Garlic cultivation MS culture medium formula

Abstract

The invention discloses a garlic cultivation MS culture medium formula which is prepared from the following raw materials in parts by mass: 1-3 parts of garlic extract, 12-18 parts of agar, 8-10 parts of a carbon source, 3-5 parts of yeast extract, 10-15 parts of powdered calcium, 3-5 parts of trace elements, 10-12 parts of inorganic salt, 5-8 parts of a nitrogen source, 0.5-1 part of plant growth hormone, 1-1.5 parts of a water-retaining agent, 0.5-1.5 parts of a stabilizer and 20-30 parts of distilled water. The water-retaining agent is added in the culture medium, the water-retaining agent is arranged, so that garlic culture tissues are in the culture medium, and the water-retaining agent can guarantee the humidity during tissue germination, so that the culture tissues can better absorb nutrition for seedling culture; the stability of each element component in the culture medium is further guaranteed, the cleanliness of the culture medium is guaranteed, the culture medium is prevented from being infected by microorganisms, and the stable absorption of nutrient components by cultured tissues is guaranteed.

Description

Technical Field

[0001] The invention relates to the technical field of garlic tissue culture, in particular to a garlic cultivation MS culture medium formula. Background Art

[0002] MS medium is a plant culture medium, commonly used in plant tissue culture and plant biology research. MS medium contains inorganic salts, organic matter, vitamins and plant hormones, and can provide all the nutrients required for plant growth.

[0003] The existing garlic culture is usually induced into seedlings by culturing garlic stem tips, root tips, flower tips, leaves and garlic stem tip meristems in a culture medium, and then transplanted to the field or induced into test tubes for further seedling selection and seedling raising. The components of the existing garlic culture medium are mostly composed of inorganic salts, organic matter, trace elements, carbon sources and plant hormones. Although this type of culture medium can provide the nutrients and trace elements required for garlic culture, in the actual culture process, due to the different types of tissues cultured in different garlics and the different activities of the selected cultured tissues, these cultured tissues absorb the nutrients in the culture medium differently, and one of the properties that affects the absorption is water retention. Therefore, when cultivating garlic, the existing culture medium cannot well guide the cultured tissues to absorb the nutrients in the culture medium. Summary of the invention

[0004] The object of the present invention is to provide a garlic cultivation MS medium formula to solve the problems raised in the above background technology.

[0005] To achieve the above object, the present invention provides the following technical scheme: a garlic cultivation MS culture medium formula, which is composed of the following raw materials in proportion by mass, including 1-3 parts of garlic extract, 12-18 parts of agar, 8-10 parts of carbon source, 3-5 parts of yeast extract, 10-15 parts of powdered calcium, 3-5 parts of trace elements, 10-12 parts of inorganic salts, 5-8 parts of nitrogen source, 0.5-1 part of plant growth hormone, 1-1.5 parts of water retaining agent, 0.5-1.5 parts of stabilizer and 20-30 parts of distilled water.

[0006] Preferably, the carbon source is prepared from the following raw materials according to a mass ratio, including 75%-80% sucrose, 10%-15% glucose and 5%-8% fructose.

[0007] Preferably, the trace elements are prepared from the following raw materials according to mass ratio, including 10%-15% boric acid, 20%-25% manganese sulfate, 30%-45% zinc sulfate, 3%-8% sodium molybdate, 3%-5% iron sulfate, 1%-3% potassium iodide and 1%-2% cobalt chloride.

[0008] Preferably, the yeast extract includes yeast extract, yeast amino acid salt and yeast extract.

[0009] Preferably, the inorganic salt is prepared from the following raw materials according to mass ratio, including 15%-25% ammonium nitrate, 35%-45% potassium nitrate, 10%-15% potassium dihydrogen phosphate, 10%-15% magnesium sulfate and 10%-15% calcium chloride.

[0010] Preferably, the nitrogen source is prepared from the following raw materials according to mass ratio, including 30%-35% amino acid, 10%-20% calcium nitrate, 25%-30% ammonium chloride, 8%-10% nitroamide and 10%-15% ammonia nitrogen compound.

[0011] Preferably, the amino acid is composed of the following raw materials in proportion by mass, including 8-10 parts of aspartic acid, 5-8 parts of glutamic acid and 3-5 parts of lysine.

[0012] Preferably, the plant growth hormone is composed of the following raw materials in a mass fraction ratio, including 3-5 parts of indoleacetic acid and 1-3 parts of cytokinin.

[0013] Preferably, the water retaining agent comprises one or more of disintegrating rock, sodium succinate, polyacrylamide and gelatin enzymatic hydrolysate.

[0014] Preferably, the stabilizer comprises one or more of ascorbic acid, sodium thiosulfate, phosphate buffer and HEPES buffer.

[0015] Technical effects and advantages of the present invention:

[0016] The present invention adds a water retaining agent to the culture medium. Through the setting of the water retaining agent, the garlic culture tissue is in the culture medium, and the water retaining agent can ensure the humidity when the tissue germinates, thereby ensuring that the culture tissue can better absorb nutrients for seedling cultivation. At the same time, a stabilizer is set in the culture medium formula. Through the stable setting, the stability of each element component in the culture medium is further ensured, the cleanliness of the culture medium is ensured, the culture medium is prevented from being infected by microorganisms, and the culture tissue is ensured to stably absorb nutrients. DETAILED DESCRIPTION

[0017] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0018] The invention provides a garlic cultivation MS culture medium formula, which is composed of the following raw materials in proportion by mass, including 1-3 parts of garlic extract, 12-18 parts of agar, 8-10 parts of carbon source, 3-5 parts of yeast extract, 10-15 parts of powdered calcium, 3-5 parts of trace elements, 10-12 parts of inorganic salts, 5-8 parts of nitrogen source, 0.5-1 part of plant growth hormone, 1-1.5 parts of water retaining agent, 0.5-1.5 parts of stabilizer and 20-30 parts of distilled water.

[0019] It should be noted that the following steps are included in the preparation of the culture medium:

[0020] The first step is to add garlic extract, carbon source, yeast extract, powdered calcium, etc. into distilled water and stir evenly to obtain a primary mixed solution;

[0021] The second step is to add trace elements, inorganic salts, nitrogen sources and plant growth hormones to the primary mixed solution obtained in the first step, and stir and mix them evenly to obtain a mixed stock solution;

[0022] The third step is to add a water retaining agent and a stabilizer to the mixed stock solution obtained in the second step, and stir and mix to obtain a culture solution;

[0023] Step 4, adding agar to the culture solution prepared in step 3, stirring evenly to obtain a culture medium;

[0024] Step 5. Heat the culture medium to boil for one minute, then accelerate cooling to 45-50°C, pour it into a culture dish, lay it flat, and heat it to 37°C in an incubator after it solidifies.

[0025] Agar is a commonly used gelling agent, also known as agarose or gelatin, which is usually extracted from seaweed or animal bones. Agar has good gel properties and can form a solid gel within a certain temperature range, making it one of the ideal choices for making agar culture medium. In the culture medium, agar can provide a solid matrix to support the growth of plants or microorganisms without causing harmful effects on them. At the same time, agar can also help the culture medium maintain a certain degree of moisture and promote the growth of plants or microorganisms.

[0026] Powdered calcium is a common calcium supplement, also known as calcium carbonate. In plant cultivation and agriculture, powdered calcium is often used to adjust soil pH and provide calcium required by plants. Calcium is an essential element for plant growth and development, and it participates in physiological processes such as cell wall construction, cell division and cytoplasmic transport. Powdered calcium can be quickly dissolved in the soil to release calcium ions that can be absorbed and utilized by plants, thereby improving soil texture, promoting plant growth and increasing crop yields. In plant culture media, powdered calcium can also be used to adjust the pH value of the culture medium and provide the necessary calcium element, providing the necessary nutritional support for the normal growth of plants.

[0027] Specifically, the carbon source is prepared from the following raw materials according to mass ratio, including 75%-80% sucrose, 10%-15% glucose and 5%-8% fructose.

[0028] It should be noted that the carbon source in the culture medium is a compound or substance that provides the carbon element required for the growth and metabolism of microorganisms or plants. Glucose is a simple carbon source that microorganisms and plants can quickly use for energy and growth. Sucrose is also a commonly used carbon source that can provide a stable carbon source under certain culture conditions.

[0029] Specifically, the trace elements are prepared from the following raw materials according to mass ratio, including 10%-15% boric acid, 20%-25% manganese sulfate, 30%-45% zinc sulfate, 3%-8% sodium molybdate, 3%-5% iron sulfate, 1%-3% potassium iodide and 1%-2% cobalt chloride.

[0030] It should be noted that trace elements are added to the culture medium to meet the growth needs of plants or microorganisms. Among them, boric acid promotes plant growth, cell division and fruit development; manganese sulfate participates in photosynthesis and maintains the synthesis of chlorophyll; zinc sulfate promotes plant growth and regulates material metabolism; sodium molybdate participates in nitrogen metabolism and nitrogen fixation; iron sulfate participates in plant respiration and photosynthesis; potassium iodide promotes plant growth and photosynthesis; cobalt chloride promotes plant growth and is an essential element for the synthesis of vitamin B12.

[0031] Specifically, yeast extracts include yeast extract, yeast amino acid salts and yeast extract.

[0032] It should be noted that yeast extract is rich in amino acids, vitamins and other growth factors, which promote plant growth and development. Yeast extract is usually added to the culture medium to improve the nutritional value and growth effect of the culture medium. In the process of cultivating plants, the appropriate addition of yeast extract can promote plant growth, root development and physiological activity, and contribute to the healthy growth of plants.

[0033] Yeast extract is a mixture extracted from yeast cells, which contains a variety of nutrients such as amino acids, nucleic acids, vitamins and enzymes. These substances can provide plants with the nutrients they need and promote their growth and development. Adding an appropriate amount of yeast extract to the culture medium can improve the nutritional value of the culture medium, enhance the growth ability of plants, and promote the root growth and physiological activity of plants.

[0034] Yeast amino acid salt is a salt mixture rich in amino acids, usually extracted from yeast cells. These amino acids are one of the important nutrients required by plants and play an important role in the growth and development of plants. Adding an appropriate amount of yeast amino acid salt to the culture medium can increase the amino acid content of the culture medium, provide sufficient nitrogen and carbon sources, and thus promote the growth and development of plants. In addition, yeast amino acid salt also contains other bioactive substances, such as growth factors, which can further promote the root growth and nutrient absorption of plants. Therefore, yeast amino acid salt is also a commonly used plant culture medium additive, which helps to improve the quality of the culture medium and the growth effect of plants.

[0035] Yeast extract is a nutrient extract derived from yeast cells, rich in protein, amino acids, vitamins and minerals. Yeast extract is widely used in microbiology, cell culture and biotechnology. It can be used as part of cell culture medium to provide nutrients required for cell growth and differentiation, promote cell growth, proliferation and expression of target proteins, etc. Yeast extract is also commonly used to cultivate fungi and yeast strains to promote their growth and reproduction.

[0036] Specifically, the inorganic salt is prepared from the following raw materials according to mass ratio, including 15%-25% ammonium nitrate, 35%-45% potassium nitrate, 10%-15% potassium dihydrogen phosphate, 10%-15% magnesium sulfate and 10%-15% calcium chloride.

[0037] It should be noted that inorganic salts are used to provide the main nutrients needed by plants.

[0038] Specifically, the nitrogen source is prepared from the following raw materials according to mass ratio, including 30%-35% amino acid, 10%-20% calcium nitrate, 25%-30% ammonium chloride, 8%-10% nitroamide and 10%-15% ammonia nitrogen compound, and is used to promote plant growth.

[0039] Furthermore, the amino acid is composed of the following raw materials in proportion by mass, including 8-10 parts of aspartic acid, 5-8 parts of glutamic acid and 3-5 parts of lysine.

[0040] It should be noted that adding aspartic acid, glutamic acid and lysine to plant culture medium can provide the amino acid nutrition required by plants. These amino acids play an important role in plant growth and development, and are involved in physiological activities such as protein synthesis and metabolic regulation. By adding an appropriate amount of amino acids to the culture medium, the growth, proliferation and differentiation of plant tissues can be promoted, and the culture effect and quality can be improved. At the same time, amino acids can also enhance the resistance of plants to adverse stress and help improve the growth environment and growth conditions of plants. In plant tissue culture and related research, the addition of aspartic acid, glutamic acid and lysine can play an important role in nutritional support.

[0041] Specifically, the plant growth hormone is composed of the following raw materials in a mass fraction ratio, including 3-5 parts of indoleacetic acid and 1-3 parts of cytokinin.

[0042] It should be noted that indoleacetic acid is a plant growth hormone that mainly regulates plant cell division, cell elongation and root morphological development. Cytokinin promotes cell division and differentiation, and promotes plant growth and development.

[0043] Specifically, the water retaining agent includes one or more of disintegrating rock, sodium succinate, polyacrylamide and gelatin enzymatic hydrolysate.

[0044] It should be noted that disintegrating rock is a natural mineral with good water retention properties. It can effectively absorb and retain water, and is also beneficial to the aeration of the culture medium. Sodium succinate can absorb a large amount of water and form a gel, keeping the culture medium moist while providing a certain support effect, similar to agar. Polyacrylamide has good water absorption properties and can absorb water on its surface to form a hydrogel, maintaining the water balance of the culture medium. Gelatin enzymatic hydrolysate can also be used as a water retaining agent, with water absorption properties and moisturizing effects.

[0045] Specifically, the stabilizer includes one or more of ascorbic acid, sodium thiosulfate, phosphate buffer and HEPES buffer.

[0046] It should be noted that ascorbic acid and sodium thiosulfate are antioxidants that can protect some easily oxidized components in the culture medium and prevent oxidation reactions. Phosphate buffer and HEPES buffer are pH buffers that are used to adjust the pH value of the culture medium and maintain the stability of the acidity and alkalinity of the culture medium.

[0047] In addition to the above stabilizers, antimicrobial agents such as antibiotics and antifungal agents can be added to effectively inhibit microbial contamination in the culture medium and keep the culture medium clean.

[0048] Finally, it should be noted that the above is only a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the aforementioned embodiments, it is still possible for those skilled in the art to modify the technical solutions described in the aforementioned embodiments or to make equivalent substitutions for some of the technical features therein. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A garlic cultivation MS medium formula, characterized in that: The invention is composed of the following raw materials in proportion by mass, including 1-3 parts of garlic extract, 12-18 parts of agar, 8-10 parts of carbon source, 3-5 parts of yeast extract, 10-15 parts of powdered calcium, 3-5 parts of trace elements, 10-12 parts of inorganic salts, 5-8 parts of nitrogen source, 0.5-1 part of plant growth hormone, 1-1.5 parts of water retaining agent, 0.5-1.5 parts of stabilizer and 20-30 parts of distilled water.

2. A garlic cultivation MS medium formula according to claim 1, characterized in that, The carbon source is prepared from the following raw materials according to mass ratio, including 75%-80% sucrose, 10%-15% glucose and 5%-8% fructose.

3. A garlic cultivation MS medium formula according to claim 1, characterized in that, The trace elements are prepared from the following raw materials according to mass ratio, including 10%-15% boric acid, 20%-25% manganese sulfate, 30%-45% zinc sulfate, 3%-8% sodium molybdate, 3%-5% iron sulfate, 1%-3% potassium iodide and 1%-2% cobalt chloride.

4. A garlic cultivation MS medium formula according to claim 1, characterized in that, The yeast extract includes yeast extract, yeast amino acid salt and yeast extract.

5. A garlic cultivation MS medium formula according to claim 1, characterized in that: The inorganic salt is prepared from the following raw materials according to mass ratio, including 15%-25% of ammonium nitrate, 35%-45% of potassium nitrate, 10%-15% of potassium dihydrogen phosphate, 10%-15% of magnesium sulfate and 10%-15% of calcium chloride.

6. A garlic cultivation MS medium formula according to claim 1, characterized in that: The nitrogen source is prepared from the following raw materials according to mass ratio, including 30%-35% of amino acid, 10%-20% of calcium nitrate, 25%-30% of ammonium chloride, 8%-10% of nitramide and 10%-15% of ammonia nitrogen compound.

7. A garlic cultivation MS medium formula according to claim 6, characterized in that: The amino acid is composed of the following raw materials in proportion by mass, including 8-10 parts of aspartic acid, 5-8 parts of glutamic acid and 3-5 parts of lysine.

8. A garlic cultivation MS medium formula according to claim 1, characterized in that: The plant growth hormone is composed of the following raw materials in a mass fraction ratio, including 3-5 parts of indoleacetic acid and 1-3 parts of cytokinin.

9. A garlic cultivation MS medium formula according to claim 1, characterized in that: The water retaining agent comprises one or more of disintegrating rock, sodium succinate, polyacrylamide and gelatin enzymatic hydrolysate.

10. The MS medium formula for garlic cultivation according to claim 1, characterized in that: The stabilizer includes one or more of ascorbic acid, sodium thiosulfate, phosphate buffer and HEPES buffer.