CN115843686A - Culture medium for sweet potato tissue culture and preparation method thereof - Google Patents
Culture medium for sweet potato tissue culture and preparation method thereof Download PDFInfo
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- CN115843686A CN115843686A CN202211544066.8A CN202211544066A CN115843686A CN 115843686 A CN115843686 A CN 115843686A CN 202211544066 A CN202211544066 A CN 202211544066A CN 115843686 A CN115843686 A CN 115843686A
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Abstract
The invention discloses a culture medium for sweet potato tissue culture and a preparation method thereof. The invention changes calcium chloride of the original culture medium into calcium nitrate, changes ammonium nitrate into ammonium chloride, and simultaneously properly adds urea and potassium hydroxide to increase the content of nitrogen source and potassium element, thereby meeting the requirements of potassium element and nitrogen element of potassium-like crops such as sweet potatoes and ensuring that the micro-ecosystem ions in the tissue culture bottle are in a balanced state beneficial to the growth of the sweet potatoes. The method effectively avoids using ammonium nitrate when the culture medium is prepared, avoids risks caused by storing the ammonium nitrate, and can also avoid the influence on the tissue culture of the sweet potatoes after the ammonium nitrate is forbidden in China.
Description
Technical Field
The invention belongs to the technical field of agriculture, and particularly relates to a culture medium for sweet potato tissue culture and a preparation method thereof.
Background
The commonly used culture medium for the sweet potato group is MS culture medium. Wherein, ammonium nitrate is one of the important components of the MS culture medium, and the main function of the ammonium nitrate is to provide a nitrogen source (ammonium nitrogen) for the plant tissue culture medium. However, ammonium nitrate is unstable in nature and can be decomposed by explosion when being subjected to strong impact or heat, so that the ammonium nitrate belongs to dangerous goods ranks and has higher requirements on production, storage and transportation conditions.
In addition, the production of ammonium nitrate is forbidden at present in China, so that the existing MS culture gene is lack of ammonium nitrate and cannot be normally prepared and used, and the tissue culture production of sweet potatoes is influenced.
Therefore, a culture medium which does not use ammonium nitrate and has a good tissue culture effect is required.
Disclosure of Invention
The invention aims to overcome the existing problems and provides a culture medium for sweet potato tissue culture and a preparation method thereof. In order to achieve the purpose, the invention adopts the following technical scheme:
the invention discloses a culture medium for sweet potato tissue culture, which comprises the following components in parts by weight:
as an improvement, the raw materials of the culture medium comprise the following components in percentage by weight:
the invention also discloses a preparation method of the culture medium for sweet potato tissue culture, which comprises the following steps S1, preparing a mother solution or dry powder culture medium;
s2, taking mother liquor according to a proportion, and sequentially dissolving the mother liquor in deionized water;
s3, adding sucrose and sweet potato auxin as required;
s4, fixing the volume;
s5, adjusting the PH value;
s6, filling into a culture bottle;
s7, sterilizing processing;
and S8, cooling to normal temperature.
In the improvement, when the mother liquor is prepared in the step S1, the mother liquor is prepared according to reagent number groups respectively, and then the mother liquor multiple and the using amount are configured.
As an improvement, when mother liquor is prepared in groups in the step S1, the medicine components in each group of reagents are completely dissolved in sequence.
In the modification, after the pH is adjusted in the step S5, agar and carrageenan are added according to the use condition.
As an improvement, the temperature of the sterilization treatment in the step S7 is 121 ℃, and the sterilization time is 20min.
The invention has the advantages that:
1. the invention changes calcium chloride of the original culture medium into calcium nitrate, changes ammonium nitrate into ammonium chloride, and simultaneously properly adds urea and potassium hydroxide to increase the content of nitrogen source and potassium element, thereby meeting the requirements of potassium element and nitrogen element of potassium-like crops such as sweet potatoes and ensuring that the micro-ecosystem ions in the tissue culture bottle are in a balanced state beneficial to the growth of the sweet potatoes.
2. The method effectively avoids using ammonium nitrate when the culture medium is prepared, avoids risks caused by storing the ammonium nitrate, and can also avoid the influence on the tissue culture of the sweet potatoes after the ammonium nitrate is forbidden in China.
Drawings
FIG. 1 is a graph showing the effect of the test group after 40 days of culture as compared with the control group.
FIG. 2 is a graph showing the data of samples after 30 days of culture.
FIG. 3 is a graph of sampling data after 40 days of culture.
Detailed Description
The present invention will be described in detail and specifically with reference to the following examples so as to facilitate the understanding of the present invention, but the following examples do not limit the scope of the present invention.
Example 1
The embodiment discloses a culture medium for sweet potato tissue culture, which comprises the following components in parts by weight:
the preparation method of the embodiment comprises the following steps:
s1, preparing mother liquor, and preparing mother liquor multiples and using amount according to actual needs:
s1-1, preparation of mother liquor of group A1 reagent
Weighing 50 times of the weight of the magnesium sulfate heptahydrate and the potassium dihydrogen phosphate in the A1 group of reagents by using an electronic balance, sequentially dissolving the two components in 900ml of deionized water (one reagent needs to be dissolved completely first, and the other reagent needs to be dissolved later), and after all the reagents are completely dissolved, fixing the volume to 1L and marking the volume as 'A1 mother liquor' for later use. Example (c): the content of magnesium sulfate heptahydrate per liter of culture medium is as follows: 370mg, 50 times its weight is: 18500mg, i.e. mother liquor content 18.5g/L, the rest reagents are calculated according to this category.
S1-2, mother liquor for preparing group A2 reagent
Weighing 50 times of the components of the group A2 reagents by using an electronic balance, dissolving the components in 900ml of deionized water (one reagent needs to be dissolved completely first and the other needs to be dissolved completely), and then fixing the volume to 1L of the labeled "A2 mother liquor" for later use.
S1-3, preparing mother liquor of reagent in group B1
Weighing the boric acid, manganese sulfate monohydrate, zinc sulfate, potassium iodide, sodium molybdate, copper sulfate pentahydrate and cobalt chloride hexahydrate in the reagent B1 by using an electronic balance to obtain 100 times of the weight of the components, sequentially dissolving the components in 900ml of deionized water (one reagent needs to be dissolved completely, and the other reagent needs to be dissolved completely), and then fixing the volume until 1L of the reagent is marked with 'B1 mother liquor' for later use.
S1-4, mother liquor for preparing C1 group reagent
Weighing two components of ferrous sulfate and disodium ethylene diamine tetraacetate in the C1 group of reagents by using an electronic balance and the like, weighing 50 times of the weight of the components, sequentially dissolving the components in 900ml of deionized water (one reagent needs to be completely dissolved first, and the other needs to be dissolved later), and after all the reagents are completely dissolved, fixing the volume to 1L and marking a 'C1 mother solution' for later use.
S1-5 mother liquor for preparing group D1 reagents
The five components of inositol, nicotinic acid VB3, pyridoxine hydrochloride VB6, thiamine hydrochloride VB1 and glycine in the group D1 reagent are weighed by using an electronic balance and the like, the weight of the five components is 100 times of the weight of the five components, the five components are sequentially dissolved in 900ml of deionized water (one reagent needs to be dissolved completely and the other needs to be dissolved completely), and after all the reagents are dissolved completely, the volume is determined to 1L of labeled 'D1 mother liquor' for standby.
The mother liquor prepared above needs to be stored in a refrigerator refrigerating chamber under the condition of not using. Any method may be used as long as no crystallization occurs.
S2, taking mother liquor in proportion, and dissolving the mother liquor in deionized water in sequence
Taking the mother liquor according to the proportion, wherein the taking amount is as follows: a1:20mL, A2. And then dissolved in 800mL of deionized water in sequence.
S3, adding 30g/L of cane sugar, sweet potato auxin and the like according to requirements.
And S4, fixing the volume of the dissolved solution to 1L.
S5, adjusting the pH value of the culture medium to a required value; then agar and carrageenan are put or not put according to the use condition. In this example, 7g of agar was selected.
And S6, filling into a culture bottle.
S7, sterilizing for 20 minutes by using a sterilizing pot at 121 ℃.
S8, cooling the culture medium and then using the culture medium.
Example 2
The raw material components of this example are:
the manufacturing method of the embodiment comprises the following steps:
s1, preparing a dry powder culture medium by a drying method after reagents are proportioned,
s2, weighing 4.74g according to the process requirements, and dissolving in 1L of deionized water.
S3, adding 30g/L of cane sugar, sweet potato auxin and the like according to requirements.
And S4, fixing the volume of the dissolved solution to 1L.
S5, adjusting the pH value of the culture medium to a required value; then agar and carrageenan are put or not put according to the use condition. In this example, 7g of agar was selected.
And S6, filling into a culture bottle.
S7, sterilizing for 20 minutes by using a sterilizing pot at 121 ℃.
S8, cooling the culture medium and then using the culture medium.
Example 3
In this example, the raw materials and the mixture ratio of the culture medium are as follows:
the manufacturing method of this embodiment is the same as embodiment 1.
Example 4
In this example, the raw materials and the mixture ratio of the culture medium are as follows:
the manufacturing method of this example is the same as example 1.
The invention sets a contrast test to verify the culture effect of the culture medium:
(1) Test method
The method comprises the steps of taking Pushu No. 32 virus-free seedlings, respectively placing the seedlings in culture media (shown in table 1) of different treatments, culturing under the illumination of 13h/24h at 28 ℃, keeping 1-2 axillary buds with the length of each stem section being about 1cm, removing leaf stalks, culturing 4 seedlings in each bottle, treating 30 bottles in each bottle, and measuring the root number, stem base diameter, plant height and leaf number of each tissue culture seedling at 30 days and 40 days. The initial time of the test was 2021, 10 months and 18 days.
Table 1 respective test protocols
(2) Test results
Data were collected on day 30 on day 11/17 of 2021 for each test group and control group. The results are as follows:
TABLE 2 basic data of each tissue culture seedling at 30 days
On day 27/11/2021, data were collected on day 30 for each test group and control group. The results are as follows:
TABLE 3 basic data of each tissue culture seedling at 40 days
Test results show that the culture medium still has good culture effect after calcium chloride is replaced by calcium nitrate instead of ammonium nitrate. Among them, the growth effect of the cultured seedlings in SMP04 (example 3) was the best and superior to that of the cultured seedlings in the medium using ammonium nitrate.
The embodiments of the present invention have been described in detail above, but they are merely exemplary, and the present invention is not equivalent to the embodiments described above. Any equivalent modifications and substitutions to those skilled in the art are also within the scope of the present invention. Accordingly, it is intended that all equivalent alterations and modifications be included within the scope of the invention, without departing from the spirit and scope of the invention.
Claims (7)
1. The culture medium for sweet potato tissue culture is characterized by comprising the following components in parts by weight:
2. the culture medium for sweet potato tissue culture according to claim 1, wherein the sweet potato group culture medium comprises the following components in parts by weight:
3. the method for preparing the culture medium for sweet potato tissue culture according to claim 1, which comprises the following steps
S1, preparing a mother solution or dry powder culture medium;
s2, taking mother liquor according to a proportion, and sequentially dissolving the mother liquor in deionized water;
s3, adding sucrose and sweet potato auxin as required;
s4, fixing the volume;
s5, adjusting the PH value;
s6, filling into a culture bottle;
s7, sterilizing;
and S8, cooling to normal temperature.
4. The method for preparing the culture medium for sweet potato tissue culture according to claim 3, wherein the preparation of the mother liquor in the step S1 is carried out by grouping according to reagent numbers, and then the multiple and the amount of the mother liquor are prepared.
5. The method for preparing the culture medium for sweet potato tissue culture according to claim 3, wherein the drug ingredients in each group of reagents are completely dissolved in sequence when the mother solution is prepared in groups in the step S1.
6. The method for preparing a culture medium for sweet potato tissue culture according to claim 3, wherein agar and carrageenan are added according to the use condition after adjusting the pH in the step S5.
7. The method for preparing the culture medium for sweet potato tissue culture according to claim 3, wherein the temperature of the sterilization treatment in the step S7 is 121 ℃, and the sterilization time is 20min.
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| JPH01128784A (en) * | 1987-11-13 | 1989-05-22 | Snow Brand Milk Prod Co Ltd | Production of cell capable of highly producing vitamin k1 by tissue cultivation of ipomoea batatas l. |
| CN101933455A (en) * | 2009-07-03 | 2011-01-05 | 中国科学院上海生命科学研究院 | In vitro propagation method for cinnamomum japonicum |
| US20110035844A1 (en) * | 2006-09-25 | 2011-02-10 | Pioneer Hi-Bred International, Inc. | The Maize ERECTA Genes for Improving Plant Growth, Transpiration, Efficiency and Drought Tolerance in Crop Plants |
| CN102369879A (en) * | 2010-08-10 | 2012-03-14 | 中国科学院植物研究所 | Succeeding preservation method for embryogenic callus of China fir and subculture medium used therein |
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| CN111109083A (en) * | 2020-01-16 | 2020-05-08 | 河南华薯农业科技有限公司 | Culture medium suitable for cultivating multiple sweet potato seedlings and preparation method thereof |
| CN111937745A (en) * | 2020-08-13 | 2020-11-17 | 江苏宝德农业科技有限公司 | Tissue culture method for rapid propagation of tissue culture seedlings for field planting in production process of detoxified miniature potatoes |
| CN114711144A (en) * | 2022-05-05 | 2022-07-08 | 天津博奥聚能生物科技有限公司 | Rapid propagation method and culture medium for dioscorea composita tissue culture seedlings |
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2022
- 2022-12-03 CN CN202211544066.8A patent/CN115843686A/en active Pending
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|---|---|---|---|---|
| JPH01128784A (en) * | 1987-11-13 | 1989-05-22 | Snow Brand Milk Prod Co Ltd | Production of cell capable of highly producing vitamin k1 by tissue cultivation of ipomoea batatas l. |
| US20110035844A1 (en) * | 2006-09-25 | 2011-02-10 | Pioneer Hi-Bred International, Inc. | The Maize ERECTA Genes for Improving Plant Growth, Transpiration, Efficiency and Drought Tolerance in Crop Plants |
| CN101933455A (en) * | 2009-07-03 | 2011-01-05 | 中国科学院上海生命科学研究院 | In vitro propagation method for cinnamomum japonicum |
| CN102369879A (en) * | 2010-08-10 | 2012-03-14 | 中国科学院植物研究所 | Succeeding preservation method for embryogenic callus of China fir and subculture medium used therein |
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| CN111109083A (en) * | 2020-01-16 | 2020-05-08 | 河南华薯农业科技有限公司 | Culture medium suitable for cultivating multiple sweet potato seedlings and preparation method thereof |
| CN111937745A (en) * | 2020-08-13 | 2020-11-17 | 江苏宝德农业科技有限公司 | Tissue culture method for rapid propagation of tissue culture seedlings for field planting in production process of detoxified miniature potatoes |
| CN114711144A (en) * | 2022-05-05 | 2022-07-08 | 天津博奥聚能生物科技有限公司 | Rapid propagation method and culture medium for dioscorea composita tissue culture seedlings |
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