New mushroom strain and application and cultivation method thereof

Abstract

The invention belongs to the field of microorganisms, and relates to a novel mushroom strain and application and a cultivation method thereof. The invention provides a novel lentinula edodes strain 'Yao xiang No.1' (Lentinula edodes), which has been preserved in China General Microbiological Culture Collection Center (address: Institute of Microbiology, Chinese Academy of Sciences, No.3, No.1 Yard, Beichen West Road, Chaoyang District, Beijing) on January 13, 2025, and has a preservation number of CGMCC NO.41776. Compared with existing strains which are popularized and applied in the public market and are subjected to modern breeding methods such as hybridization, physical mutagenesis and cell fusion, sporocarp pilei of the strain are thin and soft, stipes are thin and long, pleat intervals are sparse, shiitake mushrooms are fresh, sweet and pure in taste during cooking, rich and elegant in fragrance and refreshing in heart, and the shiitake mushrooms feel refreshing after being dried and smelling fragrance, so that the shiitake mushrooms have good taste. The Yao-xiang No.1 sporocarp product is more suitable for walking in the middle-and-high-end market.

Description

Technical Field

[0001] This application belongs to the field of microorganisms and relates to a new Lentinula edodes strain, its application and cultivation method. Background Art

[0002] Lentinula edodes is also known as fragrant mushroom, fragrant xun, flower mushroom, flower xun, fragrant letter, shiitake mushroom, winter mushroom, thick mushroom, etc.; the fruiting bodies of wild Lentinula edodes are mostly solitary, while those of artificially cultivated Lentinula edodes by bag cultivation can be solitary, clustered or growing in groups, and the fruiting bodies are medium to slightly large. The mature fruiting body is umbrella-shaped and consists of three parts: the pileus, the lamellae and the stipe. Pileus: Located at the top of the stipe, round, with a diameter generally of 3 - 15 cm, and up to 20 cm in some cases. It is hemispherical when young and then becomes flat to slightly flat. The surface color ranges from rhombic, light brown, dark brown to dark cinnamon color, and there are often dark scales in the middle, while there are often dirty white hairy or flocculent scales at the edge. The flesh is white, slightly thick or thick, fine and has a fragrance. When young, the edge is involute with white or yellowish-white fluff, which disappears as it grows. There is a velum under the pileus, which later ruptures to form an incomplete annulus. When mature, the pileus margin is involute and cracked; Lamellae: Radially arranged on the back of the pileus, white, soft, arranged in a blade shape, 1 - 4 mm wide; Stipe: Growing at the center or eccentrically under the pileus, cylindrical or nearly cylindrical, solid and tough, often curved, and connected to the mycelium in the substrate at the lower part. It is an organ that supports the pileus and transports nutrients and water, and its thickness and length are related to different nutrients, temperatures, light conditions and strains. Lentinula edodes in the world is mainly distributed in an arc-shaped area on the west side of the Pacific Ocean. It extends from Hokkaido in Japan in the north to Papua New Guinea in the south, and to the foothills of Dhaulagiri in Nepal in the west. In addition, there are Lentinula edodes varieties along the northern Mediterranean coast of Africa, similar Lentinula edodes are distributed in New Zealand, and it is also cultivated in Patagonia in South America. Lentinula edodes in China is mainly distributed in Anhui, Jiangsu, Shanghai, Zhejiang, Jiangxi, Hunan, Fujian, Taiwan, Guangdong, Guangxi, Yunnan, Guizhou, Sichuan and other places. Guangxi is known as "eight mountains, one water and one part field", naturally famous for its many mountains, and is rich in animal, plant and microbial resources. Among them, Lentinula edodes is distributed to varying degrees in the mixed forests of general alpine mountainous areas.

[0003] Lentinula edodes is the edible mushroom with the largest production in China. It has thick and tender flesh, delicious taste and unique aroma, and is rich in nutrients such as protein, amino acids, mineral elements and vitamins. It has the characteristics of high protein and low fat, and is a high-quality dietary protein source, and its protein quality is better than that of many fruits, vegetables and crops. Lentinula edodes also contains bioactive substances such as polysaccharides, and has the effects of reducing cholesterol, reducing blood pressure, antibacterial, antiviral, enhancing immunity, antioxidant, regulating intestinal microorganisms, antifungal, etc. It has extremely high nutritional, health care and medicinal values, and has been recommended by the Food and Agriculture Organization of the United Nations as a source of highly nutritious edible mushrooms and valuable medicinal compounds.

[0004] Therefore, finding new varieties of shiitake mushroom strains with more production application value is the current key research direction. Summary of the invention

[0005] In view of the shortcomings of the prior art, the present application provides a new shiitake mushroom strain (Yaoxiang No. 1), which has a sweeter and purer taste when cooked, a richer and more elegant aroma, and a refreshing aroma when dried. It is a shiitake mushroom strain with potential promotion and application value. The specific contents are as follows:

[0006] A new shiitake mushroom strain "Yaoxiang No. 1" (Lentinula edodes) was obtained by the inventor on January 23, 2022 from fruiting body samples collected from rotten wood of miscellaneous trees at a geographical location of 110°13′7.03″\24°9′38.11″\1225m in a mixed coniferous and broad-leaved forest in Changer Village, Jinxiu Town, Jinxiu County, Guangxi Province through tissue separation, purification, qualitative analysis and domestication. It is an excellent strain with production application value.

[0007] It has been deposited in the General Microbiology Center of China Culture Collection Administration (address: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences) on January 13, 2025, with the deposit number CGMCCNO.41776.

[0008] Furthermore, the new shiitake mushroom strain has a relatively thin and soft cap of its fruiting body, a relatively thin and long stipe, and relatively sparse spacing between gills.

[0009] Further, the new shiitake strain includes the spores, mycelium, and fruiting bodies of the strain.

[0010] The new mushroom strain described is used in food production and agricultural cultivation.

[0011] Further, the new shiitake strain includes the spores, mycelium, fruiting body, and extract of the new shiitake strain.

[0012] The Yaoxiang No. 1 shiitake mushroom strain of the present application is a shiitake mushroom strain with good mushroom production performance and excellent quality obtained through natural selection. It is about 120 days old and can produce mushrooms normally at a temperature of 15-28°C. It can obtain a bioconversion rate of more than 80% on the traditional mixed sawdust formula and the culture medium with mulberry branch chips accounting for 60%. Compared with the strains that are currently promoted and applied in the mass market through modern breeding methods such as hybridization, physical mutagenesis, and cell fusion, its fruiting body cap is thinner and softer, the stem is thinner and longer, and the distance between the gills is sparse ( Figures 24 - 27 ), but when cooked, the mushroom tastes sweeter and purer, the aroma is more rich, elegant and refreshing, and the smell after drying makes people feel refreshed. Therefore, Yaoxiang No. 1 fruiting body products are more suitable for the mid-to-high-end market.

[0013] The cultivation method of the new shiitake mushroom strain "Yaoxiang No. 1" is characterized in that the temperature range is controlled to be 5-35°C, and the cultivation is carried out using a traditional miscellaneous sawdust formula or a culture medium in which mulberry branch chips account for 60%.

[0014] Furthermore, the temperature range is 22-25°C.

[0015] Further, the cultivation materials used are: 78% wood chips, 20% bran, 1% gypsum, 1% lime powder, 60-65% moisture content, and pH 6.0-6.5.

[0016] Further, the cultivation materials used are: 60% mulberry crumbs, 20% miscellaneous wood chips, 18% bran, 1% gypsum, 1% lime powder, moisture content 60-65%, pH 6.0-6.5.

[0017] Beneficial effects of this application:

[0018] (1) The present application provides a new shiitake mushroom strain (Yaoxiang No. 1) (Lentinula edodes) that was newly isolated and successfully preserved. It was obtained by the inventor on January 23, 2022 from a fruiting body sample collected from rotten wood of miscellaneous trees at a geographical location of 110°13′7.03″\24°9′38.11″\1225m in a mixed coniferous and broad-leaved forest in Changer Village, Jinxiu Town, Jinxiu County, Guangxi Province through tissue separation, purification, characterization and domestication. It is an excellent strain with production application value.

[0019] (2) The new shiitake mushroom strain (Yaoxiang No. 1) provided in the present application is a shiitake mushroom strain with good mushroom-producing performance and excellent quality obtained through natural selection. It is about 120 days old and can produce mushrooms normally at a temperature of 15-28°C. It has a bioconversion rate of more than 80% on a traditional mixed sawdust formula and a culture medium with mulberry branch chips accounting for 60%. Compared with the strains that are currently promoted and applied in the mass market through modern breeding methods such as hybridization, physical mutagenesis, and cell fusion, its fruiting body cap is thinner and softer, its stipe is thinner and longer, and the distance between the gills is sparse ( Figures 24 - 27 ), the mushrooms taste sweeter and pure, the aroma is richer and more elegant, and refreshing. It makes people feel refreshed when drying and smelling. Therefore, Yaoxiang No. 1 fruit solid products are more suitable for mid-to-high-end markets. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figures 1 - 5 These are the fruiting body morphological characteristics of "Yaoxiang No. 1".

[0021] Figures 6 - 9 It is an artificial bag cultivation method for shiitake mushrooms.

[0022] Figures 10 - 12 It is a fruiting body morphological characteristics artificially cultivated by "Yaoxiang No. 1".

[0023] Figure 13 It is the culture characteristics of the mycelium on the plate.

[0024] Figure 14 It is the reddish-brown pigment secreted after the mycelium ages (plane photo).

[0025] Figure 15 It is the reddish-brown pigment secreted after the mycelium ages (photo against light).

[0026] Figure 16 It is the electrophoresis pattern (23 - Yaoxiang No.1; 24 - Lentinula edodes 135; 25 - Lentinula edodes 7401; 26 - Lentinula edodes 808; 27 - Qingyuan 20).

[0027] Figure 17 It is the identification result of Yaoxiang No.1 (J.C - 5), and it is a strain of Lentinula edodes.

[0028] Figure 18 It is the phylogenetic tree: J.C - 5 ~ Yaoxiang No.1; J.C - 6 ~ Lentinula edodes 135; J.C - 7 ~ Lentinula edodes 7401; J.C - 8 ~ Lentinula edodes 808; J.C - 9 ~ Qingyuan 20.

[0029] Figure 19 It is the antagonistic reaction between Yaoxiang No.1 and Lentinula edodes 135

[0030] Figure 20 It is the antagonistic reaction between Yaoxiang No.1 and Lentinula edodes 7401.

[0031] Figure 21 It is the antagonistic reaction between Yaoxiang No.1 and Lentinula edodes 808.

[0032] Figure 22 It is the antagonistic reaction between Yaoxiang No.1 and Qingyuan 20.

[0033] Figure 23 It is the antagonistic reaction between Yaoxiang No.1 and Wuxiang No.1.

[0034] Figure 24 It is the appearance morphological characteristics of the fruiting body of Lentinula edodes 808.

[0035] Figure 25 It is the appearance morphological characteristics of the fruiting body of Yaoxiang No.1.

[0036] Figure 26 It is the characteristics of the pileus, stipe and lamella of the fruiting body of Lentinula edodes 808.

[0037] Figure 27 It is the characteristics of the pileus, stipe and lamella of the fruiting body of Yaoxiang No.1. Specific implementation manners

[0038] The following further elaborates on the present application in combination with specific embodiments, which is an explanation rather than a limitation of the present application.

[0039] The present invention relates to a new Lentinula edodes strain "Yaoxiang No. 1", belonging to the technical field of edible and medicinal fungi.

[0040] 1. Relevant information of the strain: including functions, classification, properties, etc.

[0041] ① Taxonomic status: Lentinula edodes belongs to the kingdom Eumycetes, phylum Eumycota (Mycobionta), class Basidiomycetes, order Agaricales, family Omphalotaceae, and genus Lentinus.

[0042] ② Characteristic features (including fruiting body morphological features, mycelium culture features, biological characteristics, cultivation characteristics, etc.):

[0043] a. Fruiting body morphological features: Lentinula edodes is also known as fragrant mushroom, shiitake mushroom, flower mushroom, scented mushroom, fragrant letter, shiitake, winter mushroom, thick mushroom, etc.; wild Lentinula edodes fruiting bodies are mostly solitary, and the fruiting bodies of Lentinula edodes cultivated by artificial bag cultivation can be solitary, clustered, or growing in groups, with the fruiting bodies medium to slightly large. The mature fruiting body is umbrella-shaped and consists of a pileus, gills, and stipe. Pileus: Located at the top of the stipe, circular, with a diameter generally of 3 - 15 cm, and up to 20 cm for large ones. It is hemispherical when young and then becomes flat to slightly flat. The surface color ranges from chestnut color, light brown, dark brown to dark cinnamon color, and there are often dark scales in the middle, while there are often dirty white hairy or flocculent scales at the edge. The flesh is white, slightly thick or thick, fine, and has a fragrance. The edge is involute when young and has white or yellowish-white villi, which disappear as it grows. There is a velum under the pileus, which later breaks and forms an incomplete annulus. The pileus margin is reflexed and cracked when old; Gills: Radially arranged on the back of the pileus, white, soft, arranged in a blade shape, 1 - 4 mm wide; Stipe: Growing in the center or eccentrically under the pileus, cylindrical or nearly cylindrical, solid and tough, often curved, with the lower part connected to the mycelium in the substrate, and it is an organ that supports the pileus and transports nutrients and water. Its thickness and length are related to different nutrients, temperatures, light conditions, and strains. Lentinula edodes in the world is mainly distributed in an arc-shaped area on the west side of the Pacific Ocean. It extends from Hokkaido in Japan in the north to Papua New Guinea in the south and to the foothills of Dhaulagiri in Nepal in the west. In addition, there are Lentinula edodes variants along the northern Mediterranean coast of Africa, similar Lentinula edodes are distributed in New Zealand, and it is also cultivated in Patagonia in South America. Lentinula edodes in China is mainly distributed in Anhui, Jiangsu, Shanghai, Zhejiang, Jiangxi, Hunan, Fujian, Taiwan, Guangdong, Guangxi, Yunnan, Guizhou, Sichuan and other places. Guangxi is known as "eight mountains, one water and one field", naturally famous for its many mountains, and is very rich in animal, plant, and microbial resources. Among them, Lentinula edodes is distributed to varying degrees in the mixed forests of general alpine mountains.

[0044] The fruiting body morphological features of "Yaoxiang No. 1" are asFigures 1 - 5 , the artificial bag cultivation technology of Lentinula edodes is relatively mature (such as Figures 6 - 9 ), Yaoxiang No. 1 is a superior strain with production application value obtained by tissue separation, purification, qualitative determination, and domestication from the fruiting body samples collected from the miscellaneous tree rotten wood at the geographical location of 110°13′7.03″\24°9′38.11″\1225m in the broad-leaved and coniferous mixed forest in the southwest direction of Chang'er Village, Jinxiu Town, Jinxiu County, Guangxi. The morphological characteristics of its artificially cultivated fruiting bodies are similar to those of wild ones (such as Figures 10 - 12 ).

[0045] b. Mycelium culture characteristics The mycelium of Yaoxiang No. 1 on the conventional comprehensive potato medium [Potato (peeled) 200g, glucose 20g, potassium dihydrogen phosphate 3.0g, magnesium sulfate 1.5g, vitamin B1 10mg, water 1000ml, pH natural.] has the following culture characteristics: The mycelium is white, dense, round, thick and strong, with short aerial hyphae. The substrate mycelium is relatively thick. As the culture time prolongs, the mycelium will gradually age and secrete reddish-brown pigments in a certain area of the colony (below Figures 13 - 15 ); Its mycelium can grow within the temperature range of 5 - 35°C, and the suitable growth temperature range is 22 - 25°C. However, the mycelium growth is very poor below 10°C and above 30°C, and it will die when the temperature exceeds 40°C.

[0046] Yaoxiang No. 1 is identified as a variety of Lentinula edodes by BGI ( Figures 16 - 18 );

[0047] When Yaoxiang No. 1 is compared with the production strains Lentinula edodes 135, Lentinula edodes 7401, Lentinula edodes 808, and Qingyuan 20, there are significant antagonistic reactions [Note: Individuals with significantly different genetic backgrounds will form obvious dividing lines in the junction area, which is the antagonistic reaction. The antagonistic reaction is a specific manifestation of somatic non-affinity and has been applied in the preliminary study of fungal genetic relationships due to its advantages of rapidity, convenience, and intuitiveness. It was once the main means for fungal strain identification.]. ( Figure 19 , 20 ).

[0048] c. Cultivation: Yao Xiang No.1 is a Lentinula edodes strain obtained through natural selection, with excellent mushroom-producing performance and quality. The suitable temperature for mycelial growth is 22 - 25°C, and the incubation time is about 120 days. It can normally form mushrooms within the temperature range of 15 - 28°C, and a biological conversion rate of over 80% can be achieved on both the traditional sawdust formula and the culture medium with 60% mulberry branch sawdust. Its artificial bag cultivation procedure is similar to the conventional Lentinula edodes cultivation technique, and it goes through processes such as making spawn bags, mycelial cultivation, spawn bag color transformation, mushroom management, harvesting, and tide turning. A. Cultivation medium formula: ① 78% sawdust, 20% wheat bran, 1% gypsum, 1% lime powder, water content 60 - 65%, pH 6.0 - 6.5; ② 60% mulberry branch sawdust, 20% sawdust, 18% wheat bran, 1% gypsum, 1% lime powder, water content 60 - 65%, pH 6.0 - 6.5. B. Brief introduction to cultivation techniques --- ① Spawn bag making: After the raw materials are fully stirred evenly, they are packed into 15 cm × 55 cm × 0.05 mm polypropylene spawn bags and the bag mouths are tied; ② Sterilization: After being sent into an atmospheric or high-pressure sterilizer for thorough sterilization according to the established sterilization parameters, the spawn bags are removed from the sterilizer and sent to a pre-cooling room for forced cooling or natural cooling; ③ Inoculation: When the temperature of the spawn bags cools below 30°C, start inoculating the spawn bags according to the aseptic operation procedure requirements and an inoculation plan of 3 - 4 holes per bag; ④ Mycelial cultivation: After inoculation, immediately send them to the indoor shelf at 22 - 25°C for mycelial cultivation (i.e., "incubation"); After-ripening and color transformation: After about 120 days, the mycelial cultivation is approaching maturity, and the spawn bags need to be punctured to promote after-ripening and color transformation, and this process takes about 15 days; ⑤ Startling the mycelium and removing the bags for mushroom growth: When the spawn bags have basically turned yellow-brown and there are tubercles, first perform the startling mycelium treatment, that is, hold one bag in each hand and strike them against each other, then remove the plastic bags, and place the bagless spawn bags on the shelf for mushroom management --- If the spawn bags are not severely dehydrated, there is no need to replenish water, on the contrary, water needs to be replenished. The first flush of mushrooms can appear as mushroom buds under the condition of a temperature difference of 5 - 8°C about 5 days after placing the spawn bags, and the first flush of mushrooms can be harvested after about 15 days; ⑥ Rehydrating, startling the mycelium, and turning the tide: After harvesting the first flush of mushrooms, let the spawn bags rest for about 15 days, then startle the mycelium and rehydrate to induce the second flush of mushrooms. The treatment and management methods for the third and fourth flushes of mushrooms are the same, and a total of 4 - 5 flushes of mushrooms can be harvested, and the mushroom-growing period lasts about 5 months.

[0049] 2. Preservation information of the strain: Preserved in the China General Microbiological Culture Collection Center (Address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences) on January 13, 2025, with the preservation number CGMCC NO. 41776

[0050] 3. Ways to obtain the strain: On January 23, 2022, the inventor collected fruiting body samples from rotten wood of miscellaneous trees at a geographical location of 110°13′7.03″\24°9′38.11″\1225m in a mixed coniferous and broad-leaved forest in the southwest of Changer Village, Jinxiu Town, Jinxiu County, Guangxi, and obtained an excellent strain with production application value through tissue separation, purification, qualitative analysis and domestication.

[0051] 4. Characteristics of Yaoxiang No. 1 strain

[0052] Yaoxiang No. 1 is a shiitake mushroom strain with good mushroom production performance and excellent quality obtained through natural selection. It is about 120 days old and can produce mushrooms normally at a temperature of 15-28℃. It has a bioconversion rate of more than 80% on the traditional mixed sawdust formula and the culture medium with mulberry branch chips accounting for 60%. Compared with the strains currently promoted and applied in the mass market through modern breeding methods such as hybridization, physical mutagenesis, and cell fusion, its fruiting body cap is thinner and softer, the stem is thinner and longer, and the distance between the gills is sparser (the comparison data is shown in the following table) ( Figures 24 - 27 ), the inventors have repeatedly used the fruiting bodies of Shiitake 135 and Shiitake 808 as controls, and have fully proved through actual cooking and drying comparison tests at the same level, as well as actual evaluations by consumers, that the fruiting bodies of Yaoxiang No. 1 fill the house with fragrance when cooking, and its Shiitake flavor is sweeter and purer, its aroma is richer and more elegant, and refreshing. After drying, the fragrance is more refreshing, and it is deeply favored by consumers. Therefore, the fruiting body products of Yaoxiang No. 1 are more suitable for the mid-to-high-end market in the form of gift boxes.

[0053]

[0054] 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 new Lentinula edodes strain, characterized in that, It is preserved in the General Microbiological Center of the China Committee for Culture Collection of Microorganisms, with the preservation number of CGMCC NO. 41776.

2. The new Lentinula edodes strain according to claim 1, characterized in that, For the said new Lentinula edodes strain, the pileus of its fruiting body is relatively thin and soft, the stipe is relatively thin and long, and the distance between the gills is relatively sparse.

3. The new shiitake mushroom strain according to claim 1, characterized in that, The said new Lentinula edodes strain includes the spores, mycelia, and fruiting bodies of this strain.

4. Application of the new Lentinula edodes strain described in Claim 1 in food production.

5. Application of the new Lentinula edodes strain described in Claim 1 in agricultural cultivation.

6. The application according to claims 4 and 5, characterized in that, The said new Lentinula edodes strain includes the spores, mycelia, fruiting bodies, and extracts of this new Lentinula edodes strain.

7. The cultivation method of the new Lentinula edodes strain according to claim 1, characterized in that, The controlled temperature range is 5 - 35°C.

8. The cultivation method of the new Lentinula edodes strain according to claim 7, characterized in that, The said temperature range is 22 - 25°C.

9. The cultivation method of the new Lentinula edodes strain according to claim 7, characterized in that, The cultivation medium used is: 78% miscellaneous sawdust, 20% wheat bran, 1% gypsum, 1% lime powder, moisture content 60 - 65%, pH 6.0 - 6.

5.

10. The cultivation method of the new Lentinula edodes strain according to claim 7, characterized in that, The cultivation medium used is: 60% mulberry branch sawdust, 20% miscellaneous sawdust, 18% wheat bran, 1% gypsum, 1% lime powder, moisture content 60 - 65%, pH 6.0 - 6.5.

Images