlike example 1. After 20 days of culture, the number of seedlings formed was observed, and the rate of formation was calculated, and the results are shown in Table 1. The number of total buds differentiated was observed, and the differentiation rate was calculated, and the results are shown in Table 2.
Comparative example 5
The scheme of this comparative example 5 was set up according to the method of example 1, and unlike example 1, 6-benzylaminopurine was not added to the medium of this comparative example 5. After 20 days of culture, the number of seedlings formed was observed, and the rate of formation was calculated, and the results are shown in Table 1. The number of total buds differentiated was observed, and the differentiation rate was calculated, and the results are shown in Table 2.
TABLE 1 influence of different media on the seedling rate of sweet potatoes
| Group of | Inoculating shoot apical meristem number (number) | Number of adult seedlings | The seedling rate% |
| Example 1 | 40 | 26 | 65 |
| Example 2 | 40 | 30 | 75 |
| Example 3 | 40 | 32 | 80 |
| Comparative example 1 | 40 | 10 | 25 |
| Comparative example 2 | 40 | 16 | 40 |
| Comparative example 3 | 40 | 18 | 45 |
| Comparative example 4 | 40 | 19 | 47.5 |
| Comparative example 5 | 40 | 20 | 50 |
Table 1 shows that the seedling rate of the stem tip meristem of the sweet potato cultured by the culture medium is more than 65 percent, higher than that of the MS culture medium of the traditional comparative example 1, higher than that of the alpha-sodium naphthalene acetate after stimulation and higher than that of the agar after being used as a part of the MS culture medium. The improved MS culture medium can obviously improve the seedling rate of the sweet potatoes.
TABLE 2 Effect of different media on the differentiation Rate of sweetpotato
Table 2 shows that after the culture medium of the invention induces the meristem of the stem tip of the sweet potato, the differentiation rate is obviously higher than that of the culture mediums of comparative examples 1-3 and is more than 1.9 times of that of the culture mediums of comparative examples 1-3.
From the above embodiments, the present invention provides an improved MS culture medium, and a preparation method and application thereof. The improved MS culture medium can obviously improve the seedling rate and the differentiation rate of the meristem of the sweet potato, and provides a basis for batch and large-scale culture of the sweet potato tissues.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (10)
1. An improved MS culture medium is characterized in that water is used as a solvent, and each liter of the improved MS culture medium comprises the following components in mass:
1.3 to 1.34g of ammonium sulfate, 1.8 to 2.2g of potassium nitrate, 0.35 to 0.39g of 7-water magnesium sulfate, 0.16 to 0.18g of monopotassium phosphate, 0.42 to 0.46g of calcium chloride, 0.0167 to 0.0171g of manganese sulfate, 0.0082 to 0.0090g of 7-water zinc sulfate, 0.0062 to 0.0066g of boric acid, 0.00080 to 0.00086g of potassium iodide, 0.00023 to 0.00027g of sodium molybdate, 0.000023 to 0.000027g of copper sulfate, 0.000023 to 0.000027g of cobalt chloride, 0.0276 to 0.0280g of 7-water ferric sulfate, 0.0371 to 0.0375g of disodium ethylenediaminetetraacetate, 0.0018 to 0.0022g of glycine, 0.0003 to 0.0005g of thiamine hydrochloride, 0.0004 to 0.0006g of pyridoxine hydrochloride, 0.08 to 0.08g of inositol, 0.0004 to 0.0004g of indole acetate, 0.0006g to 0.5 g of xanthan gum, and 0.0005 to 5g of benzyl amino acid.
2. The improved MS culture medium of claim 1, wherein the improved MS culture medium comprises the following components in percentage by mass in each liter by taking water as a solvent:
1.31-1.33 g of ammonium sulfate, 1.9-2.1 g of potassium nitrate, 0.36-0.38 g of 7-water magnesium sulfate, 0.165-0.175 g of monopotassium phosphate, 0.43-0.45 g of calcium chloride, 0.0168-0.0170 g of manganese sulfate, 0.0084-0.0088 g of 7-water zinc sulfate, 0.0063-0.0065 g of boric acid, 0.00082-0.00084 g of potassium iodide, 0.00024-0.00026 g of sodium molybdate, 0.000024-0.000026 g of copper sulfate, 0.000024-0.000026 g of cobalt chloride, 0.0277-0.0279 g of 7-water ferric sulfate, 0.0372-0.0374 g of disodium ethylenediaminetetraacetate, 0.0019-0.0021 g of glycine, 0.00035-0.00045 g of thiamine hydrochloride, 0.00045-0.00055 g of pyridoxine hydrochloride, 0.09-0.11 g of inositol, 0.45-0.0006.0006 g of indoleacetic acid, 0.0006-0.00035 g of carrageenan, and 2.0008-2.8 g of benzyl amino acid.
3. The modified MS culture medium of claim 2, wherein the modified MS culture medium comprises the following components in mass per liter in water as a solvent:
1.32g of ammonium sulfate, 2g of potassium nitrate, 0.37g of 7-water magnesium sulfate, 0.17g of monopotassium phosphate, 0.44g of calcium chloride, 0.0169g of manganese sulfate, 0.0086g of 7-water zinc sulfate, 0.0064g of boric acid, 0.00083g of potassium iodide, 0.00025g of sodium molybdate, 0.000025g of copper sulfate, 0.000025g of cobalt chloride, 0.0278g of 7-water ferric sulfate, 0.0373g of disodium ethylenediamine tetraacetate, 0.0020g of glycine, 0.0004g of thiamine hydrochloride, 0.0005g of pyridoxine hydrochloride, 0.1g of inositol, 0.0005g of indoleacetic acid, 0.001g of 6-benzylaminopurine, 4g of carrageenan and 2g of xanthan gum.
4. The modified MS medium of any one of claims 1 to 3, wherein the modified MS medium has a pH of 5.8 to 6.0.
5. The method of any one of claims 1 to 4 for the preparation of an improved MS medium, comprising the steps of:
(1) dissolving components except carrageenan and xanthan gum in water to obtain a liquid MS culture medium;
(2) adding carrageenan and xanthan gum into the liquid MS culture medium, and sterilizing for 15-20 min under the conditions of 0.1-0.15 MPa and 118-121 ℃ to obtain the improved MS culture medium.
6. The use of the modified MS medium of any one of claims 1 to 4 in sweet potato tissue culture.
7. The use of claim 6, wherein the sweetpotato tissue is a sweetpotato shoot apical meristem.
8. The use according to claim 7, wherein the tissue culture method is: inoculating the shoot apical meristem of the sweet potato into the modified MS culture medium for culture.
9. The use of claim 8, wherein the size of the shoot apical meristem of sweetpotato is 0.1 to 0.3 mm.
10. The use according to claim 9, wherein the temperature of the culture is 26-28 ℃, the illumination intensity of the culture is 2800-3200 Lux, the illumination time per day is 12-14 h, and the culture time is 18-22 d.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115968789A (en) * | 2023-02-24 | 2023-04-18 | 福建省农业科学院亚热带农业研究所（福建省农业科学院蔗麻研究中心） | Method for sterilizing and sterile sowing culture of anoectochilus formosanus seeds |
| CN116034875A (en) * | 2023-01-10 | 2023-05-02 | 重庆市铜梁区果之王园艺研究院 | Cherry stock tissue culture seedling culture medium and tissue culture seedling culture method thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5320961A (en) * | 1992-11-16 | 1994-06-14 | Board Of Trustees Operating Michigan State University | Method for asexual in vitro propagation of fertile corn plants |
| CN101138324A (en) * | 2007-09-20 | 2008-03-12 | 江苏徐淮地区徐州农业科学研究所 | Culture medium for inducing clumping bud of sweet potato |
| CN101611697A (en) * | 2009-07-13 | 2009-12-30 | 周玉玲 | Sweet potato merchant 19 detoxifying fast breeding technique and cultivation material |
| CN101831399A (en) * | 2010-05-18 | 2010-09-15 | 华南师范大学 | Method for preparing sweet potato solid culture medium |
| CN103843660A (en) * | 2012-12-07 | 2014-06-11 | 青岛宝依特生物制药有限公司 | Culture medium for inducing sweet potato cluster buds |
| CN110199881A (en) * | 2019-07-10 | 2019-09-06 | 江苏徐淮地区徐州农业科学研究所(江苏徐州甘薯研究中心) | A kind of sweet potato health seedling cultural method removing sweet potato viruses complex SPVD |
-
2022
- 2022-02-09 CN CN202210121567.9A patent/CN114431143B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5320961A (en) * | 1992-11-16 | 1994-06-14 | Board Of Trustees Operating Michigan State University | Method for asexual in vitro propagation of fertile corn plants |
| CN101138324A (en) * | 2007-09-20 | 2008-03-12 | 江苏徐淮地区徐州农业科学研究所 | Culture medium for inducing clumping bud of sweet potato |
| CN101611697A (en) * | 2009-07-13 | 2009-12-30 | 周玉玲 | Sweet potato merchant 19 detoxifying fast breeding technique and cultivation material |
| CN101831399A (en) * | 2010-05-18 | 2010-09-15 | 华南师范大学 | Method for preparing sweet potato solid culture medium |
| CN103843660A (en) * | 2012-12-07 | 2014-06-11 | 青岛宝依特生物制药有限公司 | Culture medium for inducing sweet potato cluster buds |
| CN110199881A (en) * | 2019-07-10 | 2019-09-06 | 江苏徐淮地区徐州农业科学研究所(江苏徐州甘薯研究中心) | A kind of sweet potato health seedling cultural method removing sweet potato viruses complex SPVD |
Non-Patent Citations (21)
| Title |
|---|
| 储凤丽等: "不同培养基对商薯11茎尖分生组织成苗的影响", 《农业科技通讯》 * |
| 储凤丽等: "不同培养基对商薯11茎尖分生组织成苗的影响", 《农业科技通讯》, no. 07, 17 July 2017 (2017-07-17) * |
| 刘扬等: "海南叶用甘薯茎尖脱毒再生技术体系研究", 《热带农业科学》 * |
| 刘扬等: "海南叶用甘薯茎尖脱毒再生技术体系研究", 《热带农业科学》, no. 06, 25 June 2020 (2020-06-25) * |
| 吴翠荣等: "广西甘薯品种的茎尖诱导培养研究初报", 《西南农业学报》 * |
| 吴翠荣等: "广西甘薯品种的茎尖诱导培养研究初报", 《西南农业学报》, no. 01, 28 February 2013 (2013-02-28) * |
| 唐丽雪等: "不同浓度NAA/6BA配比对不同品种甘薯茎尖培养特性的影响", 《中国农学通报》 * |
| 唐丽雪等: "不同浓度NAA/6BA配比对不同品种甘薯茎尖培养特性的影响", 《中国农学通报》, no. 21, 23 July 2020 (2020-07-23) * |
| 唐君等: "外源激素对甘薯不同品种茎尖培养与植株再生的影响", 《江苏农业研究》 * |
| 唐君等: "外源激素对甘薯不同品种茎尖培养与植株再生的影响", 《江苏农业研究》, no. 03, 30 September 2001 (2001-09-30) * |
| 孔祥生等: "生长素类物质对甘薯茎尖分生组织培养成苗的影响", 《山地农业生物学报》 * |
| 孔祥生等: "生长素类物质对甘薯茎尖分生组织培养成苗的影响", 《山地农业生物学报》, no. 02, 28 April 2003 (2003-04-28) * |
| 李晓青等: "甘薯商薯19茎尖脱毒技术研究", 《农业科技通讯》 * |
| 李晓青等: "甘薯商薯19茎尖脱毒技术研究", 《农业科技通讯》, no. 12, 17 December 2012 (2012-12-17), pages 67 - 69 * |
| 林丛发等: "宁薯10号茎尖离体培养及种薯生产", 《广东农业科学》 * |
| 林丛发等: "宁薯10号茎尖离体培养及种薯生产", 《广东农业科学》, no. 10, 25 May 2013 (2013-05-25) * |
| 王颖: "甘薯高产栽培茎尖组培快繁方法", 《乡村科技》 * |
| 王颖: "甘薯高产栽培茎尖组培快繁方法", 《乡村科技》, no. 13, 10 May 2018 (2018-05-10) * |
| 翟红莲 等: ""植物组织培养培养基凝固剂替代物的研究"", no. 2, pages 5 - 9 * |
| 阮先乐等: "甘薯茎尖分生组织培养及快繁技术的研究", 《周口师范学院学报》 * |
| 阮先乐等: "甘薯茎尖分生组织培养及快繁技术的研究", 《周口师范学院学报》, no. 02, 15 March 2015 (2015-03-15) * |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116034875A (en) * | 2023-01-10 | 2023-05-02 | 重庆市铜梁区果之王园艺研究院 | Cherry stock tissue culture seedling culture medium and tissue culture seedling culture method thereof |
| CN115968789A (en) * | 2023-02-24 | 2023-04-18 | 福建省农业科学院亚热带农业研究所（福建省农业科学院蔗麻研究中心） | Method for sterilizing and sterile sowing culture of anoectochilus formosanus seeds |
| CN115968789B (en) * | 2023-02-24 | 2024-02-13 | 福建省农业科学院亚热带农业研究所（福建省农业科学院蔗麻研究中心） | Anoectochilus roxburghii seed disinfection and aseptic seeding culture method |
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