High-brittleness yellow flammulina velutipes HJ277 and application thereof

Abstract

The invention belongs to the technical field of edible mushroom cultivation, and particularly relates to a high-brittleness yellow flammulina velutipes HJ277 strain and application thereof. The technical problem to be solved by the invention is to cultivate a flammulina velutipes variety with better quality. According to the technical scheme, the yellow flammulina velutipes HJ277 with the high brittleness is obtained, and the preservation number of the yellow flammulina velutipes HJ277 is CCTCC (China Center for Type Culture Collection) No: M 20242219. The invention also provides a food which comprises the yellow flammulina velutipes, spores of the yellow flammulina velutipes or / and an extract of the yellow flammulina velutipes.

Description

Technical Field

[0001] The invention belongs to the technical field of edible fungus cultivation, and particularly relates to a high-brittleness yellow enoki mushroom HJ277 and an application thereof. Background Art

[0002] Flammulina velutipes, a member of the genus Flammulina in the family Tricholomataceae, is an aerobic, photophobic fungus. Its cap is spherical, with a thin, yellowish-brown margin and a slimy surface. It grows in clusters at the base. The dried product resembles daylily, hence the name. The surface of the velutipes is yellow, dark yellowish-brown, or cinnamon, with a slightly darker center and creamy yellow edges. The velutipes are white to creamy white or slightly pink, and vary in length. Enoki mushrooms are found in China, Japan, Russia, Europe, North America, and Australia. They often grow in clusters on rotten wood piles or roots in broad-leaved forests such as elm and willow trees, and occasionally on standing trees of various broad-leaved trees. Enoki mushrooms are a low-temperature edible fungus, with an optimum temperature of around 8°C. They are sensitive to moisture and require a high level of water. To prevent pests and diseases, humidity should be low when temperatures are high.

[0003] Enoki mushrooms are classified by color into three main categories: yellow, light-colored, and white. Because their fruiting bodies develop at relatively low temperatures, they are also known as winter mushrooms. Other common names include "gourmet mushroom," "puffer mushroom," "hairy-stemmed money mushroom," and "jelly mushroom." Enoki mushrooms belong to the Basidiomycota, Hymenomycetes, Agaricales, Tricholomataceae, and the genus Flammulina or Collybia. Wild Enoki mushrooms are all yellow, and farmers in the early years cultivated the yellow variety. Compared to white Enoki mushrooms, the traditional yellow variety looks dull, especially with its dark, almost black stem. In the late 1980s, the white variety bred in Japan entered the Chinese market and quickly replaced the yellow variety due to its attractive appearance and high yield. However, yellow Enoki mushrooms, due to their crisp texture and resistance to sticking to the teeth, have long been a popular snack ingredient. In the Sichuan and Chongqing regions, the yellow variety is also a popular hot pot ingredient. With increasingly diverse consumer demands, the yellow Enoki mushroom continues to have development potential.

[0004] Through hybrid breeding, my country has successively bred yellow varieties of Flammulina velutipes, such as "Sichuanjin No. 10" and "Sichuanjin No. 13". Their caps are dry, the stipes and caps are evenly colored, and the products look good. However, they still have the problem of getting stuck in teeth and being uncomfortable to chew. Further breeding of high-quality yellow Flammulina velutipes is needed. Summary of the Invention

[0005] The technical problem to be solved by the present invention is to cultivate varieties of golden needle mushrooms with better quality.

[0006] The technical solution of the invention is a high-brittle yellow enoki mushroom Flammulina velutipes HJ277, whose preservation number is CCTCC No: M20242219.

[0007] The fruiting body of HJ277 has a yellow, hemispherical cap with a diameter of 0.42 cm to 0.84 cm; the gills are white, slightly dense, and of unequal length; the stipe is yellow-white, 11.4 cm to 24.1 cm long, 0.17 cm to 0.44 cm in diameter, cylindrical, and has no hairs at the base.

[0008] The present invention also provides a food comprising the yellow enoki mushroom, spores of the yellow enoki mushroom and / or an extract of the yellow enoki mushroom.

[0009] Furthermore, the above-mentioned food is at least one of: fresh food, air-dried food, freeze-dried food, canned food or salted food.

[0010] Furthermore, the food is a health food or a functional food.

[0011] The present invention also provides a cultivation method of the yellow enoki mushroom HJ277, comprising the following steps:

[0012] S1. Mix the raw materials in a weight ratio of 53% cottonseed hulls, 9% broad bean hulls, 24% cotton residue, 1% rice straw, 7% corn flour, 2% bran, 1% phosphate fertilizer, 0.1% urea, 0.1% magnesium sulfate, 2% lime, and 0.8% gypsum, and adjust the pH to 6.0; sterilize; bag, cool, and inoculate;

[0013] S2. Place the inoculated bacteria bag in a culture room and culture for about 20 to 25 days. The relative humidity in the culture room should be controlled at 60 to 70% and the temperature should be controlled at 18 to 20°C.

[0014] S3. When the mycelium has grown to full size and has reached physiological maturity, scrape off the seed mass and old mycelium on the surface and then move it into the fruiting room;

[0015] S4. Adjust the temperature in the fruiting room to 12-15°C and the humidity to 95-98%. After the white mycelium is formed, keep the room temperature at 12-15°C, reduce the humidity to 90-95%, and set the CO2 concentration below 0.1%. Provide 200Lx light for 2 hours every day to promote the formation of primordia.

[0016] S5. When the primordium cap grows to the size of a mung bean, cover the mouth of a special bag for Enoki mushrooms with a vent hole and tie the mouth.

[0017] S6. When the enoki mushrooms grow to 12-15cm, they can be harvested.

[0018] In step S1, plastic bags with a width of 22 to 24 cm and a length of 45 cm are used for bagging, with each bag containing 1.2 to 1.25 kg of dry material.

[0019] Furthermore, in step S1, the mixture is sterilized by high pressure at 121° C. for 2 hours.

[0020] The present invention provides a strain of yellow enoki mushrooms with excellent appearance and crispness. While maintaining the advantages of high yield, high nutritional value, and ease of cultivation, this strain also boasts a crisp, tender texture and resistance to tooth decay, making it more readily accepted by consumers and promising commercial prospects. This application also provides a production method suitable for this strain, ensuring a stable production cycle, mushroom quality, and yield.

[0021] The yellow enoki mushroom Flammulina velutipes HJ277 of the present invention was deposited in the China Center for Type Culture Collection (address: Wuhan University, Wuhan, China) on October 15, 2024, with the deposit number: CCTCC No: M20242219. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 , the appearance picture of Rongjin 567.

[0023] Figure 2 , the appearance of Sichuan-Kinmen-7.

[0024] Figure 3 , monokaryons are paired and hybridized.

[0025] Figure 4 , the hyphae are locked together, as shown by the red circles in the figure.

[0026] Figure 5 , ISSR cluster diagram.

[0027] Figure 6 , HJ277 cap characteristics.

[0028] Figure 7 , comparison of HJ277 (right) and Rongjin 567 (left). DETAILED DESCRIPTION

[0029] The present invention is to select and breed high brittle yellow golden needle mushroom variety Rongjin 567 ( Figure 1 ) and the white enoki mushroom variety Sichuan Gold No. 7 ( Figure 2 ) conducted single-spore hybridization selection, using chewing texture, commercial properties, and yield as key indicators to identify a new strain of yellow enoki mushroom suitable for industrial production. The selected strain was also subjected to DUS assay and ISSR analysis, confirming it as a new variety.

[0030] At the same time, this applicant has also explored a cultivation method suitable for this strain, ensuring stable production cycles, mushroom quality, and yield. Specifically, the cultivation formula is as follows: 53% cottonseed hulls, 9% broad bean hulls, 24% cottonseed residue, 1% rice straw, 7% corn flour, 2% bran, 1% phosphate fertilizer, 0.1% urea, 0.1% magnesium sulfate, 2% lime, and 0.8% gypsum, with an initial pH of 6.0. The spawn is bagged in plastic bags 22-24 cm wide and 45 cm long, with 1.2-1.25 kg of dry material per bag. The bags are sterilized by autoclaving at 121°C for 2 hours. After cooling, the bags are inoculated. The inoculated bags are placed in a culture room for approximately 20-25 days, with a relative humidity of 60-70% and a temperature of 18-20°C. When the mycelium fills the bag and reaches physiological maturity, remove any surface clumps and old mycelium, then transfer the spawn to a fruiting room. The temperature in the fruiting room is adjusted to 12-15°C, and the humidity to 95-98%. After white mycelium forms, maintain the room temperature at 12-15°C and humidity at 90-95%. Increase ventilation appropriately to promote primordium formation. When the primordium cap grows to the size of a mung bean, cover it with a special enoki mushroom bag, leaving a vent hole and tie it. When the enoki mushrooms reach 12-15 cm in size, they can be harvested.

[0031] Example HJ277 breeding

[0032] 1. Isolation of spores and identification of monokaryotic hyphae

[0033] Cut off the stem of mature fresh fruiting bodies of Flammulina velutipes in a sterile environment, place the gill side flat on a sterilized culture dish, place it in a sterile room at 18-20℃, maintain the humidity, and collect spore prints overnight. Dilute the spore solution to a concentration of about 10 3 Each parent strain was cultured at a concentration of 100 μL. 100 μL of spore solution was evenly spread onto a PDA plate, inverted, and incubated in a 22-24°C incubator for 3-5 days. Observe spore germination. Once visible microcolonies appeared on the culture medium, use an inoculating needle to pick a single colony and transfer it to a PDA plate. A sterilized coverslip was inserted obliquely into the culture medium at the edge of the colony. Culture was continued in a 22-24°C incubator for 2-3 days. Once hyphae climbed onto the coverslip, the coverslip was removed and stained with lactophenol cotton blue. Monokaryons were observed under a 100x optical microscope. A total of 42 monokaryons of Rongjin 567 and 49 monokaryons of Chuanjin 7 were obtained.

[0034] 2. Monokaryotic hyphae pairing hybridization

[0035] Microscopically examined monokaryons were paired. Aseptically, using a sterile pipette, bacterial clumps approximately 0.8 cm in diameter were cut and cultured on PDA plates, 1 cm apart. The clumps were incubated at 22-24°C for 3-5 days. When a prominent fusion line formed between two fused colonies, hyphae were aseptically removed and transferred to another PDA medium. A sterile coverslip was inserted obliquely into the medium at the edge of the colony. The culture was incubated at 22-24°C for 2-3 days. Once hyphae climbed onto the coverslip, the coverslip was removed and the fusion was verified using the monokaryotic hyphae test. Any hyphae that exhibited a lock-like union were transferred to a PDA slant. Lock-like unions were observed in 300 of 2058 hybridization combinations, for a pairing success rate of 14.58%.

[0036] 3. Preliminary screening of bag material test

[0037] The cultivation formula is 53% cottonseed hulls, 9% broad bean hulls, 24% cottonseed residue, 1% rice straw, 7% corn flour, 2% bran, 1% phosphate fertilizer, 0.1% urea, 0.1% magnesium sulfate, 2% lime, and 0.8% gypsum. The initial pH is 6.0. Bag the spawn in plastic bags 22-24 cm wide and 45 cm long, with 1.2-1.25 kg of dry material per bag. Autoclave at 121°C for 2 hours. Cool the bags and inoculate. Place the inoculated bags in a culture room for approximately 20-25 days. Maintain a relative humidity of 60-70% and a temperature of 18-20°C. When the mycelium fills the bag and reaches physiological maturity, remove any surface clumps and old mycelium before transferring them to a fruiting room.

[0038] Adjust the temperature in the fruiting room to 12-15°C and the humidity to 95-98%. After white mycelium forms, maintain the room temperature at 12-15°C, reduce the humidity to 90-95%, and keep the CO2 concentration below 0.1%. Provide 200L of light for 2 hours daily to promote primordium formation. When the primordium cap grows to the size of a mung bean, cover it with a special bag for enoki mushrooms, leaving a vent hole and tie it. Harvest when the enoki mushrooms reach 12-15 cm.

[0039] Among the 300 hybrid offspring obtained, 261 strains produced mushrooms normally in the bag cultivation experiment. Based on the data of fruiting body maturity time, yield, cap diameter, stipe diameter and length, 9 better yellow enoki mushroom strains were preliminarily screened out, namely HJ86, HJ261, HJ108, HJ190, HJ206, HJ223, HJ241, HJ274 and HJ277.

[0040] The above nine yellow enoki mushroom strains were evaluated by two rounds of continuous cultivation and DUS testing (Guidelines for Plant Variety Specificity (Distinguishability), Uniformity and Stability Testing of Flammulina velutipes) (Table 1).

[0041] The DUS test results showed that the HJ277 strain had the highest average yield of 303.1g, followed by HJ206 with 302g; considering the comprehensive indicators such as the cap, stipe color, shape, and the degree of involution of the stipe edge, the HJ108, HJ206, HJ241 and HJ277 strains performed better.

[0042] Table 1 DUS test data of yellow enoki mushroom strain

[0043]

[0044] ISSR molecular markers and cluster analysis were performed on the nine Flammulina velutipes strains evaluated by the DUS test. The results showed that the nine strains could be divided into five major groups when classified at 75% similarity. Cluster I included strains HJ108, HJ206, and HJ274, and cluster II included strains HJ170, HJ223, and HJ261.

[0045] The brittleness of the fruiting bodies of the two rounds of cultivation was measured (Table 2). The average brittleness of the parent control Rongjin 567 was 347g, and the brittleness of Chuanjin 7 was 211g. Among the other strains, the brittleness of the yellow Flammulina velutipes strain HJ277 was the highest, at 345g. Figure 4 ) has good commercial application prospects and was sent to the China Center for Type Culture Collection (CCTCC) for preservation.

[0046] Table 2 Cultivated strain brittleness test values

[0047] serial number Crispness (g) HJ86 149 HJ261 223 HJ108 231 HJ190 171 HJ206 61 HJ223 206 HJ241 217 HJ274 208 HJ277 345

[0048] The HJ277 strain was tested for specificity, consistency, and stability over two consecutive growth cycles. Population traits were observed from the entire plot and mixed samples at 70% maturity. 60 fruiting bodies of each strain were sampled for individual site observation.

[0049] HJ277 mycelium cultivation takes 20 to 25 days, primordium formation takes 10 to 14 days, and fruiting bodies develop in 14 to 18 days, with a moderate to large number of fruiting bodies. The cap is yellow with a strongly involuted edge. The diameter ranges from 0.42 to 0.84 cm, the height ranges from 0.32 to 0.56 cm, and the thickness ranges from 0.21 to 0.50 cm. The stipe of HJ277 has a uniform color and low adhesion. The stipe is 11.4 to 24.1 cm long and 0.17 to 0.44 cm in diameter. Therefore, HJ120 has good commercial application prospects and has been deposited with the China Center for Type Culture Collection (CCTCC).

Claims

1. A highly brittle yellow enoki mushroom, Flammulina velutipes HJ277, whose accession number is CCTCC No: M20242219.

2. A food, characterized in that: The invention comprises the yellow enoki mushroom, the spores of yellow enoki mushroom and / or the extract of yellow enoki mushroom as claimed in claim 1.

3. The food according to claim 2, characterized in that: The food is at least one of fresh food, air-dried food, freeze-dried food, canned food or salted food.

4. The food according to claim 2, characterized in that: The food is a health food or a functional food.

5. The method for cultivating yellow enoki mushroom according to claim 1, characterized in that: The steps include: S1. Mix the raw materials in a weight ratio of 53% cottonseed hulls, 9% broad bean hulls, 24% cotton residue, 1% rice straw, 7% corn flour, 2% bran, 1% phosphate fertilizer, 0.1% urea, 0.1% magnesium sulfate, 2% lime, and 0.8% gypsum, and adjust the pH to 6.0; sterilize; bag, cool, and inoculate; S2. Place the inoculated bacteria bag in a culture room and culture for about 20 to 25 days. The relative humidity in the culture room should be controlled at 60 to 70% and the temperature should be controlled at 18 to 20°C. S3. When the mycelium has grown to full size and has reached physiological maturity, scrape off the seed mass and old mycelium on the surface and then move it into the fruiting room; S4. Adjust the temperature in the fruiting room to 12-15°C and the humidity to 95-98%. After the white mycelium is formed, keep the room temperature at 12-15°C, reduce the humidity to 90-95%, and set the CO2 concentration below 0.1%. Provide 200Lx light for 2 hours every day to promote the formation of primordia. S5. When the primordium cap grows to the size of a mung bean, cover the mouth of a special bag for Enoki mushrooms with a vent hole and tie the mouth. S6. When the enoki mushrooms grow to 12-15cm, they can be harvested.

6. The cultivation method according to claim 5, characterized in that: In step S1, plastic bags with a width of 22 to 24 cm and a length of 45 cm are used for bagging, with each bag containing 1.2 to 1.25 kg of dry material.

7. The cultivation method according to claim 5, characterized in that: In step S1, the mixture is sterilized by high pressure at 121° C. for 2 hours.

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