Auricularia auricula strain Tianyun No.1 and application thereof

Abstract

The invention belongs to the field of microorganisms, and relates to a black fungus strain Tianyun No.1 and application thereof. The invention provides a new black fungus strain which is brand new and successfully preserved, and the new black fungus strain is rich in sporocarp nutrition, is rich in various bioactive substances, and can be better applied to the aspects of food and medicines; a fresh product of the novel black fungus strain is ear-shaped, light yellow brown, semitransparent colloid, good in elasticity, smooth, free of veins on the ventral surface (pregnant surface) and the back surface (sterile surface), good in slicing property, basically free of ear roots, 6.7 * 4.5 cm in average size and 1.43 mm in average thickness; after being dried, the product is strongly shrunk into cutin which is hard and crisp, the back surface is off-white and has short villus, and the ventral surface is black brown and smooth; after the dried ears are fully soaked and loosened, the morphological characteristics of the dried ears are almost the same as those of fresh ears, and the soaking and loosening rate is not lower than 13.0; during stir-frying, a sound like setting off a firecracker can be made; the high-temperature-resistant and impurity-resistant cultivation medium is high in yield, tender and crisp in taste, fresh, fragrant, glutinous and tasty, is deeply loved by people, is suitable for cultivation of various agricultural and forestry byproducts such as weed tree sawdust, mulberry branch sawdust and corncobs (stalks), is high in high-temperature resistance and impurity resistance, and has good popularization and application and market prospects.

Description

Technical Field

[0001] The present application belongs to the field of microorganisms and relates to a black fungus strain "Tianyun No. 1" and its application. Background Art

[0002] Black fungus (Auricularia auricular) belongs to the genus Auricularia of the phylum Eumycota, subphylum Basidiomycotina, class Hymenomycetes, order Auriculariales, family Auriculariaceae, kingdom Mycota, in fungal taxonomy. It is also known as black fungus, fungus, cloud fungus, etc.

[0003] Black fungus (Auricularia heimuer) is a typical colloid fungus, containing protein, dietary fiber, polysaccharides, amino acids, melanin, flavonoids, polyphenols, and 48 macro- and trace elements such as iron, zinc, calcium, and manganese. It is a nutritious edible fungus that is both medicinal and edible. Modern medical research has found that black fungus has multiple functions such as antioxidant and antibacterial activity and prevention of cardiovascular diseases. Actual clinical trials have proven that black fungus has the effect of preventing and treating coronary heart disease and atherosclerosis. Depending on the variety, origin, cultivation medium, and management method, the various nutrients and functional active substances contained in black fungus are different.

[0004] Black fungus produced in Guangxi is commonly known as cloud fungus. The main production area of ​​wild cloud fungus is in Baise. It belongs to the Auriculariales, Auriculariaceae, and Auricularia genus. It is a large colloid fungus that can be used for both medicine and food. As of the beginning of the 21st century, the production of Baise cloud fungus was still in a wild or semi-wild state, and its output was far from meeting the market demand. Therefore, separating and domesticating wild Baise cloud fungus and realizing artificial cultivation is the inevitable direction for the development of Baise cloud fungus.

[0005] With the development of biology and biotechnology, researchers hope to find more excellent black fungus strains with promotion and application value. Summary of the invention

[0006] In view of the shortcomings of the prior art, the present application provides a fresh black fungus strain, which has a bioconversion rate of not less than 130% and excellent appearance, and is an excellent black fungus strain with promotion and application value. The specific contents are as follows:

[0007] A black fungus strain, which was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration Committee (No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing) on ​​March 22, 2024, with a deposit number of CGMCC No. 41270, and a classification name of black fungus Auricularia auricularia. The black fungus strain (Tianyun No. 1) is an excellent strain obtained by sampling from Xiangyang Town, Tiane County, through separation, purification, qualitative domestication.

[0008] Furthermore, the black fungus strain includes spores, mycelium and fruiting bodies of the black fungus strain.

[0009] The application of the black fungus strain in food production.

[0010] The application of the black fungus strain in medicine preparation.

[0011] Furthermore, the black fungus strain includes spores, mycelium and fruiting bodies of the black fungus strain.

[0012] The fresh fruiting bodies of the black fungus strain (Tianyun No. 1) described in the present application are rich in nutrition and various biologically active substances. The fresh product is ear-shaped, light yellow-brown, translucent, colloid, elastic, smooth, and has no veins on the ventral side (pregnant side) and the dorsal side (infertile side). It has good sliceability and basically no ear roots (see Fig.14 ), average size 6.7×4.5cm, average thickness 1.43mm; after drying, it shrinks strongly into keratin, hard and brittle, with grayish white and short hairs on the back and dark brown and smooth on the ventral side; after the dried ears are fully soaked, their morphological characteristics are almost the same as those of fresh ears, and the soaking rate is not less than 13.0; when stir-fried, it will make a sound like setting off firecrackers; the taste is tender, crisp, fresh, fragrant, glutinous and refreshing, and is deeply loved by people. It is suitable for cultivation with a variety of agricultural and forestry by-products such as sawdust, mulberry branch chips, corn cobs (stalks), etc. It has strong resistance to high temperature and impurities, and the bioconversion rate of fresh ears is not less than 130%. It has excellent appearance and is an excellent black fungus strain with promotion and application value. It can be well used in food and medicine.

[0013] The method for cultivating black fungus strains controls the temperature range to be 15-35° C. and the pH value range to be 5.5-6.5.

[0014] Furthermore, the temperature range is 25-28°C.

[0015] Furthermore, the pH value range is 6.0.

[0016] Furthermore, the water content of the solid culture medium used is 65%.

[0017] Furthermore, the culture medium used was as follows: 200 parts of potatoes (peeled), 20 parts of glucose, 3.0 parts of potassium dihydrogen phosphate, 1.5 parts of magnesium sulfate, 1000 parts of water, and natural pH.

[0018] Beneficial effects of this application:

[0019] (1) This application provides a new strain of black fungus that was completely isolated and successfully preserved.

[0020] (2) The new black fungus strain provided in the present application has a fresh fungus bioconversion rate of not less than 130% and excellent appearance, and is an excellent black fungus strain with promotion and application value.

[0021] (3) The new black fungus strain provided in the present application has a fruiting body rich in nutrients and various biologically active substances such as polysaccharides and pectin, and can be well applied in food and medicine.

[0022] (4) The new strain of black fungus given in the present application is ear-shaped, light yellow-brown, translucent, elastic, smooth, with no veins on the ventral (pregnant) and dorsal (infertile) sides, good sliceability, and basically no ear roots. The average size is 6.7×4.5 cm and the average thickness is 1.43 mm. After drying, it shrinks strongly into keratin, is hard and brittle, has a grayish white back with short hairs, and is dark brown and smooth on the ventral side. After the dried ear is fully soaked, its morphological characteristics are almost the same as those of the fresh ear, and the soaking rate is not less than 13.0. When stir-fried, it will make a sound like setting off firecrackers. It has a tender, crisp, fresh, fragrant, glutinous and refreshing taste, is deeply loved by people, and has excellent edible prospects. It is suitable for cultivation with a variety of agricultural and forestry by-products such as miscellaneous wood chips, mulberry branch chips, and corn cobs (stalks). It has strong resistance to high temperatures and weeds, and has good promotion and application and market prospects.

[0023] (5) The new black fungus strain provided in the present application is a wide temperature and high temperature resistant variety, with a temperature range of 13-35°C. When the temperature is close to 36°C, its mycelium is still white and strong as before, and the fruiting body does not suffer from diseases such as ear flow (see Fig.15 ), and the mushrooms can continue to grow in the future without being affected by high temperatures (see Fig.16 ), while the ear temperature range of existing varieties is generally 13-28℃.

[0024] (6) The new black fungus strain given in the present application was cultured using mulberry branch particles (diameter 2-3mm, length 2-3mm) and Cyclobalanopsis glauca particles (diameter 2-3mm, length 2-3mm) and then stored in a refrigerator at 4-6°C for more than 13 years. There was no significant change in its performance indicators. However, there has been no report on the long-term preservation of other varieties using this method. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1This is a photo of the fresh fruiting body of Tianyun No. 1.

[0026] Figure 2 This is a photo of the dried fruiting body of Tianyun No. 1.

[0027] Figure 3 This is a photo of fresh fruiting bodies of Black 29.

[0028] Figure 4 Photo of the culture characteristics of Tianyun No. 1 mycelium.

[0029] Figure 5 The electrophoresis patterns of various black fungus strains are as follows: 1-Baiyun No. 6, 2-Guiyun No. 3, 5-Baiyun No. 9, 6-Tianyun No. 1.

[0030] Figure 6 This is the developmental tree of each black fungus species, including: E1-Baiyun No. 6, E2-Guiyun No. 3, E5-Baiyun No. 9, E6-Tianyun No. 1.

[0031] Figure 7 This is the identification result of Baiyun No. 9 (E6), which was identified as a black fungus strain.

[0032] Figure 8 It is the antagonistic line between Tianyun 1 and Baiyun 9.

[0033] Fig. 9 It is the antagonistic line between Tianyun 1 and Guiyun 3.

[0034] Fig.10 It is the antagonistic line between Tianyun No. 1 and Black Fungus 916.

[0035] Fig.11 It is the antagonistic line between Tianyun 1 and Baiyun 6.

[0036] Fig.12 It is the antagonistic line between Tianyun No. 1 and Black 29.

[0037] Fig.13 It is the antagonism line between Tianyun 1 and Montenegro 15.

[0038] Fig.14 These are comparison photos of the appearance differences between the Tianyun No. 1 strain of this application and the comparative Hei No. 29 strain.

[0039] Fig.15 This is the growth condition of the Tianyun No. 1 strain of the present application when the temperature is close to 36°C.

[0040] Fig.16 This is the mushroom production situation of the Tianyun No. 1 strain of the present application when the temperature is close to 36°C. DETAILED DESCRIPTION

[0041] The present application is further described in detail below in conjunction with specific embodiments, which are intended to explain the present application rather than to limit it.

[0042] Example:

[0043] 1. Relevant information of the strain: including function, classification, properties, etc.

[0044] ①Classification status: Auricularia auricularia belongs to the genus Auricularia of the phylum Eumycota, subphylum Basidiomycotina, class Hymenomycetes, order Auriculariales, family Auriculariaceae, kingdom Mycota in fungal taxonomy. It is also known as black vegetable, fungus, cloud ear, etc.

[0045] ② Characteristics (including fruiting body morphology, mycelium culture characteristics, biological characteristics, cultivation characteristics, etc.):

[0046] a. Morphological characteristics of fruiting bodies The black fungus produced in Guangxi is commonly known as cloud ear. The main production area of ​​wild cloud ear is in Baise area. It belongs to the large colloid fungus for both medicine and food of the order Auriculariales, family Auriculariaceae, and genus Auricularia. The Tianyun No. 1 of the present application is an excellent strain obtained by sampling from Xiangyang Town, Tian'e County, through separation, purification and qualitative domestication. The fresh fruiting body is ear-shaped, light yellow-brown, translucent colloid, elastic, smooth, with no veins on the ventral side (pregnant side) and the dorsal side (infertile side), good sliceability, basically no ear roots, with an average size of 6.7×4.5cm and an average thickness of 1.43mm; after drying, it strongly shrinks into keratin, is hard and brittle, has a grayish white back with short villi, and a dark brown and smooth ventral side; after the dried ear is fully soaked and loosened, its morphological characteristics are almost the same as those of the fresh ear; when stir-fried, it will make a sound like setting off firecrackers; the taste is smooth and not greasy, crisp and refreshing, with a pleasant fragrance (below Figure 1 After drying, it shrinks strongly into keratin, which is hard and brittle. The back is off-white with short hairs, and the ventral side is dark brown and smooth (below Figure 2 ); It is different from the main variety of black fungus in the main producing area of ​​Northeast China, Black 29, which is black, with obvious grid patterns (vein patterns, commonly known as "tendons") on the back (sterile surface) and a hard feel (below Figure 3 ).

[0047] Tianyun No. 1 mycelium was cultured in a conventional comprehensive potato medium [200g potatoes (peeled), 20g glucose, 3.0g potassium dihydrogen phosphate, 1.5g magnesium sulfate, 10mg vitamin B1, 1000ml water, natural pH.] The culture characteristics are as follows: the matrix mycelium is grayish white, sparse and weak in the early stage, gradually dense and strong in the later stage, the colony is round, the aerial mycelium is sparse, and there is basically no wall climbing phenomenon (lower Figure 4). BGI identified the strain of the present application as a black fungus variety ( Figure 5-7 ); It has a significant antagonistic reaction with the black fungus strains Hei 29, 916, Heishan 15, Baiyun 6, Guiyun 3, and Baiyun 9, which are currently applying for a patent (below) Figure 8-13 )

Note: Individuals with significantly different genetic backgrounds will form a clear dividing line in the junction area, which is the antagonistic reaction. The antagonistic reaction is a specific manifestation of somatic cell incompatibility. It has been used in the preliminary study of fungal kinship due to its rapid, convenient and intuitive advantages. It has been used as the main means of fungal species identification.

[0048] The black fungus strain of the present invention is obtained by screening, domestication and cultivation of wild black fungus in Guangxi, and the screening and domestication methods are as follows:

[0049] (1) 23 wild black fungus fruiting bodies collected from Xiangyang Town, Tian'e County, Guangxi (106°58'\25°5'\948m) were used to obtain 17 pure strains through tissue separation and purification, and all 17 pure strains were identified as wild black fungus (cloud fungus) strains through direct ear extraction;

[0050] (2) 17 wild black fungus (cloud ear) strains were subjected to an ear evaluation test. Based on the ear performance, ear piece color, ear root size, ear piece size and thickness, anti-impurity ability, foaming rate, and bioconversion rate as indicators, the strain with good ear performance, basically no ear root, large and thick ear pieces, strong anti-impurity ability, and high foaming rate and bioconversion rate was selected as the mother strain Au-1;

[0051] (3) The mother strain Au-1 was cultivated in artificial bags and acclimated and screened in different seasons. Based on the indicators in (2) and temperature adaptability and temperature resistance, 15 strains with better indicators, wide temperature adaptability and good high temperature resistance were screened and recorded as F1 generation;

[0052] (4) The F1 generation strains were cultivated in artificial bags and acclimatized and screened in different seasons. Referring to all the indicators in (3), 7 better strains were screened out and recorded as the F2 generation;

[0053] (5) The F2 generation strains were cultivated in artificial bags and acclimatized and screened in different seasons. Referring to all the indicators in (3), three excellent strains were selected and recorded as the F3 generation;

[0054] (6) The F3 generation strains were cultivated in artificial bags and domesticated and screened in different seasons. Referring to all the indicators in (3), an excellent strain with good ear growth performance, basically no ear roots, no veins, light yellow-brown, translucent, good elasticity, an average ear piece size of 6.7×4.5 cm, an average thickness of 1.43 mm, a wide range of material adaptability, strong resistance to impurities, a wide temperature adaptability, high temperature resistance, a swelling rate of not less than 13% and a bioconversion rate of more than 130% was selected and named "Tianyun No. 1" after the place of origin.

[0055] Among them, in step (1) to step (6), the method for separating and purifying the tissue of the fruiting body is as follows: select healthy and disease-free black fungus fruiting bodies, wipe the surface of the black fungus fruiting bodies lightly with gauze soaked in 75% Vol alcohol solution, put them into a sterilized culture dish and place them in a clean bench, turn on the ultraviolet lamp and the fan at the same time, after 1 hour, when the moisture on the surface of the ear piece has dried and shrunk by 1\3~1\5, use tweezers and scissors burned by an alcohol lamp and cooled to cooperate to cut a 2mm×2mm tissue block near the ear root, insert it into a PAD slant culture medium without water accumulation, and culture it at a constant temperature of 25~28℃ until the mycelium grows to 1\2~1\3 of the slant, pick the front mycelium, transfer it to a fresh slant culture medium for culture, and continue to pick the front mycelium and transfer it to a fresh slant culture medium for culture in the same way. Repeat this method for multiple selection and transplantation and combine it with microscopic examination until a pure strain is obtained. The formula of the test tube slant culture medium is: 1000ml of 20% peeled potato juice, 20g of glucose, 3g of potassium dihydrogen phosphate, 1.5g of magnesium sulfate, 15-17g of agar powder, and natural pH.

[0056] Among them, in step (1) to step (6), the black fungus cultivation and domestication method adopts bag-type cooked material cultivation, and the cultivation steps are as follows:

[0057] (1) Preparation of mushroom bags: Use 17cm×33cm×0.05mm low-density polyethylene edible fungus bags, put the culture material into the bag according to the conventional method, make mushroom bags, and then sterilize: When using normal pressure steam sterilization, the center material temperature should reach above 98℃ within 4 to 5 hours after the cold air is exhausted, and continue to maintain for 12 to 15 hours; when using high-pressure steam sterilization, the pressure reaches 0.15MPa after the cold air is exhausted, and the material temperature is maintained at 126℃ for 3 to 3.5 hours. After pre-cooling and forced cooling to a material temperature of 25 to 30℃, the sterilized mushroom bags are sent to the clean inoculation room and inoculated according to the requirements of the aseptic operation procedures.

[0058] The planting material consists of the following components in parts by weight: 0-78 parts of miscellaneous wood chips, 0-88 parts of mulberry branch chips, 0-90 parts of cottonseed hulls, 0-73 parts of corn cobs, 0-73 parts of bagasse, 7-15 parts of wheat bran, 0-3 parts of corn flour, 1-2 parts of soybean flour, 0-1 part of edible-grade glucose, 0.5-1 part of gypsum, and 0.5-1 part of lime.

[0059] (2) Mycelium culture: The inoculated fungus bags are promptly sent to the culture room and placed on shelves. The indoor environmental parameters are: temperature 25-28°C, humidity 55-65%, dim light, and fresh air. The culture time is 40-45 days.

[0060] (3) Opening the ear holes: The cultured mushroom bags (sticks) are transported to the place where the opening is to be made. The opening site should be clean and sanitary, disinfected with 0.1% potassium permanganate solution or carbendazim, and covered with a layer of clean plastic film. The opening device (machine) and the transport frame and other places that come into contact with the mushroom bag should also be disinfected. If an electric opening machine is used, care should be taken not to splash the solution onto the power supply and the motor. Then, 1 to 2 large barrels should be prepared, each containing about half a barrel of potassium permanganate solution for surface disinfection of the mushroom bag. When everything is ready, the mushroom bag opening operation can be officially started. Generally, three people are required to cooperate. One of them puts the mushroom bag to be opened into the disinfectant for surface disinfection, one person is responsible for opening, and the third person is responsible for receiving the bag and placing it in the transport frame. The mushroom bag is opened vertically, and the cutter wheel can be made of round nails, triangular nails, or straight nails. The hole spacing is 0.8 cm to 5 cm, the hole diameter is 0.3 cm to 0.6 cm, and the hole depth is 0.4 cm to 0.8 cm.

[0061] (4) Ear bud induction: Arrange the opened mushroom bags in a triangle shape on the ear-growing rack or on the surface of the ear-growing greenhouse, with a spacing of 15cm to 20cm between the mushroom bags and a shelf height of 45cm to 50cm. Management methods: ① Natural climate ear-growing greenhouse: Put down the surrounding film and close the greenhouse door, and use the humidification equipment to automatically control the air humidity in the greenhouse to always be around 95%, the light intensity is not less than 500lx, and whether to put down the surrounding film and close the greenhouse door depends on the temperature and CO2 content in the greenhouse. Principle: If the greenhouse temperature exceeds 28℃, the surrounding film cannot be put down and the greenhouse door cannot be closed. Otherwise, the surrounding film can be put down and the greenhouse door can be closed; ② Indoor shelf ear-growing: Use relevant equipment to automatically control the indoor temperature to always maintain 25℃±2℃, the light intensity is not less than 500lx, the air humidity is around 95%, and the CO2 content is within 500ppm. After 5 to 7 days of management, the primordium has basically begun to differentiate, and some may have formed ear buds. After 10 to 12 days, ear buds are formed and more than 80% of them differentiate into ear pieces, then enter the ear piece growth, which is the critical water management stage.

[0062] (5) Management of ear growth: When most of the ear pieces grow to a size of 20mm, a sprinkler system should be used for water management. ① Natural climate ear growth greenhouse: When the temperature in the greenhouse is higher than 28℃, do not spray water. If the ear pieces are not dry, do not spray water. ② Indoor shelf ear growth: Use corresponding equipment to control the room to always maintain a temperature of 20-25℃ and clean air. When the dryness of the ear pieces reaches more than 80%, you can start spraying water.

[0063] The water temperature should be controlled between 20-25℃. The number of times and duration of water spraying depends on the state of the ear pieces. Each time, the ear pieces should be sprayed until they are fully expanded, like the state of dry wood ear fully soaked in water. Since dry wood ear pieces have a process of absorbing water, in order to save water, spray for 20-30 minutes, stop spraying for about 15 minutes, spray for another 20-30 minutes, and stop spraying for about 15 minutes. Repeat this 3-5 times until the ear pieces are fully expanded, and then wait for the next spray.

[0064] Follow the above principles to manage the ear pieces during their growth period until they are ready to be harvested. Under normal circumstances, this stage takes 10 to 15 days.

[0065] (6) Harvest at the right time

[0066] The fruiting bodies that have grown and matured should be harvested in time, and they should be harvested frequently and carefully to achieve the goal of high yield and high quality. The best harvesting period is when the ear pieces are 80% to 90% ripe. At this time, the ear pieces are of good quality and have weight. The standard for the maturity of wood ear is that the color turns light, the ear roots change from large to small, and the ear pieces stand upright. When the ear pieces are fully unfolded, the edges are wrinkled and thin, the ear roots shrink, and the back of the ear is light white, the spores are about to be ejected, and watering should be stopped before harvesting. The principle of picking ears is to harvest in time, pick the big ones and leave the small ones. If the ear pieces are not unfolded and have no commercial value, do not pick them, and keep them for the next harvest. Harvesting method: You can use a blade to cut them at the base of the ear without culture medium to keep the ear pieces clean, or you can hold the ear pieces by hand and twist them off at the base.

[0067] (7) Drying in time

[0068] After harvesting, the auricles should be dried in time. The methods include natural drying and hot air drying with drying equipment. The moisture content can be below 11%. Since Guangxi's special black fungus is rich in polysaccharides and pectin, it should be turned frequently during drying, especially at the beginning, to prevent the auricles from sticking together or sticking to the utensils. For the harvesting and drying method, a drying rack should be set up in advance. The rack should be 0.8 to 1.0 m above the ground, and a small pole should be placed on top. Screens and shade nets should be laid on top to facilitate ventilation. The natural light ventilation and cooling method not only improves the drying quality, but also solves the problem of fungus rotting on rainy days. Before drying, use scissors to cut off the culture medium at the root of the fungus. When drying, the leaves should be on top and the roots should be on the bottom. In addition, the clustered cloud ears must be torn into single pieces before drying. If it rains, cover it with rainproof material. The rainproof material should be covered above the drying rack, leaving a certain space for air circulation. Do not cover it directly on the ear pieces to prevent the ear pieces from being crushed and deformed, which will affect the quality of the fungus.

[0069] Finally, it should be pointed out that the above embodiments are only representative examples of the present application. Obviously, the technical solution of the present application is not limited to the above embodiments, and there may be many variations. All variations that can be directly derived or associated with the contents disclosed in the present application by a person of ordinary skill in the art should be considered as the protection scope of the present application.

Claims

1. A black fungus strain, characterized in that It is deposited in the General Microbiology Center of China Microorganism Culture Collection Administration with the deposit number CGMCC No.41270.

2. The black fungus strain according to claim 1, characterized in that The black fungus strain comprises spores, mycelium and fruiting bodies of the black fungus strain.

3. Use of the black fungus strain according to claim 1 in food production.

4. Use of the black fungus strain according to claim 1 in drug preparation.

5. The use according to claims 3 and 4, characterized in that: The black fungus strain comprises spores, mycelium and fruiting bodies of the black fungus strain.

6. The method for cultivating the black fungus strain according to claim 1, characterized in that: The controlled temperature range is 15-35°C and the pH value range is 5.5-6.

5.

7. The method for cultivating black fungus strain according to claim 6, characterized in that: The temperature range is 25-28°C.

8. The method for cultivating black fungus strain according to claim 6, characterized in that: The pH range is 6.

0.

9. The method for cultivating black fungus strain according to claim 6, characterized in that: The water content of the solid culture medium used was 65%.

10. The method for cultivating black fungus strain according to claim 6, characterized in that: The culture medium used was as follows: 200 parts of potatoes (peeled), 20 parts of glucose, 3.0 parts of potassium dihydrogen phosphate, 1.5 parts of magnesium sulfate, 1000 parts of water, pH natural.

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