The growth and development of mycelium and fruiting bodies of Ashwagandha cannot be separated from the nutritional conditions, and the nutrients are divided into four major categories, i.e., carbon, nitrogen, inorganic salts (minerals), and growth factors.
(I) Carbon Half of the dry weight of the cell of the gray tree flower is composed of carbon, which shows the importance of the need for carbon in the growth and development of the gray tree flower. Carbon provides two basic functions in the growth and development of the ashwagandha, firstly it provides the required carbon for the synthesis of key cellular components and it forms the basic skeleton of these key components, for example, sugars (carbohydrates), proteins, fats and nucleic acids; and secondly the oxidizing process of the carbon source provides the energy source for the basic life processes of the ashwagandha.
The ashwagandha has no photosynthetic capacity, that is, it can not fix the carbon dioxide in the air, it absorbs and utilizes carbon from the carbonaceous organic matter in the substrate, such as cellulose, hemicellulose, lignin, starch, sucrose, glucose, certain organic acids and certain alcohols. Among the common carbon sources, all small molecule compounds such as monosaccharides, organic acids and alcohols can be directly absorbed by mycelial cells, while large molecule compounds such as cellulose, hemicellulose, lignin and starch can not be directly absorbed, but must be broken down into glucose, arabinose, xylose, galactose and fructose through cellulases, amylases, hemicellulases and ligninases of ashwagandha before they can be absorbed and utilized. Glucose, malt extract, yeast paste, potato juice, woodchip juice, and soluble starch are better carbon sources in the parent species culture. Carbon sources for the original and cultivated species are mainly from chestnut wood chips (or wood chips of close genera and species), cottonseed hulls, sucrose and bran.
1. Glucose is a carbon source that can be directly absorbed and utilized by the mycelium of ashwagandha. Therefore, glucose is often added in the preparation of culture medium. Glucose is a kind of monosaccharide containing aldehyde group, molecular formula for C6H12O6, it is the most important kind of monosaccharide in the organism. It is industrially produced by the hydrolysis of starch, is a white or slightly yellow crystalline powder, soluble in water, and the sweetness is 70% of sucrose. The dosage is 1% to 2% in Ashwagandha parent species culture medium or in woodchip-based culture material. Glucose as a carbon source, ashwagandha growth is the best, mycelium get the largest amount, in line with the physiological metabolic characteristics of the fungus, the main respiratory pathway carbon source of the fungus are phosphorylation of glucose derivatives as its starting point.
Ashwagandha grows better in glucose than in fructose, indicating that aldose is more easily utilized by ashwagandha than ketose. Maltose is a disaccharide consisting of 2 glucose monomers linked together, and sucrose is a disaccharide consisting of glucose and fructose. As a result, the mycelial growth of ashwagandha in the medium with maltose as the carbon source was more than twice as much as on sucrose. Sucrose is usually eaten red and white sugar, by a molecule of glucose and a molecule of fructose condensed into a disaccharide, the molecular formula is C12H22O11. widely found in sugar cane and sugar beets, sweet taste, the crystallization of a large monoclinic crystal shape, very easy to dissolve in water. When heated with acid*** or under the influence of sucrase, sucrose is hydrolyzed to form equal molecules of glucose and fructose, and its mixture is called invert sugar. It is a commonly used component in the preparation of edible mushroom culture medium, and the concentration is generally 1% to 2.5%.
The ability of ashwagandha to utilize monosaccharides and their derivatives varies widely, although some monosaccharides are similar in biochemical properties. Glucose, galactose, and mannose are all six-carbon hexoses, with glucose being a good carbon source that ashwagandha prefers to utilize. Fructose is a five-carbon ketoaldehyde sugar and is second only to glucose as a good carbon source. Other monosaccharides such as sorbose, arabinose, xylose, and rhamnose are monosaccharides that are rarely utilized by ashwagandha. Although certain monosaccharides do not maximize the growth of ashwagandha as much as glucose, it can be seen that the closer the structure of the monosaccharides is to glucose, the more they are utilized by ashwagandha. This may be because the ability of the ashwagandha to utilize a particular monosaccharide depends on how easily it can be converted into a phosphorylated glucose derivative that can enter the respiratory pathway.
Many sugar alcohols, such as sorbitol, mannitol, and glycerol, can be absorbed and utilized as a carbon source by gray trellises, but usually not as effectively as monosaccharides. Mannitol is an exception because it is derived from fructose or mannose reduction, and it can give some ashwagandha the same growth results as on glucose.
Absorption of monosaccharides is accomplished by both assisted diffusion and active transport. The uptake of glucose uses assisted diffusion and active transport, and the uptake of alginate and maltose is active transport. The carrier molecules are proteins and their activity is affected by temperature, pH and inhibitors. Because hydrogen ions intervene in the isotropic transfer of many molecules, pH is particularly important, and active transport is influenced by chemical and physical factors that affect cellular respiration and energy production. In addition, specific inorganic ions, such as potassium ions (K+), affect uptake by acting as cofactors of transfer, and they maintain electron neutrality in proton isotropic transfer, which may affect cellular osmosis.
2. Disaccharides. Ashwagandha also utilizes disaccharides well. The most prevalent disaccharides are maltose, cellobiose, sucrose and lactose. Maltose is a product of starch hydrolysis, composed of glucose molecules connected by α-glycosidic bonds; fibrous disaccharides are also composed of glucose molecules, as is maltose, which are connected by β-glycosidic bonds, and fibrous disaccharides are the decomposition products of cellulose; sucrose contains one molecule of glucose and one molecule of fructose; and lactose, a constituent of milk, is composed of one molecule of glucose and one molecule of galactose. These disaccharides can be digested by maltase, cellobiase, sucrase, and lactase, respectively.
3. Polysaccharides. Starch is a plant polysaccharide, and the presence of amylase can be detected in the culture medium when starch is utilized as a carbon source for cultivation of ashwagandha. If the starch in the culture medium was replaced by other carbon sources, then the production of amylase decreased significantly or even ceased to be secreted. This proves that the amylase of Ashwagandha is an inducible enzyme and that it can only be synthesized when starch is present.
Cellulose, also a polysaccharide widely present in plant tissues, is the main carbon source available to wood-rotting fungi such as Ashwagandha. Ashwagandha produces a series of enzymes that break down cellulose, commonly known as cellulases, which readily degrade cellulose into glucose units. The first is "C1-cellulase", which mainly acts on insoluble crystalline cellulose to change it into soluble cellulose form, followed by "Cx-cellulase", which hydrolyzes soluble cellulose into monosaccharides. The third enzyme is cellobiose enzyme, hydrolyzes cellobiose to glucose, ashwagandha is able to synthesize this complex cellulase, making it capable of degrading cellulose of various structures, and it plays an important role in the cycle of matter in nature.
Lignin is a major constituent of perennial woody plants, and in nature it is second only to cellulose as an abundant organic polymorph, and it is a carbon source of wide potential for the saprophytic ashwagandha. Lignin is a closely related, structurally complex group of compounds with large relative molecular masses, which are polymers that include three substituted alcohols, which are coumarinol, pinacosideol, and mustelinol, and the inter-proportions of these subunits vary from species to species.
The structure of lignin prevents it from being utilized by most microorganisms, and it is mainly the wood-rotting mushrooms (white-rotting or brown-rotting) that are able to utilize lignin. Less is known about the mechanism of lignin degradation, except that the extracellular enzymes of wood-rotting mushrooms oxidize aromatic rings and aliphatic side chains to produce products of small relative molecular mass that are then utilized. In this process, phenolic oxidases such as laccase and peroxidase are indispensable enzymes in lignin degradation, and experiments have demonstrated that grass-rotting mushrooms (e.g., Straw Mushrooms) that cannot produce phenolic oxidases cannot degrade lignin.
(ii) Nitrogen. Nitrogen source is indispensable for the growth and development of ashwagandha, and its role is mainly to synthesize a variety of key cellular components, including ammonium, urea, amino acids, proteins, purines, nucleic acids, aminoglucans, chitin, and a variety of vitamins. Here we focus on the main types of nitrogen sources that ashwagandha can utilize and the factors that influence their utilization.
1. Urea. Urea, also known as urea, is a protein metabolite and is the main nitrogen-containing substance in mammalian urine. Molecular formula CO (NH2) 2, white crystals, containing 46% nitrogen, when heated above the melting point is decomposed into ammonia (NH3). Easily soluble in water, the solution is neutral reaction. In the production of ashwagandha, it is often used as the supplementary nitrogen source of solid culture medium (material), and its dosage is generally 0.1% to 0.2%. The addition of a large amount, toxic to the mycelium.
The test of urea uptake was carried out with 14C-urea in Ashwagandha, which proved that the urea transfer system exists. The first is an active transport system, which is an effect at low concentrations, and this system is induced by urea but inhibited by aspartate and glutamine. Alternatively, urea is able to be absorbed in a diffusive manner, which is an effect at high concentrations (500 micromol/liter), and this system is neither induced nor inhibited.
2. Ammonium fertilizers. There is a wide range of ammonium compounds, including ammonia bicarbonate (NH4HCO3, N=17.5%), ammonium sulfate [(NH4 )2SO4, N=21.2%], ammonia (NH3H2O, N=23%), and ammonium nitrate (NH4NO3, N=35%). When ammonium sulfate and ammonium nitrate are used as the nitrogen source, the final pH of the medium becomes very low, which is mainly due to the NO3- and SO42- remaining after NH4+ has been utilized, and is therefore called physiologically acidic salt. The pH can be adjusted to stabilize the reaction if appropriate amounts of buffer salts are added, such as gypsum and calcium carbonate.
When inorganic nitrogen sources such as ammonia bicarbonate and ammonium sulfate are used, the mycelium is able to utilize them but grows slowly. Ashwagandha does not utilize nitrate, which may be due to the inability of Ashwagandha to synthesize nitrate reductase.
3. Organic nitrogen. The gray tree flower is able to break down proteins in nature into amino acids that can be absorbed. In addition ashwagandha may absorb short-chained peptides, such as dipeptides and tripeptides, through a transfer system. Purines and pyrimidines have been shown to be utilized as a source of nitrogen by ashwagandha. Commonly used nitrogen sources are yeast paste, beef paste, peptone, casein, etc. These organic nitrogen sources can be degraded and absorbed and utilized by the mycelium faster, thus fast growth and higher biomass of Aspergillus mycelium when these types of composite nitrogen sources are used in the medium of the parent species.
(1) Peptone. Peptones are products of acid, alkali or enzymatic hydrolysis of natural proteins called peptones. Those made from animal meat are called meat peptones and pancreas peptones; those made from soybeans are called plant peptones or bean peptones. Biological reagent grade peptones, soluble in water, insoluble in alcohol, ether, do not solidify when heated, do not precipitate in saturated ammonium sulfate solution, no corrosive odor, no more than 2.5% residue after scorching, dry loss of no more than 8%, fat amount is not more than 0.5%, traces of reducing sugar amount. It is a good source of organic nitrogen for ashwagandha parent species culture medium, and the dosage is generally 0.2% to 1.5%.
(2) yeast paste. Is a yeast water-soluble components of the concentrate, viscous, dark brown, yeast flavor. Contains a variety of vitamins, amino acids and ash elements. Stimulating effect on the growth of gray tree flower, the preparation of the mother species culture medium commonly used, the use of concentration of 0.05% to 0.2%.
The production of commonly used bran, cornmeal, soybean meal, soybean cake powder, cotton kernel cake, animal feces and other nitrogen sources, these agricultural and sideline products in the processing of scraps in addition to rich protein, but also contains a variety of growth stimulating factors, not only more than peptone, yeast, beef paste and other valuable nitrogen source material effect is good, and the cost is also much lower.
(C) the carbon and nitrogen ratio of the culture material. The carbon and nitrogen ratio (C/N) of the culture material is appropriate, is an important indicator of its quality, directly affecting the occurrence of gray tree flower time and yield. The suitable C/N ratio of the culture material of gray tree flower is 15～20:1 at the stage of germination, and the nitrogen content is 1.6%～2%; the C/N ratio after mushroom emergence is 30～35:1. In other words, if the culture material contains 100 kg of carbon, it needs to be accompanied by 3～3.5 kg of nitrogen. Otherwise the bioassimilation cannot be carried out smoothly. The ratio of ashwagandha culture material and the amount of nitrogen added should strictly follow this requirement. If the nitrogen fertilizer is insufficient, it will obviously affect the yield of ashwagandha; if the nitrogen fertilizer is too much, it will not only cause waste, but also lead to difficulties in mushroom production due to the imbalance of carbon and nitrogen ratio. Within the appropriate range, the nitrogen content of the culture material is positively correlated with the yield.
Generally speaking, the natural materials used to cultivate ashwagandha have a high carbon to nitrogen ratio, such as wood chips at about 50:1; the carbon to nitrogen ratio of gramineous plants is 30-80:1. Obviously, the nitrogen in wood chips or straw is obviously insufficient, and thus nitrogen needs to be added when cultivating ashwagandha in order to make the culture material have a proper carbon to nitrogen ratio or nutritional balance.
Wood chips, crop residues and processing scraps of agricultural products are abundant, as long as the appropriate adjustment of its carbon and nitrogen content can be used for ashwagandha culture material. The relevant data of common raw materials are shown in Table 2-1.
Table 2-1 Carbon and Nitrogen Content and Carbon and Nitrogen Ratio of Commonly Used Raw Materials (C/N)
What is the effect of rice straw or bagasse on the cultivation of ashwagandha? Chen Guozhu et al. (1995) conducted a trial planting (Table 2-2), which proved that although Ashwagandha is a wood-rotting fungus, it can also be cultivated with rice straw and bagasse, and grows well, and the bioefficiency of the first tide of bag cultivation can reach about 40%, indicating that Ashwagandha has a wide range of adaptability to the culture material, and can be used in many kinds of crop by-products as culture material.
Table 2-2 straw, bagasse cultivation ashwagandha test results
Note: each formula plus soil 20, auxiliary materials 17 (bran 11%, cornstarch 5%, gypsum 1%).
(iv) Inorganic salts. In the process of ashwagandha cultivation, when the medium lacks certain inorganic elements, it will lead to slow growth of the bacterium or reduced reproduction ability, these inorganic elements are known as essential inorganic nutrients. The main functions of inorganic elements, in addition to constituting the constituents of cells, are to act as components of enzymes and maintain enzyme activity, and to regulate cellular osmotic pressure, hydrogen ion concentration, and redox potential. The amount of inorganic elements required by ashwagandha is very low compared to carbon and nitrogen sources, generally only a few hundred milligrams per liter of medium.
Based on the amount of inorganic nutrients required by the gray arborvitae, inorganic elements are divided into two categories: one is macronutrients, such as phosphorus (P), potassium (K), calcium (Ca), sulfur (S), magnesium (Mg), sodium (Na), etc.; and the other is the need for a small amount of micronutrients, including iron (Fe), zinc (Zn), copper (Cu), manganese (Mn), molybdenum (Mo), etc. The other is the need for a very small amount of microelements, including iron (Fe), zinc (Zn), copper (Cu), manganese (Mn), molybdenum (Mo). In addition to the need to add macronutrients, trace elements generally do not need to be added to the ashwagandha culture material. In the production formula, often add a small amount of minerals and inorganic salts are as follows:
1. Gypsum Gypsum's chemical name is calcium sulfate, the molecular formula for CaSO42 (H2O). The color is white, pink, yellowish or gray, transparent or translucent. It is plate-like, fibrous or fine-grained lumps with glassy luster. Heated to 150 ℃ dehydration mature gypsum, molecular formula for (CaSO4) 2H2O. mature gypsum in the production of ashwagandha, widely used as a solid medium (material) of the auxiliary material, the general dosage of 1% to 2%. Its role is to provide calcium, sulfur and other nutrients necessary for the growth of ashwagandha; can also accelerate the decomposition of organic matter in the raw material to promote the rapid release of soluble phosphorus, potassium; and can be neutralized and the acidity of the culture material.
Calcium ions enter the cell membrane of the gray tree flowers in the process of both active transportation and diffusion. It is currently believed that calcium ions are transported through the cell membrane in both inward and outward directions, and that this uptake and exocytosis is cyclic. The cyclic nature of calcium ion uptake is associated with cellular mitosis, with accumulation of calcium ions in the pre- and post-mitotic phases and calcium ion efflux in the middle phase of mitosis, whereas changes in uptake do not cause substantial changes in intracellular calcium ion concentration. Because calcium ions cause depolymerization of microtubule subunits in animal cells, it is thought that perhaps the cyclic influx of calcium ions during mitosis in Ashwagandha cells stimulates the release of calcium stored around the nucleus, and that calcium ions regulate mitosis through their effect on microtubule (spindle filament) formation.
2. Calcium carbonate. Calcium carbonate is made from ground limestone. In pure form, it is a white crystal or powder. The molecular formula is CaCO3. limestone powder is retained in the culture for a long time and effectiveness. Calcium carbonate is insoluble in water and the aqueous solution is slightly alkaline. When there is more carbon dioxide in the water, it can make it dissolve and generate soluble calcium bicarbonate. Ashwagandha mycelium in the appropriate water, nutrition and other environmental conditions, the discharge of carbon dioxide, calcium carbonate is absorbed by calcium bicarbonate to generate calcium bicarbonate, you can continue to ashwagandha to provide calcium nutrients, and acid and alkali buffer role. It is also a good material to supply calcium for ashwagandha pile and mulch. The amount used is generally 1% to 2%, if there is no calcium carbonate, lime can be used instead.
3. Lime The chemical name of quicklime is called calcium oxide (CaO), quicklime becomes slaked lime when it meets water, i.e., calcium hydroxide [Ca(OH)2]. It contains 2% to 20% gypsum, which can neutralize the excessive acid in the culture material and also supplement the calcium element in the culture material; it can also repel and kill some pests and stray bacteria. Lime is an alkaline substance, and in use generally do not mix with fertilizer to prevent reducing its effect. The amount used is 1% to 2%.
In addition, slaked lime also has a number of roles: ① in 5% of the carbolic acid solution with 1% of lime for environmental sterilization, the effect can reach 98.9%. ② raw material cultivation of edible fungi mixing, add 800 times the polymyxin dilution, and then add composite fertilizer and lime powder 1% each, can more effectively inhibit mold contamination. ③ When sour smell appears in the material, neutralize the treatment with lime water, the effect is very good. ④ segmented wood cultivation ashwagandha, with 15% ~ 20% lime water coated with segmented wood cross-section and skin, wood mold and other stray fungi have obvious inhibitory effect.
4. Calcium superphosphate Calcium superphosphate is also known as superphosphate lime. Commonly used for water-soluble, off-white to dark gray, or pinkish powder. It has an acidic odor, and the aqueous solution is acidic. The main chemical composition is calcium dihydrogen phosphate [Ca (H2PO4) 2H2O] and anhydrous calcium sulfate (CaSO4), phosphorus content (P2O5) 14% to 20%. Phosphorus is a very active element in the metabolism of microbial cells, and is a constituent of nucleic acids and phospholipids and their high-energy compounds ATP. Calcium superphosphate can provide phosphorus for the growth of ashwagandha, but also better to eliminate the ammonium flavor in the culture material and neutralize the high pH. calcium superphosphate is acidic, only used in the original and cultivated species of solid culture material, the dosage is generally 1% to 2%.
5. magnesium sulfate magnesium sulfate medicine commonly known as laxative salt. Molecular formula for MgSO47H2O, colorless or white, easy to weather crystals or white powder, with a bitter-salty taste, soluble in water. As magnesium ions (Mg3+) have an activating reaction to enzymes in microbial cells, for example, the action of hexokinase will be enhanced by the presence of magnesium ions. Magnesium sulfate is the most commonly used additive in various cultures of ashwagandha, with a general dosage of 0.03% to 0.2%.
Under normal conditions, the intracellular Mg2+ concentration of mycelium is directly proportional to the Mg2+ concentration in the culture medium, however, when the external Mg2+ concentration is increased, the intracellular Mg2+ concentration shows a certain degree of stability, for example, when the external concentration is increased up to 1,400-fold, the intracellular Mg2+ concentration is only increased by 1.5 to 3.3-fold. Inside the cell, Mg2+ participate in a large number of metabolic processes. Because they have negatively charged phosphate groups, Mg2+ is the most common ion that forms complexes with nucleotides. Important enzymes that require Mg2+ in ashwagandha are deoxyribonucleic acid (DNA) polymerase, chitinase, cellulase, and glutamine synthetase.
Mg2+ can affect the structure and function of cell membranes. Mg2+ is required for the synthesis of glycoproteins in the cell membrane, and Mg2+ can keep the membrane stable by keeping the protein components together, and Mg2+ can also inhibit transport by increasing the density of the membrane and making it less permeable.Mg2+ can affect a large number of biochemical processes, so it is not difficult to understand that Mg2+ affects growth and development. This is also the reason why magnesium sulfate is often added to the medium used to cultivate ashwagandha.
Sulfur is a component of proteins and vitamins, and is therefore required in large quantities by organisms. Amino acids such as methionine and cystine contain sulfur. The sulfhydryl (-SH) group in cystine plays an important role in stabilizing protein structure. Two cysteines are connected by disulfide bonds to form cystine. Disulfide bonds in proteins help maintain the folded conformation and affect the way proteins act in the cell. Sulfur is also involved in the construction of coenzymes, thiamine, biotin, coenzyme A, and zinc sulfate. Sulfate is the most prevalent form of sulfur utilized by ashwagandha.
The process of sulfur entry into proteins is divided into two main stages, in the first stage in which sulfate is initially activated by reacting with adenosine triphosphate (ATP) to form adenylyl sulfate, which then releases adenine, and a portion of the sulfur is reduced to sulfite, with the final free sulfite reduced to a divalent sulfur ion; in the second stage, the divalent sulfur ion is converted to organic sulfur, forming cysteine and methionine (methionine ). Although sulfate uptake is not governed by sulfite reduction and organic sulfur formation, the pathway from extracellular sulfur to intracellular cysteine and methionine is controlled by both precursors and end products. Ashwagandha can store excess sulfur in a variety of forms. For example, methionine, cysteine, and homocysteine in the sulfur assimilation pathway can all function as storage.
6. Potassium dihydrogen phosphate and dipotassium hydrogen phosphate Potassium dihydrogen phosphate molecular formula for KH2PO4, white fine-grained crystals, slightly acidic; dipotassium hydrogen phosphate molecular formula for K2HPO4, also white fine-grained crystals, slightly alkaline. Both of them can provide phosphorus and potassium nutrients for the ashwagandha, and can buffer the acidity and alkalinity in the culture medium (material). The general dosage of 0.1% to 0.2%.
Phosphorus in the form of PO43- is a component of many important macromolecules such as deoxyribonucleic acid (DNA), ribonucleic acid (RNA), phospholipids, as well as thiamine pyrophosphate, vitamin B12, coenzyme A (CoA). In addition, PO43- is involved in energy storage and conversion within the cell.
Phosphorus absorbed by ashwagandha is in the form of PO43-, which is secreted by ashwagandha as phosphatase, an enzyme that separates PO43- from phosphate compounds. It has been found in studies of gray arborvitae that the uptake of PO43- is active.
The main role of potassium ions in ashwagandha is to regulate osmotic pressure. The osmotic pressure of the cell is related to whether water in the medium enters the cell and provides the expansion pressure needed for growth, and uptake of potassium ions diminishes the difference between the osmotic pressures of the medium and the cytoplasm, altering the inhibitory effect of sodium ions on ashwagandha. Potassium ions also bind to proteins in the cell and activate enzymes, such as aldolase and pyruvate.
(v) Growth factors. The growth of ashwagandha requires some small amounts of organic substances, which do not act as nutrients like the carbon, nitrogen, and inorganic sources described earlier, but act as coenzyme components or function as coenzymes. Generally speaking, vitamins in the range of 0.01 to 0.1 micrograms per gram can play a role in promoting growth and development. There are also some organic substances do not belong to the vitamin class, but the low concentration of ashwagandha growth activity, called "organic growth factors", such as inositol, fatty acids, and growth hormone.
1. Thiamine (vitamin B1) ashwagandha needs is thiamine, its role is to regulate sugar metabolism, the active form is thiamine pyrophosphate (TPP). Thiamin pyrophosphate is a coenzyme for a number of enzymes in carbon metabolism, such as pyruvate decarboxylase, ketotransferase, and glutarate dehydrogenase. In pyruvate decarboxylase and ketoacyltransferase, thiamin pyrophosphate is an essential cofactor for subunit bonding of these enzymes. As an important cofactor, thiamin pyrophosphate has significant biological effects. If deficient, the mycelium is stunted in growth and development. In severe deficiencies, their growth stops completely. In addition, riboflavin, niacin and ascorbic acid all have a promoting effect on mycelial growth. Vitamins in potatoes, malt, yeast and rice bran in the content of more, so the preparation of culture medium with these materials can be used without adding. But most of the vitamins do not tolerate high temperatures, in the temperature above 120 ℃ is very easy to destroy, therefore, in the culture medium sterilization must prevent the temperature is too high or too long.
2. Biotin. Biotin is a growth factor second only to thiamine. Its active form is the carboxylase enzyme of certain active sites connected to lysine, this growth factor-lysine complex is called biocytin (biocytin). It functions as a coenzyme that transfers CO2 or carboxyl groups. Enzymes that require biocytin include pyruvate carboxylase, acetyl coenzyme A carboxylase, and urea carboxylase. Biotin has the same function in them, attaching the CO2 or carboxyl group before transferring it to the substrate.
In addition to the vitamins mentioned above, there are many other organic compounds that also affect the growth and development of ashwagandha, substances called growth regulators or growth factors, which differ from the vitamins in that they do not function as coenzymes. These regulators include some fatty acids, plant hormones, and some volatile substances. Fatty acids are substances that stimulate the growth and development of ashwagandha at low concentrations. Many studies have been conducted on the role of plant hormones or their analogs in the growth and development of ashwagandha, such as gibberellic acid (GA), indoleacetic acid (IAA), and naiacetic acid (NAA) can stimulate the germination of ashwagandha spores and the growth of mycelium. Indole acetic acid also stimulates an increase in mushroom body weight, cap diameter and stalk length. The most used hormone class is the plant growth hormone tricosanol, which can promote the growth of ashwagandha mycelium by treating it with low concentrations of tricosanol. The appropriate concentration of 0.5 ~ 1.0 g / ton of material, the effect of yield increase is obvious, the concentration is too high mycelial growth but inhibited.
Hebei University Biotechnology Development Company developed a "drop can be rich" yield enhancer, is a new type of compound amino acid growth promoter, the main ingredients for a variety of amino acids, nucleotides, trace elements and efficient active substances. Apply "Dicofon" to the cultivation of ashwagandha, add 0.2% of the amount of cultivation material can increase the biological conversion rate of 20%, and ashwagandha mycelium grows fast, early mushroom 5-7 days, the mushroom body is large, neat, low rate of disease mushrooms.